Sublethal effects on wood frogs chronically exposed to environmentally relevant concentrations of two neonicotinoid insecticides.
Neonicotinoids are prophylactically used globally on a variety of crops, and there is concern for the potential impacts of neonicotinoids on aquatic ecosystems. The intensive use of pesticides on crops has been identified as a contributor to population declines of amphibians, but currently little is known regarding the sublethal effects of chronic neonicotinoid exposure on amphibians. The objective of the present study was to characterize the sublethal effect(s) of exposure to 3 environmentally relevant concentrations (1 μg/L, 10 μg/L, and 100 μg/L) of 2 neonicotinoids on larval wood frogs (Lithobates sylvaticus) using outdoor mesocosms. We exposed tadpoles to solutions of 2 commercial formulations containing imidacloprid and thiamethoxam, and assessed survival, growth, and development. Exposure to imidacloprid at 10 μg/L and 100 μg/L increased survival and delayed completion of metamorphosis compared with controls. Exposure to thiamethoxam did not influence amphibian responses. There was no significant effect of any treatment on body mass or size of the metamorphs. The results suggest that current usage of imidacloprid and thiamethoxam does not pose a threat to wood frogs. However, further assessment of both direct and indirect effects on subtle sublethal endpoints, and the influence of multiple interacting stressors at various life stages, is needed to fully understand the effects of neonicotinoids on amphibians. Environ Toxicol Chem 2017;36:1101-1109. © 2017 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.


Characterization of Pepper leafroll chlorosis virus, a New Polerovirus Causing Yellowing Disease of Bell Pepper in Saudi Arabia.
During the growing seasons of 2014 through 2016, a total of 336 leaf samples from bell pepper (showing leafroll and interveinal yellowing) and arable weeds were collected from Riyadh region, Saudi Arabia. The use of a polerovirus generic reverse transcription (RT)-PCR assay confirmed their presence in the bell pepper samples. Sequencing of the generic amplicon revealed high similarity (87.6 to 98.1% in nt) with four poleroviruses; Tobacco vein distorting virus, Pepper vein yellows virus, Pepper yellows virus, and Pepper yellow leaf curl virus. To further characterize one of these isolates (105D), a larger part of the genome (∼1,300 nt) spanning approximately from the 3' end of ORF2 to the middle of ORF3, was amplified and sequenced. Blasting the resulting sequence revealed the low amino acid and nucleotide identity percentages in the coat protein and movement protein partial genes with viruses deposited in GenBank. Next-generation sequence was used to acquire a larger part of the genome, which resulted in the reconstruction of isolate 105D's partial genome (5,496 nt). Sequence similarity analysis revealed the presence of a divergent polerovirus isolate belonging to a new species that was tentatively named Pepper leafroll chlorosis virus (PeLRCV). Using a specific RT-PCR assay for this isolate confirmed the presence of this new viral species in the symptomatic peppers. Aphid transmission experiments showed that PeLRCV is vectored by Aphis gossypii and that it can infect at least five out of the 15 different plants species tested. Based on our findings, PeLRCV is a new member of genus Polerovirus in the family Luteoviridae.


High-dose-rate interstitial brachytherapy in head and neck cancer: do we need a look back into a forgotten art - a single institute experience.
To evaluate the treatment outcomes with high-dose-rate (HDR) interstitial brachytherapy (HDR-BRT) in head and neck cancers (HNC). Fifty-eight patients with HNC as per American Joint Committee on Cancer (AJCC) TNM staging criteria were analyzed retrospectively between 2008 and 2015. Forty-two patients received external beam radiotherapy (EBRT) with HDR-BRT and 16 patients received BRT alone. The survival was calculated with respect to median biological equivalent doses (BED) and median 2 Gy equivalent dose (EQD2), keeping α/β = 10 for tumor. Loco-regional control and disease free survival was assessed. The median follow-up period was 25 months (2-84 months). The disease-free survival (DFS) probability at year 1 was 82.7%, and 68% at year 7. The overall survival probability was 91.3% at year 1 and 85.8% at year 7. The local control rate was 70%. The rate of recurrence was 30%. Distant metastasis rate was 17.2%. The median BED and EQD2, respectively, were 86.78 Gy and 71.6 Gy. The DFS was 74.1% and 75.9% in patients receiving a dose more than median BED and EQD2, respectively, and was 64.8% and 61.5% for less than the median dose. The overall outcome was good with implementation of HDR-BRT used alone or as boost, and shows DFS as better when the dose received is more than the median BED and median EQD2. The role of HDR-BRT in HNC is a proven, effective, and safe treatment method with excellent long term outcome as seen in this study, which reflects the need for reviving the forgotten art and science of interstitial brachytherapy in HNC.


ADP-induced platelet shape change and mobilization of cytoplasmic ionized calcium are mediated by distinct binding sites on platelets: 5'-p-fluorosulfonylbenzoyladenosine is a weak platelet agonist.
Platelet stimulation with ADP results in several responses, including shape change, increase in cytoplasmic ionized calcium concentration [Ca2+]i, an inhibition of adenylate cyclase. 5'-p-Fluorosulphonyl benzoyladenosine (FSBA), which covalently labels an ADP binding site on platelets, blocks platelet shape change but not the inhibition of cyclic AMP levels by ADP, whereas p-chloromercuribenzenesulfonate (pCMBS), a nonpenetrating thiol reagent, has the opposite effects. We examined the effect of FSBA and pCMBS on ADP-induced increase in [Ca2+]i using platelets loaded with fluorescent Ca2+ indicators quin2 and fura-2. FSBA (50 to 200 mumol/L) induced a dose-dependent rise in [Ca2+]i, indicating that it is a weak platelet agonist. Under conditions of covalent labeling of the ADP binding sites, FSBA (50 to 100 mumol/L) did not inhibit the ADP-induced increase in [Ca2+]i or its inhibition of adenylate cyclase, whereas pCMBS (up to 1 mmol/L) abolished both these responses but not shape change. These findings suggest that ADP-induced Ca2+ mobilization and inhibition of adenylate cyclase are mediated by platelet binding sites distinct from those mediating shape change.


Evaluation of the safety profiles of estrogenic Chinese herbal medicines in breast cancer.
An increasing number of breast cancer patients in Asian countries has been found to consume dietary supplements including phytoestrogen-rich Chinese herbal medicines with an expectation to alleviate the side effects of conventional cancer therapies. The question of whether estrogenic Chinese herbal medicines are beneficial or detrimental to the health of breast cancer patients remains uncertain. The present study aimed at establishing a systematic approach to look at the safety profiles of estrogenic Chinese herbal medicines (CHM). The effects of estrogenic CHM on the growth of human breast cancer cells as well as the progression of breast tumors in mice have been investigated. Our results demonstrated that among 10 selected estrogenic CHM, the aqueous extracts of Cistanche deserticola (CD) and Dioscorea opposita (DO) at 0.4 to 1.6 mg/ml significantly stimulated cell viability in both estrogen receptor (ER)-positive (MDA-MB-361 and MCF-7) and ER-negative (SKBR3 and MDA-MB-231) breast cancer cells. However, results from animal studies showed that no significant difference was found on the size of mouse 4T1 breast tumors in CD- and DO-treated mice when compared with the control group, while the number of proliferative cells were found to be increased in DO-treated group. Besides, CD and DO treatments induced significant immunomodulatory effects on 4T1 tumor-bearing mice by increasing the production of cytokines IL-2 and IFN-γ and modulation of regulatory T-cells. Furthermore, CD and DO treatments did not stimulate, but in fact suppressed human triple-negative MDA-MB-231 breast xenografts growth in immunodeficiency mice. The considerable concerns on the use of CD and DO in breast cancer patients could be relieved to some extents upon the findings of this pre-clinical study. The potential harmful effects of estrogenic Chinese herbal medicines on breast cancer growth should be verified in both cell-based and tumor-bearing mice models.


Silencing CAPN2 Expression Inhibited Castration-Resistant Prostate Cancer Cells Proliferation and Invasion via AKT/mTOR Signal Pathway.
The mRNA expression of CAPN2 was upregulated in CRPC cells (DU145 and PC3) than that in non-CRPC cells. Silencing CAPN2 expression could inhibit DU145 and PC3 cells proliferation by cell cycle arrest at G1 phase. Knockdown of CPAN2 level suppressed the migration and invasion capacity of CRPC cells by reducing matrix metalloproteinase-2 (MMP-2) and MMP-9 activation, as well as repressing the phosphorylation protein expression of AKT and mTOR. In addition, we found that the expression of CAPN2 was elevated in Pca tissues than that in normal control tissues. Therefore, we showed the important roles of CAPN2 in the development and progression in CRPC cells, suggesting a new therapeutic intervention for treating castration-resistant prostate cancer patients.


In vitro control of plant pathogenic Xanthomonas spp. using Poncirus trifoliata Rafin.
The secondary metabolites such as essential oil and pure compounds (limonin and imperatorin) from Poncirus trifoliata Rafin were tested for in vitro control of phytopathogenic bacteria of Xanthomonas spp. In vitro studies showed that the oil had inhibitory effect on Xanthomonas campestris pv. compestris KC94-17-XCC, Xanthomonas campestris pv. vesicatoria YK93-4-XCV, Xanthomonas oryzae pv. oryzae KX019-XCO and Xanthomonas sp. SK12 with their inhibition zones and minimum inhibitory concentration (MIC) values ranging from 13.1~22.1 mm and 62.5~125 μg/ml, respectively. Limonin and imperatorin also had in vitro antibacterial potential (MIC: 15.62~62.5 μg/ml) against all the tested Xanthomonas spp. Furthermore, the SEM studies demonstrated that limonin and imperatorin caused morphological changes of Xanthomonas sp. SK12 at the minimum inhibitory concentration (15.62 μg/ml). These results of this study support the possible use of essential oil and natural compounds from P. Trifoliata in agriculture and agro-industries to control plant pathogenic microorganisms.


Tobacco dependence among people with mental illness: a facility-based cross sectional study from Southwest Ethiopia.
Tobacco smoking is a health care issue in developed as well as in developing countries. Tobacco smoking among people with mental illness is significantly higher than in the general population. Tobacco smoking has negative effects on physical, mental and financial well-being of people with mental illness but little is known about tobacco dependence among mental health service users in sub-Saharan African countries, including Ethiopia. Therefore, this study attempted to assess the prevalence of tobacco dependence and associated factors among mental health service users at Jimma University teaching hospital. A cross-sectional study was conducted among 305 male and 117 female mental health service users at Jimma University teaching Hospital in 2014. The Fagerstrom Test for Nicotine Dependence (FTND) was used to assess tobacco dependence. Logistic regression analysis was used for bivariate and multivariate analysis. Variables with a P value of <0.05 were considered to be associated with tobacco dependence in the final model. The prevalence of current tobacco dependence among the study participants was 18.5%. Amongst people with tobacco dependence, 57.7, 29.5 and 12.8% had moderate, high and very high level of tobacco dependence respectively. All mental health service users with tobacco dependence were males. There was a significant association between tobacco dependence and high school education (AOR 3.02, 95% CI 1.07, 8. 48), alcohol use disorder (AOR 4.14, 95% CI = 1.54, 11.11), daily khat chewing (AOR 13.51, 95% CI = 4.27, 42.74), chewing khat 2-3 times per week (AOR 5.09, 95% CI = 1.37,18.95), chewing khat once a week (AOR 4.31, 95%CI = 1.04,17.78), having friends who smoke tobacco (AOR 4.88, 95% CI = 2.12, 11.25) and being diagnosed with schizophrenia compared to a diagnosis of major depression (AOR 5.26, 95% CI = 2.07, 13.37). However, daily attendance at a place of worship (AOR 0.46, 95% CI = 0.22, 0.95) was associated negatively with tobacco dependence. In this study, there was a high prevalence of tobacco dependence among mental health services users. There is a pressing need to increase availability of psychological and pharmacological interventions to reduce tobacco dependence and tobacco-related medical illness in this vulnerable group.


A near haploid clone: 24,XY, t (9; 22) (q34; q11) from a patient in blast crisis of chronic myeloid leukaemia.
Chromosome studies on bone marrow in a patient with chronic myeloid leukaemia (CML) revealed a mosaic picture with a dominant clone of 24,XY, t(9; 22) (q34; q11). This corresponded to a preponderance of minute blasts in smears of bone marrow aspirate. It is suggested that a haploidy event rather than progressive hypodiploid loss, is responsible for the genesis of the miniature blast cells.


Overexpression of MYB115, AAD2, or AAD3 in Arabidopsis thaliana seeds yields contrasting omega-7 contents.
Omega-7 monoenoic fatty acids (ω-7 FAs) are increasingly exploited both for their positive effects on health and for their industrial potential. Some plant species produce fruits or seeds with high amounts of ω-7 FAs. However, the low yields and poor agronomic properties of these plants preclude their commercial use. As an alternative, the metabolic engineering of oilseed crops for sustainable ω-7 FA production has been proposed. Two palmitoyl-ACP desaturases (PADs) catalyzing ω-7 FA biosynthesis were recently identified and characterized in Arabidopsis thaliana, together with MYB115 and MYB118, two transcription factors that positively control the expression of the corresponding PAD genes. In the present research, we examine the biotechnological potential of these new actors of ω-7 metabolism for the metabolic engineering of plant-based production of ω-7 FAs. We placed the PAD and MYB115 coding sequences under the control of a promoter strongly induced in seeds and evaluated these different constructs in A. thaliana. Seeds were obtained that exhibit ω-7 FA contents ranging from 10 to >50% of the total FAs, and these major compositional changes have no detrimental effect on seed germination.


Stimulation of poliovirus proteinase 3Cpro-related proteolysis by the genome-linked protein VPg and its precursor 3AB.
Purified recombinant poliovirus polypeptide 3AB interacts with 3CDpro and 3Dpol as shown by coimmunoprecipitation with anti-3Dpol antibodies. A consequence of this interaction is an accelerated autoprocessing of 3CDpro to produce 3Cpro and 3Dpol. The activation of 3Dpol polymerase activity by cleavage of 3CDpro, a polypeptide that has no polymerase activity, can be shown by template- and primer-dependent poly(U) synthesis. Anti-VPg antibodies (VPg = 3B) added to HeLa translation extracts programmed with poliovirion RNA inhibit cleavage of 3CDpro whereas addition of purified 3AB or VPg to these translation reactions increases 3CDpro processing. 3AB stimulates also 3Cpro-related proteolysis of 2BC, a poliovirus-specific, nonstructural processing intermediate. In contrast, 3CDpro-specific cleavage of the structural precursor P1 is inhibited by the addition of 3AB as shown by a decrease in the production of VP0 and VP3. These data shed new light on a phenomenon in the regulation of expression of poliovirus genetic information: whereas the proteinase 3CDpro is needed for processing of the capsid precursor, the cleavage product of this relatively stable precursor is required for RNA replication.


CD44 Assists the Topical Anti-Psoriatic Efficacy of Curcumin-Loaded Hyaluronan-Modified Ethosomes: A New Strategy for Clustering Drug in Inflammatory Skin.
Background: Psoriasis is a common chronic inflammatory skin disease. Its treatment is challenged by the limited amount of drug reaching the inflamed skin. The overexpressed CD44 protein in inflamed psoriatic skin can serve as a potential target of novel active-targeting nanocarriers to increase drug accumulation in the skin. Methods: Hyaluronic acid (HA) was linked to propylene glycol-based ethosomes by covalent binding to develop a novel topical drug delivery carrier (HA-ES) for curcumin. An imiquimod-induced psoriasis mouse model was established, and curcumin delivery and anti-psoriatic efficacy using HA-ES were compared with those using plain ethosomes (ES). Results: The HA gel network formed on the surface of HA-ES reduced the leakage and release of poorly water-soluble curcumin. Compared with ES, transdermal curcumin delivery was significantly enhanced by using HA-ES as vehicles; the cumulative transdermal amount and the amount retained in the skin in vitro after 8 h were, respectively, 1.6 and 1.4 times those observed with ES, as well as 3.1 and 3.3 times those observed with a curcumin propylene glycol solution (PGS), respectively. The in vivo psoriatic skin retention of curcumin with HA-ES was 2.3 and 4.0 times that of ES and PGS, respectively. CD44 expression in imiquimod-induced psoriasis-like inflamed skin was 2.7 times that in normal skin. Immunostaining revealed similar results, suggesting that the specific adhesion of HA-ES to CD44 increased drug accumulation in the skin. After topical administration to mice, the HA-ES group showed an alleviation of inflammation symptoms; lower TNF-α, IL-17A, IL-17F, IL-22, and IL-1β mRNA levels; and lower CCR6 protein expression compared to the ES and PGS groups. Conclusion: We demonstrated increased topical drug delivery of curcumin to inflamed tissues using HA-ES targeting the highly expressed CD44 protein. This innovative strategy could be applied for the development of topical drug delivery systems targeting inflamed skin.


[Salivary peptide P-C (III). Its relation to the pathogenesis of diabetes mellitus].
Previous immunohistochemical study showed that salivary peptide P-C like immunoreactivity, originally isolated from whole human saliva, was present not only in human salivary glands but also in human pancreatic B-cells. To elucidate the pathophysiological role of this peptide-like immunoreactivity in human pancreatic B-cells, immunohistochemical study using antisera against both insulin and peptide P-C was carried out on the paraffin embedded pancreatic tissues of 27 diabetic patients and 30 control subjects. Positive immunofluorescence due to insulin was detected in 96% of the diabetic pancreases and 100% of the controls. A pancreas of only one IDDM did not have any immunofluorescence due to insulin. Thus, no significant differences were seen in connection with the presence of insulin between pancreases of NIDDM and the controls, though the present study did not examine whether the number of pancreatic B-cells in the pancreas of NIDDM was the same as that in the controls or not. In contrast, positive immunofluorescence due to peptide P-C like immunoreactivity was demonstrated in only 41% of the diabetic pancreases but in 87% of the controls. In view of the fact that the diabetic pancreases and the controls were fixed and embedded into paraffin in the same department of pathology and under the same conditions, the negative finding for salivary peptide P-C like immunoreactive in the diabetic pancreas seems to result neither from the effects of fixatives nor the destructive postmortem effects of enzymes on the antigenicity of peptide P-C like immunoreactivity but from the decreased content of peptide P-C like immunoreactivity in the diabetic pancreas.(ABSTRACT TRUNCATED AT 250 WORDS)


[Neuropathology and immunohistochemistry of brains with Rett syndrome].
To clarify critical lesions and pathomechanism of Rett syndrome (RTT), we studied the followings: (1) neuropathological observation of RTT, (2) Comparison of neurotransmitter expression between RTT and controls, and developmental immunohistology of MeCP2, the product of the causative gene of RTT in control brains using the ABC immunohistochemical technique. In the RTT brainstem, there were lesions in the substantia nigra, periaquaductal gray and locus ceruleus, in which catecholaminergic neurons were markedly reduced. MeCP2 expression, was detectable in all neurons of fetal brain until 20 gestational weeks (GW). The expression disappeared in the cerebrum after 20 GW and in the brainstem after perinatal or infantile periods, but re-appeared in the brainstem after adolescence. These findings suggest that the period and location of MeCP2 expression may play an important role in the pathogenesis of RTT. The lack of MeCP2 in the specific periods and regions may lead to the dysfunction of catecholaminergic neurons in RTT brains.


The prevention of musculoskeletal complaints: a randomized controlled trial on additional effects of a work-related psychosocial coaching intervention compared to physiotherapy alone.
Research shows that psychosocial factors play a significant role on the emergence of musculoskeletal complaints (MSC). The aim of this study was to determine whether a coaching intervention which was focussed on enabling better strategies for coping with work stressors is superior to physiotherapy alone in the reduction of MSC. 68 nurses were randomized to an intervention group (IG, n = 34) or a control group (CG, n = 34). The IG and CG completed a weekly individual physiotherapy unit (10 weeks). Additionally, the IG passed five coaching sessions (fortnightly), plus one opening and one closing session. The primary outcome was MSC, secondary outcomes were work ability and work-related wellbeing. Outcomes were obtained by physical examinations and questionnaires. Data were analyzed by t-test, Chi-Square test, ANOVA with repeated measurements, and multilevel analyzes. In respect of MSC, the IG compared to the CG showed a significant improvement in the pain severity of everyday movements, and trends towards an improvement of movement in the vertebral column as well as a reduction of the pain severity due to maximum degree movements. No effects were observed in respect to muscle strengths, and restrictions of everyday activities. The IG exhibited a significant improvement of work ability in reference to the physical working demands, and work-related wellbeing. Analysis indicates that improvements in the IG increased further in the 12 weeks after the intervention. The results suggest that the coaching, beyond physiotherapy, can support the reduction of MSC, the improvement of work ability and work-related wellbeing.


Metagenomic and metatranscriptomic analysis of microbial community structure and gene expression of activated sludge.
The present study applied both metagenomic and metatranscriptomic approaches to characterize microbial structure and gene expression of an activated sludge community from a municipal wastewater treatment plant in Hong Kong. DNA and cDNA were sequenced by Illumina Hi-seq2000 at a depth of 2.4 Gbp. Taxonomic analysis by MG-RAST showed bacteria were dominant in both DNA and cDNA datasets. The taxonomic profile obtained by BLAST against SILVA SSUref database and annotation by MEGAN showed that activated sludge was dominated by Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes and Verrucomicrobia phyla in both DNA and cDNA datasets. Global gene expression annotation based on KEGG metabolism pathway displayed slight disagreement between the DNA and cDNA datasets. Further gene expression annotation focusing on nitrogen removal revealed that denitrification-related genes sequences dominated in both DNA and cDNA datasets, while nitrifying genes were also expressed in relative high levels. Specially, ammonia monooxygenase and hydroxylamine oxidase demonstrated the high cDNA/DNA ratios in the present study, indicating strong nitrification activity. Enzyme subunits gene sequences annotation discovered that subunits of ammonia monooxygenase (amoA, amoB, amoC) and hydroxylamine oxygenase had higher expression levels compared with subunits of the other enzymes genes. Taxonomic profiles of selected enzymes (ammonia monooxygenase and hydroxylamine oxygenase) showed that ammonia-oxidizing bacteria present mainly belonged to Nitrosomonas and Nitrosospira species and no ammonia-oxidizing Archaea sequences were detected in both DNA and cDNA datasets.


Role of inhibitory and stimulative effects of prostaglandins on vasopressin-stimulated osmotic water flow in the toad bladder.
Vasopressin-prostaglandin (PG) interaction, especially the role of the inhibitory effects of PGE2 on vasopressin action, was studied using toad urinary bladders. The PGH2, at 1 X 10(-7) M, inhibited vasopressin-stimulated water flow (Marumo, 1982); PGE2 inhibited the water flow at 10(-8) M, but PGD2, PGF2 alpha, and PGI2 did not do so even at 10(-7) M. Thus, PGE2 has a physiological effect in contrast to other PGs converted from PGH2. Indomethacin enhanced both the vasopressin- and cyclic AMP-stimulated water flow across the toad bladder. However, the half maximum activation dose for vasopressin was 2 X 10(-10) M, but for cyclic AMP, as much as 3 X 10(-8) M. The PGE2 inhibited both vasopressin- and cyclic AMP-stimulated water flow. However, PGE2 inhibited vasopressin action in a dose-dependent manner which was not noted as a PGE2 effect on cyclic AMP action. The W-7, which is a specific inhibitor of calmodulin, suppressed cyclic AMP-stimulated water flow in a dose-dependent manner. Thus, PGE2 may suppress vasopressin-stimulated water flow at a site of cyclic AMP generation under physiological conditions. Thromboxane B2 (TXB2) enhanced vasopressin-stimulated water flow but not cyclic AMP-stimulated one. Thus PGE2 and TXB2 may be concluded as negative or positive modulators of vasopressin action in the toad bladder on the step(s) as the site of cyclic AMP generation under physiological conditions.


Nocturnal enuresis in adolescents with anorexia nervosa: prevalence, potential causes, and pathophysiology.
The aim of this study was to determine the prevalence and potential causes of secondary nocturnal enuresis (NE) in adolescents with anorexia nervosa (AN). Adolescents with AN completed the Incontinence Symptom Index-Pediatric (ISI-P), a self-report urinary incontinence survey. Those who had NE associated with the onset of AN on the ISI-P completed a comprehensive clinical and laboratory evaluation, including urinary flow measurements. The prevalence of NE in adolescents with AN was 17.0% (10 of 59). The overall frequency of day- and night-time urinary incontinence symptoms in adolescents with AN was 62.7%; urgency incontinence, 57.6%; stress incontinence, 32.2%; and insensate incontinence, 17.0%. Nine of the 10 adolescents with NE had secondary NE and confirmed that bedwetting ceased after weight restoration. Decreased functional bladder capacity and detrusor instability may contribute to the NE observed in this population. The high prevalence of NE in adolescents with AN warrants further study to determine the potential causative pathways. Clinicians who work with adolescents with AN should assess for this problem.


Effect of melatonin on monochromatic light-induced changes in clock gene circadian expression in the chick liver.
Light is the most prominent zeitgeber of the circadian system, which contains central and peripheral oscillators. Our previous studies found that light wavelength could influence the rhythms of melatonin synthesis and clock gene expression in the central oscillator of chicks. However, the effect of monochromatic light on the peripheral oscillator and the role of melatonin have yet to be clarified. In this study, 216 newly hatched chicks were divided into three groups (intact, sham operation and pinealectomy) and were raised under white (WL), red (RL), green (GL) or blue (BL) light for 14 days. Their plasma and livers were sampled at 6 time points with 4-h intervals. Plasma melatonin concentration and liver clock gene expression (cClock, cBmal1, cBmal2, cCry1, cCry2, cPer2, cPer3) were measured for circadian rhythm analysis. In intact and sham operation chicks under WL, all liver clock genes showed circadian expression along with oscillations in plasma melatonin. However, positive clock genes peaked at subjective night along with melatonin, while negative clock genes peaked at subjective day or the shifting time of day-night. Chick exposure to monochromatic light led to an unaltered circadian rhythmicity in plasma melatonin and liver clock genes; however, their rhythmic parameters were notably influenced. Compared to WL, GL enhanced the mesor and amplitude of melatonin and all kinds of clock genes, whereas RL had the opposite effect. Pinealectomy significantly decreased expression of liver clock genes, which was consistent with the reduction in plasma melatonin concentration, especially for the GL group, and resulted in the expression of liver clock genes showing low-mesor and low-amplitude oscillations as well as no statistically significant differences among the monochromatic light groups. Thus, we speculated that melatonin plays a key role in the effects of light wavelength on clock gene rhythm in the chick liver.


Effect of zinc, copper and glucocorticoids on metallothionein levels of cultured neurons and astrocytes from rat brain.
The knowledge of brain metallothionein (MT) regulation and especially of MT presence in specific cell types is scarce. Therefore, the effect of several well-known MT inducers, measured by radioimmunoassays using antibodies that cross-react with MT-I and MT-II or specific for MT-I and which do not cross-react with human growth inhibitory factor (GIF or MT-III), has been studied in primary cultures of neurons or astrocytes obtained from rat cerebrum. MT-I levels in glial cells were about ten times higher than those in neuronal cells (538 +/- 194 vs. 49 +/- 16 pg MT-I/micrograms protein, mean +/- S.D. from three separate cell preparations). Increasing the concentration of Zn in the bovine serum albumin (BSA)-containing culture medium up to 50 microM significantly increased MT-I levels by up to 3.5-fold in neurons and 2.5-fold in astrocytes. In contrast, Cu up to 50 microM increased MT-I levels in a saturable manner in both neurons (up to 5-fold) and astrocytes (up to 1.5-fold), the maximum effect occurring at 5 microM Cu. In general, the combination of Zn and Cu further increased MT-I levels. The effect of the metals on MT-I appeared to reflect metal uptake, since MT-I induction was less marked when the BSA concentration in the medium was increased from 2 to 10 mg/ml. Dexamethasone increased MT-I levels in both neurons and astrocytes in vitro in a concentration-dependent manner. Endotoxin, IL-1 and IL-6 did not have a significant effect on glial MT levels at the concentrations studied. The administration of dexamethasone to rats increased MT-I levels in non-frontal cortex, cerebellum, pons+medulla, midbrain and hippocampus, but not in hypothalamus, frontal cortex and striatum. Endotoxin increased liver but not brain MT-I levels. Immunocytochemical studies in adult rat brain preparations with a polyclonal antibody that cross-reacts with MT-I and MT-II indicated that immunostaining was always nuclear in glial cells, whereas in neurons it was nuclear in the cerebral cortex, hippocampus and the granular layer of the cerebellum, and nuclear plus cytoplasmic in Purkinje cells in the cerebellum, hypothalamic nuclei and gigantocellular reticular nucleus in the brain stem. Meninges, choroidal plexus, ependymal and endothelial cells were also MT-immunoreactive.


Preparation and applicability of Sarcocystis cruzi antigens and their anti-S. cruzi rabbit sera for serodiagnosis of bovine sarcocystosis.
Sarcocystis cruzi antigens and their antisera were prepared for the seroimmunological diagnosis of bovine sarcocystosis. Fresh S. cruzi cysts directly removed from cardial muscle of slaughtered cattle were digested with trypsin to release bradyzoites. By twelve cycles of freezing at-22 degrees C and thawing at 37 degrees C, bradyzoites were let to leach out soluble material. The soluble antigens were inoculated four times to rabbits at a dose of 343 micrograms protein and anti-S. cruzi sera were prepared with blood of the rabbits. Gel immunodiffusion test showed no cross-reaction between the present antigens and any of anti-Toxoplasma gondii, -Hammondia hammondi and -Besnoitia wallacei rabbit sera. Avidin-biotin complex immunoperoxidase (ABC) technique with the present anti-S. curzi rabbit sera showed clear positive reaction against S. cruzi cysts in the muscular sections of infected cattle.


Polarization-insensitive liquid crystal microlens array with dual focal modes.
We demonstrate a liquid crystal (LC) microlens array (MLA) fabricated by LCs possessing negative dielectric anisotropy, in conjunction with a cell with a three-electrode structure. The presented LC MLA is polarization-insensitive and can be operated in both concave and convex modes. The shortest focal length of the LC MLA is -2.54 and 2.22 mm in concave and convex mode, respectively. Disclination lines that are usually observed in conventional hole-patterned LC lens can also be avoided because of the vertical alignment treatment of LCs.


Epinastine inhibits eosinophil chemotaxis and adhesion molecules in atopic dermatitis.
To investigate the effects of epinastine on eosinophil chemotaxis and changes in eosinophil adhesion molecules induced by epinastine and three other antiallergic agents, using eosinophils of atopic dermatitis (AD) patients. Epinastine reduced eosinophil chemotaxis toward eotaxin when the eosinophils had been prestimulated with interleukin (IL)-5, but given alone it did not alter eosinophil chemotaxis toward IL-5. CD11b expression was inhibited when peripheral blood was prestimulated with IL-5, but eosinophil adhesion molecule expression was not altered. Epinastine suppresses allergic inflammation not only through its strong antihistamine and antimediator effects, but also by inhibiting eosinophilic chemotaxis and the expression of adhesion molecules involved in chemotaxis, especially CD11b.


Lumbar herniated disc treatment with percutaneous hydrodiscectomy.
Lumbar herniated disc are common manifestations of degenerative spine diseases, the main cause of radiated lower back pain. This guideline followed standard of a systematic review with recovery of evidence based on the movement of evidence-based medicine. We used the structured method for formulating the question synthesized by the acronym p.I.C.O., In which the p corresponds to the lumbar herniated disc, i to the treatment intervention with percutaneous hydrodiscectomy, c comparing with other treatment modalities, o the outcome of clinical evolution and complications. From the structured question, we identify the descriptors which constituted the evidence search base in the medline-pubmed databases (636 papers) and therefore, after the eligibility criteria (inclusion and exclusion), eight papers were selected to answer to clinical question. The details of the methodology and the results of this guideline are exposed in annex i.


Exploration of the molecular mechanism of prostate cancer based on mRNA and miRNA expression profiles.
Prostate cancer, the second most common cancer in men, has been rarely explored by integrating mRNA and miRNA expression profiles. In this study, we combined two mRNA expression datasets and six documented miRNAs to uncover the comprehensive molecular mechanism of prostate cancer. Results showed that a total of 30 genes were significantly differentially expressed in 49 tumor samples by comparing with 22 normal samples. Importantly, all samples from the two datasets can be clearly classified into two groups, tumor group and normal group, based on the selected differentially expressed genes (DEGs). The miRNA-mRNA regulation network indicated that 4 out of 30 DEGs can be regulated by three miRNAs. In addition, prognostic performance validation using in silico databases showed that the DEGs can significantly differentiate between low-risk and high-risk prostate cancer. In summary, multiple biological processes are probably involved in the development and progression of prostate cancer. First, dysregulation of SV2 can regulate transporter and transmembrane transporter activity and then provide the necessary nutrients for tumor cell proliferation. Second, HOXD10 can induce cell proliferation via TCF-4. Finally, dysregulation of CACNA1D can further suppress tumor apoptosis in prostate cancer. The identification of critical genes and valuable biological processes can be useful for the understanding of the molecular mechanism of prostate cancer.


Understanding the Density and Distribution of Restaurants in Los Angeles County to Inform Local Public Health Practice.
To describe the potential reach of restaurant-based strategies that seek to improve the healthfulness of menu options, it is important to understand the local restaurant environment, including the extent to which restaurants subject to policy mandates are located in communities disproportionately affected by diet-related diseases. This cross-sectional study examined the restaurant environment in Los Angeles County, a large jurisdiction with diverse geographic and socioeconomic characteristics, specifically 1) the number and characteristics of restaurants; 2) the association between neighborhood sociodemographics and restaurant density; and 3) the association between neighborhood sociodemographics and restaurant characteristics, including chain status (large chain, small chain, independent restaurant). Data sources were 1) industry data on restaurant location and characteristics (N = 24,292 restaurants) and 2) US Census data on neighborhood sociodemographics (N = 247 neighborhoods). We conducted descriptive and bivariate analyses at the restaurant and neighborhood level. Countywide, only 26.5% of all restaurants were part of a large chain (a chain with ≥20 locations). We found positive associations between restaurant density and neighborhood proportions of non-Hispanic white residents and residents with more than a high school education. We found limited support to suggest a greater density of large chains in neighborhoods with lower socioeconomic status. Results highlight the potentially limited reach of strategies targeting chain restaurants and point to the importance of including small chain restaurants and independent restaurants in public health efforts to improve the healthfulness of restaurants. Understanding where restaurants are in relation to priority populations is a critical step to planning strategies that address diet-related disparities.


The peristomial plates of ophiuroids (Echinodermata: Ophiuroidea) highlight an incongruence between morphology and proposed phylogenies.
The peristomial plates are skeletal components of the interbrachial frame (or mouth frame), which is located below the true mouth of ophiuroids. Whilst the peristomial plates were extensively described and used as diagnostic characters by some early workers, for the past 100 years they have been largely neglected as a taxonomic resource. In this investigation the peristomial plates of 48 species representing 21 families were examined directly, and information on a further 61 species, including representatives of another eight families, was obtained from the published literature. Observations were made with regard to fragmentation state, relative size and orientation of the peristomial plates. Although fragmentation state showed little consistency at any taxonomic level, relative size and orientation segregated a group of families comprising species with relatively small, inclined peristomial plates, viz. Ophiotrichidae, Ophiopholidae, Ophiactidae, Amphiuridae and Ophiocomidae, together with a single hemieuryalid species-Ophioplocus januarii. The distribution of peristomial plate traits was strongly correlated with that of several other character states pertaining to the interbrachial frame. This supported the proposition that two major types of interbrachial frame are present in ophiuroids (designated 'A' and 'B'). Current phylogenies inferred from both morphological and molecular data imply that type B is derived and has evolved independently at least twice in the orders Amphilepidida and Ophiacanthida. This represents a remarkable example of evolutionary convergence. An analysis of the distribution of all interbrachial frame character states suggested that within the Amphilepidida paedomorphosis was probably responsible for the complete reversion of the interbrachial frame to the ancestral type A condition in two families (Ophiothamnidae and Amphilepididae) of suborder Gnathophiurina and possibly responsible for varying degrees of trait reversal in the four families of suborder Ophionereidina. Such paedomorphic events may have been associated with a secondary return to the deep-sea from shallow-sea environments.


Autoradiographic quantification of 18F-FDG uptake in experimental soft-tissue abscesses in rats.
To use semiquantitative autoradiography to investigate fluorodeoxyglucose (FDG) uptake, distribution, and cellular localization in acute, early chronic, and late chronic soft-tissue infections. Unilateral calf-muscle abscesses were induced in 12 Sprague-Dawley rats by means of intramuscular inoculation of 0.1 mL of bacterial suspension (Staphylococcus aureus, 1.2 x 10(9) CFU/mL). Following injection of 130-180 MBq of fluorine 18 FDG, autoradiography of the abscess and contralateral muscle was performed (10-microm section thickness) on days 2, 5, and 9 after infection. Detailed spatial correlation of autoradiographs and histopathologic samples was performed by means of image fusion. Regions of interest were placed in the abscess wall, and measured gray values were converted to kilobecquerels per cubic centimeter according to kilobecquerels of injected activity per gram of body weight, which yielded standardized uptake values (SUVs). Acute abscess formation was characterized by central necrosis predominantly surrounded by neutrophils and a second necrotic tissue layer that bordered neutrophil infiltrates peripherally. Areas with increased FDG uptake corresponded to cellular inflammatory infiltrates, mainly granulocytes. The corresponding SUV was calculated to be 4.08 +/- 0.65 (mean +/- SD). Early chronic phase showed mixed cellular infiltrate of granulocytes and macrophages that surrounded central necrosis with interspersed fibroblasts and only residual muscle necrosis layer within the abscess wall. FDG uptake was located where granulocytes and macrophages were present, as in acute infection (SUV = 5.32 +/- 2.30). Late chronic infection was characterized by a prominent layer of macrophages around residual central necrosis and fibroblast-enriched granulation tissue delineating the infection from muscle tissue. FDG uptake clearly coincided with the macrophages, and no substantial increase of FDG uptake was detected within fibroblast-enriched granulation tissue. The SUV was calculated as 7.97 +/- 0.21. Results of Kruskal-Wallis ANOVA demonstrated that the change in SUV with time was statistically significant (chi(2) = 7.42, P <.05). The highest FDG uptake coincides with areas of inflammatory cell infiltrates, predominantly in neutrophils in the acute phase and in macrophages in the chronic phase of soft-tissue infection.


Camera array based light field microscopy.
This paper proposes a novel approach for high-resolution light field microscopy imaging by using a camera array. In this approach, we apply a two-stage relay system for expanding the aperture plane of the microscope into the size of an imaging lens array, and utilize a sensor array for acquiring different sub-apertures images formed by corresponding imaging lenses. By combining the rectified and synchronized images from 5 × 5 viewpoints with our prototype system, we successfully recovered color light field videos for various fast-moving microscopic specimens with a spatial resolution of 0.79 megapixels at 30 frames per second, corresponding to an unprecedented data throughput of 562.5 MB/s for light field microscopy. We also demonstrated the use of the reported platform for different applications, including post-capture refocusing, phase reconstruction, 3D imaging, and optical metrology.


Anti-atrial Fibrillation Effects of Pulmonary Vein Isolation With or Without Ablation Gaps: A Computational Modeling Study.
Although pulmonary vein isolation (PVI) gaps contribute to recurrence after atrial fibrillation (AF) catheter ablation, the mechanism is unclear. We used realistic computational human AF modeling to explore the AF wave-dynamic changes of PVI with gaps (PVI-gaps). We included 40 patients (80% male, 61.0 ± 9.8 years old, 92.5% persistent AF) who underwent AF catheter ablation to develop our realistic computational AF model. We compared the effects of a complete PVI (CPVI) and PVI-gap (2-mm × 4) on the AF wave-dynamics by evaluating the dominant frequency (DF), spatial change of DF, maximal slope of the action potential duration restitution curve (Smax), and AF defragmentation rate (termination or change to atrial tachycardia), and tested the effects of additional virtual interventions and flecainide on ongoing AF with PVI-gaps. Compared with the baseline AF, CPVIs significantly reduced extra-PV DFs (p < 0.001), but PVI-gaps did not. COV-DFs were greater after CPVIs than PVI-gaps (p < 0.001). Neither CPVIs nor PVI-gaps changed the mean Smax. CPVIs resulted in higher AF defragmentation rates (80%) than PVI-gaps (12.5%, p < 0.001). In ongoing AF after PVI-gaps, the AF defragmentation rates after a wave-breaking gap ablation, extra-PV DF ablation, or flecainide were 60.0, 34.3, and 25.7%, respectively (p = 0.010). CPVIs effectively reduced the DF, increased its spatial heterogeneity in extra-PV areas, and offered better anti-AF effects than extra-PV DF ablation or additional flecainide in PVI-gap conditions.


Effect of dissolved oxygen concentration on nitrite accumulation in nitrifying sequencing batch reactor.
A mathematical model based on Activated Sludge Model No. 3 (International Water Association, London) and laboratory-scale experiments were used to investigate ammonia conversion by nitrification in a sequencing batch reactor (SBR). The purpose of the study was to assess the effect of dissolved oxygen concentration on nitrite accumulation in the SBR. As the dissolved oxygen concentration in the SBR depends on the balance between oxygen consumption and oxygen transfer rates, ammonium conversion was measured for different air flowrate values to obtain different dissolved oxygen concentration profiles during the cycle. The ammonia concentration in the feeding medium was 500 mg ammonium as nitrogen (N-NH4(+))/L, and the maximum nitrite concentration achieved during a cycle was approximately 50 mg nitrite as nitrogen (N-NO2)/L. The air flow supplied to the reactor was identified as a suitable parameter to control nitrite accumulation in the SBR. This identification was carried out based on experimental results and simulation with a calibrated model. At a low value of the volumetric mass-transfer coefficient (kLa), the maximum nitrite concentration achieved during a cycle depends strongly on k(L)a, whereas, at a high value of k(L)a, the maximum nitrite concentration was practically independent of kL(a).


Kidney transplantation trends from UCLA registry Data, 1975-1982.
A total of 23,607 cases transplanted in 1975-1982 were analyzed for proportion and survival trends within eleven classification variables. Increases of up to 2% of total cases per year in proportions of registered transplants over the eight years are found in the following subcategories (with corresponding decreases in complementary subcategories): first grafts, cadaver donors, recipients with diabetes mellitus, and kidneys shipped more than 50 miles. Larger proportional increases of 3-7% per year are found for HLA-DR matching, cold ischemia times greater than 24 hr, cold storage, and pretransplant transfusions. Recipient population cross-sections are unchanged for age, race, HLA-A,B matching, and cytotoxic antibodies at transplant. Only the pretransplant transfusion classification has no increased graft survival in any subcategory; all other variables have one or more categories with increasing graft survival. It appears likely that the marked shift in transfusion policy nationwide has been the primary factor in increasing graft survival rates overall.


Parent Language Input Prior to School Forecasts Change in Children's Language-Related Cortical Structures During Mid-Adolescence.
Children differ widely in their early language development, and this variability has important implications for later life outcomes. Parent language input is a strong experiential factor predicting the variability in children's early language skills. However, little is known about the brain or cognitive mechanisms that underlie the relationship. In addressing this gap, we used longitudinal data spanning 15 years to examine the role of early parental language input that children receive during preschool years in the development of brain structures that support language processing during school years. Using naturalistic parent-child interactions, we measured parental language input (amount and complexity) to children between the ages of 18 and 42 months (n = 23). We then assessed longitudinal changes in children's cortical thickness measured at five time points between 9 and 16 years of age. We focused on specific regions of interest (ROIs) that have been shown to play a role in language processing. Our results support the view that, even after accounting for important covariates such as parental intelligence quotient (IQ) and education, the amount and complexity of language input to a young child prior to school forecasts the rate of change in cortical thickness during the 7-year period from 5½ to 12½ years later. Examining the proximal correlates of change in brain and cognitive differences has the potential to inform targets for effective prevention and intervention strategies.


Diverse morphology of biliary atresia in an animal model.
Extrahepatic biliary atresia can be simulated in Balb/c-mice which have been infected with rotavirus. Irreversible occlusion of the common bile duct is the result of an inflammatory process of the whole biliary tract. The observations in this animal model are similar to extrahepatic biliary atresia in newborn children. The present study describes the wide range of morphological findings in mice and compares the results with several classifications of extrahepatic biliary atresia in children. Newborn Balb/c-mice were infected intraperitoneally with rhesus rotavirus; the pathological morphology of the extrahepatic bile ducts in 73 mice is described and illustrated by scanning electron microscopy. The extension and localisation of atresia varied from short to interrupted or long-segment atresia, with or without prestenotic dilatation. The gallbladder was small and atretic, or appeared hydropic due to atresia of the common bile duct. The wide range of pathomorphological findings is the final stage of an inflammatory process. The morphological changes of the extrahepatic bile ducts do not fit any pattern, and no classification is evident. Most types of extrahepatic biliary atresia in children can be mimicked in this animal model. Comparing these observations with different classifications of extrahepatic biliary atresia in children, it must be asked if any classification of the disease is justified, having regard only to pathomorphological findings in the extrahepatic biliary tract.


The Depression Conundrum and the Advantages of Uncertainty.
According to the WHO (2012), the prevalence of unipolar depressive disorders is rising, even in those places where mental health treatments are widely available. The WHO predicts that these disorders will be the leading contributor to the global burden of disease by 2030. This sobering projection fits poorly with how psychological treatments for depression are presented in the mainstream scientific literature: as highly effective therapies, based upon a sound understanding of the causes of distress. There is a clear discrepancy between the rising prevalence figures on the one hand, and the confident claims of this effectiveness research on the other. This discrepancy prompts a set of complex interlinked questions, which we have called 'The Depression Conundrum.' In search of a partial answer, the aim of our study was to critically analyze five meta-analytic studies investigating the effectiveness of psychological EBTs for depression, all of which had been published in high impact factor journals. Our examination established a number of methodological and statistical shortcomings in every study. Furthermore, we argue that the meta-analytic technique is founded upon problematic assumptions. The implications of our analysis are clear: decades of quantitative research might not allow us to conclude that psychological EBTs for depression are effective. The uncertainty and questions raised by our findings might act as a catalyst to broaden the way in which depression and associated therapies are researched. In addition, it might contribute toward a more vigorous and interdisciplinary debate about how to tackle this soon-to-be global public health priority number one.


[[Ba2(C2O4)(H2O)6](NCS)2]n: a layered mixed-anion barium oxalate.
We present the first example of a compound containing Ba2+, C2O42-, water and some additional halide or pseudo-halide anions, viz. hexa-mu2-aqua-mu6-oxalato-dibarium(II) diisothiocyanate, [[Ba2(C2O4)(H2O)6](NCS)2]n. The structure consists of positively charged planar covalent layers of Ba2+ cations, oxalate anions and water molecules. The first coordination sphere of the Ba2+ cation contains six water molecules and four O atoms from two planar oxalate anions. The oxalate anion lies on an inversion centre and is coordinated to six Ba2+ cations, each donor O atom being bonded to two cations. Pairs of water molecules are coordinated by two Ba2+ cations. The layers are interspersed with non-coordinated NCS- anions.


Predictive value of serum gamma-glutamyltransferase levels in patients with hepatocellular carcinoma.
Here we assessed the predictive value of gamma-glutamyltransferase (γ-GT) for the prognosis of patients with HCC and compared the γ-GT with other prognostic factors. We retrospectively analyzed outcomes for 858 patients first diagnosed with HCC. Cox univariate and multivariate analyses receiver operating characteristic (ROC) curve were used for the study of significance of prognostic factor. A Kaplan-Meier survival analysis was performed to assess the value of γ-GT as an HCC prognostic factor in different classifications of Barcelona Clinic Liver Cancer (BCLC) or Tumor Node Metastasis (TNM) and different levels of serum alpha fetoprotein (AFP). We showed patient survival rates were significantly associated with γ-GT as well as serum biological markers including absolute neutrophil count (ANC), absolute lymphocyte count (ALC), AFP. A γ-GT ≥ 75 U/L strongly indicated poor prognosis for HCC patients. The survival time of patients with γ-GT ≥ 75 U/L was significantly shorter in advanced BCLC and TNM stages and at any serum AFP level. All these results suggested that baseline γ-GT could effectively aid in determining the prognosis of patients with HCC, and the prognostic value of γ-GT ≥ 75 U/L was superior to that of Child-Pugh class, MELD stage, and serum AFP.


Electrocerebral inactivity associated with obstructive sleep apnea.
We report a child who demonstrates near electrocerebral silence secondary to obstructive apnea during polysomnography. The mechanism of this suppression of cortical activity appears to be related to hypoxemia in the absence of a malignant arrhythmia.


Uncertain LDA: Including Observation Uncertainties in Discriminative Transforms.
Linear discriminant analysis (LDA) is a powerful technique in pattern recognition to reduce the dimensionality of data vectors. It maximizes discriminability by retaining only those directions that minimize the ratio of within-class and between-class variance. In this paper, using the same principles as for conventional LDA, we propose to employ uncertainties of the noisy or distorted input data in order to estimate maximally discriminant directions. We demonstrate the efficiency of the proposed uncertain LDA on two applications using state-of-the-art techniques. First, we experiment with an automatic speech recognition task, in which the uncertainty of observations is imposed by real-world additive noise. Next, we examine a full-scale speaker recognition system, considering the utterance duration as the source of uncertainty in authenticating a speaker. The experimental results show that when employing an appropriate uncertainty estimation algorithm, uncertain LDA outperforms its conventional LDA counterpart.


[SATISFACTION OF YOUNG DOCTORS IN CROATIA: ARE WE HEADING IN THE RIGHT DIRECTION]?
The satisfaction of junior medical doctors is primarily associated with the training they receive during the residency period and working conditions. Given a considerable brain-drain of medical doctors from the Republic of Croatia, the present research focused on the evaluation of the satisfaction of junior medical doctors. The Junior Doctors Committee of the Croatian Medical Chamber prepared a questionnaire on the satisfaction of junior doctors. The relevant questionnaire was available for filling in online in the period from February 1 to March 20, 2016. The questionnaire was completed by 1,531 persons aged between 29 and 35 (mean age 32) of which 67% were females. A 58% of respondents would leave the Republic of Croatia if offered an opportunity. The main reasons for leaving the country include better working conditions (74%), well-regulated healthcare system (64%) and higher wages (64%). In case of staying in the country, their future professional status and development would remain unaltered (44%). The questionnaire results show a high level of dissatisfaction with the healthcare system, some segments of the specialist training and the mentor role. The analysis of factors affecting the satisfaction of junior doctors will facilitate the preparation and adoption of measures aiming to mitigate the above-mentioned trend.


Rapid and accurate synthesis of TALE genes from synthetic oligonucleotides.
Custom synthesis of transcription activator-like effector (TALE) genes has relied upon plasmid libraries of pre-fabricated TALE-repeat monomers or oligomers. Here we describe a novel synthesis method that directly incorporates annealed synthetic oligonucleotides into the TALE-repeat units. Our approach utilizes iterative sets of oligonucleotides and a translational frame check strategy to ensure the high efficiency and accuracy of TALE-gene synthesis. TALE arrays of more than 20 repeats can be constructed, and the majority of the synthesized constructs have perfect sequences. In addition, this novel oligonucleotide-based method can readily accommodate design changes to the TALE repeats. We demonstrated an increased gene targeting efficiency against a genomic site containing a potentially methylated cytosine by incorporating non-conventional repeat variable di-residue (RVD) sequences.


Double-Blind, Placebo-Controlled, Crossover Study of Armodafinil Treatment of Daytime Sleepiness Associated With Treated Nocturia.
Nocturia, voids which disturb sleep, is the most common cause of awakenings and is associated with daytime sleepiness. Because the standard treatments for the most common causes of nocturia are relatively ineffective, many treated patients with nocturia are left with residual sleepiness. We carried out this pilot study to evaluate the potential of armodafinil to be an effective means of addressing the sleepiness that persists in many nocturia patients, despite their receiving standard therapy. This was a double-blind, placebo-controlled, crossover study carried out in 28 patients with nocturia who were receiving standard clinical therapy for their nocturia and who had an Epworth Sleepiness Scale (ESS) score of at least 10. Subjects received 4 weeks of both armodafinil (150-250 mg) and placebo with order randomized. Armodafinil led to statistically significant improvement in sleepiness compared to placebo as indicated by the ESS (the primary outcome; p < .002) as well as the Clinical Global Impression of Improvement in Sleepiness scale (key secondary outcome; p = .01). Armodafinil did not increase nocturic events or significantly increase adverse effects versus placebo. This pilot study, the first double-blind, placebo-controlled trial assessing whether a wake-promoting therapy can improve residual daytime sleepiness in patients with treated nocturia, indicates the promise of armodafinil for addressing this residual sleepiness and provides impetus to carry out a large-scale study to definitively evaluate whether armodafinil is an effective therapy for the many patients with nocturia who experience daytime sleepiness that persists, despite their receiving standard therapy for this condition.


Case Report: Fibromatosis colli in a neonate.
Fibromatosis colli or pseudotumor of infancy of the sternocleidomastoid muscle is a rare cause of a benign neck mass in neonates and infants. If diagnosed correctly, it can be managed conservatively, and unnecessary investigations can be avoided.


Conduction apraxia.
A left hemisphere damaged patient with ideomotor apraxia is described, whose performance on pantomime to verbal command was superior to pantomime imitation. His reception of these same gestures (gesture naming) was spared. This syndrome has been named conduction apraxia. To account for this selective impaired performance on gesture imitation, a separation of the representations for gesture production and reception is proposed and a non-lexical gesture processing route for gesture imitation is suggested.


Measurement of free erythrocyte protoporphyrin in blood collected on filter paper as a screening test to detect lead poisoning in children.
A procedure for the measurement of free erythrocyte protoporphyrin (FEP) in a drop of blood collected on filter paper is described. The method is useful as a screening test for lead poisoning in children. Based on the FEP finding and blood lead tests, asymptomatic children are classified into four major categories. A course of action is suggested for each category.


[Perioperative management of airway foreign bodies in 35 pediatric patients].
We retrospectively reviewed 33 patients (35 cases) who underwent foreign body removal at our institution from 1995 through 2003. Male-female ratio was 21 : 12 and the most frequent age was one year. The most common foreign bodies (FBs) were nuts (n = 14) and plastics (n = 7). A repeater (3 cases) had mental retardation. Patients were referred to our institution with an average interval of 90 hours, and after 1.3 hospitals. All the patients were managed with general anesthesia. Direct laryngoscopy was performed to extract FBs in 11 cases suspected of having pharyngeal or laryngeal FBs. In 24 cases suspected of having tracheobronchial FBs, the trachea was intubated and a flexible fiberoptic bronchoscopy was performed to locate the FBs. FBs were found in the trachea in 2 cases and in the bronchus in 18 patients and were successfully extracted by rigid bronchoscopy in 10 cases. All the patients were admitted for fear of laryngotracheal edema. There were no significant postoperative complications.


Tracking sleep in the field: sleep quality and sleep behaviours of elite track and field athletes during preparation and competition.
Elite athletes are susceptible to inadequate sleep, which may peak during competition and be exacerbated by poor sleep behaviours. This study sought to characterise and compare the sleep quality and sleep behaviours of elite track and field athletes during preparation and major competitions. Forty elite international track and field athletes (50% female, aged 25.1 ± 3.9 years) completed the Athlete Sleep Screening Questionnaire and the Athlete Sleep Behaviour Questionnaire on three separate occasions: during habitual training, during a pre-meet training camp and during a major international competition. Overall, 62.5% of athletes reported at least mild sleep difficulty during competition. Athletes reported higher sleep difficulty and poorer sleep behaviour during major competitions and the pre-meet training camp compared to habitual training (P = .001-.025). No significant differences were observed between the training camp and major competition. Global sleep behaviour scores were underpinned by unique characteristics at each timepoint. Sleep behaviour (R2 = .330, P = .017), injury status (R2 = .253, P = .003) and major championship experience (R2 = .113, P = .034) were associated with sleep difficulty during competition. Sleep quality and behaviours vary according to stage of the track and field season, providing a foundation for targeted intervention.


Adoption and Implementation of Evidence-Based Colorectal Cancer Screening Interventions Among Cancer Control Program Grantees, 2009-2015.
Colorectal cancer (CRC) is the second-leading cause of cancer death in the United States. Although effective CRC screening tests exist, CRC screening is underused. Use of evidence-based interventions (EBIs) to increase CRC screening could save many lives. The Colorectal Cancer Control Program (CRCCP) of the Centers for Disease Control and Prevention (CDC) provides a unique opportunity to study EBI adoption, implementation, and maintenance. We assessed 1) the number of grantees implementing 5 EBIs during 2011 through 2015, 2) grantees' perceived ease of implementing each EBI, and 3) grantees' reasons for stopping EBI implementation. CDC funded 25 states and 4 tribal entities to participate in the CRCCP. Grantees used CRCCP funds to 1) provide CRC screening to individuals who were uninsured and low-income, and 2) promote CRC screening at the population level. One component of the CRC screening promotion effort was implementing 1 or more of 5 EBIs to increase CRC screening rates. We surveyed CRCCP grantees about EBI implementation with an online survey in 2011, 2012, 2013, and 2015. We conducted descriptive analyses of closed-ended items and coded open-text responses for themes related to barriers and facilitators to EBI implementation. Most grantees implemented small media (≥25) or client reminders (≥21) or both all program years. Although few grantees reported implementation of EBIs such as reducing structural barriers (n = 14) and provider reminders (n = 9) in 2011, implementation of these EBIs increased over time. Implementation of provider assessment and feedback increased over time, but was reported by the fewest grantees (n = 17) in 2015. Reasons for discontinuing EBIs included funding ending, competing priorities, or limited staff capacity. CRCCP grantees implemented EBIs across all years studied, yet implementation varied by EBI and did not get easier with time. Our findings can inform long-term planning for EBIs with state and tribal public health institutions and their partners.


Cluster-seeded synthesis of doped CdSe:Cu4 quantum dots.
We report here a method for synthesizing CdSe quantum dots (QDs) containing copper such that each QD is doped with four copper ions. The synthesis is a derivative of the cluster-seed method, whereby organometallic clusters act as nucleation centers for quantum dots. The method is tolerant of the chemical identity of the seed; as such, we have doped four copper ions into CdSe QDs using [Na(H2O)3]2[Cu4(SPh)6] as a cluster seed. The controlled doping allows us to monitor the photophysical properties of guest ions with X-ray spectroscopy, specifically XANES and EXAFS at the copper K-edge. These data reveal that copper can capture both electrons and holes from photoexcited CdSe QDs. When the dopant is oxidized, photoluminescence is quenched and the copper ions translocate within the CdSe matrix, which slows the return to an emissive state.


Relationships between milk urea and production, nutrition, and fertility traits in Israeli dairy herds.
The objectives of this study were to identify and evaluate production and environmental factors that influence milk urea (MU) in Israeli dairy herds, to analyze the relationships between MU concentration and nutritional variables, and to examine a possible association between MU and pregnancy rate (PR). Production and environmental data were obtained from the Israeli Dairy Herd Improvement (DHI) Center (n = 1,279,600). Programmed total mixed rations (feeds and quantities) on milk-test day were collected from 42 dairy herds. Data on 36,073 cows that were inseminated close to milk-test day and pregnancy diagnosis results were obtained from the DHI data bank. Highly significant positive relationships were found between MU concentration and milk yield and fat percentage; relationships between MU and milk total protein percentage and somatic cell count were negative. Milk urea levels were higher during the summer months and were higher for adult cows. These levels increased as lactation progressed. Milk urea was positively associated with dietary levels of crude protein, ruminal digestible protein, and neutral detergent fiber contents; it was negatively associated with ration energy and nonstructural carbohydrate contents. Significant influences of specific feeds on MU were detected. A significant negative association was found between MU level and PR. Least squares means for PR for cows in the lowest and highest MU quartiles were 38.4 and 36.1%, respectively. Increasing levels of MU were negatively related to reproductive performance of dairy cows, but the risk of nonpregnancy caused by high levels of MU was lower than reported in previous studies.


Parent-Mediated Interventions for Infants under 24 Months at Risk for Autism Spectrum Disorder: A Systematic Review of Randomized Controlled Trials.
Prodromal symptoms of Autism Spectrum Disorder (ASD) have been detected within the first year of life. This review evaluated evidence from randomized controlled trials (RCTs) of parent-mediated interventions for infants under 24 months who are at risk for ASD. Electronic databases, including grey literature, were searched up till November 2019. Seven RCTs were identified. There was substantial heterogeneity in recruitment, outcome measures and effect size calculations. Interventions did not reduce the risk of later ASD diagnosis and post-intervention effects on infant outcomes were inconsistent, with five studies reporting significant improvements across both treatment and control groups. Moderate level of evidence of intervention effects on parental interaction skills and the small number of RCTs, and significant limitations restrict generalizability across studies.


Triton X-100 or octyl glucoside inactivates acyl-CoA:cholesterol acyltransferase 1 by dissociating it from a two-fold dimer to a two-fold monomer.
Cholesterol is an important lipid molecule and is needed for all mammalian cells. In various cell types, excess cholesterol is stored as cholesteryl esters; acyl-CoA:cholesterol acyltransferase 1 (ACAT1) plays an essential role in this storage process. ACAT1 is located at the endoplasmic reticulum and has nine transmembrane domains (TMDs). It is a member of the membrane-bound O-acyltransferase (MBOAT) family, in which members contain multiple TMDs and participate in a variety of biological functions. When solubilized in the zwitterionic detergent CHAPS, ACAT1 can be purified to homogeneity with full enzyme activity and behaves as a homotetrameric protein. ACAT1 contains two dimerization motifs. The first motif is located near the N-terminus and is not conserved in MBOATs. Deletion of the N-terminal dimerization domain converts ACAT1 to a dimer with full catalytic activity; therefore, ACAT1 is a two-fold dimer. The second dimerization domain, located near the C-terminus, is conserved in MBOATs; however, it was not known whether the C-terminal dimerization domain is required for enzyme activity. Here we show that treating ACAT1 with non-ionic detergent, Triton X-100 or octyl glucoside, causes the enzyme to become a two-fold monomer without any enzymatic activity. Detergent exchange of Triton X-100 with CHAPS restores ACAT1 to a two-fold dimer but fails to restore its enzymatic activity. These results implicate that ACAT1 requires hydrophobic subunit interactions near the C-terminus in order to remain active as a two-fold dimer. Our results also caution the use of Triton X-100 or octyl glucoside to purify other MBOATs.


Bactericidal effects of mouth rinses on oral bacteria.
The bactericidal efficacy of two types of Listerine; Listerine and Cool Mint Listerine, and povidone iodine on oral microorganisms, methicillin-resistant Staphylococcus aureus (MRSA), Streptococcus pyogenes, Helicobacter pylori and Candida albicans were examined. Most of the oral bacteria were killed completely by a 10-sec exposure to Listerine or Cool Mint Listerine. H. pylori, MRSA and C. albicans were also reduced by a 30-sec exposure to the Listerine mouth rinse. Bacteria in dental plaque were decreased by exposure to Listerine, Cool Mint Listerine, and povidone iodine for 30 seconds. Mouthwashing with Listerine for 30 seconds resulted in a decrease to approximately 1/100 of the viable bacterial counts in saliva. These bactericidal effects against bacteria in saliva and dental plaque indicated that Listerine and Cool Mint Listerine antiseptic are useful in oral cavity as antiseptic mouth rinses.


Optimization of combined microwave and hot air drying technology for purple cabbage by Response Surface Methodology (RSM).
Purple cabbage is one of the world's most widely consumed vegetables with high nutritional values containing the antioxidants and anti-inflammatory activity of anthocyanins, vitamins, and minerals. But due to rapid postharvest quality decay, novel processing techniques including drying are required. In order to determine the conditions of combined microwave and hot air drying for purple cabbage, factors affecting the drying process including microwave density, hot air temperature, and the dry base water content at conversion point were investigated using the anthocyanin content, DPPH antioxidant capacity, chewiness, △E, rehydration ratio, and average drying rate as responses. The combined drying conditions were optimized considering three independent variables at three different levels by response surface methodology. The results showed that the processing parameters of purple cabbage with combined microwave and hot air drying technology were microwave density at 2.5 W/g, moisture content of conversion point at 4.0 g/g, and hot air temperature at 55°C. Under these conditions, the anthocyanin content, DPPH antioxidant capacity, chewiness, △E, rehydration ratio, average drying rate, and overall score of the dried purple cabbage were 175.87 mg/100 g, 87.59%, 4,521.468 g, 26.5, 4.3, 0.76 g/min, and 0.785, respectively. Therefore, combined microwave and hot air drying technology is an effective, suitable method for drying purple cabbage.


Enhanced optical activity and circular dichroism in twisted photonic crystal fiber.
We demonstrate experimentally and theoretically that the core-guided mode in helically twisted photonic crystal fiber exhibits resonantly enhanced optical activity and circular dichroism in the vicinity of anti-crossings with leaky orbital angular momentum (OAM) modes in the cladding. This arises because the anti-crossings for left and right circularly polarized core modes occur at slightly different wavelengths.


Claudin-7 promotes the epithelial-mesenchymal transition in human colorectal cancer.
In colorectal cancer (CoCa) EpCAM is frequently associated with claudin-7. There is evidence that tumor-promoting EpCAM activities are modulated by the association with claudin-7. To support this hypothesis, claudin-7 was knocked-down (kd) in HT29 and SW948 cells. HT29-cld7kd and SW948-cld7kd cells display decreased anchorage-independent growth and the capacity for holoclone-, respectively, sphere-formation is reduced. Tumor growth is delayed and cld7kd cells poorly metastasize. In line with this, migratory and invasive potential of cld7kd clones is strongly impaired, migration being inhibited by anti-CD49c, but not anti-EpCAM, although motility is reduced in EpCAM siRNA-treated cells. This is due to claudin-7 recruiting EpCAM in glycolipid-enriched membrane fractions towards claudin-7-associated TACE and presenilin2, which cleave EpCAM. The cleaved intracellular domain, EpIC, promotes epithelial-mesenchymal transition (EMT)-associated transcription factor expression, which together with fibronectin and vimentin are reduced in claudin-7kd cells. But, uptake of HT29wt and SW948wt exosomes by the claudin-7kd lines sufficed for transcription factor upregulation and for restoring motility. Thus, claudin-7 contributes to motility and invasion and is required for recruiting EpCAM towards TACE/presenilin2. EpIC generation further supports motility by promoting a shift towards EMT. Notably, EMT features of cld7-competent metastatic CoCa cells can be transferred via exosomes to poorly metastatic cells.


Ti-Catalyst Biomimetic Sensor for the Detection of Nitroaromatic Pollutants.
An electrode modified by a Ti complex in Nafion was used in the electrochemical detection of nitroaromatic compounds (NACs). The catalyst reduced nitroaromatic groups by a six-electron process with a high apparent rate constant, k1, which was estimated to be at least 19 000 mol-1 L s-1 by simulation of the first step of the cathodic reduction of 4-nitrophenylacetic acid, which was used as model molecule. The modified electrode was prepared and optimized in terms of Nafion and catalyst concentrations. In the chosen analytical medium, the Ti-catalyst-containing electrode showed good stability in the presence of nitro species. A standard deviation around 15% was calculated on the analysis performed with 28 different modified electrodes, which could be expected for a drop-coating process. We show that the modified electrode detects several nitroaromatic compounds such as nitrophenols, mononitroaniline, and dinitrotoluene with detection limits ranging from 1 × 10-4 to 9 × 10-4 g L-1 (0.2 × 10-6 to 5.1 × 10-6 mol L-1). As an interesting feature, the sensor exhibits good selectivity toward NAC detection because titanocene does not catalyze the reduction of nitroaliphatic compounds.


Laser emission from tapered fiber-based liquid-crystal microsphere for sensing.
This Letter introduces a novel laser emission probe for liquid-crystal microspheres based on a tapered fiber. A cholesteric liquid crystal (CLC) is injected into a hollow glass microsphere (HGM) attached at the front end of a tapered fiber in order to produce laser. Tapered fibers are preferable to rectangular fibers for liquid-crystal microsphere laser emission. The whispering gallery mode (WGM) laser is significantly suppressed by the tapered fiber-based liquid-crystal microsphere, which also displays an apparent single-mode photonic bandgap (PBG) laser peak. The stimulation response of tapered fiber-based liquid-crystal microspheres to organic vapors causes a modification of the laser peak wavelength with increasing gas concentration. In addition, laser emission generated by tapered fiber-based liquid-crystal microspheres is expected to be used in fields such as microenvironmental biosensing.


Impact of a harm-reduction policy on HIV and hepatitis C virus transmission among drug users: recent French data--the ANRS-Coquelicot Study.
In France a harm-reduction policy was implemented in the late 1980s with the aim of reducing the prevalence of HIV and hepatitis C virus (HCV) infection among drug users. The ANRS-Coquelicot survey was designed to measure the prevalence of HIV and HCV infection among drug users and to examine determinants of at-risk behaviors. In 2002, information was collected from 166 drug users recruited in all types of services specializing in drug use intervention and harm reduction in Marseille, France. Self-reported HIV and HCV serostatus was compared with the results of serological tests done on capillary blood collected on filter paper. The self-reported and biologically documented prevalence rates of HIV infection were identical (22%). In contrast, the self-reported prevalence of HCV infection was 52%, whereas the biologically documented prevalence was 73%. Overall, 30% of HCV-infected drug users were unaware of their status. Forty-four percent of drug users under 30 years of age were HCV seropositive, suggesting that they had been infected early during drug use. The harm-reduction policy seems to have had a marked impact on HIV transmission among drug users but a much more limited impact on HCV transmission. The limitations and implications of the study are discussed.


Plasticity of mesangial cells: a basis for understanding pathological alterations.
In the last two decades, the ability of mesangial cells to respond to various stimuli or injurious agents by altering their phenotype and function has become recognized. The plasticity of these mesangial cells has been linked to the morphological and functional alterations responsible for the pathologic findings. Many of the glomerular disorders target the mesangium as the primary and/or initial site of injury. Understanding how mesangial cells are altered in the various conditions provides a platform for conceptualizing pathologic mechanisms and defining key steps amenable to therapeutic intervention. The present paper reviews the normal and altered mesangium with an emphasis on mechanisms involved in alterations of mesangial homeostasis. Mesangial cells and matrix are very important in maintaining normal glomerular structure, and function and the plasticity of these cells is responsible for pathological manifestations, repair, and scarring. Our more sophisticated understanding of mesangial cell behavior and matrix biology provides very useful information to help design new therapeutic approaches to the treatment of renal diseases. The potential for bone marrow-derived cells to differentiate into mesangial cells and repopulate damaged mesangium, thus "healing" what is today considered to be irreversible damage represents an exciting new area of research.


Id2 complexes with the SNAG domain of Snai1 inhibiting Snai1-mediated repression of integrin β4.
The epithelial-mesenchymal transition (EMT) is a fundamental process that underlies development and cancer. Although the EMT involves alterations in the expression of specific integrins that mediate stable adhesion to the basement membrane, such as α6β4, the mechanisms involved are poorly understood. Here, we report that Snai1 inhibits β4 transcription by increasing repressive histone modification (trimethylation of histone H3 at K27 [H3K27Me3]). Surprisingly, Snai1 is expressed and localized in the nucleus in epithelial cells, but it does not repress β4. We resolved this paradox by discovering that Id2 complexes with the SNAG domain of Snai1 on the β4 promoter and constrains the repressive function of Snai1. Disruption of the complex by depleting Id2 resulted in Snai1-mediated β4 repression with a concomitant increase in H3K27Me3 modification on the β4 promoter. These findings establish a novel function for Id2 in regulating Snai1 that has significant implications for the regulation of epithelial gene expression.


Nifedipine therapy for patients with threatened and acute myocardial infarction: a randomized, double-blind, placebo-controlled comparison.
Preliminary clinical and laboratory observations suggest that nifedipine might prevent progression of threatened myocardial infarction by reversing coronary spasm or might limit necrosis during the course of acute myocardial infarction. We screened 3143 patients with ischemic pain of greater than 45 min duration and randomly assigned 105 eligible patients with threatened myocardial infarction and 66 with acute myocardial infarction to receive nifedipine (20 mg orally every 4 hr for 14 days) or placebo plus standard care. Treatment was started 4.6 +/- 0.1 hr after the onset of pain. Infarct size index was calculated by the MB-creatine kinase (CK) method and expressed as CK-geq/m2 +/- SE. The incidence of progression to infarction among patients with threatened myocardial infarction was not significantly altered by nifedipine (36 of 48 [75%] for placebo-treated and 43 of 57 [75%] for nifedipine-treated patients). Furthermore, infarct size index was similar among placebo- and nifedipine-treated patients (16.9 +/- 1.5 MB-CK-geq/m2, n = 65, and 17.0 +/- 1.5 MB-CK-geq/m2, n = 68, respectively) with threatened myocardial infarction who exhibited infarction and for those with acute myocardial infarction. Among the 171 eligible patients randomly assigned to drug or placebo, 6 month mortality did not differ significantly (8.5% for placebo vs 10.1% for nifedipine, NS), but mortality in the 2 weeks after randomization was significantly higher for nifedipine-treated patients (0% for placebo compared with 7.9% for nifedipine, p = .018).(ABSTRACT TRUNCATED AT 250 WORDS)


A sequence of biochemical events in the antigen-induced release of chemical mediators from sensitized human lung tissue.
Five sequential steps interspaced between the antigen activation of human lung fragments sensitized with IgE and the release of the chemical mediators, histamine and slow-reacting substance of anaphylaxis (SRS-A), have been delineated. The experimental design that permits this analysis is based upon the capacity to maintain the serine esterase essential to mediator release in its diisopropylphosphofluoridate (DFP)-resistant precursor state despite antigen challenge and upon the ability to arrest reversibly the reaction sequence by various manipulations. When sensitized lung fragments are challenged with antigen in the presence of DFP, a serine esterase is converted to its active DFP-inhibitable state; this conversion is prevented if antigen challenge in the presence of DFP occurs in calcium-free buffer indicating that immunologic activation of the esterase requires extracellular calcium. The fact that calcium depletion alone does not impair antigen-induced histamine release implies that prevention of esterase activation depends upon both the absence of extracellular calcium and the inactivation of any active esterase by DFP to prevent an autocatalytic feedback activation. Arresting the antigen-induced activation of the serine esterase by the combination of DFP in calcium-free buffer precludes the sequence from reaching the labile, 2-deoxyglucose (2-DG)-inhibitable, energy-requiring step, indicating that proesterase activation precedes this energy-requiring stage. The 2-DG-inhibitable step precedes a second calcium-requiring, EDTA-inhibitable stage, as EDTA prevents glucose reversal of 2-DG inhibition of antigen-challenged tissue, while the presence of 2-DG does not prevent calcium reversal of EDTA inhibition. The finding that isoproterenol prevents calcium reversal of EDTA inhibition of mediator release suggests that the inhibitory site of action of increased concentrations of cyclic AMP is coincident with or subsequent to the second calcium-requiring, EDTA-inhibitable step. Therefore, the sequence of biochemical events initiated by the interaction of antigen with tissue-fixed IgE antibodies appears to proceed from the calcium-requiring activation of a DFP-sensitive serine esterase; the further autocatalytic activation of the esterase; a 2-DG-inhibitable energy requirement; a second calcium-requiring, EDTA-inhibitable stage; and a cyclic AMP-inhibitable step to the release of histamine and SRS-A.


Localization of IgE synthesis in immediate-type allergy of the upper respiratory tract.
The localization of allergen-specific IgE synthesis in allergic diseases of the upper respiratory tract is so far unknown. It has been suggested that the IgE production takes place in the nasal mucosa itself. The present immunohistochemical studies with anti-IgE and monoclonal markers for B lymphocytes, plasma cells, antigens of the major histocompatibility complex, T helper and T suppressor cells indicate that there are no IgE-producing plasma cells in the nasal mucosa of patients with seasonal allergic rhinitis. The IgE-associated cells have been defined as two different types of mast cells. Furthermore we found IgE-associated lymphoid follicles in palatine as well as in nasopharyngeal tonsils and in cervical lymph nodes from allergic patients. IgE-specific activated B lymphocytes and plasma cells were identified in direct contact with migratory mast cells in these lymphoid tissues. We therefore suggest that IgE synthesis takes place in the lymphoid tissues of Waldeyer's ring and in downstream cervical lymph nodes and that migratory mast cells transport specific IgE back to the nasal mucosa to mediate the allergic reaction.


Case report: Long-term survival in a patient with metastatic colorectal cancer treated with trifluridine/tipiracil in the third-line setting.
Third-line treatment with trifluridine/tipiracil (FTD/TPI) is recommended for patients with metastatic colorectal cancer (mCRC) or gastric/gastroesophageal cancer (GC) who have progressed beyond first- and second-line therapy. We describe a patient with long-term survival following treatment with FTD/TPI. The patient, a 70-year-old woman diagnosed with right-sided mCRC (T3/N1) with metastases to the aortocaval and precaval lymph nodes, received first-line panitumumab and capecitabine for 6 months, followed by second-line bevacizumab and oxaliplatin. She had disease progression following 9 months of second-line therapy and began third-line treatment with FTD/TPI (50 mg bid). Three months after treatment initiation, lymph node involvement was reduced, and following 12 months of FTD/TPI treatment, her disease had stabilized, and she reported no treatment-related adverse events. She remained on the same dose of FTD/TPI for more than 27 months after initiating treatment, with maintenance of stable disease. This patient with mCRC demonstrated a survival benefit with FTD/TPI beyond those reported in published clinical trial data and real-world studies.


Research and its relationship to nurse education: focus and capacity.
Nursing, midwifery and health visiting together form the largest professional workforce within the NHS, and the clinical activities of nurses and the outcomes of these interventions are, therefore, of critical importance to patients. In consequence, adequate preparation for practice should include evidence-based nursing interventions, education to evaluate such evidence, understanding of research methodology and exposure to educators who are engaged in clinical research and can thus act as appropriate models for learners who may aspire to clinical research careers in the future. More broadly, the achievement of these goals requires a continuing strategy to develop capacity for the understanding, undertaking, dissemination and utilisation of research within the profession. This paper argues that nursing is currently in a weak position with regard to creating a future workforce which will have this capacity. Examination of two mental health journals illustrates the lack of focus on clinical research within the profession, whilst a key Government document setting out the future of nursing contains little reference to the role of R&D either in clinical practice or the preparation of the workforce. A recent publication setting out a potential strategy for research in nursing contains considerable hope for the future, but the profession will wish to remain watchful to ensure that the document is translated into action to facilitate a future for clinical nursing research appropriate to the needs of patients.


Adhesion of Toxoplasma gondii tachyzoite-infected vehicle leukocytes to capillary endothelial cells triggers timely parasite egression.
Intracellular pathogens have numerous strategies for effective dissemination within the host. Many intracellular pathogens first infect leukocytes, which they use as a vehicle to transport them to target organs. Once at the target organ, intracellular parasite Toxoplasma gondii can cross the capillary wall in extracellular form by infecting endothelial cells. However, after egression from leukocytes, extracellular parasites face the risk of host immune attack. In this study, observation of infected mouse organs, using a method that renders tissue transparent, revealed that adhesion of tachyzoite-infected leukocytes to endothelial cells triggers immediate egression of the parasite. This signal enables the parasite to time egression from its vehicle leukocyte to coincide with arrival at a target organ, minimizing the opportunity for immune attack during the transition from a vehicle leukocyte to capillary endothelial cells.


Development and evaluation of a real-time PCR assay for detection of Klebsiella pneumoniae carbapenemase genes.
We developed a novel real-time PCR assay to detect Klebsiella pneumoniae carbapenemases (KPCs) and used this assay to screen clinical isolates of K. pneumoniae and Klebsiella oxytoca for the presence of bla(KPC) genes. The TaqMan real-time PCR assay amplified a 399-bp product from the bla(KPC) gene. The amplicon was designed so that the genes for isoenzymes KPC-1, -2, and -3 could be easily distinguished by subsequent restriction digestion of the amplicon with the enzymes BstNI and RsaI. The assay was validated with reference strains obtained from the Centers for Disease Control and Prevention that contained each of the three described isoenzymes and 69 extended-spectrum beta-lactamase-producing clinical isolates (39 K. pneumoniae and 30 K. oxytoca isolates). Subsequently, the bla(KPC) PCR assay was used to confirm the presence of bla(KPC) genes in any meropenem-resistant Klebsiella spp. The PCR assay detected bla(KPC) in all of the reference strains, in 6 of 7 meropenem-resistant isolates, and in 0 of 62 meropenem-susceptible clinical isolates. The PCR assay was then used to confirm the presence of bla(KPC) in an additional 20 meropenem-resistant isolates from 16 patients. Restriction digestion of the PCR amplicons identified two bla(KPC) gene variants in our patient population: 9 isolates with C and 17 with T at nucleotide 944, consistent with bla(KPC-2) and bla(KPC-3), respectively. The real-time PCR assay is a rapid and accurate method to detect all KPC isoenzymes and was useful in documenting the presence and dissemination of KPC-producing strains in our patient population.


Considerations in physical therapy management of a non-responding patient with low back pain.
Case Study. Low back pain is a common condition managed by physical therapists (PT). Screening tools have been developed to assist the PT with medical screening of patients for serious disease. Sinister pathologies may present as musculoskeletal symptoms during the patient examination. It is important for the PT to frequently reevaluate their patient's response to therapeutic interventions and refer for further evaluation if they are not responding to conservative care. This case reports on the history and examination findings of a 36-year-old male presenting with recurrent low back pain. An emphasis is placed on the therapist's understanding and use of screening tools when interviewing patients and determining when medical referral may be indicated based on the patients history and examination findings. A review of the evidence on the diagnostic accuracy of screening strategies for malignancy in patients presenting with low back pain is presented. Sinister causes for low back pain are extremely rare. It is important that PTs be familiar with specific signs and symptoms that may indicate serious pathology when evaluating patients with low back pain. Identification of two red flags from this patient's history leads the clinician to refer the patient back to their primary care provider (PCP) for further investigation. They included: 1) the patient's inability to improve after one month and 2) clinicians' judgment. Use of "clinical judgment" may assist the PT in determining if their patient needs further investigation.


Celastrol inhibits colorectal cancer through TGF-β1/Smad signaling.
There are few clinical challenges associated with the treatment of colorectal cancer (CRC). Studies have shown that TGF-β plays a crucial role in CRC. Importantly, celastrol, a major components of the root extract of the traditional Chinese herb Tripterygium wilfordii Hook F, has been shown to inhibit the growth, adhesion, and metastasis of human CRC cells through the inhibition of TGF-β1/Smad signaling. Real-time PCR and Western blot tests were proceeded to present TGF-β1, TGF-β receptor type I (TGFβRI), TGF-β receptor type II (TGFβRII), Smad2/3, p-Smad2/3, Smad4, and glyceraldehyde-3-phosphate dehydrogenase expression in human colon cancer cell samples. Our results indicated that celastrol can reduce the expression levels of TGF-β1, TGFβRI, and TGFβRII in HCT116 and SW620 cells. Furthermore, celastrol could also prevent the increase in Smad4 and p-Smad2/3 in HCT116 and SW620 cells. Celastrol could inhibit tumor growth through TGF-β1/Smad signaling and might be a promising therapeutic component against CRC.


Ty1 sequence with enhancer and mating-type-dependent regulatory activities.
Some insertion mutations in Saccharomyces cerevisiae activate the expression of adjacent structural genes. The CYC7-H2 mutation is a Ty1 insertion 5' to the iso-2-cytochrome c coding region of CYC7. The Ty1 insertion causes a 20-fold increase in CYC7 expression in a and alpha haploid cell types of S. cerevisiae. This activation is repressed in the a/alpha diploid cell type. Previous computer analysis of the CYC7-H2 Ty1 activator region identified two related sequences with homology both to mammalian enhancers and to a yeast a/alpha control site. A 112-base-pair (bp) DNA fragment encompassing one of these blocks of homology functioned as one component of the Ty1 activator. A 28-bp synthetic oligonucleotide with the wild-type homology block sequence was also functional. A single base pair mutation within the enhancer core of the synthetic 28-bp regulatory element reduced its activation ability to near background amounts. In addition, the 112-bp Ty1 fragment by itself functioned as a target for repression of adjacent gene expression in a/alpha diploid cells.