Reducing ethnic disparity in access to high-quality HLA-matched cord blood units for transplantation: analysis of the Canadian Blood Services' Cord Blood Bank inventory.
Launched in 2013, Canadian Blood Services' Cord Blood Bank (CBS' CBB) has built a high-quality, ethnically diverse cord blood repository that aims to reduce ethnic disparity in accessing suitable units for transplantation. As of December 2016, 2000 units have been banked. The self-reported maternal ethnicity was 58% non-Caucasian. Overall, 26% of units were classified as multi-ethnicity with Caucasian (84%) most frequently observed in combination with Asian, First Nations (predominant indigenous peoples in Canada south of the Arctic Circle), or African ethnicity. Utilization scores that incorporate total nucleated and CD34+ cell counts in the CBS' CBB were associated with greater likelihood of utilization compared with the international inventory of units (p < 0.05). The distribution of utilization scores was similar for Caucasians compared with non-Caucasians (p < 0.05). Using HLA genotypes of cord blood units and their mothers, we determined probable ethnic assignments for each haplotype using HaploStats (National Marrow Donor Program). Significant increases in HLA-match likelihoods are predicted for all ethnicities as the inventory grows to its target of 10,000 units and the gap in HLA-match likelihoods for Caucasian and non-Caucasian patients progressively declines. The CBS' CBB inventory is predicted to have high HLA-matching likelihoods across a broad spectrum of ethnic groups, improving access to high-quality stem cell products for all patients.


Involvement of the nucleotide excision repair protein UvrA in instability of CAG*CTG repeat sequences in Escherichia coli.
Several human genetic diseases have been associated with the genetic instability, specifically expansion, of trinucleotide repeat sequences such as (CTG)(n).(CAG)(n). Molecular models of repeat instability imply replication slippage and the formation of loops and imperfect hairpins in single strands. Subsequently, these loops or hairpins may be recognized and processed by DNA repair systems. To evaluate the potential role of nucleotide excision repair in repeat instability, we measured the rates of repeat deletion in wild type and excision repair-deficient Escherichia coli strains (using a genetic assay for deletions). The rate of triplet repeat deletion decreased in an E. coli strain deficient in the damage recognition protein UvrA. Moreover, loops containing 23 CTG repeats were less efficiently excised from heteroduplex plasmids after their transformation into the uvrA(-) strain. As a result, an increased proportion of plasmids containing the full-length repeat were recovered after the replication of heteroduplex plasmids containing unrepaired loops. In biochemical experiments, UvrA bound to heteroduplex substrates containing repeat loops of 1, 2, or 17 CAG repeats with a K(d) of about 10-20 nm, which is an affinity about 2 orders of magnitude higher than that of UvrA bound to the control substrates containing (CTG)(n).(CAG)(n) in the linear form. These results suggest that UvrA is involved in triplet repeat instability in cells. Specifically, UvrA may bind to loops formed during replication slippage or in slipped strand DNA and initiate DNA repair events that result in repeat deletion. These results imply a more comprehensive role for UvrA, in addition to the recognition of DNA damage, in maintaining the integrity of the genome.


Flash vacuum pyrolysis over solid catalysts. 2. pyrazoles over hydrotalcites.
Flash vacuum pyrolysis (fvp) reactions of NH-pyrazole (1) and 3,5-diphenylpyrazole (2) were investigated in the presence of anionic clays having hydrotalcite structure (HT). Solid catalysts with Mg:Al ratio equal to 2:1 containing carbonate (HT-1), nitrate (HT-2), and silicate (HT-3) as interlayer anions were employed. Between 400 and 600 degrees C, compound 1 remained almost unchanged and only unidentified volatile products were detected in small amounts. In contrast, 2 afforded benzonitrile (3) and phenylacetonitrile (4) by a ring fragmentation reaction at 450 degrees C. At a higher temperature (660 degrees C), the same products obtained in homogeneous fvp reactions, i.e., 2-phenylindene (5) and 3-phenylindene (6), were obtained showing no catalysis by HT under these conditions. Results showed that the yield is strongly dependent on the nature of the interlayer anion in the hydrotalcite structure. In comparison with reactions of 2 over zeolites, HTs exhibit selectivity for ring fragmentation reaction.


Chinese hamster ovary (CHO) cell clustering does not correlate with in vivo histamine-sensitization when measuring residual activity of aldehyde-treated pertussis toxin (PT).
CHO cell clustering test failed to detect a reversion to PT toxicity of a commercial DTaP vaccine batch that failed to pass the test for reversion to histamine sensitizing (HS) activity. Efficacy of CHO cell clustering assay was, accordingly, evaluated using purified PT treated with graded concentrations of formaldehyde at 37 degrees C for 24h. The formaldehyde-treated PT was shown to lose CHO cell clustering activity to the level of 0.01% by the mild treatment while retaining 3.7-20.3% of HS activity which were far over the levels of commercial vaccines. When a reversion to toxicity of the aldehyde-detoxified PT was examined by incubating at 37 degrees C for three weeks, CHO cell clustering test again failed to detect the reversion to toxicity of the PT which showed a remarkable reversion to HS activity. These findings suggested that CHO cell clustering test might have an efficacy limitation in predicting in vivo activity of aldehyde-treated PT.


The biosynthesis of leukotriene B4, the predominant lipoxygenase product in rabbit alveolar macrophages, is enhanced during immune activation.
Rabbit alveolar macrophages synthesize prostaglandins in response to various stimuli. We have previously shown that prostaglandin production was decreased in immunologically activated (live Bacillus Calmette-Guérin (BCG)-injected) animals. (Hsueh, W., Lamb, R. and Gonzalez-Crussi, F. (1982) Biochim. Biophys. Acta 710, 406-414). In the present study, we examined the lipoxygenase products of alveolar macrophages from normal and immunologically activated rabbits injected intravenously with live BCG or complete Freund's adjuvant. We found: unstimulated lung macrophages produced no detectable leukotrienes; the predominant lipoxygenase product upon stimulation was leukotriene B4; alveolar macrophages did not significantly degrade leukotriene B4 into its 20-hydroxy derivative, and the total degradation of leukotriene B4 during 90 min of incubation was minimal; the production of leukotriene B4 reached the peak at 30 min after A23187 stimulation, while zymosan caused a much slower and smaller release; following stimulation, immunologically activated lung macrophages produced more leukotriene B4 than resident macrophages. It is possible that the increased leukotriene B4 production in immunologically activated lung macrophages was related to the immunoregulatory function of this substance, such as enhancing cytotoxicity, interferon production and proliferation of suppressor-cytotoxic T cells.


[Renal hypothermia in situ. Comparison between surface and perfusion cooling concerning renal function in pigs (author's transl)].
There are 2 competing methods for cooling the kidney in situ during surgical ischemia: from without by applying ice to the renal surface and from within by perfusing the renal artery. The latter procedure is said to be superior in protecting renal function. Herein the protective effect on renal function of both methods are compared. Pigs of 15--25 kg weight underwent nephrectomy on one side. The remaining kidney was subjected to cold ischemia during 90 minutes while perfusion- or surface cooling was performed. For perfusion cooling the aorta was punctured and the catheter introduced into the renal artery. The perfusing liquid consisted of a physiologic electrolyt solution (Ringer-Lactate) with heparin kept at a temperature of 3--5 degrees C. The initial perfusion lasted 10 minutes and resulted in a median renal core temperature of 23 degrees C. Then the kidney was put on a cooling pad and every 15 minutes again perfused for one minute. For surface cooling sterile melting ice made of glucose solution 5% was applied directly to the kidney. The renal core temperature could be kept at 15--20 degrees C. The two methods of hypothermia were judged by comparing the serum creatinine levels and the I131-hippuran clearances one month after surgery. There was no difference whatever as analysed by the t-test. Hypothermia by applying ice to the renal surface therefore proved to be equivalent to hypothermia by perfusion. Moreover it is much simpler.


Molecular mechanisms of Polyphyllin I-induced apoptosis and reversal of the epithelial-mesenchymal transition in human osteosarcoma cells.
Osteosarcoma is a most common highly malignant bone tumor in children and adolescents. Polyphyllin I (PPI) is an ethanol extraction from Paris polyphylla Smith var.yunnanensis (Franch.) Hand.-Mazz, which belongs to antipyretic-detoxicate family and has been used as a natural medicine in the treatment of infectious disease and cancer in China for centuries. The proteasome activity inhibitory and anti-osteosarcoma effects of PPI have not been known. Here we found PPI exhibited a selective inhibitory effect on proteasomal chymotrypsin (CT)-like activity, both in purified human proteasome and in cultured osteosarcoma cellular proteasome, and caused an accumulation of ubiquitinated proteins. PPI also inhibited viability, proliferation, migration, and invasion of MG-63, Saos-2, and U-2 OS osteosarcoma cells and resulted in S phase arrest and apoptosis. Furthermore, we explored the molecular targets involved. Exposure of osteosarcoma cells to PPI caused an inactivation of the intrinsic nuclear factor κB (NF-κB) and activation of unfolded protein response (UPR)/endoplasmic reticulum (ER) stress signaling cascade in osteosarcoma cells, followed by down-regulation of anti-apoptotic proteins, with up-regulation of pro-apoptotic proteins. We also demonstrated down-regulation of c-Myc, Cyclin B1, Cyclin D1, and CDK1, which are involved in the cell cycle and growth. Finally, we identified down-regulation of Vimentin, Snail, Slug, and up-regulation of E-cadherin, which are integral proteins involved in epithelial-mesenchymal transition (EMT). Taken together, our data provide insights into the mechanism underlying the anticancer activity of PPI in human osteosarcoma cells.


The active site of the thioredoxin-like domain of chloroplast protein disulfide isomerase, RB60, catalyzes the redox-regulated binding of chloroplast poly(A)-binding protein, RB47, to the 5' untranslated region of psbA mRNA.
RB60, a chloroplast protein disulfide isomerase, modulates the binding of RB47, chloroplast poly(A)-binding protein, to the 5'-UTR of the psbA mRNA using redox potential, allowing for a reversible switch capable of regulating psbA mRNA translation in a light/dark dependent manner. RB60 contains two thioredoxin-like domains with putative catalytic sites of -Cys-Gly-His-Cys- that are presumed to function as active sites for the redox-regulated changes in RNA-binding activity of RB47. To investigate whether these motifs are required for redox-regulated RNA binding, RNA-gel-mobility shift assays were performed with RB47 and mutant RB60 proteins with single cysteines changed to serines in the -Cys-Gly-His-Cys- motif. The results showed that each thioredoxin-like domain has independent catalytic function in the reactivation of RB47 binding and that a double active site mutant completely lacks the ability to activate RB47 RNA binding activity.


Vernal keratoconjunctivitis presenting unilaterally.
A 7-year-old boy had itching, foreign body sensation, and redness in his right eye. Unilateral cobblestone papillae and a shield ulcer were found. Topical antihistamines, mast cell stabilizers, and steroids led to marked improvement. Unilateral vernal keratoconjunctivitis should be included in the differential diagnosis of unilateral giant papillary conjunctivitis.


Aprotinin induced lipohypertrophy and glomerulonephritis in an insulin dependent diabetic.
In an insulin dependent diabetic who was hyperglycaemic and ketotic despite 3,000 u of insulin injected subcutaneously in 2 divided doses daily, 50 u of intravenous insulin infused over 24 hr restored normal glucose homeostasis. A combination of insulin (800 u) and aprotinin (10,000 u) given twice daily also produced adequate glucose homeostasis for a period of 12 months. The patient then developed local hypertrophy of subcutaneous tissue at the injection site and her diabetic control deteriorated. Non-selective proteinuria followed and she developed nephrotic syndrome. Renal biopsy revealed a membraneous glomerulonephritis with subepithelial immune complexes, appearances consistent with a drug-induced glomerulonephritis. Withdrawal of aprotinin led to a gradual remission of nephrotic syndrome and proteinuria over several months. During this period, her diabetes was well controlled with continuous subcutaneous infusion of insulin at a dose of 500 u/24 hr. This case report demonstrates: the effective use of aprotinin for prolonged periods in insulin dependent diabetics with abnormal absorption of subcutaneously injected insulin; aprotinin induced lipohypertrophy which was not observed when insulin was injected alone; aprotinin-associated glomerulonephritis and nephrotic syndrome; the effective use of CSII--at higher insulin doses--in such patients with subcutaneous malabsorption of insulin.


[An expedient synthesis of fluorescent labeled ceramide-1-phosphate analogues].
A synthesis for fluorescent analogs of ceramide-1-phosphate bearing 9-anthrylvinyl or 4,4-difluoro-3a,4a- diaza-s-indacene-8-yl (Me4-BODIPY) fluorophore at o-position of fatty acid residue was carried out. The key stage of the synthesis is hydrolysis of corresponding sphingomyelins catalyzed by phospholipase D from Streptomyces chromofuscus; the enzymatic yield has been raised to 50-70% by appliance of organic solvent in the incubation medium.


Phase shifting of the circadian conidiation rhythm in Neurospora crassa by calmodulin antagonists.
The effects of chemicals capable of antagonizing the functions of calmodulin, such as trifluoperazine, chlorpromazine, imipramine, alprenolol, W7, and W13, on the circadian conidiation rhythm of Neurospora crassa were examined. Trifluoperazine, at a 30-microM concentration, was most effective in shifting the phase of the conidiation rhythm and caused a maximum phase delay at circadian time (CT) 6 and maximum phase advance at CT 9. Chlorpromazine was less effective than trifluoperazine, and a 300-microM concentration of chlorpromazine was required for a similar phase shift. Imipramine, at a 1-mM concentration, caused only a small phase shift, while alprenolol had little effect on biological clock function. W7 and W13 caused phase delays longer than 10 hr at CT 6 and caused a phase advance of about 5 hr at CT 10 when present at a 200-microM concentration. However, W5 and W12, the dechlorinated homologues of W7 and W13, had no effects on clock function at the same concentration. Calmodulin was assayed by measurements of stimulation of cyclic nucleotide diphosphodiesterase activity. Calmodulin content remained constant in trifluoperazine-sensitive and trifluoperazine-insensitive phases for two cycles following the light-dark transition.


Tumor-directed targeting of liposomes.
Tumor-specific targeting is a critical goal in the research area of liposomal drug delivery. Identification of the specific interactions between ligands and target tumor cells is a principle prerequisite in achieving this goal. Generally, tumor cells aberrantly express tumor-associated antigens that can be utilized as appropriate target molecules. Monoclonal antibodies against tumor-associated antigens have been successfully adopted for targeting to various types of cancer cells. The incorporation of humanized monoclonal antibodies or single chain human antibodies, instead of rodent antibodies into immunoliposomes has resulted in better clinical applicability. Tumor-specific ligands other than monoclonal antibodies have also been investigated as in vivo tumor-directing molecules. However, the number of pre-clinical studies of anticancer treatments using tumor-specific liposomal drugs reporting successful targeting and enhanced therapeutic efficacy has been limited. Further refinement of tumor-specific interactions and liposomal formulations will be necessary for the application of the tumor-specific liposomal drug strategy for anticancer chemotherapy or gene therapy.


Antigenic properties of the human immunodeficiency virus transmembrane glycoprotein during cell-cell fusion.
Human immunodeficiency virus (HIV) entry is triggered by interactions between a pair of heptad repeats in the gp41 ectodomain, which convert a prehairpin gp41 trimer into a fusogenic three-hairpin bundle. Here we examined the disposition and antigenic nature of these structures during the HIV-mediated fusion of HeLa cells expressing either HIV(HXB2) envelope (Env cells) or CXCR4 and CD4 (target cells). Cell-cell fusion, indicated by cytoplasmic dye transfer, was allowed to progress for various lengths of time and then arrested. Fusion intermediates were then examined for reactivity with various monoclonal antibodies (MAbs) against immunogenic cluster I and cluster II epitopes in the gp41 ectodomain. All of these MAbs produced similar staining patterns indicative of reactivity with prehairpin gp41 intermediates or related structures. MAb staining was seen on Env cells only upon exposure to soluble CD4, CD4-positive, coreceptor-negative cells, or stromal cell-derived factor-treated target cells. In the fusion system, the MAbs reacted with the interfaces of attached Env and target cells within 10 min of coculture. MAb reactivity colocalized with the formation of gp120-CD4-coreceptor tricomplexes after longer periods of coculture, although reactivity was absent on cells exhibiting cytoplasmic dye transfer. Notably, the MAbs were unable to inhibit fusion even when allowed to react with soluble-CD4-triggered or temperature-arrested antigens prior to initiation of the fusion process. In comparison, a broadly neutralizing antibody, 2F5, which recognizes gp41 antigens in the HIV envelope spike, was immunoreactive with free Env cells and Env-target cell clusters but not with fused cells. Notably, exposure of the 2F5 epitope required temperature-dependent elements of the HIV envelope structure, as MAb binding occurred only above 19 degrees C. Overall, these results demonstrate that immunogenic epitopes, both neutralizing and nonneutralizing, are accessible on gp41 antigens prior to membrane fusion. The 2F5 epitope appears to depend on temperature-dependent elements on prefusion antigens, whereas cluster I and cluster II epitopes are displayed by transient gp41 structures. Such findings have important implications for HIV vaccine approaches based on gp41 intermediates.


Lymphocyte calcium signaling from membrane to nucleus.
Ca(2+) signals control a variety of lymphocyte responses, ranging from short-term cytoskeletal modifications to long-term changes in gene expression. The identification of molecules and channels that modulate Ca(2+) entry into T and B lymphocytes has both provided details of the molecular events leading to immune responses and raised controversy. Here we review studies of the pathways that allow Ca(2+) entry, the function of Ca(2+) in the regulation of cell polarity and motility and the principles by which Ca(2+)-dependent transcription regulates lymphocyte function.


Congenital fascial dystrophy--a noninflammatory disease of fascia: the stiff skin syndrome.
Our patient's disease was similar to the persons with stiff skin syndrome described by Esterly and McKusick (1). Stony-hard indurations of the skin and deeper tissue were generalized but most pronounced in the buttocks, thighs, and legs, with limitation of joint mobility and particularly extensive contractures in the lower limbs. The disease was noticed when the patient was 18 months old, and was nonprogressive within a follow-up period of 12 years. There was no visceral involvement except functional impairment of the lungs, probably due to thickened thoracic fascia. Biochemical, histologic, and electron microscopic studies of the skin and muscle were not remarkable. In skin fibroblasts, collagen synthesis was increased and was accompanied by elevated activity of the prolylhydroxylase and lysylhydroxylase, whereas the transferases were not altered. The fascia was considerably thickened, but contained no inflammatory infiltrates. The significant electron microscopic finding was the presence of amianthoid-like collagen fibers in the fascia.


Restrictive and diversifying elements of the anti-myelin/oligodendrocyte glycoprotein antibody response in primate experimental allergic encephalomyelitis.
Autoantibody responses against conformational epitopes of myelin/oligodendrocyte glycoprotein (MOG) possess myelin destructive potential, as demonstrated in the marmoset model of human multiple sclerosis (MS) and in some rodent models of experimental allergic encephalomyelitis. We have previously characterized monoclonal Fab fragments specific for conformational epitopes of MOG that were derived from a combinatorial antibody library generated from a MOG-immune marmoset. In this paper, we address the molecular heterogeneity of humoral responses against MOG in this outbred model of MS by studying additional antibody clones derived from a genetically unrelated animal. We find that all MOG-specific IgGkappa Fab fragments, unrelated to genetic make-up, utilize a restricted set of variable region genes, IGHV1 and IGHV3 for the H chain and IGKV1, IGKV3, and IGKV5 for the L chain. Despite these restricting factors, diversity within these antibody repertoires can be observed, predominantly within the H-chain CDR3 regions. Our findings suggest that only a limited set of Ig genes is necessary to launch a diverse, destructive humoral immune response against a single CNS antigen in primates. These results are the first to contribute to a better understanding of how myelin-directed and potentially destructive autoantibody responses may develop in human MS.


Safety-net versus private hospital setting for brain metastasis patients treated with radiosurgery alone: Disparities in follow-up care and outcomes.
Stereotactic radiosurgery (SRS) alone is an increasingly accepted treatment for brain metastases, but it requires adherence to frequently scheduled follow-up neuroimaging because of the risk of distant brain metastasis. The effect of disparities in access to follow-up care on outcomes after SRS alone is unknown. This retrospective study included 153 brain metastasis patients treated consecutively with SRS alone from 2010 through 2016 at an academic medical center and a safety-net hospital (SNH) located in Los Angeles, California. Outcomes included neurologic symptoms, hospitalization, steroid use and dependency, salvage SRS, salvage whole-brain radiotherapy, salvage neurosurgery, and overall survival. Ninety-three of the 153 patients were private hospital (PH) patients, and 60 were SNH patients. The median follow-up time was 7.7 months. SNH patients received fewer follow-up neuroimaging studies (1.5 vs 3; P = .008). In a multivariate analysis, the SNH setting was a significant risk factor for salvage neurosurgery (hazard ratio [HR], 13.65; P < .001), neurologic symptoms (HR, 3.74; P = .002), and hospitalization due to brain metastases (HR, 6.25; P < .001). More clinical visits were protective against hospitalizations due to brain metastases (HR, 0.75; P = .002), whereas more neuroimaging studies were protective against death (HR, 0.65; P < .001). SNH patients with brain metastases treated with SRS alone had fewer follow-up neuroimaging studies and were at higher risk for neurologic symptoms, hospitalization for brain metastases, and salvage neurosurgery in comparison with PH patients. Clinicians should consider the practice setting and patient access to follow-up care when they are deciding on the optimal strategy for the treatment of brain metastases. Cancer 2018;124:167-75. © 2017 American Cancer Society.


Role of Salmonella enterica serovar Typhimurium two-component system PreA/PreB in modulating PmrA-regulated gene transcription.
The PmrA/PmrB two-component system encoded by the pmrCAB operon regulates the modification of Salmonella enterica serovar Typhimurium lipopolysaccharide leading to polymyxin B resistance. PmrA and PhoP are the only known activators of pmrCAB. A transposon mutagenesis screen for additional regulators of a pmrC::MudJ fusion led to the identification of a two-component system, termed PreA/PreB (pmrCAB regulators A and B), that controls the transcription of the pmrCAB operon in response to unknown signals. The initial observations indicated that insertions in, or a deletion of, the preB sensor, but not the preA response regulator, caused upregulation of pmrCAB. Interestingly, the expression of pmrCAB was not upregulated in a preAB mutant grown in LB broth, implicating PreA in the increased expression of pmrCAB in the preB strain. This was confirmed by overexpression of preA(+) in preAB or preB backgrounds, which resulted in significant upregulation or further upregulation of pmrCAB. No such effect was observed in any tested preB(+) backgrounds. Additionally, an ectopic construct expressing a preA[D51A] allele also failed to upregulate pmrC in any of the pre backgrounds tested, which implies that there is a need for phosphorylation in the activation of the target genes. The observed upregulation of pmrCAB occurred independently of the response regulators PmrA and PhoP. Although a preB mutation led to increased transcription of pmrCAB, this did not result in a measurable effect on polymyxin B resistance. Our genetic data support a model of regulation whereby, in response to unknown signals, the PreB sensor activates PreA, which in turn indirectly upregulates pmrCAB transcription.


Analgesic effect of long-acting somatostatin receptor agonist pasireotide in a patient with acromegaly and intractable headaches.
A 22-year-old woman presented with worsening vision loss and headaches. A diagnosis of acromegaly was confirmed after detection of an invasive pituitary macroadenoma and biochemical testing. Despite two attempts of surgical debulking of the tumour and administration of long-acting octreotide and cabergoline, growth hormone (GH) and insulin-like growth factor 1 (IGF-1) levels were uncontrolled. The patient experienced persistent headaches despite surgery, gamma knife radiation and ventriculoperitoneal shunt placement; she was then enrolled in the ACCESS trial (ClinicalTrials.gov identifier, NCT01995734). Pasireotide (Signifor; Signifor LAR) was initiated, which led to reduced GH and IGF-1 levels and resolution of her intractable headaches. This highlights the use of monthly pasireotide in resolving headaches and improved biochemical control in a patient with acromegaly. We postulate that the headaches improved due to an analgesic and/or anti-inflammatory effect mediated by somatostatin receptors targeted by pasireotide. This may represent an additional benefit of pasireotide and requires further investigation.


Uveitis after COVID-19 Vaccination.
This report describes a case of acute uveitis following COVID-19 vaccination. A 21-year-old patient was referred to our department for decreased visual acuity and injection of the left eye. Two days prior, the patient received her second dose of the severe acute respiratory syndrome coronavirus 2 vaccine (BNT162b2, Pfizer-BioNTech). On ocular examination, her left eye's best-corrected visual acuity was 20/250, and its intraocular pressure was 16 mm Hg on a noncontact tonometer. Hypopyon, 4+ cells, and flares were observed in the anterior chamber. The patient was diagnosed with acute uveitis. She received topical dexamethasone (0.1%) hourly and systemic prednisone (50 mg/day). After 7 days, her inflammatory symptoms were mostly resolved. COVID-19 vaccination can cause acute anterior uveitis, which is responsive to steroids.


Comparison of approaches for efficient gene silencing induced by microRNA-based short hairpin RNA and indicator gene expression.
MicroRNA-based short hairpin RNAs (shRNAs) are natural inducers of RNA interference and have been increasingly used in shRNA expression strategies. In the present study, we compared the efficiencies of exogenous green fluorescence protein (GFP) and endogenous glyceraldehyde-3-phosphate dehydrogenase (GAPDH) knockdown and red fluorescent protein (RFP) indicator expression mediated by three differently designed plasmids. RFP was introduced either at the 5' end, at the 3' end of the human mir155-based target gene (TG) (e.g., GFP or GAPDH) shRNA expression cassette (EC), or at the 3' end of the chimeric intron-containing TG shRNA EC. Comparisons with the control vector showed an obvious reduction of GFP or GAPDH expression with the various shRNA expression plasmids (P < 0.05). When RFP was located at the 5' end or at the 3' end of the TG shRNA EC, RFP expression was low; whereas when RFP was connected with the chimeric intron-containing TG shRNA EC, RFP expression was high. Taken together, this study demonstrated an efficient plasmid design for both TG silencing induced by microRNA-based shRNA and indicator gene expression in vitro.


Fight against background noise in stimulated emission depletion nanoscopy.
STimulated emission depletion (STED) nanoscopy has been proposed to extend greatly our capability of using light to study a variety of biological problems with nanometer-scale resolution. However, in practice the unwanted background noise degrades the STED image quality and precludes quantitative analysis. Here, we discuss the underlying sources of the background noise in STED images, and review current approaches to alleviate this problem, such as time-gating, anti-Stokes excitation removal, and off-focus incomplete depletion suppression. Progress in correcting uncorrelated background photons in fluorescence correlation spectroscopy combined with STED (STED-FCS) will also be discussed.


Benefit of intensified treatment for all children with acute lymphoblastic leukaemia: results from MRC UKALL XI and MRC ALL97 randomised trials. UK Medical Research Council's Working Party on Childhood Leukaemia.
Treatment of children with acute lymphoblastic leukaemia (ALL) aims to cure all patients with as little toxicity as possible and, if possible, to restrict further intensification of chemotherapy to patients with an increased risk of relapse. However in Medical Research Council (MRC) trial UKALL X two short myeloablative blocks of intensification therapy given at weeks 5 and 20 were of benefit to children in all risk groups. The successor trials, MRC UKALL XI and MRC ALL97, tested whether further intensification would continue to benefit all patients by randomising them to receive, or not, an extended third intensification block at week 35. After a median follow-up of 4 years (range 5 months to 8 years), 5 year projected event-free survival was superior at 68% for the 894 patients allocated a third intensification compared with 60% for the 887 patients who did not receive one (odds ratio 0.75, 95% CI 0.63-0.90, 2P = 0.002). This difference was almost entirely due to a reduced incidence of bone marrow relapses in the third intensification arm (140 of 891 in the third intensification arm vs. 171 of 883 in the no third intensification, 2P = 0.02). Subgroup analysis suggests benefit of the third intensification for all risk categories. Overall survival to date is no different in the two arms, indicating that a greater proportion of those not receiving a third intensification arm and subsequently relapsing can be salvaged. These results indicate that there is benefit of additional intensification for all risk subgroups of childhood ALL.


Detector Response and Perfect-Lens-MTF in Polychromatic Light.
Because light sources and detectors exhibit significant spectral variations, the detector response in an optical system can generally be found only by means of a tedious numerical integration process. The evaluation of the MTF in polychromatic light requires an even more extensive numerical procedure. Here, integration results are presented for all combinations of six commonly used light sources and twelve common detectors. The MTF's for perfect lenses and efficiency factors are presented.


Nitrogen ion induced 2D-GaN layer formation of GaAs (001) surface.
This study demonstrates the formation of two-dimensional GaN on GaAs (001) surface by bombardment of nitrogen ions at room temperature. In this work the ion induced nitridation of GaAs (001) surface using nitrogen ion beam of different energies (range from 250 eV to 5 keV) has been investigated using in-situ X-ray Photoelectron Spectroscopy (XPS). A Ga rich surface produced by Art ion etching, promotes initial nitridation. Using nitrogen ion of different energies of constant fluence performs the nitridation. The nitridation suggests that the degree of nitridation increase as the nitrogen ion energy increases up to 3 keV and then attains saturation. The core level and valance band spectra were monitored to observe the chemical and electronic changes as a function of nitrogen ion beam energy. It is observed that Ga(3d) core level peak shifts during nitridation and N(1s) core level spectra shows that the intensity of the nitrogen peak increases and the Ga (LMM) auger peak shifts towards the higher binding energy, reveal the forming of N bonds with Ga by replacing the Ga-As bonds, forming GaN.


Neurons in the primate superior colliculus coding for arm movements in gaze-related coordinates.
In the intermediate and deep layers of the superior colliculus (SC), a well-established oculomotor structure, a substantial population of cells is involved in the control of arm movements. To examine the reference frame of these neurons, we recorded in two rhesus monkeys (Macaca mulatta) the discharges of 331 neurons in the SC and the underlying mesencephalic reticular formation (MRF) while monkeys reached to the same target location during different gaze orientations. For 65 reach-related cells with sufficient data and for simultaneously recorded electromyograms (EMGs) of 11 arm muscles, we calculated an ANOVA (factors: target position, gaze angle) and a gaze-dependency (GD) index. EMGs and the activity of many (60%) of the reach-related neurons were not influenced by the target representation on the retina or eye position. We refer to these as "gaze-independent" reach neurons. For 40%, however, the GD fell outside the range of the muscle modulation, and the ANOVA showed a significant influence of gaze. These "gaze-related" reach neurons discharge only when the monkey reaches for targets having specific coordinates in relation to the gaze axis, i.e., for targets in a gaze-related "reach movement field" (RMF). Neuronal activity was not modulated by the specific path of the arm movement, the muscle pattern that is necessary for its realization or the arm that was used for the reach. In each SC we found gaze-related neurons with RMFs both in the contralateral and in the ipsilateral hemifield. The topographical organization of the gaze-related reach neurons in the SC could not be matched with the well-known visual and oculomotor maps. Gaze-related neurons were more modulated in their strength of activity with different directions of arm movements than were gaze-independent reach neurons. Gaze-related reach neurons were recorded at a median depth of 2.03 mm below SC surface in the intermediate layers, where they overlap with saccade-related burst neurons (median depth: 1.55 mm). Most of the gaze-independent reach cells were found in a median depth of 4.01 mm below the SC surface in the deep layers and in the underlying MRF. The gaze-related reach neurons operating in a gaze-centered coordinate system could signal either the desired target position with respect to gaze direction or the motor error between gaze axis and reach target. The gaze-independent reach neurons, possibly operating in a shoulder- or arm-centered reference frame, might carry signals closer to motor output. Together these two types of reach neurons add evidence to our hypothesis that the SC is involved in the sensorimotor transformation for eye-hand coordination in primates.


[Evapotranspiration of natural Quercus litaotungensis and Tilia paucicostata secondary stands in Liupan Mountains of Ningxia].
With heat dissipation probe technique and combined with microlysimeter and hydrological methods, this paper studied the evapotranspiration of secondary Quercus liaotungensis and Tilia paucicostata stands, and its relationship to forest structure from August to September 2004. The results indicated that the stem sap flux density (SFD) of Quercus liaotungensis and Tilia paucicostata changed regularly from day to night in later growth season. In relatively still period (nighttime), the SFD kept low values continuously, usually below 0.05 microl x cm(-2). min(-1), while in active period (daytime), it increased quickly, usually below 0.25 microl x cm(-2) x min(-1). The daily transpiration of whole-tree was estimated based on the calculation of daily cumulative SFD, which showed that there existed a significant difference between two tree species. The daily transpiration of Quercus liaotungensis reached 5.31 and 2.48 L x d(-1) in sunny- and cloudy-days, 2.35 and 3.75 folds as that of Tilia paucicostata, respectively. There was no significant difference in daily leaf transpiration rate between these two shrub species. During measurement periods, the average daily stand evapotranspiration was 1.45 mm x d(-1), including transpiration (0.72 mm x d(-1)), soil evaporation (0.19 mm x d(-1)), and canopy interception (0.54 mm x d(-1)), which accounted for 49.6%, 13.3% and 37.1% of the total evapotranspiration, respectively. The results showed that the difference of whole-tree transpiration between tree species contributed significantly to the difference of tree canopy transpiration, while the difference of shrub canopy transpiration was on account of the leaf amount in the canopy. The effects of vertical layers in the stand on total stand evapotranspiration also differed, with a percentage of 65.8%, 20.9% and 13.3% for tree canopy, shrub canopy and floor, respectively, which meant that the transpiration and interception of tree canopy contributed most to the total evpotranspiration, followed by shrub transpiration, soil evaporation, and grass transpiration.


Leukemoid reaction in pancreatic cancer: a case report and review of the literature.
The presentation of pancreatic cancer with a leukemoid reaction is rare with no prior reports in the English language literature. We report a case of advanced pancreatic cancer presenting with leukemoid reaction. Granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor levels were normal while interleukin-6 was elevated. The patient had no evidence of infection. The leukemoid reaction correlated with tumor response. This is the first case report in the English literature of a leukemoid paraneoplastic syndrome in a patient with pancreatic cancer. A clear correlation between tumor response, serum carbohydrate antigen 19-9 levels and leukemoid reaction is documented.


Interleukin-4-Pseudomonas exotoxin chimeric fusion protein for malignant glioma therapy.
Human malignant glioma cell lines, primary cell cultures, and tumor specimens derived from surgical samples have been shown to overexpress high-affinity receptors (R) for interleukin-4 (IL-4) in vitro and in situ. The significance of IL-4R expression on malignant glioma cells is still unclear. However, IL-4 has been reported to mediate functional effects in several solid tumor cell lines. These activities include inhibition of cell proliferation, regulation of adhesion molecules, and induction of signal transduction through the JAK/STAT pathway. To target IL-4Rs on tumor cells, we have produced a chimeric recombinant fusion protein consisting of a binding ligand, circularly permuted IL-4 and a mutated form of Pseudomonas exotoxin. This molecule is termed IL4(38-37)-PE38KDEL, cpIL4-PE, or IL-4 cytotoxin. Recombinant cpIL4-PE is highly and specifically cytotoxic to glioma cell lines in vitro, while it is not cytotoxic or less cytotoxic to hematopoietic and normal brain cells. In a nude mouse model, cpIL4-PE showed significant antitumor activity and partial or complete regression of small or large established human glioblastoma tumors. Encouraging preclinical efficacy, safety, and tolerability studies lead to testing of this agent in patients with recurrent glioblastoma. Based on these pilot studies, an extended Phase I/II clinical trial is currently ongoing to determine safety, tolerability, and efficacy of cpIL4-PE when injected stereotactically directly into the recurrent glioma by convection enhanced delivery. Preliminary clinical results suggest that cpIL4-PE can cause pronounced necrosis of recurrent glioma tumors without systemic toxicity. The central nervous system toxicities observed were attributed to the volume of infusion and/or nonspecific toxicity. Ongoing clinical trials will reveal antitumor activities of IL-4 cytotoxin in recurrent malignant glioma.


State of the Academy presidential address 4th scientific conference - Sedona, AZ.
This state of the Academy address was presented at the opening ceremonies of the fourth annual scientific meeting of the American Academy of Health Behavior by the outgoing president of the Academy, February 29, 2004.


The Chinese version of the modification of diet in renal disease (MDRD) equation is a superior screening tool for chronic kidney disease among middle-aged Taiwanese than the original MDRD and Cockcroft-Gault equations.
Three equations have been often used to estimate glomerular filtration rate (GFR), namely, Modification of Diet in Renal Disease (MDRD), MDRD for Chinese (MDRDc), and Cockcroft-Gault (CG), for the purpose of screening individuals with chronic kidney disease (CKD). However, neither of these equations has been tested in a large Asian population. The aim of this study was to determine which equations were suitable for screening CKD in a large Taiwanese population. The applicability of the three equations was analyzed among 32,542 participants of a health examination at Chang Gung Memorial Hospital (CGMH), Taiwan, between 2005 and 2007. Estimated glomerular filtration rate (eGFR)-MDRDc obtained the highest estimate of GFR (mean 101.5 ± 19.2 ml/min/1.73 m 2 ), followed by eGFR-MDRD (mean 83.8 ± 15.8 ml/min/1.73 m 2 ) and eGFR-CG (mean 79.4 ± 29.1 ml/min/1.73 m 2 ). The prevalence of CKD stage 3-5 was 1.9%, 5.1%, and 25.5% according to MDRDc, MDRD and CG equations, respectively. With respect to CKD staging, the agreement between eGFR-MDRDc and eGFR-CG (weighted kappa, k = 0.22) and that between eGFR-MDRD and eGFR-CG (weighted k = 0.30) was poor. Both the original MDRDc and MDRD indicated that subjects with risk factors for CKD had significantly lower eGFR and higher odds ratios for stage 3-5 disease than those without. Paradoxically, the mean eGFR-CG (or odds ratios) was higher (or lower) in subjects with hyperuricemia, hypertension, obesity, or metabolic syndrome than those without these risk factors. The use of the CG equation in the Taiwanese population is inappropriate for screening individuals with CKD, and the MDRDc equation seems to be better for Taiwanese population.


Challenges and opportunities in providing palliative care services to children with a life-limiting illness: A systematic review.
Background Palliative care for children is an innovative approach that helps improve the quality of life of children suffering from life-limiting illnesses, and their family members. The WHO recognized palliative care as a part of universal health coverage. However, there is inadequate availability and inequitable distribution of palliative care services for children in most parts of the world. We reviewed the existing literature to assess (i) the challenges in providing palliative care services for children suffering from life-limiting illnesses and (ii) the strategies or opportunities to overcome these challenges. Methods We conducted systematic searches in the PubMed and Scopus databases to find articles published in the past 10 years (January 2011 to December 2020). The population, concept and context (PCC) framework was used to devise a search strategy in an electronic database. Results A total of 1562 articles were found by searching the database and other sources. Title and abstracts of articles were screened, and 206 articles were selected for full-text review. After scrutiny 28 articles met the inclusion criteria. Barriers to and opportunities in the provision of palliative care services for children were identified at policy, organizational, healthcare provider, and patient/family levels. Conclusion We found that the majority of barriers to provision of palliative care services for children with life-limiting illnesses can be addressed by adopting research-driven strategies. Adequate and equitable distribution of palliative care services is required for improving children and their family members' quality of life.


EP3 (prostaglandin E2 receptor 3) expression is a prognostic factor for progression-free and overall survival in sporadic breast cancer.
In various cancers, overexpression of cyclooxygenase (COX)-2 and elevated prostaglandin (PG) E2 synthesis have been associated with tumor development and progression. The potential of COX-2 inhibitors in cancer prevention and treatment has been shown repeatedly; however, their clinical use is limited due to toxicity. PGE2 signals via EP receptors 1-4, whose functions are analyzed in current research in search for targeted anti-PG therapies. EP2 and EP4 rather promote tumorigenesis, while the role of EP3, especially in breast cancer, is not yet clear and both pro- and anti-tumorigenic effects have been described. Our study evaluates EP3 receptor expression in sporadic breast cancer and its association with clinicopathological parameters, progression-free and overall survival. Two hundred eighty-nine sporadic breast cancer samples without primary distant metastasis were immunohistochemically analyzed for EP3 receptor expression. Tissue was stained with primary anti-EP3-antibodies. Immunoreactivity was quantified by the immunoreactivity-score (IRS); samples with an IRS ≥ 2 scored as EP3 positive. Chi-squared and Mann-Whitney-U test were used for comparison of data; Kaplan-Meier estimates and Cox-regression were used for survival analyses. EP3 receptor was expressed in 205 of 289 samples analyzed (70.9%). EP3 receptor expression was not associated with clinicopathological parameters (e. g. tumor size, hormone receptors, lymph node status). Kaplan-Meier estimates showed a significant association of EP3 positivity with improved progression-free survival (p = 0.002) and improved overall survival (p = 0.001) after up to 10 years. Cox regression analysis confirmed EP3 positivity as a significant prognostic factor even when other known prognosticators were accounted for. In sporadic breast cancer, EP3 receptor expression is not significantly associated with clinicopathological parameters but is a significant prognostic factor for improved progression-free and overall survival. However, the functional aspects of EP3 receptor in breast cancer and the way how EP3 may oppose the pro-tumorigenic effects of PGE2 elevation and COX-2 overexpression are not fully understood so far. Further studies aiming at identification of the factors regulated by EP3 are necessary to evaluate the possibility of targeting EP3 in future anti-tumor therapy in breast cancer.


Assessment of the readability and quality of online patient education materials for the medical treatment of open-angle glaucoma.
Patient adherence to glaucoma medications is poor, and is linked to low literacy levels. Patients commonly use the internet to access health information, and it is recommended that patient information is written at an 11-year-old reading level. The aim of this study is to assess the readability and quality of online patient education materials for the medical management of open angle glaucoma. The top 10 relevant Google searches for nine glaucoma medications (timolol, brimonidine, apraclonidine, dorzolamide, latanoprost, bimatoprost, travoprost, tafluprost and brinzolamide) and three generic searches were analysed for readability and accountability. Readability was assessed using Flesch Reading Ease Score (FRES), Flesch-Kincaid Grade Level (FKGL), Gunning Fog Index (GFI) and Simple Measure of Gobbledygook Index (SMOG). Webpages were classified by source and assessed using Journal of the American Medical Association (JAMA) benchmarks of accountability. 111 articles were included in the analysis. Mean readability scores were: FRES 55.5 (95% CI 53.4 to 57.5); FKGL 9.7 (95% CI 9.3 to 10.0); GFI 12 (95% CI 11.6 to 12.4) and SMOG 9.3 (95% CI 8.9 to 9.6). One-way analysis of variance demonstrated no significant difference in readability score between source type. 9% of the webpages satisfied all 4 JAMA benchmarks. Pearson's correlation coefficient showed a correlation between the FRES and accountability score (r=0.19, p=0.045). The majority of online patient education materials for the medical treatment of glaucoma are written at a level too difficult for the general population and fail to meet accountability standards.


Belief in a just world and subjective well-being: mothers of normal and Down syndrome children.
A cross-sectional study examined the relationship between three dimensions of the belief in a just world and the subjective well-being of Pakistani mothers of normal and Down syndrome children (n = 100 each). Personal belief in a just world and two dimensions of general belief in a just world-beliefs in immanent and ultimate justice-were assessed along with four dimensions of subjective well-being: life satisfaction, mood, state anxiety, and depression. It was hypothesized that personal belief in a just world and belief in ultimate justice would be positively associated with subjective well-being for both groups of mothers. Results were obtained through moderated regression analyses. In line with our hypothesis, personal belief in a just world positively predicted life satisfaction and mood level and negatively predicted state anxiety and depression in both groups of mothers. In contrast, beliefs in immanent and ultimate justice were not consistently adaptive. In particular, the more the mothers of a Down syndrome child believed in immanent justice, the more anxiety they experienced the previous week. In addition, the more the mothers of normal children believed in ultimate justice, the more they experienced anxiety. The pattern of results persisted when controlled for mothers' education, the total number of their children and marital status. Overall, the results support the role of personal belief in a just world as a personal resource in adverse as well as normal life circumstances.


Chairside sensor for rapid monitoring of Enterococcus faecalis activity.
In this study, optical spectroscopy was used to monitor a chromogenic, enzyme-substrate reaction for the rapid identification of Enterococcus faecalis. The detection system, comprising a miniature spectrophotometer and an accompanying data acquisition system, was placed in an incubator. During testing, a 3-ml test sample was placed in a cuvette within the spectrophotometer. This permitted online, real-time, and remote analysis of spectral signature needed to monitor the bacteria. It was observed that the absorption peak intensity increased conspicuously 3.5 h after inoculation and through the entire period of testing. A linear-regression analysis demonstrated a significant correlation between the increase in absorption peak intensity at 610 nm (r = 0.9389) and 653 nm (r = 0.9387) with the formation of colony-forming units. Optical spectroscopy-based sensing systems can pave the way for rapid, nonlaboratory-based approaches to monitor microbial status quantitatively and qualitatively from clinical samples.


Optimum radiographic examination for consideration of compensation awards: II. Cervical and lumbar spines.
In a large Veterans Administration (VA) hospital, a prospective study was undertaken to determine the optimum number of radiographic projections needed to assess applicants for VA compensation for cervical and lumbar spine disease or injury claimed to be service-related. Results show that the final radiographic assessment could be made in 100% (245) of cervical spine examinations using two views (anteroposterior and lateral) and in 99.3% (699/704) of lumbar spine examinations using two views (anteroposterior and a single well centered lateral). Extrapolating these results to data on compensation examinations performed in the entire VA system (fiscal year 1978), it was concluded that limiting the radiographic examinations of the cervical and lumbar spines to these views would eliminate 193,000 radiographs in the evaluation of 100,000 applicants and yield an annual savings of about $1,000,000.


Genetically engineered V79 Chinese hamster cell expression of purified cytochrome P-450IIB1 monooxygenase activity.
Chinese hamster V79 fibroblasts, frequently used as target cells in short-term tests for mutagenicity, do not possess measurable monooxygenase activity; in particular, enzymatic oxidation of testosterone (T) cannot be demonstrated. If these V79 cells, however, had been transfected with the cDNA-encoding rat liver cytochrome P-450IIB1 under control of the SV40 early promoter, they stably expressed monooxygenase activity. These so-called SD1 cells then oxidatively metabolized T at a rate of 27 pmol/mg protein/min, converting it to 16 alpha- and 16 beta-hydroxy-T as well as 4-androsten-3,17-dione as sole metabolites in a ratio of 1.1:1.0:1.6. The regio- and stereoselective conversion of T by SD1 cells, as well as the quantitative distribution of the metabolites, corresponds well with the results reported for pure cytochrome P-450IIB1 in a reconstituted system.


Lab-on-chip cytometry based on magnetoresistive sensors for bacteria detection in milk.
Flow cytometers have been optimized for use in portable platforms, where cell separation, identification and counting can be achieved in a compact and modular format. This feature can be combined with magnetic detection, where magnetoresistive sensors can be integrated within microfluidic channels to detect magnetically labelled cells. This work describes a platform for in-flow detection of magnetically labelled cells with a magneto-resistive based cell cytometer. In particular, we present an example for the validation of the platform as a magnetic counter that identifies and quantifies Streptococcus agalactiae in milk.


Recent advances in amorphous electrocatalysts for oxygen evolution reaction.
Oxygen evolution reaction (OER) has attracted great attention as an important half-reaction in the electrochemical splitting of water for green hydrogen production. However, the inadequacy of highly efficient and stable electrocatalysts has impeded the development of this technology. Amorphous materials with long-range disordered structures have exhibited superior electrocatalytic performance compared to their crystalline counterparts due to more active sites and higher structural flexibility. This review summarizes the preparation methods of amorphous materials involving oxides, hydroxide, phosphides, sulfides, and their composites, and introduces the recent progress of amorphous OER electrocatalysts in acidic and alkaline media. Finally, the existing challenges and future perspectives for amorphous electrocatalysts for OER are discussed. Therefore, we believe that this review will guide designing amorphous OER electrocatalysts with high performance for future energy applications.


Validation and Development of a Modified Breast Graded Prognostic Assessment As a Tool for Survival in Patients With Breast Cancer and Brain Metastases.
Several indices have been developed to predict overall survival (OS) in patients with breast cancer with brain metastases, including the breast graded prognostic assessment (breast-GPA), comprising age, tumor subtype, and Karnofsky performance score. However, number of brain metastases-a highly relevant clinical variable-is less often incorporated into the final model. We sought to validate the existing breast-GPA in an independent larger cohort and refine it integrating number of brain metastases. Data were retrospectively gathered from a prospectively maintained institutional database. Patients with newly diagnosed brain metastases from 1996 to 2013 were identified. After validating the breast-GPA, multivariable Cox regression and recursive partitioning analysis led to the development of the modified breast-GPA. The performances of the breast-GPA and modified breast-GPA were compared using the concordance index. In our cohort of 1,552 patients, the breast-GPA was validated as a prognostic tool for OS (P < .001). In multivariable analysis of the breast-GPA and number of brain metastases (> three v ≤ three), both were independent predictors of OS. We therefore developed the modified breast-GPA integrating a fourth clinical parameter. Recursive partitioning analysis reinforced the prognostic significance of these four factors. Concordance indices were 0.78 (95% CI, 0.77 to 0.80) and 0.84 (95% CI, 0.83 to 0.85) for the breast-GPA and modified breast-GPA, respectively (P < .001). The modified breast-GPA incorporates four simple clinical parameters of high prognostic significance. This index has an immediate role in the clinic as a formative part of the clinician's discussion of prognosis and direction of care and as a potential patient selection tool for clinical trials.


The effect of amiloride on amphotericin B-induced hypokalaemia.
Amiloride was administered to 19 oncology patients exhibiting marked amphotericin B-induced electrolyte wasting. Mean serum potassium concentrations increased in the 5 days preceding and following administration (3.4 +/- 0.5 versus 3.9 +/- 0.8 mmol/L, P = 0.002). A trend towards decreased potassium supplementation was also observed (48.0 +/- 66.5 versus 29.4 +/- 43.2 mmol/day, P = 0.12). Amiloride is a therapeutic option to decrease potassium wasting in patients being treated with amphotericin B.


Hypothalamic-pituitary-adrenal axis activation and stimulation of systemic vasopressin secretion by recombinant interleukin-6 in humans: potential implications for the syndrome of inappropriate vasopressin secretion.
We recently demonstrated that sc administered interleukin-6 (IL-6) strongly stimulates the human hypothalamic-pituitary-adrenal (HPA) axis, with mild toxicity and no hypotensive effects. In this study, we evaluated the response of the human HPA axis to escalating iv doses of recombinant IL-6 in six patients with cancer and good performance status who received daily, every 8 h, three equal doses of 0.3-30 micrograms/kg IL-6. The plasma levels of IL-6 assayed by a specific enzyme-linked immunosorbent assay during the 4 h following the first IL-6 injection were elevated for 2-4 h, proportionally to the amount of injected IL-6. Administration of the cytokine was followed by marked elevations of plasma ACTH (53.0-98.6 pmol/L) and cortisol (824.9-1729.9 nmol/L) independently of the IL-6 dose administered, suggesting that the doses employed were at the top of the dose-response curve for these hormones. Interestingly, plasma arginine vasopressin (AVP) levels were also elevated during the 2 h after IL-6 injection in all patients who received a dose of 3 micrograms/kg or more, suggesting that IL-6 activated the magnocellular AVP-secreting neurons and that it might be involved in the syndrome of inappropriate AVP secretion. Cortisol elevations with peaks similar to those observed after the first injection of IL-6 were also detected in plasma sampled every 2 h after the second and third injections, suggesting that there was no rapid tachyphylaxis in response to IL-6 administration. Plasma IL-1 beta and tumor necrosis factor-alpha concentrations, assayed by specific enzyme-linked immunosorbent assays during the 4 h after the first IL-6 injection, were either within the normal range or undetectable, confirming in vitro observations that IL-6 does not stimulate IL-1 beta or tumor necrosis factor-alpha secretion and suggesting that it exerts its effect on the HPA axis and AVP secretion independently of them. We conclude that IL-6 is a potent stimulator of the human HPA axis and a secretagogue of magnocellular AVP secretion, which might be employed as a challenge test of the axis and the magnocellular AVP neuron.


The nonribosomal peptide synthetase enzyme DdaD tethers N(β)-fumaramoyl-l-2,3-diaminopropionate for Fe(II)/α-ketoglutarate-dependent epoxidation by DdaC during dapdiamide antibiotic biosynthesis.
The gene cluster from Pantoea agglomerans responsible for biosynthesis of the dapdiamide antibiotics encodes an adenylation-thiolation didomain protein, DdaD, and an Fe(II)/α-ketoglutarate-dependent dioxygenase homologue, DdaC. Here we show that DdaD, a nonribosomal peptide synthetase module, activates and sequesters N(β)-fumaramoyl-l-2,3-diaminopropionate as a covalently tethered thioester for subsequent oxidative modification of the fumaramoyl group. DdaC catalyzes Fe(II)- and α-ketoglutarate-dependent epoxidation of the covalently bound N(β)-fumaramoyl-l-2,3-diaminopropionyl-S-DdaD species to generate N(β)-epoxysuccinamoyl-DAP (DAP = 2,3-diaminopropionate) in thioester linkage to DdaD. After hydrolytic release, N(β)-epoxysuccinamoyl-DAP can be ligated to l-valine by the ATP-dependent ligase DdaF to form the natural antibiotic N(β)-epoxysuccinamoyl-DAP-Val.


Biodegradation of methyl tert-butyl ether by a pure bacterial culture.
Biodegradation of methyl tert-butyl ether (MTBE) by the hydrogen-oxidizing bacterium Hydrogenophaga flava ENV735 was evaluated. ENV735 grew slowly on MTBE or tert-butyl alcohol (TBA) as sole sources of carbon and energy, but growth on these substrates was greatly enhanced by the addition of a small amount of yeast extract. The addition of H(2) did not enhance or diminish MTBE degradation by the strain, and MTBE was only poorly degraded or not degraded by type strains of Hydrogenophaga or hydrogen-oxidizing enrichment cultures, respectively. MTBE degradation activity was constitutively expressed in ENV735 and was not greatly affected by formaldehyde, carbon monoxide, allyl thiourea, or acetylene. MTBE degradation was inhibited by 1-amino benzotriazole and butadiene monoepoxide. TBA degradation was inducible by TBA and was inhibited by formaldehyde at concentrations of >0.24 mM and by acetylene but not by the other inhibitors tested. These results demonstrate that separate, independently regulated genes encode MTBE and TBA metabolism in ENV735.


Formation of Type II Silicon Clathrate with Lithium Guests through Thermal Diffusion.
At low guest atom concentrations, Si clathrates can be viewed as semiconductors, with the guest atoms acting as dopants, potentially creating alternatives to diamond Si with exciting optoelectronic and spin properties. Studying Si clathrates with different guest atoms would not only provide insights into the electronic structure of the Si clathrates but also give insights into the unique properties that each guest can bring to the Si clathrate structure. However, the synthesis of Si clathrates with guests other than Na is challenging. In this study, we have developed an alternative approach, using thermal diffusion into type II Si clathrate with an extremely low Na concentration, to create Si clathrate with Li guests. Using time-of-flight secondary-ion mass spectroscopy, X-ray diffraction, and Raman scattering, thermal diffusion of Li into the nearly empty Si clathrate framework is detected and characterized as a function of the diffusion temperature and time. Interestingly, the Si clathrate exhibits reduced structural stability in the presence of Li, converting to polycrystalline or disordered phases for anneals at temperatures where the starting Na guest Si clathrate is quite stable. The Li atoms inserted into the Si clathrate lattice contribute free carriers, which can be detected in Raman scattering through their effect on the strength of Si-Si bonds in the framework. These carriers can also be observed in electron paramagnetic resonance (EPR). EPR shows, however, that Li guests are not simple analogues of Na guests. In particular, our results suggest that Li atoms, with their smaller size, tend to doubly occupy cages, forming "molecular-like" pairs with other Li or Na atoms. Results of this work provide a deeper insight into Li guest atoms in Si clathrate. These findings are also relevant to understanding how Li moves through and interacts with Si clathrate anodes in Li-ion batteries. Additionally, techniques presented in this work demonstrate a new method for filling the Si clathrate cages, enabling studies of a broad range of other guests in Si clathrates.


Study on Mechanical Properties and High-Speed Impact Resistance of Carbon Nanofibers/Polyurethane Composites Modified by Polydopamine.
Polyurethane elastomers (PUE), with superior mechanical properties and excellent corrosion resistance, are applied widely to the protective capability of structures under low-speed impact. However, they are prone to instantaneous phase transition, irreversible deformation and rupture even arising from holes under high-speed impact. In this paper, mussel adhesion proteins were applied to modify carbon nanofibers (CNFs) in a non-covalent way, and creatively mixed with PUE. This can improve the dispersity and interfacial compatibility of nanofillers in the PUE matrix. In addition, the homogeneous dispersion of modified nanofillers can serve as "reinforcing steel bars". The nanofillers and PUE matrix can form "mud and brick" structures, which show superb mechanical properties and impact resistance. Specifically, the reinforcement of 1.0 wt.% modified fillers in PUE is 103.51%, 95.12% and 119.85% higher than the neat PUE in compression modulus, storage modulus and energy absorption capability, respectively. The results have great implications in the design of composite parts for aerospace and army vehicles under extreme circumstances.


Effect of heat on synthesis of gelatinases and pro-inflammatory cytokines in equine tendinocytes.
The aim of this study was to clarify whether matrix metalloproteinases (MMP-2 and -9: gelatinases) and pro-inflammatory cytokines [tumor necrosis factor (TNF) alpha and interleukin (IL)-1beta] are induced by heat in tendon tissue in vitro and to test the hypothesis that heat exposure causes tendinocytes to synthesize pro-inflammatory cytokines and that synthesis of these cytokines, in turn, leads to up-regulation of synthesis of gelatinases. Isolated tendinocytes from equine superficial digital flexor tendons were cultured and all experiments were performed on cells passaged 3 or 4 times. In the cells exposed to heat (37 to 45 degrees C, 0 to 60 min), the survival rate decreased sharply in a temperature- and time-dependent manner, especially at 42 and 45 degrees C. Cells exposed at 40 degrees C, however, showed little change in survival rate and morphology. Gelatin zymograms revealed that proMMP-2 and -9 were the only two MMPs remaining in the supernatant of the cultured tendinocytes, including that of untreated cells. Addition of TNFalpha and IL-1beta to the culture medium of tendinocytes accelerated proMMP-9 synthesis considerably. Heating the tendinocytes (40 degrees C) led to a three-fold increase in proMMP-9 synthesis in a short time. Only TNFalpha was detected in tendinocytes after heat exposure for 30 and 60 min. In contrast, IL-1beta was under the detectable level in ELISA. Cooling of heat-exposed cells from 40 degrees C to 37 degrees C considerably down-regulated cellular proMMP-9 synthesis. Furthermore, proMMP-9 level was greatly reduced in cells treated at lower temperatures, 20 degrees C and 5 degrees C. These findings support our hypothesis that hyperthermia in the horse tendon induces tendinocytes to synthesize pro-inflammatory cytokines and that the synthesis of these cytokines results in the up-regulation of gelatinases.


Keratin immunoreactivity in the benign and neoplastic human prostate.
Keratin immunoreactivity in the benign and neoplastic human prostate was examined immunohistochemically using two monoclonal antibodies with differing specificities. One of these antibodies stained only the basal cells of the normal and hyperplastic prostatic epithelium, with no reactivity in tumor cells of prostatic adenocarcinoma. The other monoclonal antibody recognized a keratin protein present in all normal and hyperplastic columnar (secretory) epithelial cells, as well as in all cancer cells regardless of degree of tumor differentiation. In addition, the second antibody stained acinar and ductal epithelial cells exhibiting premalignant changes. Our findings indicate that keratin immunoreactivity differs among the epithelial cell populations of the human prostate, probably reflecting expression of different keratin proteins. The distinctive patterns of staining obtained with these two antibodies may assist in distinguishing hyperplastic from neoplastic prostatic epithelium, as well as in the recognition of basal cell hyperplasia, transitional cell metaplasia, and premalignant changes.


A population of in vivo anergized T cells with a lower activation threshold for the induction of CD25 exhibit differential requirements in mobilization of intracellular calcium and mitogen-activated protein kinase activation.
Chronic exposure of mature T cells with specificity for self-Ags can lead to the induction of a nonfunctional state which is referred to as T cell anergy. It is unclear whether anergic T cells are destined for cell death and thereby harmless or whether they can contribute to the induction of autoimmunity and/or regulation of anti-self reactivity. We have begun to address this issue. In a recent study, we showed that a population of mature CD4-CD8- T cells that express a transgenic TCR specific for the Ld MHC class I molecule are rendered anergic in Ld-expressing mice. In this study, we show that this population of anergic T cells possess a lower activation threshold for the induction of CD25 and CD69 in response to stimulation by antigenic ligands. Furthermore, these anergic T cells undergo extensive proliferation when stimulated with a low-affinity ligand in the presence of an exogenous source of IL-2. Biochemical analysis of the early intracellular signaling events of these in vivo anergized T cells showed that they have a signaling defect at the level of ZAP-70 and linker for the activation of T cell (LAT) phosphorylation. They also exhibit a defect in mobilization of intracellular calcium in response to TCR signaling. However, these anergic T cells demonstrate no defect in SLP-76 phosphorylation and extracellular signal-regulated kinase 1/2 activation. These biochemical characteristics of the anergic T cells were associated with an elevated level of Fyn, but not Lck expression. The potential contributions of these anergic T cells in the induction and/or regulation of autoimmune responses are discussed.


The associations of RENAL, PADUA and C-index nephrometry scores with perioperative outcomes and postoperative renal function in minimally invasive partial nephrectomy.
This study aimed to assess the utility of the radius, exophytic/endophytic, nearness, anterior/posterior, location (RENAL); preoperative aspects and dimensions used for an anatomic evaluation (PADUA), and centrality index (C-index) scores for the outcomes of partial nephrectomy (PN). The patients who underwent PN with contrast-enhanced preoperative imaging from January 2015 to June 2018 were identified. The RENAL, PADUA, and C-index scores were assigned. The correlation between these scoring systems and perioperative and long-term renal functional outcomes were evaluated. A total of 78 patients were included in the study (58 men and 20 women; age, 58±11.4 years). Median warm ischemia time (WIT), estimated blood loss (EBL), and operation time (OT) were 26 min, 115 mL, and 140 min, respectively. The RENAL score was related to WIT, EBL, and OT (p<0.001, p=0.003, and p=0.023, respectively). The PADUA score was associated with WIT, EBL, and OT (p<0.001, p=0.013, and p=0.005, respectively). The C-index score was correlated with WIT, EBL, and OT (p<0.001, p=0.010, and p=0.001, respectively). The C-index score also correlated with the percentage change in the estimated glomerular filtration rate (p=0.037). However, on univariable and multivariable regression analyses, only WIT significantly affected the postoperative estimated glomerular filtration rate reduction. The RENAL, PADUA, and C-index scores were significantly associated with perioperative outcomes of PN. In addition, the C-index score was correlated with long-term renal functional outcomes.


Multistep adhesion of Plasmodium sporozoites.
Adhesion of eukaryotic cells is a complex process during which interactions between extracellular ligands and cellular receptors on the plasma membrane modulate the organization of the cytoskeleton. Pathogens particularly rely often on adhesion to tissues or host cells in order to establish an infection. Here, we examined the adhesion of Plasmodium sporozoites, the motile form of the malaria parasite transmitted by the mosquito, to flat surfaces. Experiments using total internal reflection fluorescence microscopy and analysis of sporozoites under flow revealed a stepwise and developmentally regulated adhesion process. The sporozoite-specific transmembrane proteins TRAP and S6 were found to be important for initial adhesion. The structurally related protein TLP appears to play a specific role in adhesion under static conditions, as tlp(-) sporozoites move 4 times less efficiently than wild-type sporozoites. This likely reflects the decreased intradermal sporozoite movement of sporozoites lacking TLP. Further, these three sporozoite surface proteins also act in concert with actin filaments to organize efficient adhesion of the sporozoite prior to initiating motility and host cell invasion.


Pharmacokinetic/Pharmacodynamic Modeling To Predict the Antiplatelet Effect of the Ticagrelor-Loaded Self-Microemulsifying Drug Delivery System in Rats.
Ticagrelor (TCG) has been used as an antiplatelet agent for acute coronary syndrome patients. The aim of this research was to establish a population pharmacokinetic/pharmacodynamic (PK/PD) model of TCG and to apply the model for predicting the PD response of the TCG-loaded self-microemulsifying drug delivery system (TCG-SME) in rats. Pure TCG and TCG-SME (2, 5, and 10 mg/kg of TCG) were orally administered to male Sprague-Dawley rats. Plasma samples were collected at scheduled time-points and then analyzed for TCG plasma concentrations and antiplatelet effects. The inhibition of platelet aggregation of TCG was measured as a PD response. The PK profiles of pure TCG and TCG-SME could be well-explained with a two-compartment PK model. The accuracy of the PK model was assessed with a goodness-of-fit plot and conditional weight residual error (CWRES). Also, the visual predictive check was investigated based on the predictions. A population PK/PD model for pure TCG was established as an indirect response Emax model linked to the two-compartment PK model of pure TCG. The PK/PD model proposed a suitable fitting to link the plasma concentration of TCG simultaneously with platelet aggregation. Based on the PK data of TCG-SME, as well as the established PK/PD model of pure TCG, the PD profiles of TCG-SME were simulated. TCG-SME was more effective in inducing the antiplatelet effect than pure TCG at equivalent doses of TCG. The accuracy of the simulation was verified by comparing the simulated PD profile with the profile observed in rats. The observations were close to the model simulations. In addition, the values of CWRES were almost within ±2. In conclusion, the PK/PD modeling approach can provide a way for predicting mathematically the PD responses from PK profiles of other TCG formulations and a conceptual prediction for future clinical assessment.


Sample size calculations: basic principles and common pitfalls.
One of the most common requests that statisticians get from investigators are sample size calculations or sample size justifications. The sample size is the number of patients or other experimental units included in a study, and determining the sample size required to answer the research question is one of the first steps in designing a study. Although most statistical textbooks describe techniques for sample size calculation, it is often difficult for investigators to decide which method to use. There are many formulas available which can be applied for different types of data and study designs. However, all of these formulas should be used with caution since they are sensitive to errors, and small differences in selected parameters can lead to large differences in the sample size. In this paper, we discuss the basic principles of sample size calculations, the most common pitfalls and the reporting of these calculations.


Liver Transcriptome Profiling Reveals That Dietary DHA and EPA Levels Influence Suites of Genes Involved in Metabolism, Redox Homeostasis, and Immune Function in Atlantic Salmon (Salmo salar).
The optimal dietary requirement of omega-3 long-chain polyunsaturated fatty acids (ω3 LC-PUFA), namely docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), for Atlantic salmon that promotes growth and health warrants careful investigation. We used 44K microarrays to study the influence of increasing levels of dietary DHA + EPA (0, 1.0, and 1.4% of the diet, as formulated) in the presence of high linoleic acid (LA) on Atlantic salmon growth and liver transcriptome. After a 14-week feeding trial, Atlantic salmon fed diet ω3LC0 (i.e. 0% of DHA + EPA) showed significantly lower final weight and weight gain, and higher feed conversion ratio compared with ω3LC1.0 and ω3LC1.4 diet groups. The microarray experiment identified 55 and 77 differentially expressed probes (Rank Products analyses; PFP < 10%) in salmon fed diets ω3LC1.4 and ω3LC1.0 compared with those fed diet ω3LC0, respectively. The comparison between ω3LC1.4 and ω3LC1.0 revealed 134 differentially expressed probes. The microarray results were confirmed by qPCR analyses of 22 microarray-identified transcripts. Several key genes involved in fatty acid metabolism including LC-PUFA synthesis were upregulated in fish fed ω3LC0 compared with both other groups. Hierarchical clustering and linear regression analyses of liver qPCR and fatty acid composition data demonstrated significant correlations. In the current study, 1.0% ω3 LC-PUFA seemed to be the minimum requirement for Atlantic salmon based on growth performance; however, multivariate statistical analyses (PERMANOVA and SIMPER) showed that fish fed ω3LC1.0 and ω3LC1.4 diets had similar hepatic fatty acid profiles but marked differences in the transcript expression of biomarker genes involved in redox homeostasis (mgst1), immune responses (mxb, igmb, irf3, lect2a, srk2, and lyz2), and LC-PUFA synthesis (srebp1, fadsd5, and elovl2). This research has provided new insights into dietary requirement of DHA and EPA and their impact on physiologically important pathways in addition to lipid metabolism in Atlantic salmon.


The critical time window for androgen-dependent development of the Wolffian duct in the rat.
Androgens are thought to separately regulate stabilization and differentiation of the Wolffian duct (WD), but the time windows for these effects are unclear. To address this, fetal rats were exposed to flutamide within either an early window (EW) [embryonic day 15.5 (E15.5) to E17.5], when the WD degenerates in the female, or a later window (LW) (E19.5-E21.5), when the WD morphologically differentiates in the male, or during the full window of WD development (FW) (E15.5-21.5). WDs were examined for abnormalities during fetal (E21.5) or postnatal life, and anogenital distance and prostate presence/absence were recorded. Exposure to FW- or EW-flutamide, but not to LW-flutamide, induced comparable abnormalities in the fetal WD at E21.5, namely reduced WD coiling, reduced cell proliferation, reduced epithelial cell height, altered epithelial vimentin expression, and reduced expression of smooth muscle actin in the WD inner stroma. Exposure to EW- or FW-flutamide, but not to LW-flutamide, resulted in incomplete/absent WDs in more than 50% of males by adulthood, although such abnormalities were infrequent in fetal life. These findings suggest that androgen action during the EW is sufficient to promote WD morphological differentiation several days later. Because the androgen receptor is expressed in the WD stroma but not in the epithelium during this EW, WD differentiation is likely to be dependent on androgen-mediated signaling from the stroma to the epithelium. In conclusion, the critical window for androgen action in regulating WD development in the rat is between E15.5 and E17.5. This window is also important for prostate formation and anogenital distance masculinization.


Towards defining the core Saccharum microbiome: input from five genotypes.
Plant microbiome and its manipulation inaugurate a new era for plant biotechnology with the potential to benefit sustainable crop production. Here, we used the large-scale 16S rDNA sequencing analysis to unravel the dynamic, structure, and composition of exophytic and endophytic microbial communities in two hybrid commercial cultivars of sugarcane (R570 and SP80-3280), two cultivated genotypes (Saccharum officinarum and Saccharum barberi) and one wild species (Saccharum spontaneum). Our analysis identified 1372 amplicon sequence variants (ASVs). The microbial communities' profiles are grouped by two, root and bulk soils and stem and leave when these four components are compared. However, PCoA-based data supports that endophytes and epiphytes communities form distinct groups, revealing an active host-derived mechanism to select the resident microbiota. A strong genotype-influence on the assembly of microbial communities in Saccharum ssp. is documented. A total of 220 ASVs persisted across plant cultivars and species. The ubiquitous bacteria are two potential beneficial bacteria, Acinetobacter ssp., and Serratia symbiotica. The results presented support the existence of common and cultivar-specific ASVs in two commercial hybrids, two cultivated canes and one species of Saccharum across tissues (leaves, stems, and roots). Also, evidence is provided that under the experimental conditions described here, each genotype bears its microbial community with little impact from the soil conditions, except in the root system. It remains to be demonstrated which aspect, genotype, environment or both, has the most significant impact on the microbial selection in sugarcane fields.


"Humic coverage index" as a determining factor governing strain-specific hydrocarbon availability to contaminant-degrading bacteria in soils.
We report development of a novel parameter for quantifying the amount of humic and fulvic acids per unit surface area in a particular soil. This quantity, the "humic coverage index" (HCI), provides a measurement of the relative spatial extents and/or thicknesses of the humic/fulvic overlayers in different soils, and, therefore, can be used in modeling various soils' behavior in sequestration processes in which humic materials are involved. HCI is herein applied to modeling biodegradation of aromatic and aliphatic hydrocarbons (phenanthrene, pyrene, and hexadecane) by several bacterial strains. Results indicate that, for the cases studies here, contaminant biodegradation is highest at a particular HCI and decreases if the coverage density of humic material is lower or higher than this optimum value. The HCI value at which maximal degradation was observed varied across different strains (indicating strain-specific differences in ability to degrade contaminants sorbed to humic materials) and, to a lesser extent, across different contaminants. The HCI concept is also demonstrated to be useful in explaining soil-, strain-, and contaminant-specific variations in the ability of fulvic acid supplementation to enhance contaminant biodegradation. Finally, we show that, in general, strains which are comparatively better at degrading contaminants in high-HCI soils also show enhanced contaminant mineralization in vitro in the presence of humic acids, such as when hydrocarbons are adsorbed onto these materials.


Glycosylation patterns and PHA-E-associated glycoprotein profiling associated with early hepatic encephalopathy in Chinese hepatocellular carcinoma patients.
Hepatic encephalopathy (HE) as a severe neuropsychiatric complication is commonly present in the end stage of Hepatocellular Carcinoma (HCC). However, widely accepted biomarkers for diagnosing early HE are still absent. Here, we screened glycosylation patterns of serum proteins from Chinese HCC patients with or without early HE by lectin microarray. Then, phaseolus vulgaris erythroagglutinin (PHA-E) as a lectin binding with bisecting GlcNAc structure which was significantly decreased in sera from Chinese HCC patients with early HE, was chosen to perform lectin affinity chromatography, following by in-gel digestion, Mass Spectrometry (MS) analysis and bioinformatics analysis. Here we found, 13 lectins showed statistically significant reduction suggesting GalNAc, terminal α-1,3 Man, bisecting GlcNAc, (GlcNAc)n, O-GlcNAc, Neu5Ac, tetra-antennary complex-type N-glycan and GalNAc α/β1-3/6 Gal were decreased in serum glycoproteins from Chinese HCC patients with early HE. Furthermore, a total of 141 PHA-E-associated glycoproteins were identified in MS, of which 12 serum glycoproteins only in Chinese HCC patients without early HE and 26 serum glycoproteins only in Chinese HCC patients with early HE. In addition, bioinformatics analysis revealed the PHA-E-associated serum glycoproteins only in Chinese HCC patients with early HE might be related to early HE occurrence through p38 MAPK signaling pathway and MAPK/ERK signaling pathway. Collectively, this was the first glycomics study of serum proteins in HCC patients with early HE and it could provide a database for discovering and developing serum biomarkers to identify and predict early HE in HCC patients.


Statistical and conformational analysis of the electron density of protein side chains.
Protein side chains make most of the specific contacts between proteins and other molecules, and their conformational properties have been studied for many years. These properties have been analyzed primarily in the form of rotamer libraries, which cluster the observed conformations into groups and provide frequencies and average dihedral angles for these groups. In recent years, these libraries have improved with higher resolution structures and using various criteria such as high thermal factors to eliminate side chains that may be misplaced within the crystallographic model coordinates. Many of these side chains have highly non-rotameric dihedral angles. The origin of side chains with high B-factors and/or with non-rotameric dihedral angles is of interest in the determination of protein structures and in assessing the prediction of side chain conformations. In this paper, using a statistical analysis of the electron density of a large set of proteins, it is shown that: (1) most non-rotameric side chains have low electron density compared to rotameric side chains; (2) up to 15% of chi1 non-rotameric side chains in PDB models can clearly be fit to density at a single rotameric conformation and in some cases multiple rotameric conformations; (3) a further 47% of non-rotameric side chains have highly dispersed electron density, indicating potentially interconverting rotameric conformations; (4) the entropy of these side chains is close to that of side chains annotated as having more than one chi(1) rotamer in the crystallographic model; (5) many rotameric side chains with high entropy clearly show multiple conformations that are not annotated in the crystallographic model. These results indicate that modeling of side chains alternating between rotamers in the electron density is important and needs further improvement, both in structure determination and in structure prediction.


Distribution of remoxipride to the human brain and central D2-dopamine receptor binding examined in vivo by PET.
Remoxipride is a new antipsychotic drug that binds selectively to the D2-dopamine receptor subtype as demonstrated in animal studies in vitro and in vivo. It is generally assumed that the antipsychotic effect of neuroleptic drugs is mediated by blockade of dopamine receptors. The aim of the present study was to use positron emission tomography (PET) and the ligand [11C] raclopride to examine the central D2-dopamine receptor occupancy in man during oral administration of remoxipride. After oral administration of remoxipride 100 mg three times daily to a healthy male subject there was a 73% central D2-dopamine receptor occupancy. In a schizophrenic patient treated with remoxipride 200 mg twice daily there was a 71% occupancy. These occupancy values are similar to the values of 65-85% previously found in a series of patients treated with neuroleptics representative of all currently used chemical classes. In a separate experiment, remoxipride was labelled with 11C and injected intravenously and the distribution of radioactivity to the brain examined. Radioactivity appeared in the brain during the first minutes after injection and 4.5 min after injection it accounted for 0.8% of the total radioactivity injected, thus indicating that [11C]remoxipride had rapidly passed through the blood-brain barrier.


Immunoreactivity to neurofilaments in the rodent anterior pituitary is associated with the expression of alpha 1A protein subunits of voltage-gated Ca2+ channels.
We recently reported that rodent anterior pituitary (AP) cells (with the exception of corticotrophs and melanotrophs) express neuronal markers, including 68-kDa neurofilaments (NF68) in an oestrogen-dependent manner. The functional significance of neurofilament (NF) expression in the AP is unknown, but recent data in myelinated nerve fibres from NF-null mice suggest that NFs can regulate ion channel function. Because Ca(2+) influx through voltage-gated Ca(2+) channels is required for hormone secretion in AP cells, and oestrogen regulates the expression of Ca(2+) channels in AP cells, the present study examined the expression of alpha1 subunits of voltage gated Ca(2+) channels in relation to that of NF68. Using quantitative immunofluorescence, we demonstrate that alpha 1C and alpha 1D subunits are abundantly expressed in female AP cells, alpha 1A subunits are moderately expressed, and alpha 1G and alpha 1B subunits are expressed at the lowest levels. Double-immunostaining showed that NF68 expression is not correlated with that of alpha 1C, alpha 1D or alpha 1B. Expression of alpha 1G and NF68 appear to be mutually exclusive from each other. Moreover, alpha 1A subunit and NF68 expression are significantly correlated and alpha 1A immunoreactivity is sexually dimorphic (i.e. low in males and high in females) and its levels of expression vary during the oestrous cycle, similar to NF68. Finally, omega-agatoxin IVA, a specific blocker of P/Q type Ca(2+) currents that are a result of the activity of alpha 1A subunits, inhibited to a greater extent spontaneous [Ca(2+)](i) fluctuations in AP cells from females in oestrous and dioestrous, whereas cells from females in pro-oestrous and males were less affected by this toxin. These results suggest a preferential participation of P/Q-type Ca(2+) channels and hence alpha 1A subunits, in regulating spontaneous Ca(2+) transients in AP cells under conditions where the proportion of NF68-expressing cells is high. It remains to be determined whether the expression of NF68 affects that of alpha 1A Ca(2+) channel subunits or vice versa.


The response of newborn sheep to TRH with and without somatostatin.
Hormone responses to a bolus injection of thyrotropin releasing hormone (TRH) were studied in 8 newborn lambs between 6 and 19 hours of age. The effect of a bolus injection and 45 min infusion of somatostatin (SRIF) on these responses was studied in 2 other animals. Serial measurements of serum TSH, prolactin, triiodothyronine (T3) and thyroxine (T4) were conducted for 2 to 6 h in all animals. Mean baseline T4 and T3 concentrations were 12.6 microng/dl and 221 ng/dl, respectively, both significantly higher than values in fetal or adult animals. These high values were due to the events of parturition. In spite of the high baseline T4 and T3 levels, there were rapid and significant increases in both TSH and prolactin concentrations in response to TRH alone. The TSH response evoked further increments in serum T3 and T4 concentrations observed at 30 min and 60 min, respectively, both subsequently increasing progressively through 6 h. During the 45 min period of SRIF infusion, the TSH T4 and T3 responses to the zero time TRH injection were minimal. However, after discontinuing SRIF, late increases in TSH, T4 and T3 were observed. The results indicate that the hyperiodothyroninemia characteristic of the newborn period does not block the response to exogenous TRH, whereas the inhibitory effect of exogenous SRIF is observed in the newborn as in the adult. The increased endogenous TRH secretion presumably responsible for the neonatal TSH surge may be overriding the negative feedback effect of thyroid hormones.


Towards a better QALY model.
This paper presents a test of the predictive validity of various classes of QALY models (i.e. linear, power and exponential models). We first estimated TTO utilities for 43 EQ-5D chronic health states and next these states were embedded in nonchronic health profiles. The chronic TTO utilities were then used to predict the responses to TTO questions with nonchronic health profiles. We find that the power QALY model clearly outperforms linear and exponential QALY models. Optimal power coefficient is 0.65. Our results suggest that TTO-based QALY calculations may be biased. This bias can be corrected using a power QALY model.


The limits of mean-field heterozygosity estimates under spatial extension in simulated plant populations.
Computational models of evolutionary processes are increasingly required to incorporate multiple and diverse sources of data. A popular feature to include in population genetics models is spatial extension, which reflects more accurately natural populations than does a mean field approach. However, such models necessarily violate the mean field assumptions of classical population genetics, as do natural populations in the real world. Recently, it has been questioned whether classical approaches are truly applicable to the real world. Individual based models (IBM) are a powerful and versatile approach to achieve integration in models. In this study an IBM was used to examine how populations of plants deviate from classical expectations under spatial extension. Populations of plants that used three different mating strategies were placed in a range of arena sizes giving crowded to sparse occupation densities. Using a measure of population density, the pollen communication distance (P(cd)), the deviation exhibited by outbreeding populations differed from classical mean field expectations by less than 5% when P(cd) was less than 1, and over this threshold value the deviation significantly increased. Populations with an intermediate mating strategy did not have such a threshold and deviated directly with increasing isolation between individuals. Populations with a selfing strategy were influenced more by the mating strategy than by increased isolation. In all cases pollen dispersal was more influential than seed dispersal. The IBM model showed that mean field calculations can be reasonably applied to natural outbreeding plant populations that occur at a density in which individuals are less than the average pollen dispersal distance from their neighbors.


Optimization of regional emergency supplies distribution vehicle route with dynamic real-time demand.
Given the particular characteristics of a sudden outbreak of an epidemic on a regional scale and considering the possible existence of a latent period process, this paper takes the distribution of regional emergency supplies as the research object. Form the proposes a dynamic vehicle path problem from the perspective of real-time demand changes. First, when there is a sudden outbreak of a small-scale epidemic, there is uncertainty about demand in the epidemic area. The objective functions of minimizing the vehicle travel route cost of emergency vehicles, the late arrival penalty cost of emergency vehicles, and the fixed cost of emergency vehicles, as well as the objective function of minimizing the total distance traveled by vehicles, are established. Second, a mathematical model of the dynamic real-time demand vehicle route problem is built using the actual vehicle routing problem as a basis. The model is then solved using the SFSSA method. Finally, the computational results demonstrate that the SFSSA algorithm can effectively reduce transportation cost and distance when solving the constructed mathematical model problem, providing a solution to the problem of optimizing the route of emergency material distribution vehicles for a regional scale.