Camptothecin and its derivatives induce expression of the c-jun protooncogene in human myeloid leukemia cells.
We have recently demonstrated that certain camptothecin derivatives are effective agents in the treatment of human tumor xenografts in nude mice. While camptothecin and its derivatives are recognized as inhibitors of topoisomerase I, little is known about the effects of these agents on specific gene expression, particularly genes involved in growth control. The c-jun early response gene codes for a leucine zipper transcription factor. The present studies demonstrate that 20(S)-camptothecin, 9-amino-20(S)-camptothecin, and 9-nitro-20(S)-camptothecin inhibit the growth of human U-937 myeloid leukemia cells and induce expression of the c-jun gene. c-jun transcripts were increased at 3 h and reached a maximum at 6 h of drug exposure. We also demonstrate that the induction of c-jun gene expression by these agents occurs at the transcriptional level. H7, a nonselective inhibitor of protein kinase C, completely blocked c-jun expression in 20(S)-camptothecin-treated cells, while another protein kinase inhibitor, HA1004, had no detectable effect. Similar findings were obtained for other leucine zipper encoding genes, including jun-B. These results suggest that 20(S)-camptothecin, 9-amino-20(S)-camptothecin, and 9-nitro-20(S)-camptothecin activate a cellular response involving the induction of early response genes. Finally, we demonstrate that induction of c-jun expression occurs in association with internucleosomal DNA fragmentation, a characteristic of programmed cell death.


Isolation and identification of two new diterpene glycosides from Nicotiana tabacum.
Two new diterpene glycosides containing 20-hydroxygeranyllinalool were isolated and identified from Nicotiana tabacum. These compounds consisted of five molecules of glucose and/or rhamnose. The locations of the aglycone and glycosides in the molecules were determined by 2D-NMR with the HMBC technique. The structures were (6E,10E,14Z)-20-hydroxygeranyllinalyl-3-O- [alpha-L-rhamnopyranosyl (1 --> 4)]-beta-D-glucopyranoside-20-O- [beta-D-glucopyranosyl (1 --> 2)]-[alpha-L-rhamnopyranosyl (1 --> 6)]-beta-D- glucopyranoside and (6E,10E,14Z)-20-hydroxygeranyllinalyl-3-O- [alpha-L-rhamnopyranosyl (1 --> 4)]-beta-D-glycopyranoside-20-O- [alpha-L-rhamnopyranosyl (1 --> 4)]-[alpha-L-rhamnopyranosyl (1 --> 6)]-beta-D- glucopyranoside.


Unveiling how an archetypal fluorescent protein operates: theoretical perspective on the ultrafast excited state dynamics of GFP variant S65T/H148D.
Green fluorescent protein variant S65T/H148D has been reported to host a photocycle involving the photoinduced proton transfer reaction between the chromophore and residue Asp148 under 50 fs and without a measurable kinetic isotope effect, and experimental evidence is suggestive of the existence of a highly delocalized proton between these residues. The blinding speed at which this biological system undergoes proton transfer has been ascribed to the extreme increase of acidity of the GFP chromophore in the electronic excited state where proton transfer takes place. This work strives to present a coherent, complete, and balanced description of the dynamics of this specific variant of GFP in which it will be shown that this increase of acidity is insufficient to explain the behavior observed. This study tracks the behavior of this photosystem to the delicate interplay between structure and dynamics shown in the presence of solvent. In this way, it has been found that the dynamics of this protein intertwines its structure with the intervening solvent to give rise to effectively degenerate situations in what concerns the reactants and products of the proton transfer reaction in ground and, most importantly, photoexcited state, in terms of potential energy profiles associated with the proton migration. Under these conditions, proton transfer can occur in accordance with the experimental data available. This set of characteristics is possibly common to a host of other proton transfer based fluorescent proteins, and helps promoting GFP S65T/H148D to a case of archetypal significance. Thus, our results can be useful to understand the way many fluorescent proteins work and, more generally, the molecular basis for proton transfer reactions in proteins.


Mapping human haematopoietic stem cells from haemogenic endothelium to birth.
The ontogeny of human haematopoietic stem cells (HSCs) is poorly defined owing to the inability to identify HSCs as they emerge and mature at different haematopoietic sites1. Here we created a single-cell transcriptome map of human haematopoietic tissues from the first trimester to birth and found that the HSC signature RUNX1+HOXA9+MLLT3+MECOM+HLF+SPINK2+ distinguishes HSCs from progenitors throughout gestation. In addition to the aorta-gonad-mesonephros region, nascent HSCs populated the placenta and yolk sac before colonizing the liver at 6 weeks. A comparison of HSCs at different maturation stages revealed the establishment of HSC transcription factor machinery after the emergence of HSCs, whereas their surface phenotype evolved throughout development. The HSC transition to the liver marked a molecular shift evidenced by suppression of surface antigens reflecting nascent HSC identity, and acquisition of the HSC maturity markers CD133 (encoded by PROM1) and HLA-DR. HSC origin was tracked to ALDH1A1+KCNK17+ haemogenic endothelial cells, which arose from an IL33+ALDH1A1+ arterial endothelial subset termed pre-haemogenic endothelial cells. Using spatial transcriptomics and immunofluorescence, we visualized this process in ventrally located intra-aortic haematopoietic clusters. The in vivo map of human HSC ontogeny validated the generation of aorta-gonad-mesonephros-like definitive haematopoietic stem and progenitor cells from human pluripotent stem cells, and serves as a guide to improve their maturation to functional HSCs.


[The elderly need better pain treatment].
Age distribution in developed countries is currently shifting towards older age. However, knowledge regarding diseases and treatment of the oldest patients remains incomplete, especially when it comes to acute and chronic pain. Pain assessment is complicated in these patients as they often have more than one diagnosis contributing to the clinical picture, in addition age-related changes in pain perception and cognitive impairment render evaluation of subjective symptoms difficult. Multidisciplinary pain management programmes that combine pharmacologic and non-pharmacologic treatment have proven efficacious in elderly patients. The choice of analgesics demands an understanding of age-related changes in drug-metabolism, co-morbidity and concurrent medication. Adjustment of drug doses and evaluation of benefits and risks should be tailored to each individual. There is an urgent need for better professional education programmes and further dedicated research specifically targeted to the needs of the elderly in our community.


Cataract and high-pass resolution perimetry.
The present study describes the influence of cataract on high-pass resolution perimetry results. Twenty-five otherwise healthy patients were examined before and after cataract surgery. Their preoperative visual acuities ranged from 0.1 to 0.8 and their mean resolution thresholds from 3.9 to 12.3 dB. Both elevated mean thresholds and local visual fields defects were observed. In patients with low-grade cataract, i.e. preoperative visual acuity 0.3 to 0.65, the mean improvement in perimetric results after operation was approximately 1 dB. In patients with higher degrees of lens opacity, visual acuity 0.1 to 0.25, the difference between pre- and postoperative perimetry results showed a wide range, 1.4-6.2 dB. In conclusion, cataract induced different types of visual field defects. The general threshold increase due to low-grade cataract (VA > or = 0.3) could be compensated for by subtracting 1 dB from the measured value.


Expression of Lubricin in the Human Amniotic Membrane.
Lubricin, a boundary lubricant, is the body's unique antiadhesive, antifibrotic, antifriction, and antiinflammatory glycoprotein. This amphiphile is produced by numerous tissues and acts to regulate a number of processes, such as homeostasis, shear stress, tissue development, innate immunity, inflammation, and wound healing. We hypothesize that lubricin is also synthesized and expressed by the amniotic membrane (AM), which also possesses antiadhesive, antifibrotic, and antiinflammatory properties. We also hypothesize that lubricin, at least in part, mediates these AM capabilities. Our goal was to test our hypothesis. We obtained multiple samples of fresh, cryopreserved (CP), and freeze-dried (FD) human AMs, as well as fresh placental tissue as positive controls, and processed them for light microscopy, immunofluorescence, and western blot analyses. We also evaluated the ability of recombinant human lubricin to associate with FD-AMs. Our results demonstrate that all fresh placental, fresh AM, and CP-AM samples contained lubricin. Lubricin was expressed in placental chorionic villi, AM epithelial and stromal cells, and CP-AM epithelia. No lubricin could be detected in FD-AMs but could be restored in FD-AMs after overnight incubation with recombinant human lubricin. This study supports our hypothesis that lubricin is expressed in human AMs. In addition, our data show that preservation methods influence the extent of this expression. Indeed, the disappearance of lubricin in FD-AMs may explain why dried AM reportedly loses its antiinflammatory and antiscarring abilities. It is possible that lubricin may mediate, at least in part, many of the biological properties of AMs.


Image guidance of breast cancer surgery using 3-D ultrasound images and augmented reality visualization.
This paper describes augmented reality visualization for the guidance of breast-conservative cancer surgery using ultrasonic images acquired in the operating room just before surgical resection. By combining an optical three-dimensional (3-D) position sensor, the position and orientation of each ultrasonic cross section are precisely measured to reconstruct geometrically accurate 3-D tumor models from the acquired ultrasonic images. Similarly, the 3-D position and orientation of a video camera are obtained to integrate video and ultrasonic images in a geometrically accurate manner. Superimposing the 3-D tumor models onto live video images of the patient's breast enables the surgeon to perceive the exact 3-D position of the tumor, including irregular cancer invasions which cannot be perceived by touch, as if it were visible through the breast skin. Using the resultant visualization, the surgeon can determine the region for surgical resection in a more objective and accurate manner, thereby minimizing the risk of a relapse and maximizing breast conservation. The system was shown to be effective in experiments using phantom and clinical data.


Astrocyte- and Microglia-Specific Mitochondrial DNA Deletions Levels in Sporadic Alzheimer's Disease.
Oxidative stress is implicated in the pathogenesis of neurodegenerative diseases, including sporadic Alzheimer's disease (AD). Mitochondrial DNA (mtDNA) deletions are markers of oxidative damage with an age-dependent accumulation. In a previous study, we analyzed mtDNA levels in diverse neuronal cell types in order to unravel the impact of oxidative stress in brains of AD patients. The aim of this study was to identify possible correlations between mtDNA deletion levels of selected astrocytes and microglia from three brain regions with different vulnerability to AD pathology and different stages of disease compared to controls. Our results reflect a higher vulnerability of hippocampal astrocytes and microglia to oxidative stress compared to other brain regions, such as cerebellum and brainstem.


Effects of grape seed proanthocyanidin extracts on peripheral nerves in streptozocin-induced diabetic rats.
Diabetic peripheral neuropathy (DPN) is one of the most common diabetic chronic complications. The aim of this study was to clarify whether grape seed proanthocyanidins extracts (GSPE) are therapeutic agents against DPN. In this study, we used streptozocin (STZ) to induce diabetic rats. GSPEs (250 mg/kg body weight/d) were administrated to diabetic rats for 24 wk. Motor nerve conductive velocity (MNCV) and mechanical hyperalgesia were determined in the rats. Serum glucose, glycated hemoglobin, advanced glycation end products (AGEs), and tissue malondialdehyde (MDA) and superoxide dismutase (SOD) were determined. Light and electron microscopy were used to observe the changes of nerval ultrastructure.GSPE significantly increased the MNCV, mechanical hyperalgesia and SOD of diabetic rats (p<0.05) and reduced the AGEs and MDA of diabetic rats (p<0.05). After being treated by GSPE, the severe segmental demyelination was decreased and Schwann cells were improved. In conclusion, GSPE plays an important role against DPN. With the decreasing of AGEs and MDA, it can ameliorate oxidation-associated nerval damage. This study may provide a new recognition of natural medicine for the treatment of DPN.


[Intra-arterial dynamic computed tomography in characterizing liver metastases of colorectal cancer].
Locoregional chemotherapy of liver metastases from colorectal tumours is a promising new approach. The results of CT in fifteen patients after intravenous and intra-arterial contrast injection are compared. The liver metastases are supplied mainly by the arterial route and this permits a means of assessing therapy. Contrast injections through a hepatic catheter may result in flow phenomena which limits the value of this investigation. It is unknown whether such flow phenomena may influence the result of intra-arterial chemotherapy. Although the margins of the metastases were more clearly defined, diagnostic accuracy was not increased, as compared with intravenous CT enhancement.


Enantio- and Diastereoselective Organocatalytic Conjugate Additions of Nitroalkanes to Enone Diesters.
Enantio- and diastereoselective conjugate addition reactions between nitroethane or nitropropane and enone diesters are reported. A bifunctional triaryliminophosphorane catalyzed the addition reaction with consistently excellent stereoselectivities and yields across a wide range of substrates. By the use of the gem-diester functional handle present in the adducts, local desymmetrization via diastereotopic group discrimination was demonstrated, and a polyfunctionalized lactam with three contiguous stereocenters was synthesized.


[Viremia in patients with tick-borne encephalitis and in persons with attached ixodes ticks].
An original method of virus isolation developed by the authors made it possible to collect 64 strains of tick-borne encephalitis (TBE) virus isolated from the blood of TBE patients and subjects complaining of sucking Ixodidae ticks. It is demonstrated that only a small part of the tick natural population cause acute TBE. The majority of the strains (54 isolates) of TBE virus have been isolated from the blood of clinically healthy subjects. Half of the strains showed no immunogenic properties in human body. Season infectivity corresponds to monthly virusophority of the ticks.


Ageing, depression, anxiety, social support and the diurnal rhythm and awakening response of salivary cortisol.
The present study compared the cortisol awakening response and diurnal rhythm in 24 young healthy students and 48 community-dwelling older adults. The associations with diurnal cortisol and depression, anxiety and social support were also examined in relation to age. Salivary cortisol was measured over the course of one day: immediately upon awakening, 30 min later, and then 3h, 6h, 9h and 12h post-awakening. Participants completed a questionnaire measuring symptoms of anxiety and depression and social support was assessed. Older adults exhibited a significantly reduced awakening response, overall cortisol levels, area under the curve (AUC) and diurnal slopes than younger adults, resulting in a flatter diurnal rhythm. Younger adults with higher depression scores had significantly higher overall cortisol and higher levels upon awakening and 30 min post-awakening. In the younger adults, anxiety and depression correlated positively with AUC and the cortisol awakening response (CAR). Older adults with lower social support had a reduced AUC where younger adults with lower social support displayed a larger AUC. These findings suggest that the diurnal rhythm and awakening response of salivary cortisol are significantly reduced in older adults and the associations between anxiety, depression and social support and diurnal cortisol vary with age.


Potential roles of ATP and local neurons in the monitoring of blood O2 content by rat aortic bodies.
Aortic bodies are arterial chemoreceptors presumed to monitor blood O2 content by unknown mechanisms, in contrast to their well-studied carotid body counterparts, which monitor PO2 and /pH. We recently showed that rat aortic body chemoreceptors (type I cells), located at the left vagus-recurrent laryngeal nerve bifurcation, responded to PO2 and PCO2 /pH in a manner similar to carotid body type I cells. These aortic bodies are uniquely associated with a group of local neurons, which are also sensitive to these stimuli. Here, we hypothesized that these local neurons may contribute to monitoring blood O2 content. During perforated patch recordings, ATP, known to be released from (carotid body) type I cells and red blood cells during hypoxia, induced inward currents and excited ≈ 45% of local neurons (EC50 ≈ 1 μm), mainly via heteromeric P2X2/3 purinoceptors. While ATP also induced a rise in intracellular [Ca(2+)] in a subpopulation of these neurons, almost all of them responded to nicotinic cholinergic agonists. During paired recordings, several juxtaposed neurons showed strong bidirectional electrical coupling, suggesting a local co-ordination of electrical activity. Perfusion with Evans Blue dye resulted in labelling of aortic body paraganglia, suggesting they have ready access to circulatory factors, e.g. ATP released from red blood cells during hypoxia. When combined with confocal immunofluorescence, the dye-labelled regions coincided with areas containing tyrosine hydroxylase-positive type I cell clusters and P2X2-positive nerve endings. We propose a working model whereby local neurons, red blood cells, ATP signalling and low blood flow contribute to the unique ability of the aortic body to monitor blood O2 content.


Ensuring financial access to hearing AIDS for infants and young children.
Many young children with permanent hearing loss do not receive hearing aids and related professional services, in part because of public and private financing limitations. In 2006 the Children's Audiology Financing Workgroup was convened by the National Center for Hearing Assessment and Management to evaluate and make recommendations about public and private financing of hearing aids and related professional services for 0- to 3-year-old children. The workgroup recommended 4 possible strategies for ensuring that all infants and young children with hearing loss have access to appropriate hearing aids and professional services: (1) clarify that the definition of assistive technology, which is a required service under Part C of the Individuals With Disabilities Education Act (IDEA), includes not only analog hearing aids but also digital hearing aids with appropriate features as needed by young children with hearing loss; (2) clarify for both state Medicaid and Children's Health Insurance Programs that digital hearing aids are almost always the medically necessary type of hearing aid required for infants and young children and should be covered under the Early and Periodic Screening, Diagnosis, and Treatment (EPSDT) program; (3) encourage the passage of private health insurance legislative mandates to require coverage of appropriate digital hearing aids and related professional services for infants and young children; and (4) establish hearing-aid loaner programs in every state. The costs of providing hearing aids to all 0- to 3-year old children in the United States are estimated here.


The human cathelicidin, LL-37, induces granzyme-mediated apoptosis in regulatory T cells.
LL-37 is a human cationic host defense peptide (antimicrobial peptide) belonging to the cathelicidin family of peptides. In this study, LL-37 was shown to kill stimulated and nonstimulated CD4(+)CD25(+)FoxP3(+) T cells (regulatory T cells; Tregs) through apoptosis, while having no cytotoxic effect on CD4(+)CD25(-) T cells at the same LL-37 concentrations. Of interest, Tregs were much more sensitive to LL-37 than many other cells, dying at 10-fold lower concentrations than other cell types tested. LL-37 exposure resulted in DNA fragmentation, chromatin condensation, and apoptotic body formation, all indicative of an apoptotic form of cell death. The importance of granzyme family members in the apoptosis of Tregs after LL-37 treatment was analyzed by using C57Bl/6 lymphocytes obtained from mice that were homozygous for null mutations in the granzyme B gene, and both the granzyme A and B genes. Granzyme A and granzyme B were both shown to play a role in LL-37-induced apoptosis of Tregs. Further analysis showed that apoptosis occurred primarily through caspase-dependent apoptosis at high LL-37 concentrations. However, grA-dependent/caspase-independent cell death was also observed. This suggests that LL-37 induces apoptosis in Tregs through multiple different mechanisms, initiated by the LL-37-induced leakage of granzymes from cytolytic granules. Our results imply that LL-37 administered at the site of a tumor could influence the adaptive antitumor immune response by killing Tregs and thus inhibiting their suppressor activity.


Signatures of a jet cocoon in early spectra of a supernova associated with a γ-ray burst.
Long γ-ray bursts are associated with energetic, broad-lined, stripped-envelope supernovae1,2 and as such mark the death of massive stars. The scarcity of such events nearby and the brightness of the γ-ray burst afterglow, which dominates the emission in the first few days after the burst, have so far prevented the study of the very early evolution of supernovae associated with γ-ray bursts3. In hydrogen-stripped supernovae that are not associated with γ-ray bursts, an excess of high-velocity (roughly 30,000 kilometres per second) material has been interpreted as a signature of a choked jet, which did not emerge from the progenitor star and instead deposited all of its energy in a thermal cocoon4. Here we report multi-epoch spectroscopic observations of the supernova SN 2017iuk, which is associated with the γ-ray burst GRB 171205A. Our spectra display features at extremely high expansion velocities (around 115,000 kilometres per second) within the first day after the burst5,6. Using spectral synthesis models developed for SN 2017iuk, we show that these features are characterized by chemical abundances that differ from those observed in the ejecta of SN 2017iuk at later times. We further show that the high-velocity features originate from the mildly relativistic hot cocoon that is generated by an ultra-relativistic jet within the γ-ray burst expanding and decelerating into the medium that surrounds the progenitor star7,8. This cocoon rapidly becomes transparent9 and is outshone by the supernova emission, which starts to dominate the emission three days after the burst.


The crosstalk between gut inflammation and gastrointestinal disorders during acute pancreatitis.
The intestinal inflammation caused by intestinal ischemia reperfusion during acute pancreatitis (AP) often leads to multiple organ dysfunction and aggravation of acute pancreatitis. This review concerns up-date progress of the pathophysiology and molecular mechanism of the excessive production of gut-derived cytokines. The regulation effects of immuno-neuro-endocrine network for pancreatic necrosis are the basis for pharmacological therapeutic in AP. The translation from basic research to clinical trials for the prevention or treatment of severe acute pancreatitis (SAP) is of great value. Early enteral nutrition is necessary for the restitution, proliferation, and differentiation of the intestinal epithelial cells adjacent to the wounded area. Clearance of the excess intestinal bacteria and supplement of probiotics may be helpful to prevent bacterial translocation and infection of pancreas.


Quad-barrel multifunctional electrochemical and ion conductance probe for voltammetric analysis and imaging.
The fabrication and use of a multifunctional electrochemical probe incorporating two independent carbon working electrodes and two electrolyte-filled barrels, equipped with quasi-reference counter electrodes (QRCEs), in the end of a tapered micrometer-scale pipet is described. This "quad-probe" (4-channel probe) was fabricated by depositing carbon pyrolytically into two diagonally opposite barrels of a laser-pulled quartz quadruple-barrelled pipet. After filling the open channels with electrolyte solution, a meniscus forms at the end of the probe and covers the two working electrodes. The two carbon electrodes can be used to drive local electrochemical reactions within the meniscus while a bias between the QRCEs in the electrolyte channels provides an ion conductance signal that is used to control and position the meniscus on a surface of interest. When brought into contact with a surface, localized high resolution amperometric imaging can be achieved with the two carbon working electrodes with a spatial resolution defined by the meniscus contact area. The substrate can be an insulating material or (semi)conductor, but herein, we focus mainly on conducting substrates that can be connected as a third working electrode. Studies using both aqueous and ionic liquid electrolytes in the probe, together with gold and individual single walled carbon nanotube samples, demonstrate the utility of the technique. Substrate generation-dual tip collection measurements are shown to be characterized by high collection efficiencies (approaching 100%). This hybrid configuration of scanning electrochemical microscopy (SECM) and scanning electrochemical cell microscopy (SECCM) should be powerful for future applications in electrode mapping, as well as in studies of insulating materials as demonstrated by transient spot redox-titration measurements at an electrostatically charged Teflon surface and at a pristine calcite surface, where a functionalized probe is used to follow the immediate pH change due to dissolution.


A bacterial artificial chromosome library for soybean PI 437654 and identification of clones associated with cyst nematode resistance.
We have constructed a soybean bacterial artificial chromosome (BAC) library using the plant introduction (PI) 437654. The library contains 73 728 clones stored in 192 384-well microtiter plates. A random sampling of 230 BACs indicated an average insert size of 136 kb with a range of 20 to 325 kb, and less than 4% of the clones do not contain inserts. Ninety percent of BAC clones in the library have an average insert size greater than 100 kb. Based on a genome size of 1115 Mb, library coverage is 9 haploid genome equivalents. Screening the BAC library colony filters with cpDNA sequences showed that contamination of the genomic library with chloroplast clones was low (1.85%). Library screening with three genomic RFLP probes linked to soybean cyst nematode (SCN) resistance genes resulted in an average of 18 hits per probe (range 7 to 30). Two separate pools of forward and reverse suppression subtractive cDNAs obtained from SCN-infected and uninfected roots of PI437654 were hybridized to the BAC library filters. The 488 BACs identified from positive signals were fingerprinted and analyzed using FPC software (version 4.0) resulting in 85 different contigs. Contigs were grouped and analyzed in three categories: (1) contigs of BAC clones which hybridized to forward subtracted cDNAs, (2) contigs of BAC clones which hybridized to reverse subtracted cDNAs, and (3) contigs of BAC clones which hybridized to both forward and reverse subtracted cDNAs. This protocol provides an estimate of the number of genomic regions involved in early resistance response to a pathogenic attack.


Adaptation to intermittent positive pressure ventilation applied through the nose during day and night.
A 49 yr old poliomyelitic patient had been under cuirass-type nocturnal negative pressure ventilation for more than 20 yrs. He had a severe restrictive ventilatory impairment, and normal awake blood gases at rest and during light exercise. He was offered a trial of intermittent positive pressure ventilation applied through the nose (nIPPV). Two daytime studies and one night study were carried out under nIPPV, and one night study was performed under negative pressure ventilation. Tidal volume, respiratory frequency (Respitrace), blood gases and electromyogram (EMG) of the diaphragm (DEMG, oesophageal electrode) and/or sternocleidomastoid (ScEMG, surface electrodes) were measured. During daytime studies under nIPPV, the DEMG (and/or the ScEMG) did not decrease by more than 25% (p less than 0.005). However, when the patient was encouraged to relax, the DEMG decreased by 62% (p less than 0.001). Tidal volume and ventilation significantly increased during daytime nIPPV (p less than 0.025), whereas blood gases were kept at physiological levels. At night, the ScEMG was present and prominent until sleep onset. Thereafter it disappeared and remained silent, including periods of wakefulness during sleep time, until final awakening in the morning. This was true for both negative pressure ventilation and nIPPV. Snoring was present throughout sleep under negative pressure ventilation but not under nIPPV. We conclude that the behavioural response of the subject may determine the electrical activity of respiratory muscles during assisted ventilation.


Acute epithelial injury in the rat small intestine in vivo is associated with expanded expression of transforming growth factor alpha and beta.
Previous studies have shown the importance of transforming growth factors alpha and beta (TGF alpha and TGF beta) in modulating epithelial cell restitution after injury in vitro. To investigate the role of the growth factors TGF alpha and TGF beta after acute epithelial injury in vivo. An in vivo model of phytohaemagglutinin (PHA) induced acute epithelial injury in rat small intestine was used. Epithelial cell turnover was assessed by autoradiography and liquid scintillation counting of thymidine uptake. Expression of TGF alpha and TGF beta was assessed by immunohistochemistry. An expansion of the proliferative compartment and increased turnover of intestinal epithelial cells was seen in rats with PHA induced intestinal epithelial injury. Expression of TGF alpha and TGF beta peptides was shown in both the epithelial cell and lamina propria compartment. Different patterns of TGF alpha and TGF beta expression were seen, however, within the epithelium of rats with acute intestinal injury compared with untreated controls, while the expression of these peptides within the lamina propria was not changed. These findings suggest that acute intestinal epithelial cell injury in vivo is associated with compensatory changes in expression of TGF alpha and TGF beta in the epithelial cell compartment, while the lamina propria does not seem to be significantly affected.


Attitudes toward group-based inequality: social dominance or social identity?
In five studies we explored how the context in which people think about the social structure and the implications of the social structure for one's in-group affect attitudes toward inequality. In Studies 1 and 2 we found that social dominance orientation (SDO) scores reflect attitudes toward specific types of inequality that are salient in context. Consistent with social identity theory, in Studies 3 to 5 we found that SDO scores reflected the interests of specific group identities. Indeed, when we compared existing privileged and disadvantaged groups, and when we manipulated in-group status, we found that participants held more positive attitudes toward inequality when the in-group was privileged, compared to when the in-group was disadvantaged. Across all of our studies, results were consistent with the contention that attitudes toward inequality are group-specific and depend on the social-structural position of salient in-groups. We discuss the implications of our findings for social dominance theory.


The Long-Term Effects of Early Life Stress on the Modulation of miR-19 Levels.
MicroRNAs (miRNAs), one of the major small non-coding RNA classes, have been proposed as regulatory molecules in neurodevelopment and stress response. Although alterations in miRNAs profiles have been implicated in several psychiatric and neurodevelopmental disorders, the contribution of individual miRNAs in brain development and function is still unknown. Recent studies have identified miR-19 as a key regulator of brain trajectories, since it drives the differentiation of neural stem cells into mature neurons. However, no findings are available on how vulnerability factors for these disorders, such as early life stress (ELS), can modulate the expression of miR-19 and its target genes. To reach our aim, we investigated miR-19 modulation in human hippocampal progenitor stem cells (HPCs) treated with cortisol during 3 days of proliferation and harvested immediately after the end of the treatment or after 20 days of differentiation into mature neurons. We also analyzed the long-term expression changes of miR-19 and of its validated target genes, involved in neurodevelopment and inflammation, in the hippocampus of adult rats exposed or not to prenatal stress (PNS). Interestingly, we observed a significant downregulation of miR-19 levels both in proliferating (FC = -1.59, p-value = 0.022 for miR-19a; FC = -1.79, p-value = 0.016 for miR-19b) as well as differentiated HPCs (FC = -1.28, p-value = 0.065 for miR-19a; FC = -1.75, p-value = 0.047 for miR-19b) treated with cortisol. Similarly, we found a long-term decrease of miR-19 levels in the hippocampus of adult PNS rats (FC = -1.35, p-value = 0.025 for miR-19a; FC = -1.43, p-value = 0.032 for miR-19b). Among all the validated target genes, we observed a significant increase of NRCAM (FC = 1.20, p-value = 0.027), IL4R (FC = 1.26, p-value = 0.046), and RAPGEF2 (FC = 1.23, p-value = 0.020).We suggest that ELS can cause a long-term downregulation of miR-19 levels, which may be responsible of alterations in neurodevelopmental pathways and in immune/inflammatory processes, leading to an enhanced risk for mental disorders later in life. Intervention strategies targeting miR-19 may prevent alterations in these pathways, reducing the ELS-related effects.


Operando X-ray absorption and EPR evidence for a single electron redox process in copper catalysis.
An unprecedented single electron redox process in copper catalysis is confirmed using operando X-ray absorption and EPR spectroscopies. The oxidation state of the copper species in the interaction between Cu(ii) and a sulfinic acid at room temperature, and the accurate characterization of the formed Cu(i) are clearly shown using operando X-ray absorption and EPR evidence. Further investigation of anion effects on Cu(ii) discloses that bromine ions can dramatically increase the rate of the redox process. Moreover, it is proven that the sulfinic acids are converted into sulfonyl radicals, which can be trapped by 2-arylacrylic acids and various valuable β-keto sulfones are synthesized with good to excellent yields under mild conditions.


[Effect of the B-group vitamin complex on the blood content of saturated and unsaturated fatty acids in patients with ischemic heart disease and hypertension].
Gas-liquid chromatography was used to study the blood content of saturated and unsaturated fatty acids, under the influence of the functionally-associated vitamin-B complex, in 45 patients with coronary heart disease and essential hypertension. The vitamins were given daily in the following doses: thiamine diphosphate 50 mg, riboflavine 40 mg, calcium pantothenate 200 mg, nicotinic acid 200 mg and lipoic acid 50 mg. Favourable shifts leading to positive clinical effects were recorded in the fatty acid metabolism after 10-day taking the vitamin-B complex: the content of unsaturated (linoleic and arachidonic) fatty acids increased while that of saturated (stearic and palmitic) fatty acids decreased.


The impact of group psychological abuse on distress: the mediating role of social functioning and resilience.
Previous studies indicate that social functioning and resilience can mitigate the adverse psychological effects of interpersonal violence. Unfortunately, the role of these variables has not been studied in survivors of groups, organizations, and communities in which psychological abusive strategies are inflicted to recruit and dominate their members. To examine the mediating role of social functioning and resilience in the relationship between psychological abuse experienced in the past while in a group and current psychosocial distress and psychopathological symptoms. In this cross-sectional study, an online questionnaire was administered to 794 English-speaking former members of different kinds of groups, such as religious, pseudo therapeutic, pyramid scheme groups, and others. Among them, 499 were victims of group psychological abuse and 295 were non-victims. Victims of group psychological abuse reported lower levels of social functioning and resilience than non-victims, and higher levels of psychosocial difficulties and psychopathological symptoms. Serial mediation analyses revealed that social functioning and resilience mediated part of the impact of group psychological abuse on psychosocial difficulties and psychopathological symptoms. Sex and age joining the group were included as covariates. Participants who had experienced higher levels of group psychological abuse tend to have poorer social functioning, which is related to lower resilience. In turn, lower levels of social functioning and resilience are related with higher distress. This research sheds light on the underlying mechanisms involved in the relationship between group psychological abuse and distress suffered following this kind of traumatic experiences. Findings highlight the protective role of social adjustment, which can help promote and enhance resilience and mitigate psychosocial difficulties and psychopathological symptoms in survivors of group psychological abuse.


Mindfulness practice predicts interleukin-6 responses to a mindfulness-based alcohol relapse prevention intervention.
Chronic alcohol misuse can result in chronically elevated interleukin (IL)-6, a pro-inflammatory cytokine, in the bloodstream. Given that Mindfulness-Based Relapse Prevention (MBRP) has been shown to reduce alcohol misuse, MBRP might also be effective in reducing IL-6 concentrations. Past research has found, however, that IL-6 does not respond consistently to mindfulness-based interventions. Building on prior studies, we examined whether between-person variability in engagement with mindfulness training (i.e., formal mindfulness practice time) is associated with between-person variability in changes in serum IL-6, using data from a randomized controlled trial evaluating MBRP for Alcohol Dependence (MBRP-A). Participants were 72 alcohol dependent adults (mean age = 43.4 years, 63.9% male, 93.1% White) who received a minimum dose (i.e., at least four sessions) of MBRP-A either at the start of the trial (n = 46) or after a 26-week delay (n = 26). IL-6 concentrations did not significantly change from pre- to post-intervention for the full sample. Nevertheless, greater mindfulness practice time was significantly associated with reduced IL-6 levels (r = -0.27). The association between practice time and IL-6 changes remained significant when controlling for intervention timing (i.e., immediate or after the 26-week delay), demographic characteristics, and changes in mindful awareness, obsessive-compulsive drinking, and depressive symptoms. The association between practice time and IL-6 changes was not significant when omitting the minimum treatment dose requirement. Overall, results suggest that the level of engagement in mindfulness training may predict changes in the inflammatory pathophysiology in adults with alcohol dependence.


Cooperative binding of monodisperse anionic amphiphiles to the i-face: phospholipase A2-paradigm for interfacial binding.
Equilibrium parameters for the binding of monodisperse alkyl sulfate along the i-face (the interface binding surface) of pig pancreatic IB phospholipase A(2) (PLA2) to form the premicellar complexes (E(i)(#)) are characterized to discern the short-range specific interactions. Typically, E(i)(#) complexes are reversible on dilution. The triphasic binding isotherm, monitored as the fluorescence emission from the single tryptophan of PLA2, is interpreted as a cooperative equilibrium for the sequential formation of three premicellar complexes (E(i)(#), i = 1, 2, 3). In the presence of calcium, the dissociation constant K(1) for the E(1)(#) complex of PLA2 with decyl sulfate (CMC = 4500 microM) is 70 microM with a Hill coefficient n(1) = 2.1 +/- 0.2; K(2) for E(2)(#) is 750 microM with n(2) = 8 +/- 1, and K(3) for E(3)(#) is 4000 microM with an n(3) value of about 12. Controls show that (a) self-aggregation of decyl sulfate alone is not significant below the CMC; (b) occupancy of the active site is not necessary for the formation of E(i)(#); (c) K(i) and n(i) do not change significantly due to the absence of calcium, possibly because alkyl sulfate does not bind to the active site of PLA2; (d) the E(i)(#) complexes show a significant propensity for aggregation; and (e) PLA2 is not denatured in E(i)(#). The results are interpreted to elaborate the model for atomic level interactions along the i-face: The chain length dependence of the fit parameters suggests that short-range specific anion binding of the headgroup is accompanied by desolvation of the i-face of E(i)(#). We suggest that allosteric activation of PLA2 results from such specific interactions of the amphiplies and the desolvation of the i-face. The significance of these primary interfacial binding events and the coexistence of the E and E(i)(#) aggregates is discussed.


State Intimate Partner Violence-Related Firearm Laws and Intimate Partner Homicide Rates in the United States, 1991 to 2015.
To prevent intimate partner homicide (IPH), some states have adopted laws restricting firearm possession by intimate partner violence (IPV) offenders. "Possession" laws prohibit the possession of firearms by these offenders. "Relinquishment" laws prohibit firearm possession and also explicitly require offenders to surrender their firearms. Few studies have assessed the effect of these policies. To study the association between state IPV-related firearm laws and IPH rates over a 25-year period (1991 to 2015). Panel study. United States, 1991 to 2015. Homicides committed by intimate partners, as identified in the Federal Bureau of Investigation's Uniform Crime Reports, Supplementary Homicide Reports. IPV-related firearm laws (predictor) and annual, state-specific, total, and firearm-related IPH rates (outcome). State laws that prohibit persons subject to IPV-related restraining orders from possessing firearms and also require them to relinquish firearms in their possession were associated with 9.7% lower total IPH rates (95% CI, 3.4% to 15.5% reduction) and 14.0% lower firearm-related IPH rates (CI, 5.1% to 22.0% reduction) than in states without these laws. Laws that did not explicitly require relinquishment of firearms were associated with a non-statistically significant 6.6% reduction in IPH rates. The model did not control for variation in implementation of the laws. Causal interpretation is limited by the observational and ecological nature of the analysis. Our findings suggest that state laws restricting firearm possession by persons deemed to be at risk for perpetrating intimate partner abuse may save lives. Laws requiring at-risk persons to surrender firearms already in their possession were associated with lower IPH rates. Robert Wood Johnson Foundation.


Bayesian sample size for diagnostic test studies in the absence of a gold standard: Comparing identifiable with non-identifiable models.
Diagnostic tests rarely provide perfect results. The misclassification induced by imperfect sensitivities and specificities of diagnostic tests must be accounted for when planning prevalence studies or investigations into properties of new tests. The previous work has shown that applying a single imperfect test to estimate prevalence can often result in very large sample size requirements, and that sometimes even an infinite sample size is insufficient for precise estimation because the problem is non-identifiable. Adding a second test can sometimes reduce the sample size substantially, but infinite sample sizes can still occur as the problem remains non-identifiable. We investigate the further improvement possible when three diagnostic tests are to be applied. We first develop methods required for studies when three conditionally independent tests are available, using different Bayesian criteria. We then apply these criteria to prototypic scenarios, showing that large sample size reductions can occur compared to when only one or two tests are used. As the problem is now identifiable, infinite sample sizes cannot occur except in pathological situations. Finally, we relax the conditional independence assumption, demonstrating in this once again non-identifiable situation that sample sizes may substantially grow and possibly be infinite. We apply our methods to the planning of two infectious disease studies, the first designed to estimate the prevalence of Strongyloides infection, and the second relating to estimating the sensitivity of a new test for tuberculosis transmission. The much smaller sample sizes that are typically required when three as compared to one or two tests are used should encourage researchers to plan their studies using more than two diagnostic tests whenever possible. User-friendly software is available for both design and analysis stages greatly facilitating the use of these methods.


A long-lived iridium(iii) chemosensor for the real-time detection of GHB.
Gamma-hydroxybutyric acid (GHB) is a commonly used "date rape" drug in drug-facilitated sexual assaults (DFSA) due to its low price and ready availability, and it is more quickly excreted from the human body than other drugs. Therefore, the development of a real-time detection method for GHB could help facilitate law enforcement agencies in tackling DFSA. In this work, a long-lived iridium(iii) chemosensor 1 has been synthesized for the switch-off detection of GHB. Upon addition of GHB to an aqueous solution of the iridium(iii) complex 1, a significant luminescence quenching occurred that could be observed by the naked eye under UV illumination. The luminescence signal of the iridium(iii) complex 1 could also be distinguished from strongly fluorescent media using time-resolved emission spectroscopy.


Mechanistic studies of anticancer aptamer AS1411 reveal a novel role for nucleolin in regulating Rac1 activation.
AS1411 is a G-rich quadruplex-forming oligodeoxynucleotide that binds specifically to nucleolin, a protein found on the surface and in the cytoplasm of most malignant cells but absent from the surface/cytoplasm of most normal cells. AS1411 has shown promising clinical activity and is being widely used as a tumor-targeting agent, but its mechanism of action is not fully understood. Previously, we showed that AS1411 is taken up in cancer cells by macropinocytosis (fluid phase endocytosis) and subsequently stimulates further macropinocytosis by a nucleolin-dependent mechanism. In the current study, we have investigated the significance and molecular mechanisms of AS1411-induced macropinocytosis. Our results indicate that the antiproliferative activity of AS1411 in various cell lines correlated with its capacity to stimulate macropinocytosis. In DU145 prostate cancer cells, AS1411 induced activation of EGFR, Akt, p38, and Rac1. Activation of Akt and p38 were not critical for AS1411 activity because Akt activation was not observed in all AS1411-responsive cell lines and knockdown of p38 had no effect on AS1411's ability to inhibit proliferation. On the other hand, activation of EGFR and Rac1 appeared to play a role in AS1411 activity in all cancer cell lines examined (DU145, MDA-MB-468, A549, LNCaP) and their inhibition significantly reduced AS1411-mediated macropinocytosis and AS1411 antiproliferative activity. Interestingly, downregulation of nucleolin expression by siRNA also produced a substantial increase in activated Rac1, revealing a previously unknown role for nucleolin as a negative regulator of Rac1 activation. Our results are consistent with a model whereby AS1411 binding to nucleolin leads to sustained activation of Rac1 and causes methuosis, a novel type of nonapoptotic cell death characterized by hyperstimulation of macropinocytosis. We speculate that methuosis is a tumor/metastasis suppressor mechanism that opposes the malignant functions of Rac1 and that cancer cells may overexpress nucleolin to surmount this barrier.


Action video games improve reading abilities and visual-to-auditory attentional shifting in English-speaking children with dyslexia.
Dyslexia is characterized by difficulties in learning to read and there is some evidence that action video games (AVG), without any direct phonological or orthographic stimulation, improve reading efficiency in Italian children with dyslexia. However, the cognitive mechanism underlying this improvement and the extent to which the benefits of AVG training would generalize to deep English orthography, remain two critical questions. During reading acquisition, children have to integrate written letters with speech sounds, rapidly shifting their attention from visual to auditory modality. In our study, we tested reading skills and phonological working memory, visuo-spatial attention, auditory, visual and audio-visual stimuli localization, and cross-sensory attentional shifting in two matched groups of English-speaking children with dyslexia before and after they played AVG or non-action video games. The speed of words recognition and phonological decoding increased after playing AVG, but not non-action video games. Furthermore, focused visuo-spatial attention and visual-to-auditory attentional shifting also improved only after AVG training. This unconventional reading remediation program also increased phonological short-term memory and phoneme blending skills. Our report shows that an enhancement of visuo-spatial attention and phonological working memory, and an acceleration of visual-to-auditory attentional shifting can directly translate into better reading in English-speaking children with dyslexia.


Spatial template extraction for image retrieval by region matching.
This paper presents a template and its relation extraction and estimation (TREE) algorithm for indexing images from picture libraries with more semantics-sensitive meanings. This algorithm can learn the commonality of visual concepts from multiple images to give a middle-level understanding about image contents. In this approach, each image is represented by a set of templates and their spatial relations as keys to capture the essence of this image. Each template is characterized by a set of dominant regions, which reflect different appearances of an object at different conditions and can be obtained by the template extraction and analysis (TEA) algorithm through region matching. The spatial template relation extraction and measurement (STREAM) algorithm is then proposed for obtaining the spatial relations between these templates. Due to the nature of a template, which can represent object's appearances at different conditions, the proposed approach owns better capabilities and flexibilities to capture image contents than traditional region-based methods. In addition, through maintaining the spatial layout of images, the semantic meanings of the query images can be extracted and lead to significant improvements in the accuracy of image retrieval. Since no time-consuming optimization process is involved, the proposed method learns the visual concepts extremely fast. Experimental results are provided to prove the superiority of the proposed method.


The effects of microstimulation and microlesions in the ventral and dorsal respiratory groups in medulla of cat.
The responses in respiratory outflow resulting from microstimulation and successive microlesions of the dorsal (DRG) and ventral (VRG) respiratory groups of neurons in the brainstem were studied in anesthetized, paralyzed, artificially ventilated cats. Microstimulation (2 to 120 Hz; 5 to 50 microA; 100 musec pulse duration) at almost every point within the DRG or VRG produced a bilateral short latency inhibition of phrenic nerve activity which had an onset latency of 4 to 9 msec and a duration of 4 to 24 msec. This global stereotyped phrenic inhibition was elicited by single pulses and often was accompanied by a postinhibitory excitation. In 48% (92/193) of the stimulation trials, trains of stimulus pulses during inspiration decreased the duration of inspiration. In 25% of the expiratory microstimulation trials, expiratory duration was increased and in 11%, expiration was shortened markedly by trains of pulses. Single shocks delivered to the right VRG or DRG produced a short latency excitation in the ipsilateral recurrent laryngeal nerve (RRL). This RRL excitation had an onset latency of 2 to 5 msec and a duration of 3 to 15 msec. Evidence suggests that the RRL excitation is due to a paucisynaptic activation of expiratory motoneurons in the caudal VRG. This activation is synchronous with the inhibition of inspiratory neurons in DRG and VRG. Despite the powerful short latency effects of microstimulation in VRG and DRG, extensive bilateral destruction of these neuronal populations had only modest effects on respiratory rhythm, while it decreased (or abolished) respiratory outflow in phrenic and recurrent laryngeal nerves. The combined results of the microstimulation and microlesion portions of this study suggest that a region (or regions) outside of the DRG and VRG might be important in the control of the respiratory pattern and that the DRG and VRG are important in determining the depth of inspiration; their role in generating respiratory rhythm needs to be critically re-examined.


Classifying Vocal Folds Fixation from Endoscopic Videos with Machine Learning.
Vocal folds motility evaluation is paramount in both the assessment of functional deficits and in the accurate staging of neoplastic disease of the glottis. Diagnostic endoscopy, and in particular videoendoscopy, is nowadays the method through which the motility is estimated. The clinical diagnosis, however, relies on the examination of the videoendoscopic frames, which is a subjective and professional-dependent task. Hence, a more rigorous, objective, reliable, and repeatable method is needed. To support clinicians, this paper proposes a machine learning (ML) approach for vocal cords motility classification. From the endoscopic videos of 186 patients with both vocal cords preserved motility and fixation, a dataset of 558 images relative to the two classes was extracted. Successively, a number of features was retrieved from the images and used to train and test four well-grounded ML classifiers. From test results, the best performance was achieved using XGBoost, with precision = 0.82, recall = 0.82, F1 score = 0.82, and accuracy = 0.82. After comparing the most relevant ML models, we believe that this approach could provide precise and reliable support to clinical evaluation.Clinical Relevance- This research represents an important advancement in the state-of-the-art of computer-assisted otolaryngology, to develop an effective tool for motility assessment in the clinical practice.


[Efficacy and Survival of Venetoclax Based Regimen in the Treatment of Acute Myeloid Leukemia].
To explore the efficacy and survival of venetoclax based (VEN-based) regimen in the treatment of acute myeloid leukemia(AML). A retrospective study was conducted in patients who received VEN-based regimen and completed at least 1 course of efficacy evaluation at the The First Affiliated Hospital of Nanchang University from July 2019 to July 2022. The incidence of complete remission (CR)/CR with incomplete hematologic recovery (CRi) rate, objective remission rate(ORR) and survival of patients with different risk strati- fication and gene subtypes were analyzed. A total of 79 patients were enrolled, including 43 patients with newly diagnosed unfit AML (unfit AML) and 36 relapsed/refractory AML (R/R AML). The median age of the patients was 62(14-83) years old. 36 out of 79 patients achieved CR/CRi and the ORR of the whole cohort was 64.6%. The CR/CRi rate of unfit AML patients was significantly higher than that of R/R AML patients (60.5% vs 27.8%, P=0.004). In unfit AML cohort, the patients with NPM1 and IDH1/2 mutations were benefited, 8 out of 9 patients ahcieved CR/CRi, 7/8 and 5/8 patients achieved minimal residual disease (MRD) negativity, respectively. Six out of 9 patients with TET2 mutation achieved CR/CRi, 3/6 patients achieved MRD negativity. In R/R AML cohort, 2 out of 3 patients with RUNX1 mutation achieved CR/CRi, without MRD negative, while the CR/CRi rate of patients with other gene mutations was lower than 40%. The median follow-up time was 10.1(95%CI: 8.6-11.6) months. In whole cohort, the median overall survival (mOS) time was 9.1 months and the relapse free survival (RFS) time was not reached. The mOS and RFS of unfit AML patients were significantly longer than those of R/R AML patients (14.1 vs 6.8 months, P=0.013; not reached vs 3.3 months, P=0.000). In unfit AML cohort, the mOS of patients with NPM1 or IDH1/2 mutations was not reached, while that of patients without NPM1 or IDH1/2 mutations was 8.0 months (P=0.009; P=0.022). Furthermore, the mOS of patients with TP53 mutaion was significantly shorter than that of patients without TP53 mutation (5.2 vs 14.1 months, P=0.049). In R/R AML cohort, there was no significant difference in mOS between patients with mutation in each gene subtype and those without gene mutation (P>0.05). All patients had hematology adverse reactions, 91.1% patients had AE grade≥3. The most common non-hematology adverse reactions was infection, with an incidence of 91.1%. VEN-based regimen was tolerable for AML patients. VEN-based regimen can achieve a high response rate, especially in unfit AML with acceptable safety, and some patients can achieve MRD negative. It is also effective in NPM1-, IDH1/2-positive patients with long survival time.


Integrated modeling of extended agro-food supply chains: A systems approach.
The current intense food production-consumption is one of the main sources of environmental pollution and contributes to anthropogenic greenhouse gas emissions. Organic farming is a potential way to reduce environmental impacts by excluding synthetic pesticides and fertilizers from the process. Despite ecological benefits, it is unlikely that conversion to organic can be financially viable for farmers, without additional support and incentives from consumers. This study models the interplay between consumer preferences and socio-environmental issues related to agriculture and food production. We operationalize the novel concept of extended agro-food supply chain and simulate adaptive behavior of farmers, food processors, retailers, and customers. Not only the operational factors (e.g., price, quantity, and lead time), but also the behavioral factors (e.g., attitude, perceived control, social norms, habits, and personal goals) of the food suppliers and consumers are considered in order to foster organic farming. We propose an integrated approach combining agent-based, discrete-event, and system dynamics modeling for a case of wine supply chain. Findings demonstrate the feasibility and superiority of the proposed model over the traditional sustainable supply chain models in incorporating the feedback between consumers and producers and analyzing management scenarios that can urge farmers to expand organic agriculture. Results further indicate that demand-side participation in transition pathways towards sustainable agriculture can become a time-consuming effort if not accompanied by the middle actors between consumers and farmers. In practice, our proposed model may serve as a decision-support tool to guide evidence-based policymaking in the food and agriculture sector.


[Spectral analytical study on the developmental properties of EEG in children: Generalized and localized components of EEG spectra in the normal and the mentally retarded children (author's transl)].
The purpose of the present study was to elucidate the developmental properties of EEG in the normal and the mentally retarded children through regional differences and regional regional interrelationships. The localized components appearing at 1 or 2 regions tended to show low coherences in relation to the occipital region. The generalized component, observed over all regions, indicated lower coherences in the theta component than in the alpha one. It may be inferred that the theta component shown in the developing children may be differenciated from the component around 10 Hz.


Hepatocyte nuclear factor 4 alpha polymorphisms and the metabolic syndrome in French-Canadian youth.
Hepatocyte nuclear factor 4 alpha (HNF4α) is a transcription factor involved in the regulation of serum glucose and lipid levels. Several HNF4A gene variants have been associated with the risk of developing type 2 diabetes mellitus. However, no study has yet explored its association with insulin resistance and the cardiometabolic risk in children. We aimed to investigate the relationship between HNF4A genetic variants and the presence of metabolic syndrome (MetS) and metabolic parameters in a pediatric population. Our study included 1,749 French-Canadians aged 9, 13 and 16 years and evaluated 24 HNF4A polymorphisms that were previously identified by sequencing. Analyses revealed that, after correction for multiple testing, one SNP (rs736824; P<0.022) and two haplotypes (P1 promoter haplotype rs6130608-rs2425637; P<0.032 and intronic haplotype rs736824-rs745975-rs3212183; P<0.025) were associated with the risk of MetS. Additionally, a significant association was found between rs3212172 and apolipoprotein B levels (coefficient: -0.14 ± 0.05; P<0.022). These polymorphisms are located in HNF4A P1 promoter or in intronic regions. Our study demonstrates that HNF4α genetic variants are associated with the MetS and metabolic parameters in French Canadian children and adolescents. This study, the first exploring the relation between HNF4A genetic variants and MetS and metabolic variables in a pediatric cohort, suggests that HNF4α could represent an early marker for the risk of developing type 2 diabetes mellitus.


GR, Sgk1, and NDRG1 in esophageal squamous cell carcinoma: their correlation with therapeutic outcome of neoadjuvant chemotherapy.
Esophageal squamous cell carcinoma (ESCC) is a highly malignant neoplasm. The glucocorticoid (GC)-glucocorticoid receptor (GR) pathway plays pivotal roles in cellular response to various stresses of tumor cells, including chemotherapy. However, the status of the GC-GR pathway in ESCC, including its correlation with chemotherapeutic responses, is largely unknown. GR, serum-and glucocorticoid-regulated kinase 1 (Sgk1), and N-myc down regulation gene 1 (NDRG1) were immunolocalized in 98 patients with ESCC who had undergone esophagectomy following neoadjuvant chemotherapy (NAC) with 2 courses of 5-fluorouracil + cisplatin. We also examined biopsy specimens before NAC in 42 cases and compared the results between those before and after NAC. Overall survival (OS) of the patients treated with surgery following NAC was significantly shorter in the group with high GR than that with low GR status (P = 0.0473). Both OS and disease-free survival (DFS) were significantly shorter in both Sgk1- and NDRG1-high groups than in the low groups (OS: Sgk1, P = 0.0055; NDRG1, P = 0.0021; DFS: Sgk1, P = 0.0240; NDRG1, P = 0.0086). Biopsy specimens before NAC showed significantly shorter DFS in the high Sgk1 group (P = 0.0095), while both OS and DFS were shorter in the high NDRG1 group (OS, P = 0.0233; DFS, P = 0.0006) than in the respective low groups. In the high NDRG1 group of biopsy specimens before NAC, the tumor reduction rate by NAC was significantly attenuated (P = 0.021). High GR, Sgk1, and NDRG1 statuses in ESCC after NAC was significantly associated with an overall worse prognosis, with no significant changes in their expression levels before and after NAC. Therefore, increased activity of the GC-GR pathway with enhanced induction of Sgk1 and NDRG1 in carcinoma cells play pivotal roles in tumor progression and development of chemo-resistance in patients with ESCC undergoing NAC.


Diclofenac-Choline Antioxidant Activity Investigated by means of Luminol Amplified Chemiluminescence of Human Neutrophil Bursts and Electron Paramagnetic Resonance Spectroscopy.
A new diclofenac salt called diclofenac-choline (DC) has recently been proposed for the symptomatic treatment of oropharyngeal inflammatory processes and pain because its greater water solubility allows the use of high concentrations, which are useful when the contact time between the drug and the oropharyngeal mucosa is brief, as in the case of mouthwashes or spray formulations. The antioxidant activity of DC has not yet been investigated, and so the aim was to use luminol-amplified-chemiluminescence (LACL) to verify whether various concentrations of DC (1.48, 0.74 and 0.37 mg/mL for incubation times of 2, 4 and 8 min) interfere with oxygen and nitrogen radicals during the course of human neutrophils respiratory bursts; electron paramagnetic resonance (EPR) spectroscopy was used to investigate its direct antiradical (scavenger) activity. The EPR findings showed that DC has concentration-dependent scavenging activity against the ABTS, the DPPH, and the hydroxyl radicals, but no activity on superoxide anion, as has been previously reported in the case of other NSAIDs. LACL revealed an inhibitory effect that was statistically significant after only 2 min of incubation, and similar after 4 and 8 min. The effects on the peroxynitrite radical paralleled those observed in the previous test. High concentrations and short incubation times showed that there is no interference on PMN viability, and so the inhibitory findings must be attributed to the effect of the drug. The anti-inflammatory effects of DC cannot be attributed solely to the inhibition of prostaglandin synthesis, but its effects on free radicals and neutrophil bursts suggest that they may contribute to its final therapeutic effect.


Mild transcobalamin I (haptocorrin) deficiency and low serum cobalamin concentrations.
Low cobalamin concentrations are common, but their causes are often unknown. Transcobalamin I/haptocorrin (TC I/HC) deficiency, viewed as a rare cause, has not been examined systematically in patients with unexplained low serum cobalamin. Total TC I/HC was measured by RIA in three subgroups of 367, 160, and 38 patients with different categories of low cobalamin concentrations and three comparison subgroups of 112, 281, and 119 individuals with cobalamin concentrations within the reference interval. Additional studies, including family studies, were done in selected patients found to have low TC I/HC concentrations. Low TC I/HC concentrations suggestive of mild TC I/HC deficiency occurred in 54 of 367 (15%) patients with low cobalamin identified by clinical laboratories and 24 of 160 (15%) patients whose low cobalamin was unexplained after absorption and metabolic evaluation, but in only 2 of 38 patients with malabsorptive causes of low cobalamin concentrations (5%). The prevalence was only 3% (8 of 281 plasma samples) to 5% (6 of 112 sera) in patients with cobalamin concentrations within the reference interval and 3% (4 of 119) in healthy volunteers. Three patients with low cobalamin (0.6%) had severe TC I/HC deficiency with undetectable TC I/HC. Presumptive heterozygotes for severe TC I/HC deficiency in two families had the findings of mild TC I/HC deficiency; mild deficiency was also found in at least three of seven studied families of patients with mild TC I/HC deficiency. Mild TC I/HC deficiency is frequently associated with low cobalamin, is often familial, and its biochemical phenotype appears identical to the heterozygous state of severe TC I/HC deficiency. Severe TC I/HC deficiency also appears to be more common than suspected. Both diagnoses should be considered in all patients with unexplained low serum cobalamin.


Microbial communities acclimate to recurring changes in soil redox potential status.
Rapidly fluctuating environmental conditions can significantly stress organisms, particularly when fluctuations cross thresholds of normal physiological tolerance. Redox potential fluctuations are common in humid tropical soils, and microbial community acclimation or avoidance strategies for survival will in turn shape microbial community diversity and biogeochemistry. To assess the extent to which indigenous bacterial and archaeal communities are adapted to changing in redox potential, soils were incubated under static anoxic, static oxic or fluctuating redox potential conditions, and the standing (DNA-based) and active (RNA-based) communities and biogeochemistry were determined. Fluctuating redox potential conditions permitted simultaneous CO₂ respiration, methanogenesis, N₂O production and iron reduction. Exposure to static anaerobic conditions significantly changed community composition, while 4-day redox potential fluctuations did not. Using RNA:DNA ratios as a measure of activity, 285 taxa were more active under fluctuating than static conditions, compared with three taxa that were more active under static compared with fluctuating conditions. These data suggest an indigenous microbial community adapted to fluctuating redox potential.


DISASTER MEDICAL CARE AND SHELTER-The Federal Program.
The role of the physician in event of natural disaster or overwhelming (perhaps nuclear) attack by an enemy is:To assist the layman in preparing to meet his own health needs in a disaster situation until organized health services can reach him. To prepare and plan for the provision of organized medical care when conditions permit. To extend his own capability to render medical care outside his normal specialty. To assist in the training of allied and professional health workers and laymen for specific mobilization assignments in health services.


Betamethasone-mediated activation of biphenyl 2-hydroxylation in rat liver microsomes. Studies on possible mechanisms.
The addition of the steroid betamethasone to intact or detergent-solubilized rat liver microsomes caused a concentration-dependent increase in the rate of biphenyl 2-hydroxylation. Betamethasone (100 microM) increased the apparent Vmax for 2-hydroxybiphenyl formation 2- to 4-fold but had no effect on the apparent Km when either the biphenyl or NADPH concentration was varied. Betamethasone had little or no effect on the apparent Vmax or apparent Km of the 3- and 4-hydroxylations of biphenyl. The steroid did not enhance biphenyl 2-hydroxylation through a peroxidative mechanism. Betamethasone had little or no effect on the rate of the NADPH-dependent reduction of cytochrome c or total microsomal cytochrome P-450. The addition of purified NADPH-cytochrome P-450 reductase to cholate-solubilized liver microsomes increased the rate of hydroxylation of biphenyl in positions 2 and 4. Betamethasone (100 microM) decreased the apparent Km for purified cytochrome P-450 reductase by 48% and increased the apparent Vmax of 2-hydroxybiphenyl formation by 2-fold when the concentration of cytochrome P-450 reductase was varied. The steroid did not alter the Km or Vmax values for the 4-hydroxylation of biphenyl. The data suggest that betamethasone enhances the interaction between the reductase and the form(s) of cytochrome P-450 responsible for the 2-hydroxylation of biphenyl.


Basolateral amygdala cdk5 activity mediates consolidation and reconsolidation of memories for cocaine cues.
Cocaine use and relapse involves learned associations between cocaine-associated environmental contexts and discrete stimuli and cocaine effects. Initially, these contextual and discrete cues undergo memory consolidation after being paired with cocaine exposure. During abstinence, cocaine cue memories can undergo memory reconsolidation after cue exposure without the drug. We used a conditioned place preference (CPP) procedure in rats to study the role of neuronal protein kinase cyclin-dependent kinase 5 (Cdk5) in consolidation and reconsolidation of cocaine cue memories. We found that the expression of cocaine CPP in drug-free tests 1 d after CPP training (four pairings of 10 mg/kg cocaine with one context and four pairings of saline with a different context) increased Cdk5 activity, and levels of the Cdk5 activator p35 in basolateral but not central amygdala. We also found that basolateral (but not central) amygdala injections of the Cdk5 inhibitor beta-butyrolactone (100 ng/side) immediately (but not 6 h) after cocaine-context pairings during training prevented subsequent cocaine CPP expression. After training, acute basolateral (but not central) amygdala beta-butyrolactone injections immediately before testing prevented the expression of cocaine CPP; this effect was also observed on a second test performed 1 d later, suggesting an effect on reconsolidation of cocaine cue memories. In support, basolateral beta-butyrolactone injections, given immediately (but not 6 h) after a single exposure to the cocaine-paired context, prevented cocaine CPP expression 1 and 14 d after the injections. Results indicate that basolateral amygdala Cdk5 activity is critical for consolidation and reconsolidation of the memories of cocaine-associated environmental cues.


Safety of valproic acid in patients with chronic obstructive pulmonary disease: a population-based cohort study.
Valproic acid is an anticonvulsant that also inhibits histone deacetylase (HDAC), a property that could worsen pulmonary function in patients with chronic obstructive pulmonary disease (COPD). The clinical significance of this property is unknown. We therefore compared the risk of COPD exacerbation in older patients with COPD commencing treatment with either valproic acid or phenytoin, an anticonvulsant that does not affect HDAC. We conducted a population-based retrospective cohort study of Ontario residents with COPD aged 66 years or older who started treatment with valproic acid or phenytoin between 1 April 1993 and 30 November 2012. The primary outcome was a hospital admission or emergency department visit for a COPD exacerbation within 240 days of drug initiation. A secondary outcome examined initiation of oral corticosteroids in the outpatient setting. During the study period, we identified 4596 COPD patients who commenced valproic acid and 8478 who commenced phenytoin. Following multivariable adjustment, valproic acid did not increase the risk of the primary outcome (adjusted hazard ratio 1.00, 95% confidence interval 0.79 to 1.26). Although valproic acid was associated with a lower risk of initiating oral corticosteroids in the first thirty days following commencement of anticonvulsant therapy (adjusted hazard ratio 0.32; 95% confidence interval 0.21 to 0.49), no difference was observed during subsequent follow-up. Among older patients with COPD, treatment with valproic acid does not increase the risk of adverse pulmonary outcomes relative to phenytoin. These findings suggest that valproate-induced HDAC inhibition is of little clinical relevance in this context.


Characterization of a cryptic plasmid from Pseudomonas sp. and utilization of its temperature-sensitive derivatives for genetic manipulation.
A new cryptic plasmid, named pMM101, in an environmental Pseudomonas sp. has been isolated, and its 2140-bp DNA sequence has been determined and analyzed. One open reading frame (Rep(MM)) similar to the rep gene of the rolling-circle replicon (RCR) group VIII has been allocated in the replication region where a putative double-strand origin dso and single-strand origin sso could also be identified. After in vitro mutagenesis by error-prone PCR and introduced into Pseudomonas aeruginosa PAO1, a mutant plasmid which was stably maintained at 30 degrees C but not at 42 degrees C was isolated and analyzed. It had two nucleotide substitutions in the putative sso and dso regions. Replacing one of the two sites with wild type sequence resulted in similar instability at 42 degrees C, indicating either mutation could result in the temperature-sensitive trait. Using the mutant replicon, a Pseudomonas (temperature-sensitive)-Escherichia coli shuttle vector, pUM109, has been constructed and used successfully to disrupt P. aeruginosa genes. This temperature-sensitive vector provides an alternative tool for genetic manipulations in this industrially as well as medically important bacterium.


SARS-CoV-2 Receptor Angiotensin I-Converting Enzyme Type 2 (ACE2) Is Expressed in Human Pancreatic β-Cells and in the Human Pancreas Microvasculature.
Increasing evidence demonstrated that the expression of Angiotensin I-Converting Enzyme type 2 (ACE2) is a necessary step for SARS-CoV-2 infection permissiveness. In light of the recent data highlighting an association between COVID-19 and diabetes, a detailed analysis aimed at evaluating ACE2 expression pattern distribution in human pancreas is still lacking. Here, we took advantage of INNODIA network EUnPOD biobank collection to thoroughly analyze ACE2, both at mRNA and protein level, in multiple human pancreatic tissues and using several methodologies. Using multiple reagents and antibodies, we showed that ACE2 is expressed in human pancreatic islets, where it is preferentially expressed in subsets of insulin producing β-cells. ACE2 is also highly expressed in pancreas microvasculature pericytes and moderately expressed in rare scattered ductal cells. By using different ACE2 antibodies we showed that a recently described short-ACE2 isoform is also prevalently expressed in human β-cells. Finally, using RT-qPCR, RNA-seq and High-Content imaging screening analysis, we demonstrated that pro-inflammatory cytokines, but not palmitate, increase ACE2 expression in the β-cell line EndoC-βH1 and in primary human pancreatic islets. Taken together, our data indicate a potential link between SARS-CoV-2 and diabetes through putative infection of pancreatic microvasculature and/or ductal cells and/or through direct β-cell virus tropism.


Comparative sequence analysis of human cytomegalovirus strains.
Three regions of DNA from five low-passage clinical isolates of human cytomegalovirus were amplified by polymerase chain reaction. The DNA sequences as well as the predicted amino acid sequences were compared with those of the laboratory strains AD169 and Towne. The genomic regions consisted of (i) three regions from the major glycoprotein (gp58/116, unique long [UL]55), (ii) three regions from the integral membrane protein (IMP, UL100), and (iii) a region from the major immediate-early 1 and 2 (IE-1/2) enhancer/promoter. Homologies ranged from 75.8 to 100.0% on the nucleotide level and from 47 to 100% on the amino acid level. The following two patterns were observed. (i) There are regions with a high degree of conservation with few scattered point mutations (mainly in the IE-1/2 enhancer/promoter and in the IMP gene). (ii) There are clusters of highly variable regions (parts of the gp58/116 gene and of the IMP gene). Within the areas of high variability, the strains could be classified into a limited number of subtypes.


Isolation of two novel WNT genes, WNT14 and WNT15, one of which (WNT15) is closely linked to WNT3 on human chromosome 17q21.
The Wnt gene family consists of at least 15 structurally related genes that encode secreted extracellular signaling factors. Wnt proteins function in a range of critical developmental processes in both vertebrates and invertebrates and are implicated in regulation of cell growth and differentiation in certain adult mammalian tissues, including the mammary gland. We have isolated a number of WNT sequences from human genomic DNA, two of which, designated WNT14 and WNT15, represent novel members of the Wnt gene family. We also isolated WNT sequences from human mammary cDNA and present evidence that WNT13 is expressed in human breast tissue, in addition to those previously described. WNT14 and WNT15 appear to have originated from an ancestral branch of the Wnt gene family that also includes the Wnt9 sequences found in jawless and cartilaginous fishes. A Wnt14 cDNA was also isolated from chicken and a partial Wnt15 sequence from mouse. We show that human WNT14 maps to chromosome 1 and that WNT15 maps distal to BRCA1 on chromosome 17q21, where it lies within 125 kb of another WNT family member, WNT3.


Epidemiology of rheumatic heart disease in black shcoolchildren of Soweto, Johannesburg.
A survey to determine the prevalence of rheumatic heart disease (R.H.D.) in Black children was conducted in the creeches and primary schools of the South Western Townships of Johannesburg (Soweto). A total of 12 050 Black children were examined by 10 cardiologists in May to October 1972. The overal prevalence rate of R.H.D. was 6.9 per 1000, with a peak rate of 19.2 per 1000 in children of the seventh school grade. The maximal age incidence was 15-18 years and there was a female preponderance of 1 6:1. A rise in prevalence occurred with increasing family size. Most children (92%) were asymptomatic, and in 82.5% R.H.D. was diagnosed for the first time during the school survey. The commonest valve lesion was mitral regurgitation, which was present in 93% and occurred as an isolated lesion in 47.5%. Lancefield's group A beta-haemolytic streptococcus was isolated from the throats of 52 per 1000 Soweto children. The auscultatory features of a non-ejection systolic click and late systolic murmur were prevalent (13.9 per 1000) and had several epidemiological factors in common with R.H.D. A comprehensive preventative campaign is urgently needed in South Africa, directed at both primary and secondary prophylaxis of R.H.D. The socioeconomic status of the community must be improved if optimal prevention is to be achieved.


Overexpression of the p21 sdi1 gene induces senescence-like state in human cancer cells: implication for senescence-directed molecular therapy for cancer.
Normal cells in a culture enter a nondividing state after a finite number of population doubling, which is termed replicative senescence, whereas cancer cells have unlimited proliferative potential and are thought to exhibit an immmortal phenotype by escaping from senescence. The p21 gene (also known as sdi1), which encodes the cyclin-dependent kinase inhibitor, is expressed at high levels in senescent cells and contributes to the growth arrest. To examine if the p21sdi1 gene transfer could induce senescence in human cancer cells, we utilized an adenoviral vector-based expression system and four human cancer cell lines differing in their p53 status. Transient overexpression of p21sdi1 on cancer cells induced quiescence by arresting the cell cycle at the G1 phase and exhibited morphological changes, such as enlarged nuclei as well as a flattened cellular shape, specific to the senescence phenotype. We also showed that p21sdi1-transduced cancer cells expressed beta-galactosidase activity at pH 6.0, which is known to be a marker of senescence. Moreover, the polymerase chain reaction-based assay demonstrated that levels of telomerase activity were significantly lower in p21sdi1-expressing cells compared to parental cancer cells. These observations provide the evidence that p21sdi1 overexpression could induce a senescence-like state and reduce telomerase activity in human cancer cells, suggesting that these novel p21sdi1 functions may have important implications for anticancer therapy.


Genetic architecture of differences in oviposition preference between ancestral and derived populations of the seed beetle Acanthoscelides obtectus.
We investigated the additive, dominance and epistatic genetic effects underlying differentiation in oviposition preference between two populations of the seed beetle Acanthoscelides obtectus evolved in the laboratory for 102 generations on bean and chickpea seeds. We reared and tested females on each of two host legumes. The populations differed in mean oviposition preference; the preference for chickpea was stronger in population reared on the chickpea (C) than in population maintained on common bean (P). Observations in the parental populations indicated that females tend to prefer ovipositioning their eggs on the seeds they have already experienced. The patterns of the means in each of the parental populations and 12 types of hybrids (two F(1), two F(2) and eight backcrosses) indicated that population differences in oviposition preference from both rearing hosts could be explained by nonadditive genetic effects. Statistically detectable additive and dominance genetic effects were observed in the most parsimonious model only when females were reared on the chickpea. The most parsimonious models on both rearing hosts suggested a contribution of negative additive x additive epistasis to the divergence of oviposition preference between the P and C populations. This indicates a positive effect of epistasis on the performance of the second generations of hybrids.


Immunocytochemical detection of the 70-kd heat shock protein in alcoholic liver disease.
Exposure of cells to physical (eg, heat) or chemical (eg, alcohol) stress results in increased synthesis of a set of highly conserved polypeptides termed heat shock proteins (HSPs), among which the 70-kd protein (HSP 70) is one of the most consistently inducible and highly conserved. This HSP has adenosine triphosphate-binding properties and is known to associate strongly with cytoskeletal structures that are usually disrupted on injury by heat or alcohol. Some HSPs apparently function as accessories to a nonlysosomal, adenosine triphosphate-dependent proteolytic system that binds and digests away stress-generated abnormal or denatured proteins after their conjugation with ubiquitin, a small HSP. Ubiquitin has been demonstrated immunocytochemically in Mallory bodies, which represent mainly degenerated intermediate filaments accumulated in hepatocytes of alcoholic-diseased liver. We immunostained histologic sections from patients with alcoholic liver disease using a polyclonal antibody raised against HSP 70. Strong diffuse cytoplasmic immunoreactivity was observed in many hepatocytes, including cells without Mallory bodies or fatty degeneration. Positive immunoreactivity for HSP 70 points to a possible involvement of this HSP in the pathogenesis of alcoholic liver disease. It also suggests that immunocytochemical detection of HSP 70 may serve as a more sensitive indicator of hepatocellular injury.


Municipal solid-waste management in Istanbul.
Istanbul, with a population of around 13 million people, is located between Europe and Asia and is the biggest city in Turkey. Metropolitan Istanbul produces about 14,000 tons of solid waste per day. The aim of this study was to assess the situation of municipal solid-waste (MSW) management in Istanbul. This was achieved by reviewing the quantity and composition of waste produced in Istanbul. Current requirements and challenges in relation to the optimization of Istanbul's MSW collection and management system are also discussed, and several suggestions for solving the problems identified are presented. The recovery of solid waste from the landfills, as well as the amounts of landfill-generated biogas and electricity, were evaluated. In recent years, MSW management in Istanbul has improved because of strong governance and institutional involvement. However, efforts directed toward applied research are still required to enable better waste management. These efforts will greatly support decision making on the part of municipal authorities. There remains a great need to reduce the volume of MSW in Istanbul.


The D-lightful vitamin D for child health.
Vitamin D deficiency and insufficiency is common in children. Vitamin D deficiency is defined as a 25-hydroxyvitamin D <20 ng/mL and insufficiency as 21-29 ng/mL. The major sources of vitamin D are sun exposure, diet, and supplements. In young children, vitamin D deficiency causes a mineralization defect of the skeleton, resulting in rickets. Vitamin D deficiency has been linked to many chronic diseases, including multiple sclerosis, type 1 diabetes, infectious diseases, heart disease, and type 2 diabetes. In utero vitamin D deficiency increases risk for preeclampsia and the need for a cesarean section. Children living at higher latitudes and who are presumed to be at increased risk for vitamin D deficiency are at higher risk for developing type 1 diabetes and multiple sclerosis later in life. The Institute of Medicine recommends that children 0-1 year and 1-18 years need 400 and 600 IU/d, respectively, of vitamin D for maximal skeletal health. The Endocrine Society's Clinical Guidelines recommended that to prevent and treat vitamin D deficiency, children 0-1 year and 1-18 years need 400-1000 and 600-1000 IU/d, respectively.


Global analysis of human nonreceptor tyrosine kinase specificity using high-density peptide microarrays.
Protein kinases phosphorylate substrates in the context of specific phosphorylation site sequence motifs. The knowledge of the specific sequences that are recognized by kinases is useful for mapping sites of phosphorylation in protein substrates and facilitates the generation of model substrates to monitor kinase activity. Here, we have adapted a positional scanning peptide library method to a microarray format that is suitable for the rapid determination of phosphorylation site motifs for tyrosine kinases. Peptide mixtures were immobilized on glass slides through a layer of a tyrosine-free Y33F mutant avidin to facilitate the analysis of phosphorylation by radiolabel assay. A microarray analysis provided qualitatively similar results in comparison with the solution phase peptide library "macroarray" method. However, much smaller quantities of kinases were required to phosphorylate peptides on the microarrays, which thus enabled a proteome scale analysis of kinase specificity. We illustrated this capability by microarray profiling more than 80% of the human nonreceptor tyrosine kinases (NRTKs). Microarray results were used to generate a universal NRTK substrate set of 11 consensus peptides for in vitro kinase assays. Several substrates were highly specific for their cognate kinases, which should facilitate their incorporation into kinase-selective biosensors.


Seroprevalence of Toxoplasma gondii in farm-reared ostriches and wild game species from Zimbabwe.
One hundred and seventy one serum samples from 10 game species from Zimbabwe were tested for IgG antibodies to Toxoplasma gondii infection using the modified agglutination test (MAT). Significantly higher seroprevalences were found in the felidae (Panthera leo) (92% of 26), bovidae (Tragelaphus species) (55.9% of 34) and farm-reared struthionidae (Struthio camelus) (48% of 50) compared to the other groups tested. Among the bovidae, the nyala (Tragelaphus angasii) had the highest seroprevalence of 90% (9/10). Anti-Toxoplasma antibody prevalences in browsers [greater kudu (Tragelaphus strepsiceros) (20% of 10), giraffe (Giraffa camelopardalis) (10% of 10) and elephant (Loxodonta africana) (10% of 20)] were generally in the lower range. No antibodies were detected in the wild African suidae [warthog (Phacochoerus africanus) and bushpig (Potamochoerus larvatus)]. Attempts to isolate T. gondii from the heart muscles of seropositve ostriches by subinoculation in BALB/c mice were unsuccessful.


Protective Effect of Lactobacillus rhamnosus GG and its Supernatant against Myocardial Dysfunction in Obese Mice Exposed to Intermittent Hypoxia is Associated with the Activation of Nrf2 Pathway.
Prolonged intermittent hypoxia (IH) has been shown to impair myocardial function (mainly via oxidative stress and inflammation) and modify gut microbiota in mice. Gut microbiota plays an important role in health and disease, including obesity and cardiovascular disease (CVD). Probiotics refer to live microorganisms that confer health benefits on the host after administration in adequate amounts. Research on novel probiotics related therapies has evoked much attention. In our previous study, both Lactobacillus rhamnosus GG (LGG) and LGG cell-free supernatant (LGGs) were found to protect against alcohol-induced liver injury and steatosis; however, the effects of LGG and LGGs on cardiac tissues of obese mice exposed to IH have not been determined. Here we exposed high-fat high-fructose diet (HFHFD)-induced obese mice to IH, to establish a model of obesity with obstructive sleep apnea (OSA). Mice were divided into four groups: (1) HFHFD for 15 weeks; (2) HFHFD for 15 weeks with IH in the last 12 weeks (HFHFD/IH); (3) and (4) HFHFD/IH plus oral administration of either LGG (109 CFU bacteria/day) or LGGs (dose equivalent to 109 CFU bacteria/day) over the 15 weeks, respectively. Compared to HFHFD mice, HFHFD/IH-mice showed heart dysfunction with significant cardiac remodeling and inflammation; all these pathological and functional alterations were prevented by treatment with both LGG and LGGs (no significant difference between LGG and LGGs in this respect). The cardioprotective effect of LGG and LGGs against IH/HFHFD was associated with up-regulation of nuclear factor erythroid 2-related factor 2(Nrf2)-mediated antioxidant pathways. Our findings suggest a cardioprotective effect of LGG and LGGs in obese mice with OSA.


Molecular identification of T cells that respond in a primary bulk culture to a peptide derived from a platelet glycoprotein implicated in neonatal alloimmune thrombocytopenia.
Neonatal alloimmune thrombocytopenia induced by the human platelet alloantigen 1a (HPA1a) is characterized by generation of alloantibodies by a mother who is homozygous for the HPA1b alloantigen and almost always HLA-DRB3*0101. The disease is viewed as B cell mediated but the linkage with HLA is indicative of a role for T cells. The HPA1a and HPA1b allotypes are defined, respectively, by Leu and Pro at amino acid 33 of the beta-chain of the platelet integrin GPIIbIIIa (alpha(IIb)beta3). Under the assumption that the same polymorphism may control both the B cell epitope and constitute the MHC-bound peptide, we restimulated PBMC from a woman with an affected child with a synthetic peptide from this polymorphic region. Molecular analysis of the responding T cell repertoire identified two T cells which predominated in cultures stimulated with the alloantigen peptide and which were absent in cultures with the autoantigen peptide. In spite of the use of different V families, sequence of the CDR3 region of the T cell receptor (TCR) beta-chain revealed the presence of a shared motif, L-P-S/T. Oligonucleotide probes specific for the CDR3 sequence indicated that these T cells were present in the PBMC at the highest levels immediately after delivery of the affected infant and their frequency dropped at later times.


Objective assessment of image quality. III. ROC metrics, ideal observers, and likelihood-generating functions.
We continue the theme of previous papers [J. Opt. Soc. Am. A 7, 1266 (1990); 12, 834 (1995)] on objective (task-based) assessment of image quality. We concentrate on signal-detection tasks and figures of merit related to the ROC (receiver operating characteristic) curve. Many different expressions for the area under an ROC curve (AUC) are derived for an arbitrary discriminant function, with different assumptions on what information about the discriminant function is available. In particular, it is shown that AUC can be expressed by a principal-value integral that involves the characteristic functions of the discriminant. Then the discussion is specialized to the ideal observer, defined as one who uses the likelihood ratio (or some monotonic transformation of it, such as its logarithm) as the discriminant function. The properties of the ideal observer are examined from first principles. Several strong constraints on the moments of the likelihood ratio or the log likelihood are derived, and it is shown that the probability density functions for these test statistics are intimately related. In particular, some surprising results are presented for the case in which the log likelihood is normally distributed under one hypothesis. To unify these considerations, a new quantity called the likelihood-generating function is defined. It is shown that all moments of both the likelihood and the log likelihood under both hypotheses can be derived from this one function. Moreover, the AUC can be expressed, to an excellent approximation, in terms of the likelihood-generating function evaluated at the origin. This expression is the leading term in an asymptotic expansion of the AUC; it is exact whenever the likelihood-generating function behaves linearly near the origin. It is also shown that the likelihood-generating function at the origin sets a lower bound on the AUC in all cases.