An ultrastructural examination of Penicillium urticae during the transition to antibiotic biosynthesis.
The ultrastructure of Penicillium urticae mycelium was compared at various stages of submerged growth to examine changes associated with the onset of antibiotic biosynthesis. Penicillium urticae was shown to be a normal eukaryotic, septate, filamentous fungus with a variety of subcellular components. Younger mycelia possessed a denser cytoplasm which gave way to a more granular and vacuolated cytoplasm as the organism made the transition into antibiotic biosynthesis. An increase in the thickness, and perhaps the structural complexity, of the cell wall also occurred over the transition. There was evidence of a glycocalyx surrounding the hyphae. Discrete granules, termed peripheral particles, appeared and increased in number over the transition. Their biochemical content and possible involvement in patulin production was tested by examining P. urticae after growth in media of different composition, and by examining the ultrastructure of a patulin minus mutant, P3. The significance of these observations in relation to patulin production is discussed.


Deterministic assembly of linear gold nanorod chains as a platform for nanoscale applications.
We demonstrate a method to assemble gold nanorods highly deterministically into a chain formation by means of directed capillary assembly. This way we achieved straight chains consisting of end-to-end aligned gold nanorods assembled in one specific direction with well-controlled gaps of ∼6 nm between the individual constituents. We determined the conditions for optimum quality and yield of nanorod chain assembly by investigating the influence of template dimensions and assembly temperature. In addition, we transferred the gold nanorod chains from the assembly template onto a Si/SiO2 target substrate, thus establishing a platform for a variety of nanoscale electronic and optical applications ranging from molecular electronics to optical and plasmonic devices. As a first example, electrical measurements are performed on contacted gold nanorod chains before and after their immersion in a solution of thiol end-capped oligophenylenevinylene molecules showing an increase in the conductance by three orders of magnitude, indicating molecular-mediated transport.


Enhanced phagocytosis and bactericidal activity of hepatic reticuloendothelial system during endotoxin tolerance.
The effects of tolerance to Escherichia coli endotoxin on the phagocytic and bactericidal activity of the hepatic reticuloendothelial system against viable E. coli were examined using ex vivo perfused rat livers. Livers were isolated from control and endotoxin-tolerant rats and perfused with a medium containing 5% homologous serum from either control or tolerant rats. After the addition of the E. coli (2 x 10(7) cells per ml) to the perfusate, the hepatic clearance of the bacteria was followed for 30 min. The highest activation of the hepatic reticuloendothelial system was observed when serum from tolerant animals was added to the perfusate. Under these conditions phagocytosis was 47% (12% in controls), and 37 to 38% of the bacteria were killed (5% in controls). This activation was less when livers obtained from tolerant rats were perfused with serum from controls or with saline only. The data suggests that, during endotoxin tolerance, humoral factors play an important role in the activation of the hepatic reticulendothelial system, although a direct stimulation of Kupffer cells also occurs. The enhancement of phagocytosis by tolerant serum did not require the presence of homologous antibodies and involved the activation of the alternative complement pathway, since it was lost after removal of factor B activity. On the other hand, stimulation of intracellular killing required both complement and specific antibodies. The data suggest a role of endotoxin in the activation of humoral and cellular mechanisms involved in the host resistance to gram-negative bacterial infection.


Early immune response against retrovirally transduced herpes simplex virus thymidine kinase-expressing gene-modified T cells coinfused with a T cell-depleted marrow graft: an altered immune response?
Administration of herpes simplex thymidine kinase (HSV-tk)-expressing, gene-modified T cells (GMCs) with T cell-depleted bone marrow transplantation (TCD-BMT) can allow modulation of posttransplantation alloreactivity. Twelve patients received 2 x 10(5) to 2 x 10(6) CD3+ donor GMCs per kilogram with HLA-identical sibling TCD-BMT. Despite extensive T cell depletion of bone marrow, an intensive conditioning regimen, and immunosuppressive graft-versus-host disease (GvHD) prophylaxis, infusion at the time of TCD-BMT of this low number of GMCs sufficed to induce a rapid GMC-specific immune response, as detected by interferon- enzyme- linked immunospot assay in six of eight patients, preferentially targeting HSV-tk. Maximal responses were reached early (median time, 49 [35-68] days post-BMT), with a subsequent rapid and significant decrease in five of six evaluable patients. Immune responses were negatively correlated with the maximal circulating GMC counts. However, such immune response did not result in the elimination of circulating GMCs and was not associated with measurable ex vivo cytotoxic activity against GMCs. Furthermore, alloreactive GMCs still could induce GCV-sensitive GvHD in one patient despite an ongoing immune response. Overall, infusion of HSV-tk-expressing GMCs at the time of BMT results in an early immune response. Such immune response may be altered and may not prevent persistent GCV-sensitive alloreactivity.


Factors influencing the latency of simple reaction time.
Simple reaction time (SRT), the minimal time needed to respond to a stimulus, is a basic measure of processing speed. SRTs were first measured by Francis Galton in the 19th century, who reported visual SRT latencies below 190 ms in young subjects. However, recent large-scale studies have reported substantially increased SRT latencies that differ markedly in different laboratories, in part due to timing delays introduced by the computer hardware and software used for SRT measurement. We developed a calibrated and temporally precise SRT test to analyze the factors that influence SRT latencies in a paradigm where visual stimuli were presented to the left or right hemifield at varying stimulus onset asynchronies (SOAs). Experiment 1 examined a community sample of 1469 subjects ranging in age from 18 to 65. Mean SRT latencies were short (231, 213 ms when corrected for hardware delays) and increased significantly with age (0.55 ms/year), but were unaffected by sex or education. As in previous studies, SRTs were prolonged at shorter SOAs and were slightly faster for stimuli presented in the visual field contralateral to the responding hand. Stimulus detection time (SDT) was estimated by subtracting movement initiation time, measured in a speeded finger tapping test, from SRTs. SDT latencies averaged 131 ms and were unaffected by age. Experiment 2 tested 189 subjects ranging in age from 18 to 82 years in a different laboratory using a larger range of SOAs. Both SRTs and SDTs were slightly prolonged (by 7 ms). SRT latencies increased with age while SDT latencies remained stable. Precise computer-based measurements of SRT latencies show that processing speed is as fast in contemporary populations as in the Victorian era, and that age-related increases in SRT latencies are due primarily to slowed motor output.


Lactate dehydrogenase values and bone scans as predictors of bone marrow involvement in small-cell lung cancer.
The value of serum lactate dehydrogenase (LDH) levels and bone scan results in predicting marrow involvement in small-cell carcinoma of the lung (SCCL) was studied. Records of 79 patients with SCCL who had undergone 92 bone marrow examinations were reviewed. None of 25 patients with marrow involvement had a normal LDH and a negative bone scan, compared with 23 of 59 in the uninvolved group. We conclude that patients with SCCL with both negative bone scans and normal LDH values have a less than 5% chance of marrow involvement.


Effect and placebo effect of acupressure (P6) on nausea and vomiting after outpatient gynaecological surgery.
Acupuncture and acupressure have previously been reported to possess antiemetic effect. We wanted to investigate the "true" and placebo effect of acupressure in prevention of postoperative nausea and vomiting (PONV). Sixty women undergoing outpatient minor gynaecological surgery were entered into a double-blind and randomised study. One group received acupressure with bilateral stimulation of P6 (A), a second group received bilateral placebo stimulation (P) and a third group received no acupressure wrist band and served as a reference group (R). PONV was evaluated as number of patients with complete response (no PONV), nausea only or vomiting. In addition, the need for rescue antiemetic medication and nausea after 24 h was registered. Complete response was obtained in 11, 11 and 9 patients in groups, A, P and R, respectively. Nine, 7 and 6 patients had nausea before discharge home, and 1, 1 and 8 patients were nauseated (8 vs 1 patient: P < 0.05) 24 h after operation in A, P and R groups, respectively. When compared to placebo acupressure (2 patients vomited and 5 needed rescue), significantly (P < 0.05) fewer needed rescue antiemetic medication after acupressure at P6 (no vomiting or rescue medication). When compared to the observation group (5 vomited and 4 needed rescue antiemetics), significantly fewer vomited after acupressure (P < 0.05) In patients undergoing brief gynaecological surgery, placebo effect of acupressure decreased nausea after 24 h but vomiting and need of rescue antiemetics was reduced only by acupressure with the correct P6 point stimulation.


Targeting the Lipid Metabolic Pathways for the Treatment of Malaria.
The control and eventual eradication of human malaria is considered one of the most important global public health goals of the 21st Century. Malaria, caused by intraerythrocytic protozoan parasites of the genus Plasmodium, is by far the most lethal and among the most prevalent of the infectious diseases. Four species of Plasmodium (P. falciparum, P. malariae, P. ovale, and P. vivax) are known to be infectious to humans, and more recent cases of infection due to P. knowlesi also have been reported. These species cause approximately 300 million annual cases of clinical malaria resulting in around one million deaths mostly caused by P. falciparum. The rapid emergence of drug-resistant Plasmodium strains has severely reduced the potency of medicines commonly used to treat and block the transmission of malaria and threatens the effectiveness of combination therapy in the field. New drugs that target important parasite functions, which are not the target of current antimalarial drugs, and have the potential to act against multi-drug-resistant Plasmodium strains are urgently needed. Recent studies in P. falciparum have unraveled new metabolic pathways for the synthesis of the parasite phospholipids and fatty acids. The present review summarizes our current understanding of these pathways in Plasmodium development and pathogenesis, and provides an update on the efforts underway to characterize their importance using genetic means and to develop antimalarial therapies targeting lipid metabolic pathways.


Risk Management in Executive Levels of Healthcare Organizations: Insights from a Scoping Review (2018).
This study attempted to present a framework and appropriate techniques for implementing risk management (RM) in executive levels of healthcare organizations (HCOs) and grasping new future research opportunities in this field. A scoping review was conducted of all English language studies, from January 2000 to October 2018 in the main bibliographic databases. Review selection and characterization were performed by two independent reviewers using pretested forms. Following a keyword search and an assessment of fit for this review, 37 studies were analyzed. Based on the findings and considering the ISO31000 model, a comprehensive yet simple framework of risk management is developed for the executive levels of HCOs. It includes five main phases: establishing the context, risk assessment, risk treatment, monitoring and review, and communication and consultation. A set of tools and techniques were also suggested for use at each phase. Also, the status of risk management in the executive levels of HCOs was determined based on the proposed framework. The framework can be used as a training tool to guide in effective risk assessment as well as a tool to assess non-clinical risks of healthcare organizations. Managers of healthcare organizations who seek to ensure high quality should use a range of risk management methods and tools in their organizations, based on their need, and not assume that each tool is comprehensive.


A review on creatinine measurement techniques.
This paper reviews the entire recent global tendency for creatinine measurement. Creatinine biosensors involve complex relationships between biology and micro-mechatronics to which the blood is subjected. Comparison between new and old methods shows that new techniques (e.g. Molecular Imprinted Polymers based algorithms) are better than old methods (e.g. Elisa) in terms of stability and linear range. All methods and their details for serum, plasma, urine and blood samples are surveyed. They are categorized into five main algorithms: optical, electrochemical, impedometrical, Ion Selective Field-Effect Transistor (ISFET) based technique and chromatography. Response time, detection limit, linear range and selectivity of reported sensors are discussed. Potentiometric measurement technique has the lowest response time of 4-10 s and the lowest detection limit of 0.28 nmol L(-1) belongs to chromatographic technique. Comparison between various techniques of measurements indicates that the best selectivity belongs to MIP based and chromatographic techniques.


Recycling of wastewater-derived phosphorus in Swedish agriculture--a proposal.
In 2001 the Swedish Government commissioned the Swedish Environmental Protection Agency (Swedish EPA) (i) to examine the need for stricter human health and environmental regulations governing the agricultural use of sewage sludge and (ii) to propose national targets for the agricultural recycling of phosphorus (P) originating from wastewater. The Swedish EPA may propose: (i) stricter regulations on sludge treatment to limit the risk of spreading pathogens in the environment and transmission of infectious diseases; (ii) recycling of 20-30% of wastewater-P to agriculture by 2015 and recycling of 35-50% of wastewater-P to agriculture by 2025.


Assessment of motion error for frame-based and noninvasive mask-based fixation using the Leksell Gamma Knife Icon radiosurgery system.
OBJECTIVEThe Leksell Gamma Knife Icon (GK Icon) radiosurgery system can utilize cone-beam computed tomography (CBCT) to evaluate motion error. This study compares the accuracy of frame-based and frameless mask-based fixation using the Icon system.METHODSA retrospective cohort study was conducted to evaluate patients who had undergone radiosurgery with the GK Icon system between June and December 2017. Patients were immobilized in either a stereotactic head frame or a noninvasive thermoplastic mask with stereotactic infrared (IR) camera monitoring. Setup error was defined as displacement of the skull in the stereotactic space upon setup as noted on pretreatment CBCT compared to its position in the stereotactic space defined by planning MRI for frame patients and defined as skull displacement on planning CBCT compared to its position on pretreatment CBCT for mask patients. For frame patients, the intrafractionation motion was measured by comparing pretreatment and posttreatment CBCT. For mask patients, the intrafractionation motion was evaluated by comparing pretreatment CBCT and additional CBCT obtained during the treatment. The translational and rotational errors were recorded.RESULTSData were collected from 77 patients undergoing SRS with the GK Icon. Sixty-four patients underwent frame fixation, with pre- and posttreatment CBCT studies obtained. Thirteen patients were treated using mask fixation to deliver a total of 33 treatment fractions. Mean setup and intrafraction translational and rotation errors were small for both fixation systems, within 1 mm and 1° in all axes. Yet mask fixation demonstrated significantly larger intrafraction errors than frame fixation. Also, there was greater variability in both setup and intrafraction errors for mask fixation than for frame fixation in all translational and rotational directions. Whether the GK treatment was for metastasis or nonmetastasis did not influence motion uncertainties between the two fixation types. Additionally, monitoring IR-based intrafraction motion for mask fixation-i.e., the number of treatment stoppages due to reaching the IR displacement threshold-correlated with increasing treatment time.CONCLUSIONSCompared to frame-based fixation, mask-based fixation demonstrated larger motion variations. The variability in motion error associated with mask fixation must be taken into account when planning for small lesions or lesions near critical structures.


Surgical Outcomes of Endoscopic Conjunctivodacryocystorhinostomy Using a Castroviejo Double-Ended Lacrimal Dilator.
The aim of this study was to describe the surgical method of endoscopic conjunctivodacryocystorhinostomy with Jones tube insertion using a Castroviejo double-ended lacrimal dilator and to elucidate the surgical outcomes. Under general anesthesia and preoperative epinephrine soaking, a monopolar needle cautery instrument was used to remove the nasal mucosa over the lacrimal and maxillary bone junction. After the lacrimal and maxillary bone junction was exposed, an oval osteotomy was formed. A Castroviejo double-ended lacrimal dilator was then inserted to create a direct fistula from the conjunctiva to the nasal cavity through the bony ostium. The dilator was grasped and withdrawn using smooth forceps to determine the tube length. The selected tube was then inserted into the fistula with a guide probe. Following removal of the probe, the inserted tube was fixed with 7-0 Ethilon suturing. Among 39 patients, a total of 49 cases were examined. The success rate was 73.4% (36/49 eyes). The average surgical time was 29.1 minutes for single-eye operations and 47.3 minutes for double-eye operations. Lateral migration (6/13; 46.2%), medial migration (3/13; 23.1%), granulation tissue obstruction (2/13; 15.4%), inflammation (1/13; 7.7%), and malpositioning (1/13; 7.7%) were the noted complications that led to reoperation. In conclusion, surgical management of endoscopic conjunctivodacryocystorhinostomy using a Castroviejo double-ended lacrimal dilator has several advantages. Using this device, easier surgical procedure, shorter surgical time, and more favorable success rate can be achieved without serious complications.


Apoptosis of primary-culture rat microglial cells induced by pathogenic Acanthamoeba spp.
To determine whether trophozoites and lysates of pathogenic Acanthamoeba spp. induce apoptosis in primary-culture microglial cells, transmission electron microscopic (TEM) examinations, assessment of DNA fragmentation by agarose gel electrophoresis, and the TdT-mediated dUTP nick-end labeling assay were performed. When a trophozoite of pathogenic Acanthamoeba culbertsoni came in contact with a microglial cell, the digipodium was observed by TEM. Nuclear chromatin condensation was observed in 10% of microglial cells, while it was not revealed when they were cocultured with weakly pathogenic Acanthamoeba royreba trophozoites. DNA fragmentation in microglial cells cocultured with the A. culbertsoni lysate was detected by electrophoresis, showing DNA ladder formation, whereas it was hardly observed in microglial cells cocultured with A. royreba. DNA fragmentation of microglial cells was also confirmed by flow cytometry analysis. The fluorescence of TdT-stained apoptotic bodies became intensely visible with microglial cells cocultured with the A. culbertsoni lysate. In contrast, with microglial cells cocultured with the A. royreba lysate, only a background level of fluorescence of TdT-stained apoptotic bodies was detected. These results suggest that some rat microglial cells cocultured with pathogenic A. culbertsoni undergo cytopathic changes which show the characteristics of the apoptotic process, such as nuclear condensation and DNA fragmentation.


Dural arteriovenous shunts involving the sphenoparietal sinus: a case report.
A 40-year-old male suffered transient global amnesia and was diagnosed as having a very rare location of dural arteriovenous shunt (dAVS) involving the right sphenoparietal sinus. The feeding pedicles of dAVS were not only arising from the right external carotid system, including the middle meningeal artery, but also the right internal carotid system including the ophthalmic artery. The dAVS drained into the right sphenoparietal sinus, right vein of Labbe, and right basal vein of Rosenthal through numerous dilated temporal cortical veins. The arteriovenous shunt itself was located at the dura around the right superior ophthalmic fissure and a large varix received direct shunting blood flow from the right sphenoparietal sinus. The dAVS was cured by drainer clipping following transarterial embolisations. A dAVS with cortical venous drainage is considered to have a higher risk of haemorrhage and venous infarction than the others and thus warrants treatment even if the clinical symptom is minimal. If the dAVS has a single drainer, it will be easily cured by way of drainer clipping. Preoperative embolisation makes clipping easier and safer. Intraoperative angiography is useful because preoperative angiography may not necessarily show the complete angioarchitecture around the lesion.


Serine/threonine acetylation of TGFβ-activated kinase (TAK1) by Yersinia pestis YopJ inhibits innate immune signaling.
The Gram-negative bacteria Yersinia pestis, causative agent of plague, is extremely virulent. One mechanism contributing to Y. pestis virulence is the presence of a type-three secretion system, which injects effector proteins, Yops, directly into immune cells of the infected host. One of these Yop proteins, YopJ, is proapoptotic and inhibits mammalian NF-κB and MAP-kinase signal transduction pathways. Although the molecular mechanism remained elusive for some time, recent work has shown that YopJ acts as a serine/threonine acetyl-transferase targeting MAP2 kinases. Using Drosophila as a model system, we find that YopJ inhibits one innate immune NF-κB signaling pathway (IMD) but not the other (Toll). In fact, we show YopJ mediated serine/threonine acetylation and inhibition of dTAK1, the critical MAP3 kinase in the IMD pathway. Acetylation of critical serine/threonine residues in the activation loop of Drosophila TAK1 blocks phosphorylation of the protein and subsequent kinase activation. In addition, studies in mammalian cells show similar modification and inhibition of hTAK1. These data present evidence that TAK1 is a target for YopJ-mediated inhibition.


Future precipitation increase from very high resolution ensemble downscaling of extreme atmospheric river storms in California.
Precipitation extremes will likely intensify under climate change. However, much uncertainty surrounds intensification of high-magnitude events that are often inadequately resolved by global climate models. In this analysis, we develop a framework involving targeted dynamical downscaling of historical and future extreme precipitation events produced by a large ensemble of a global climate model. This framework is applied to extreme "atmospheric river" storms in California. We find a substantial (10 to 40%) increase in total accumulated precipitation, with the largest relative increases in valleys and mountain lee-side areas. We also report even higher and more spatially uniform increases in hourly maximum precipitation intensity, which exceed Clausius-Clapeyron expectations. Up to 85% of this increase arises from thermodynamically driven increases in water vapor, with a smaller contribution by increased zonal wind strength. These findings imply substantial challenges for water and flood management in California, given future increases in intense atmospheric river-induced precipitation extremes.


Regulatory T-cell expansion and function do not account for the impaired alloreactivity of ex vivo-expanded T cells.
CD3- and CD28-activated T cells expanded for 12 days ex vivo to produce suicide gene-modified T cells are hyporesponsive to alloantigens. To investigate whether this impaired alloreactivity is a result of preferential expansion of regulatory T (Treg) cells, we compared peripheral blood mononuclear cells (PBMC) activated with CD3 and CD28 antibodies co-immobilized on beads and expanded for 12 days with interleukin (IL)-2 (Co(CD3/CD28) cells) to the respective unactivated PBMC in terms of proliferation, cytokine production, and expression of Treg markers [cytotoxic T-lymphocyte antigen 4 (CTLA4), glucocorticoid-induced tumour necrosis factor receptor (GITR) and forkhead box P3 (FoxP3)] after allostimulation. Alloreactive cells were identified by carboxyfluoresceine succinimidyl ester staining dilution. Alloreactive cells in Co(CD3/CD28) cells had a lower proliferative response and a lower potential for IL-2 and interferon-gamma secretion than did those in PBMC, demonstrating a functional impairment of alloreactive cells during ex vivo expansion. Expression of Treg markers transiently increased during ex vivo expansion and was unaffected by depletion of CD25(+) cells (containing Treg cells) before ex vivo PBMC expansion. Such prior CD25(+) depletion did not restore the alloreactivity of Co(CD3/CD28) cells. After allostimulation, expression of Treg markers was restricted to proliferative (alloreactive) cells among PBMC or Co(CD3/CD28) cells. Lastly, CD4(+) CD25(+) cells purified from Co(CD3/CD28) cells lacked suppressive activity when used as a third party, in contrast to CD4(+) CD25(+) cells purified from PBMC. In conclusion, the impaired alloreactivity of T cells expanded ex vivo is not a result of preferential Treg cell expansion and/or enhanced suppressive Treg activity.


Failure to elicit seroresponses to pneumococcal surface proteins (pneumococcal histidine triad D, pneumococcal choline-binding protein A, and serine proteinase precursor A) in children with pneumococcal bacteraemia.
Pneumococcal surface proteins (PSPs) elicit antibody responses in infants and young children exposed to Streptococcus pneumoniae. These seroresponses could contribute to the aetiological diagnosis of pneumococcal disease, e.g. during the clinical development of novel PSP-based vaccines. In this study, we assessed the kinetics of antibody responses to three highly conserved and immunogenic PSPs (pneumococcal histidine triad D (PhtD), pneumococcal choline-binding protein A (PcpA), and serine proteinase precursor A (PrtA)) in 106 children (median age, 21.3 months; males, 58.5%) admitted for pneumococcal bacteraemia. Anti-PhtD, anti-PcpA and anti-PrtA antibodies were measured by ELISA, and compared in 61 pairs of acute (≤7 days) and convalescent (>14 days of admission) serum samples. Acute serum titres were similar to those observed in healthy children, and were unaffected by the acid dissociation of circulating immune complexes. Despite proven bacteraemia, seroresponses (≥2-fold increase in anti-PSP antibody concentrations) were only identified in 31 of 61 children (50.8%), directed against PrtA (n = 23, 37.7%), PcpA (n = 19, 31.1%), and PhtD (n = 16, 26.2%), or several PSPs (two PSPs, n = 13, 21.3%; three PSPs, n = 7, 11.5%). Certain seroresponses were very strong (maximal fold-increases: PhtD, 26; PcpA, 72; PrtA, 12). However, anti-PSP antibody concentrations failed to increase in the convalescent sera of 30 of 61 (49.2%) bacteraemic children, and even declined (≥2 fold) in 13 of 61 (21.3%), mostly infants aged <6 months (8/13, 61.5%), possibly through consumption of maternal antibodies. Thus, pneumococcal bacteraemia may fail to elicit antibody responses, and may even have an antibody-depleting effect in infants. This novel observation identifies an important limitation of serology-based studies for the identification of bacteraemic children.


Domain-specific retrieval of source information in the medial temporal lobe.
Memory for context information (source memory) has been reported to rely on structures in the medial temporal lobe (MTL). Perirhinal cortex (anterior MTL) and parahippocampal cortex (posterior MTL) have distinct connectivity patterns with sensory neocortex, suggesting a possible modality-dependent organization of memory processes. The present study investigated the neural substrates of two different types of source information of newly encoded material using functional magnetic resonance imaging: auditory (speaker voice) and visual (texture and colour). Source judgements during retrieval were reliably above chance level for both modalities and performance did not differ between the auditory and visual condition. During encoding, activity predictive of subsequent source recollection was observed in the anterior hippocampus/parahippocampal gyrus, irrespective of source modality. During retrieval, on the other hand, a regional dissociation emerged: bilateral parahippocampal cortex discriminated between correct and incorrect auditory but not visual source judgements, whereas left perirhinal/entorhinal cortex showed the reverse pattern. These findings are consistent with recent lesion evidence of disrupted auditory but intact visual source memory following damage to the parahippocampal cortex. Results are discussed with respect to anatomical models of corticoparahippocampal connectivity and the functional organization of the MTL.


Familial hemiplegic migraine mutations change alpha1A Ca2+ channel kinetics.
Missense mutations in the pore-forming human alpha1A subunit of neuronal P/Q-type Ca2+ channels are associated with familial hemiplegic migraine (FHM). The pathophysiological consequences of these mutations are unknown. We have introduced the four single mutations reported for the human alpha1A subunit into the conserved rabbit alpha1A (R192Q, T666M, V714A, and I1819L) and investigated possible changes in channel function after functional expression of mutant subunits in Xenopus laevis oocytes. Changes in channel gating were observed for mutants T666M, V714A, and I1819L but not for R192Q. Ba2+ current (IBa) inactivation was slightly faster in mutants T666M and V714A than in wild type. The time course of recovery from channel inactivation was slower than in wild type in T666M and accelerated in V714A and I1819L. As a consequence, accumulation of channel inactivation during a train of 1-Hz pulses was more pronounced for mutant T666M and less pronounced for V714A and I1819A. Our data demonstrate that three of the four FHM mutations, located at the putative channel pore, alter inactivation gating and provide a pathophysiological basis for the postulated neuronal instability in patients with FHM.


Photochemical α-Aminonitrile Synthesis Using Zn-Phthalocyanines as Near-Infrared Photocatalysts.
While photochemical transformations with sunlight almost exclusively utilize the UV-vis part of the solar spectrum, the majority of the photons emitted by the sun have frequencies in the near-infrared region. Phthalocyanines show high structural similarity to the naturally occurring light-harvesting porphyrins, chlorins, and mainly bacteriochlorins and are also known for being efficient and affordable near-infrared light absorbers as well as triplet sensitizers for the production of singlet oxygen. Although having been neglected for a long time in synthetic organic chemistry due to their low solubility and high tendency toward aggregation, their unique photophysical properties and chemical robustness make phthalocyanines attractive photocatalysts for the application in near-infrared-light-driven synthesis strategies. Herein, we report a cheap, simple, and efficient photocatalytic protocol, which is easily scalable under continuous-flow conditions. Various phthalocyanines were studied as near-infrared photosensitizers in oxidative cyanations of tertiary amines to generate α-aminonitriles, a synthetically versatile compound class.


[The effect of chaotropic anions on the course of acid hemolysis in an isotonic sugar medium].
The influence of CNS-, ClO4-, NO3-, and SO4(2-) on the kinetics of acid hemolysis of erythrocytes of man, lamb, rat and rabbit was investigated. These chaotropic anions exercised strong and specific effects on the extent and time course of acid hemolysis under such conditions. In accordance with the intensity of the exercised antihemolytic effect, these anions formed a row coinciding with the Hoffmeister's row of their chaotropic action. The results demonstrate the impact of intermolecular interactions disturbance applied mainly on the cell membrane on the mechanism of acid hemolysis.


Homologue replacement in the import motor of the mitochondrial inner membrane of trypanosomes.
Many mitochondrial proteins contain N-terminal presequences that direct them to the organelle. The main driving force for their translocation across the inner membrane is provided by the presequence translocase-associated motor (PAM) which contains the J-protein Pam18. Here, we show that in the PAM of Trypanosoma brucei the function of Pam18 has been replaced by the non-orthologous euglenozoan-specific J-protein TbPam27. TbPam27 is specifically required for the import of mitochondrial presequence-containing but not for carrier proteins. Similar to yeast Pam18, TbPam27 requires an intact J-domain to function. Surprisingly, T. brucei still contains a bona fide Pam18 orthologue that, while essential for normal growth, is not involved in protein import. Thus, during evolution of kinetoplastids, Pam18 has been replaced by TbPam27. We propose that this replacement is linked to the transition from two ancestral and functionally distinct TIM complexes, found in most eukaryotes, to the single bifunctional TIM complex present in trypanosomes.


Healthy children ready to learn: an essential collaboration between health and education.
The "Healthy Children Ready to Learn" initiative starts with the underlying concept that health is a critical partner to optimum education. All children have a right to be healthy. At a minimum, this right assumes promoting optimum use of available and effective preventive measures, such as ensuring compliance with immunization recommendations; promoting measures to prevent injuries; ensuring opportunities to identify disease and disabilities early; and providing prompt treatment when needed. Families must receive the support and assistance they need to raise healthy and educated children. Activities directed toward National Education Goals and the related National Health Promotion and Disease Prevention Objectives can advance progress toward school readiness, focus attention and available resources on needed programs and services, and thus help the nation in achieving its goal of having all children arriving at school each day healthy, well nourished, and ready to learn. To realize these goals and objectives, the two critical systems of greatest importance to children, those providing health services and education, need to collaborate, not only among themselves, but also with social services. A range of critical health problems will require our attention if the goals are to be met, such as availability of prenatal care, infant mortality, inadequate nutrition during pregnancy or early childhood, or both, disease prevention by immunization, infants who have been exposed to drugs, fetal alcohol syndrome, and the emotional and mental disorders of early childhood, to name a few. At any one time, any family may be in need of appropriate services. To address the health and well-being of their young children, a continuum of appropriate, accessible services must be available in the community. The first steps toward successful achievement of the readiness goal will require the identification of health, education, and social service programs that serve young children and their families, and the creation of a climate that fosters innovative and effective collaboration between programs at the Federal and State levels, especially as it pertains to the community. Policies and programs should be built around the needs of families. In this regard, the critical role that parents play in shaping a healthy environment conducive to school readiness must be recognized as a key element in shaping the strategies that should help in achieving the readiness goal. Similarly important is the need to engage professional organizations and other private sector groups involved with health, education, and other children's issues to work with government and families to achieve the school readiness goal and its related health objectives.


Oleic acid decreases BCRP mediated efflux of mitoxantrone in Caco-2 cell monolayers.
Breast cancer resistance protein (BCRP) efflux restricts intestinal absorption of substances like heterocyclic amines, mycotoxins and certain human and veterinary drugs. Fat rich meals seem to increase absorption of drugs which are BCRP substrates or inhibitors. We therefore hypothesize that absorption of toxicants normally effluxed by BCRP are increased by fatty acids in food. Transport across and accumulation of 3H-Mitoxantrone (MXR) in Caco-2 cell monolayers were measured after 60 min exposure to emulsions of 3H-MXR (1μM) and oleic acid (0.5-5 mM). In addition, BCRP gene expression (RT-PCR) and the amount of BCRP protein (Western blot) were measured in oleic acid exposed Caco-2 cells. Oleic acid increased transport of MXR in a concentration dependent manner and 2 mM oleic acid or higher increased accumulation of MXR in cells, without any signs of cytotoxicity. Gene expression of BCRP was increased after exposure to oleic acid for 6 h, but the amount of BCRP protein was not increased. In conclusion, oleic acid clearly induced BCRP gene expression and reduced BCRP mediated efflux, although the amount of BCRP in cells was not affected. Consequently, effects of fatty acids on BCRP mediated efflux are important to consider in risk assessment of toxicants in food.


Novel stop mutation causing familial hypercholesterolemia in a Costa Rican family.
Combined heteroduplex single-strand conformation polymorphism (HEX-SSCP) analysis of the promoter and coding region of the low density lipoprotein receptor (LDLR) gene revealed a novel C to T mutation at nucleotide position 2056 in a Costa Rican patient with heterozygous familial hypercholesterolemia (FH). This nonsense mutation, Q665X, results in a termination codon in the epidermal growth factor (EGF) precursor homology domain of the mature LDLR.


Effects of 17 beta-estradiol exposure on Xenopus laevis gonadal histopathology.
The natural estrogen 17 beta-estradiol (E2) is a potential environmental contaminant commonly employed as a positive control substance in bioassays involving estrogenic effects. The aquatic anuran Xenopus laevis is a frequent subject of reproductive endocrine disruptor research; however, histopathological investigations have tended to be less than comprehensive. Consequently, a study was designed to characterize gross and microscopic changes in the gonads of X. laevis as a result of E2 exposure. Additional goals of this study, which consisted of three separate experiments, included the standardization of diagnostic terminology and criteria, the validation of statistical methodology, and the establishment of a half maximal effective concentration (EC50) for E2 as defined by an approximately 50% conversion of presumptive genotypic males to phenotypic females. In the first experiment, frogs were exposed to nominal concentrations of 0, 0.2, 1.5, or 6.0 microg/L E2. From these experimental results and those of a subsequent range finding trial, the EC50 for E2 was determined to be approximately 0.2 microg/L. This E2 concentration was utilized in the other two experiments, which were performed at different facilities to confirm the reproducibility of results. Experiments were conducted according to Good Laboratory Practice guidelines, and the histopathologic evaluations were peer reviewed by an independent pathologist. Among the three trials, the histopathological findings that were strongly associated with E2-exposure (p<0.001 to 0.0001) included an increase in the proportion of phenotypic females, mixed sex, dilated testis tubules, dividing gonocytes in the testis, and dilated ovarian cavities in phenotypic ovaries. A comparison of the gross and microscopic evaluations suggested that some morphologic changes in the gonads may potentially be missed if studies rely entirely on macroscopic assessment.


How reinforcement sensitivity and perceived risk influence young drivers' reported engagement in risky driving behaviors.
Gray's reinforcement sensitivity theory (RST), implementing Carver and White's behavior inhibition system (BIS) and behavior approach system (BAS) scales, was used to predict reported engagement in 10 risky driving behaviors: speeding (2 levels), driving under the influence of alcohol, racing other vehicles, cell phone use (hand-held and hands free), tailgating, unsafe overtaking, driving while fatigued, and not wearing a seatbelt. Participants were 165 young male and female (n=101) drivers aged 17-25 years who held a valid Australian driver's license. Effects of the explanatory variables and specific risk perceptions upon engagement in the reported risky driving behaviors were examined using SEM analyses. Also of interest was whether perceived risk mediated the relationship between the personality variables and reported engagement in risky driving behaviors. RST variables, negative reactivity, reward responsiveness and fun seeking, accounted for unique variance in young drivers' perceived risk. Reward responsiveness and perceived risk accounted for unique variance in young drivers' reported engagement in risky driving behaviors. Negative reactivity was completely mediated by perceived risk in its negative relationship with reported engagement. To better understand driving related risk decision making, future research could usefully incorporate drivers' motivation systems. This has the potential to lead to more tailored approaches to identifying risk-prone drivers and provide information for the development and implementation of media campaigns and educational programs.


A Wide Spread Red Lesion on the Surface of the Tongue-A Case Report and Review of Differential Diagnosis.
The tongue is considered to be the strongest muscular organ in the body and a very important structure in the oral cavity. It is involved in various critical functions such as taste, speech, chewing and deglutition. It has been known to be a harbinger of systemic health and disease. Lesions occurring on the tongue vary widely in colour and consistency. Widespread red lesions on the tongue need to be differentiated carefully from each other for a proper diagnosis and treatment and must be followed up for recurrence. The present case report demonstrates a widespread longstanding recurrent red lesion occurring on the surface of the tongue, its diagnosis and management.


Examining Choice and Control for People With IDD Over Time.
Choice making is an important aspect of everyone's life in terms of fully becoming an adult within a democratic society. People with intellectual and developmental disabilities (IDD) are at risk for diminished choice making due to various factors, including guardianships; dependence on supports that are not person-centered; and, in some cases, limited capacity to express one's desires effectively. Independent Monitoring for Quality (IM4Q) data for 9,195 and 9,817 for adult services users with IDD were analyzed across two types of choice. Repeated measures mixed regression examined choice over time after controlling for age, support needs, residence type, and community type. We found significant increases in everyday choice making among IDD service users in Pennsylvania, but not in support-related choice. This study is the first to our knowledge to consider change in choice making, an important indicator of rights and inclusion for persons with IDD. By comparing three waves of data from the state of Pennsylvania (2013, 2016, and 2019), we were able to detect changes in choice making over time among home and community-based service (HCBS) users with IDD.


Public Opinions and Concerns Regarding the Canadian Prime Minister's Daily COVID-19 Briefing: Longitudinal Study of YouTube Comments Using Machine Learning Techniques.
During the COVID-19 pandemic in Canada, Prime Minister Justin Trudeau provided updates on the novel coronavirus and the government's responses to the pandemic in his daily briefings from March 13 to May 22, 2020, delivered on the official Canadian Broadcasting Corporation (CBC) YouTube channel. The aim of this study was to examine comments on Canadian Prime Minister Trudeau's COVID-19 daily briefings by YouTube users and track these comments to extract the changing dynamics of the opinions and concerns of the public over time. We used machine learning techniques to longitudinally analyze a total of 46,732 English YouTube comments that were retrieved from 57 videos of Prime Minister Trudeau's COVID-19 daily briefings from March 13 to May 22, 2020. A natural language processing model, latent Dirichlet allocation, was used to choose salient topics among the sampled comments for each of the 57 videos. Thematic analysis was used to classify and summarize these salient topics into different prominent themes. We found 11 prominent themes, including strict border measures, public responses to Prime Minister Trudeau's policies, essential work and frontline workers, individuals' financial challenges, rental and mortgage subsidies, quarantine, government financial aid for enterprises and individuals, personal protective equipment, Canada and China's relationship, vaccines, and reopening. This study is the first to longitudinally investigate public discourse and concerns related to Prime Minister Trudeau's daily COVID-19 briefings in Canada. This study contributes to establishing a real-time feedback loop between the public and public health officials on social media. Hearing and reacting to real concerns from the public can enhance trust between the government and the public to prepare for future health emergencies.


Are You Saving for Retirement? Racial/Ethnic Differentials in Contributory Retirement Savings Plans.
How individuals and families accumulate retirement resources during working years is a key aspect of aging with implications for later life. This study examines how much, and by what mechanisms, savings in retirement plans vary by race/ethnicity. Using representative survey data and linked W-2 tax records, we estimate the probability of participation in employer-sponsored defined contribution (DC) retirement plans with probit regression, and contribution levels with ordinary least squares (OLS) models. We use Heckman models to adjust for potential sample selection. Black and Hispanic workers have lower participation and contributions in employer-sponsored DC retirement plans than do white workers, while Asian Americans have higher levels. The bulk of racial/ethnic differences is attributed to socioeconomic position, especially education and labor market circumstances like earnings. Differentials are also associated with family circumstances, namely for black workers. After accounting for education, labor market, and family covariates, social-psychological factors appear to explain only small portions of differences, especially for black and Hispanic. This study clarifies how racial/ethnic disparities in socioeconomic circumstances generate advantages and disadvantages in retirement wealth accumulation. Lower DC retirement plan participation and contributions among minorities in work life represent an underappreciated earlier-life channel through which racial inequalities in income and wealth in later life are generated.


[The pandemic--an afterthought].
During the second half of 2009 the Norwegian people experienced the first pandemic influenza since the Hong-Kong disease in 1968. The pandemic was initially believed to be severe, but it soon became evident that the disease was milder than anticipated. In spite of this the health authorities increased the level of threat and recommended general vaccination and exempt of oseltamivir (Tamiflu) from prescription requirements. In the paper we critically evaluate the evidence-base and consequences of these recommendations.


Combination of RNAseq and RADseq to Identify Physiological and Adaptive Responses to Acidification in the Eastern Oyster (Crassostrea virginica).
Ocean acidification (OA) is a major stressor threatening marine calcifiers, including the eastern oyster (Crassostrea virginica). In this paper, we provide insight into the molecular mechanisms associated with resilience to OA, with the dual intentions of probing both acclimation and adaptation potential in this species. C. virginica were spawned, and larvae were reared in control or acidified conditions immediately after fertilization. RNA samples were collected from larvae and juveniles, and DNA samples were collected from juveniles after undergoing OA-induced mortality and used to contrast gene expression (RNAseq) and SNP (ddRADseq) profiles from animals reared under both conditions. Results showed convergence of evidence from both approaches, particularly in genes involved in biomineralization that displayed significant changes in variant frequencies and gene expression levels among juveniles that survived acidification as compared to controls. Downregulated genes were related to immune processes, supporting previous studies demonstrating a reduction in immunity from exposure to OA. Acclimation to OA via regulation of gene expression might confer short-term resilience to immediate threats; however, the costs may not be sustainable, underscoring the importance of selection of resilient genotypes. Here, we identified SNPs associated with survival under OA conditions, suggesting that this commercially and ecologically important species might have the genetic variation needed for adaptation to future acidification. The identification of genetic features associated with OA resilience is a highly-needed step for the development of marker-assisted selection of oyster stocks for aquaculture and restoration activities.


Development of a low-cost airborne ultrasound sensor for the detection of brick joints behind a wall painting.
Non-destructive methods are of great interest for the analysis of cultural heritage. Among the different possible techniques, this paper presents a low cost prototype based on the emission and reception of airborne ultrasound without direct contact with the test specimen. We successfully performed a method test for the detection of brick joints under a XV th century Renaissance fresco of the Metropolitan Cathedral of the city of Valencia (Spain). Both laboratory and in situ results are in agreement. Using this prototype system, an early moisture detection system has been installed in the dome that supports the fresco. The result is encouraging and opens interesting prospects for future research.


Subepicardial haematoma, a rare and potentially lethal complication of CTO-PCI: case of an exceptional recovery after conservative management.
We present the case of an 82-year-old woman undergoing high-risk chronic total occlusion percutaneous coronary intervention (CTO-PCI) of the right coronary artery. Hours after the procedure, a subepicardial haematoma was diagnosed as a result of coronary perforation during the procedure. This rare and potentially lethal complication evolved exceptionally benignly after conservative management; our patient fully recovered. Increasingly complex procedures in high-risk patient categories warrant awareness of procedural complications, especially those that are subtle and appear relatively late, and are therefore most hazardous. Recognition of this rare complication and choosing the optimal strategy is of the utmost importance when dealing with patients who undergo PCI. We here describe the rare case of a potential lethal complication in high-risk CTO-PCI, which evolved relatively benignly.


A 5' RNA stem-loop participates in the transcription attenuation mechanism that controls expression of the Bacillus subtilis trpEDCFBA operon.
The trp RNA-binding attenuation protein (TRAP) regulates expression of the Bacillus subtilis trpEDCFBA operon by transcription attenuation. Tryptophan-activated TRAP binds to the nascent trp leader transcript by interacting with 11 (G/U)AG repeats. TRAP binding prevents formation of an antiterminator structure, thereby promoting formation of an overlapping terminator, and hence transcription is terminated before RNA polymerase can reach the trp structural genes. In addition to the antiterminator and terminator, a stem-loop structure is predicted to form at the 5' end of the trp leader transcript. Deletion of this structure resulted in a dramatic increase in expression of a trpE'-'lacZ translational fusion and a reduced ability to regulate expression in response to tryptophan. By introducing a series of point mutations in the 5' stem-loop, we found that both the sequence and the structure of the hairpin are important for its regulatory function and that compensatory changes that restored base pairing partially restored wild-type-like expression levels. Our results indicate that the 5' stem-loop functions primarily through the TRAP-dependent regulatory pathway. Gel shift results demonstrate that the 5' stem-loop increases the affinity of TRAP for trp leader RNA four- to fivefold, suggesting that the 5' structure interacts with TRAP. In vitro transcription results indicate that this 5' structure functions in the attenuation mechanism, since deletion of the stem-loop caused an increase in transcription readthrough. An oligonucleotide complementary to a segment of the 5' stem-loop was used to demonstrate that formation of the 5' structure is required for proper attenuation control of this operon.


Electrical Activity of Powerhouse Muscles During the Teaser Exercise of Pilates Using Different Types of Apparatus.
We compared the electrical activity of certain powerhouse muscles-External Oblique, Multifidus, Adductor Longus, and Gluteus Medius-during the teaser exercise of the Pilates Method, performed on various types of apparatus-the Mat, Reformer, and Wall Unit. Fifteen female practitioners of the Classic Pilates Method (32.6 ± 7.7 years old; 21.9 ± 1.9 body mass index) performed the teaser in each situation while electromyographic (EMG) and kinematic data were collected. Root mean square values of the flexion phase were compared. All muscles showed higher EMG activity in Reformer compared with Wall Unit, and Multifidus, Adductor Longus, and Gluteus Medius showed higher EMG activity in Mat compared with Wall Unit. No difference was found between Reformer and Mat.


Heteropoda toxin 2 interaction with Kv4.3 and Kv4.1 reveals differences in gating modification.
Kv4 (Shal) potassium channels are responsible for the transient outward K(+) currents in mammalian hearts and central nervous systems. Heteropoda toxin 2 (HpTx2) is an inhibitor cysteine knot peptide toxin specific for Kv4 channels that inhibits gating of Kv4.3 in the voltage-dependent manner typical for this type of toxin. HpTx2 interacts with four independent binding sites containing two conserved hydrophobic amino acids in the S3b transmembrane segments of Kv4.3 and the closely related Kv4.1. Despite these similarities, HpTx2 interaction with Kv4.1 is considerably less voltage-dependent, has smaller shifts in the voltage-dependences of conductance and steady-state inactivation, and a 3-fold higher K(d) value. Swapping four nonconserved amino acids in S3b between the two channels exchanges the phenotypic response to HpTx2. To understand these differences in gating modification, we constructed Markov models of Kv4.3 and Kv4.1 activation gating in the presence of HpTx2. Both models feature a series of voltage-dependent steps leading to a final voltage-independent transition to the open state and closely replicate the experimental data. Interaction with HpTx2 increases the energy barrier for channel opening by slowing activation and accelerating deactivation. The greater degree of voltage-dependence in Kv4.3 occurs because it is the voltage-dependent transitions that are most affected by HpTx2; in contrast, it is the voltage-independent step in Kv4.1 that is most affected by the presence of toxin. These data demonstrate the basis for subtype-specificity of HpTx2 and point the way to a general model of gating modifier toxin interaction with voltage-gated ion channels.


Magnetic resonance angiographic evaluation of circle of Willis: A morphologic study in a tertiary hospital set up.
Anatomy of circle of Willis (CW) shows wide variation in different individuals, population groups, and has vital clinical significance in causation and presentation of clinical disease. This study evaluates the anatomical variations, incidence of various common anomalies of CW in south Indian tertiary hospital set up, using three-dimensional time-of-flight (3D-TOF) magnetic resonance angiography (MRA). A total of 300 patients referred for neuroimaging study over a period of 2-year were included in the analysis. In this prospective and retrospective study, 198 men and 102 women; mean age, 55 years) underwent 3D-TOF MR angiograms of the CW using a 1.5-tesla MR scanner. Images were reviewed for anatomical configuration of the CW using maximum intensity projection (MIP) and 3D volume rendered images. On analysis, a complete CW was seen in 50 (16.6%) of 300 subjects. An incomplete anterior and posterior CW was found in 66 (22%) The remaining 184 (61.3%) subjects had partially complete CW configuration. The most common type of CW in a single subject was anterior variant type A and posterior type variant E. We observed wide variation in CW configuration in our patients. The prevalence of complete configuration of the circle is 16.6%; slightly higher in females and younger subjects. Complete anterior circle was present in 77.3%. Most common anterior variant is type A (normal anterior configuration) with a prevalence of 66%. The most common posterior circle variant is type E (hypoplasia or absence of both PcomA) with 32.6%. Overall, CW variants are slightly more common among the women in comparison to men. Incidence of associated anomalies like aneurysm or arteriovenous malformation (AVM) was comparable to that described in literature.


Establishment and application of high throughput screening model for hepatitis C virus NS3-4A protease inhibitors in vitro.
To establish fluorescence resonance energy transfer (FRET) assay method of detecting proteolytic activity of non-structural protein 3-4A (NS3-4A) serine protease of hepatitis C virus (HCV) for high throughput screening inhibitors against HCV in vitro. HCV recombinant plasmid pMAL~c2/NS3-4A was transformed into the E.coli strain K12TB1. Maltose-binding-protein (MBP) NS3-4A fusion protein expression was induced by adding isopropyl-β-D-thiogalacto-pyranoside (IPTG) and purified by affinity chromatography. The proteolytic activity of MBP-NS3-4A protease was analyzed by FRET with the special protease substrate. The reaction system in this model was optimized, and the reliability of the model was evaluated. High throughput screening model for HCV NS3-4A protease inhibitors was established, and the best concentrations of enzyme and substrate were optimized. In the model, the Km value of protease was 4.74 μmol/L, Z factor was up to 0.80, and coefficient of variation (CV) was 1.91%. BILN 2061, one of the known HCV protease inhibitors, was measured with the Ki of 0.30 nmol/L. The assay model using FRET method for HCV NS3 4A serine protease is stable and reliable, and the model is suitable for high throughput screening for HCV NS3 4A protease inhibitors.


Bacterial Lipopolysaccharide Destabilizes Influenza Viruses.
Depending on the specific viral pathogen, commensal bacteria can promote or reduce the severity of viral infection and disease progression in their hosts. Influenza A virus (IAV) has a broad host range, comprises many subtypes, and utilizes different routes of transmission, including the fecal-oral route in wild birds. It has been previously demonstrated that commensal bacteria can interact with the host's immune system to protect against IAV pathogenesis. However, it is unclear whether bacteria and their products may be interacting directly with IAV to impact virion stability. Herein we show that gastrointestinal (GI) tract bacterial isolates in an in vitro system significantly reduce the stability of IAV. Moreover, bacterial lipopolysaccharide (LPS), found on the exterior surfaces of bacteria, was sufficient to significantly decrease the stability of both human and avian viral strains in a temperature-dependent manner, including at the relevant temperatures of their respective hosts and the external aquatic habitat. The subtype and host origin of the viruses were shown to affect the extent to which IAV was susceptible to LPS. Furthermore, using a receptor binding assay and transmission electron microscopy, we observed that LPS binds to and alters the morphology of influenza virions, suggesting that direct interaction with the viral surface contributes to the observed antiviral effect of LPS on influenza. IMPORTANCE Influenza A virus (IAV), transmitted primarily via the fecal-oral route in wild birds, encounters high concentrations of bacteria and their products. Understanding the extent to which bacteria affect the infectivity of IAV will lead to a broader understanding of viral ecology in reservoir hosts and may lead to insights for the development of therapeutics in respiratory infection. Herein we show that bacteria and lipopolysaccharide (LPS) interact with and destabilize influenza virions. Moreover, we show that LPS reduces the long-term persistence and freeze-thaw stability of IAV, which is important information for modeling the movement and emergence of novel strains from animal hosts. Our results, demonstrating that the subtype and host origin of a virus also influence its susceptibility to LPS, raise key questions about the fitness of viruses in reservoir hosts, their potential to transmit to humans, and the importance of bacterial-viral interactions in viral ecology.


Trypanosoma cruzi prevalence and clinical forms in blood donor candidates in Brazil.
The prevalence and clinical forms of Trypanosoma cruzi were evaluated among blood donor candidates attended at a general hospital in Rio de Janeiro, Brazil, from January 1997 to April 1999. The investigation was done by means of the indirect hemagglutination test and was confirmed via ELISA. Data were collected from clinical examinations, conventional electrocardiogram, chest radiography and echocardiography. The results showed that despite Trypanosoma cruzi prevalence of 1.17% (128 patients), mainly in males aged 40 years or over, 70.8% of these patients, mainly males aged 19 to 39 years, demonstrated abnormalities that allowed the diagnosis of cardiopathy and/or esophagopathy. This once again corroborates the importance of Trypanosoma cruzi infection in urban centers.


Platelet dropping, bleeding and new treatment requirements in ITP patients after inactivated COVID-19 vaccination.
Significant decreases in platelet counts and ITP relapses have been documented in ITP patients receiving COVID-19 mRNA vaccines; however, the effect of the inactivated COVID-19 vaccine on ITP patients remains unclear. The present study aimed to investigate the impact of inactivated COVID-19 vaccines on ITP patients, with a focus on platelet dropping events, bleeding events/scores, and the requirement of a new round of treatment. A total of 118 ITP patients, with 97 chronic ITP and 21 persistent ITP, who received inactivated COVID-19 immunization were investigated retrospectively. Following vaccination (within 1 month), ITP patients reported platelet dropping (31.36 %), new bleeding events (22.88 %), increases in bleeding scores (23.73 %), and new treatment requirements (22.03 %). Among them, persistent ITP patients with disease duration of 3-12 months had higher ratios of the above adverse events (71.43 %, 57.14 %, 61.90 % and 71.43 %, respectively) than chronic ITP patients with duration > 1 year (22.68 %, 15.46 %, 15.46 % and 11.34 %, respectively); patients' disease duration was negatively correlated with platelet dropping events and new treatment requirements. Furthermore, logistic regression analysis also supported the above findings, revealing that persistent ITP patients had 9.40-9.70, 7.24-10.08, and 27.17-28.51 times incidence of having platelet dropping events, new bleeding events, and new treatment requirements after vaccination, respectively, when compared to chronic ITP patients. In conclusion, the present study demonstrates that after receiving inactivated COVID-19 vaccines, ITP patients may experience platelet dropping, which may lead to new bleeding events and the requirement of a new round of treatment for ITP recurrence. As a result, platelet level monitoring is crucial for ITP patients during the vaccination, especially those with persistent ITP.


Geometric constructions for predicting hough transform performance.
Graphical techniques are presented for estimating Hough transform performance for detecting straight lines in noisy digital pictures. The methods show explicitly the influence of noise due, for example, to quantizing errors on the accuracy of estimating the underlying sets of collinear points. Exact bounds are obtained for sets of collinear points showing that under quite general conditions only the endpoints are of significance. Theorems governing grid size for the accumulator method are given.


A Novel Form Factor For PPG-based Blood Pressure Monitoring.
Blood pressure (BP) is one of the four main vital signs in medicine and may be a useful signal for wellness tracking and for user-aware interfaces in human-computer interaction. The current standard for BP measurement uses cuff-based devices that block an artery temporarily to get a single, discrete measurement of BP. Recently, there have been significant efforts to measure correlates of BP continuously and non-invasively from relevant signals like photoplethysmography (PPG), which responds to volumetric changes in arteries due to blood pulsations. In this paper, we explore a novel setup with two points of instrumentation, one on the head and a second on the wrist, for recording PPG and estimating the pulse wave velocity, which is a major correlate of BP, along with other waveform-related features. We prospectively tested the device on 10 subjects who followed a protocol for the deliberate variation of BP while ground truth measurements were taken using a reference cuff-device. Generic absolute BP models, which use the collected data for leave-one-subject-out cross-validation, yielded an error of -0.14 ± 7.3 mmHg for systolic BP (SBP) and -0.21±6.7 mmHg for diastolic BP (DBP), which are within the regulatory limits of 5 ± 8 mmHg. Notably, the correlation between the predicted BPs and the ground truth BPs was higher for SBP (r = 0.74, p < 0.001) than for DBP (r = 0.34, p < 0.001). The results show that the proposed form factor can extract BP-related features that could be used for continuous, cuff-less BP monitoring.


Modulation of human airway smooth muscle proliferation by type 3 phosphodiesterase inhibition.
Elevation in cell cAMP content can inhibit mitogenic signaling in cultured human airway smooth muscle (HASM) cells. We studied the effects of the type 3-selective phosphodiesterase inhibitor siguazodan, the type 4-selective phosphodiesterase inhibitor rolipram, and the nonselective inhibitor 3-isobutyl-1-methylxanthine (IBMX) on proliferation of cultured HASM cells. At concentrations selective for the type 3 phosphodiesterase isoform, siguazodan inhibited both [3H]thymidine incorporation (IC50 2 microM) and the increase in cell number (10 microM; 64% reduction) induced by platelet-derived growth factor-BB (20 ng/ml). These effects were mimicked by IBMX. At concentrations selective for type 4 phosphodiesterase inhibition, rolipram was without effect. A 20-min exposure to siguazodan and rolipram did not increase whole cell cAMP levels. However, in HASM cells transfected with a cAMP-responsive luciferase reporter (p6CRE/Luc), increases in cAMP-driven luciferase expression were seen with siguazodan (3.9-fold) and IBMX (16.5-fold). These data suggest that inhibition of the type 3 phosphodiesterase isoform present in airway smooth muscle results in inhibition of mitogenic signaling, possibly through an increase in cAMP-driven gene expression.


Global travel and Gram-negative bacterial resistance; implications on clinical management.
Antimicrobial resistance (AR) is escalating worldwide with the potential for dire consequences, global travel contributes to the dissemination of resistant pathogens from one region to another. The World Health Organization identified the rapid emergence and prevalence of carbapenem-resistant Gram-negative species, including Enterobacterales, Acinetobacter baumannii, and Pseudomonas aeruginosa, as an international crisis due to treatment challenges, poor health outcomes, increased mortality, and high economic costs caused by these pathogens. This review describes key carbapenem-resistant (CR) Gram-negative species, changes in current global and regional trends, AR surveillance and reporting, and identifies drivers of change, specifically travel. Finally, we review clinical implications and challenges of treating CR infections which exist due to widespread dissemination of CR bacteria. A literature search was conducted using PubMed, Google Scholar, Ebsco, and ProQuest (from 2000 to December 2019). The level of global travel is increasing, and antimicrobial resistance continues to disseminate worldwide. Healthcare providers risk assessment for AR needs to consider a patient's recent travel history, including pre-travel and intra-travel antimicrobial prescription, and potential exposure based on geography. Patient education, healthcare provider awareness, and access to data and surveillance resources are critical to inform antimicrobial selection and improve health outcomes.


Receptor-mediated increases in cytosolic Ca2+ in the human erythroleukaemia cell line involve pertussis toxin-sensitive and -insensitive pathways.
The pluripotent human erythroleukaemia cell line, HEL, possesses erythrocytic, megakaryocytic and macrophage-like properties. With respect to signal transduction, HEL cells have been used as a model system for platelets, but little attention has been paid to their phagocytic properties. We studied the effects of various receptor agonists on the intracellular free Ca2+ concentration ([Ca2+]i) in HEL cells. Thrombin, platelet-activating factor (PAF), ATP, UTP, prostaglandins E1 and E2 (PGE1 and PGE2), the PGE2 analogue sulprostone and the stable PGI2 analogues iloprost and cicaprost increased [Ca2+]i. ADP was less effective than ATP, and UDP was unable to increase [Ca2+]i. The increases in [Ca2+]i induced by thrombin, PAF, ATP, UTP, iloprost and cicaprost were pertussis toxin-insensitive, whereas the increases induced by PGE2 and sulprostone were completely inhibited by the toxin. The increase in [Ca2+]i induced by PGE1 was partially inhibited by pertussis toxin. PGE2 did not desensitize the increase in [Ca2+]i induced by iloprost, and vice versa. PGE1 desensitized the response to PGE2 and iloprost but not vice versa. Adrenaline potentiated the iloprost- but not the PGE2-induced rise in [Ca2+]i. The phorbol ester phorbol 12-myristate 13-acetate completely blocked the rise in [Ca2+]i induced by ATP and PGE1, whereas the increases induced by thrombin and PAF were only partially inhibited. Agonists increased [Ca2+]i through release from internal stores and sustained Ca2+ influx. Thrombin stimulated Mn2+ influx, which was blocked by Ni2+. Diltiazem, isradipine, gramicidin and 1-(beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl)-1H-imidazole hydrochloride (SK&F 96365) did not affect agonist-induced rises in [Ca2+]i. HEL cells contained substantial amounts of beta-glucuronidase which, however, could not be released, and they did not aggregate or generate superoxide. Our data suggest that: (1) HEL cells possess nucleotide receptors with properties similar to those of phagocytes; (2) they possess receptors for PGE2 and PGI2, and PGE1 is an agonist at both receptors; (3) agonist-induced increases in [Ca2+]i are mediated through pertussis toxin-sensitive as well as -insensitive signal transduction pathways; and (4) agonists increase [Ca2+]i by mobilization from internal stores and influx from the extracellular space through cation channels with properties similar to those of phagocytes and platelets.


Menière's disease.
Menière's disease causes paroxysmal rotatory vertigo, due to endolymphatic hydrops, an accumulation of endolymph in the endolymphatic space of the labyrinth. Its major symptoms are attacks of rotatory vertigo lasting minutes to hours, with unilateral hearing loss, tinnitus and aural fullness. As the disease progresses, attacks happen less often, but hearing loss and tinnitus gradually become permanent. Neuro-otological complications may develop, such as benign paroxysmal positional vertigo, vestibular drop attacks and bilateral vestibulopathy. The diagnosis of Menière's disease is based on the typical clinical picture and typical findings on the audiogram. Furthermore, it is now possible to diagnose it by MR of the inner ear. Long-term management has several steps, including diet, diuretics, intratympanic injection of corticosteroid or gentamicin and surgery (endolymphatic sac surgery, grommet insertion, surgical labyrinthectomy).


The use of the neuronavigator in the orbital surgery of a rare case of Rosai-Dorfman disease.
Neuronavigators are robotic devices that can help to bridge the gap between the data of three-dimensional images and the object by means of interactive computerized programs. A rare case of Rosai-Dorfman disease with prevalent bilateral endo-orbital interest is reported, in which an assisting neuronavigation system was used. The navigation system used is a stereotaxic system without the use of supporting mechanical arms, permitting bi-dimensional and tridimensional reconstruction through the data that are obtained from diagnostic equipment such as computed tomography, magnetic resonance imaging, single photon emission computed tomography, and positron emission tomography. This system has given an accurate localization of the pathological findings and has permitted a precise evaluation of the relation between the lesion and intraconical structures, which are fundamental requisites to optimize the treatment and reduce the postsurgical complications. The exciting results achieved in cerebral parenchyma surgery motivate extension of the use of neuronavigator systems to the splanchnocranial complex.


Effects of the Collaborative Assessment and Management of Suicidality (CAMS) Model: A Secondary Analysis of Moderation and Influencing Factors.
In a randomized controlled trial, we found that suicidal patients who received Collaborative Assessment and Management of Suicidality (CAMS) reported greater improvements in suicidal ideation and mental health distress compared to participants who received treatment as usual (TAU). Here, we explored moderators and mediators of the effectiveness of CAMS. Compared to TAU, CAMS was more effective in reducing suicidal ideation when the working alliance, in particular its bond subcomponent, was low. In terms of reducing mental health distress, CAMS was superior to TAU only for participants who did not use illicit drugs and, more tentatively, only for patients without borderline personality traits. We suggest that CAMS may repair a difficult vantage point in terms of poor working alliance in patients with suicide ideation. To obtain superior benefits of CAMS upon more general mental health distress in patients with drug abuse or borderline traits, these problems may need to be more explicitly targeted in parallel.


Microvessel density and p53 in detecting cervical cancer by FDG PET in cases of suspected recurrence.
Cervical cancer is the second most frequently diagnosed cancer in women worldwide. About one-third of patients experience recurrent disease. A better chance of survival might be achieved by the early detection of recurrent cervical cancer. [(18)F]fluoro-2-deoxy-D-glucose (FDG) PET could be a promising imaging modality for this purpose, given that FDG PET has high diagnostic efficacy. Ideally, pre-selection of patients should be performed before considering FDG PET. The purpose of this study was to investigate parameters of primary cervical cancer associated with recurrence as a basis for pre-selection of patients in whom FDG PET should be performed. Thirty-eight cervical cancer patients, clinically suspected of having recurrent disease, underwent FDG PET. Tissue from primary tumours and nine histologically confirmed metastases was analysed for biomarkers possibly related to glucose metabolism and prognosis (vascular endothelial growth factor, CD31 for microvessel density, glucose transporter-1, hexokinases I, II and III, Ki67, p53, hypoxia-inducible factor 1alpha, and degree of infiltration by lymphocytes and macrophages). Based on clinical outcome, sensitivity and specificity of FDG PET were 96% and 100%, respectively. Cox regression revealed microvessel density and p53 (tumour suppressor protein) to be the two most important biomarkers for prediction of recurrence (hazard ratios 2.54 and 2.28, respectively). By combining these two biomarkers in a parallel test, sensitivity and specificity in predicting recurrence were 87% and 71%, respectively. Leave-one-out cross-validation demonstrated predictive validity of a model based on microvessel density and p53. In this first study of its kind, we have demonstrated that microvessel density and p53 profiles could be important in pre-selecting cervical cancer patients for detection of recurrence by FDG PET.


Ethical issues in m-Health applications in community health work in India - a scoping review.
Mobile phone-based interventions are being increasingly used in community health work in India. The extensive use of mobile phones in community health work is associated with several ethical issues. This review was conducted to identify the ethical issues related to mHealth applications in community health work in India. We performed a scoping review of literature in PubMed and Google Scholar using a search strategy that we developed. We included studies that mentioned ethical issues in mHealth applications that involved community health work and community health workers in India, published in peer reviewed English language journals between 2011 and 2021. All three authors screened the articles, shortlisted them, read them, and extracted the data. We then synthesised the data into a conceptual framework. Our search yielded 1125 papers, from which we screened and shortlisted 121, after reading which we included 58 in the final scoping review. The main ethical issues identified from review of these papers included benefits of mHealth applications such as improved quality of care, increased awareness about health and illness, increased accountability of the health system, accurate data capture and timely data driven decision making. The risks of mHealth applications identified were impersonal communication of community health worker, increased workload, potential breach in privacy, confidentiality, and stigmatisation. The inherent inequities in access to mobile phones in the community due to gender and class led to exclusion of women and the poor from the benefits of mHealth interventions. Though mHealth interventions increased access to healthcare by taking healthcare to remote areas through tele-health, unless we contextualise mHealth to local rural settings through community engagement, it is likely to remain inequitable. This scoping review revealed that there is a lack of well conducted empirical studies which explore the ethical issues related to mHealth applications in community health work.


Subclinical hypothyroidism is independently associated with microalbuminuria in a cohort of prediabetic egyptian adults.
Recent evidence has suggested an association between subclinical hypothyroidism (SCH) and microalbuminuria in patients with type 2 diabetes. However, whether SCH is related to microalbuminuria among subjects with prediabetes has not been studied. Thus, we evaluated the association between SCH and microalbuminuria in a cohort of prediabetic Egyptian adults. A total of 147 prediabetic subjects and 150 healthy controls matched for age and sex were enrolled in this study. Anthropometric measurements, plasma glucose, lipid profile, homeostasis model assessment of insulin resistance (HOMA-IR), thyroid stimulating hormone (TSH), free thyroxine, triiodothyronine levels, and urinary albumin-creatinine ratio (UACR) were assessed. The prevalence of SCH and microalbuminuria in the prediabetic subjects was higher than that in the healthy controls (16.3% vs. 4%, P<0.001; and 12.9% vs. 5.3%, P=0.02, respectively). Prediabetic subjects with SCH were characterized by significantly higher HOMA-IR, TSH levels, UACR, and prevalence of microalbuminuria than those with euthyroidism. TSH level was associated with total cholesterol (P=0.05), fasting insulin (P=0.01), HOMA-IR (P=0.01), and UACR (P=0.005). UACR was associated with waist circumference (P=0.01), fasting insulin (P=0.05), and HOMA-IR (P=0.02). With multiple logistic regression analysis, SCH was associated with microalbuminuria independent of confounding variables (β=2.59; P=0.01). Our findings suggest that prediabetic subjects with SCH demonstrate higher prevalence of microalbuminuria than their non-SCH counterparts. SCH is also independently associated with microalbuminuria in prediabetic subjects. Screening and treatment for SCH may be warranted in those patients.


High-performance liquid chromatography with column switching for the determination of cocaine and benzoylecgonine concentrations in vitreous humor.
A sample concentration technique was adapted for the determination of cocaine and benzoylecgonine (BE) concentrations in vitreous humor. Vitreous humor (0.5 mL) was diluted 1:1 with water and applied through a filter onto a 3-cm preconcentration column. Following a simple wash step, the analytes were flushed directly onto a reversed-phase analytical high-performance liquid chromatography (HPLC) system. Absolute recoveries were high (above 90%) and the chromatograms were free from interference. Analysis for the drug and its breakdown product was performed using ultraviolet (UV) visible photodiode array detection, which allowed confirmation of peak identity. Recognizable UV spectra could be measured with as little as 20 ng on column. Comparison of the drug levels in 27 blood and vitreous humor samples showed that, while there was only a low correlation between the blood and vitreous concentrations (R = 0.70), vitreous cocaine and BE determinations were good indicators of antemortem cocaine use. In almost all cases, the vitreous BE concentrations were higher than the cocaine concentrations. The technique was easy to perform and the vitreous samples were especially compatible with this low-labor analytical procedure.


The Prevention Effect of Selenium on Prevalence of Children Kaschin-Beck Disease in Active Endemic Areas in Qinghai Plateau.
Selenium deficiency is an important environmental risk factor of Kaschin-Beck disease (KBD), and appropriate selenium supplement can reduce the prevalence of KBD. Guide and Xinghai counties, active endemic areas of KBD in Qinghai Plateau, are characteristic with low level of selenium. The aim of this article was to explore the relationship between selenium content and prevalence of children KBD in some active endemic areas from Guide and Xinghai counties. The historical data of KBD were collected, including the detectable rates of KBD and selenium contents of the hair of children, and then the relationship between the prevalence of KBD and selenium contents of hair was analyzed. In KBD endemic areas of Guide County, the detectable rates of X-ray and metaphysic lesion were declined from 25.00 and 16.96% in 2000 to 13.75 and 13.75% in 2010, respectively. Similarly, in KBD endemic areas of Xinghai County, the detectable rates of X-ray and metaphysic lesion were declined from 46.51 and 40.31% in 2000 to 10.64 and 8.51% in 2010, respectively. The selenium contents of hair in Xinghai county were increased from 130.01 ± 48.08 μg/kg in 2003 to 211.8 ± 86.64 μg/kg in 2010(t = 2.98, P < 0.05); the selenium content of hair in Guide County were increased from 142.30 ± 62.02 μg/kg in 2003 to 182.09 ± 78.46 μg/kg in 2010 (t = 3.12, P < 0.05). There was a negative correlation between the prevalence of KBD and selenium contents of hair (r = -0.785). There was a close relationship between selenium content and prevalence of KBD. Selenium could reduce the prevalence of KBD, so it is very necessary to supplement selenium appropriately for KBD prevention.


Perturbing cohesin dynamics drives MRE11 nuclease-dependent replication fork slowing.
Pds5 is required for sister chromatid cohesion, and somewhat paradoxically, to remove cohesin from chromosomes. We found that Pds5 plays a critical role during DNA replication that is distinct from its previously known functions. Loss of Pds5 hinders replication fork progression in unperturbed human and mouse cells. Inhibition of MRE11 nuclease activity restores fork progression, suggesting that Pds5 protects forks from MRE11-activity. Loss of Pds5 also leads to double-strand breaks, which are again reduced by MRE11 inhibition. The replication function of Pds5 is independent of its previously reported interaction with BRCA2. Unlike Pds5, BRCA2 protects forks from nucleolytic degradation only in the presence of genotoxic stress. Moreover, our iPOND analysis shows that the loading of Pds5 and other cohesion factors on replication forks is not affected by the BRCA2 status. Pds5 role in DNA replication is shared by the other cohesin-removal factor Wapl, but not by the cohesin complex component Rad21. Interestingly, depletion of Rad21 in a Pds5-deficient background rescues the phenotype observed upon Pds5 depletion alone. These findings support a model where loss of either component of the cohesin releasin complex perturbs cohesin dynamics on replication forks, hindering fork progression and promoting MRE11-dependent fork slowing.


[Coffee and cardiovascular disease risk: yin and yang].
Many epidemiological studies have addressed the effects of coffee on cardiovascular disease. Most case-control studies suggest an increased risk in high coffee consumers, whereas cohort studies indicate no clear association with cardiovascular risk. Several aspects could be considered to explain and/or reconcile these inconsistencies. Selection bias and recall bias may explain a positive association supported by case-control studies. An inadequate adjustment for many confounding factors (i.e., smoking, poor diet, sedentary lifestyle, etc.) could also affect the relationship between coffee consumption and cardiovascular risk. Moreover, coffee contains several biologically active substances that may have either beneficial or harmful effects on the cardiovascular system. The development of complete/partial tolerance to some caffeine effects in habitual drinkers adds to the complexity of coffee effects. Variation in cup size and methods of coffee preparation may also explain some conflicting results. As it is not reasonable to conduct randomized controlled trials, it is recommended that coffee consumption be moderate in healthy people and limited in individuals at high risk.


Problems with provision: barriers to drinking water quality and public health in rural Tasmania, Australia.
Access to safe drinking water is essential to human life and wellbeing, and is a key public health issue. However, many communities in rural and regional parts of Australia are unable to access drinking water that meets national standards for protecting human health. The aim of this research was to identify the key issues in and barriers to the provision and management of safe drinking water in rural Tasmania, Australia. Semi-structured interviews were conducted with key local government employees and public health officials responsible for management of drinking water in rural Tasmania. Participants were asked about their core public health duties, regulatory responsibilities, perceptions and management of risk, as well as the key barriers that may be affecting the provision of safe drinking water. This research highlights the effect of rural locality on management and safety of fresh water in protecting public health. The key issues contributing to problems with drinking water provision and quality identified by participants included: poor and inadequate water supply infrastructure; lack of resources and staffing; inadequate catchment monitoring; and the effect of competing land uses, such as forestry, on water supply quality. This research raises issues of inequity in the provision of safe drinking water in rural communities. It highlights not only the increasing need for greater funding by state and commonwealth government for basic services such as drinking water, but also the importance of an holistic and integrated approach to managing drinking water resources in rural Tasmania.


Comparison of the pathogenesis of murine cytomegalovirus in lung and liver following intraperitoneal or intratracheal infection.
This study compares the pathogenesis of murine cytomegalovirus (MCMV) infections following intraperitoneal (i.p.) and intratracheal (i.t.) inoculation. No deaths were seen in mice given 10(6) p.f.u. MCMV i.t., whereas 52% mortality occurred among animals given this dose i.p. This difference in mortality was not due to different effects on the lung, since virus titres in this organ on progressive days post-infection were similar for the two routes of inoculation and similar, minor histopathological changes were observed. In contrast, virus titres in the livers of mice inoculated i.p. were 100-fold higher than for those inoculated i.t., and histopathological changes were noticeably greater in the i.p. group. This suggests that the mortality seen following i.p. inoculation may have been due, at least in part, to effects of viral infection on liver function. Parallels between various forms of human cytomegalovirus infections and the types of infections seen following i.t. and i.p. infection with MCMV were observed.


Investigating the Migratory Behavior of Soybean Looper, a Major Pest of Soybean, through Comparisons with the Corn Pest Fall Armyworm Using Mitochondrial Haplotypes and a Sex-Linked Marker.
The Noctuid moth soybean looper (SBL), Chrysodeixis includens (Walker), is an economically important pest of soybean (Glycine max (Linnaeus) Merrill). Because it is not known to survive freezing winters, permanent populations in the United States are believed to be limited to the southern regions of Texas and Florida, yet its geographical range of infestations annually extend to Canada. This indicates annual migrations of thousands of kilometers during the spring and summer growing season. This behavior is like that of the fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), also a Noctuid that is a major global pest of corn. SBL and FAW are projected to have very similar distributions of permanent populations in North America based on climate suitability modeling and the overlap in the distribution of their preferred host plants (corn and soybean). It therefore seems likely that the two species will display similar migratory behavior in the United States. This was tested by identifying genetic markers in SBL analogous to those successfully used to delineate FAW migratory pathways and comparing the distribution patterns of the markers from the two species. Contrary to expectations, the results indicate substantial differences in migratory behavior that appear to be related to differences in the timing of corn and soybean plantings. These findings underscore the importance of agricultural practices in influencing pest migration patterns, in particular the timing of host availability relative to mean seasonal air transport patterns.


Fever of unknown origin.
Fever of unknown origin (FUO) often is defined as a fever greater than 38.3 degrees C on several occasions during at least 3 weeks with uncertain diagnosis after a number of obligatory tests. In general, infection accounts for approximately one-fourth of cases of FUO, followed by neoplasm and noninfectious inflammatory diseases. No diagnosis is reached in up to 50% of cases. Scintigraphic methods, such as (67)Ga-citrate, labeled leukocytes, and (18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET), are often used in the diagnosis of FUO. In FUO, FDG-PET appears to be of great advantage because malignancy, inflammation, and infection can be detected. FDG-PET does not seem to contribute to a final diagnosis in patients with normal erythrocyte sedimentation rate and C-reactive protein. Image fusion combining PET and computed tomography facilitates anatomical localization of increased FDG uptake and better guiding for further diagnostic tests to achieve a final diagnosis. In conclusion, the body of evidence on the utility of FDG-PET in patients with FUO is growing and FDG-PET will probably become the preferred diagnostic procedure, especially when a definite diagnosis cannot easily be achieved. Because of favorable characteristics of FDG-PET, conventional scintigraphic techniques may be replaced by FDG-PET in institutions in which PET is available.


Training curriculum to help patient representatives participate meaningfully in the development of clinical practice guidelines.
Patient participation in the development of clinical practice guidelines (CPGs) is critical for validity and trust. Many guideline panels now include patient representatives. Engagement of these individuals may be improved by training them about the process and their role before they join a guideline panel. To aid patient representatives in engagement in the improvement of guidelines, we developed and implemented a curriculum. The curriculum was developed based on content from the Grading of Recommendations Assessment, Development and Evaluation (GRADE) Working Group and readability principles, and was delivered through a webinar followed by a face-to-face half a day workshop. Twenty-four patient representatives were recruited by the American Society of Hematology to serve on guideline panels. Barriers assessment was facilitated by a pre-curriculum survey. The curriculum targeted patient representatives' knowledge, skills and attitudes and was followed by actual engagement in a guideline panel and a post-curriculum survey. Participants reported that the combination of the two training methods was very useful (9/10 on the Likert scale) in increasing their knowledge about guideline development. They agreed that their skills and self-efficacy in developing guidelines improved (8/10). Their attitudes (confidence in their ability to participate) improved by 30% between the webinar and the workshop. They developed a script to use during panel deliberations and an instruction sheet for the guideline panel about how to empower and engage them as active participants in the guideline development process. The benefits of incorporating patients' voice in CPGs are multifold. These benefits may be optimised by providing patient representatives with training that addresses barriers to engagement and tools to increase their knowledge, skills and attitudes required for meaningful participation.


AIDS-related lymphoma in Brazil. Histopathology, immunophenotype, and association with Epstein-Barr virus.
The occurrence of malignant lymphoma is an increasingly important cause of morbidity and mortality in AIDS patients. The incidence of AIDS-related lymphoma in some developing countries such as Brazil is increasing as the survival of HIV infection has improved. Although there is a clear association between several types of immunodeficiency-related lymphomas and Epstein-Barr virus (EBV), the association of EBV infection in AIDS-related lymphoma in Brazil, where the incidence of AIDS is high, is unknown. Formalin-fixed, paraffin-embedded tissue from 24 cases of AIDS-related lymphoma in Brazil were analyzed for morphologic classification, immunophenotype, and EBV association using in situ hybridization studies with an EBV-EBER1 biotinylated probe. Twenty cases of AIDS-related lymphoma were classified as non-Hodgkin's lymphoma and four cases were Hodgkin's disease. Eleven non-Hodgkin's lymphomas were classified as diffuse large cell type, five cases were small non-cleaved cell, Burkitt-type, and four cases were large cell immunoblastic non-Hodgkin's lymphoma. Eighteen cases were of B-cell phenotype; one was a T-cell lymphoma, and one was classified as null. Epstein-Barr virus (EBV) was demonstrated in the majority of tumor cells of 11 of 20 (55%) of the cases non-Hodgkin's lymphomas and in 3 of 4 (75%) cases of Hodgkin's disease. AIDS-related lymphomas in Brazil are usually of large cell/immunoblastic type, but Hodgkin's disease is also seen. Both non-Hodgkin's lymphoma and Hodgkin's disease are often associated with EBV infection. The non-Hodgkin's lymphoma is predominantly of B-cell phenotype.