Contractility of the rabbit abdominal aorta 4 days after endothelium denudation.
The purpose of this study was to evaluate contractility of the rabbit abdominal aorta 4 days after de-endothelialization by balloon catheter. The isometric tension of ring-segments in response to vasoactive agents was monitored. A significant enhancement of contraction to noradrenaline and serotonin was found in aortas 4 days after endothelium denudation as compared with controls with endothelium. The enhancement, however, did not differ from that found already in acutely denuded vessels (immediate denudation). No significant difference in contractility to potassium chloride was found in either group of denuded preparations as compared with controls. The sensitivity to all three vasoactive agents (EC50) was not influenced by denudation. These results indicate that changes in the contractility of denuded vessels are predominantly a consequence of lacking the endothelium as a producer of endothelium-derived relaxing factor. The access of mitogens to the media does not seem to interfere with the magnitude of contraction 4 days after denudation.


[Expression of single-chain Fv antibody for anti-beet necrotic yellow vein virus in Escherichia coli].
The heavy chains variable region gene (VH) of monoclonal antibody against beet necrotic yellow vein virus (BNYVV) was amplified from total DNA extracted from anti-BNYVV hybridoma cells by PCR. Sequencing showed that the VH belongs to mouse subgroup II(A) and contains 360 bp, which code one hundred and twenty amino acids. The VH and VL genes were inserted into a plasmid which contains a linker sequence for constructing scFv gene. The new vector named pTC scFv. The scFv was produced in Escherichia coli and appeared binding activity with BNYVV antigen by ELISA method.


Platelet hyper-reactivity in active inflammatory arthritis is unique to the adenosine diphosphate pathway: a novel finding and potential therapeutic target.
To assess the influence of disease activity on platelet function in patients with inflammatory arthritis (IA). Ninety-six patients with an established diagnosis of IA (RA, PsA, seronegative SpA) were recruited. Patients with a history of cardiovascular disease (CVD), diabetes mellitus or receiving anti-platelet therapy were excluded. Demographic data, traditional CVD risk factors and medication use were recorded. Patients were characterized as active disease (n = 38) or control disease (n = 58) groups, respectively, based on internationally validated measures of disease activity [comprising serological markers (ESR, CRP, fibrinogen), patient measures (visual analogue scale of disease activity), evaluator global assessment and the 28-joint disease activity score]. Platelet function was assessed using a novel assay of platelet reactivity. Platelet aggregation to multiple concentrations of arachidonic acid, collagen, epinephrine, thrombin receptor activating peptide and adenosine diphosphate (ADP) were measured simultaneously using a modification of light transmission aggregometry. The two groups (active vs control) were similar in terms of demographics and CVD risk factors. Anti-TNF-alpha therapy use was higher in the control group (P = 0.004), whereas NSAID use was higher in the active group (P = 0.001). There was a significant difference between the two groups in platelet response to ADP (P < 0.001). Platelet aggregation, in response to submaximal concentrations of ADP, was increased in the active disease group compared with the control group. There was no difference in platelet reactivity between the groups in response to any of the other agonists. Patients with active IA demonstrate enhanced platelet reactivity, unique to the ADP pathway. This potential pro-thrombotic bias may contribute to their increased cardiovascular risk.


Synthesis and characterization of protected oligourethanes as crosslinkers of collagen-based scaffolds.
This paper describes the preparation and characterization of water-soluble urethane oligomers bearing protected isocyanate groups. It also points out its ability to crosslink decellularized pericardium, as a model collagen scaffold, and to adjust their structural characteristics. A library of oligourethanes was synthesized by varying the molecular weight (Mw 400, 600, 1000 or 2000 g mol-1) of the poly(ethylene glycol) and the type of aliphatic diisocyanate (isophorone diisocyanate/IPDI or hexamethylene diisocyanate/HDI). 1H and 13C NMR, FTIR and mass spectrometry demonstrated that the crosslinkers are composed of chains with carbamoylsulfonate end groups that have trimeric and pentameric oligourethanes, and monomeric diisocyanate. The degree of crosslinking and hence the in vitro degradation susceptibility of the decellularized pericardium were inversely related to the Mw of the oligourethanes. The toxicity of the extractable products from oligourethane-collagen materials toward fibroblasts and macrophages was found to be lower for the crosslinker derived from IPDI than for those derived from HDI. On the other hand, the resistance to collagenase or oxidative degradation of the bovine pericardium crosslinked with HDI/oligourethane was higher than the one prepared with IPDI/oligourethane. As the Mw of the oligomers regulates the degree of crosslinking while the chemical composition influences the cytocompatibility and biodegradation of decellularized pericardium, these urethane oligomers can be used as safer crosslinkers for other protein-based biomaterials.


Cement dust exposure and acute lung function: a cross shift study.
Few studies have been carried out on acute effects of cement dust exposure. This study is conducted to investigate the associations between current "total" dust exposure and acute respiratory symptoms and respiratory function among cement factory workers. A combined cross-sectional and cross-shift study was conducted in Dire Dawa cement factory in Ethiopia. 40 exposed production workers from the crusher and packing sections and 20 controls from the guards were included. Personal "total" dust was measured in the workers' breathing zone and peak expiratory flow (PEF) was measured for all selected workers before and after the shift. When the day shift ended, the acute respiratory symptoms experienced were scored and recorded on a five-point Likert scale using a modified respiratory symptom score questionnaire. The highest geometric mean dust exposure was found in the crusher section (38.6 mg/m3) followed by the packing section (18.5 mg/m3) and the guards (0.4 mg/m3). The highest prevalence of respiratory symptoms for the high exposed workers was stuffy nose (85%) followed by shortness of breath (47%) and "sneezing" (45%). PEF decreased significantly across the shift in the high exposed group. Multiple linear regression showed a significant negative association between the percentage cross-shift change in PEF and total dust exposure. The number of years of work in high-exposure sections and current smoking were also associated with cross-shift decrease in PEF. Total cement dust exposure was related to acute respiratory symptoms and acute ventilatory effects. Implementing measures to control dust and providing adequate personal respiratory protective equipment for the production workers are highly recommended.


[Control of human African trypanosomiasis in Luba in equatorial Guinea:evaluation of three methods].
The object of this study was to (a) reduce the prevalence of sleeping sickness by serological testing, parasitological examination, and treatment of every infected person; (b) determine the maximum acceptable interval between serological surveys; and (c) define the impact of vector control, using monopyramidal non-impregnated traps, on the transmission. For this sero-parasitological survey, the focus in Luba was divided into three zones as follows: Epicentre A (with high prevalence, 27.5%), Epicentre B (with average prevalence, 8.3%), and Peripheral C (with moderate prevalence, 3.0%). Differences in the prevalence rates in the Epicentres and Peripheral zone permitted the use of three different approaches for control and epidemiological follow-up of the disease: (1) Serological examination of the entire population was carried out by the indirect immunofluorescent antibody test (IFAT), with six-month intervals in Epicentres A and B and once a year in the Peripheral zone C. (2) DIAGNOSIS AND TREATMENT: all IFAT seropositives were examined in Luba hospital for parasites, and every parasitologically confirmed patient was treated according to the WHO protocol of 1983. Another serological test (CATT) was applied to cases in which trypanosomes were not present and if this was positive, the CSF was examined. Cases with parasites and abnormal CSF were treated with melarsoprol, and those with a normal CSF received pentamidine. CATT-negative and parasite-negative cases were considered to be false-positives by IFAT and free of the disease. (3) Vector control: 74 monopyramidal traps (18 traps per km2) were set up in Epicentre A. The flies captured were collected once a month and sent to the programme's laboratory where they were identified and counted.(ABSTRACT TRUNCATED AT 250 WORDS)


Homer expression in the Xenopus tadpole nervous system.
Homer proteins are integral components of the postsynaptic density and are thought to function in synaptogenesis and plasticity. In addition, overexpression of Homer in the developing Xenopus retinotectal system results in axonal pathfinding errors. Here we report that Xenopus contains the homer1 gene, expressed as the isoform, xhomer1b, which is highly homologous to the mammalian homer1b. The mammalian homer1 gene is expressed as three isoforms, the truncated or short form homer1a and the long forms homer1b and -1c. For Xenopus, we cloned three very similar variants of homer1b, identified as Xenopus xhomer1b.1, xhomer1b.2, and xhomer1b.3, which display up to 98% homology with each other and 90% similarity to mammalian homer1b. Furthermore, we demonstrate that Xenopus also contains a truncated form of the Homer1 protein, which could be induced by kainic acid injection and is likely homologous to the mammalian Homer1a. xHomer1b expression was unaffected by neuronal activity levels but was developmentally regulated. Within the brain, the spatial and temporal distributions of both Homer isoforms were similar in the neuropil and cell body regions. Homer1 was detected in motor axons. Differential distribution of the two isoforms was apparent: Homer1b immunoreactivity was prominent at junctions between soma and the ventricular surface; in the retina, the Mueller radial glia were immunoreactive for Homer1, but not Homer1b, suggesting the retinal glia contain only the Homer1a isoform. Homer1b expression in muscle was prominent throughout development and was aligned with the actin striations in skeletal muscle. The high level of conservation of the xhomer1 gene and the protein expression in the developing nervous system suggest that Homer1 expression may be important for normal neuronal circuit development.


Local control of the immune response in the liver.
The physiological function of the liver--such as removal of pathogens and antigens from the blood, protein synthesis and metabolism--requires an immune response that is adapted to these tasks and is locally regulated. Pathogenic microorganisms must be efficiently eliminated while the large number of antigens derived from the gastrointestinal tract must be tolerized. From experimental observations it is evident that the liver favours the induction of tolerance rather than the induction of immunity. The liver probably not only is involved in transplantation tolerance but contributes as well to tolerance to orally ingested antigens (entering the liver with portal-venous blood) and to containment of systemic immune responses (antigen from the systemic circulation entering the liver with arterial blood). This review summarizes the experimental data that shed light on the molecular mechanisms and the cell populations of the liver involved in local immune regulation in the liver. Although hepatocytes constitute the major cell population of the liver, direct interaction of hepatocytes with leukocytes in the blood is unlikely. Sinusoidal endothelial cells, which line the hepatic sinusoids and separate hepatocytes from leukocytes in the sinusoidal lumen, and Kupffer cells, the resident macrophage population of the liver, can directly interact with passenger leukocytes. In the liver, clearance of antigen from the blood occurs mainly by sinusoidal endothelial cells through very efficient receptor-mediated endocytosis. Liver sinusoidal endothelial cells constitutively express all molecules necessary for antigen presentation (CD54, CD80, CD86, MHC class I and class II and CD40) and can function as antigen-presenting cells for CD4+ and CD8+ T cells. Thus, these cells probably contribute to hepatic immune surveillance by activation of effector T cells. Antigen-specific T-cell activation is influenced by the local microenvironment. This microenvironment is characterized by the physiological presence of bacterial constituents such as endotoxin and by the local release of immunosuppressive mediators such as interleukin-10, prostaglandin E2 and transforming growth factor-beta. Different hepatic cell populations may contribute in different ways to tolerance induction in the liver. In vitro experiments revealed that naive T cells are activated by resident sinusoidal endothelial cells but do not differentiate into effector T cells. These T cells show a cytokine profile and a functional phenotype that is compatible with the induction of tolerance. Besides sinusoidal endothelial cells, other cell populations of the liver, such as dendritic cells, Kupffer cells and perhaps also hepatocytes, may contribute to tolerance induction by deletion of T cells through induction of apoptosis.


The needle length of bacterial injectisomes is determined by a molecular ruler.
Size determination represents a fundamental requirement for multicomponent biological structures. Some pathogenic bacteria possess a weapon derived from the flagellum. Like the flagellum, this type-III secretion apparatus, called the injectisome, has a transmembrane basal body, but the external component is a needle-like structure instead of a hook and a filament. Here, we provide evidence that the length of this needle is determined by the size of a protein, YscP, acting as a molecular ruler.


Corpus callosum microstructural organization mediates the effects of physical neglect on social cognition in schizophrenia.
Exposure to early life adversity is associated with both increased risk of developing schizophrenia and poorer performance on measures of social cognitive functioning. In this study, we examined whether interleukin-6 (IL-6) and Corpus Callosum (CC) microstructure mediated the association between childhood physical neglect and social cognition. Fifty-eight patients with a diagnosis of schizophrenia were included. The CANTAB emotion recognition task (unbiased hit rate) was used to assess social cognition. We found that the microstructural organization of the CC significantly mediated the association between physical neglect and emotion recognition. Furthermore, in a sequential mediation analysis that also considered the role of inflammatory response, the association between physical neglect, and lower emotion recognition performance was sequentially mediated by higher IL-6 and lower fractional anisotropy of the CC. This mediating effect of IL-6 was only present when simultaneously considering the effects of CC microstructural organization and remained significant while controlling for the effects of sex, BMI and medication dosage (but not age). Overall, the findings suggest that the association between physical neglect and poorer emotion recognition in schizophrenia occurs, at least in part, via its association with white matter microstructure.


Malonylation is a key reaction in the metabolism of xenobiotic phenolic glucosides in Arabidopsis and tobacco.
Tobacco cells (Nicotiana tabacum L.) accumulate harmful naphthols in the form of malonylated glucosides (Taguchi et al., 2005). Here, we showed that the malonylation of glucosides is a system to metabolize xenobiotics and is common to higher plants. Moreover, some plantlets including Arabidopsis thaliana excreted some of the incorporated naphthols into the culture media as their glucosides. In order to analyze the function of malonylation in the metabolism of these xenobiotics, we identified a malonyltransferase gene (At5g39050) responsible for the malonylation of these compounds in A. thaliana. The recombinant enzyme had malonyltransferase activity toward several phenolic glucosides including naphthol glucosides. A knockout mutant of At5g39050 (pmat1) exposed to naphthols accumulated only a few malonylglucosides in the cell, and released larger amounts of simple glucosides into the culture medium. In contrast, forced expression of At5g39050 in the pmat1 mutant resulted in increased malonylglucoside accumulation and decreased glucoside excretion to the media. The results provided clear evidence of whether the release of glucosides or the storage of malonylglucosides was determined by the At5g39050 expression level. A similar event in naphthol metabolism was observed in the tobacco mutant with a suppressed malonyltransferase gene (NtMaT1). These results suggested that malonylation could be a key reaction to separate the way of xenobiotics disposition, that is, release from cell surface or storage in vacuoles.


Renin-angiotensin system in hypophysectomized rats. I. Control of blood pressure.
The mechanisms whereby the pituitary gland maintains arterial pressure were investigated in rats. The arterial pressure in hypophysectomized rats was 30 mmHg below normal. Saralasin or captopril caused a further fall of 25 and 30 mmHg, respectively, suggesting that the renin-angiotensin system plays a role in blood pressure maintenance in hypophysectomized rats. Growth hormone administration to hypophysectomized rats increased the arterial pressure, but pretreatment with captopril prevented the effect. Plasma renin activity and basal renin secretion (in vitro) was normal in hypophysectomized rats despite a twofold greater renal renin content. Secretory responsiveness to isoproterenol and calcium omission was lower in hypophysectomized rats. It is concluded that the renin-angiotensin system plays a role in maintaining arterial blood pressure in hypophysectomized rats although the responsiveness of the system may be decreased.


Temperature modulates dengue virus epidemic growth rates through its effects on reproduction numbers and generation intervals.
Epidemic growth rate, r, provides a more complete description of the potential for epidemics than the more commonly studied basic reproduction number, R0, yet the former has never been described as a function of temperature for dengue virus or other pathogens with temperature-sensitive transmission. The need to understand the drivers of epidemics of these pathogens is acute, with arthropod-borne virus epidemics becoming increasingly problematic. We addressed this need by developing temperature-dependent descriptions of the two components of r-R0 and the generation interval-to obtain a temperature-dependent description of r. Our results show that the generation interval is highly sensitive to temperature, decreasing twofold between 25 and 35°C and suggesting that dengue virus epidemics may accelerate as temperatures increase, not only because of more infections per generation but also because of faster generations. Under the empirical temperature relationships that we considered, we found that r peaked at a temperature threshold that was robust to uncertainty in model parameters that do not depend on temperature. Although the precise value of this temperature threshold could be refined following future studies of empirical temperature relationships, the framework we present for identifying such temperature thresholds offers a new way to classify regions in which dengue virus epidemic intensity could either increase or decrease under future climate change.


Effects of self-paced high-intensity interval training and moderate-intensity continuous training on the physical performance and psychophysiological responses in recreationally active young adults.
This study aimed to compare the effects of 8-week self-paced high-intensity interval training (HIIT) vs. self-paced moderate-intensity continuous training (MICT) on the physical performance and psychophysiological responses of young adults. Twenty-eight recreationally active young adults (age: 21.1 ± 1.6 years) were randomly assigned to either the self-paced HIIT (n = 14) or the MICT (n = 14) group training protocol. The HIIT consisted of two 12-24 x 30 seconds of high-intensity runs interspersed by 30 seconds of recovery. The MICT completed 24-48 minutes of continuous running. Before and after the 8-week interventions the following tests were completed: maximum oxygen consumption (V̇O2max) estimated from the Yo-Yo Intermittent Recovery Test level 1 (YYIRTL-1), repeated sprint ability (RSA), 10-30-m sprint test, change of direction test (T-drill), countermovement jump (CMJ) and squat jump (SJ), and triple hop distance test (THD). Training rating of perceived exertion (RPE) and physical activity enjoyment scale (PACES) were assessed during the training programme. The HIIT resulted in greater improvement in YYIRTL-1, V̇O2max, RSA and T-drill performances compared to the MICT. Furthermore, RPE and PACES values were higher in the HIIT than the MICT. This study suggested that self-paced HIIT may be a more effective training regime to improve aerobic fitness with greater physical enjoyment in recreationally active young adults.


Pyranose ring conformations in mono- and oligosaccharides: a combined MD and DFT approach.
Among the descriptors of the molecular structure of carbohydrates, the conformation of the pyranose ring is usually the most problematic one to tackle. We present the results of a systematic study oriented at determining the ring-inversion properties of all d-hexopyranoses in the form of monosaccharides, O1-methylated monosaccharides and homotrisaccharides. Contrary to the existing studies, based either on molecular mechanics force fields or on conformational search within ab initio potentials, we combine the structural information from molecular dynamics simulations performed within the GROMOS 56a6CARBO_R force field and use it in a subsequent geometry optimization procedure, performed at the DFT level of theory. This two-step procedure allows avoiding errors resulting from overestimating the contribution of the hydrogen bond-rich, low-energy structures that are not abundant in aqueous solutions. The calculated anomeric ratios and the populations of staggered conformers of the hydroxymethyl group are in satisfactory agreement with the experimental data. Regarding the ring-inversion properties, for the first time, we achieved good agreement of the ab initio-derived data for all hexopyranoses with the experimentally inferred Angyal scheme and with the NMR-inferred populations of ring conformers. The same computational methodology allows determination of the influence of functionalization (methylation or glycosylation) on the ring-inversion properties which includes the influence of the anomeric effect, enhanced upon O1-functionalization. In general, the correlation between ring-inversion properties of unfunctionalized monomers and those of O1-methylated, O1-glycosylated, O4-glycosylated and O1,O4-diglycosylated monomers is qualitatively (but not quantitatively) compatible with that predicted by the classical force fields.


RAB26 coordinates lysosome traffic and mitochondrial localization.
As they mature, professional secretory cells like pancreatic acinar and gastric chief cells induce the transcription factor MIST1 (also known as BHLHA15) to substantially scale up production of large secretory granules in a process that involves expansion of apical cytoplasm and redistribution of lysosomes and mitochondria. How a scaling factor like MIST1 rearranges cellular architecture simply by regulating expression levels of its transcriptional targets is unknown. RAB26 is a MIST1 target whose role in MIST1-mediated secretory cell maturation is also unknown. Here, we confirm that RAB26 expression, unlike most Rabs which are ubiquitously expressed, is tissue specific and largely confined to MIST1-expressing secretory tissues. Surprisingly, functional studies showed that RAB26 predominantly associated with LAMP1/cathepsin D lysosomes and not directly with secretory granules. Moreover, increasing RAB26 expression - by inducing differentiation of zymogen-secreting cells or by direct transfection - caused lysosomes to coalesce in a central, perinuclear region. Lysosome clustering in turn caused redistribution of mitochondria into distinct subcellular neighborhoods. The data elucidate a novel function for RAB26 and suggest a mechanism for how cells could increase transcription of key effectors to reorganize subcellular compartments during differentiation.


Tumor necrosis factor alpha blockade exacerbates murine psoriasis-like disease by enhancing Th17 function and decreasing expansion of Treg cells.
Patients with psoriasis and psoriatic arthritis respond well to tumor necrosis factor alpha (TNFalpha) blockers in general; however, there is now mounting evidence that a small cohort of patients with rheumatoid arthritis who receive TNFalpha blockers develop psoriasis. This study was undertaken to explore the mechanisms underlying TNFalpha blockade-induced exacerbation of skin inflammation in murine psoriasis-like skin disease. Skin inflammation was induced in BALB/c scid/scid mice after they received CD4+CD45RB(high)CD25- (naive CD4) T cells from donor mice. These mice were treated with either anti-interleukin-12 (anti-IL-12)/23p40 antibody or murine TNFRII-Fc fusion protein and were examined for signs of disease, including histologic features, various cytokine levels in the serum, and cytokine or FoxP3 transcripts in the affected skin and draining lymph node (LN) cells. In a separate study, naive CD4+ T cells were differentiated into Th1 or Th17 lineages with anti-CD3/28 magnetic beads and appropriate cytokines in the presence or absence of TNFalpha. Cytokine gene expression from these differentiated cells was also determined. Neutralization of TNFalpha exacerbated skin inflammation and markedly enhanced the expression of the proinflammatory cytokines IL-1beta, IL-6, IL-17, IL-21, and IL-22 but suppressed FoxP3 expression in the skin and reduced the number of FoxP3-positive Treg cells in the draining LNs. TNFalpha also demonstrated a divergent role during priming and reactivation of naive T cells. These results reveal a novel immunoregulatory role of TNFalpha on Th17 and Treg cells in some individuals, which may account for the exacerbation of skin inflammation in some patients who receive anti-TNF treatments.


Growth cycle-dependent pharmacodynamics of antichlamydial drugs.
Chlamydiae are obligate intracellular pathogens that exhibit an extensive intracellular developmental cycle in vivo. Clinical treatment of chlamydial infection is typically initiated upon occurrence of symptomatology and is directed against an asynchronous population of different chlamydial developmental forms. Pharmacodynamics of antichlamydial drugs are predominantly characterized by MICs; however, in vitro determinations of MIC may not reflect differential susceptibilities of the developmental cycle. In this study, we correlated the antichlamydial effect of erythromycin, rifampin, doxycycline, and ciprofloxacin with the developmental stage of a fast-replicating and a slow-replicating chlamydial species. In addition, we describe the influence of concentration on killing. Extracellular elementary bodies and very-early-phase and late-phase chlamydiae were refractory to all tested antibiotics except rifampin, which was very effective against early-cycle chlamydiae. Rifampin was the most effective antibiotic overall, killed in a dose dependent matter, and exhibited moderate synergism with erythromycin. These considerations provide important information on chlamydial biology and antimicrobial susceptibility. A combinational therapy of rifampin and a macrolide should be considered in therapy-refractory infections.


Stabilization of 4E-BP1 by PI3K kinase and its involvement in CHK2 phosphorylation in the cellular response to radiation.
Objectives: 4E-BP1 is a family member of eIF4E binding proteins (4E-BPs) which act as the suppressors of cap-dependent translation of RNA via competitively associating with cap-bound eIF4E. RNA translation regulation is an important manner to control the cellular responses to a series of stress conditions such as ionizing radiation (IR)-induced DNA damage response and cell cycle controlling. This study aimed to determine the mechanism of 4E-BP1 stabilization and its potential downstream target(s) in the response to IR. Methods: PI3Ks kinase inhibitors were used to determine the signaling control of 4E-BP1 phosphorylation and protein stability. shRNA strategy was employed to silence the expression of 4E-BP1 in HeLa and HepG2 cells, and determine its effect on the irradiation-induced CHK2 phosphorylation. The protein degradation/stability was investigated by western blotting on the condition of blocking novel protein synthesis by cycloheximide (CHX). Results: The phosphorylation of 4E-BP1 at Thr37/46 was significantly increased in both HepG2 and HeLa cells by ionizing radiation. Depression of 4E-BP1 by shRNA strategy resulted in an incomplete G2 arrest at the early stage of 2 hours post-irradiation, as well as a higher accumulation of mitotic cells at 10 and 12 hours post-irradiation as compared to the control cells. Consistently, the CHK2 phosphorylation at Thr68 induced by IR was also attenuated by silencing 4E-BP1 expression. Both PI3K and DNA-PKcs kinase inhibitors significantly decreased the protein level of 4E-BP1, which was associated with the accelerated degradation mediated by ubiquitination-proteasome pathway. Conclusion: PI3K kinase activity is necessary for maintaining 4E-BP1 stability. Our results also suggest 4E-BP1 a novel biological role of regulating cell cycle G2 checkpoint in responding to IR stress in association with controlling CHK2 phosphorylation.


The genetics of NOD-like receptors in Crohn's disease.
The first Crohn's disease (CD) susceptibility gene identified was CARD15, which is a member of the emerging NOD-like receptor (NLR) family. These function as intracellular cystosolic pattern recognition receptors (PRRs) and play a central role in the innate immune response. We studied other members of the NLR family using a gene-wide haplotype tagging approach in a well-characterised collection of 547 CD patients and 465 controls. Four single nucleotide polymorphisms (SNPs) in NLRP3 had P values < 0.05 and are in high linkage disequilibrium (LD) with each other (r(2) > 0.90 for all four SNPs). rs4925648 and rs10925019 were the most strongly associated with CD susceptibility (P = 0.001, odds ratio (OR) 1.62, 95% CI 1.2-2.18; and P = 6.5 x 10(-4), OR 1.65, 95% CI 1.23-2.19, respectively). rs1363758 located in NLRP11 was associated with CD susceptibility [P = 0.002 (1.64, 1.19-2.25)], which was weakly confirmed in an independent case-cohort collection on joint analysis [P = 0.05, (1.28, 1-1.64)]. On sub-phenotype analysis, an interesting association between NLRP1 and skin extra-intestinal manifestations and colonic, inflammatory CD was identified. None of these results was replicated in the Wellcome Trust Case Control Consortium study and therefore need replication in a further large cohort.


Drosophila Anillin is unequally required during asymmetric cell divisions of the PNS.
During Drosophila embryogenesis, timely and orderly asymmetric cell divisions ensure the correct number of each cell type that make up the sensory organs of the larval PNS. We report a role of scraps, Drosophila Anillin, during these divisions. Anillin, a constitutive member of the contractile ring is essential for cytokinesis in Drosophila and vertebrates. During embryogenesis we find that zygotically transcribed scraps is required specifically for the unequal cell divisions, those in which cytokinesis occurs in an "off-centred" manner, of the pIIb and pIIIb neuronal precursor cells, but not the equal cell divisions of the lineage related precursor cells. Complementation and genetic rescue studies demonstrate this effect results from zygotic scraps and leads to polyploidy, ectopic mitosis, and loss of the neuronal precursor daughter cells. The net result of which is the formation of incomplete sense organs and embryonic lethality.


Impacts on employment, finances, and lifestyle for working age people facing an expected premature death: A systematic review.
ABSTRACTObjective:The working ages (25-65 years) are a period when most people have significant work, financial, and family responsibilities. A small proportion of working age people will face an expected premature death from cancer or other life-limiting illness. Understanding the impact an expected premature death has on this population is important for informing support. The current study set out to summarize research describing the effects that facing an expected premature death has on employment, financial, and lifestyle of working age people and their families. A systematic review using narrative synthesis approach. Four electronic databases were searched in July 2016 for peer-reviewed, English language studies focusing on the financial, employment, and lifestyle concerns of working age adults living with an advanced life-limiting illness and/or their carers and/or children. Fifteen quantitative and 12 qualitative studies were included. Two-thirds (n = 18) were focused on cancer. All studies identified adverse effects on workforce participation, finances, and lifestyle. Many patients were forced to work less or give up work/retire early because of symptoms and reduced functioning. In addition to treatment costs, patients and families were also faced with child care, travel, and home/car modification costs. Being younger was associated with greater employment and financial burden, whereas having children was associated with lower functional well-being. Changes in family roles were identified as challenging regardless of diagnosis, whereas maintaining normalcy and creating stability was seen as a priority by parents with advanced cancer. This review is limited by the smaller number of studies focussing on the needs of working age people with nonmalignant disease. Working age people facing an expected premature death and their families have significant unmet financial, employment, and lifestyle needs. Comparing and contrasting their severity, timing, and priority for people with nonmalignant conditions is required to better understand their unique needs.


Ecotoxicological impact of dinotefuran insecticide and its metabolites on non-targets in agroecosystem: Harnessing nanotechnology- and bio-based management strategies to reduce its impact on non-target ecosystems.
The class of insecticides known as neonicotinoid insecticides has gained extensive application worldwide. Two characteristics of neonicotinoid pesticides are excellent insecticidal activity and a wide insecticidal spectrum for problematic insects. Neonicotinoid pesticides can also successfully manage pest insects that have developed resistance to other insecticide classes. Due to its powerful insecticidal properties and rapid plant absorption and translocation, dinotefuran, the most recent generation of neonicotinoid insecticides, has been widely used against biting and sucking insects. Dinotefuran has a wide range of potential applications and is often used globally. However, there is growing evidence that they negatively impact the biodiversity of organisms in agricultural settings as well as non-target organisms. The objective of this review is to present an updated summary of current understanding regarding the non-target effects of dinotefuran; we also enumerated nano- and bio-based mitigation and management strategies to reduce the impact of dinotefuran on non-target organisms and to pinpoint knowledge gaps. Finally, future study directions are suggested based on the limitations of the existing studies, with the goal of providing a scientific basis for risk assessment and the prudent use of these insecticides.


Rapid determination of Epstein-Barr virus-specific CD8(+) T-cell frequencies by flow cytometry.
We have developed an efficient and rapid method for detection of Epstein-Barr virus (EBV)-specific CD8(+) T-cell frequencies both in freshly isolated peripheral blood mononuclear cells (PBMCs) and in vitro established cytotoxic T lymphocyte (CTL) lines. Responder cells are thereby stimulated with an autologous lymphoblastoid cell line for 5 hours and intracellular accumulation of interferon gamma (IFNgamma) is detected by multiparameter flow cytometric analysis. EBV-specific CD8(+) T-cell frequencies ranged between 0.63% and 1. 29% in PBMCs of 5 healthy long-term EBV carriers. Using EBV-specific T-cell lines, it was shown that flow cytometric analysis is more sensitive than limiting dilution analysis for CTL precursors and enzyme-linked immunospot assay detecting IFNgamma-producing T cells. The class I restriction of IFNgamma production was confirmed using an anti-class I monoclonal antibody (MoAb). Information on other cytokine production of EBV-specific CTLs could be obtained using combinations of anti-cytokine MoAbs. The sensitive and rapid nature of the flow cytometric assay for EBV-specific CD8(+) T-cell frequency has significant advantages for evaluation of EBV-specific CD8(+) T-cell responses in PBMCs of patients with EBV-related diseases.


The occipitoparietal pathway of the macaque monkey: comparison of pyramidal cell morphology in layer III of functionally related cortical visual areas.
The dendritic morphology of pyramidal cells located at the base of layer III in the primary visual area (V1), the second visual area (V2), the middle temporal area (MT), the ventral portion of the lateral intraparietal area (LIPv) and in the portion of cytoarchitectonic area 7a within the anterior bank of the superior temporal sulcus was revealed by injecting neurons with Lucifer Yellow in fixed, flattened slices of macaque monkey visual cortex. These areas correspond to different levels of the occipitoparietal cortical 'stream', which processes information related to motion and spatial relationships in the visual field. The tissue was immunocytochemically processed to obtain a light-stable diaminobenzidine reaction product, revealing the dendritic morphology in fine detail. Retrogradely labelled MT-projecting neurons in supragranular V1 (layer IIIc of Hassler's nomenclature, corresponding to Brodmann's layer IVb) were predominantly pyramidal, although many spiny multipolar (stellate) cells were also found. The average basal dendritic field area of pyramidal neurons in sublamina IIIc of V1 was significantly smaller than that in the homologous layer of V2, within the cytochrome oxidase-rich thick stripes. Furthermore, the average basal dendritic field areas of V1 and V2 pyramidal neurons were significantly smaller than those of neurons in MT, LIPv and area 7a. There was no difference in basal dendritic field area between layer III pyramidal neurons in areas MT, LIPv and 7a. While the shape of most basal dendritic fields was circularly symmetrical in the dimension tangential to the cortical layers, there were significant biases in complexity, with dendritic branches tending to cluster along particular axes. Sholl analysis revealed that the dendritic fields of neurons in areas MT, LIPv and 7a were significantly more complex (i.e. had a larger number of branches) than those of V1 or V2 neurons. Analysis of basal dendritic spine densities revealed regional variations along the dendrites, with peak densities being observed 40-130 microns from the cell body, depending on the visual area. The peak spine density of layer III pyramidal neurons in V1 was lower than that observed in V2, MT or LIPv, which were all similar. Pyramidal neurons in area 7a had the greatest peak spine density, which was on average 1.7 times that found in V1. Calculations based on the average spine density and number of dendritic branches at different distances from the cell body demonstrated a serial increase in the total number of basal dendritic spines per neuron at successive stations of the occipitoparietal pathway. Our observations, comparing dendritic fields of neurons in the homologous cortical layer at different levels of a physiologically defined 'stream', indicate changes in pyramidal cell morphology between functionally related areas. The relatively large, complex, spine-dense dendritic fields of layer III pyramidal cells in rostral areas of the occipitoparietal pathway allow these cells to sample a greater number of more diverse inputs in comparison with cells in 'lower' areas of the proposed hierarchy.


Impact of obstructive sleep apnoea on diastolic function.
We investigated whether obstructive sleep apnoea (OSA) independently affects diastolic function in a primary care cohort of patients with cardiovascular risk factors. 378 study participants with risk factors for diastolic dysfunction were prospectively included and a polygraphy was performed in all patients. Diastolic dysfunction was assessed by comprehensive echocardiography including tissue Doppler. Sleep apnoea was classified according to apnoea/hypopnoea index (AHI) as none (AHI <5 events·h(-1)), mild (AHI ≤5 to <15 events·h(-1)) or moderate-to-severe (AHI ≥15 events·h(-1)). Patients with central sleep apnoea (n=14) and patients with previously diagnosed sleep apnoea (n=12) were excluded. In the remaining 352 subjects, 21.6% had an AHI ≥15 events·h(-1). The prevalence of diastolic dysfunction increased with the severity of sleep apnoea from 44.8% (none) to 56.8% (mild) to 69.7% (moderate-to-severe sleep apnoea) (p=0.002). The degree of diastolic dysfunction also increased with sleep apnoea severity (p=0.004). In univariate regression analysis, age, desaturation index, AHI, cardiac frequency, angiotensin receptor 1 antagonist therapy, body mass index (BMI) and left ventricular mass were associated with diastolic dysfunction. In multivariate regression analysis, only age, BMI, AHI and cardiac frequency were independently associated with diastolic dysfunction. Moderate-to-severe OSA is independently associated with diastolic dysfunction in patients with classical risk factors for diastolic dysfunction.


The activation of SRC family kinases and focal adhesion kinase with the loss of the amplified, mutated EGFR gene contributes to the resistance to afatinib, erlotinib and osimertinib in human lung cancer cells.
Second- and third-generation inhibitors of epidermal growth factor receptor (EGFR) tyrosine kinase activity (EGFR-TKIs) are improving the treatment of patients with non-small cell lung cancer. Here we established two sublines (BR1-8 and BR2-3) resistant to a second-generation inhibitor, afatinib, from the human lung cancer cell line HCC827 that harbors a mutation that activates the tyrosine kinase activity of EGFR. These afatinib-resistant sublines were resistant to first-generation EGFR-TKIs, gefitinib and erlotinib, and a third-generation EGFR-TKI, osimertinib. These resistant sublines showed markedly reduced levels of multiple EGFR family proteins, including the activated mutant EGFR, and complete loss of EGFR amplification as compared with their parental HCC827 cells harboring amplification of EGFR gene. Treatment with the multikinase inhibitor dasatinib or transfection with a SRC small interfering RNA inhibited cell survival and AKT phosphorylation in drug-resistant sublines to a greater extent compared with HCC827 cells. Further, the migration of drug-resistant cells was greater compared with that of HCC827 cells and was inhibited by dasatinib or an FAK inhibitor. These findings indicate that compensatory activation of SRC family kinases (SFKs) and FAK supports the survival and migration of afatinib-resistant cells when the expression of multiple EGFR family proteins was mostly abrogated. Combinations of potent drugs that target SFKs and FAK may overcome the resistance of lung cancer cells to second-generation TKIs.


Acupoint selection rules of staging, syndrome types, and symptoms of acupuncture for peripheral facial paralysis based on data mining.
To analyze and explore the acupoint selection rules of different staging, syndrome types, and symptoms of acupuncture for peripheral facial paralysis based on data mining. Literature regarding acupuncture for peripheral facial paralysis was retrieved in CNKI, VIP, Wanfang, SinoMed, and PubMed from January 1, 2012, to December 31, 2021. The Traditional Chinese Medicine Inheritance Assistant Platform (V 3.0), SPSS Modeler 18.0, and Cytoscape software were used to analyze the selection of acupoints based on staging, meridian distribution of acupoints, the selection of acupoints based on syndrome types, syndrome types based on staging, and symptom-based acupoint selection. Association rule analysis was performed for each stage and major syndrome types, and co-occurrence network diagrams were generated. A total of 1 695 articles were included, involving 124 acupoints with a total frequency of 5 456 times. Among 1 080 articles related to staging-based acupoint selection, the acute stage had the highest acupoint use frequency (2 224 times) and number (88 acupoints). Among all stages, the acupoints of the stomach meridian of foot-yangming were most frequently used, with Dicang (ST 4)-Yangbai (GB 14) and Dicang (ST 4)-Jiache (ST 6) being the high-frequency acupoint pairs in each stage. The top four syndrome types in terms of acupoint usage frequency were wind-cold syndrome, wind-heat syndrome, qi-blood deficiency syndrome, and liver-gallbladder damp-heat syndrome. Among 521 articles combining staging with symptoms or syndrome types with symptoms for acupoint selection, the symptom "nasolabial groove becoming shallow or disappearing" had the highest frequency of occurrence. The symptoms with the highest acupoint usage frequency and number were "forehead wrinkles becoming shallow or disappearing" and "pain behind the ear" respectively. Acupoint selection for peripheral facial paralysis primarily focuses on the yang meridians in each stage, supplemented by local acupoints and based on the pathological characteristics. Syndrome differentiation and treatment should be prioritized, combined with local acupoint selection. Clinical acupoint selection can be based on a combination of staging, syndrome types, and symptoms.


Engineering pH-Sensitive Stable Nanovesicles for Delivery of MicroRNA Therapeutics.
MicroRNAs (miRNAs) are small non-coding endogenous RNAs, which are attracting a growing interest as therapeutic molecules due to their central role in major diseases. However, the transformation of these biomolecules into drugs is limited due to their unstability in the bloodstream, caused by nucleases abundantly present in the blood, and poor capacity to enter cells. The conjugation of miRNAs to nanoparticles (NPs) could be an effective strategy for their clinical delivery. Herein, the engineering of non-liposomal lipid nanovesicles, named quatsomes (QS), for the delivery of miRNAs and other small RNAs into the cytosol of tumor cells, triggering a tumor-suppressive response is reported. The engineered pH-sensitive nanovesicles have controlled structure (unilamellar), size (<150 nm) and composition. These nanovesicles are colloidal stable (>24 weeks), and are prepared by a green, GMP compliant, and scalable one-step procedure, which are all unavoidable requirements for the arrival to the clinical practice of NP based miRNA therapeutics. Furthermore, QS protect miRNAs from RNAses and when injected intravenously, deliver them into liver, lung, and neuroblastoma xenografts tumors. These stable nanovesicles with tunable pH sensitiveness constitute an attractive platform for the efficient delivery of miRNAs and other small RNAs with therapeutic activity and their exploitation in the clinics.


Perivenous expression of the mRNA of the three hypoxia-inducible factor alpha-subunits, HIF1alpha, HIF2alpha and HIF3alpha, in rat liver.
The cDNAs of three hypoxia-inducible factor (HIF) alpha-subunits were cloned from RNA of primary rat hepatocytes by reverse transcriptase PCR. All three cDNAs encoded functionally active proteins, of 825, 874 and 662 amino acids. After transfection they were able to activate luciferase activity of a luciferase gene construct containing three HIF-responsive elements. The mRNAs of the rat HIF alpha-subunits were expressed predominantly in the perivenous zone of rat liver tissue; the nuclear HIFalpha proteins, however, did not appear to be zonated.


Barriers to reporting of adverse drugs reactions: a cross sectional study among community pharmacists in United Kingdom.
Adverse Drug Reactions (ADRs) are a major public health problem. Prompt reporting of suspected ADRs is fundamental in the post-marketing surveillance of medicines and helps in ensuring medicine safety. However, fewer ADRs are reported in general and in particular by community pharmacists. There is limited knowledge about the factors which are preventing community pharmacists in the UK from reporting an ADR. To identify the barriers to ADR reporting among community pharmacists practicing in the UK. A cross sectional study using a 25-items questionnaire (both online and paper based) including 10 barriers to ADR reporting was conducted from 1st April 2012 to September 2012. Community pharmacists practicing in the West Midlands, UK, were approached for the participation in this study. Chi-Square and regression were applied to identify covariates for the barriers to ADR reporting. A significant value of 0.05 was assigned for analysis. Of the 230 invited community pharmacists, 138 pharmacists responded (response rate 60%). The median age of respondents was 31 years. All pharmacists reported that they would report both serious and mild ADRs from drugs with black triangle among children as well as adults. About 95% (n=131) of the pharmacists were familiar with the paper based ADR reporting system. Store-based pharmacists were more likely to be more confident about which ADRs to report [0.680, 95% Confidence Interval 0.43-3.59]. Lack of time 46.4% (n=64), and pharmacists perception that ADR is not serious enough to report (65.2%; n=90) were identified as barriers to ADR reporting. Majority 63.0% (n=87) of the pharmacists identified training and information about what to report and access to Information Technology (IT) (For example access to internet connection) 61.6% (n=85) as facilitators to ADR reporting process. Lack of time and ADRs considered not serious enough by pharmacists to report were barriers to ADR reporting. Further training and education about the types of ADRs to be reported can help to improve the reporting of ADRs.


Altered (neo-) lacto series glycolipid biosynthesis impairs α2-6 sialylation on N-glycoproteins in ovarian cancer cells.
The (neo-) lacto series glycosphingolipids (nsGSLs) comprise of glycan epitopes that are present as blood group antigens, act as primary receptors for human pathogens and are also increasingly associated with malignant diseases. Beta-1, 3-N-acetyl-glucosaminyl-transferase 5 (B3GNT5) is suggested as the key glycosyltransferase for the biosynthesis of nsGSLs. In this study, we investigated the impact of CRISPR-Cas9 -mediated gene disruption of B3GNT5 (∆B3GNT5) on the expression of glycosphingolipids and N-glycoproteins by utilizing immunostaining and glycomics-based PGC-UHPLC-ESI-QTOF-MS/MS profiling. ∆B3GNT5 cells lost nsGSL expression coinciding with reduction of α2-6 sialylation on N-glycoproteins. In contrast, disruption of B4GALNT1, a glycosyltransferase for ganglio series GSLs did not affect α2-6 sialylation on N-glycoproteins. We further profiled all known α2-6 sialyltransferase-encoding genes and showed that the loss of α2-6 sialylation is due to silencing of ST6GAL1 expression in ∆B3GNT5 cells. These results demonstrate that nsGSLs are part of a complex network affecting N-glycosylation in ovarian cancer cells.


[Ovarian vein syndrome. Report of 2 cases].
To report the clinicae characteristics and treatment of the ovarian vein syndrome. We studied two cases of female patients with obstructive uropathy secondary to ovarian vein syndrome cared for at our department over the last 4 years. Both cases were treated surgically. Postoperative outcomes were satisfactory. The ovarian vein syndrome is rare. Surgery is the treatment of choice for symptomatic cases.


HIV-1-Specific Immunodominant T-Cell Responses Drive the Dynamics of HIV-1 Recombination Following Superinfection.
A series of HIV-1 CRF01_AE/CRF07_BC recombinants were previously found to have emerged gradually in a superinfected patient (patient LNA819). However, the extent to which T-cell responses influenced the development of these recombinants after superinfection is unclear. In this study, we undertook a recombination structure analysis of the gag, pol, and nef genes from longitudinal samples of patient LNA819. A total of 9 pol and 5 nef CRF01_AE/CRF07_BC recombinants were detected. The quasispecies makeup and the composition of the pol and nef gene recombinants changed continuously, suggestive of continuous evolution in vivo. T-cell responses targeting peptides of the primary strain and the recombination regions were screened. The results showed that Pol-LY10, Pol-RY9, and Nef-GL9 were the immunodominant epitopes. Pol-LY10 overlapped with the recombination breakpoints in multiple recombinants. For the LY10 epitope, escape from T-cell responses was mediated by both recombination with a CRF07_BC insertion carrying the T467E/T472V variants and T467N/T472V mutations originating in the CRF01_AE strain. In pol recombinants R8 and R9, the recombination breakpoints were located ~23 amino acids upstream of the RY9 epitope. The appearance of new recombination breakpoints harboring a CRF07_BC insertion carrying a R984K variant was associated with escape from RY9-specific T-cell responses. Although the Nef-GL9 epitope was located either within or 10~11 amino acids downstream of the recombination breakpoints, no variant of this epitope was observed in the nef recombinants. Instead, a F85V mutation originating in the CRF01_AE strain was the main immune escape mechanism. Understanding the cellular immune pressure on recombination is critical for monitoring the new circulating recombinant forms of HIV and designing epitope-based vaccines. Vaccines targeting antigens that are less likely to escape immune pressure by recombination and/or mutation are likely to be of benefit to patients with HIV-1.


TRAM, a predicted RNA-binding domain, common to tRNA uracil methylation and adenine thiolation enzymes.
A previously undetected conserved domain is identified in two distinct classes of tRNA-modifying enzymes, namely uridine methylases of the TRM2 family and enzymes of the MiaB family that are involved in 2-methylthioadenine formation. This domain, for which the acronym TRAM is proposed after TRM2 and MiaB, is predicted to bind tRNA and deliver the RNA-modifying enzymatic domains to their targets. In addition to the two families of RNA-modifying enzymes, the TRAM domain is present in several other proteins associated with the translation machinery and in a family of small, uncharacterized archaeal proteins that are predicted to have a role in the regulation of tRNA modification or translation. Secondary structure prediction indicates that the TRAM domain adopts a simple beta-barrel fold. In addition, sequence analysis of the MiaB family enzymes showed that they share the predicted catalytic site with biotin and lipoate synthases and probably employ the same mechanism for sulfur insertion into their respective substrate.


The endoplasmic reticulum membrane-bending protein RETICULON facilitates chloroplast relocation movement in Marchantia polymorpha.
Plant cells alter the intracellular positions of chloroplasts to ensure efficient photosynthesis, a process controlled by the blue light receptor phototropin. Chloroplasts migrate toward weak light (accumulation response) and move away from excess light (avoidance response). Chloroplasts are encircled by the endoplasmic reticulum (ER), which forms a complex network throughout the cytoplasm. To ensure rapid chloroplast relocation, the ER must alter its structure in conjunction with chloroplast relocation movement, but little is known about the underlying mechanism. Here, we searched for interactors of phototropin in the liverwort Marchantia polymorpha and identified a RETICULON (RTN) family protein; RTN proteins play central roles in ER tubule formation and ER network maintenance by stabilizing the curvature of ER membranes in eukaryotic cells. Marchantia polymorpha RTN1 (MpRTN1) is localized to ER tubules and the rims of ER sheets, which is consistent with the localization of RTNs in other plants and heterotrophs. The Mprtn1 mutant showed an increased ER tubule diameter, pointing to a role for MpRTN1 in ER membrane constriction. Furthermore, Mprtn1 showed a delayed chloroplast avoidance response but a normal chloroplast accumulation response. The live cell imaging of ER dynamics revealed that ER restructuring was impaired in Mprtn1 during the chloroplast avoidance response. These results suggest that during the chloroplast avoidance response, MpRTN1 restructures the ER network and facilitates chloroplast movement via an interaction with phototropin. Our findings provide evidence that plant cells respond to fluctuating environmental conditions by controlling the movements of multiple organelles in a synchronized manner.


Alpha fetoprotein mediates HBx induced carcinogenesis in the hepatocyte cytoplasm.
Although tumor-associated fetal protein AFP has demonstrated utility as a clinical tumor marker, the significance of intracellular AFP is still unclear. The aim of this study was to explore the role of cytoplasmic AFP during HBx induced carcinogenesis, which had not previously been recognized; 614 HCC patients were analyzed for correlation of HBV infection with AFP level, and much higher AFP levels were found in HBsAg positive patients. Tumor tissue specimens from 20 HCC patients were used for analysis of AFP and GADD45α. Analysis of HCC specimens showed that upregulation of cytoplasmic AFP is associated with down-regulation of GADD45α in neoplastic tissue. Transfected HBx promotes transcription of AFP by acting on the elements in the AFP gene regulatory region. HBx itself did not directly impact transcription of GADD45α. However, the obstruction of RAR signaling by HBx induced elevation of AFP, which led to down-regulation of GADD45α. Cytoplasmic AFP was able to interact with RAR, disrupting its entrance into the nucleus and binding to the elements in the regulatory region of the GADD45α gene. Knockdown of AFP in siRNA-transfected AFP positive cell lines was synchronously associated with an incremental increase of RAR binding to DNA, as well as upregulation of GADD45α and it was contrary in AFP gene-transfected AFP negative cell lines. These results indicate cytoplasmic AFP is not only a histochemical tumor biomarker for human hepatoma but is also an intracellular signal molecule and potential participant in HBx induced hepatocarcinogenesis.


The role of ultrasound evaluation in the detection of early-stage epithelial ovarian cancer.
Epithelial ovarian cancer kills more women than all other gynecologic malignancies combined because of our inability to detect early-stage disease. Ultrasonography has demonstrated usefulness in the detection of ovarian cancer in asymptomatic women, but its value for the detection of early-stage epithelial ovarian cancer in women of increased risk is uncertain. We examined the usefulness of sonography in the detection of early-stage epithelial ovarian cancer in asymptomatic high-risk women who participated in the National Ovarian Cancer Early Detection Program. Only asymptomatic women of increased risk for the development of ovarian cancer with initial normal gynecologic and ultrasound examinations were eligible to participate in the institutional review board-approved National Ovarian Cancer Early Detection Program. Participants underwent comprehensive gynecologic and ultrasound examinations every 6 months. Increased risk includes women with at least 1 affected first-degree relative with ovarian cancer; a personal history of breast, ovarian, or colon cancer; > or =1 affected first- and second-degree relatives with breast and or ovarian cancer; inheritance of a breast cancer mutation from an affected family member, or membership within a recognized cancer syndrome. The average age of the 4526 women who were evaluated was 46 years; 2610 women were premenopausal, and 1916 women were postmenopausal. A total of 12,709 scans have been performed since 1990. Visualization of both ovaries was noted in 98% of premenopausal and in 94% of postmenopausal women. Fourteen women had undergone unilateral salpingo-oophorectomy. Recall rates at less than the routine 6-month interval were 0.4% in the premenopausal and 0.3% in postmenopausal women. A total of 98 women with persistent adnexal masses were identified, and 49 invasive surgical procedures were performed that diagnosed 37 benign ovarian tumors and 12 gynecologic malignancies. All cancers were detected in asymptomatic women who had normal ultrasound and physical examinations 12 and 6 months before the cancer diagnosis. The detected malignancies were fallopian tube carcinoma (stage IIIC; n = 4 women), primary peritoneal carcinoma (n = 4 women; stage IIIA, 1 woman; stage IIIB, 2 women; stage IIIC, 1 woman), epithelial ovarian cancer (stages IIIA and IIIB; n = 2 women), and endometrial adenocarcinoma (stage IA; n = 2 women). Additionally 37 primary and 12 recurrent breast carcinomas were detected by physical examination. A total of 184 women with genetic predisposition (breast cancer positive) have undergone a prophylactic bilateral salpingo-oophorectomy; 23% of these procedures found atypical hyperplasia, and unexpectedly, 2 women (1%) were found to have stage III (A and B) primary peritoneal carcinoma. This study demonstrates the limited value of diagnostic ultrasound examination as an independent modality for the detection of early-stage epithelial ovarian cancer in asymptomatic women who are at increased risk for disease.


An observational analysis of profile and outcome of children with malnutrition admitted at malnutrition treatment centre.
Nutritional status is an important health indicator and children bear maximum burden of it. Poor nutrition during first thousand days of a child's life can lead to stunted growth. Comorbid condition adversely affects treatment outcome of malnutrition. This study is aimed to describe the profile and treatment outcome of malnourished children. This record-based retrospective, analytical observational study was conducted from October to December 2020. Data of severe acute malnourished children who were admitted in malnutrition treatment centre during August 2017 to September 2020, collected. Totally, 264 eligible children were enrolled in the study. The data were collected through medical records of patients. Data were entered in excel spreadsheet and analysed. Most of the children belonged to 6-24 months of age, almost equal proportion of male and female, and more than three-fourth of children were from rural area. The mean length of stay in hospital was 15.19 ± 6.64 days and average weight gain of children was 7.6 ± 6.7 gm/kg/day. There was significant increase in Z-score (Weight/Height) and MUAC at the time of discharge (P value <0.05). There was no significant association of treatment outcome with the presence of comorbidities (P value >0.05). Most vulnerable age group for malnutrition was 6 months to 2 years. Majority of children with malnutrition improve with standard treatment protocol of malnutrition. We can improve the ability of community health workers to significantly reduce child malnutrition at the primary care level and in the community by equipping them with the required knowledge, tools, and assistance.


Helicobacter pylori infection: a global occupational risk for healthcare workers?
The occupational risks from Helicobacter pylori, a major cause of gastroduodenal diseases, were reviewed for selected groups of healthcare workers (HCWs). A literature search was performed using Medline/Pubmed, 1983 to June 2003. Additional manual searches were made using reference lists from the selected articles. Current knowledge implies various pathways of agent transmission, favoring person-to-person transmission. The risks are highest for gastroenterologists, some nurses, and employees caring for persons with mental disability. Results for other groups are conflicting.


Cytomorphology of intraductal oncocytic papillary neoplasm of the liver.
We describe the first cytology case report of an intraductal oncocytic papillary neoplasm (IOPN) of the liver. A 51-year-old male presented with recurrent cholangitis. Magnetic resonance imaging and endoscopic retrograde cholangiopancreatogram revealed a 1.1 × 0.9 cm polypoid lesion within the left intrahepatic bile duct. Fine-needle aspiration and needle core biopsy (NCB) revealed nests, 3-dimensional or papillary clusters of columnar or cuboidal cells with loss of polarity. The nuclei were uniform with even chromatin, and cytoplasm was granular or vacuolated. No mitosis or necrosis was seen. The cytologic and histologic diagnosis was "consistent with Intraductal Oncocytic Papillary Neoplasm (IOPN), intermediate grade (borderline)." The patient then underwent a left lateral liver segmentectomy. Microscopic examination showed histology similar to the NCB with no stromal invasion identified. Hepatic IOPN poses a diagnostic challenge due to its broad differential diagnoses. Both malignant and non-malignant IOPNs may present with similar clinical symptoms, pathology, histology, cytomorphology, and immunohistochemistry. Hepatic IOPN should be excised as it is a precursor lesion of adenocarcinoma.


Cerebellar vascular and synaptic responses in normal mice and in transgenics with Purkinje cell dysfunction.
We used transgenic mice with Purkinje cell dysfunction (PO3 line) to study the role of these neurons in the increase in cerebellar blood flow (BFcrb) produced by stimulation of the cerebellar parallel fibers (PF). Mice (age 8-10 wk) were anesthetized (halothane) and artificially ventilated. Arterial pressure and end-tidal CO2 were monitored continuously. Arterial blood gases were measured. The PF were stimulated electrically (100 microA, 30 Hz; 40 s), and the increases in BFcrb were monitored by a laser-Doppler flow probe. First, we characterized the increases in BFcrb and the field potentials produced by PF stimulation in normal mice. PF stimulation evoked the typical field potentials and increased BFcrb by 60 +/- 4% (100 microA, 30 Hz; n = 10). The increases in BFcrb were attenuated by the broad-spectrum glutamate receptor antagonist kynurenate (-84 +/- 3%; P < 0.05 analysis of variance; n = 5), by the DL-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor antagonist 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(f)quinoxaline (-62 +/- 6%; P < 0.05; n = 5), and by the nitric oxide synthase inhibitor N omega-nitro-L-arginine (-46 +/- 7%; P < 0.05; n = 5). In PO3 transgenic mice, the increases in BFcrb produced by PF stimulation were reduced (P < 0.001) at every stimulus intensity and frequency tested (residual increase at 100 microA, 30 Hz: 19 +/- 2%; n = 6). The field potentials evoked by PF stimulation also were abnormal in that they lacked the late negative wave (n = 6), a finding consistent with lack of depolarization of Purkinje cells. The residual flow response in the transgenics was abolished by N omega-nitro-L-arginine (n = 5; P > 0.05). Ultrastructural studies showed that the density of PF-Purkinje cell synapses is reduced in PO3 mice, whereas the morphology of molecular layer interneurons (stellate cells) is normal. The findings suggest that Purkinje cells are responsible for a sizable component of the flow response whereas molecular layer interneurons mediate the remainder of the response. The study provides evidence that mouse mutants with spontaneous or genetically engineered cerebellar abnormalities could be useful to study the cellular and molecular correlates of functional hyperemia in the central nervous system.


Efficacy of low-dose radiotherapy in painful gonarthritis: experiences from a retrospective East German bicenter study.
To evaluate the efficacy of low-dose radiotherapy in painful gonarthritis. We assessed the medical records of 1037 patients with painful gonarthritis who had undergone low-dose radiotherapy between 1981 and 2008. The subjective patient perception of the response to irradiation as graded immediately or up to two months after the completion of a radiotherapy series was evaluated and correlated with age, gender, radiological grading and the duration of symptoms before radiotherapy. Moreover, we performed a mail survey to obtain additional long-term follow-up information and received one hundred and six evaluable questionnaires. We assessed 1659 series of radiotherapy in 1037 patients. In 79.3% of the cases the patients experienced a slight, marked or complete pain relief immediately or up to two months after the completion of radiotherapy. Gender, age and the duration of pain before radiotherapy did not have a significant influence on the response to irradiation. In contrast, severe signs of osteoarthritis were associated with more effective pain relief. In more than 50% of the patients who reported a positive response to irradiation a sustained period of symptomatic improvement was observed. Our results confirm that low-dose radiotherapy is an effective treatment for painful osteoarthritis of the knee. In contrast to an earlier retrospective study, severe signs of osteoarthritis constituted a positive prognostic factor for the response to irradiation. A randomized trial is urgently required to compare radiotherapy with other treatment modalities.


Epidermal growth factor receptor metabolism and protein kinase activity in human A431 cells infected with Snyder-Theilen feline sarcoma virus or harvey or Kirsten murine sarcoma virus.
When human A431 cells, which carry high numbers of epidermal growth factor (EGF) receptors, are exposed to EGF, the total content of phosphotyrosine in cell protein is increased, the EGF receptor becomes phosphorylated at tyrosine, and new phosphotyrosine-containing 36,000- and 81,000-dalton proteins are detected. We examined the properties of A431 cells infected with Snyder-Theilen feline sarcoma virus, whose transforming protein has associated tyrosine protein kinase activity, and Harvey and Kirsten sarcoma viruses, whose transforming proteins do not. In all cases, the infected cells were more rounded and more capable of anchorage-independent growth than the uninfected cells. EGF receptors were assayed functionally by measuring EGF binding and structurally by metabolic labeling and immunoprecipitation. In no case did infection appear to alter the rate of EGF receptor synthesis, but infection reduced EGF receptor stability by about 50% for cloned Harvey sarcoma virus-infected cells and by 80% for cloned feline sarcoma virus-infected cells. The corresponding reductions in EGF binding were 70 and 90%, respectively. The proteins of feline sarcoma virus-infected A431 cells contained an increased amount of phosphotyrosine, and the 36,000- and 81,000-dalton phosphoproteins were detected. The EGF receptor was not detectably phosphorylated at tyrosine, however, unless the cells were exposed to EGF. The Harvey and Kirsten sarcoma virus-infected cells did not exhibit elevated levels of phosphotyrosine either in the total cell proteins or in the EGF receptor, nor were the 36,000- and 81,000-dalton proteins detectable. However, these phosphoproteins were found in the infected cells after EGF treatment. Thus, all of the infected A431 cells exhibited reduced EGF binding and increased degradation of EGF receptors, yet their patterns of protein phosphorylation were distinct from those of EGF-treated A431 cells.


Perioperative glycemic control and postoperative complications in patients undergoing emergency general surgery: What is the role of Plasma Hemoglobin A1c?
Plasma hemoglobin A1c (HbA1c) reflects quality of glucose control in diabetic patients. Literature reports that patients undergoing surgery with an elevated HbA1c level are associated with increased postoperative morbidity and mortality. The aim of our study was to evaluate the impact of HbA1c level on outcomes after emergency general surgery (EGS). We performed a 3-year analysis of our prospectively maintained EGS database. Patients who had HbA1c levels measured within 3 months before surgery were included. Patients were divided into two groups (HbA1c < 6 and HbA1c ≥ 6). Our primary outcome measures included in-hospital complications (major and minor complications), hospital and intensive care unit length of stay, and mortality. Secondary outcomes measures were 30-day complications, readmissions, and mortality. Multivariate and linear regressions were performed. Of the 402 study patients, mean age was 61 ± 12 years, 53% were females, and 63.8% were diabetics. Overall, 49% had an HbA1c ≥ 6%; the mortality rate was 6%. Those with hypertension, history of coronary artery disease, and body mass index of 30 kg/m or greater were more likely to have HbA1c of 6.0% or greater. 7.9% patients experienced major complications. Patients with HbA1c of 6% or greater had a higher complication rate (36% vs 11%, p < 0.001) than those with HbA1c less than 6%. However there was no difference in mortality between two groups (p = 0.09). After controlling for confounders, HbA1c ≥ 6.0% (odds ratio [OR], 2.9; p < 0.01) and a postoperative random blood sugar (RBS) of 200 mg/dL or greater (OR, 2.3; p < 0.01) were independent predictors of major complications. Patients with both HbA1c of 6.0% or greater and postoperative RBS of 200 or greater had higher odds (OR, 4.2; p < 0.01) of developing major complication. After adjusting for confounders, a higher HbA1c was independently correlated with a higher postoperative RBS (b = 0.494, [19.7-28.4], p = 0.02), but there was no correlation with the preoperative RBS. Patients with HbA1c of 6.0% or greater and a postoperative RBS of 200 mg/dL or greater have a four times higher risk of developing major complications after EGS. A preoperative HbA1c can stratify patients prone to develop postoperative hyperglycemia, regardless of their preoperative RBS. Prognostic, level III.


Free-classification of perceptually similar speakers with dysarthria.
In this investigation, the construct of perceptual similarity was explored in the dysarthrias. Specifically, we employed an auditory free-classification task to determine whether listeners could cluster speakers by perceptual similarity, whether the clusters mapped to acoustic metrics, and whether the clusters were constrained by dysarthria subtype diagnosis. Twenty-three listeners blinded to speakers' medical and dysarthria subtype diagnoses participated. The task was to group together (drag and drop) the icons corresponding to 33 speakers with dysarthria on the basis of how similar they sounded. Cluster analysis and multidimensional scaling (MDS) modeled the perceptual dimensions underlying similarity. Acoustic metrics and perceptual judgments were used in correlation analyses to facilitate interpretation of the derived dimensions. Six clusters of similar-sounding speakers and 3 perceptual dimensions underlying similarity were revealed. The clusters of similar-sounding speakers were not constrained by dysarthria subtype diagnosis. The 3 perceptual dimensions revealed by MDS were correlated with metrics for articulation rate, intelligibility, and vocal quality, respectively. This study shows (a) feasibility of a free-classification approach for studying perceptual similarity in dysarthria, (b) correspondence between acoustic and perceptual metrics to clusters of similar-sounding speakers, and (c) similarity judgments transcended dysarthria subtype diagnosis.


Experimental nephropathy induced by human polyclonal light chains.
We describe the nephrotoxic effects of a preparation of light chains (LC) obtained by reduction and alkylation of human IgG. The acute renal lesions in rats are dependent upon the dose and route of LC administration. The kidney alterations were not observed in animals treated with comparable amounts of human albumin or the F(ab')2 fragment of human IgG. Intravenous bolus injection of 120 mg LC into 80-100 g hydropenic rats induced extensive cast formation in distal and collecting renal tubules, which were similar to the human "myeloma kidney". In contrast, the slow infusion of the same amount of LC over 1 h produced degenerative changes in the proximal tubular cells without extensive cast formation. The morphological alterations of the kidney were investigated by classical histological methods, by immunofluorescence and in thin sections stained with toluidine blue. Urinary excretion and kidney content of the lysosomal enzyme N-acetyl-beta-D-glucosaminidase were increased only in the group of animals infused with LC. These findings may be relevant to the pathogenesis of human nephropathy occurring in multiple myeloma and in some autoimmune diseases.


Malaria treatment in the retail sector: knowledge and practices of drug sellers in rural Tanzania.
Throughout Africa, the private retail sector has been recognised as an important source of antimalarial treatment, complementing formal health services. However, the quality of advice and treatment at private outlets is a widespread concern, especially with the introduction of artemisinin-based combination therapies (ACTs). As a result, ACTs are often deployed exclusively through public health facilities, potentially leading to poorer access among parts of the population. This research aimed at assessing the performance of the retail sector in rural Tanzania. Such information is urgently required to improve and broaden delivery channels for life-saving drugs. During a comprehensive shop census in the districts of Kilombero and Ulanga, Tanzania, we interviewed 489 shopkeepers about their knowledge of malaria and malaria treatment. A complementary mystery shoppers study was conducted in 118 retail outlets in order to assess the vendors' drug selling practices. Both studies included drug stores as well as general shops. Shopkeepers in drug stores were able to name more malaria symptoms and were more knowledgeable about malaria treatment than their peers in general shops. In drug stores, 52% mentioned the correct child-dosage of sulphadoxine-pyrimethamine (SP) compared to only 3% in general shops. In drug stores, mystery shoppers were more likely to receive an appropriate treatment (OR = 9.6), but at an approximately seven times higher price. Overall, adults were more often sold an antimalarial than children (OR = 11.3). On the other hand, general shopkeepers were often ready to refer especially children to a higher level if they felt unable to manage the case. The quality of malaria case-management in the retail sector is not satisfactory. Drug stores should be supported and empowered to provide correct malaria-treatment with drugs they are allowed to dispense. At the same time, the role of general shops as first contact points for malaria patients needs to be re-considered. Interventions to improve availability of ACTs in the retail sector are urgently required within the given legal framework.


Knockdown of PKM2 induces apoptosis and autophagy in human A549 alveolar adenocarcinoma cells.
The M2 isoform of pyruvate kinase (PKM2), which has been identified as the predominant cause of the Warburg effect in cancer cells, is essential in tumor metabolism and growth. However, the role of PKM2 in autophagy remains to be elucidated. The present study investigated the effect of PKM2 knockdown on autophagy and apoptotic cell death in human A549 alveolar adenocarcinoma cells. Two short hairpin (sh)RNAs targeting human PKM2 mRNA were designed and lentiviral vectors were constructed. The A549 cells were infected with lentiviruses, containing shRNAs against PKM2, and the expression of PKM2 was examined by reverse transcription-quantitative polymerase chain reaction (RT‑qPCR) and immonoblotting analysis. A lactose dehydrogenase (LDH)‑coupled enzyme assay was used to detect the pyruvate kinase activity. RT‑qPCR was used to detect the mRNA expression level of glycolysis‑associated enzymes. The quantification of cells with punctate LC3 and expression of LC3II were examined to demonstrate autophagy. An MTT assay was used to detect cell viability and flow cytometry was used to determine cell apoptosis. The activity of caspase 3/7 and the expression of Bcl‑2 were also detected in A549 cells with PKM2 knockdown. The present study demonstrated that the two shRNAs efficiently downregulated the mRNA and protein expression levels of PKM2 in A549 cells. The knockdown of PKM2 decreased pyruvate kinase activity and glycolysis. Autophagy was induced in A549 cells with PKM2 knockdown. Inhibition of autophagy accelerated apoptotic death in PKM2‑knockdown cells and this was dependent on increased caspase 3/7 activity and decreased expression of Bcl‑2. In conclusion, the downregulation of PKM2 induced apoptosis and autophagy in A549 cells and this autophagy protected the cells from apoptotic cell death.


Determination of volatile components in ginger using gas chromatography-mass spectrometry with resolution improved by data processing techniques.
Ginger is widely used as either a food product or an herbal medicine in the world. In this paper, a method was developed for determining volatile components in essential oils from both dried and fresh ginger by use of gas chromatography-mass spectrometry (GC-MS) and chemometric approaches. With the resolution improvement by chemometric methods upon two-dimensional data from GC-MS, the drifting baseline can be corrected. In addition, the peak purity can be assessed and the number of chemical components and their stepwise elution in the peak clusters can be identified. The peak clusters investigated are then resolved into pure chromatograms and related mass spectra for each of the components involved. Finally, with the pure chromatograms and related mass spectra obtained, the chemical components can be qualitatively identified based on the similarity searches in the MS databases and the chromatographic retention times. Quantitative determination can be conducted using the overall volume integration approach. The results showed that 140 and 136 components were separated and that 74 and 75 of them were tentatively identified, which accounted for about 62.82 and 47.11% of the total relative content for dried and fresh ginger, respectively. In comparison with the chromatographic fingerprints of essential oils from dried and fresh ginger, 60 of the volatile components determined match with each other. The study demonstrated that the use of chemometric resolution based on two-dimensional data can mathematically enhance the separation ability of GC-MS and assist qualitative and quantitative determination of chemical components separated from complicated practical systems such as foods, herbal medicines, and environmental samples.


Biocompatibility and resorption of a brushite calcium phosphate cement.
A hydraulic calcium phosphate cement with beta-tricalcium phosphate (TCP) granules embedded in a matrix of dicalcium phosphate dihydrate (DCPD) was implanted in experimentally created defects in sheep. One type of defect consisted of a drill hole in the medial femoral condyle. The other, partial metaphyseal defect was located in the proximal aspect of the tibia plateau and was stabilized using a 3.5 mm T-plate. The bone samples of 2 animals each per group were harvested after 2, 4, 6 and 8 weeks. Samples were evaluated for cement resorption and signs of immediate reaction, such as inflammation, caused by the cement setting in situ. Differences regarding these aspects were assessed for both types of defects using macroscopical, radiological, histological and histomorphometrical evaluations. In both defects the brushite matrix was resorbed faster than the beta-TCP granules. The resorption front was followed directly by a front of new bone formation, in which residual beta-TCP granules were embedded. Cement resorption occurred through (i) extracellular liquid dissolution with cement disintegration and particle formation, and (ii) phagocytosis of the cement particles through macrophages. Signs of inflammation or immunologic response leading to delayed new bone formation were not noticed at any time. Cement degradation and new bone formation occurred slightly faster in the femur defects.


[Maternal mortality at the Gynecologic-Obstetric Hospital of the Western National Medical Center, Mexico. A 12-year review].
The aim of this study is to present the analysis of the leading causes of mortality found in all the cases of maternal death at the Hospital de Ginecobstetricia (HGO) and Hospital de Especialidades (HE) del Centro Médico Nacional de Occidente (CMNO), during a period of 12 years, to achieve a winder view and to find a solution to reduce maternal mortality (MM). We reviewed files and reports of the maternal mortality committee of 151 women who were hospitalized and died at the HGO and HE del CMNO in Gudalajara, Jalismo, México, between January first, 1985 and December 31, 1996. During this period the maternal death ratio was 64 per 100,000 live births. The main cause of maternal death was toxemia 25%, follow by hemorrhage 23%, pulmonary thromboembolism 16%, infection 13%, and anesthetic complications 6%. 73% were foreseen deaths and 23% were unforeseen deaths. At the patients arrival to the hospital 37% were avoidable deaths. 66% were direct obstetric deaths and 34% were avoidable deaths. 66% were direct obstetric deaths and 34% were indirects. In 50% of the cases the end of pregnancy was done by cesarean section. The professional responsibility existed in the 72% and the hospitalary in the 23%. We made an analysis related factors and we propose strategies to diminish the maternal mortality.


Hydrocortisone/cyclodextrin complex electrospun nanofibers for a fast-dissolving oral drug delivery system.
The electrospinning of hydrocortisone/cyclodextrin complex nanofibers was performed in order to develop a fast-dissolving oral drug delivery system. Hydrocortisone is a water-insoluble hydrophobic drug, yet, the water solubility of hydrocortisone was significantly enhanced by inclusion complexation with hydroxypropyl-beta-cyclodextrin (HP-β-CyD). In this study, hydrocortisone/HP-β-CyD complexes were prepared in aqueous solutions having molar ratios of 1/1, 1/1.5 and 1/2 (hydrocortisone/HP-β-CyD). Highly concentrated aqueous solutions of HP-β-CyD (180%, w/v) were used for hydrocortisone/HP-β-CyD systems (1/1, 1/1.5 and 1/2) in order to perform electrospinning without the use of an additional polymer matrix. The turbidity of hydrocortisone/HP-β-CyD (1/1 and 1/1.5) aqueous solutions indicated the presence of some uncomplexed crystals of hydrocortisone whereas the aqueous solution of hydrocortisone/HP-β-CyD (1/2) was homogeneous indicating that hydrocortisone becomes totally water-soluble by inclusion complexation with HP-β-CyD. Nonetheless, the electrospinning of hydrocortisone/HP-β-CyD systems (1/1, 1/1.5 and 1/2) successfully yielded defect-free uniform nanofibrous structures. Moreover, the electrospinning process was quite efficient that hydrocortisone was completely preserved without any loss yielding hydrocortisone/HP-β-CyD nanofibers having the initial molar ratios (1/1, 1/1.5 and 1/2). The structural and thermal characterization of the hydrocortisone/HP-β-CyD nanofibers revealed that hydrocortisone was totally inclusion complexed with HP-β-CyD and was in the amorphous state in hydrocortisone/HP-β-CyD (1/2) nanofibers whereas some uncomplexed crystalline hydrocortisone was present in hydrocortisone/HP-β-CyD (1/1 and 1/1.5) nanofibers. Nevertheless, hydrocortisone/HP-β-CyD (1/1, 1/1.5 and 1/2) complex aqueous systems were electrospun in the form of nanofibrous webs having a free-standing and flexible nature. The hydrocortisone/HP-β-CyD (1/1, 1/1.5 and 1/2) nanofibrous webs have shown fast-dissolving behavior in water or when they were in contact with artificial saliva. Yet, the hydrocortisone/HP-β-CyD (1/2) nanofibrous web dissolved more quickly than the hydrocortisone/HP-β-CyD (1/1 and 1/1.5) nanofibrous webs due to the full inclusion complexation and the amorphous state of hydrocortisone in this sample. In short, the results suggest that polymer-free electrospun nanofibrous webs produced from hydrocortisone/HP-β-CyD could be quite applicable for fast-dissolving oral drug delivery systems.


Targeting Fatty-Acid Amide Hydrolase with Prodrugs for CNS-Selective Therapy.
The blood-brain barrier (BBB) can be a substantial impediment to achieving therapeutic levels of drugs in the CNS. Certain chemical functionality such as the carboxylic acid is a general liability for BBB permeability preventing significant CNS distribution of a drug from a systemic dose. Here, we report a strategy for CNS-selective distribution of the carboxylic acid containing thyromimetic sobetirome using prodrugs targeted to fatty-acid amide hydrolase (FAAH), which is expressed in the brain. Two amide prodrugs of sobetirome were shown to be efficient substrates of FAAH with Vmax/KM values comparable to the natural endocannabinoid FAAH substrate anandamide. In mice, a systemic dose of sobetirome prodrug leads to a substantial ∼60-fold increase in brain distribution (Kp) of sobetirome compared to an equimolar systemic dose of the parent drug. The increased delivery of sobetirome to the brain from the prodrug was diminished by both pharmacological inhibition and genetic deletion of FAAH in vivo. The increased brain exposure of sobetirome arising from the prodrug corresponds to ∼30-fold increased potency in brain target engagement compared to the parent drug. These results suggest that FAAH-targeted prodrugs can considerably increase drug exposure to the CNS with a concomitant decrease in systemic drug levels generating a desirable distribution profile for CNS acting drugs.


Imaging of digital neuromas.
In this case presentation, an incision was made on the nonweightbearing surface of the hallux directly over the mass in question, just long enough to allow for the isolation of the entire mass. This permitted easier identification of the mass and enabled dissection of the abnormal tissue and excision of only the tumor with a minimum of tissue trauma. Healing was uneventful and expedient largely because of the reduced tissue handling. Prior to the advent of magnetic resonance imaging, this type of preoperative detailed surgical mapping would not have been possible. Continuing improvements in magnetic resonance imaging hold great and increasing promise.


Prenatal glucocorticoid exposure in the sheep alters renal development in utero: implications for adult renal function and blood pressure control.
Treatment of the pregnant ewe with glucocorticoids early in pregnancy results in offspring with hypertension. This study examined whether glucocorticoids can reduce nephron formation or alter gene expression for sodium channels in the late gestation fetus. Sodium channel expression was also examined in 2-mo-old lambs, while arterial pressure and renal function was examined in adult female offspring before and during 6 wk of increased dietary salt intake. Pregnant ewes were treated with saline (SAL), dexamethasone (DEX; 0.48 mg/h) or cortisol (CORT; 5 mg/h) over days 26-28 of gestation (term = 150 days). At 140 days of gestation, glomerular number in CORT and DEX animals was 40 and 25% less, respectively, compared with SAL controls. Real-time PCR showed greater gene expression for the epithelial sodium channel (α-, β-, γ-subunits) and Na(+)-K(+)-ATPase (α-, β-, γ-subunits) in both the DEX and CORT group fetal kidneys compared with the SAL group with some of these changes persisting in 2-mo-old female offspring. In adulthood, sheep treated with dexamethasone or cortisol in utero had elevated arterial pressure and an apparent increase in single nephron glomerular filtration rate, but global renal hemodynamics and excretory function were normal and arterial pressure was not salt sensitive. Our findings show that the nephron-deficit in sheep exposed to glucocorticoids in utero is acquired before birth, so it is a potential cause, rather than a consequence, of their elevated arterial pressure in adulthood. Upregulation of sodium channels in these animals could provide a mechanistic link to sustained increases in arterial pressure in cortisol- and dexamethasone-exposed sheep, since it would be expected to promote salt and water retention during the postnatal period.


Successful Pupation of Small Hive Beetle, Aethina tumida (Coleoptera: Nitidulidae), in Greenhouse Substrates.
The small hive beetle, Aethina tumida Murray, is an invasive pest that has spread globally. Western honey bees, Apis mellifera Linnaeus (Hymenoptera: Apidae), are considered the most important host and infestations can lead to collapse of colonies. Larvae feed on honey, pollen, and brood inside the hive and leave the hive as postfeeding wandering larvae to pupate in the surrounding soil. Other host species include bumble bees, stingless bees, and solitary bees, all of which can facilitate small hive beetle reproduction and are used for greenhouse crop pollination worldwide. Here, we investigated if small hive beetles can complete their life cycle when soil is absent by pupating in plant root-supporting substrates commonly used in greenhouses. Wandering small hive beetle larvae were introduced into containers with coconut fiber, perlite, a mixture of both and stone wool substrates to investigate pupation success and development time. Sand was used as control substrate. In all but one substrate (perlite), small hive beetles developed into adults equally well as they did in the sand. Development time ranged between 23 and 37 d and was not different from that of the control. We showed that small hive beetles can pupate in greenhouse substrates. This could constitute a problem for greenhouse pollination as well as it could facilitate small hive beetle survival in areas which otherwise would be deemed unsuitable or marginal environments for small hive beetles to become established. Our study highlights the opportunistic nature of the small hive beetle as an invasive species.


[Relative role of physical inactivity and snacking between meals in weight gain].
Diet and life-styles are considered as the main factors that determine the high prevalence of obesity in Western societies. Although some countries have registered a decrease in fat intake, the percentage of overweight and obesity has increased. Therefore, it is thought that fat intake may not be the main factor that determines the current epidemic of obesity. The objective of this study was to determine the role of a sedentary life-style and eating between meals (snacking) as major determinants of a recent weight change (over last 5 years). By using cross-sectionally baseline data of the SUN cohort, we adjusted non-conditional logistic regression models to estimate the odds ratio (OR) of gaining weight according to age, physical activity in leisure time, watching television, taking a nap, smoking, snacking and the intake of macronutrients. A statistically significant inverse association between leisure-time physical activity and the probability of gaining weight was found for men (OR = 0.93; CI 95%, 0.88-0.98) and a trend was also present among women. Snacking was positively associated with a higher probability of gaining weight among men (OR = 1.88; CI 95%, 1.40-2.53) and among women (OR = 1.38; CI 95%, 1.10-1.73). Our data suggest a direct association between snacking and weight gain in middle-aged people.


[Blood serum cytokines as predictors of hemorrhagic transformation of ischemic stroke].
Objectives. Authors studied the levels of pro- (IL-1Β, TNF-α, IL-6, IL-8, IFN-γ) and anti-inflammatory cytokines (IL-1Ra, IL-10, IL-4) in the blood serum at 1st and 2nd day of acute period of ischemic stroke with hemorrhagic transformation (IS HT). Material and methods. We compared cytokines levels in the blood serum of patients with IS HT and in patients with ischemic (IS) and hemorrhagic stroke (HT). Results. In patients with IS HT, significantly lower levels of IL-1Β, TNO-α and higher levels of IL-6 were noted at 1st and 2nd day compared to patients with IS and HS. Significantly lower levels of IFN-γ and IL-10 with increased levels of IL-4 were noted at 1st and 2nd day of IS HT compared to IS. There were no differences in concentrations of IFN-γ, IL-10 and IL-4 between IS HT and HS groups. Conclusion. Cytokines are not only involved in the development of HT in IS but can be predictors of HT.