Effects of dexamethasone administration on serum trypsin-like immunoreactivity in healthy dogs.
To determine whether administration of dexamethasone altered serum trypsin-like immunoreactivity (TLI) in healthy dogs. 12 healthy dogs. Dexamethasone (0.25 mg/kg, p.o., q 24 h) was administered for 7 days. Serum TLI, alpha-amylase and alanine aminotransferase (ALT) activities, and urea and creatinine concentrations were determined on days 0, 7, 14, and 21 of the study. Serum TLI and ALT activities were significantly increased, and serum alpha-amylase activity was significantly decreased after administration of dexamethasone for 7 days. However, values obtained on days 14 and 21 were not significantly different from baseline values. Dexamethasone administration was not associated with any significant changes in serum creatinine or urea concentrations. Serum TLI and alpha-amylase activities were significantly correlated prior to dexamethasone administration. Dogs did not develop clinical signs of pancreatitis. Dexamethasone administration was associated with an increase in serum TLI. However, values returned to baseline 7 days after dexamethasone administration was discontinued. Serum TLI may be falsely high in dogs that have been treated with dexamethasone in the week preceding analysis.


Magnetic molecularly imprinted polymers for recognition and enrichment of polysaccharides from seaweed.
New magnetic molecularly imprinted polymers with two templates were fabricated for the recognition of polysaccharides (fucoidan and alginic acid) from seaweed by magnetic solid-phase extraction, and the materials were modified by seven types of deep eutectic solvents. It was found that the deep eutectic solvents magnetic molecularly imprinted polymers showed stronger recognition and higher recoveries for fucoidan and alginic acid than magnetic molecularly imprinted polymers, and the deep eutectic solvents-4-magnetic molecularly imprinted polymers had the best effects. The practical recovery of the two polysaccharides (fucoidan and alginic acid) purified with deep eutectic solvents-4-magnetic molecular imprinted polymers in seaweed under the optimal conditions were 89.87, and 92.0%, respectively, and the actual amounts extracted were 20.6 and 18.7 μg/g, respectively. To sum up, the developed method proved to be a novel and promising method for the recognition of complex polysaccharide samples from seaweed.


[Total communication in aphasia therapy].
Total Communication Therapy is based on the possibilities which aphasic patients have left to communicate. All aspects of language are trained. The environment stimulates the patient and uses all possible means of communication in order to achieve an optimal level of communication. An evaluation test is being prepared at the moment.


Practical modeling of Bayesian decision problems - exploiting deterministic relations.
The widespread use of influence diagrams to represent and solve Bayesian decision problems is still limited by the inflexibility and rather restrictive semantics of influence diagrams. We propose a number of extensions and adjustments to the definition of influence diagrams in order to make the practical use of influence diagrams more flexible and less restrictive. In particular, we describe how deterministic relations can be exploited to increase the flexibility and efficiency of representing and solving Bayesian decision problems. The issues addressed in the paper were motivated by the construction of a decision support system for mission management of unmanned underwater vehicles (UUVs).


Responsiveness and interpretability of the Animated Activity Questionnaire for assessing activity limitations of patients with hip or knee osteoarthritis.
The aim of the study was to determine the responsiveness and interpretability of the Animated Activity Questionnaire (AAQ), an online questionnaire in which osteoarthritis patients select animations that best match their performance of daily activities. A longitudinal study was carried out, in which 94 patients with hip or knee osteoarthritis were assessed at baseline, and 3 and 6 months after treatment (conservative and surgical). Responsiveness was assessed by means of testing hypotheses about expected correlations between change in AAQ, a Global Rating Scale of change (GRS) and change in the Activities of Daily Living subscale of the Hip disability or Knee injury and Osteoarthritis Outcome Score (H/KOOS), and a combination of performance-based tests (the 30 s chair-stand test, the timed up-and-go test and the nine-step stair climbing test). The minimal important change (MIC) was estimated by means of the receiving operating characteristics (ROC) method. The correlations of the AAQ with the H/KOOS were as expected, but other correlations were lower than anticipated. The area under the ROC curve was 0.74 at 6 months. At 3 months' follow-up, the correlations were too low to calculate a MIC. A total of 20% of the results at 3 months and 80% of the results at 6 months were in accordance with the hypotheses. The MIC was 9 points at 6 months. The AAQ was sufficiently responsive at the six-months follow-up, but not at the three-month follow-up. The MIC at the 6-month follow-up (9 points) was slightly lower than the smallest detectable change of 14 points found in a previous study.


Lessons learned in developing a culturally adapted intervention for African-American families coping with parental cancer.
Prior clinical research supports the effectiveness of cancer support groups for cancer patients and their families, yet African-American families continue to be underrepresented in cancer support groups and in cancer clinical research studies. In order to fill this gap, we developed and evaluated a culturally adapted family support group for African-American families coping with parental cancer. We encountered unexpected challenges in overcoming barriers to recruitment, partnering with oncology providers, and building trust with the African-American community and African-American families coping with parental cancer. We describe actions taken during the two phases of this study and lessons learned along the way about recruiting and engaging African-American families in cancer support group studies, partnering with oncology providers, networking with the African-American community, and the importance of demonstrating cultural sensitivity to overcome the understandable historical legacy of mistrust.


A method for counting PCR template molecules with application to next-generation sequencing.
Amplification by polymerase chain reaction is often used in the preparation of template DNA molecules for next-generation sequencing. Amplification increases the number of available molecules for sequencing but changes the representation of the template molecules in the amplified product and introduces random errors. Such changes in representation hinder applications requiring accurate quantification of template molecules, such as allele calling or estimation of microbial diversity. We present a simple method to count the number of template molecules using degenerate bases and show that it improves genotyping accuracy and removes noise from PCR amplification. This method can be easily added to existing DNA library preparation techniques and can improve the accuracy of variant calling.


Prevalence and determinants of complementary and alternative medicine use during pregnancy: results from a nationally representative sample of Australian pregnant women.
Pregnant women have been identified as high users of complementary and alternative medicine (CAM). However, no research to date has provided a detailed analysis of the prevalence and determinants of CAM consumption amongst pregnant women. To examine the prevalence and determinants of CAM use by pregnant women, utilising a national representative sample. The study sample was obtained via the Australian Longitudinal Study on Women's Health. This paper is based on a sub-study of 1,835 pregnant women, administered in 2010. The women answered questions about CAM use, demographics, pregnancy-related health concerns and health service utilisation. Complementary and alternative medicine use was found to be high with 48.1% (n = 623) of pregnant women consulting a CAM practitioner and 52.0% (n = 842) of women using CAM products (excluding vitamins and minerals) during pregnancy. CAM practitioner visits were more likely for selected pregnancy-related health concerns, namely back pain or back ache, neck pain and labour preparation. Women were less likely to consult a CAM practitioner if they suffered with headaches/migraines. Employment was also found to be predictive of pregnant women's visits to a CAM practitioner. Significant health history and demographic predictors of CAM product use were tiredness and fatigue, embarking on preparation for labour and having a university education. Most pregnant women are utilising CAM products and/or services as part of their maternity care and obstetricians, general practitioners and midwives need to enquire with women in their care about possible CAM use to help promote safe, effective coordinated maternity care.


From Structure to Function: pH-Switchable Antimicrobial Nano-Self-Assemblies.
Stimuli-responsive nanocarriers based on lipid self-assemblies have the potential to provide targeted delivery of antimicrobial peptides, limiting their side effects while protecting them from degradation in the biological environments. In the present study, we design and characterize a simple pH-responsive antimicrobial nanomaterial, formed through the self-assembly of oleic acid (OA) with the human cathelicidin LL-37 as a model for an amphiphilic antimicrobial peptide. Colloidal transformations from core-shell cylindrical micelles with a cross-sectional diameter of ∼5.5 nm and a length of ∼23 nm at pH 7.0 to aggregates of branched threadlike micelles at pH 5.0 were detected using synchrotron small-angle X-ray scattering, cryogenic transmission electron microscopy, and dynamic light scattering. Biological in vitro assays using an Escherichia coli bacteria strain showed high antimicrobial activity of the positively charged LL-37/OA aggregates at pH 5.0, which was not caused by the pH conditions themselves. Contrary to that, negligible antimicrobial activity was observed at pH 7.0 for the negatively charged cylindrical micelles. The nanocarrier's ability to switch its biological activity "on" and "off" in response to changes in pH could be used to focus the antimicrobial peptides' action to areas of specific pH in the body. The presented findings contribute to the fundamental understanding of lipid-peptide self-assembly and may open up a promising strategy for designing simple pH-responsive delivery systems for antimicrobial peptides.


Health-related quality of life of fracture victims four years after the 2008 Sichuan earthquake.
To investigate health-related quality of life and its relationship with functional status and other related factors in victims with fractures 4 years after the 2008 Sichuan earthquake. A cross-sectional survey with a multi-stage random sampling method. Five hospitals from the areas most severely affected by the 2008 Sichuan earthquake. Victims with fractures aged 14 years and older who were hospitalized in the rehabilitation departments of the 5 identified hospitals during the period 12 May 2008 to 12 May 2009. Information on demographics, such as age, gender, marital status and educational level, functional status, working status, income, and health-related quality of life, were investigated. Manual muscle test, visual analogue scale, Modified Barthel Index and Medical Outcomes Short Form 36 (SF-36) were employed as the main outcome measures. A total of 243 victims with fractures were interviewed. Thirty-seven percent of the fracture victims had decreased muscle strength, 28.8% had limited range of motion, 51.8% still experienced pain, and 17.7% were dependent to different extents. With the exception of the domains vitality and mental health, the earthquake victims perceived significantly lower health-related quality of life than the local general population. Older age, being female, unmarried, low education, multiple fractures, muscle weakness, pain and being dependent were significant predictors of lower health-related quality of life. Most SF-36 subscales were negatively correlated with age, multiple fractures and pain, but positively correlated with independence in activities of daily living and income. Four years after the major Sichuan earthquake, many victims with fractures still had reduced functional status and experienced pain, and their health-related quality of life was low compared with the general population.


Effects and interactions of opioids on plasma exudation induced by cigarette smoke in guinea pig bronchi.
The effects of opioids on cigarette smoke-induced plasma exudation were investigated in vivo in the main bronchi of anesthetized guinea pigs, with Evans blue dye as a plasma marker. Acute inhalation of cigarette smoke increased plasma exudation by 216% above air control values. Morphine, 0.1-10 mg/kg but not 30 mg/kg, inhibited the exudation but had no significant effect on substance P-induced exudation. Both 10 and 30 mg/kg of morphine increased exudation in air control animals, an effect inhibited by antihistamines but not by a tachykinin neurokinin type 1-receptor antagonist. Naloxone inhibited all morphine responses. Cigarette smoke-induced plasma exudation was inhibited by a mu-opioid-receptor agonist (DAMGO) but not by agonists at delta (DPDPE)- or kappa (U-50488H)-receptors. None of these agonists affected exudation in air control animals. DPDPE prevented the inhibition by DAMGO of cigarette smoke-induced plasma exudation, and the combination of DAMGO and DPDPE increased exudation in air control animals. Prevention of inhibition and the combination-induced increase were inhibited by antihistamines or the mast cell-stabilizing drug sodium cromoglycate. U-50488H did not alter the response to either DAMGO or DPDPE. We conclude that, in guinea pig main bronchi in vivo, mu-opioid-receptor agonists inhibit cigarette smoke-induced plasma exudation via a prejunctional mechanism. Plasma exudation induced by mu- and delta-receptor interactions is due to endogenous histamine release from mast cells.


Oxovanadium(IV) complexes of curcumin for cellular imaging and mitochondria targeted photocytotoxicity.
Oxovanadium(IV) complexes [VO(R-tpy)(cur)](ClO4) (1, 2) of curcumin (Hcur) and terpyridine ligands (R-tpy) where R is phenyl (phtpy in 1) or p-triphenylphosphonium methylphenyl bromide (C6H4CH2PPh3Br) (TPP-phtpy in 2) were prepared and characterized and their DNA photocleavage activity, photocytotoxicity and cellular localization in cancer cells (HeLa and MCF-7) were studied. Acetylacetonate (acac) complexes [VO(R-tpy)(acac)](ClO4) of phtpy (3) and TPP-phtpy (4) were prepared and used as the control species. These complexes showed efficient cleavage of pUC19 DNA in visible light of 454 nm and near-IR light of 705 nm. Complexes 1 and 2 showed significant photocytotoxicity in visible light of 400-700 nm. FACS analysis showed sub-G1/G0 phase cell-cycle arrest in cancer cells when treated with 1 and 2 in visible light in comparison with the dark controls. Fluorescence microscopic studies revealed specific localization of the p-triphenylphosphonium complex 2 in the mitochondria of MCF-7 cancer cells whereas no such specificity was observed for complex 1.


Botulism mimicking Guillain-Barre syndrome: The question of plasma exchange in an unusual case of acute paralysis.
Guillain-Barré syndrome (GBS) is an immune-mediated polyradiculoneuropathy and the most common cause of acute flaccid paralysis worldwide. GBS classically presents with acute, progressive, ascending weakness, reduced to absent reflexes, and albuminocytological dissociation on cerebrospinal fluid (CSF) analysis. Botulism is a neurotoxin-mediated acute descending flaccid paralysis with cranial nerve palsies and dysautonomia. Botulism in adults is caused by ingestion/inhalation of botulinum toxin or wound infection with Clostridium botulinum. Both GBS and botulism can rapidly precipitate respiratory failure; thus, prompt diagnosis and treatment are crucial to mitigate poor outcomes. Herein, we describe a case of botulism initially diagnosed as GBS given classic laboratory features, and describe the importance of careful consideration of the most appropriate therapeutic modalities in cases of acute flaccid paralysis, particularly regarding empiric administration of botulinum antitoxin and use of intravenous immune globulin in lieu of plasma exchange for potential GBS to prevent removal of antitoxin.


Synthetic lethal interaction of cetuximab with MEK1/2 inhibition in NRAS-mutant metastatic colorectal cancer.
KRAS mutations are an established predictor of lack of response to EGFR-targeted therapies in patients with metastatic colorectal cancer (mCRC). However, little is known about the role of the rarer NRAS mutations as a mechanism of primary resistance to the anti-EGFR monoclonal antibody cetuximab in wild-type KRAS mCRC. Using isogenic mCRC cells with a heterozygous knock-in of the NRAS activating mutation Q61K, we aimed to elucidate the mechanism(s) by which mutant NRAS blocks cetuximab from inhibiting mCRC growth. NRASQ61K/+ cells were refractory to cetuximab-induced growth inhibition. Pathway-oriented proteome profiling revealed that cetuximab-unresponsive ERK1/2 phosphorylation was the sole biomarker distinguishing cetuximab-refractory NRASQ61K/+ from cetuximab-sensitive NRAS+/+ cells. We therefore employed four representative MEK1/2 inhibitors (binimetinib, trametinib, selumetinib, and pimasertib) to evaluate the therapeutic value of MEK/ERK signaling in cetuximab-refractory NRAS mutation-induced mCRC. Co-treatment with an ineffective dose of cetuximab augmented, up to more than 1,300-fold, the cytotoxic effects of pimasertib against NRASQ61K/+ cells. Simultaneous combination of MEK1/2 inhibitors with cetuximab resulted in extremely high and dose-dependent synthetic lethal effects, which were executed, at least in part, by exacerbated apoptotic cell death. Dynamic monitoring of real-time cell growth rates confirmed that cetuximab synergistically sensitized NRASQ61K/+ cellsto MEK1/2 inhibition. Our discovery of a synthetic lethal interaction of cetuximab in combination with MEK1/2 inhibition for the NRAS mutant subgroup of mCRC underscores the importance of therapeutic intervention both in the MEK-ERK and EGFR pathways to achieve maximal therapeutic efficacy against NRAS-mutant mCRC tumors.


[Genetic determination of dyslipidemia - What tell us the results of genome-wide association studies?]
Dyslipidemia (high levels of plasma triglycerides and total cholesterol/LDL-cholesterol and low HDL-cholesterol) is considered as one of the major factors in the development of atherosclerosis and subsequent myocardial infarction. The final value of lipid parameters results from joint action of genetic predispositions and lifestyle factors (primarily smoking status, physical activity and in lower extent also diet). It is estimated that genetic factors are responsible for 40-80 % of the variability of plasma lipid values. Currently are as predictors DL analyzed mainly single nucleotide polymorphisms (SNPs). A fundamental shift in knowledge of genetic determination DL bring genome-wide association studies (GWAs). These revealed several dozen major polymorphisms in a DNA sequence related to lipid levels. Rather surprisingly, these variants are usually not substitutions of the amino acids, or causing a premature stop codon, but substitutions outside the genes. GWAS also found a number of variants within the genes whose function in lipid metabolism was completely unknown (e.g. gene for sortilin). Polymorphisms in genes for APOE, SORT1, LDLR (affect levels of total cholesterol and LDL-cholesterol), CETP, APOA1, ABCA-1, GALNT-2 (influence HDL-cholesterol) and finally in genes for APOA5, LPL or TRIB1 (affect the levels of triglycerides) but explains max. 30 % of the variability of plasma lipids. It is supposed, that rare polymorphisms/mutations and genetic factors unrelated directly to alterations in the DNA sequence (DNA methylation, histone modifications, regulatory RNA molecules) are responsible for the remaining proportion of DL determination.Key words: gene - cholesterol - interaction - mutation - polymorphism - triglycerides.


Particulate Matter Air Pollution is a Significant Risk Factor for Cardiovascular Disease.
Air pollution is responsible worldwide for 9-12 million deaths annually. The major contributor to air pollution is particulate matter ≤2.5 µg per cubic meter of air (PM2.5) from vehicles, industrial emissions, and wildfire smoke. United States ambient air standards recommend annual average PM2.5 concentrations of ≤12 μg/m³ while European standards allow an average annual PM2.5 concentration of ≤20 μg/m3. However, significant PM2.5 cardiovascular and pulmonary health risks exist below these concentrations. Chronic PM2.5 exposure significantly increases major cardiovascular and pulmonary event risks in Americans by 8 to more than 20% for each 10-μg/m3 increase in PM2.5. PM2.5-induced increases in lipid peroxidation, induction of vascular inflammation and endothelial cell injury initiate and propagate respiratory diseases, coronary and carotid atherosclerosis. PM2.5 can cause atherosclerotic vascular plaque rupture and myocardial infarction and stroke by activating metalloproteinases. This article discusses PM2.5 effects on the cardiovascular and pulmonary systems, specific PM2.5 pathophysiologic mechanisms contributing to cardiopulmonary disease, and preventive measures to limit the cardiovascular and pulmonary effects of PM2.5.


Generalization of fear inhibition by disrupting hippocampal protein synthesis-dependent reconsolidation process.
Repetitive replay of fear memories may precipitate the occurrence of post-traumatic stress disorder and other anxiety disorders. Hence, the suppression of fear memory retrieval may help prevent and treat these disorders. The formation of fear memories is often linked to multiple environmental cues and these interconnected cues may act as reminders for the recall of traumatic experiences. However, as a convenience, a simple paradigm of one cue pairing with the aversive stimulus is usually used in studies of fear conditioning in animals. Here, we built a more complex fear conditioning model by presenting several environmental stimuli during fear conditioning and characterize the effectiveness of extinction training and the disruption of reconsolidation process on the expression of learned fear responses. We demonstrate that extinction training with a single-paired cue resulted in cue-specific attenuation of fear responses but responses to other cures were unchanged. The cue-specific nature of the extinction persisted despite training sessions combined with D-cycloserine treatment reveals a significant weakness in extinction-based treatment. In contrast, the inhibition of the dorsal hippocampus (DH) but not the basolateral amygdala (BLA)-dependent memory reconsolidation process using either protein synthesis inhibitors or genetic disruption of cAMP-response-element-binding protein-mediated transcription comprehensively disrupted the learned connections between fear responses and all paired environmental cues. These findings emphasize the distinct role of the DH and the BLA in the reconsolidation process of fear memories and further indicate that the disruption of memory reconsolidation process in the DH may result in generalization of fear inhibition.


Control strategies to re-establish glenohumeral stability after shoulder injury.
Muscles are important "sensors of the joint instability". The aim of this study was to identify the neuro-motor control strategies adopted by patients with anterior shoulder instability during overhead shoulder elevation in two planes. The onset, time of peak activation, and peak magnitude of seven shoulder muscles (posterior deltoid, bilateral upper trapezius, biceps brachii, infraspinatus, supraspinatus and teres major) were identified using electromyography as 19 pre-operative patients with anterior shoulder instability (mean 27.95 years, SD = 7.796) and 25 age-matched asymptomatic control subjects (mean 23.07 years, SD = 2.952) elevated their arm above 90 degrees in the sagittal and coronal planes. Temporal characteristics of time of muscle onsets were significantly different between groups expect for teres major in the coronal plane (t = 1.1220, p = 0.2646) Patients recruited the rotator cuff muscles earlier and delayed the onset of ipsilateral upper trapezius compared with subjects (p<0.001) that control subjects. Furthermore, significant alliances existed between the onsets of infraspinatus and supraspinatus (sagittal: r = 0.720; coronal: r = 0.756 at p<0.001) and ipsilateral upper trapezius and infraspinatus (sagittal: r = -0.760, coronal: r = -0.818 at p<0.001). The peak activation of all seven muscles occurred in the mid-range of elevation among patients with anterior shoulder instability whereas subjects spread peak activation of all 7 muscles throughout range. Peak magnitude of patients' infraspinatus muscle was six times higher (sagittal: t = -8.6428, coronal: t = -54.1578 at p<0.001) but magnitude of their supraspinatus was lower (sagittal: t = 36.2507, coronal: t = 35.9350 at p<0.001) that subjects. Patients with anterior shoulder instability adopted a "stability before mobility" neuro-motor control strategy to initiate elevation and a "stability at all cost" strategy to ensure concavity compression in the mid-to-150 degrees of elevation in both sagittal and coronal planes.


Simultaneous glucocorticoid and noradrenergic activity disrupts the neural basis of goal-directed action in the human brain.
Stress promotes a shift from goal-directed action-outcome learning toward habitual stimulus-response learning. This shift is mediated by an interaction of noradrenergic activity and glucocorticoid stress hormones. In the present experiment, we examined the neural correlates of the stress (hormone)-induced shift from goal-directed to habit learning in the human brain. Healthy participants were administered hydrocortisone, the α2-adrenoceptor antagonist yohimbine, or both before they were trained in two instrumental actions leading to two distinct food rewards. After training, one of the rewards was devalued by feeding participants to satiety on that food. Finally, participants were presented the two instrumental actions in extinction. We collected functional magnetic resonance images both during instrumental training and during extinction testing. Our behavioral data confirmed that the simultaneous administration of hydrocortisone and yohimbine renders instrumental behavior insensitive to the outcome devaluation (and thus habitual), whereas hydrocortisone or yohimbine alone have no such effect. At the neural level, the combined administration of hydrocortisone and yohimbine reduced the sensitivity of the orbitofrontal and medial prefrontal cortex to changes in outcome value. Brain areas that have been previously implicated in habit learning were not modulated by hydrocortisone and yohimbine. These findings suggest that concurrent glucocorticoid and noradrenergic activity disrupts the neural bases of goal-directed action and thus renders behavior habitual.


Fanconi anemia complementation group A (FANCA) protein has intrinsic affinity for nucleic acids with preference for single-stranded forms.
The Fanconi anemia complementation group A (FANCA) gene is one of 15 disease-causing genes and has been found to be mutated in ∼60% of Fanconi anemia patients. Using purified protein, we report that human FANCA has intrinsic affinity for nucleic acids. FANCA binds to both single-stranded (ssDNA) and double-stranded (dsDNA) DNAs; however, its affinity for ssDNA is significantly higher than for dsDNA in an electrophoretic mobility shift assay. FANCA also binds to RNA with an intriguingly higher affinity than its DNA counterpart. FANCA requires a certain length of nucleic acids for optimal binding. Using DNA and RNA ladders, we determined that the minimum number of nucleotides required for FANCA recognition is ∼30 for both DNA and RNA. By testing the affinity between FANCA and a variety of DNA structures, we found that a 5'-flap or 5'-tail on DNA facilitates its interaction with FANCA. A patient-derived FANCA truncation mutant (Q772X) has diminished affinity for both DNA and RNA. In contrast, the complementing C-terminal fragment of Q772X, C772-1455, retains the differentiated nucleic acid-binding activity (RNA > ssDNA > dsDNA), indicating that the nucleic acid-binding domain of FANCA is located primarily at its C terminus, where most disease-causing mutations are found.


Flexural versus plaque lesions in psoriasis: an immunohistochemical differentiation.
Clinical presentation, therapeutic options, micro-environment and HLA-typing have been reported to be different in flexural psoriasis as compared to plaque psoriasis. We were interested in any difference concerning the pathogenesis of both conditions. By analysing T-cell subsets, NK-T cells and proliferation and differentiation markers, insight into the pathogenesis of both subtypes was obtained. Quantitative studies of T-cell subsets, cells expressing NK-receptors and markers of proliferation and differentiation in flexural and plaque psoriasis were investigated. Biopsies from 6 patients with both flexural and plaque lesions were obtained and processed for immunohistochemistry. Several T-cell subsets were stained: CD4+, CD8+, CD2+, CD25+, CD45RO+ and CD45RA+ T-cells. In addition cells expressing NK receptors were stained: CD94+ cells and CD161+ cells. T-cell subsets and cells expressing NK-receptors were analysed by immunohistochemical scoring. The proliferation marker Ki-67 and differentiation marker keratin-10 were revealed immunohistochemically by MIB-1 and RKSE60 respectively. Both markers were analysed using quantitative image analyses. The number of Ki-67 positive nuclei and the percentage of keratin-10 positive epidermal cells in flexural and plaque psoriasis were comparable. There is no difference between flexural and plaque psoriasis concerning other T-cell subsets. However a highly significant reduction is seen in flexural psoriasis with respect to CD161+ cells in the dermis. The similarity of chronic plaque psoriasis compared to flexural psoriasis with respect to T-cell subsets, epidermal proliferation and keratinization suggests that both conditions are pathogenetically identical. We propose the decreased quantity of lesional CD161+ cells in the dermis of flexural psoriatic lesions may result from chronic microbial challenge in flexural psoriasis.


Mirk protein kinase is activated by MKK3 and functions as a transcriptional activator of HNF1alpha.
Mirk/Dyrk1B is an arginine-directed serine/threonine protein kinase that is expressed at low levels in most normal tissues but at elevated levels in many tumor cell lines and in normal skeletal muscle. Colon carcinoma cell lines stably overexpressing Mirk proliferated in serum-free medium, but the mechanism of Mirk action is unknown. DCoHm (dimerization cofactor of hepatocyte nuclear factor 1alpha ( HNF1alpha) from muscle), a novel gene of the DCoH family with 78% amino acid identity to DCoH, was identified as a Mirk-binding protein by yeast two-hybrid analysis and cloned. Mirk co-immunoprecipitated with DCoHm and bound to DCoHm in glutathione S-transferase pull-down assays. DCoH stabilizes HNF1alpha as a dimer and enhances its transcriptional activity on the beta-fibrinogen promoter reporter, and DCoHm had similar activity. Mirk enhanced HNF1alpha transcriptional activity in a dose-dependent manner, whereas two kinase-inactive Mirk mutants and a Mirk N-terminal deletion mutant did not. Mirk, DCoHm, and HNF1alpha formed a complex. Mirk bound to a specific region within the CREB-binding protein-binding region of HNF1alpha and phosphorylated HNF1alpha at a site adjacent to the Mirk-binding region. Conversely, the HNF1alpha binding domain was located within the first five conserved kinase subdomains of Mirk. Mirk co-immunoprecipitated with the MAPK kinase MKK3, an upstream activator of p38. MKK3 enhanced Mirk kinase activity and the transcriptional activation of HNF1alpha by Mirk, suggesting that Mirk, like p38, is activated by certain environmental stress agents. The Mirk-binding protein DCoH has been shown to be selectively expressed in colon carcinomas but not in normal tissue. Mirk may function as an HNF1alpha transcriptional activator in response to an MKK3-mediated stress signal, and the selective expression of DCoH could restrict the Mirk response to carcinoma cells.


Au Octahedral Nanosponges: 3D Plasmonic Nanolenses for Near-Field Focusing.
Here, we report the synthesis of three-dimensional plasmonic nanolenses for strong near-field focusing. The nanolens exhibits a distinctive structural arrangement composed of nanoporous sponge-like networks within their interior. We denote these novel nanoparticles as "Au octahedral nanosponges" (Au Oh NSs). Employing a carefully planned multistep synthetic approach with Au octahedra serving as sacrificial templates, we successfully synthesized Au Oh NSs in solution. The porous domains resembling sponges contributed to enhanced scattering and absorption of incident light within metal ligaments. This optical energy was subsequently transferred to the nanospheres at the vertex, where near-field focusing was maximized. We named this observed enhancement a "lightning-sphere effect". Using single particle-by-particle surface-enhanced Raman scattering (SERS), we optimized the morphological dimensions of the spheres and porous domains to achieve the most effective near-field focusing. In the context of bulk SERS measurements targeting weakly adsorbing analytes (2-chloroethyl phenyl sulfide) in the gas phase, we achieved a low detection limit of 10 ppb. For nonadsorbing species (dimethyl methyl phosphonate), utilization of hybrid SERS substrates consisting of Au Oh NSs and metal-organic frameworks as gas-adsorbing intermediate layers was highly effective for successful SERS detection.


Epidemiologic evidence for time variation in HIV infectivity.
Data from three epidemiologic studies of heterosexual transmission of HIV among monogamous couples are used to assess evidence for time variation in HIV infectivity, possibly related to varying levels of infectiousness following infection in the primary infected partner. Analyses are based on statistical techniques that account for the inherent incompleteness of exposure information from such studies, and that allow direct assessment of the hypotheses that infectivity varies with time since infection and across partnerships. Data include findings from 302 couples from the California Partners' Study and 51 and 31 couples, respectively, from two U.S. Center for Disease Control and Prevention (CDC)-sponsored studies of infection in partners of transfusion recipients. Results indicate weak evidence for higher infectivity following infection of the primary partner, decreasing to relatively lower levels from 2 to 10 years after. Although these findings are consistent with biologic observations of time variation in viral levels, other explanations of the observed pattern (e.g., heterogeneity of infectivity) are equally plausible, pointing out some inherent limitations of data from such studies.


A rapid review of the effectiveness of screening practices at airports, land borders and ports to reduce the transmission of respiratory infectious diseases such as COVID-19.
Travel screening for infectious diseases is often implemented to delay or prevent the entry of infected persons to a country/area. To evaluate the effectiveness of different point-of-entry screening strategies in achieving a reduction in imported COVID-19 transmission. A rapid evidence review was conducted, systematically searching PubMed and Google Scholar and grey literature on 27 March 2020. We screened 1 194 records. Nine potential full-text articles were assessed for eligibility and included. Three articles investigated the effectiveness of entry-based thermal and body temperature scanning. Entry-based infrared thermal or body temperature scanning for COVID-19 was unlikely to be effective. Two systematic reviews found no additional benefit of travel restrictions/screening. In a COVID-19 modelling study, airport screening was not effective, with exit and entry thermal scanning identifying half and missing almost half of infected travellers. Two other modelling studies found that entry-based travel screening would achieve only modest delays in community transmission, while international travel quarantine could reduce case importations by 80%. There is insufficient evidence to support entry and exit screening at points of entry, as these strategies detect just over half of the infected cases, missing almost half at entry points. The benefits of airport screening therefore need to be context specific and weighed against the resources and cost of implementation, the contribution of imported cases to total cases, and the benefits of identifying 50% of cases in the South African context with the country's high HIV and tuberculosis prevalence and limited resources to deal with a pandemic of this nature.


Cloning and characterization of the mouse homologue of the human dendritic cell maturation marker CD208/DC-LAMP.
DC-LAMP, a member of the lysosomal-associated membrane protein (LAMP) family, is specifically expressed by human dendritic cells (DC) upon activation and therefore serves as marker of human DC maturation. DC-LAMP is detected first in activated human DC within MHC class II molecules-containing compartments just before the translocation of MHC class II-peptide complexes to the cell surface, suggesting a possible involvement in this process. The present study describes the cloning and characterization of mouse DC-LAMP, whose predicted protein sequence is over 50% identical to the human counterpart. The mouse DC-LAMP gene spans over 25 kb and shares syntenic chromosomal localization (16B2-B4 and 3q26) and conserved organization with the human DC-LAMP gene. Analysis of mouse DC-LAMP mRNA and protein revealed the expression in lung peripheral cells, but also its unexpected absence from mouse lymphoid organs and from mouse DC activated either in vitro or in vivo. In conclusion, mouse DC-LAMP is not a marker of mature mouse DC and this observation raises new questions regarding the role of human DC-LAMP in human DC.


Drosophila Rab2 controls endosome-lysosome fusion and LAMP delivery to late endosomes.
Rab2 is a conserved Rab GTPase with a well-established role in secretory pathway function and phagocytosis. Here we demonstrate that Drosophila Rab2 is recruited to late endosomal membranes, where it controls the fusion of LAMP-containing biosynthetic carriers and lysosomes to late endosomes. In contrast, the lysosomal GTPase Gie/Arl8 is only required for late endosome-lysosome fusion, but not for the delivery of LAMP to the endocytic pathway. We also find that Rab2 is required for the fusion of autophagosomes to the endolysosomal pathway, but not for the biogenesis of lysosome-related organelles. Surprisingly, Rab2 does not rely on HOPS-mediated vesicular fusion for recruitment to late endosomal membranes. Our work suggests that Drosophila Rab2 is a central regulator of the endolysosomal and macroautophagic/autophagic pathways by controlling the major heterotypic fusion processes at the late endosome.


Identifying and managing adverse environmental health effects: 2. Outdoor air pollution.
Air pollution contributes to preventable illness and death. Subgroups of patients who appear to be more sensitive to the effects of air pollution include young children, the elderly and people with existing chronic cardiac and respiratory disease such as chronic obstructive pulmonary disease and asthma. It is unclear whether air pollution contributes to the development of asthma, but it does trigger asthma episodes. Physicians are in a position to identify patients at particular risk of health effects from air pollution exposure and to suggest timely and appropriate actions that these patients can take to protect themselves. A simple tool that uses the CH2OPD2 mnemonic (Community, Home, Hobbies, Occupation, Personal habits, Diet and Drugs) can help physicians take patients' environmental exposure histories to assess those who may be at risk. As public health advocates, physicians contribute to the primary prevention of illness and death related to air pollution in the population. In this article we review the origins of air pollutants, the pathophysiology of health effects, the burden of illness and the clinical implications of smog exposure using the illustrative case of an adolescent patient with asthma.


Comparison of designs towards a subject-independent brain-computer interface based on motor imagery.
A major limitation of current Brain-Computer Interfaces (BCI) based on Motor Imagery (MI) is that they are subject-specific BCI, which require data recording and system training for each new user. This process is time consuming and inconvenient, especially for casual users or portable BCI with limited computational resources. In this paper, we explore the design of a Subject-Independent (SI) MI-based BCI, i.e., a BCI that can be used immediately by any new user without training the BCI with the user's data. This is achieved by training the BCI on data acquired from several other subjects. In order to assess the possibility to build such a BCI, we compared several designs based on different features and classifiers, on data from 9 subjects. Our results suggested that linear classifiers were the most appropriate for the design of MI-based SI-BCI. We also proposed a filter bank common spatial patterns feature extraction method based on a multi-resolution frequency decomposition which achieved the highest accuracy.


On-chip pressure sensor using single-layer concentric chambers.
A vision-based on-chip sensor for sensing local pressure inside a microfluidic device is proposed and evaluated in this paper. The local pressure is determined from the change of color intensity in the sensing chamber which is pre-filled with colored fluid. The working principle of the sensor is based on polydimethylsiloxane deformation. The pressure at the point of interest is guided into a deformation chamber, where the structural stiffness is softened by chamber geometry, and thus, the chamber deforms as a result of pressure changes. Such deformation is transmitted to the sensing chamber, a same-layer concentric inside the deformation chamber. The deformation in the sensing chamber causes the colored fluid flowing in or out the chamber and leads to different color intensity from the top view through a microscope. Experimental evaluations on static and dynamic responses by regulated input pressures were conducted. The correlation in static response is 0.97 while the dynamic responses are successfully observed up to 16 Hz. The greatest advantage is that the local pressure can be directly seen without any additional hardware or electricity. The whole sensor is on a single-layer microfluidic design, so that the fabrication is simple, consistent, and low-cost. The single-layer design also provides the convenience of easy integration for existing microfluidic systems.


Does language moderate the influence of information scanning and seeking on HPV knowledge and vaccine awareness and initiation among Hispanics?
To examine whether language moderates associations between three communication variables: media use, information scanning (attending to and remembering information) and seeking (actively looking for information), and three HPV outcomes: knowledge, vaccine awareness and vaccine initiation among Hispanics. Hispanic mothers of females aged 8-22 years (N=288) were surveyed. Univariate and multivariate logistic regressions investigated associations between communication variables and HPV outcomes. To examine moderation by language, we compared main effects and interaction models using the likelihood ratio test. For English- and Spanish-speakers, Internet use was associated with more HPV knowledge and vaccine awareness, but not initiation. Scanning and seeking were associated with more knowledge, vaccine awareness, and initiation. Language moderated effects of scanning and seeking only on vaccine awareness. Spanish speakers who scanned for information were more likely to be aware of the vaccine than those who did not (80% vs 26%); Spanish speakers who sought information were also more likely to be aware (95% vs 55%). For English speakers, vaccine awareness did not differ between those who scanned and sought and those who did not. Effects of information scanning and seeking on HPV vaccine awareness were much greater for Spanish than for English speakers. Providers, therefore, should not assume that Spanish-speaking mothers are already aware of the vaccine. Our findings call attention to heterogeneity within Hispanics which could be particularly important when examining health communication and cancer prevention behaviors.


Physiological recovery from firefighting activities in rehabilitation and beyond.
The primary objective of this study was to document the timeline of physiologic recovery from firefighting activities in order to inform emergency medical services (EMS) of vital sign values that might be expected during incident rehabilitation and in developing rehabilitation protocols to make decisions about when to return personnel to the fireground. Secondarily, we compared two different incident rehabilitation strategies to determine effectiveness in reducing physiologic strain following firefighting. A repeated-measures randomized crossover design was utilized in which firefighters conducted a controlled set of firefighting activities, after which they completed incident rehabilitation in one of two conditions: 1) similar to currently used rehabilitation protocols and 2) with active cooling and nutritional intervention. Following 15 minutes of rehabilitation, each firefighter was asked to perform a simulated rescue "dummy drag" and then recover for 120 minutes in a quiet area. Core temperature and heart rate were recorded throughout the study. Blood pressures and subendocardial viability ratios were obtained before firefighting, after firefighting, and at standardized times during rehabilitation and recovery. Heart rate and core temperature increased during firefighting, and core temperature continued to increase for 7 minutes after completion of firefighting activities. These values did not return to baseline until at least 50 minutes after firefighting activity. Systolic blood pressures were significantly reduced during rehabilitation (15.2%), and recovered 7.7% during the first 30 minutes of recovery, but remained significantly lower than before firefighting for at least 120 minutes. An index of subendocardial perfusion was also significantly depressed for up to 110 minutes after firefighters. Differences between rehabilitation protocols were minimal. The timeline for recovery from firefighting activities is significantly longer than the typical 10-20-minute rehabilitation period that often is provided on the fireground. Modifications from the current rehabilitation protocol do not appear to improve the recovery timeline when rehabilitation is conducted in a cool room. While firefighters often are concerned about elevated blood pressures, this study suggests that firefighters and EMS personnel should also be cognizant of the potential dangers of hypotension.


The ideal nucleoside/nucleotide backbone.
The selection of the nucleoside/nucleotide reverse transcriptase inhibitor (NRTI/NtRTI) backbone is an important step in constructing a regimen for the treatment of HIV infection. No single NRTI/NtRTI backbone has attributes that make it ideal for every patient, but each has its advantages. Data from clinical trials provide needed guidance on backbone selection and the choice of a 3rd agent to complete a highly active antiretroviral therapy (HAART) regimen. This article reviews available data published since 2000 that address NRTI/NtRTI backbone performance and safety when included in HAART regimens. Based on these trials, the characteristics of different options are compared with an ideal backbone and put into context with other considerations for successful regimen selection and improved outcomes in treatment-naive patients. Practical strategies are offered for implementing contemporary regimens in settings outside clinical trials.


Optimal Path Planning for Selective Waste Collection in Smart Cities.
Waste collection is one of the targets of smart cities. It is a daily task in urban areas and it entails the planning of waste truck routes, taking into account environmental, economic and social factors. In this work, an optimal path planning algorithm has been developed together with a practical software platform for smart and sustainable cities that enables computing the optimal waste collection routes, minimizing the impact, both environmental (CO2 emissions and acoustic damage) and socioeconomic (number of trucks to be used and fuel consumption). The algorithm is executed in Net2Plan, an open-source planning tool, typically used for modeling and planning communication networks. Net2Plan facilitates the introduction of the city layout input information to the algorithm, automatically importing it from geographical information system (GIS) databases using the so-called Net2Plan-GIS library, which can also include positions of smart bins. The algorithm, Net2Plan tool and its extension are open-source, available in a public repository. A practical case in the city of Cartagena (Spain) is presented, where the optimal path planning for plastic waste collection is addressed. This work contributes to the urban mobility plans of smart cities and could be extended to other smart cities scenarios with requests of optimal path planning.


A Fat to Forget: Trans Fat Consumption and Memory.
We sought to assess the relation of dietary trans fatty acid (dTFA) consumption to word-memory. We analyzed cross-sectional data from the 1999-2005 UCSD Statin Study. Participants were 1018 adult men and non-procreative women age ≥20 without diagnosed diabetes, CVD, or extreme LDL-cholesterol. Primary analyses focused on men, as only men (N = 694) were effectively represented in younger adult ages. "Recurrent words" assessed word memory. dTFA (grams/day) estimates were calculated from the Fred Hutchinson Food Frequency Questionnaire. Regression, stratified at age 45, assessed the relation between memory and dTFA in various adjustment models. Major findings were replicated in the full sample (including women). Potential mediators were examined. An age-by-dTFA interaction was significant. dTFA adversely predicted memory in younger adults (only), robust to adjustment model. Each gram/day dTFA was associated with an estimated 0.76 fewer words recalled (full model) (SE = 0.27, 95%CI = 0.22,1.3, P = 0.006). Adjustment for systolic blood pressure, waist circumference and BMI (but not lipid or glycemic variables) attenuated the relationship, consistent with mediation by factors involving, relating to, or concurrently influencing, these factors. Greater dTFA was significantly associated with worse word recall in younger adults. Prooxidant and energetic detriments of dTFA and triangulation with other evidence offer prospects for causality.


Identification of genes that contribute to the pathogenesis of invasive pneumococcal disease by in vivo transcriptomic analysis.
Streptococcus pneumoniae (the pneumococcus) continues to be responsible for a high level of global morbidity and mortality resulting from pneumonia, bacteremia, meningitis, and otitis media. Here we have used a novel technique involving niche-specific, genome-wide in vivo transcriptomic analyses to identify genes upregulated in distinct niches during pathogenesis after intranasal infection of mice with serotype 4 or 6A pneumococci. The analyses yielded 28 common, significantly upregulated genes in the lungs relative to those in the nasopharynx and 25 significantly upregulated genes in the blood relative to those in the lungs in both strains, some of which were previously unrecognized. The role of five upregulated genes from either the lungs or the blood in pneumococcal pathogenesis and virulence was then evaluated by targeted mutagenesis. One of the mutants (ΔmalX) was significantly attenuated for virulence in the lungs, two (ΔaliA and ΔilvH) were significantly attenuated for virulence in the blood relative to the wild type, and two others (ΔcbiO and ΔpiuA) were completely avirulent in a mouse intranasal challenge model. We also show that the products of aliA, malX, and piuA are promising candidates for incorporation into multicomponent protein-based pneumococcal vaccines currently under development. Importantly, we suggest that this new approach is a viable complement to existing strategies for the discovery of genes critical to the distinct stages of invasive pneumococcal disease and potentially has broad application for novel protein antigen discovery in other pathogens such as S. pyogenes, Haemophilus influenzae type b, and Neisseria meningitidis.


Cholera epidemic in Haiti, 2010: using a transmission model to explain spatial spread of disease and identify optimal control interventions.
Haiti is in the midst of a cholera epidemic. Surveillance data for formulating models of the epidemic are limited, but such models can aid understanding of epidemic processes and help define control strategies. To predict, by using a mathematical model, the sequence and timing of regional cholera epidemics in Haiti and explore the potential effects of disease-control strategies. Compartmental mathematical model allowing person-to-person and waterborne transmission of cholera. Within- and between-region epidemic spread was modeled, with the latter dependent on population sizes and distance between regional centroids (a "gravity" model). Haiti, 2010 to 2011. Haitian hospitalization data, 2009 census data, literature-derived parameter values, and model calibration. Dates of epidemic onset and hospitalizations. The plausible range for cholera's basic reproductive number (R(0), defined as the number of secondary cases per primary case in a susceptible population without intervention) was 2.06 to 2.78. The order and timing of regional cholera outbreaks predicted by the gravity model were closely correlated with empirical observations. Analysis of changes in disease dynamics over time suggests that public health interventions have substantially affected this epidemic. A limited vaccine supply provided late in the epidemic was projected to have a modest effect. Assumptions were simplified, which was necessary for modeling. Projections are based on the initial dynamics of the epidemic, which may change. Despite limited surveillance data from the cholera epidemic in Haiti, a model simulating between-region disease transmission according to population and distance closely reproduces reported disease patterns. This model is a tool that planners, policymakers, and medical personnel seeking to manage the epidemic could use immediately.


Moderate drop in water table increases peatland vulnerability to post-fire regime shift.
Northern and tropical peatlands represent a globally significant carbon reserve accumulated over thousands of years of waterlogged conditions. It is unclear whether moderate drying predicted for northern peatlands will stimulate burning and carbon losses as has occurred in their smaller tropical counterparts where the carbon legacy has been destabilized due to severe drainage and deep peat fires. Capitalizing on a unique long-term experiment, we quantify the post-wildfire recovery of a northern peatland subjected to decadal drainage. We show that the moderate drop in water table position predicted for most northern regions triggers a shift in vegetation composition previously observed within only severely disturbed tropical peatlands. The combined impact of moderate drainage followed by wildfire converted the low productivity, moss-dominated peatland to a non-carbon accumulating shrub-grass ecosystem. This new ecosystem is likely to experience a low intensity, high frequency wildfire regime, which will further deplete the legacy of stored peat carbon.


Improved culture flask for obligate anaerobes.
An improved flask system for the growth of extremely oxygen-sensitive bacteria in liquid culture is described. The improvement described utilizes an all-glass, neoprene-stoppered flask designed for growth of 50- to 1,000-ml cultures of bacteria with continuous gassing.


A decade-long analysis of gender, race, and ethnic representation of incoming cardiothoracic anesthesiology fellows.
This study calls attention to the necessity for increasing representation of minority groups among cardiothoracic fellowship programs. Demographics in medicine are changing, with more women and varying ethnicities contributing to the physician pool. Despite this change, diversity among cardiothoracic anesthesia fellows has not mirrored this trend. In November of 2022, medical student, anesthesia resident and cardiothoracic fellow demographics were analyzed from 2011 to 2021 to assess the changes in program diversity. Although male and female individuals have relatively similar representation in medical schools, female individuals only represent 34.8% of total US anesthesia residents. Cardiothoracic anesthesia fellowship programs continue to be male-sex dominant and white racial representation has consistently made up a majority of the total trainees. Increasing diversity amongst healthcare providers increases the likelihood of providing culturally competent care, thereby decreasing the healthcare gap in minority communities.


Reassessment of parameters for applying motor-unit triggered stimuli in peri-stimulus time histograms.
We reassessed the response properties of peri-stimulus time histograms (PSTHs) in cases when a test stimulus was triggered by a motor-unit discharge with a constant delay time. In this experiment, single motor unit action potentials were recorded from the right tibialis anterior (TA) muscle of five healthy persons. A test stimulus of the common peroneal nerve with low intensity to activate only Ia afferents of the TA was applied through a bipolar stimulating electrode placed distal to the neck of the fibula. We obtained several PSTHs with various delay times and stimulus intensities in the same recording session for maintaining the background property as the same among the test situations. As a result, we confirmed three characteristics of PSTHs from observed data: (1) given the same delay time (the same background firing properties), a weaker stimulus intensity evokes a lessened effect on PSTHs, naturally; (2) delay time alters the induction balance of direct and indirect effects on PSTHs even if the stimulus intensity is the same because the background firing properties are different; and (3) response probabilities do not correspond directly to stimulus intensities when background firing properties are different; it is possible for a relatively strong intensity stimulus to produce a weaker effect than a weak stimulus. We concluded that comparisons of effects taken at different phases in the control distribution (and also effects taken from different control distributions or different motor units) can be misleading. Therefore, such comparisons should only be made within data obtained from the same phase in the same control distribution of the same motor unit.


The feasibility of screening for intimate partner violence during postpartum home visits.
There is ongoing debate and limited evidence on the effectiveness of universal screening for intimate partner violence (IPV). The objectives of this descriptive qualitative study were to examine public health nurses' (PHNs') perceptions of screening for IPV; explore the feasibility, from the perspective of PHNs, of IPV screening during home visits; describe PHNs' screening practices; and describe PHN training in relation to IPV. Six PHNs discussed their experiences of addressing IPV both in the context of a randomized trial to evaluate screening methods and in the context of their general home visitation practices. The findings indicate that universal screening for IPV using a standard set of questions is difficult to implement during home visits to a general population of new mothers. For PHNs visiting high-risk families as part of the targeted Healthy Babies Healthy Children program, the standard practice is to assess for mothers' exposure to IPV during in-depth assessment of the family; the nature of in-depth assessment favours a case-finding rather than a screening approach to identifying women exposed to IPV.


Necrotizing enterocolitis after recurrent supraventricular tachycardia.
Necrotizing enterocolitis (NEC) is the most common gastrointestinal (GI) emergency that primarily affects premature infants. In the last decade, there have been significant advances to our understanding of neonatal NEC. The exact etiology of NEC remains unclear, but it is widely considered a multifactorial disease; prematurity, enteral feeding, intestinal hypoxia-ischemia, and bacterial colonization are considered major risk factors. We report a case of a 35 week preterm infant with recurrent episodes of supraventricular tachycardia who developed NEC in the second week of life. This case underscores the importance of being appropriately cautious in the feeding of these high-risk premature infants.


[SARS-CoV-2 Vaccination for Adult Patients with Inflammatory Bowel Disease: Expert Consensus Statements by KASID].
Coronavirus disease 2019 (COVID-19), caused by the novel coronavirus, is threatening global health worldwide with unprecedented contagiousness and severity. The best strategy to overcome COVID-19 is a vaccine. Various vaccines are currently being developed, and mass vaccination is in progress. Despite the very encouraging clinical trial results of these vaccines, there is insufficient information on the safety and efficacy of vaccines for inflammatory bowel disease (IBD) patients facing various issues. After reviewing current evidence and international guidelines, the Korean Association for the Study of Intestinal Diseases (KASID) developed an expert consensus statement on COVID-19 vaccination issues for Korean IBD patients. This expert consensus statement emphasizes that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination be strongly recommended for IBD patients, and it is safe for IBD patients receiving immunomodulatory therapy.


High school cross country running injuries: a longitudinal study.
To determine the incidence rate of injury among high school cross country runners over a 15-year period. Prospective-longitudinal. Twenty-three high schools in western Washington State under the surveillance of the University of Washington Athletic Health Care System between 1979-1994. One hundred and ninety-nine cross country teams. Injuries resulting from running in a cross country practice or meet. There were 1,622 injuries for an overall injury rate of 13.1/1,000 athletic exposures (AEs), i.e., participation of a runner in a practice or meet. Girls had a significantly higher overall injury rate (16.7/1,000 AEs) than boys (10.9/1,000 AEs) (p < 0.0001). Girls also had significantly higher injury rates than boys for both initial (p < 0.0001) and subsequent injuries (p < 0.0001), especially those at the same body location (p = 0.0001). This difference in risk estimates was consistent over a 15-year period. Nearly three-fourths of the injuries resulted in < or =4 days of disability. Overall, higher rates of initial injuries were reported during practices (9.2/1,000 AEs) than in meets (7.8/1,000 AEs) (p = 0.04). Shin injuries had the highest overall rates of new injury (1.9/1,000 AEs) and reinjury at the same body location (53.9/1,000 AEs). Girls had significantly higher initial injury rates than boys for shin (p < 0.0001), hip. and foot injuries (p < 0.01), and higher reinjury rates for knee. calf, and foot injuries, respectively (p < 0.05). The results of this study suggest that girl cross country runners are at higher risk of injury and reinjury than boy cross country runners.


Differential magnetic force microscope imaging.
This paper presents a method for differential magnetic force microscope imaging based on a two-pass scanning procedure to extract differential magnetic forces and eliminate or significantly reduce background forces with reversed tip magnetization. In the work, the difference of two scanned images with reversed tip magnetization was used to express the local magnetic forces. The magnetic sample was first scanned with a low lift distance between the MFM tip and the sample surface, and the magnetization direction of the probe was then changed after the first scan to perform the second scan. The differential magnetic force image was obtained through the subtraction of the two images from the two scans. The theoretical and experimental results have shown that the proposed method for differential magnetic force microscope imaging is able to reduce the effect of background or environment interference forces, and offers an improved image contrast and signal to noise ratio (SNR).


Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors.
We used recombinant SV40 (rSV40)-derived vectors to deliver transgenes to human and simian hematopoietic progenitor cells in culture, and in vivo after transduction ex vivo. rSV40 are highly efficient vectors that are made in very high titers. They infect almost all cells, whether resting or dividing. Two rSV40s were used: SV(HBS), carrying hepatitis B surface antigen as a marker; and SV(Aw) carrying IN#33, a single chain Fv antibody against HIV-1 integrase. CD34+ cells derived from human fetal bone marrow (HFBM) and rhesus macaque bone marrow were transduced once with SV(HBS) without selection. On average 60% of colonies derived from transduced CD34+ cells carried and expressed HBsAg, as assessed by PCR and immunochemistry. Transgene carriage persisted following differentiation of transduced rhesus CD34+ cells into T lymphocytes. In an effort to increase the percentage of gene-marked cells, three sequential treatments of CD34+ cells were done using SV(Aw), without selection. Two weeks later, >95% of colonies expressed IN#33. Unselected SV(Aw)-transduced CD34+ cells from HFBM were transplanted into sublethally irradiated SCID mice. Bone marrow harvested 3 months later showed that >50% of bone marrow cells expressed IN#33. This is comparable with the percentage of human cells in these animals' bone marrow as judged by immunostaining for human CD45. The stability and longevity of transduction in this setting suggests that rSV40 vectors integrate into the cellular genome. This possibility was supported by finding that PCR of genomic DNA using primer pairs with one cellular and one viral primer yielded PCR products only in transduced, but not control, cells. These PCR products hybridized with an SV40 DNA fragment. Thus, rSV40 vectors transduce normal human and primate bone marrow progenitor cells effectively without selection, and maintain transgene expression in vivo following reimplantation. Such high efficiency transduction may be useful in treating diseases of CD34+ cells and their derivatives.


Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton.
The capture of short-lived molecular states triggered by the early encounter of two or more interacting particles continues to be an experimental challenge of great interest to the field of cryo-electron microscopy (cryo-EM). A few methodological strategies have been developed that support these "time-resolved" studies, one of which, Spotiton-a novel robotic system-combines the dispensing of picoliter-sized sample droplets with precise temporal and spatial control. The time-resolved Spotiton workflow offers a uniquely efficient approach to interrogate early structural rearrangements from minimal sample volume. Fired from independently controlled piezoelectric dispensers, two samples land and rapidly mix on a nanowire EM grid as it plunges toward the cryogen. Potentially hundreds of grids can be prepared in rapid succession from only a few microliters of a sample. Here, a detailed step-by-step protocol of the operation of the Spotiton system is presented with a focus on troubleshooting specific problems that arise during grid preparation.


Evidence for a population of sleepy sodium channels in squid axon at low temperature.
We have studied the effects of temperature changes on Na currents in squid giant axons. Decreases in temperature in the 15-1 degrees C range decrease peak Na current with a Q10 of 2.2. Steady state currents, which are tetrodotoxin sensitive and have the same reversal potential as peak currents, are almost unaffected by temperature changes. After removal of inactivation by pronase treatment, steady state current amplitude has a Q10 of 2.3. Na currents generated at large positive voltages sometimes exhibit a biphasic activation pattern. The first phase activates rapidly and partially inactivates and is followed by a secondary slow current increase that lasts several milliseconds. Peak Na current amplitude can be increased by delivering large positive prepulses, an effect that is more pronounced at low temperatures. The slow activation phase is eliminated after a positive prepulse. The results are consistent with the hypothesis that there are two forms of the Na channel: (a) rapidly activating channels that completely inactivate, and (b) slowly activating "sleepy" channels that inactivate slowly if at all. Some fast channels are assumed to be converted to sleepy channels by cooling, possibly because of a phase transition in the membrane. The existence of sleepy channels complicates the determination of the Q10 of gating parameters and single-channel conductance.


Enzymatic functions of wild tomato methylketone synthases 1 and 2.
The trichomes of the wild tomato species Solanum habrochaites subsp. glabratum synthesize and store high levels of methylketones, primarily 2-tridecanone and 2-undecanone, that protect the plants against various herbivorous insects. Previously, we identified cDNAs encoding two proteins necessary for methylketone biosynthesis, designated methylketone synthase 1 (ShMKS1) and ShMKS2. Here, we report the isolation of genomic sequences encoding ShMKS1 and ShMKS2 as well as the homologous genes from the cultivated tomato, Solanum lycopersicum. We show that a full-length transcript of ShMKS2 encodes a protein that is localized in the plastids. By expressing ShMKS1 and ShMKS2 in Escherichia coli and analyzing the products formed, as well as by performing in vitro assays with both ShMKS1and ShMKS2, we conclude that ShMKS2 acts as a thioesterase hydrolyzing 3-ketoacyl-acyl carrier proteins (plastid-localized intermediates of fatty acid biosynthesis) to release 3-ketoacids and that ShMKS1 subsequently catalyzes the decarboxylation of these liberated 3-ketoacids, forming the methylketone products. Genes encoding proteins with high similarity to ShMKS2, a member of the "hot-dog fold" protein family that is known to include other thioesterases in nonplant organisms, are present in plant species outside the genus Solanum. We show that a related enzyme from Arabidopsis (Arabidopsis thaliana) also produces 3-ketoacids when recombinantly expressed in E. coli. Thus, the thioesterase activity of proteins in this family appears to be ancient. In contrast, the 3-ketoacid decarboxylase activity of ShMKS1, which belongs to the alpha/beta-hydrolase fold superfamily, appears to have emerged more recently, possibly within the genus Solanum.


Psychological factors underpinning vaccine willingness in Israel, Japan and Hungary.
The spread of SARS-CoV-2 led to rapid vaccine development. However, there remains considerable vaccine hesitancy in some countries. We investigate vaccine willingness in three nations with very different vaccine histories: Israel, Japan and Hungary. Employing an ecological-systems approach we analyse associations between health status, individual cognitions, norms, trust in government, COVID-19 myths and willingness to be vaccinated, with data from three nationally representative samples (Israel, Jan. 2021, N = 1011; Japan, Feb. 2021, N = 997; Hungary, April 2021, N = 1130). Vaccine willingness was higher in Israel (74%) than Japan (51%) or Hungary (31%). In all three countries vaccine willingness was greatest amongst who would regret not being vaccinated and respondents who trusted their government. Multi-group latent class analysis identified three groups of COVID myths, with particular concern about alteration of DNA (Israel), allergies (Hungary) and infection from the vaccine (Japan). Intervention campaigns should address such cultural myths while emphasising both individual and social benefits of vaccination.


Development and validation of an analytical method for the simultaneous determination of cocaine and its main metabolite, benzoylecgonine, in human hair by gas chromatography/mass spectrometry.
A new, simple and rapid procedure has been developed and validated for the determination of cocaine and its main metabolite, benzoylecgonine, in human hair samples. After extraction from within the hair matrix by a mixture of methanol/hydrochloric acid (2:1) at 65 degrees C for 3 h, and sample cleanup by mixed-mode solid-phase extraction (SPE), the extracts were analyzed by gas chromatography/mass spectrometry (GC/MS), after derivatization with N-methyl-N-(trimethylsilyl)trifluoroacetamide with 5% chlorotrimethylsilane. Using a sample size of only 20 mg of hair, limits of detection (LODs) and quantitation (LOQs) were, respectively, 20 and 50 pg/mg for cocaine, and 15 and 50 pg/mg for benzoylecgonine, achieving the cut-off values proposed by the Society of Hair Testing for the analysis of these compounds in hair. The method was found to be linear (weighing factor of 1/x) between the LOQ and 20 ng/mg for both compounds, with correlation coefficients ranging from 0.9974 to 0.9996 for cocaine; and from 0.9981 to 0.9994 for benzoylecgonine. Intra- and interday precision and accuracy were in conformity with the criteria normally accepted in bioanalytical method validation. The sample cleanup step presented a mean absolute recovery greater than 90% for both compounds. The developed method may be useful in forensic toxicology laboratories for the analysis of cocaine and benzoylecgonine in hair samples, taking into account its speed (only 3 h are required for the extraction of the analytes from within the matrix, whereas 5 h or even overnight extractions have been reported) and the low limits achieved (using a single quadrupole mass spectrometer, which is available in most laboratories).


Drivers of liking for yogurt drinks with prebiotics and probiotics.
Several studies have addressed the sensory properties of yogurt. However, as the market for yogurt continues to expand and new varieties of yogurt with novel ingredients emerge, additional sensory tests are needed to ensure the quality of the products. Three selected prebiotics, soluble corn fiber, polydextrose, and chicory inulin, were each added at an excellent source of fiber (5 g fiber/serving) or a good fiber source (2.5 g fiber/serving) levels into a yogurt drink base. Three additional yogurt drinks contained 5 g of each of the separate prebiotics along with a mixture of probiotics (Bifidobacterium lactis Bb-12 and Lactobacillus acidophilus LA-5). A control sample with no prebiotics or probiotics was also included in the experimental design. Yogurt drinks were evaluated by 110 consumers for overall acceptance, acceptance of aroma, appearance, taste, and texture, and purchase intent. Demographic information pertaining to consumer knowledge of prebiotics and probiotics was collected. Consumer data were correlated with previously obtained descriptive analysis data to identify drivers of liking. Data were analyzed by analysis of variance (ANOVA), Fisher's least significant difference (LSD), cluster analysis, internal preference mapping, and external preference mapping. Total variance explained by the internal and external preference maps were 32.2% and 64.6%, respectively, which showed higher levels of the prebiotics with probiotics drove consumer liking compared to lower levels without probiotics. In terms of ingredients added, chicory inulin and polydextrose were preferred over soluble corn fiber. Yogurt drinks with these prebiotics included and probiotics were characterized by a medium level of sweetness and high viscosity. Development of new prebiotic and probiotic containing drinkable yogurts should strive for a medium level of sweetness and high viscosity for maximum consumer acceptance.


Serum YKL-40 concentrations in patients with rheumatoid arthritis: relation to disease activity.
YKL-40, also called human cartilage glycoprotein-39, is secreted by chondrocytes, synovial cells, macrophages and neutrophils. Studies have shown that YKL-40 is an autoantigen in rheumatoid arthritis (RA). We evaluated whether serum YKL-40 was related to disease activity in patients with RA. Serum YKL-40 was determined by radioimmunoassay in 156 patients with RA during a 1 yr longitudinal study. Serum YKL-40 was increased in 54% of the patients with clinically active disease. Patients with clinically active disease initially who became inactive after 12 months had a significant decrease in serum YKL-40 (-30%, P < 0.002) and patients who changed from inactive to active disease had an increase in serum YKL-40. Patients who remained active had unchanged serum YKL-40 during the study. Serum YKL-40 decreased rapidly (-24% after 7 days, P < 0.01) during prednisolone therapy, and more slowly in patients treated with methotrexate only (-15% after 60 days, P < 0.01). Patients with early RA (disease duration < 3 yr, n = 50) and a persistently elevated serum YKL-40 were at risk of radiological disease progression as determined by Larsen score. Serum YKL-40 varies according to disease activity in RA, but provides in some respect information different from conventional markers. Our previous studies are consistent with a local release of YKL-40 in the arthritic joint followed by a secondary increase in serum YKL-40. YKL-40 may prove to be a new tool for the study of disease activity and pathophysiology of RA.


Postnatal alteration of collapsin response mediator protein 4 mRNA expression in the mouse brain.
Collapsin response mediator protein 4 (CRMP4) is a molecular marker for immature neurons but only limited information is available on the spatiotemporal gene expression changes of Crmp4 in the developing rodent. In the present study, the variation of CRMP4 mRNA expression in the mouse brain was investigated from postnatal day (PD) 0 (the day of birth) to adulthood by in situ hybridization. The hybridization signals were broadly detected on PD0 and regional changes in expression during development were noted. Expression patterns of CRMP4 mRNA were classified into the following three types: (i) signals that were strongest on PD0 or PD7, weak or undetectable on PD14, and absent in adulthood: this pattern was observed in most brain areas; (ii) signals that were first detected on PD0 or PD7 and persisted into adulthood: this pattern was seen in the dentate gyrus and subventricular zone of the olfactory bulb (OB); and (iii) signals that were strongest on PD0 and decreased gradually with age but were still detectable in adulthood: this pattern was identified for the first time in the mitral cell layer of the OB. Analysis using quantitative real-time RT-PCR confirmed higher expression of CRMP4 mRNA in the OB than in other adult brain regions. The persistence of CRMP4 mRNA in the adult OB, including the mitral cell layer, suggests the possibility of both neurogenetic and non-neurogenetic functional roles of CRMP4 in this region.


Creating frames of reference for chert exploitation during the Late Pleistocene in Southwesternmost Iberia.
Southwestern Iberia has played a key role in characterizing Late Pleistocene human ecodynamics. Among other aspects of human behavior, chert procurement and management studies in this region have received increasing attention in the past two decades, especially focusing on the sites showing repeated human occupation, such as the case of Vale Boi (Southern Portugal). However, these studies have been very limited in their geographical scope, and mostly focused on brief macroscopic descriptions of the raw materials. To further our knowledge of the relationship between regional availability of raw materials and its impact on human adaptations and mobility, a more detailed approach to characterizing geological sources is needed. This paper characterizes chert raw materials location, diversity, and availability in a geologically well-defined region of southern Portugal - the Algarve. Through macroscopic and petrographic approaches, we provide a detailed characterization of geological chert sources to build a frame of reference for chert exploitation in the region. Our results show that there are four main chert formations in Algarve, and that despite the within-source variability, sufficient differences at macroscopic and petrographic levels are present to allow clear source attribution. These results provide a baseline for raw material studies in archaeological assemblages across southwestern Iberia, that will be essential to further characterize the dynamics of human behavior in some of the most important eco-cultural niches.


Coordinate expression of Mn-containing superoxide dismutase and Cu,Zn-containing superoxide dismutase in human fibroblasts with trisomy 21.
The amount of Mn superoxide dismutase (MnSOD) and the activity of Cu,Zn-superoxide dismutase (CuZnSOD) have been studied in human fibroblasts of five subjects with trisomy 21 and five subjects with normal karyotype, using nuclear magnetic relaxation and polarographic methods. In the trisomic fibroblasts we have found a mean molar amount of MnSOD 25.4% lower than in the control, and an amount of CuZnSOD 54.7% higher. A positive significant correlation between the activities of both enzymes has been observed indicating that the two enzymes dismute the O2- cooperatively. However, the increase of MnSOD per unit of CuZnSOD appears significantly lower in the trisomic fibroblasts, an effect that is not due to a diminished inducibility of MnSOD. These findings suggest that the MnSOD and CuZnSOD genes interact to preserve the normal level of total SOD activity.


Young thrombocytes initiate the formation of arterial thrombi in zebrafish.
The zebrafish system is an excellent vertebrate genetic model to study hemostasis and thrombosis because saturation mutagenesis screens can identify novel genes that play a role in this vital physiologic pathway. To study hemostatic mutations, it is important to understand the physiology of zebrafish hemostasis and thrombosis. Previously, we identified zebrafish thrombocytes and have shown that they participate in arterial thrombus formation. Here, we recognized 2 populations of thrombocytes distinguishable by DiI-C18 (DiI) staining. DiI+ thrombocytes have a high density of adhesive receptors and are functionally more active than DiI- thrombocytes. We classified DiI+ thrombocytes as young and DiI- thrombocytes as mature thrombocytes. We found young and mature thrombocytes each formed independent clusters and that young thrombocytes clustered first. We have also shown that young thrombocytes initiate arterial thrombus formation. We propose that due to the increased adhesive receptor density on young thrombocytes, they adhere first to the subendothelial matrix, get activated rapidly, release agonists, and recruit more young thrombocytes, which further release more agonists. This increase in agonists activates the less active mature thrombocytes, drawing them to the growing thrombus. Since arterial thrombus formation is a fundamental hemostatic event, this mechanism may be conserved in mammals and may open new avenues for prevention of arterial thrombosis.


Purification and characterization of extracellular matrix-degrading metalloproteinase, matrin (pump-1), secreted from human rectal carcinoma cell line.
A metalloproteinase with Mr 29,000 was purified to homogeneity as a latent proenzyme from the conditioned medium of a human rectal carcinoma cell line CaR-1. This enzyme hydrolyzed casein more potently than gelatin embedded in polyacrylamide gels in zymography assay. Calcium ion was essential for the activity. It exerted the maximum activity at pH 7-9. Its activity was stimulated by organomercurials, such as p-amino-phenyl mercuric acetate and p-chloromercuric benzoic acid, and was inhibited by 1,10-phenanthroline but was hardly affected by diisopropyl fluorophosphate and pepstatin. When the purified proenzyme was activated by the organomercurials, it effectively hydrolyzed fibronectin, laminin, type IV basement membrane collagen, and several types of gelatins but not interstitial type I and III collagens. The treatment of the purified proenzyme with p-aminophenyl mercuric acetate or trypsin formed an active peptide with Mr 20,000. The structural analysis indicated that it was most likely identical to putative metalloproteinase-1, the complementary DNA of which had been cloned from human tumor mRNAs capable of hybridizing to a rat transin complementary DNA. Based on the fact that this enzyme is secreted extracellularly and degrades the matrix proteins, we propose the name "matrin" for this newly identified enzyme.


Impact of the fit and strong intervention on older adults with osteoarthritis.
This study assessed the impact of a low cost, multicomponent physical activity intervention for older adults with lower extremity osteoarthritis. A randomized controlled trial compared the effects of a facility-based multiple-component training program followed by home-based adherence (n = 80) to a wait list control group (n = 70). Assessments were conducted at baseline and at 2 and 6 months following randomization. The training program consisted of range of motion, resistance training, aerobic walking, and education-group problem solving regarding self-efficacy for exercise and exercise adherence. All training group participants developed individualized plans for posttraining adherence. Relative to the persons in the control group, individuals who participated in the exercise program experienced a statistically significant improvement in exercise efficacy, a 48.5% increase in exercise adherence, and a 13.3% increase in 6-min distance walk that were accompanied by significant decreases in lower extremity stiffness at 2 and 6 months. Program participants also experienced a significant decrease in lower extremity pain and a borderline significant improvement in efficacy to adhere to exercise over time at 6 months (p =.052). In contrast, persons in the control group deteriorated over time on the efficacy and adherence measures and showed no change on the other measures. No adverse health effects were encountered. These benefits indicate that this low-cost intervention may hold great promise as one of a growing number of public health intervention strategies for older adults in the United States with osteoarthritis.


Outcomes of damage control laparotomy with open abdomen management in the octogenarian population.
Controversy surrounds the role of abbreviated laparotomy and open abdomen (OA) in the octogenarian population in the acute care surgery model based on concern that the initial insult, combined with its sequelae, is beyond the physiologic reserve of these patients. As the population ages further, this dilemma will arise more frequently, requiring the analysis of futility or utility of OA in this demographic. The institutional review board approval was obtained to analyze retrospectively patients aged 80 years or older with OA from 1997 to 2009. Univariate, multivariate, and Kaplan-Meier analyses were used to evaluate the effects that demographics, comorbidities, and clinical factors had on in-hospital mortality and overall survival. Sixty-seven patients (32 men and 35 women) were identified. Acute general surgery (including vascular procedures) was the most common indication for laparotomy (94%) with trauma a distant second (6%). Early definitive closure was obtained in 52% of patients with a 34% planned ventral hernia rate. Overall complication rate was 62% and overall in-hospital mortality was 37%. Multivariate analysis revealed congestive heart failure (odds ratio, 11.4; 95% confidence interval, 1.01-128.03) and acute renal failure (odds ratio, 11.8; 95% confidence interval, 2.00-69.12) correlated with in-hospital mortality. Of those surviving to hospital dismissal, 2-year survival was 66% with a 17-month median follow-up (range, 1-125 months). There is utility in octogenarians undergoing aggressive surgical management that requires OA. These patients have high mortality rates, but long-term survival can be better than their peers with other chronic diseases if they survive the surgical insult. Patient selection should be based on preexisting comorbidities such as congestive heart failure and the development of acute renal failure. Despite the adequate long-term survival, most patients will leave the hospital with a hernia.


Genome-wide RNAi screen reveals a specific sensitivity of IRES-containing RNA viruses to host translation inhibition.
The widespread class of RNA viruses that utilize internal ribosome entry sites (IRESs) for translation include poliovirus and Hepatitis C virus. To identify host factors required for IRES-dependent translation and viral replication, we performed a genome-wide RNAi screen in Drosophila cells infected with Drosophila C virus (DCV). We identified 66 ribosomal proteins that, when depleted, specifically inhibit DCV growth, but not a non-IRES-containing RNA virus. Moreover, treatment of flies with a translation inhibitor is protective in vivo. Finally, this increased sensitivity to ribosome levels also holds true for poliovirus infection of human cells, demonstrating the generality of these findings.


A simplified method for the clinical-scale generation of central memory-like CD8+ T cells after transduction with lentiviral vectors encoding antitumor antigen T-cell receptors.
Adoptive transfer of antigen-specific CD8+ T cells can effectively treat patients with metastatic melanoma. Recent efforts have emphasized the in vitro generation of antitumor T cells by transduction of genes encoding antitumor T-cell receptors. At present, lentiviral vector-mediated transduction of CD8+ T cells relies on anti-CD3/CD28 bead stimulation; however, this method fails to efficiently expand CD8+ T cells. Herein we sought to establish a methodology for lentiviral vector transduction using optimal activating agents for efficient gene delivery and robust expansion of CD8+ T cells. To overcome the inability of anti-CD3/CD28 beads to efficiently expand CD8+ T cells, we evaluated alternative activating agents including feeder cells from allogeneic peripheral blood mononuclear cells and plate-bound anti-CD3 antibody. Analyses of gene transfer, cell phenotype, fold expansion, and biologic activities were used to determine the optimal methodology. Plate-bound anti-CD3 provided an ideal activation platform that afforded optimal lentiviral vector-mediated gene transfer efficiency (up to 90%), and coupled with peripheral blood mononuclear cells feeder cells yielded up to 600-fold expansion of CD8+ T cells within 12 days. The T-cell antigen receptor (TCR) engineered CD8+ T cells conferred specific antitumor activity and many displayed a central memory-like phenotype. The methodology described here could be readily applied for engineering CD8+ T cells with antitumor specificity for human adoptive immunotherapy.


Cloning and molecular characterisation of a Delta8-sphingolipid-desaturase from Nicotiana tabacum closely related to Delta6-acyl-desaturases.
Investigation on the absence of Delta(6)-desaturase activity in Nicotiana tabacum has led to the cloning of a new desaturase gene from this organism (NTDXDES) that exhibited unexpected biochemical activity. Cladistic analysis shows clustering of NTDXDES together with functional Delta(6)-acyl-desaturases of near Solanales plants, such as Borago and Echium. This group lies apart from that of previously characterised Delta(8)-sphingolipid-desaturases, which also includes two putative tobacco members identified in this study. Moreover, strong expression of NTDXDES is found in leaves, flowers, fruits and developing seeds of tobacco plants that is highly dependent on the development phase, with transcriptional activity being higher at stages of active tissue growth. This pattern is similar to that showed by Delta(6)-acyl-desaturases characterised in Boraginaceae species. However, functional assays using a yeast expression system revealed that the protein encoded by NTDXDES lacks Delta(6)-desaturase activity, but instead it is able to desaturate sphingolipid substrates by introducing a double bond on the Delta(8)-position. These data indicate that NTDXDES represent a novel desaturase gene placed in a different evolutionary lineage to that of previously characterised Delta(8)-desaturases.


Regulation of rat heart cytosol 5'-nucleotidase by adenylate energy charge.
A 5'-nucleotidase (EC 22.214.171.124) was highly purified from the soluble fraction of rat heart. The preparation appeared homogeneous by the criterion of polyacrylamide-gel electrophoresis. The enzyme was activated by ATP and ADP, and inhibited by Pi. When AMP was used as substrate, the velocity/substrate-concentration plot was sigmoidal. ATP or ADP changed the plot to hyperbolic and decreased S0.5. Pi increased both the sigmoidicity of the plot and S0.5. When IMP was used as substrate, the velocity/substrate plot was hyperbolic. ATP or ADP decreased Km and increased V. Pi changed the plot to sigmoidal and increased S0.5. Within the range of adenylate energy charge observed in surviving mammalian cells (0.7-0.9), the rate of AMP-hydrolysing activity catalysed by the 5'-nucleotidase increased sharply with decreasing energy charge. The highest activity was observed at an energy-charge value of about 0.6. The response was also observed in the presence of Pi. No change in IMP-hydrolysing activity was observed in the physiological range of adenylate energy charge, but in the presence of Pi the activity gradually increased with increasing energy charge. These results suggest the possibility that this enzyme participates in production of adenosine, a vasodilator, during hypoxia and in removal of IMP, which accumulates during the hypoxia, in the heart.


Immunohistochemical Localization of Translationally Controlled Tumor Protein in Axon Terminals of Mouse Hippocampal Neurons.
Translationally controlled tumor protein (TCTP) is a cytosolic protein with microtubule stabilization and calcium-binding activities. TCTP is expressed in most organs including the nervous system. However, detailed distribution and functional significance of TCTP in the brain remain unexplored. In this study, we investigated the global and subcellular distributions of TCTP in the mouse brain. Immunohistochemical analyses with anti-TCTP revealed that TCTP was widely distributed in almost all regions of the brain including the cerebral cortex, thalamus, hypothalamus, hippocampus, and amygdala, wherein it was localized in axon tracts and axon terminals. In the hippocampus, TCTP was prominently localized to axon terminals of the perforant path in the dentate gyrus, the mossy fibers in the cornu ammonis (CA)3 region, and the Schaffer collaterals in the CA1 field, but not in cell bodies of granule cells and pyramidal neurons, and in their dendritic processes. Widespread distribution of TCTP in axon tracts and axon terminals throughout the brain suggests that TCTP is likely involved in neurotransmitter release and/or maintaining synaptic structures in the brain, and that it might have a role in maintaining synaptic functions and synaptic configurations important for normal cognitive, stress and emotional functions.


Kringle IV Type 2, Not Low Lipoprotein(a), as a Cause of Diabetes: A Novel Genetic Approach Using SNPs Associated Selectively with Lipoprotein(a) Concentrations or with Kringle IV Type 2 Repeats.
Low plasma lipoprotein(a) concentrations are associated with type 2 diabetes. Whether this is due to low lipoprotein(a) concentrations per se or to a large number of kringle IV type 2 (KIV-2) repeats remains unclear. We therefore aimed to identify genetic variants associated selectively with lipoprotein(a) concentrations or with the number of KIV-2 repeats, to investigate which of these traits confer risk of diabetes. We genotyped 8411 individuals from the Copenhagen City Heart Study for 778 single-nucleotide polymorphisms (SNPs) in the proximity of the LPA gene, and examined the association of these SNPs with plasma concentrations of lipoprotein(a) and with KIV-2 number of repeats. SNPs that were selectively associated with lipoprotein(a) concentrations but not with KIV-2 number of repeats, or vice versa, were included in a Mendelian randomization study. We identified 3 SNPs (rs12209517, rs12194138, and rs641990) that were associated selectively with lipoprotein(a) concentrations and 3 SNPs (rs1084651, rs9458009, and rs9365166) that were associated selectively with KIV-2 number of repeats. For SNPs selectively associated with lipoprotein(a) concentrations, an allele score of 4-6 vs 0-2 had an odds ratio for type 2 diabetes of 1.03 (95% CI, 0.86-1.23). In contrast, for SNPs selectively associated with KIV-2 number of repeats, an allele score of 4-6 vs 0-2 had an odds ratio for type 2 diabetes of 1.42 (95% CI, 1.17-1.69). Using a novel genetic approach, our results indicate that it is a high number of KIV-2 repeats that are associated causally with increased risk of type 2 diabetes, and not low lipoprotein(a) concentrations per se. This is a reassuring finding for lipoprotein(a)-lowering therapies that do not increase the KIV-2 number of repeats.