Functional expression of tagged human Na+-glucose cotransporter in Xenopus laevis oocytes.
1. High-affinity, secondary active transport of glucose in the intestine and kidney is mediated by an integral membrane protein named SGLT1 (sodium glucose cotransporter). Though basic properties of the transporter are now defined, many questions regarding the structure- function relationship of the protein, its biosynthesis and targeting remain unanswered. In order to better address these questions, we produced a functional hSGLT1 protein (from human) containing a reporter tag. 2. Six constructs, made from three tags (myc, haemaglutinin and poly-His) inserted at both the C- and N-terminal positions, were thus tested using the Xenopus oocyte expression system via electrophysiology and immunohistochemistry. Of these, only the hSGLT1 construct with the myc tag inserted at the N-terminal position proved to be of interest, all other constructs showing no or little transport activity. A systematic comparison of transport properties was therefore performed between the myc-tagged and the untagged hSGLT1 proteins. 3. On the basis of both steady-state (affinities for substrate (glucose) and inhibitor (phlorizin) as well as expression levels) and presteady-state parameters (transient currents) we conclude that the two proteins are functionally indistinguishable, at least under these criteria. Immunological detection confirmed the appropriate targeting of the tagged protein to the plasma membrane of the oocyte with the epitope located at the extracellular side. 4. The myc-tagged hSGLT1 was also successfully expressed in polarized MDCK cells. alpha-Methylglucose uptake studies on transfected cells showed an exclusively apical uptake pathway, thus indicating that the expressed protein was correctly targeted to the apical domain of the cell. 5. These comparative studies demonstrate that the myc epitope inserted at the N-terminus of hSGLT1 produces a fully functional protein while other epitopes of similar size inserted at either end of the protein inactivated the final protein.


One-stop hysteroscopy clinic for postmenopausal bleeding.
To evaluate the role and feasibility of the "1-stop" clinic for management of postmenopausal bleeding, based on the use of transvaginal ultrasound and hysteroscopic examination under local anesthesia. A retrospective analysis of 308 patients referred to the clinic between October 2000 and May 2002 was carried out. Endometrial thickness of > 4 mm, as measured by transvaginal scan, was considered the cutoff limit for further investigation by hysteroscopy. Three hundred eight patients were seen in the 1-stop clinic. One hundred ten (35.7%) had endometrial thickness < or = 4 mm with no other associated abnormal findings and were discharged without proceeding to hysteroscopy. In 8 patients the scan was not conclusive, and they were offered hysteroscopy. Another 4 had a fluid-filled uterine cavity on sonography and were offered dilatation and curettage under general anesthesia. Eight patients opted to have the hysteroscopy done under general anesthesia, and in 12, hysteroscopyfailed under local anesthesia. Outpatient hysteroscopy was performed in 174 cases; 86 (49.4%) had normalfindings and were discharged. Endometrial polyps were found in 55 (31.6%) patients; in 42 the polyp was successfully removed at the same setting. Twenty patients (11.5%) had submucous fibroids and were discharged. A suspicious lesion was found in 13 (7.5%) patients; histologic examination confirmed endometrial carcinoma. A total of 216 of 308 (70%) patients were totally managed and discharged at the first visit with no further follow-up, and hospital admission was avoided in 258 (83.7%) of cases. The 1-stop clinic is effective in reducing the number of hospital visits per patient as well as hospital admissions and the waiting list.


Investigation of geriatric syndromes associated with medication in Japan using insurance claims data.
Multiple risk factors are involved in geriatric syndrome (GS) occurring in older adults. Although drug therapy often contributes to GS, the specific causes among older adults in Japan remain unclear. In this study, we examined the possible prescribing cascade rate among older outpatients eligible for Late-stage Elderly Health Insurance and elucidated the differences between GS and GS associated with medication (GSAM) trends. This retrospective study enrolled patients from health insurance claims data in Japan between October 2018 and March 2019; hospitalized patients were excluded. Two groups were identified among the participants with GS: GS (no use of GS-causing medications) and possible-GSAM (p-GSAM; use of GS-causing medications). The collected data were analyzed using the Bell Curve for Excel, and statistical significance was set at P < 0.05. In total, 137 781 outpatients were enrolled. Of the 32 259 outpatients who did not use GS-causing medications, 7342 were classified into the GS group. Among 105 522 outpatients who used GS-causing medications, 8347 were classified as having p-GSAM. The mean number of prescriptions was significantly higher in the p-GSAM group than in the GS group (P < 0.01). Furthermore, all GS symptoms showed significant differences, with impaired appetite being the most prevalent in the p-GSAM group than in the GS group (P < 0.01). A possible prescribing cascade was suspected in 2826 (33.9%) of 8347 outpatients in the p-GSAM group. Impaired appetite in patients taking GS-causing medications might lead to prescribing cascades. Further studies are needed to prevent such prescribing cascades. Geriatr Gerontol Int 2023; ••: ••-••.


High Disparity Between Orthopedic Resident Interest and Participation in International Health Electives.
Few orthopedic surgical residency programs offer international health electives (IHEs). Efforts to expand these programs have been increasing across medical disciplines. Whether orthopedic residents will participate remains unknown. This study quantified and characterized orthopedic resident interest and barriers to IHEs in US residency programs. A web-based survey was administered to residents from 154 US orthopedic residency programs accredited by the Accreditation Council for Graduate Medical Education 2014 to 2015. Questions assessed demographics and program background, previous medical experience abroad, barriers to participation, and level of interest in participating in an international health elective during their training and beyond. Twenty-seven (17.5%) residency programs responded. Chi-square analysis showed that residents who expressed interest in participating were significantly more likely to have experience abroad compared with those who expressed no interest (P<.004). Analysis using Mann-Whitney U test suggested that those who expressed interest were more likely to believe IHEs are important to resident training (P<.0011; mean Likert scale score of 3.7 vs 2.6), provide valuable experience (P<.001; mean Likert scale score of 4.2 vs 3.2), and should be required for orthopedic residencies (P<.001; mean Likert scale score of 2.8 vs 1.9). Residents are strongly interested in participating in IHEs during their training, and many may integrate global health into future practices. Residents perceive lack of funding and scheduling flexibility as barriers preventing them from participating. Prior experience abroad influences level of interest, and international clinical experience may enhance future perception of its value. [Orthopedics. 2016; 39(4):e680-e686.].


Identification of a 52-kD calmodulin-binding protein associated with the mitotic spindle apparatus in mammalian cells.
A pool of 10 calmodulin-binding proteins (CBPs) was isolated from Chinese hamster ovary (CHO) cells via calmodulin (CaM)-Sepharose affinity chromatography. One of these ten isolated CBPs with a molecular mass of 52 kD was also found to be present in isolated CHO cell mitotic spindles. Affinity-purified antibodies generated against this pool of isolated CBPs recognize a single 52-kD protein in isolated CHO cell mitotic spindles by immunoblot analysis. Immunofluorescence examination of CHO, 3T3, NRK, PTK-2, and HeLa cells resulted in a distinct pattern of mitotic spindle fluorescence. The localization pattern of this 52-kD CBP directly parallels that of CaM in the spindle apparatus throughout the various stages of mitosis. Interestingly, there was no association of this 52-kD CBP with cytoplasmic microtubules. As is the case with CaM, the localization pattern of the 52-kD CBP in interphase cells is diffuse within the cytoplasm and is not associated with any discrete, cellular structures. This 52-kD CBP appears to represent the first mitotic spindle-specific calmodulin-binding protein identified and represents an initial step toward the ultimate determination of CaM function in the mitotic spindle apparatus.


Tissue- and paralogue-specific functions of acyl-CoA-binding proteins in lipid metabolism in Caenorhabditis elegans.
ACBP (acyl-CoA-binding protein) is a small primarily cytosolic protein that binds acyl-CoA esters with high specificity and affinity. ACBP has been identified in all eukaryotic species, indicating that it performs a basal cellular function. However, differential tissue expression and the existence of several ACBP paralogues in many eukaryotic species indicate that these proteins serve distinct functions. The nematode Caenorhabditis elegans expresses seven ACBPs: four basal forms and three ACBP domain proteins. We find that each of these paralogues is capable of complementing the growth of ACBP-deficient yeast cells, and that they exhibit distinct temporal and tissue expression patterns in C. elegans. We have obtained loss-of-function mutants for six of these forms. All single mutants display relatively subtle phenotypes; however, we find that functional loss of ACBP-1 leads to reduced triacylglycerol (triglyceride) levels and aberrant lipid droplet morphology and number in the intestine. We also show that worms lacking ACBP-2 show a severe decrease in the β-oxidation of unsaturated fatty acids. A quadruple mutant, lacking all basal ACBPs, is slightly developmentally delayed, displays abnormal intestinal lipid storage, and increased β-oxidation. Collectively, the present results suggest that each of the ACBP paralogues serves a distinct function in C. elegans.


Conservation phylogeography: does historical diversity contribute to regional vulnerability in European tree frogs (Hyla arborea)?
Documenting and preserving the genetic diversity of populations, which conditions their long-term survival, have become a major issue in conservation biology. The loss of diversity often documented in declining populations is usually assumed to result from human disturbances; however, historical biogeographic events, otherwise known to strongly impact diversity, are rarely considered in this context. We apply a multilocus phylogeographic study to investigate the late-Quaternary history of a tree frog (Hyla arborea) with declining populations in the northern and western part of its distribution range. Mitochondrial and nuclear polymorphisms reveal high genetic diversity in the Balkan Peninsula, with a spatial structure moulded by the last glaciations. While two of the main refugial lineages remained limited to the Balkans (Adriatic coast, southern Balkans), a third one expanded to recolonize Northern and Western Europe, loosing much of its diversity in the process. Our findings show that mobile and a priori homogeneous taxa may also display substructure within glacial refugia ('refugia within refugia') and emphasize the importance of the Balkans as a major European biodiversity centre. Moreover, the distribution of diversity roughly coincides with regional conservation situations, consistent with the idea that historically impoverished genetic diversity may interact with anthropogenic disturbances, and increase the vulnerability of populations. Phylogeographic models seem important to fully appreciate the risks of local declines and inform conservation strategies.


Pharmacologic and sensorimotor components of satiation in cigarette smoking.
To examine mechanisms underlying satiation in cigarette smoking, 18 smokers received intravenous (i.v.) nicotine, alone or in combination with denicotinized cigarette smoke. Nicotine was administered using programmed presentations of either pulsed injections or continuous infusions, with i.v. saline serving as a control. A high-nicotine cigarette smoke condition (usual brand) was also presented. During each of the six test sessions, subjects were allowed to puff on their usual brands of cigarette ad libitum while the programmed satiation conditions were in force. Administration of i.v. nicotine caused a small suppression of ad libitum smoking behavior; denicotinized smoke produced a significantly larger reduction, showing that short-term satiation is more dependent on the presentation of smoke than delivery of nicotine per se. However, denicotinized smoke alone did not have as much effect as puffs from the usual brands of cigarettes. The combination of i.v. nicotine and denicotinized smoke puffs produced equivalent satiation to that of the usual brand. Cigarette craving and negative affect were partially relieved by iv nicotine presentations as well as by denicotinized smoke, and again the combination of i.v. nicotine and denicotinized smoke approximated the effects of the usual brand. The results of this study underscore the importance of both sensorimotor aspects of smoking and the pharmacologic effects of nicotine in tobacco dependence.


The National Health Service and medical libraries in England.
This paper provides an overview of the National Health Service in England, including its history, its present structure, and factors that make it different from the U.S. health care system. The NHS libraries and librarians are discussed. The NHS is divided into fourteen regions in England. Three specific NHS libraries in the Wessex region are described--a hospital library, an administrative library, and a patient education library. The problems and accomplishments of NHS librarians are discussed, as well as some similarities between NHS libraries and U.S. medical libraries. Medical librarians are urged to try to understand the medical library scene in England, so that they can Learn from the unique strengths of their counterparts there.


Attention deficit hyperactivity disorder and developmental coordination disorder: Two separate disorders or do they share a common etiology.
Attention deficit hyperactivity disorder (ADHD) has been described as the most prevalent behavioral disorder in children. Developmental coordination disorder (DCD) is one of the most prevalent childhood movement disorders. The overlap between the two conditions is estimated to be around 50%, with both substantially interfering with functioning and development, and leading to poorer psychosocial outcomes. This review provides an overview of the relationship between ADHD and DCD, discussing the common presenting features, etiology, neural basis, as well as associated deficits in motor functioning, attention and executive functioning. It is currently unclear which specific motor and cognitive difficulties are intrinsic to each disorder as many studies of ADHD have not been screened for DCD and vice-versa. The evidence supporting common brain underpinnings is still very limited, but studies using well defined samples have pointed to non-shared underpinnings for ADHD and DCD. The current paper suggests that ADHD and DCD are separate disorders that may require different treatment approaches.


Biomechanical considerations for cycling interventions in rehabilitation.
Individuals with physical disabilities may benefit from cycling interventions, which could address impairments while potentially minimizing stress on joints. Improvements in impairments may then have an impact on mobility, activity, and participation. Cycling studies with adults and children who are healthy have shown that many factors can influence the biomechanics of cycling. These factors include seat height, crank arm length, foot position, cadence, and workload. Knowledge of these factors is important for rehabilitation professionals who prescribe cycling as an intervention for individuals with disabilities, because changing these factors can potentially influence the therapeutic outcomes. In addition, further research is needed to fully understand the effect of these factors on individuals with disabilities.


[Effect of ascorbic acid on the development of a spontaneous lymphoproliferative process in SJL/J-strain mice].
Ascorbic acid (AA) effects on the development of lymphoproliferative process according to the type of a reticular variant of Hodgkin's disease have been investigated in SJL/J mice. An oral AA injection in a 0.05% aqueous solution for 6-8 months (0.625 mg/kg) with a total dose of 0.5 per mouse has not produced any side effects, has reduced both the rate of tumour process development (from 80.5% in control to 68.0% in experiment) and the degree of its extension, and also has led to a statistically significant decrease in the frequency of development of general amyloidosis.


Endrin resistance in the pine mouse.
Wild pine mice with a history of treatment with endrin exhibited a 12-fold greater tolerance to the pesticide than did mice having no history of endrin treatment. A genetic basis for the resistance would be expected, in view of the fact that the resistant mice had been exposed to endrin for 11 years. However, sublethal dosages conferred a degree of resistance on both mouse populations; this indicates the possibility of an inducible resistance.


Visuo-perceptual organization and working memory in patients with schizophrenia.
We explore the mechanisms sub-tending the re-organization and memorization of visual information by studying how these mechanisms fail in patients with schizophrenia. Several studies have suggested that patients have difficulties in organizing information in perception and memory. We explore to what extent prompting patients to group items influences memory performance. We distinguish automatic grouping from top-down grouping processes, which are especially involved in re-organizing information. The main task was to memorize pairs of figures. Following manipulation of proximity, pairs of figures were part of the same perceptual group (within-group pair, formed on the basis of automatic grouping) or belonged to different groups (between-group pairs, re-grouped through top-down processes). Prior to the memory task, subjects ran a perception task prompting them to prioritize either within-group or between-group pairs. Unlike patients, controls globally benefited from grouping by proximity in the memory task. In addition, the results showed that prioritizing between-group pairs had a deleterious effect in patients, but with a large decrement in memory performance in the case of within-group rather than between-group figures. This occurred despite preserved focalization on within-group figures, as shown by eye-movement recordings. The suggestion is that when patients are prompted to re-group separate items, they can do so, but the benefit derived from automatic grouping is then not only lost but also reversed. This suggests re-organizing visual information not only involves re-grouping separate items but also integrating these new groups in a unified representation, which is impaired in patients with schizophrenia.


Expanded polytetrafluoroethylene as tendon replacement: an experimental study in chickens.
We designed an experimental animal model (in chickens) to assess the potential applications, above all in hand surgery, of an expanded polytetrafluoroethylene (e-PTFE) graft as a replacement for a natural tendon. The results of macroscopic, histological, and functional observations made five weeks, and three, four, five, and six months after implantation showed that the e-PTFE seemed to be a good replacement for tendons, because it integrated well with surrounding tissue and permitted good functional recovery within a reasonable time period.


Breaking Reciprocity with Space-Time-Coding Digital Metasurfaces.
Metasurfaces are artificially engineered ultrathin structures that can finely tailor and control electromagnetic wavefronts. There is currently a strong interest in exploring their capability to lift some fundamental limitations dictated by Lorentz reciprocity, which have strong implications in communication, heat management, and energy harvesting. Time-varying approaches have emerged as attractive alternatives to conventional schemes relying on magnetic or nonlinear materials, but experimental evidence is currently limited to devices such as circulators and antennas. Here, the recently proposed concept of space-time-coding digital metasurfaces is leveraged to break reciprocity. Moreover, it is shown that such nonreciprocal effects can be controlled dynamically. This approach relies on inducing suitable spatiotemporal phase gradients in a programmable way via digital modulation of the metasurface-elements' phase repsonse, which enable anomalous reflections accompanied by frequency conversions. A prototype operating at microwave frequencies is designed and fabricated for proof-of-concept validation. Measured results are in good agreement with theory, hence providing the first experimental evidence of nonreciprocal reflection effects enabled by space-time-modulated digital metasurfaces. The proposed concept and platform set the stage for "on-demand" realization of nonreciprocal effects, in programmable or reconfigurable fashions, which may find several promising applications, including frequency conversion, Doppler frequency illusion, optical isolation, and unidirectional transmission.


PET Imaging of Microglial Activation-Beyond Targeting TSPO.
Neuroinflammation, which involves microglial activation, is thought to play a key role in the development and progression of neurodegenerative diseases and other brain pathologies. Positron emission tomography is an ideal imaging technique for studying biochemical processes in vivo, and particularly for studying the living brain. Neuroinflammation has been traditionally studied using radiotracers targeting the translocator protein 18 kDa, but this comes with certain limitations. The current review describes alternative biological targets that have gained interest for the imaging of microglial activation over recent years, such as the cannabinoid receptor type 2, cyclooxygenase-2, the P2X₇ receptor and reactive oxygen species, and some promising radiotracers for these targets. Although many advances have been made in the field of neuroinflammation imaging, current radiotracers all target the pro-inflammatory (M1) phenotype of activated microglia, since the number of known biological targets specific for the anti-inflammatory (M2) phenotype that are also suited as a target for radiotracer development is still limited. Next to proceeding the currently available tracers for M1 microglia into the clinic, the development of a suitable radiotracer for M2 microglia would mean a great advance in the field, as this would allow for imaging of the dynamics of microglial activation in different diseases.


Sustained thromboprophylaxis mediated by an RBC-targeted pro-urokinase zymogen activated at the site of clot formation.
Plasminogen activators (PAs) are used to treat life-threatening thrombosis, but not for thromboprophylaxis because of rapid clearance, risk of bleeding, and central nervous system (CNS) toxicity. We describe a novel strategy that may help to overcome these limitations by targeting a thrombin-activated PA pro-drug to circulating red blood cells (RBCs). We fused a single chain antibody (scFv Ter-119) that binds to mouse glycophorin A (GPA) with a variant human single-chain low molecular weight urokinase construct that can be activated selectively by thrombin (scFv/uPA-T). scFv/uPA-T bound specifically to mouse RBCs without altering their biocompatibility and retained its zymogenic properties until converted by thrombin into an active 2-chain molecule. As a result, RBC-bound scFv/uPA-T caused thrombin-induced fibrinolysis. One hour and 48 hours after intravenous (IV) injection in mice, approximately 70% and approximately 35% of scFv/uPA-T was retained in the blood, respectively, and approximately 95% of the circulating scFv/uPA-T remained bound to RBCs. A single IV injection of scFv/uPA-T provided effective prophylaxis against arterial and venous thrombosis for up to 24 hours. Thus, prophylactic delivery of RBC-targeted PA pro-drugs activated selectively at the site of clot formation represents a new approach to prevent thrombosis in clinical settings where the risk of clotting is high.


Solid lipid nanoparticles as oral delivery systems of phenolic compounds: Overcoming pharmacokinetic limitations for nutraceutical applications.
Drug delivery systems, accompanied by nanoparticle technology, have recently emerged as prominent solutions to improve the pharmacokinetic properties, namely bioavailability, of therapeutic and nutraceutical agents. Solid lipid nanoparticles (SLNs) have received much attention from researchers due to their potential to protect or improve drug properties. SLNs have been reported to be an alternative system to traditional carriers, such as emulsions, liposomes, and polymeric nanoparticles. Phenolic compounds are widespread in plant-derived foodstuffs and therefore abundant in our diet. Over the last decades, phenolic compounds have received considerable attention due to several health promoting properties, mostly related to their antioxidant activity, which can have important implications for health. However, most of these compounds have been associated with poor bioavailability being poorly absorbed, rapidly metabolized and eliminated, which compromises its biological and pharmacological benefits. This paper provides a systematic review of the use of SLNs as oral delivery systems of phenolic compounds, in order to overcome pharmacokinetic limitations of these compounds and improved nutraceutical potential. In vitro studies, as well as works describing topical and oral treatments will be revisited and discussed. The classification, synthesis, and clinical application of these nanomaterials will be also considered in this review article.


Causal discovery using compression-complexity measures.
Causal inference is one of the most fundamental problems across all domains of science. We address the problem of inferring a causal direction from two observed discrete symbolic sequences X and Y. We present a framework which relies on lossless compressors for inferring context-free grammars (CFGs) from sequence pairs and quantifies the extent to which the grammar inferred from one sequence compresses the other sequence. We infer X causes Y if the grammar inferred from X better compresses Y than in the other direction. To put this notion to practice, we propose three models that use the Compression-Complexity Measures (CCMs) - Lempel-Ziv (LZ) complexity and Effort-To-Compress (ETC) to infer CFGs and discover causal directions without demanding temporal structures. We evaluate these models on synthetic and real-world benchmarks and empirically observe performances competitive with current state-of-the-art methods. Lastly, we present two unique applications of the proposed models for causal inference directly from pairs of genome sequences belonging to the SARS-CoV-2 virus. Using numerous sequences, we show that our models capture causal information exchanged between genome sequence pairs, presenting novel opportunities for addressing key issues in sequence analysis to investigate the evolution of virulence and pathogenicity in future applications.


Different faces of the white cataract: a phaco surgeon's perspective.
To develop a system of categorizing white cataracts so that the salient features, as would be pertinent to phaco-emulsification surgery, could be recognized pre-operatively. Patients with total cataracts were carefully evaluated using slit-lamp biomicroscopy and slit-lamp photography. An attempt was made to determine whether pre-operative evaluation could ascertain the presence or absence of features of these cataracts that often cause surgical difficulties, namely, increased intralenticular pressure (ILP), milky cortex and brown nuclear colour. Using a standardized method of biomicroscopic evaluation, it was found that the status of the ILP, cortex and nuclear colour could be determined consistently. Surgical strategies based on such a system of categorization has permitted white cataracts to be consistently managed using phacoemulsification. Systematic pre-operative evaluation and categorization of white cataracts based on the ILP, status of the cortex and nuclear colour can permit proper surgical planning and successful management of such cases with phacoemulsification.


Model-based control of cardiac alternans in Purkinje fibers.
This paper describes a systematic approach to suppressing cardiac alternans in simulated Purkinje fibers using localized current injections. We investigate the controllability and observability of the periodically paced Noble model for different locations of the recording and control electrodes. In particular, we show that the loss of controllability causes the failure of the control approach introduced by Echebarria and Karma [Chaos 12, 923 (2002)] for longer fiber lengths. Furthermore, we explain how the optimal locations for the recording and control electrodes and the timing of the feedback current can be selected, accounting for both linear and nonlinear effects, effectively doubling the length of fibers that can be controlled with previous methods.


Changes in islet cell composition during development of diabetes in Macaca nigra.
The islets of Langerhans in sections from the pancreas tail of Macaca nigra were stained by antiserum to insulin, glucagon, or somatostatin. The area of stained cells per total area of the islets was determined by a computerized photometric method. Insulin of the beta cells occupied 77% of the islet area in nondiabetic (ND) monkeys and decreased to 62% in monkeys in the earliest stages of metabolic deterioration, i.e., hormonally impaired (HI) monkeys. At the later stage of borderline diabetes (BD), monkeys had only 39% of the islet area occupied by insulin and the area was diminished to less than 1% in diabetic (D) monkeys. Islets in HI monkeys had an unusual pattern in which only the beta cells in the periphery of islets were stained. Glucagon in the alpha cells stained 7% of the islet area in ND monkeys, but the area was almost doubled to 13% in HI monkeys; the percentage decreased to about 5% in BD and 3% in D monkeys. Somatostatin accounted for 5% of the islet area in ND monkeys, was slightly greater at 7% in HI monkeys, and decreased to 3% in BD and 2% in D monkeys. Alterations in percentages of secretory cells correlated with several of the metabolic and clinical changes.


Influence of an increase in excise duty on alcohol consumption and its adverse effects.
The excise duty on alcoholic beverages was increased in March 1981, causing the cost of alcohol to rise faster than other prices for the first time in 30 years. For this reason 463 "regular drinkers" in the Lothian region whose drinking habits had been established in 1978-9 were reinterviewed in 1981-2. Overall, their alcohol consumption had fallen by 18% and associated adverse effects by 16%. Heavy drinkers and suspected dependent drinkers both reduced their consumption at least as much as light or moderate drinkers and suffered considerably fewer adverse effects as a result. Increasing the excise duty on alcoholic beverages can therefore be an effective public health measure. Factors related to rising unemployment were responsible for about 20% of the overall reduction in consumption.


Kolaviron attenuates diclofenac-induced nephrotoxicity in male Wistar rats.
The beneficial effects of kolaviron, a natural biflavonoid from the seeds of Garcinia kola, have been attributed to its antioxidant and anti-inflammatory activities. This study was designed to investigate the renoprotective effect of kolaviron in rat model of diclofenac (DFC)-induced acute renal failure. Thirty-five male Wistar rats were divided into 7 groups of 5 rats each as follows: a control group that received propylene glycol orally and treatment groups that received DFC, DFC recovery, DFC followed by kolaviron at 3 different doses, and kolaviron only. DFC-treated rats showed sluggishness, illness, and anorexia. Their urine contained appreciable protein, glucose, and ketone bodies. Histopathological examination of their kidneys revealed profound acute tubular necrosis. DFC treatment significantly increased levels of plasma creatinine, urea, sodium, chloride, potassium ions, and increased renal tissue activities of superoxide dismutase, catalase, levels of malondialdehyde, and hydrogen peroxide. Fractional excretion of sodium and potassium and renal tissue levels of reduced glutathione and prostaglandin E2 (PGE2) decreased significantly in DFC-treated groups. However, kolaviron administration significantly reduced the toxic effect of DFC on PGE2 release; plasma levels of creatinine, urea, glucose, and electrolytes; and significantly attenuated renal tubular and oxidative damages. Furthermore, the effects of DFC administration on food consumption, water intake, urine output and urine protein, glucose, ketone bodies, and electrolytes were significantly attenuated in animals treated with kolaviron. The results suggested that kolaviron ameliorated DFC-induced kidney injury in Wistar rats by decreasing renal oxidative damage and restoration of renal PGE2 release back to the basal levels.


Bring in the genes: genetic-ecophysiological modeling of the adaptive response of trees to environmental change. With application to the annual cycle.
The observation of strong latitudinal clines in the date of bud burst of tree species indicate that populations of these species are genetically adapted to local environmental conditions. Existing phenological models rarely address this clinal variation, so that adaptive responses of tree populations to changes in environmental conditions are not taken into account, e.g., in models on species distributions that use phenological sub-models. This omission of simulating adaptive response in tree models may over- or underestimate the effects of climate change on tree species distributions, as well as the impacts of climate change on tree growth and productivity. Here, we present an approach to model the adaptive response of traits to environmental change based on an integrated process-based eco-physiological and quantitative genetic model of adaptive traits. Thus, the parameter values of phenological traits are expressed in genetic terms (allele effects and-frequencies, number of loci) for individual trees. These individual trees thereby differ in their ability to acquire resources, grow and reproduce as described by the process-based model, leading to differential survival. Differential survival is thus the consequence of both differences in parameters values and their genetic composition. By simulating recombination and dispersal of pollen, the genetic composition of the offspring will differ from that of their parents. Over time, the distribution of both trait values and the frequency of the underlying alleles in the population change as a consequence of changes in environmental drivers leading to adaptation of trees to local environmental conditions. This approach is applied to an individual-tree growth model that includes a phenological model on the annual cycle of trees whose parameters are allowed to adapt. An example of the adaptive response of the onset of the growing season across Europe is presented.


Modulation of islet G-proteins, alpha-glucosidehydrolase inhibition and insulin release stimulated by various secretagogues.
Guanine nucleotide-binding proteins (G-proteins) are known to act as important modulators of insulin release from the islets of Langerhans. We have recently found that the deoxynojirimycin-derivative emiglitate, a recognized inhibitor of intestinal alpha-glucosidehydrolase activity, is a powerful inhibitor of glucose-induced insulin release. With the use of isolated mouse islets the present investigation was performed in a primary attempt to elucidate whether this inhibitory mechanism in some way was linked to the beta-cell G-protein system. Treatment of freshly isolated islets with pertussis toxin (PTX), which is known to inactivate the G (i)-proteins, abolished the inhibitory effect of the alpha(2)-adrenoceptor agonist clonidine on insulin release stimulated by the phosphodiesterase inhibitor IBMX in the presence of the protein kinase C activator TPA and even changed it into an increase. Emiglitate did not display any inhibitory action on insulin release induced by these secretagogues. Similarly, clonidine-induced inhibition of glucose stimulated insulin release was reversed by PTX. However, PTX did not influence the suppressive action of emiglitate on glucose-induced insulin secretion. In contrast, the adenylate cyclase activator forskolin totally abolished the inhibitory effect of emiglitate, but not that of the glucose analogue mannoheptulose, on glucose-induced insulin release. Moreover, the stimulatory effect of forskolin and cholera toxin (CTX) (activator of G (s)-proteins) on the secretion of insulin was markedly enhanced in the presence of emiglitate. In conclusion, our results suggest that the inhibitory effect of emiglitate on glucose-induced insulin release is not directly related to the G(s)-proteins, but most likely exerted solely through the selective suppression of lysosomal aglucosidehydrolase activity, a step in between the proximal and the distal G(i)-proteins, in glucose induced stimulus-secretion mechanisms. Our data also suggests that the inhibitory action of emiglitate on glucose stimulated insulin release can be compensated for by an increased sensitivity of the cyclic AMP-protein kinase A pathway. Hence, emiglitate might indirectly elicit an increased activity of the G(s)-proteins to facilitate the secretory process.


Metabolic tumor imaging using magnetic resonance spectroscopy.
The adaptability and the genomic plasticity of cancer cells, and the interaction between the tumor microenvironment and co-opted stromal cells, coupled with the ability of cancer cells to colonize distant organs, contribute to the frequent intractability of cancer. It is becoming increasingly evident that personalized molecular targeting is necessary for the successful treatment of this multifaceted and complex disease. Noninvasive imaging modalities such as magnetic resonance (MR), positron emission tomography (PET), and single-photon emission computed tomography (SPECT) are filling several important niches in this era of targeted molecular medicine, in applications that span from bench to bedside. In this review we focus on noninvasive magnetic resonance spectroscopy (MRS) and spectroscopic imaging (MRSI) and their roles in future personalized medicine in cancer. Diagnosis, the identification of the most effective treatment, monitoring treatment delivery, and response to treatment are some of the broad areas into which MRS techniques can be integrated to improve treatment outcomes. The development of novel probes for molecular imaging--in combination with a slew of functional imaging capabilities--makes MRS techniques, especially in combination with other imaging modalities, valuable in cancer drug discovery and basic cancer research.


Anatomical and functional reconstruction of the nigrostriatal system in vitro: selective innervation of the striatum by dopaminergic neurons.
To study development of the nigrostriatal pathway in an in vitro model system, organotypic slices obtained from rat pups (P4) and containing the striatum and the cortex were grown together with apposed embryonic (E13.5) mesencephalic blocks according to the static slice culture method of Stoppini et al. (1991; J. Neurosci. Methods 37:173-182). Under these conditions, mesencephalic dopaminergic (DA) fibers rapidly grow through the slice, preferentially its striatal portion. This innervation provides a true synaptic innervation to the striatum, as shown by the presence of DA terminals on striatal neurons. DA fibers are able to exert a functional influence, as seen by their ability to modulate c-Fos expression in striatal neurons in the same way as in vivo. Thus, blockade, under basal conditions, of the effect of spontaneously released dopamine by the D2 receptor antagonist haloperidol leads to the activation of c-Fos expression in the striatum. Furthermore, stimulation of DA release by amphetamine induces striatal c-Fos expression in a D1 receptor-dependent manner. Next, the mechanisms of the selective striatal innervation were examined. Indeed, DA fibers innervated specifically the striatum, avoiding the cortical portion of the slice. This selectivity seems to be specific for DA neurons; no selectivity could be observed when noradrenergic neurons were substituted for DA neurons. Short-term cocultures in a collagen gel of mesencephalic blocks with striatal blocks failed to reveal any oriented outgrowth of DA fibers from the mesencephalon, suggesting that the selective innervation observed in the organotypic slices results from some contact-dependent, presumably adhesive interactions rather than from the presence of some diffusible substance orienting the growth of DA fibers towards the striatum. On the other hand, DA neurons seeded onto striatal slices did not attach selectively onto the striatal portion of the slice, indicating that the putative specific adhesive interactions governing the selective striatal innervation are not the same as those determining the adhesion of the DA neurons. These results show that cocultures of cortex-striatum and mesencephalic slices result in a system that displays a number of the morphological and functional traits of the normal nigrostriatal system and that can be relied on as a good in vitro model of in vivo development.


Evidence for multidrug-resistant cells in human tumor cell populations.
Data are presented from two cell culture systems which support the notion that a multi-drug-resistant phenotype occurs in human tumor cell populations. Human small cell lung cancer cell lines derived from patients in relapse following intensive combination chemotherapy demonstrate broad cross-resistance to nine standard drugs in vitro. However, analysis of [14C]glucosamine-labeled glycoproteins in the small cell lung cancer cell lines failed to identify any consistent association between a specific glycoprotein marker and the drug-resistant phenotype. Evaluation of drug sensitivity of human tumor cells in primary culture (colony-forming assay) has indicated that multidrug-resistant cells may be present in tumor cell populations even in the absence of prior drug therapy. Several features of the multidrug-resistant phenotype, as observed in these human tumor cell populations, differ from those observed in Chinese hamster cell systems. In particular, the variability in patterns of resistance to various agents and in expression of glycoprotein markers suggests that a substantial amount of genetic heterogeneity underlies this phenotype in human tumors.


Efficacy of Eucalyptus cinerea as a Source of Bioactive Compounds for Curative Biocontrol of Crown Gall Caused by Agrobacterium tumefaciens Strain B6.
This research investigated the Eucalyptus cinerea leaves efficiency in the Agrobacterium tumefaciens biocontrol, the causative agent of crown gall. GC-MS analysis of the essential oil (EO) showed that the main components were 1,8-cineole (61%) and camphene (15.13%). Thanks to its polyphenols, flavonoids, quinones, terpenoids, alkaloids, and tannins richness, the EtOAc-F exhibited the most potent antibacterial activity in vitro. Indeed, compared to the other fractions, it has the lowest MIC and MBC values of 0.312 mg/mL and 2.5 mg/mL, respectively. The GC-MS analysis of EtOAc-F confirmed its richness in antibacterial compounds including gallic acid (7.18%), shikimic acid (5.07%), and catechin (3.12%). The time-kill curve assay of EtOAc-F (2.5 mg/mL) showed a potent bactericidal effect after 20 min of direct contact with A. tumefaciens. In planta experiments, gall weights were significantly reduced when EtOAc-F was applied at 0.625 and 2.5 mg/wounds. Besides, the disease reduction rates in gall weight were 95% and 97.5%, respectively. Interestingly, no phytotoxic effect was observed since tomato seeds germinated in the presence of the different concentrations of EtOAc-F. These results suggest that EtOAc-F has a good potential to be a curative biocontrol agent for crown gall disease.


Influence of Immediate Implant Placement and Provisionalization with or without Soft Tissue Augmentation on Hard and Soft Tissues in the Esthetic Zone: A One-Year Retrospective Study.
The purpose of this clinical research was to evaluate peri-implant marginal changes around immediate implants placed either with the application of SCTG or XCM or without soft tissue grafting. A total of 48 patients requiring a single implant-supported restoration in the anterior jaw were selected for inclusion. Three surgical procedures were performed, as follows: type 1 implant with subepithelial connective tissue graft (SCTG), type 1 implant with xenogenic collagen matrix (XCM), and type 1 implant without soft tissue augmentation (NG) (control group). The marginal change of peri-implant soft tissue, facial soft tissue thickness (FSTT), peri-implant health status, esthetics, and patient satisfaction were assessed at one year after surgery. All of the placed implants showed a survival rate of 100%. No significant differences in FSTT were recorded between the SCTG group and the XCM group after treatment (P > 0.05), while the NG group presented a significant difference (P < 0.05). Patients in the NG group lost significantly more in the buccal marginal level than did patients in the SCTG group and those in the XCM group (P < 0.05). The favourable success rate recorded in all groups confirmed immediate tooth replacement as a choice of treatment for a missing anterior single tooth. The NG group presented significant changes of FSTT and buccal marginal level, while XCM constituted a viable alternative to SCTG.


Parafoveal preview effects in reading unspaced text.
In English reading, eye guidance relies heavily on the spaces between words for demarcating word boundaries. In an eye tracking experiment, we examined the impact of removing spaces on parafoveal processing. Using the gaze-contingent boundary paradigm (Rayner, 1975), a high or low frequency preboundary word was followed by a postboundary preview presented either normally (i.e., identical to the postboundary word), or with letters replaced creating an orthographically illegal preview. The spaces between words were either retained or removed. Results replicate previous findings of increased reading times during unspaced reading (Rayner, Fischer, & Pollatsek, 1998) and indicate rather limited evidence for more distributed processing: Observations of processing of the previous word (spill-over effects) or processing of the next word (parafoveal-on-foveal effects) influencing fixation durations on the currently fixated word were limited. Spill-over effects were only observed in the unspaced layout when the postboundary preview was correct, presumably because the orthographically illegal, incorrect preview was visually salient enough to allow for relatively easy word segmentation and therefore more focused processing of the preboundary word. As such, results points toward a system that prefers narrowly focused processing of a single word, at least when means for easy word segmentation are available. (PsycINFO Database Record


The New York Consortium on Membrane Protein Structure (NYCOMPS): a high-throughput platform for structural genomics of integral membrane proteins.
The New York Consortium on Membrane Protein Structure (NYCOMPS) was formed to accelerate the acquisition of structural information on membrane proteins by applying a structural genomics approach. NYCOMPS comprises a bioinformatics group, a centralized facility operating a high-throughput cloning and screening pipeline, a set of associated wet labs that perform high-level protein production and structure determination by x-ray crystallography and NMR, and a set of investigators focused on methods development. In the first three years of operation, the NYCOMPS pipeline has so far produced and screened 7,250 expression constructs for 8,045 target proteins. Approximately 600 of these verified targets were scaled up to levels required for structural studies, so far yielding 24 membrane protein crystals. Here we describe the overall structure of NYCOMPS and provide details on the high-throughput pipeline.


A status report on diet, nutrition and cancer.
Current data imply that a compilation of the nutrient effects strongly suggests that a diet enriched in plant products reduces cancer risk. The recommendation to increase the consumption of plant products is in accord with similar recommendations on diet and other chronic diseases, such as cardiovascular disease and diabetes.


Redefining the DNA-binding domain of human XPA.
Xeroderma pigmentosum complementation group A (XPA) protein plays a critical role in the repair of DNA damage via the nucleotide excision repair (NER) pathway. XPA serves as a scaffold for NER, interacting with several other NER proteins as well as the DNA substrate. The critical importance of XPA is underscored by its association with the most severe clinical phenotypes of the genetic disorder Xeroderma pigmentosum. Many of these disease-associated mutations map to the XPA(98-219) DNA-binding domain (DBD) first reported ~20 years ago. Although multiple solution NMR structures of XPA(98-219) have been determined, the molecular basis for the interaction of this domain with DNA is only poorly characterized. In this report, we demonstrate using a fluorescence anisotropy DNA-binding assay that the previously reported XPA DBD binds DNA with substantially weaker affinity than the full-length protein. In-depth analysis of the XPA sequence suggested that the original DBD construct lacks critical basic charge and helical elements at its C-terminus. Generation and analysis of a series of C-terminal extensions beyond residue 219 yielded a stable, soluble human XPA(98-239) construct that binds to a Y-shaped ssDNA-dsDNA junction and other substrates with the same affinity as the full-length protein. Two-dimensional (15)N-(1)H NMR suggested XPA(98-239) contains the same globular core as XPA98-219 and likely undergoes a conformational change upon binding DNA. Together, our results demonstrate that the XPA DBD should be redefined and that XPA(98-239) is a suitable model to examine the DNA binding activity of human XPA.


Nitric oxide regulates renal cortical cyclooxygenase-2 expression.
We have previously shown that cyclooxygenase-2 (COX-2) is localized to the cortical thick ascending limb of the loop of Henle (cTALH)/macula densa of the rat kidney, and expression increases in response to low-salt diet and/or angiotensin-converting enzyme (ACE) inhibition. Because of the localization of neuronal nitric oxide synthase (nNOS) to macula densa and surrounding cTALH, the present study investigated the role of nitric oxide (NO) in the regulation of COX-2 expression. For in vivo studies, rats were fed a normal diet, low-salt diet or low-salt diet combined with the ACE inhibitor captopril. In each group, one-half of them were treated with the nNOS inhibitors 7-nitroinidazole (7-NI) or S-methyl-thiocitrulline. Both of these NOS inhibitors inhibited increases in COX-2 mRNA and immunoreactive protein in response to low salt and low salt+captopril. For in vitro studies, COX-2 expression was studied in primary cultures of rabbit cTALH cells immunodisssected with Tamm-Horsfall antibody. Basal COX-2 immunoreactivity expression was stimulated by S-nitroso-N-acetyl-penicillamine (SNAP), an NO donor, and intracellular cGMP concentration. The cultured cells expressed immunoreactive nNOS, and 7-NI inhibited basal COX-2 immunoreactivity expression, which could be partially overcome by cGMP. In summary, these studies indicate that NO is a mediator of increased renal cortical COX-2 expression seen in volume depletion and suggest important interactions between the NO and COX-2 systems in the regulation of arteriolar tone and the renin-angiotensin system by the macula densa.


Family History Information Extraction With Neural Attention and an Enhanced Relation-Side Scheme: Algorithm Development and Validation.
Identifying and extracting family history information (FHI) from clinical reports are significant for recognizing disease susceptibility. However, FHI is usually described in a narrative manner within patients' electronic health records, which requires the application of natural language processing technologies to automatically extract such information to provide more comprehensive patient-centered information to physicians. This study aimed to overcome the 2 main challenges observed in previous research focusing on FHI extraction. One is the requirement to develop postprocessing rules to infer the member and side information of family mentions. The other is to efficiently utilize intrasentence and intersentence information to assist FHI extraction. We formulated the task as a sequential labeling problem and propose an enhanced relation-side scheme that encodes the required family member properties to not only eliminate the need for postprocessing rules but also relieve the insufficient training instance issues. Moreover, an attention-based neural network structure was proposed to exploit cross-sentence information to identify FHI and its attributes requiring cross-sentence inference. The dataset released by the 2019 n2c2/OHNLP family history extraction task was used to evaluate the performance of the proposed methods. We started by comparing the performance of the traditional neural sequence models with the ordinary scheme and enhanced scheme. Next, we studied the effectiveness of the proposed attention-enhanced neural networks by comparing their performance with that of the traditional networks. It was observed that, with the enhanced scheme, the recall of the neural network can be improved, leading to an increase in the F score of 0.024. The proposed neural attention mechanism enhanced both the recall and precision and resulted in an improved F score of 0.807, which was ranked fourth in the shared task. We presented an attention-based neural network along with an enhanced tag scheme that enables the neural network model to learn and interpret the implicit relationship and side information of the recognized family members across sentences without relying on heuristic rules.


Clustering of Dietary Patterns and Lifestyles Among Spanish Children in the EsNuPI Study †.
Dietary patterns (DPs) are known to be tied to lifestyle behaviors. Understanding DPs and their relationships with lifestyle factors can help to prevent children from engaging in unhealthy dietary practices. We aimed to describe DPs in Spanish children aged 1 to <10 years and to examine their associations with sociodemographic and lifestyle variables. The consumption of toddler and young children milk formulas, enriched and fortified milk within the Spanish pediatric population is increasing, and there is a lack of evidence whether the consumption of this type of milk is causing an impact on nutrient intakes and if they are helping to reach the nutrient recommendations. Within the Nutritional Study in the Spanish Pediatric Population (EsNuPI), we considered two study cohorts and three different age groups in three year-intervals in each of them. The study cohort included 740 children in a representative sample of the urban non-vegan Spanish population and 772 children in a convenience cohort of adapted milk consumers (AMS) (including follow-on formula, toddler's milk, growing up milk, and fortified and enriched milks) who provided information about sociodemographics, lifestyle, and dietary habits; a food frequency questionnaire was used for the latter. Principal component analysis was performed to identify DPs from 18 food groups. Food groups and sociodemographic/lifestyle variables were combined through a hierarchical cluster algorithm. Three DPs predominated in every age group and study sample: a palatable energy-dense food dietary pattern, and two Mediterranean-like DPs. However, children from the AMS showed a predominant dietary pattern markedly related to the Mediterranean diet, with high consumption of cereals, fruits and vegetables, as well as milk and dairy products. The age of children and certain lifestyle factors, namely level of physical activity, parental education, and household income, correlated closely with the dietary clusters. Thus, the findings provide insight into designing lifestyle interventions that could reverse the appearance of unhealthy DPs in the Spanish child population.


Tau protects microtubules in the axon from severing by katanin.
Microtubules in the axon are more resistant to severing by katanin than microtubules elsewhere in the neuron. We have hypothesized that this is because of the presence of tau on axonal microtubules. When katanin is overexpressed in fibroblasts, the microtubules are severed into short pieces, but this phenomenon is suppressed by the coexpression of tau. Protection against severing is also afforded by microtubule-associated protein 2 (MAP2), which has a tau-like microtubule-binding domain, but not by MAP1b, which has a different microtubule-binding domain. The microtubule-binding domain of tau is required for the protection, but within itself, provides less protection than the entire molecule. When tau (but not MAP2 or MAP1b) is experimentally depleted from neurons, the microtubules in the axon lose their characteristic resistance to katanin. These results, which validate our hypothesis, also suggest a potential explanation for why axonal microtubules deteriorate in neuropathies involving the dissociation of tau from the microtubules.


Alignment of synaptic vesicle macromolecules with the macromolecules in active zone material that direct vesicle docking.
Synaptic vesicles dock at active zones on the presynaptic plasma membrane of a neuron's axon terminals as a precondition for fusing with the membrane and releasing their neurotransmitter to mediate synaptic impulse transmission. Typically, docked vesicles are next to aggregates of plasma membrane-bound macromolecules called active zone material (AZM). Electron tomography on tissue sections from fixed and stained axon terminals of active and resting frog neuromuscular junctions has led to the conclusion that undocked vesicles are directed to and held at the docking sites by the successive formation of stable connections between vesicle membrane proteins and proteins in different classes of AZM macromolecules. Using the same nanometer scale 3D imaging technology on appropriately stained frog neuromuscular junctions, we found that ∼10% of a vesicle's luminal volume is occupied by a radial assembly of elongate macromolecules attached by narrow projections, nubs, to the vesicle membrane at ∼25 sites. The assembly's chiral, bilateral shape is nearly the same vesicle to vesicle, and nubs, at their sites of connection to the vesicle membrane, are linked to macromolecules that span the membrane. For docked vesicles, the orientation of the assembly's shape relative to the AZM and the presynaptic membrane is the same vesicle to vesicle, whereas for undocked vesicles it is not. The connection sites of most nubs on the membrane of docked vesicles are paired with the connection sites of the different classes of AZM macromolecules that regulate docking, and the membrane spanning macromolecules linked to these nubs are also attached to the AZM macromolecules. We conclude that the luminal assembly of macromolecules anchors in a particular arrangement vesicle membrane macromolecules, which contain the proteins that connect the vesicles to AZM macromolecules during docking. Undocked vesicles must move in a way that aligns this arrangement with the AZM macromolecules for docking to proceed.


Synthesis of fructose bound Fe(iii) integrated carbon dots as a robust turn-off detection sensor for chlortoluron.
A simple, sensitive, and robust fluorescent sensor for chlortoluron detection has been developed. Fluorescent carbon dots were synthesized using ethylene diamine and fructose via a hydrothermal protocol. The molecular interaction between fructose carbon dots and Fe(iii) resulted in a fluorescent metastable state exhibiting remarkable fluorescence quenching at λem of 454 nm and interestingly, further quenching occurred upon the addition of chlortoluron. The quenching in the fluorescence intensity of CDF-Fe(iii) towards chlortoluron occurred in the concentration range of 0.2-5.0 μg mL-1 where the limit of detection was found to be 0.0467 μg mL-1, the limit of quantification was 0.14 μg mL-1, and the relative standard deviation was 0.568%. The selective and specific recognitive nature of the Fe(iii) integrated fructose bound carbon dots towards the chlortoluron make it a suitable sensor for real sample applications. The proposed strategy was applied for the determination of chlortoluron in soil, water, and wheat samples with recoveries in the range of 95% to 104.3%.


Are tobacco subsidies a misuse of public funds?
The European Union spends about 1000m ecu (800m pounds, $1240m) a year subsidizing tobacco production but only about 1.55m ecu (1.2m pounds, $1.85m) on smoking prevention. The subsidies, part of the common agricultural policy, were originally intended to encourage farmers to grow commercially valued varieties of tobacco and thus reduce imports. But they also aimed to guarantee farmers' income, a goal in direct conflict with the first. The policy has failed to adapt production to demand or reduce imports, since most tobacco grown in the union has little commercial value. Reforms introduced in 1992 have had limited impact on expenditure, and data produced as a result of the reforms show that it would be much cheaper to give farmers direct income support than to subsidize them growing tobacco. Tobacco subsidies should be abolished and more should be spent on smoking prevention.


Are floating toes associated with lifestyle in children? A cross-sectional study.
Floating toes are a condition and deformity in which some of the toes are afloat. Many functional impairments in floating toes have been previously studied lately and several factors related to floating toes have also been reported. However, no reports have considered the relationship between lifestyle and floating toes among children. The purpose of this study was to reveal the prevalence of floating toes among school children and reveal its relationship with lifestyle. In total, 138 young male baseball players were recruited. Lifestyle was evaluated by using a questionnaire and chosen whether the main lifestyle was Japanese or Western, if the bedding was futons or beds, and if the toilet was Japanese style (a squat toilet) or Western style. Floating toes were defined as toes that were not in contact with the mat. Ankle dorsiflexion in the knee-flexed and knee-extended positions was measured in a weight-bearing position. The relationship between the floating toes and lifestyles, and the comparison of ankle dorsiflexion range of motion between the lifestyles were statistically analyzed. Players living in a Western style showed a significantly higher prevalence of floating toes on both feet compared with the players living in a Japanese style (throwing side; 39% vs. 19%, p = 0.04, and non-throwing side; 43% vs. 19%, p = 0.01). Players living in a Western style with beds showed a significantly smaller range of motion on both sides of ankle dorsiflexion in the knee-flexed position compared with those who were not (throwing side; 37.2 ± 5.7° vs. 39.0 ± 6.6°, p = 0.04, and non-throwing side; 36.8 ± 5.8° vs. 38.6 ± 6.1°, p = 0.04). Children mainly living in a Western lifestyle showed a significantly higher prevalence of floating toes on both feet compared to those mainly living in a Japanese lifestyle. The prevalence of floating toes may be related to lifestyles among children. The study was approved by the institutional review board of the Waseda University Graduate School of Sport Sciences (IRB number 2021-185).


Monthly sulphadoxine-pyrimethamine combination versus daily proguanil for malaria chemoprophylaxis in sickle cell disease: a randomized controlled study at the Jos University Teaching Hospital.
Malaria carries a high case fatality among patients with sickle cell disease. In Jos University Teaching Hospital, at the time of this study, the use of Proguanil was the acceptable mode of chemoprophylaxis for preventing malaria in these patients. Intermittent Preventive Treatment (IPT) with Sulphadoxine-Pyrimethamine [SP] has shown great potential for reducing the prevalence of malaria and anaemia among pregnant women, infants and travellers. We hypothesised that monthly SP was superior to daily Proguanil in reducing malaria parasitaemia, clinical malaria attacks and sickle cell crises in such patients. To assess the efficacy and affordability of monthly SP versus daily Proguanil for malaria chemoprophylaxis in patients attending Sickle Cell Clinic at Jos University Teaching Hospital, Plateau State, Nigeria. One hundred and fifty four patients [114 children and 40 adults] with Sickle Cell Disease in their steady state were randomized to monthly SP or daily Proguanil for malaria chemoprophylaxis. Active detection of malaria parasite in the peripheral blood and packed cell volumes were done at each monthly visit to the clinic over a period of three months. The primary outcome measure was the proportion of patients with malaria parasite in the peripheral blood at the end of 3 months. The secondary outcome measures included episodes of clinical malaria attacks, frequency and type of sickle cell crises and adverse effects of the medication. Ninety four percent [72/77] of patients in the SP group and 91% [70/77] in the Proguanil group respectively completed three months of follow up. SP reduced the prevalence of malaria parasitaemia by 25% [(14%) 10/72] compared to 6.4% [(30%) 21/70] in the proguanil group. [X2 54; p = 0.01]. Seventeen percent [12/72] of the patients receiving monthly SP had malaria attacks compared to 57% [40/70] on prophylaxis with Proguanil. [X2 =25; p< 0.0003]. Thirty three percent [24/72] of the patients receiving SP had at least an episode of bone pain crises compared to 69% [48/70] of the patients receiving Proguanil. [X2 =17.6; p<0.0001]. SP was 8 times cheaper than Proguanil. Monthly chemoprophylaxis with SP was more efficacious than daily Proguanil in reducing the prevalence of asymptomatic malaria parasitaemia, clinical malaria attack and sickle cell crises in patients with sickle cell disease. SP was 8 times cheaper than Proguanil. No significant side effect was recorded in both groups. The current practice of routinely prescribing daily Proguanil to SCD patients for malaria chemoprophylaxis needs to be reviewed.


The genetics of alcohol intake and of alcohol dependence.
Because alcohol has multiple dose-dependent consequences, it is important to understand the causes of individual variation in the amount of alcohol used. The aims of this study were to assess the long-term repeatability and genetic or environmental causes of variation in alcohol intake and to estimate the degree of overlap with causes of susceptibility to alcohol dependence. Data were used from three studies conducted between 1980 and 1995 on volunteer adult male and female Australian twin subjects. In each study, alcohol intake was reported both as quantity x frequency and as past-week data. Repeatability was calculated as correlations between occasions and between measures, and the effects of genes and environment were estimated by multivariate model fitting to the twin pair repeated measures of alcohol use. Relationships between mean alcohol use and the lifetime history of DSM-III-R alcohol dependence were tested by bivariate model fitting. Repeatability of the alcohol intake measures was between 0.54 and 0.85, with the highest repeatability between measures within study and the lowest repeatability between the first and last studies. Reported alcohol consumption was mainly affected by genetic factors affecting all times of study and by nonshared environmental factors (including measurement error) unique to each time of study. Genes that affect alcohol intake do affect alcohol dependence, but genetic effects unique to dependence are also significant; environmental effects are largely unique to either intake and dependence. Nearly all the repeatable component of variation in alcohol intake is due to genetic effects. Genes affecting intake also affect dependence risk, but there are other genes that affect dependence alone. Studies aiming to identify genes that affect alcohol use disorders need to test loci and candidate genes against both phenotypes.


Targeting of (D)MinC/MinD and (D)MinC/DicB complexes to septal rings in Escherichia coli suggests a multistep mechanism for MinC-mediated destruction of nascent FtsZ rings.
The MinC protein is an important determinant of septal ring positioning in Escherichia coli. The N-terminal domain ((Z)MinC) suppresses septal ring formation by interfering with FtsZ polymerization, whereas the C-terminal domain ((D)MinC) is required for dimerization as well as for interaction with the MinD protein. MinD oscillates between the membrane of both cell halves in a MinE-dependent fashion. MinC oscillates along with MinD such that the time-integrated concentration of (Z)MinC at the membrane is minimal, and hence the stability of FtsZ polymers is maximal, at the cell center. MinC is cytoplasmic and fails to block FtsZ assembly in the absence of MinD, indicating that recruitment of MinC by MinD to the membrane enhances (Z)MinC function. Here, we present evidence that the binding of (D)MinC to MinD endows the MinC/MinD complex with a more specific affinity for a septal ring-associated target in vivo. Thus, MinD does not merely attract MinC to the membrane but also aids MinC in specifically binding to, or in close proximity to, the substrate of its (Z)MinC domain. MinC-mediated division inhibition can also be activated in a MinD-independent fashion by the DicB protein of cryptic prophage Kim. DicB shows little homology to MinD, and how it stimulates MinC function has been unclear. Similar to the results obtained with MinD, we find that DicB interacts directly with (D)MinC, that the (D)MinC/DicB complex has a high affinity for some septal ring target(s), and that MinC/DicB interferes with the assembly and/or integrity of FtsZ rings in vivo. The results suggest a multistep mechanism for the activation of MinC-mediated division inhibition by either MinD or DicB and further expand the number of properties that can be ascribed to the Min proteins.


A Rule-Based Data Quality Assessment System for Electronic Health Record Data.
Rule-based data quality assessment in health care facilities was explored through compilation, implementation, and evaluation of 63,397 data quality rules in a single-center case study to assess the ability of rules-based data quality assessment to identify data errors of importance to physicians and system owners. We applied a design science framework to design, demonstrate, test, and evaluate a scalable framework with which data quality rules can be managed and used in health care facilities for data quality assessment and monitoring. We identified 63,397 rules partitioned into 28 logic templates. A total of 819,683 discrepancies were identified by 4.5% of the rules. Nine out of 11 participating clinical and operational leaders indicated that the rules identified data quality problems and articulated next steps that they wanted to take based on the reported information. The combined rule template and knowledge table approach makes governance and maintenance of otherwise large rule sets manageable. Identified challenges to rule-based data quality monitoring included the lack of curated and maintained knowledge sources relevant to data error detection and lack of organizational resources to support clinical and operational leaders with investigation and characterization of data errors and pursuit of corrective and preventative actions. Limitations of our study included implementation within a single center and dependence of the results on the implemented rule set. This study demonstrates a scalable framework (up to 63,397 rules) with which data quality rules can be implemented and managed in health care facilities to identify data errors. The data quality problems identified at the implementation site were important enough to prompt action requests from clinical and operational leaders.


The tumor suppressor gene HIC1 (hypermethylated in cancer 1) is a sequence-specific transcriptional repressor: definition of its consensus binding sequence and analysis of its DNA binding and repressive properties.
HIC1 (hypermethylated in cancer 1) is a tumor suppressor gene located at chromosome 17p13.3, a region frequently hypermethylated or deleted in human tumors and in a contiguous-gene syndrome, the Miller-Dieker syndrome. HIC1 is a transcriptional repressor containing five Krüppel-like C(2)H(2) zinc fingers and an N-terminal dimerization and autonomous repression domain called BTB/POZ. Although some of the HIC1 transcriptional repression mechanisms have been recently deciphered, target genes are still to be discovered. In this study, we determined the consensus binding sequence for HIC1 and investigated its DNA binding properties. Using a selection and amplification of binding sites technique, we identified the sequence 5'-(C)/(G)NG(C)/(G)GGGCA(C)/(A) CC-3' as an optimal binding site. In silico and functional analyses fully validated this consensus and highlighted a GGCA core motif bound by zinc fingers 3 and 4. The BTB/POZ domain inhibits the binding of HIC1 to a single site but mediates cooperative binding to a probe containing five concatemerized binding sites, a property shared by other BTB/POZ proteins. Finally, full-length HIC1 proteins transiently expressed in RK13 cells and more importantly, endogenous HIC1 proteins from the DAOY medulloblastoma cell line, repress the transcription of a reporter gene through their direct binding to these sites, as confirmed by chromatin immunoprecipitation experiments. The definition of the HIC1-specific DNA binding sequence as well as the requirement for multiple sites for optimal binding of the full-length protein are mandatory prerequisites for the identification and analyses of bona fide HIC1 target genes.


A freeze-stable formulation for DTwP and DTaP vaccines.
Inadvertent vaccine freezing often occurs in the cold chain and may cause damage to freeze‑sensitive vaccines. Liquid vaccines that contain aluminum salt adjuvants are particularly vulnerable. Polyol cryoprotective excipients have been shown to prevent freeze damage to hepatitis B vaccine. In this study, we examined the freeze-protective effect of propylene glycol on diphtheria-tetanus-pertussis-whole-cell (DTwP) and acellular (DTaP) vaccines. Pilot lots of DTwP and DTaP formulated with 7.5% propylene glycol underwent 3 freeze-thaw treatments. The addition of propylene glycol had no impact on pH, particle size distribution, or potency of the vaccines prior to freeze-thaw treatment; the only change noted was an increase in osmolality. The potencies and the physical properties of the vaccines containing cryoprotectant were maintained after freeze-thawing and for 3 months in accelerated stability studies. The results from this study indicate that formulating vaccines with propylene glycol can protect diphtheria-tetanus-pertussis vaccines against freeze damages.


Immune cells in the retina and choroid: Two different tissue environments that require different defenses and surveillance.
In the eye immune defenses must take place in a plethora of differing microenvironments ranging from the corneal and conjunctival epithelia facing the external environment to the pigmented connective tissue of the uveal tract containing smooth muscle, blood vessels and peripheral nerves to the innermost and highly protected neural retina. The extravascular environment of the neural retina, like the brain parenchyma, is stringently controlled to maintain conditions required for neural transmission. The unique physiological nature of the neural retina can be attributed to the blood retinal barriers (BRB) of the retinal vasculature and the retinal pigment epithelium, which both tightly regulate the transport of small molecules and restrict passage of cells and macromolecules from the circulation into the retina in a similar fashion to the blood brain barrier (BBB). The extracellular environment of the neural retina differs markedly from that of the highly vascular, loose connective tissue of the choroid, which lies outside the BRB. The choroid hosts a variety of immune cell types, including macrophages, dendritic cells (DCs) and mast cells. This is in marked contrast to the neural parenchyma of the retina, which is populated almost solely by microglia. This review will describe the current understanding of the distribution, phenotype and physiological role of ocular immune cells behind or inside the blood-retinal barriers and those in closely juxtaposed tissues outside the barrier. The nature and function of these immune cells can profoundly influence retinal homeostasis and lead to disordered immune function that can lead to vision loss.


17q21.31 microdeletion syndrome: further expanding the clinical phenotype.
Microdeletions of the 17q21.31 region are associated with hypotonia, oromotor dyspraxia, an apparently characteristic face, moderate learning disability and have an estimated prevalence of approximately 1 in 16,000. Here we report 3 individuals who extend further the phenotypic spectrum observed with microdeletions of the 17q21.31 region. They all have learning disability, hypotonia, and craniofacial dysmorphism in keeping with previous reported cases. One case has iris-choroid coloboma and partial situs inversus, 2 features that are newly recorded phenotype abnormalities. These deletions were detected from a cohort of 600 individuals with learning disability and congenital anomalies, reflecting that 17q21.31 microdeletions are a common finding in such cases. FISH analysis demonstrated that each of the deletions occurred as de novo events. The deleted region in our cases encompasses the previously defined critical region for 17q21.31, and includes CRHR1 and MAPT, putative candidate genes for the 17q21.31 phenotype. The 17q21.31 microdeletion phenotype is perhaps more variable than previously described despite haploinsufficiency for the same genes in many cases.


Intra-articular Histone Deacetylase Inhibitor Microcarrier Delivery to Reduce Osteoarthritis.
The histone deacetylase inhibitor (HDACi) was a milestone in the treatment of refractory T-cell lymphoma. However, the beneficial effects of HDACi have not been appreciated in osteoarthritis (OA). Herein, we implemented a microcarrier system because of the outstanding advantages of controlled and sustained release, biodegradability, and biocompatibility. The poly(d,l-lactide-co-glycolide) (PLGA) microcapsules have a regulated and sustained release profile with a reduced initial burst release, which can improve the encapsulation efficiency of the Chidamide. The emulsion solvent evaporation strategy was used to encapsulate Chidamide in PLGA microcapsules. The encapsulation of Chidamide was established by UV-vis spectra and scanning electron microscopy. Additionally, the inhibition of Tnnt3 and immune stimulation by Chidamide helped to inhibit cartilage destruction and prevent articular cartilage degeneration. Based on the results, the Chidamide in PLGA microcapsules provides a transformative therapeutic strategy for the treatment of osteoarthritis patients to relieve symptoms and protect against cartilage degeneration.


Determination of the fiber content of enteral feedings.
The importance of dietary fiber in maintaining optimum bowel function has been shown in feeding studies, and many enteral formulas now contain dietary fiber, usually as soy polysaccharide (SP). Generally only total dietary fiber (TDF) values are given on the label or in product literature, with no indication of insoluble dietary fiber (IDF) and soluble dietary fiber (SDF) content. Often product literature highlights the amount of SP contained in the formula, not the TDF. The method used to determine TDF content usually is not specified, making it impossible to compare products on a TDF basis. We compared the dietary fiber content of commercially available enteral formulas objectively to help clinicians make informed choices when selecting fiber-containing formulas. Fiber-containing formulas by suppliers were freeze-dried and total, water-soluble, and water-insoluble dietary fiber measured by the Association of Official Analytical Chemists (AOAC) approved method. TDF per 1000 mL ranged from 4.0 to 9.9 g for blenderized formulas and from 7.2 to 20.3 g in formulas containing SP. IDF values per 1000 mL ranged from 2.0 to 5.4 g for blenderized product, and the soy formulas ranged from 5.5 to 16.9 g. SDF values per 1000 mL ranged from 2.0 to 4.5 g in blenderized products, and in soy formulas SDF ranged from 1.7 to 3.8 g. Differences in physiological effects of fiber-containing enteral formulas may be because of differences in fiber composition.


Application of parametric survival analysis to women patients with breast cancer at Jimma University Medical Center.
Public health systems in both industrialized and undeveloped countries continue to struggle with the worldwide problem of breast cancer. In sub-Saharan African countries, notably Ethiopia, it is the form of cancer that strikes women the most commonly. Despite the extreme difficulties, the causes of mortality in Ethiopia have not yet been identified. In addition, little study has been done in this area. Therefore, the major objective of this analysis was to pinpoint the factors that were most responsible for the decreased life expectancy of breast cancer patients at the University of Jimma Medical Center. 552 women who had been treated for breast cancer at Jimma University Medical Center between October 2018 and December 2022 were included in this study, which used a retrospective cohort study design and five-year follow-up data. The most frequent and widely used test for comparing the probability of survival curves between several categorical independent variables was the log-rank test. Next, semi-parametric methods for multivariable analysis using the Cox proportional hazards model were used. Furthermore, a parametric strategy that includes fully parametric survival models better achieves the goal of the analysis. Among covariate, age of patient (ϕ = 254.06; 95% CI (3.95, 7.13), P-value = 0.000), patient live in urban (ϕ = 0.84; 95% CI (-0.35,-0.00), P-value = 0.047), preexisting comorbidity (ϕ = 2.46; 95% CI (0.39, 1.41), P-value = 0.001), overweight women cancer patient (ϕ = 0.05; 95% CI(-4.41,-1.57), P-value = 0.000, positive Axillary Node status cancer patient (ϕ = 0.04; 95% CI(-4.45,-1.88), P-value = 0.000), both surgery and chemotropic baseline treatment patient (ϕ = 0.53; 95% CI(-1.12,-0.16), P-value = 0.009) significantly affected the survival of women breast cancer. Age of breast cancer patient, patient education level, place of residence, marital status, pre-existing comorbidity, axillary node status, estrogen receptor, tumor size, body mass index at diagnosis, stage of cancer, and baseline treatment were found to have a significant effect on time to survive for women with breast cancer at the University of Jimma Medical Center, Oromia region, Ethiopia. However, the covariate histologic grade, number of positive lymph nodes involved, and type of hormone used were insignificant to the survival of breast cancer patients.


Acyl-CoA-Binding Domain-Containing 3 (ACBD3; PAP7; GCP60): A Multi-Functional Membrane Domain Organizer.
Acyl-CoA-binding domain-containing 3 (ACBD3) is a multi-functional scaffolding protein, which has been associated with a diverse array of cellular functions, including steroidogenesis, embryogenesis, neurogenesis, Huntington's disease (HD), membrane trafficking, and viral/bacterial proliferation in infected host cells. In this review, we aim to give a timely overview of recent findings on this protein, including its emerging role in membrane domain organization at the Golgi and the mitochondria. We hope that this review provides readers with useful insights on how ACBD3 may contribute to membrane domain organization along the secretory pathway and on the cytoplasmic surface of intracellular organelles, which influence many important physiological and pathophysiological processes in mammalian cells.


Pathologic shear triggers shedding of vascular receptors: a novel mechanism for down-regulation of platelet glycoprotein VI in stenosed coronary vessels.
Ligand-induced ectodomain shedding of glycoprotein VI (GPVI) is a metalloproteinase-dependent event. We examined whether shear force, in the absence of GPVI ligand, was sufficient to induce shedding of GPVI. Human-citrated platelet-rich plasma or washed platelets were subjected to increasing shear rates in a cone-plate viscometer, and levels of intact and cleaved GPVI were examined by Western blot and ELISA. Pathophysiologic shear rates (3000-10 000 seconds(-1)) induced platelet aggregation and metalloproteinase-dependent appearance of soluble GPVI ectodomain, and GPVI platelet remnant. Shedding of GPVI continued after transient exposure to shear. Blockade of α(IIb)β(3), GPIbα, or intracellular signaling inhibited shear-induced platelet aggregation but minimally affected shear-induced shedding of GPVI. Shear-induced GPVI shedding also occurred in platelet-rich plasma or washed platelets isolated from a von Willebrand disease type 3 patient with no detectable VWF, implying that shear-induced activation of platelet metalloproteinases can occur in the absence of GPVI and GPIbα ligands. Significantly elevated levels of sGPVI were observed in 10 patients with stable angina pectoris, with well-defined single vessel coronary artery disease and mean intracoronary shear estimates at 2935 seconds(-1) (peak shear, 19 224 seconds(-1)). Loss of GPVI in platelets exposed to shear has potential implications for the stability of a forming thrombus at arterial shear rates.


[Unilateral spatial neglect].
Unilateral spatial neglect is a common disorder following focal right hemisphere damage, characterised by a failure to perceive and orient towards stimuli located on the left side of space. These deficits are not attributable to primary sensory or motor loss, but involve a neuropsychological disturbance in the representation of space (corporeal and extracorporeal) and in the mechanisms of attention. A major theoretical interest stems from observations that stimuli that are not consciously detected by the patient may nonetheless be processed by intact brain areas, despite unawareness. Unilateral spatial neglect may lead to poor functional recovery and remains difficult to treat using current rehabilitation procedures.


Apoptotic anticancer effect of alvaradoin E isolated from Alvaradoa haitiensis.
Two anthracenone C-glycosides, alvaradoins E and F, isolated from the leaves of Alvaradoa haitiensis Urb. (Simaroubaceae), were found to have potent inhibitory activities with cultured cancer cells. Using the in vivo hollow fiber model, these compounds demonstrated significant growth inhibition at the i.p. site when tested with KB, LNCaP, and Col2 cells. To determine if these anthracenone C-glycosides mediated anticancer activity through an apoptotic pathway, a series of assays were performed with the 10S isomeric compound, alvaradoin E. With a DAPI assay, treatment of LNCaP cells with alvaradoin E at concentrations of 0.4, 2, 10, or 50 microM for 24 or 48 h showed chromatin condensation, a morphological characteristic of apoptosis. Mitochondrial membrane potential, analyzed with a DiOC6 uptake assay, showed that treatment of LNCaP cells with 0.07, 0.14, 0.28, 0.56, 0.86, and 1.12 microM alvaradoin E for 12 h caused dose-dependent membrane depolarization, another indication of early apoptosis. Also, with an annexin V-FITC assay system, treatment of HL-60 cells with 0.07 microM alvaradoin E for 24 h increased annexin V-FITC binding from 3 to 25.9% (8.6-fold). Finally, with the TUNEL assay system, treatment of HL-60 cells with 1.12 microM alvaradoin E for 32 h increased FITC-dUTP binding from 1.2 to 12.1% (10-fold). These data suggest alvaradoin E is an effective anticancer agent that induces apoptosis. Additional studies to establish clinical utility should be of interest.


Comprehensive Molecular Characterization of Gallbladder Carcinoma and Potential Targets for Intervention.
Gallbladder carcinoma (GBC) is an uncommon and aggressive disease, which remains poorly defined at a molecular level. Here, we aimed to characterize the molecular landscape of GBC and identify markers with potential prognostic and therapeutic implications. GBC samples were analyzed using the MSK-IMPACT (Memorial Sloan Kettering-Integrated Mutation Profiling of Actionable Cancer Targets) platform (targeted NGS assay that analyzes 505 cancer-associated genes). Variants with therapeutic implications were identified using OncoKB database. The associations between recurrent genetic alterations and clinicopathologic characteristics (Fisher exact tests) or overall survival (univariate Cox regression) were evaluated. P values were adjusted for multiple testing. Overall, 244 samples (57% primary tumors and 43% metastases) from 233 patients were studied (85% adenocarcinomas, 10% carcinomas with squamous differentiation, and 5% neuroendocrine carcinomas). The most common oncogenic molecular alterations appeared in the cell cycle (TP53 63% and CDKN2A 21%) and RTK_RAS pathways (ERBB2 15% and KRAS 11%). No recurrent structural variants were identified. There were no differences in the molecular landscape of primary and metastasis samples. Variants in SMAD4 and STK11 independently associated with reduced survival in patients with metastatic disease. Alterations considered clinically actionable in GBC or other solid tumor types (e.g., NTRK1 fusions or oncogenic variants in ERBB2, PIK3CA, or BRCA1/2) were identified in 35% of patients; 18% of patients with metastatic disease were treated off-label or enrolled in a clinical trial based on molecular findings. GBC is a genetically diverse malignancy. This large-scale genomic analysis revealed alterations with potential prognostic and therapeutic implications and provides guidance for the development of targeted therapies.


Eruptive melanocytic nevi during etanercept treatment in an ankylosing spondylitis patient: Case report.
Eruptive melanocytic nevi (EMN) are rare multiple melanocytic lesions that rare and associated with dermatological and systemic diseases. Drug induced EMN is also reported with the use of biological or nonbiological chemotherapeutics, immunosuppressive agents, and melanocyte stimulators. In recent years, the increasing use of biological drugs resulted in an increased reports of EMN induced by these drugs. The objective of this abstract is reporting EMN in a patient receiving etanercept treatment. In addition, we also present a literature review of the previously cases with anti-tumor necrosis factor-α biological agents.


Genetics of coronary artery disease: an update.
In 2007, the first genetic risk variant, 9p21, was simultaneously discovered by two independent groups. 9p21 increases the risk of coronary artery disease in individuals with premature heart disease by twofold, and in the overall population the heterozygote is associated with a 25% increased risk and the homozygote with a 50% increased risk. It is of note that the risk mediated by 9p21 is independent of known risk factors. Since then, with the development of new technologies and the international consortium of CARDIoGRAM, there is now a total of 50 genetic risk variants confirmed and replicated for CAD. Of these 50, 35 mediate their risk by unknown mechanisms, indicating that the pathogenesis of atherosclerosis and myocardial infarction is due to additional factors as yet unknown. The role of genetic risk factors in the management of CAD is yet to be determined. Since many of them are independent of known risk factors, the genetic risk will in the future have to be incorporated into the guidelines, which recommend the target level of plasma LDL-C to be achieved based on the number of risk factors.


NOP14 promotes proliferation and metastasis of pancreatic cancer cells.
Pancreatic cancer (PC) is one of the most malignant cancers worldwide. We describe a novel gene, NOP14, which plays significant roles in PC cell proliferation and migration. Inhibition or overexpression of NOP14 expression in PC cells can reduce or promote motility, proliferation and metastatic capacity in vivo. In parallel, we observed changes in proteins related to migration, such as E-cadherin, vimentin, MMP9, RhoA and p53, along with proteins involved in proliferation, such as MAPK3 and CDK2. Taken together, our study provides new evidence for NOP14 in regulating PC cell proliferation and migration, and may provide new insights for clinical diagnosis and therapy.


OsEDR1 negatively regulates rice bacterial resistance via activation of ethylene biosynthesis.
Rice OsEDR1 is a sequence ortholog of Arabidopsis EDR1. However, its molecular function is unknown. We show here that OsEDR1-suppressing/knockout (KO) plants, which developed spontaneous lesions on the leaves, have enhanced resistance to Xanthomonas oryzae pv. oryzae (Xoo) causing bacterial blight disease. This resistance was associated with increased accumulation of salicylic acid (SA) and jasmonic acid (JA), induced expression of SA- and JA-related genes and suppressed accumulation of 1-aminocyclopropane-1-carboxylic acid (ACC), the direct precursor of ethylene, and expression of ethylene-related genes. OsEDR1-KO plants also showed suppressed production of ethylene. Knockout of OsEDR1 suppressed the ACC synthase (ACS) gene family, which encodes the rate-limiting enzymes of ethylene biosynthesis by catalysing the formation of ACC. The lesion phenotype and enhanced bacterial resistance of the OsEDR1-KO plants was partly complemented by the treatment with ACC. ACC treatment was associated with decreased SA and JA biosynthesis in OsEDR1-KO plants. In contrast, aminoethoxyvinylglycine, the inhibitor of ethylene biosynthesis, promoted expression of SA and JA synthesis-related genes in OsEDR1-KO plants. These results suggest that ethylene is a negative signalling molecule in rice bacterial resistance. In the rice-Xoo interaction, OsEDR1 transcriptionally promotes the synthesis of ethylene that, in turn, suppresses SA- and JA-associated defence signalling.


TRPC3-Nox2 complex mediates doxorubicin-induced myocardial atrophy.
Myocardial atrophy is a wasting of cardiac muscle due to hemodynamic unloading. Doxorubicin is a highly effective anticancer agent but also induces myocardial atrophy through a largely unknown mechanism. Here, we demonstrate that inhibiting transient receptor potential canonical 3 (TRPC3) channels abolishes doxorubicin-induced myocardial atrophy in mice. Doxorubicin increased production of ROS in rodent cardiomyocytes through hypoxic stress-mediated upregulation of NADPH oxidase 2 (Nox2), which formed a stable complex with TRPC3. Cardiomyocyte-specific expression of TRPC3 C-terminal minipeptide inhibited TRPC3-Nox2 coupling and suppressed doxorubicin-induced reduction of myocardial cell size and left ventricular (LV) dysfunction, along with its upregulation of Nox2 and oxidative stress, without reducing hypoxic stress. Voluntary exercise, an effective treatment to prevent doxorubicin-induced cardiotoxicity, also downregulated the TRPC3-Nox2 complex and promoted volume load-induced LV compliance, as demonstrated in TRPC3-deficient hearts. These results illustrate the impact of TRPC3 on LV compliance and flexibility and, focusing on the TRPC3-Nox2 complex, provide a strategy for prevention of doxorubicin-induced cardiomyopathy.


Wet-spinning of recombinant silk-elastin-like protein polymer fibers with high tensile strength and high deformability.
A recombinant silk-elastin-like protein copolymer SELP-47K containing tandemly repeated amino acid sequence blocks from silk, GAGAGS, and elastin, GVGVP, was fabricated into microdiameter fibers using a wet-spinning technique. Raman spectral analysis revealed the formation of antiparallel beta-sheet crystals of the silk-like blocks. Dry SELP-47K fibers display the dependence of mechanical properties such as Young's modulus on fiber diameter, suggesting more oriented and crystallized molecular chains in small-diameter fibers. Additionally, a brittle fracture mode was identified for dry fibers by SEM analysis of fracture surfaces. Hydration dramatically influenced the mechanical behavior of SELP-47K fibers. In contrast to the high tensile strength and limited strains to failure of dry fibers, fully hydrated SELP-47K fibers possessed strains to failure as high as 700%. Furthermore, upon chemical cross-linking, a tensile mechanical strength up to 20 MPa was achieved in hydrated fibers without compromising their high deformability. By combing the silk- and elastin-derived sequences into a single SELP-47K protein polymer, we demonstrated that protein fibers with high tensile strength and high deformability can be fabricated.