Cigarette Smoking by Patients With Serious Mental Illness, 1999-2016: An Increasing Disparity.
This study examined the prevalence of cigarette smoking and the quantity of cigarettes consumed by individuals with schizophrenia and bipolar disorder and those without a psychiatric disorder in the period 1999-2016. A total of 1,938 individuals provided information about their cigarette smoking at enrollment into a research study for which they were selected without regard to their smoking status. Differences among groups and trends over time in smoking and cigarette consumption were examined by using multivariate models. Marked differences between groups were noted in the prevalence of smoking and in the quantity of cigarettes consumed. Overall, 62% of individuals with schizophrenia, 37% with bipolar disorder, and 17% of participants without a psychiatric disorder (control group) reported that they were current smokers. Smoking prevalence decreased over time in the sample primarily because of the decrease in smoking in the control group. Smokers with schizophrenia and with bipolar disorder smoked more cigarettes per day than smokers in the control group. Among smokers in all the groups, the quantity of cigarettes consumed per day declined significantly over the study period. Smoking was significantly associated with older age, less education, Caucasian race, and male gender. The prevalence of smoking has remained alarmingly high among individuals with schizophrenia and bipolar disorder, and the disparity with those without psychiatric disorders and with the general population is increasing. Additional measures are urgently needed to address this major public health problem.


[Computed tomography and magnetic resonance tomography of the normal temporal bone].
The normal anatomy of the temporal bone and the inner ear will be described in detail on high resolution computed tomography (HRCT) and magnetic resonance images. The imaging technique of computer tomography--either single detector or multi detector CT--is normally obtained in an axial plane without the intravenous application of contrast material. The images are reconstructed in a high resolution bone window level setting. The coronal images are reconstructed either if used single detector or multi detector CT. Only in some cases a scan in the coronal plane is directly obtained using a single detector CT. MR imaging of temporal bone is usually performed in a head coil. Axial high resolution 3D-T2-weighted sequences either in fast spin echo technique or gradient echo technique--for example CISS-sequence--are obtained, then an axial high resolution T1-weighted sequence before and after the application of gadopentate dimiglumine is performed. HRCT excellently demonstrates the osseous structures of the temporal bone as well as of the inner ear, while MRI excellently depicts soft tissue structures especially those of the inner ear. Due to the susceptibility artifacts MRI is not very suitable for imaging the external auditory canal or the middle ear or the pneumatic system. In conclusion HRCT is so far excellent to delineate the osseous structures of the temporal bone and inner ear while MRI excellently depicts the soft tissue structures of the inner ear, the internal auditory canal and the cerebellopontine angle. Reissner's membrane, the cochlear duct, and the organ of Corti cannot be visualized even using high-resolution MRI. HRCT and MRI are therefore used as complementary methods for imaging the temporal bone.


Design and analysis of ultra-compact EO polymer modulators based on hybrid plasmonic microring resonators.
Ultra-compact EO polymer modulators based on hybrid plasmonic microring resonators are proposed, simulated and analyzed. Comparing with Si slot microring modulator, hybrid plasmonic microring modulator shows about 6-times enhancement of the figure of merit when the bending radius is around 510 nm, due to its much larger intrinsic quality factor in sub-micron radius range. Influences of the EO polymer height and Si height on the device's performance are analyzed and optimal design is given. When operating with a bias of 3.6 V, the proposed device has optical modulation amplitude of 0.8 and insertion loss of about 1 dB. The estimated power consumption is about 5 fJ/bit at 100 GHz.


Bimolecular affinity purification: a variation of TAP with multiple applications.
The identification of true interacting partners of any given bait can be plagued by the nonspecific purification of irrelevant proteins. To avoid this problem, Tandem Affinity Purification (TAP) is a widely used procedure in molecular biology as this reduces the chance of nonspecific proteins being present in the final preparation. In this approach, two different affinity tags are fused to the protein bait. Herein, we review in detail a variation on the TAP procedure that we have previously developed, where the affinity moieties are placed on two different proteins that form a complex in vivo. This variation, which we refer to as Bimolecular Affinity Purification (BAP), is suited for the identification of specific molecular complexes marked by the presence of two known proteins. We have utilized BAP for characterization of molecular complexes and evaluation of proteins interaction. Another application of BAP is the isolation of ubiquitin-like proteins (UBL)-modified fractions of a given protein and characterization of the lysine-acceptor site and structure of UBL-chains.


Comparison of oral and vaginal metronidazole for treatment of bacterial vaginosis in pregnancy: impact on fastidious bacteria.
Bacterial vaginosis (BV) is a common condition that is associated with preterm birth and acquisition of complex communities of vaginal bacteria that include several fastidious species. Treatment of BV in pregnancy has mixed effects on the risk of preterm delivery, which some hypothesize is due to variable antibiotic efficacy for the fastidious bacteria. Both oral and intravaginal metronidazole can be used to treat bacterial vaginosis in pregnancy, but little is known about the impact of different routes of antibiotic administration on concentrations of fastidious vaginal bacteria. This was a sub-study of a larger randomized trial of oral versus vaginal metronidazole for treatment of BV in pregnancy. Fifty-three women were evaluated, including 30 women who received oral metronidazole and 23 who received intravaginal metronidazole. Bacterial taxon-specific quantitative PCR assays were used to measure concentrations of bacterial vaginosis associated bacterium (BVAB) 1, 2, and 3, Gardnerella vaginalis, Atopobium species, Leptotrichia/Sneathia species, Megasphaera species, and Lactobacillus crispatus before and after antibiotic treatment. Concentrations of Leptotrichia and Sneathia spp. and the fastidious Clostridia-like bacterium designated BVAB1 decreased significantly with oral (p = .002, p = .02) but not vaginal therapy (p = .141, p = .126). The fastidious bacterium BVAB3 did not significantly decrease with either treatment. Concentrations of Atopobium spp., reportedly resistant to metronidazole in vitro, dropped significantly with oral (p = .002) and vaginal (p = .001) treatment. There was no significant difference in the magnitude of change in bacterial concentrations between oral and vaginal treatment arms for any of the bacterial species. Lactobacillus crispatus concentrations did not change. Both oral and vaginal metronidazole therapy in pregnant women result in a significant decrease in concentrations of most BV-associated anaerobic bacteria, with the exception that Leptotrichia, Sneathia and BVAB1 do not significantly decrease with vaginal metronidazole therapy. These data suggest that the route of antibiotic administration has a minor impact on bacterial eradication in pregnant women with BV. This trial is registered with ClinicalTrials.gov, number NCT00153517.


Genome-wide gene order distances support clustering the gram-positive bacteria.
Initially using 143 genomes, we developed a method for calculating the pair-wise distance between prokaryotic genomes using a Monte Carlo method to estimate the conservation of gene order. The method was based on repeatedly selecting five or six non-adjacent random orthologs from each of two genomes and determining if the chosen orthologs were in the same order. The raw distances were then corrected for gene order convergence using an adaptation of the Jukes-Cantor model, as well as using the common distance correction D' = -ln(1-D). First, we compared the distances found via the order of six orthologs to distances found based on ortholog gene content and small subunit rRNA sequences. The Jukes-Cantor gene order distances are reasonably well correlated with the divergence of rRNA (R (2) = 0.24), especially at rRNA Jukes-Cantor distances of less than 0.2 (R (2) = 0.52). Gene content is only weakly correlated with rRNA divergence (R (2) = 0.04) over all distances, however, it is especially strongly correlated at rRNA Jukes-Cantor distances of less than 0.1 (R (2) = 0.67). This initial work suggests that gene order may be useful in conjunction with other methods to help understand the relatedness of genomes. Using the gene order distances in 143 genomes, the relations of prokaryotes were studied using neighbor joining and agreement subtrees. We then repeated our study of the relations of prokaryotes using gene order in 172 complete genomes better representing a wider-diversity of prokaryotes. Consistently, our trees show the Actinobacteria as a sister group to the bulk of the Firmicutes. In fact, the robustness of gene order support was found to be considerably greater for uniting these two phyla than for uniting any of the proteobacterial classes together. The results are supportive of the idea that Actinobacteria and Firmicutes are closely related, which in turn implies a single origin for the gram-positive cell.


Clarin-1 expression in adult mouse and human retina highlights a role of Müller glia in Usher syndrome.
Usher syndrome type 3 (USH3) is an autosomal recessively inherited disorder caused by mutations in the gene clarin-1 (CLRN1), leading to combined progressive hearing loss and retinal degeneration. The cellular distribution of CLRN1 in the retina remains uncertain, either because its expression levels are low or because its epitopes are masked. Indeed, in the adult mouse retina, Clrn1 mRNA is developmentally downregulated, detectable only by RT-PCR. In this study we used the highly sensitive RNAscope in situ hybridization assay and single-cell RNA-sequencing techniques to investigate the distribution of Clrn1 and CLRN1 in mouse and human retina, respectively. We found that Clrn1 transcripts in mouse tissue are localized to the inner retina during postnatal development and in adult stages. The pattern of Clrn1 mRNA cellular expression is similar in both mouse and human adult retina, with CLRN1 transcripts being localized in Müller glia, and not photoreceptors. We generated a novel knock-in mouse with a hemagglutinin (HA) epitope-tagged CLRN1 and showed that CLRN1 is expressed continuously at the protein level in the retina. Following enzymatic deglycosylation and immunoblotting analysis, we detected a single CLRN1-specific protein band in homogenates of mouse and human retina, consistent in size with the main CLRN1 isoform. Taken together, our results implicate Müller glia in USH3 pathology, placing this cell type to the center of future mechanistic and therapeutic studies to prevent vision loss in this disease. © 2019 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.


Efficacy of a broad host range lytic bacteriophage against E. coli adhered to urothelium.
Persistent urinary tract infections (UTI) are often caused by E. coli adhered to urothelium. This type of cells is generally recognized as very tolerant to antibiotics which renders difficult the treatment of chronic UTI. This study investigates the use of lytic bacteriophages as alternative antimicrobial agents, particularly the interaction of phages with E. coli adhered to urothelium and specifically determines their efficiency against this type of cells. The bacterial adhesion to urothelium was performed varying the bacterial cell concentrations and the period and conditions (static, shaken) of adhesion. Three collection bacteriophages (T1, T4, and phiX174 like phages) were tested against clinical E. coli isolates and only one was selected for further infection experiments. Based on the lytic spectrum against clinical isolates and its ability to infect the highest number of antibiotic resistant strains, the T1-like bacteriophage was selected. This bacteriophage caused nearly a 45% reduction of the bacterial population after 2 h of treatment. This study provides evidence that bacteriophages are effective in controlling suspended and adhered cells and therefore can be a viable alternative to antibiotics to control urothelium- adhered bacteria.


New advances in fertility preservation for pediatric cancer patients.
The number of pediatric cancer survivors is growing rapidly as treatments become more effective. However, many current regimens cause gonadotoxicity and permanent infertility, significantly impacting quality of life. The purpose of this review is to update pediatric oncologists on risk factors for cancer treatment-associated gonadotoxicity, current methods for fertility preservation, and new scientific advances in this area. Infertility is an enormous quality-of-life issue for pediatric cancer survivors and their families. Numerous treatment options are already available to prevent infertility in patients at risk. It is important to counsel patients at risk and initiate management for fertility preservation prior to beginning therapy. Preclinical research indicates that it may be possible to bank gonadal tissues from patients for subsequent re-implantation after therapy or expansion of germ cells in vitro. Further translational studies are required to advance these technologies into clinical use. Pediatric cancer survivors are at risk for long-term treatment-related gonadal failure and infertility. Counseling and treatment should begin prior to initiating chemo or radiation therapy. Recent scientific advances in understanding germ cell biology should eventually generate new clinical strategies to maintain fertility in pediatric cancer patients.


Transforming Growth Factor-β Promotes Liver Tumorigenesis in Mice via Up-regulation of Snail.
Transforming growth factor beta (TGF-β) suppresses early stages of tumorigenesis, but also contributes to migration and metastasis of cancer cells. A large number of human tumors contain mutations that inactivate its receptors, or downstream proteins such as Smad transcription factors, indicating that the TGF-β signaling pathway prevents tumor growth. We investigated the effects of TGF-β inhibition on liver tumorigenesis in mice. C57BL/6 mice received hydrodynamic tail-vein injections of transposons encoding HRASG12V and a short hairpin RNA (shRNA) to down-regulate p53, or those encoding HRASG12V and MYC, or those encoding HRASG12V and TAZS89A, to induce liver tumor formation; mice were also given injections of transposons encoding SMAD7 or shRNA against SMAD2, SMAD3, SMAD4, or SNAI1 (Snail), with or without ectopic expression of Snail. Survival times were compared, and livers were weighted and examined for tumors. Liver tumor tissues were analyzed by quantitative reverse-transcription PCR, RNA sequencing, immunoblots, and immunohistochemistry. We analyzed gene expression levels in human hepatocellular carcinoma samples deposited in The Cancer Genome Atlas. A cell proliferation assay was performed using human liver cancer cell lines (HepG2 and Huh7) stably expressing Snail or shRNA against Snail. TGF-β inhibition via overexpression of SMAD7 (or knockdown of SMAD2, SMAD3, or SMAD4) consistently reduced formation and growth of liver tumors in mice that expressed activated RAS plus shRNA against p53, or in mice that expressed activated RAS and TAZ. TGF-β signaling activated transcription of the Snail gene in liver tumors induced by HRASG12V and shRNA against p53, and by activated RAS and TAZ. Knockdown of Snail reduced liver tumor formation in both tumor models. Ectopic expression of Snail restored liver tumorigenesis suppressed by disruption of TGF-β signaling. In human hepatocellular carcinoma, Snail expression correlated with TGF-β activation. Ectopic expression of Snail increased cellular proliferation, whereas Snail knockdown led to reduced proliferation in human hepatocellular carcinoma cells. In analyses of transgenic mice, we found TGF-β signaling to be required for formation of liver tumors upon expression of activated RAS and shRNA down-regulating p53, and upon expression of activated RAS and TAZ. Snail is the TGF-β target that is required for hepatic tumorigenesis in these models.


Curving out a new path: CD28/B7 cis interactions.
Important signaling events at the immunological synapse have increasingly been linked to cis interactions between receptors on T cells. In this issue of Immunity, Zhao et al.1 implicate cis CD28/B7 interactions facilitated by curved membrane invaginations in boosting tumor immunity.


Assessing large spatial scale landscape change effects on water quality and quantity response in the lower Athabasca River basin.
Increased land use intensity has been shown to adversely affect aquatic ecosystems. Multiple landscape stressors interact over space and time, producing cumulative effects. Cumulative Effects Assessment (CEA) is the process of evaluating the impact a development project may have on the ecological surroundings, but several challenges exist that make current approaches to cumulative effects assessment ineffective. The main objective of this study was to compare results of different methods used to link landscape stressors with stream responses in a highly developed watershed, where past work has shown that the river has experienced significant water quality and quantity changes to improve approaches to CEA. The study site was the lower reaches of the Athabasca River, Canada that have been subjected to a diverse range of intense anthropogenic developments since the late 1960s. Linkages between landscape change and river response were evaluated using correlation analyses, stepwise, multiple regression, and regression trees. Notable landscape changes include increased industrial development and forest cut-blocks, made evident from satellite imagery and supporting ancillary data sets. Simple regression analyses showed water use was closely associated with total phosphorus (TP) and Na(+) concentrations, as well as specific conductance. The regression trees for total organic carbon (TOC), TP, and Na(+) showed that the landscape variables that appear as the first characteristic were the same variables that showed significant relations for their respective simple regression models. Simple, stepwise, and multiple regressions in conjunction with regression trees were useful in this study for capturing the strongest associations between landscape stressors and river response variables. The results highlight the need for improved scaling methods and monitoring strategies crucial to managing cumulative effects to river systems.


PRPF4 mutations cause autosomal dominant retinitis pigmentosa.
Retinitis pigmentosa (RP), a disease characterized by progressive loss of photoreceptors, exhibits significant genetic heterogeneity. Several genes associated with U4/U6-U5 triple small nuclear ribonucleoprotein (tri-snRNP) complex of the spliceosome have been implicated in autosomal dominant RP (adRP). HPrp4, encoded by PRPF4, regulates the stability of U4/U6 di-snRNP, which is essential for continuous splicing. Here, we identified two heterozygous variants in PRPF4, including c.-114_-97del in a simplex RP patient and c.C944T (p.Pro315Leu), which co-segregates with disease phenotype in a family with adRP. Both variants were absent in 400 unrelated controls. The c.-114_-97del, predicted to affect two transcription factor binding sites, was shown to down-regulate the promoter activity of PRPF4 by a luciferase assay, and was associated with a significant reduction of PRPF4 expression in the blood cells of the patient. In fibroblasts from an affected individual with the p.Pro315Leu variant, the expression levels of several tri-snRNP components, including PRPF4 itself, were up-regulated, with altered expression pattern of SC35, a spliceosome marker. The same alterations were also observed in cells over expressing hPrp4(Pro315Leu), suggesting that they arose as a compensatory response to a compromised splicing mechanism caused by hPrp4 dysfunction. Further, over expression of hPrp4(Pro315Leu), but not hPrp4(WT), triggered systemic deformities in wild-type zebrafish embryos with the retina primarily affected, and dramatically augmented death rates in morphant embryos, in which orthologous zebrafish prpf4 gene was silenced. We conclude that mutations of PRPF4 cause RP via haploinsufficiency and dominant-negative effects, and establish PRPF4 as a new U4/U6-U5 snRNP component associated with adRP.


Hemodynamic segmentation of MR brain perfusion images using independent component analysis, thresholding, and Bayesian estimation.
Dynamic-susceptibility-contrast MR perfusion imaging is a widely used imaging tool for in vivo study of cerebral blood perfusion. However, visualization of different hemodynamic compartments is less investigated. In this work, independent component analysis, thresholding, and Bayesian estimation were used to concurrently segment different tissues, i.e., artery, gray matter, white matter, vein and sinus, choroid plexus, and cerebral spinal fluid, with corresponding signal-time curves on perfusion images of five normal volunteers. Based on the spatiotemporal hemodynamics, sequential passages and microcirculation of contrast-agent particles in these tissues were decomposed and analyzed. Late and multiphasic perfusion, indicating the presence of contrast agents, was observed in the choroid plexus and the cerebral spinal fluid. An arterial input function was modeled using the concentration-time curve of the arterial area on the same slice, rather than remote slices, for the deconvolution calculation of relative cerebral blood flow.


Randomised controlled trial of general practitioner intervention in patients with excessive alcohol consumption.
To determine effectiveness of advice from general practitioners to heavy drinkers to reduce their excessive alcohol consumption (35 U or more a week for men, 21 U or more for women). Randomised, controlled double blind trial over 12 months with interim assessment at six months. Group practices (n = 47; list size averaging 10,000) recruited from Medical Research Council's general practice research framework, mostly in rural or small urban settings. Patients recruited after questionnaire survey. Of total of 2571 (61.2%) of 4203 patients invited for interview who attended, 909 (35.4%) stated that in past seven days they had drunk above the limits set for study and had not received advice; they were randomised to control and treatment groups. Patients in treatment group were interviewed by general practitioner (who had had a training session) and received advice and information about how to reduce consumption and also given a drinking diary. Study aimed at detecting a reduction in proportion of men with excessive alcohol consumption of 30% in treatment group and 20% in control group (for women 40% and 20%, respectively) with a power of 90% at 5% level of significance. In addition, corroborative measures such as estimation of gamma-glutamyltransferase activity were included. At one year a mean reduction in consumption of alcohol of 18.2 (SE 1.5) U/week had occurred in treated men compared with a reduction of 8.1 (1.6) U/week in controls (p less than 0.001). The proportion of men with excessive consumption at interview had dropped by 43.7% in the treatment group compared with 25.5% in controls (p less than 0.001). A mean reduction in weekly consumption of 11.5 (1.6) U occurred in treated women compared with 6.3 (2.0) U in controls (p less than 0.05), with proportionate reductions of excessive drinkers in treatment and control groups of 47.7% and 29.2% respectively. Reduction in consumption increased significantly with number of general practitioner interventions. At one year the mean value for gamma-glutamyltransferase activity had dropped significantly more in treated men (-2.4 (0.9)IU/l) than in controls (+1.1(1.0)IU/l; t = 2.7, p less than 0.01). Reduction in gamma-glutamyltransferase activity tended to increase with number of intervention sessions in men. Changes in gamma-glutamyltransferase activity in women and changes in other indicators in both sexes did not differ significantly between treatment and control groups. If the results of this study were applied to the United Kingdom intervention by general practitioners could each year reduce to moderate levels the alcohol consumption of some 250000 men and 67500 women who currently drink to excess. General practitioners and other members of the primary health care team should therefore be encouraged to include counselling about alcohol consumption in their preventive activities.


Next Speakers Plan Their Turn Early and Speak after Turn-Final "Go-Signals".
In conversation, turn-taking is usually fluid, with next speakers taking their turn right after the end of the previous turn. Most, but not all, previous studies show that next speakers start to plan their turn early, if possible already during the incoming turn. The present study makes use of the list-completion paradigm (Barthel et al., 2016), analyzing speech onset latencies and eye-movements of participants in a task-oriented dialogue with a confederate. The measures are used to disentangle the contributions to the timing of turn-taking of early planning of content on the one hand and initiation of articulation as a reaction to the upcoming turn-end on the other hand. Participants named objects visible on their computer screen in response to utterances that did, or did not, contain lexical and prosodic cues to the end of the incoming turn. In the presence of an early lexical cue, participants showed earlier gaze shifts toward the target objects and responded faster than in its absence, whereas the presence of a late intonational cue only led to faster response times and did not affect the timing of participants' eye movements. The results show that with a combination of eye-movement and turn-transition time measures it is possible to tease apart the effects of early planning and response initiation on turn timing. They are consistent with models of turn-taking that assume that next speakers (a) start planning their response as soon as the incoming turn's message can be understood and (b) monitor the incoming turn for cues to turn-completion so as to initiate their response when turn-transition becomes relevant.


Genetic features of blaNDM-1-positive Enterobacteriaceae.
Genetic features associated with the bla(NDM-1) gene were investigated in 6 Escherichia coli, 7 Klebsiella pneumoniae, 1 Citrobacter freundii, 1 Proteus mirabilis, and 1 Providencia stuartii isolate of worldwide origin. Clonal diversity was observed for both E. coli and K. pneumoniae. The bla(NDM-1) gene was carried by different plasmid types (IncA/C, IncF, IncL/M, or untypeable) and was likely chromosome borne in two isolates. The bla(NDM-1) plasmids coharbored a variety of resistance determinants, including β-lactamase genes, quinolone resistance genes, and 16S RNA methylase genes.


Transcriptional control of expression of genes for photosynthetic reaction center and light-harvesting proteins in the purple bacterium Rhodovulum sulfidophilum.
The purple photosynthetic bacterium Rhodovulum sulfidophilum synthesizes photosynthetic apparatus even under highly aerated conditions in the dark. To understand the oxygen-independent expression of photosynthetic genes, the expression of the puf operon coding for the light-harvesting 1 and reaction center proteins was analyzed. Northern blot hybridization analysis showed that puf mRNA synthesis was not significantly repressed by oxygen in this bacterium. High-resolution 5' mapping of the puf mRNA transcriptional initiation sites and DNA sequence analysis of the puf upstream regulatory region indicated that there are three possible promoters for the puf operon expression, two of which have a high degree of sequence similarity with those of Rhodobacter capsulatus, which shows a high level of oxygen repression of photosystem synthesis. Deletion analysis showed that the third promoter is oxygen independent, but the activity of this promoter was not enough to explain the aerobic level of mRNA. The posttranscriptional puf mRNA degradation is not significantly influenced by oxygen in R. sulfidophilum. From these results, we conclude that puf operon expression in R. sulfidophilum is weakly repressed by oxygen, perhaps as a result of the following: (i) there are three promoters for puf operon transcription, at least one of which is oxygen independent; (ii) readthrough transcripts which may not be affected by oxygen may be significant in maintaining the puf mRNA levels; and (iii) the puf mRNA is fairly stable even under aerobic conditions.


Critical elements of the immunoglobulin heavy chain gene enhancers for deregulated expression of bcl-2.
Translocation of the bcl-2 gene to the immunoglobulin heavy chain gene is the most common alteration in follicular lymphoma. The result is the deregulated expression of bcl-2 and increased resistance to cell death. Regulation of the immunoglobulin heavy chain gene is controlled in part by four DNase I-hypersensitive regions located 3' of the gene. Here, we show that these four enhancer regions also contribute to bcl-2 up-regulation in t(14;18) cells. The enhancers are able to individually or in combination activate bcl-2 promoter activity. The HS4 enhancer region was found to impart the largest positive effect on the bcl-2 promoter, activating it by 6-fold, whereas addition of the HS1,2 region with HS4 increased promoter activity by approximately 9-fold. Nuclear factor kappaB binding sites were shown to be primarily responsible for the positive activity contributed by the HS1,2 and HS4 regions, and we observed the in vivo interaction of these factors with the human immunoglobulin heavy chain gene enhancer regions in t(14;18) cells. In addition, two Sp1 binding sites in HS4 were also found to positively influence bcl-2 activity, and Sp1 was observed to interact with the human HS4 enhancer in vivo. These results suggest that the interactions of the nuclear factor kappaB and Sp1 transcription factors with the immunoglobulin heavy chain enhancer region are important for bcl-2 deregulation in t(14;18) cells.


A double-blind trial of zinc supplementation in pregnancy.
A double-blind randomised trial of oral zinc supplementation was carried out during the last two trimesters of pregnancy. Fifty-six women at risk of delivering a small-for-gestational-age baby received either zinc supplement (22.5 mg daily) or placebo. Twenty-nine of the women were compliant. Zinc significantly reduced the incidence of intrauterine growth retardation, and most measured indices of labour and fetal health were better in the supplemented group. Larger studies are now needed to confirm a role for selective zinc supplementation in at-risk pregnancies.


Association of Life's Essential 8 With Cardiovascular Disease and Mortality: The Framingham Heart Study.
The association of the American Heart Association's updated cardiovascular health score, the Life's Essential 8 (LE8), with cardiovascular disease (CVD) and death is not described in the FHS (Framingham Heart Study). We evaluated Framingham Offspring participants at examinations 2 and 6 (n=2888 and 1667; and mean age, 44 and 57 years, respectively), free of CVD with information on LE8 components. Using age-sex-adjusted Cox models, we related LE8 and its change (examination 2 to examination 6) with CVD and death risk and compared associations with those of the Life's Simple 7 score. Mean LE8 score at examination 2 was 67 points (minimum, 26 points; maximum, 100 points). At both examinations, participants were reclassified to a different cardiovascular health status, depending on which method (LE8 versus Life's Simple 7) was used (60% of participants in ideal Life's Simple 7 status were in intermediate LE8 category). On follow-up after examination 2 (median, 30 and 33 years for CVD and death, respectively), we observed 966 CVD events, and 1195 participants died. Participants having LE8≥68 (sample median) were at lower CVD and death risk compared with those with LE8<68 (examination 2: CVD hazard ratio [HR], 0.47 [95% CI, 0.41-0.54]; death HR, 0.55 [95% CI, 0.49-0.62]; all P<0.001). Participants maintaining low LE8 scores during life course were at highest CVD and death risk (CVD: HRs ranging from 1.8 to 2.3; P<0.001; death HR, 1.45 [95% CI, 1.13-1.85]; P=0.003 versus high-high group). Further studies are warranted to elucidate whether the LE8 score is a better marker of CVD and death risk, compared with Life's Simple 7 score.


Quasi-elastic light-scattering studies of aqueous biliary lipid systems. Cholesterol solubilization and precipitation in model bile solutions.
We have employed quasi-elastic light-scattering methods to characterize micellar aggregates and microprecipitates formed in aqueous solutions containing sodium taurocholate (TC), egg lecithin (L), and cholesterol (Ch). Particle size and polydispersity were studied as functions of Ch mole fraction (XCh = 0-15%), L/TC molar ratio (0-1.6), temperature (5-85 degrees C), and total lipid concentration (3 and 10 g/dL in 0.15 M NaCl). For XCh values below the established solubilization limits (XChmax) [Carey, M. C., & Small, D. M. (1978) J. Clin. Invest. 61, 998], added Ch has little influence on the size of simple TC micelles (type 1 systems), on the coexistence of simple and mixed TC-L micelles (type 2 systems), or on the growth of "mixed disc" TC-L micelles (type 3 systems). For supersaturated systems (XCh/XChmax greater than 1), 10 g/dL type 1 systems (L/TC = 0) exist as metastable micellar solutions even at XCh/XChmax = 5.3. Metastability is decreased in type 2 systems (0 less than L/TC less than 0.6), and "labile" microprecipitation occurs when XCh/XChmax exceeds approximately 1.6. In 10 g/dL mixtures, the microprecipitates initially range in size from 500 to 20000 A and later coalesce to form a buoyant macroscopic precipitate phase. In 3 g/dL mixtures, the microprecipitates are smaller (200-400 A) and remain as a stable, noncoalesced microdispersion. Transmission electron microscopy of the microprecipitates formed at both concentrations indicates a globular noncrystalline structure, and lipid analysis reveals the presence of cholesterol and lecithin in a molar ratio (Ch/L) of approximately 2/1, suggesting that the microprecipitates represent a metastable cholesterol-rich liquid-crystalline phase. In supersaturated type 3 systems (0.6 less than L/TC less than 2.0), the precipitate phase is a lecithin-rich liquid-crystalline phase which likewise coalesces in a 10 g/dL system but forms stable vesicle (liposomal) structures (600-800 A radius) in 3 g/dL systems. In conjunction with these experimental data, we present a quantitative thermodynamic analysis of Ch solubilization in model bile systems from which rigorous deductions of the free energy and enthalpy change for solubilization of cholesterol monohydrate in type 1 and type 2 systems are obtained. In addition, we employ homogeneous nucleation theory to analyze the origin of the metastable/labile limit in supersaturated systems and to deduce the interfacial tension between microprecipitates and solution. On the basis of these experimental data and theoretical analyses, we offer new hypotheses on the structure and physiology of bile and the pathogenesis of Ch gallstones. In particular, it is suggested that the "stable" microprecipitates observed in 3 g/dL type 2 systems may provide a secondary vehicle (in addition to micelles) for cholesterol transport in supersaturated hepatic bile.


[The effectiveness of using biohumus for growing plants under autonomous conditions].
There has been investigated the possibility of prolonging the useful time of the substrate on the basis of natural salt-loaded zeolite "Balkanin" used in space greenhouses to grow plants through introduction of an organic addition, i.e. biohumus, instead of regeneration. Biohumus has been applied into the substrate in amounts of 5.0-15.0 wt%. There has been grown tropical perennial plant Basella rubra L. used for food as a salad. It is found that the periodic introduction of biohumus in substrate enables a 4-19 time increase of its useful life without regeneration within more than 4 years and assures high and stable crop yield with high quality biomass. As compared to the control, the nitrates content of the plant biomass was, on the average, 3 times lower and decreased to the point of their total absence at the end of experiment. The results obtained allow us to recommend the above-mentioned technology of plant growing in space greenhouses both when included into closed environmental systems and in independent plant growth facilities aboard spacecraft. During space flights exceeding 2 years in length, additional supplies of biohumus will be required. This will be enough to stabilize the structure and the size of microbial community in substrate and provide a high crop yield.


The implementation of prolonged exposure: Design of a multisite study evaluating the usefulness of workshop with and without consultation.
This randomized trial examines the dissemination and implementation of prolonged exposure (PE) therapy for posttraumatic stress symptoms in U.S. Army medical treatment facilities. The study compares two PE training models: Standard PE training, comprised of a 4-day workshop only, and Extended PE training, comprised of a 4-day workshop plus expert case consultation. Behavioral health providers (N=180) across three medium-to-large Army installations will be randomly assigned to either Standard PE training or Extended PE training. Changes in provider attitudes will be examined across groups. After completing PE training, the use of PE components with patients reporting posttraumatic stress symptoms and clinical outcomes of these participating patients (N=500) will be examined. This article describes the rationale and methods of the study. In addition, a number of methodological issues in conducting a multisite naturalistic study in the U.S. Army are discussed.


ChIP-seq accurately predicts tissue-specific activity of enhancers.
A major yet unresolved quest in decoding the human genome is the identification of the regulatory sequences that control the spatial and temporal expression of genes. Distant-acting transcriptional enhancers are particularly challenging to uncover because they are scattered among the vast non-coding portion of the genome. Evolutionary sequence constraint can facilitate the discovery of enhancers, but fails to predict when and where they are active in vivo. Here we present the results of chromatin immunoprecipitation with the enhancer-associated protein p300 followed by massively parallel sequencing, and map several thousand in vivo binding sites of p300 in mouse embryonic forebrain, midbrain and limb tissue. We tested 86 of these sequences in a transgenic mouse assay, which in nearly all cases demonstrated reproducible enhancer activity in the tissues that were predicted by p300 binding. Our results indicate that in vivo mapping of p300 binding is a highly accurate means for identifying enhancers and their associated activities, and suggest that such data sets will be useful to study the role of tissue-specific enhancers in human biology and disease on a genome-wide scale.


Electrochemical sensing of concanavalin A and ovalbumin interaction in solution.
In an attempt to develop a label- and reagent-free electrochemical method for the detection of lectin-glycoprotein interactions, we tested lectin-concanavalin A (ConA), glycoprotein-ovalbumin (Ova) and their complex using chronopotentiometric stripping (CPS) analysis and a hanging mercury drop electrode. Incubation of ConA with Ova resulted in an increase of the CPS peak H of the complex as compared to the CPS peaks of individual Ova and ConA proteins. Qualitatively similar results were obtained with other glycoprotein-lectin couples (ConA-RNase B and lectin from Sambucus nigra-fetuin). Using the CPS method, we were able to follow the course of complex formation in solution. Comparable responses of Ova, ConA and ConA-Ova complex were obtained not only at the mercury electrode but also with solid amalgam electrodes, which are more suitable for parallel analysis. It can be anticipated that electrochemical methods, namely CPS, will find application in glycomics and proteomics.


[Remote results of aortofemoral reconstructions performed through miniaccess].
Analysed herein are long-term outcomes of 250 reconstructions of the infrarenal aorta for occlusive disease with the use of transperitoneal miniapproach (median minilaparotomy 5-10 cm long). The patients' average age amounted to 58.2±0.64 years. 60.4% of patients had critical ischaemia. Bilateral reconstruction was performed in 216 patients, and unilateral--in 34 subjects (with revascularization performed on a total of 466 limbs). Hospital lethality rate amounted to 0.8% (2 patients). There were 2 bypass-graft thromboses (in one case with successful thrombectomy). The incidence rate of amputations was 0.86% from the number of revascularized extremities. Remote outcomes were assessed in 145 patients within terms varying from 6 months to 84 months (mean follow-up duration--30.8±1.4 months). Primary patency after 1 year amounted to 97.0±0.1%, after 5 years--81.8±3.9%, after 84 months--67. ±9.8%, with the corresponding figures for secondary patency amounting to 98.0±0.83%, 91.9±2.8% and 91.9±2.8%, respectively. The limb preservation rate after one year equalled 97.0±0.9%, after 5 years--93.6±2.4%, after 84 months--93.6±2.4%. Five-year survival amounted to 73.5±6.2%. Postoperative ventral hernias were revealed in 6.9% of patients. Hence, the long-term outcomes of aortofemoral reconstructions thorough the miniapproach correspond to the commonly accepted criteria for quality of operations for occlusive pathology of the aortofemoral segment.


Progress toward poliomyelitis eradication--Ethiopia, 1997-August 2000.
In 1988, the World Health Assembly resolved to eradicate poliomyelitis globally by 2000 (1). Following the signing of the Yaounde Declaration on Polio Eradication in Africa in 1996, Ethiopia joined global efforts toward polio eradication (2). Since then, Ethiopia has accelerated implementation of polio eradication strategies. This report summarizes progress toward polio eradication in Ethiopia during 1997-August 2000 and highlights the remaining challenges toward achieving the goal.


Synthetic activities of cultured retinal pigment epithelial cells from a patient with retinitis pigmentosa.
We established cultures of retinal pigment epithelial (RPE) cells from four normal individuals and a 56-year-old male patient with autosomal-dominant retinitis pigmentosa (RP) and examined the synthetic activities of these cells via radiolabeling experiments. The uptake of radioactive precursors per milligram of cell protein by RPE cells from the patient with RP was significantly higher than that found in the normal control cells. It appears that the net synthetic activities of glycosaminoglycans and proteins in our patient's cells were enhanced and that the cell metabolism was altered. This finding suggests that malfunctioning of the RPE cells may play a role in the deterioration of photoreceptor functions seen in at least some patients with autosomal-dominant RP.


Cytoplasmic pH response to acid stress in individual cells of Escherichia coli and Bacillus subtilis observed by fluorescence ratio imaging microscopy.
The ability of Escherichia coli and Bacillus subtilis to regulate their cytoplasmic pH is well studied in cell suspensions but is poorly understood in individual adherent cells and biofilms. We observed the cytoplasmic pH of individual cells using ratiometric pHluorin. A standard curve equating the fluorescence ratio with pH was obtained by perfusion at a range of external pH 5.0 to 9.0, with uncouplers that collapse the transmembrane pH difference. Adherent cells were acid stressed by switching the perfusion medium from pH 7.5 to pH 5.5. The E. coli cytoplasmic pH fell to a value that varied among individual cells (range of pH 6.2 to 6.8), but a majority of cells recovered (to pH 7.0 to 7.5) within 2 min. In an E. coli biofilm, cells shifted from pH 7.5 to pH 5.5 failed to recover cytoplasmic pH. Following a smaller shift (from pH 7.5 to pH 6.0), most biofilm cells recovered fully, although the pH decreased further than that of isolated adherent cells, and recovery took longer (7 min or longer). Some biofilm cells began to recover pH and then failed, a response not seen in isolated cells. B. subtilis cells were acid shifted from pH 7.5 to pH 6.0. In B. subtilis, unlike the case with E. coli, cytoplasmic pH showed no "overshoot" but fell to a level that was maintained. This level of cytoplasmic pH post-acid shift varied among individual B. subtilis cells (range of pH, 7.0 to 7.7). Overall, the cytoplasmic pHs of individual bacteria show important variation in the acid stress response, including novel responses in biofilms.


Indentation Strengths of Zirconium Diboride: Intrinsic versus Extrinsic Mechanisms.
Zirconium diboride (ZrB2) is an important ultra-high-temperature ceramic, which exhibits outstanding mechanical properties and is widely used in extreme environments. Extensive experimental studies, however, have found that synthesized ZrB2 specimens show widely scattered indentation hardness values ranging from 8.7 to 26 GPa. We have performed comprehensive stress-strain calculations of ZrB2 to explore its structural and stress responses and found that ZrB2 possesses an intrinsic indentation strength of 32.7 GPa, which is on par with those of other transition-metal borides that exhibit higher indentation hardness values of ∼30 GPa. This result suggests that large variations in measured hardness are driven by extrinsic factors, and an analysis of available experimental data indicates that the quality of the crystallinity of specimens holds the key to realizing improved hardness corresponding to the predicted intrinsic indentation strength. These findings offer insights into the origin of the previously reported lower hardness values of ZrB2 and raise the prospects of achieving superior strengths in well-crystallized ZrB2 that approach or match those of other ultrahard transition-metal compounds.


Assessment of myocardial perfusion by contrast echocardiography.
Contrast echocardiography delineates myocardial perfusion and has the potential for quantitating coronary flow and assessing myocardial viability. These applications add important physiologic information to the anatomic information readily available from conventional echocardiography. Because it can be rapidly performed at the bedside, contrast echocardiography may be valuable tool for the use in acute myocardial ischemia. When contrast echocardiography has been used after recanalization of occluded coronary arteries, the assessment of myocardial salvage conveys information concerning reflow, stunning, and prognosis, and in the case of angioplasty it provides immediate information regarding the success of the procedure. Contrast echocardiography can also assess myocardial areas at risk of irreversible damage and the presence or absence of collateral flow. Myocardial contrast echocardiography is a rapidly changing field and with the continued development of newer contrast agents and refinement of ultrasound imaging equipment, the applications of contrast echocardiography will continue to grow.


An obsolete dichotomy? Rethinking the rural–urban interface in terms of food security and production in the global south.
The global food system is coming under increasing strain in the face of urban population growth. The recent spike in global food prices (2007–08) provoked consumer protests, and raised questions about food sovereignty and how and where food will be produced. Concurrently, for the first time in history the majority of the global population is urban, with the bulk of urban growth occurring in smaller-tiered cities and urban peripheries, or 'peri-urban' areas of the developing world. This paper discusses the new emerging spaces that incorporate a mosaic of urban and rural worlds, and reviews the implications of these spaces for livelihoods and food security. We propose a modified livelihoods framework to evaluate the contexts in which food production persists within broader processes of landscape and livelihood transformation in peri-urban locations. Where and how food production persists are central questions for the future of food security in an urbanising world. Our proposed framework provides directions for future research and highlights the role of policy and planning in reconciling food production with urban growth.


Racial inequalities and death on the horizon: COVID-19 and structural racism.
COVID-19 incidence and mortality in countries with heavy social inequalities differ in population terms. In countries like Brazil with colonial histories and traditions, the social markers of differences are heavily anchored in social and racial demarcation, and the political and social dynamics and processes based on structural racism act on this demarcation. The pandemic's actual profile in Brazil clashes with narratives according to which COVID-19 is a democratic pandemic, an argument aligned with the rhetoric of racial democracy that represents a powerful strategy aimed at maintaining the subaltern place of racialized populations such as indigenous peoples and blacks, as a product of modern coloniality. This essay focuses on the pandemic's profile in the Brazilian black population, in dialogue with decolonial contributions and critical readings of racism. The authors discuss government responses and COVID-19 indicators according to race/color, demonstrating the maintenance of historical storylines that continue to threaten black lives. The article also discusses the importance of local resistance movements, organized in the favelas, precarious urban spaces underserved by the State and occupied by black Brazilians.


Topiramate monotherapy in newly diagnosed epilepsy in children and adolescents.
A double-blind, dose-controlled study evaluated topiramate as monotherapy in 470 patients with newly diagnosed (< or = 3 months) epilepsy or epilepsy relapse in the absence of therapy. In addition to having at least 2 lifetime-unprovoked seizures, patients had 1 or 2 partial-onset seizures or generalized-onset tonic-clonic seizures during a 3-month retrospective baseline. The trial included a large cohort (N = 151, 32%) of children and adolescents 6 to 15 years of age. Eligible patients were randomized to treatment groups in which topiramate was titrated to target maintenance dosages of either 400 mg/day (n = 77) or 50 mg/day (n = 74). Patients were followed for at least 6 months. Based on Kaplan-Meier analyses, the primary efficacy endpoint of time to first seizure favored the higher topiramate dose in both the overall population and the cohort of children/adolescents. The probability that children/adolescents remaining in the study were seizure free at 6 months was 78% in the 50-mg target dose group and 90% with the higher dose. At 12 months, the probability of being seizure free was 62% and 85%, respectively. The incidence of treatment-limiting adverse events was 4% in the 50-mg target dose group and 14% in the group assigned to 400 mg as a target dose. The most common adverse events, excluding typical childhood illnesses, were headache, appetite decrease, weight loss, somnolence, dizziness, concentration/attention difficulty, and paresthesia. As shown in this subset analysis, topiramate is effective and well tolerated as monotherapy in children and adolescents.


Application of next-generation sequencing technology to diagnosis and treatment of focal segmental glomerulosclerosis.
A broad range of genetic and non-genetic factors can lead to kidney injury that manifests as focal segmental glomerulosclerosis (FSGS), which can be classified into primary (idiopathic) and secondary forms. Previous genetic approaches to familial or sporadic cases of FSGS or steroid-resistant nephrotic syndrome identified causal mutations in a subset of genes. Recently, next-generation sequencing (NGS) approaches are becoming a part of a standard assessment in medical genetics. Current knowledge of the comprehensive genomic information is changing the way we think about FSGS and draws attention not only to identification of novel causal genes, but also to potential roles for combinations of mutations in multiple genes, mutations with complex inheritance, and susceptibility genes with variable penetrance carrying relatively minor but significant effects. This review provides an update on recent advances in the genetic analysis of FSGS and highlights the potential as well as the new challenges of NGS for diagnosis and mechanism-based treatment of FSGS.


Experimental signatures of the mixed axial-gravitational anomaly in the Weyl semimetal NbP.
The conservation laws, such as those of charge, energy and momentum, have a central role in physics. In some special cases, classical conservation laws are broken at the quantum level by quantum fluctuations, in which case the theory is said to have quantum anomalies. One of the most prominent examples is the chiral anomaly, which involves massless chiral fermions. These particles have their spin, or internal angular momentum, aligned either parallel or antiparallel with their linear momentum, labelled as left and right chirality, respectively. In three spatial dimensions, the chiral anomaly is the breakdown (as a result of externally applied parallel electric and magnetic fields) of the classical conservation law that dictates that the number of massless fermions of each chirality are separately conserved. The current that measures the difference between left- and right-handed particles is called the axial current and is not conserved at the quantum level. In addition, an underlying curved space-time provides a distinct contribution to a chiral imbalance, an effect known as the mixed axial-gravitational anomaly, but this anomaly has yet to be confirmed experimentally. However, the presence of a mixed gauge-gravitational anomaly has recently been tied to thermoelectrical transport in a magnetic field, even in flat space-time, suggesting that such types of mixed anomaly could be experimentally probed in condensed matter systems known as Weyl semimetals. Here, using a temperature gradient, we observe experimentally a positive magneto-thermoelectric conductance in the Weyl semimetal niobium phosphide (NbP) for collinear temperature gradients and magnetic fields that vanishes in the ultra-quantum limit, when only a single Landau level is occupied. This observation is consistent with the presence of a mixed axial-gravitational anomaly, providing clear evidence for a theoretical concept that has so far eluded experimental detection.


Effect of Right Posterior Bile Duct Anatomy on Biliary Complications in Patients Undergoing Right Lobe Living Donor Liver Transplant.
Our aim was to evaluate the influence of the localization of right posterior bile duct anatomy relative to portal vein of the donors on posttransplant bile duct complications. We retrospectively investigated 141 patients who had undergone living donor liver transplant using right hemiliver grafts. The patients were classified based on the pattern of the right posterior bile duct and divided into infraportal and supraportal types. Clinical donor and recipient risk factors and surgical outcomes were compared for their relationship with biliary complications using logistic regression analyses. The 2 groups were similar according to demographic and clinical features. The biliary complication rate was 23.7% (9/38) in the infraportal group and 47.4% (37/78) in the supraportal group (P = .014). An analysis of risk factors for the development of anastomotic bile leak using logistic regression showed that a supraportal right posterior bile duct anatomy was a statistically significant positive predictor, with odds ratio of 18.905 (P = .012; confidence interval, 1.922-185.967). The distance of the right posterior bile duct from confluence was significantly lower in patients with biliary complications than in those without (mean of 7.66 vs 0.40 mm; P = .044). According to receiver operating characteristic analyses, the cut-off point for the length of right bile duct to right posterior bile duct from the hepatic confluence was 9.5 mm regarding presence of complications. Factors influencing bile duct anastomosis leakage were supraportal-type donor bile duct anatomy and length of the right main bile duct from biliary confluence. Hepatic arterial complications were similarly a risk factor for biliary strictures. Because of the multiple factors leading to complications in living donor liver transplant, it is challenging to group these patients by operative risk; however, establishing risk models may facilitate the prediction of complications.


Motivation and Prior Animal Experience of Newly Enrolled Veterinary Nursing Students at two Irish Third-Level Institutions.
Veterinary nurses report an intrinsic desire to work with animals. However, this motivation may be eroded by poor working conditions and low pay, resulting in the exit of experienced veterinary nurses from clinical practice. This study sought to quantify the level of animal-handling experience students possessed at the start of their training and to explore the factors motivating them to enter veterinary nurse training in two Irish third-level institutions. The authors had noted a tendency for veterinary nursing students to possess limited animal-handling skills, despite their obvious motivation to work with animals. The study explores possible reasons for this, as it mirrors previous reports in relation to students of veterinary medicine. First-year veterinary nursing students at Dundalk Institute of Technology and University College Dublin were surveyed and a focus group was held in each institution to explore student motivations for choosing this career and their prior animal-handling experience and workplace exposure. The results show that veterinary nursing students are highly intrinsically motivated to work with and care for animals. The majority had spent time in the veterinary workplace before starting their studies but they had limited animal-handling experience beyond that of family pets, primarily dogs. The study also revealed potential tensions between the veterinary nursing and veterinary medical students at University College Dublin: a hitherto unexposed aspect of the hidden curriculum in this institution. The results of this study highlight the need for ongoing investment in practical animal-handling training for veterinary nursing students.


Metabolism of the chemopreventive retinoid N-(4-hydroxyphenyl)retinamide by mammary gland in organ culture.
N-(4-Hydroxyphenyl)retinamide (4-HPR) is considered to be the most effective chemopreventive retinoid for chemically induced mammary carcinogenesis in rats. However, the mechanism of 4-HPR action in mammary cells is poorly understood. In the present study we examined the metabolism of 4-HPR in the mouse mammary gland in organ culture. Mammary glands excised from BALB/c mice were incubated with 4-HPR in the presence of insulin, prolactin and steroid hormones for 6 days. The glands were extracted with chloroform/methanol (2:1, v/v), and the metabolites were separated on a reversed-phase h.p.l.c. column. Three metabolites were separated in addition to 4-HPR; one of the metabolites, M2, was co-eluted with 13-cis-4-HPR, M3 was co-eluted with N-(4-methoxyphenyl)retinamide (4-MPR) and M1 remains unidentified. There appeared to be some hormonal regulation in the distribution of metabolites in the glands. Increased levels of 4-MPR and M1 were observed in insulin-plus-prolactin-treated glands as compared with the glands incubated with steroid hormones. Furthermore, it was observed that M1 isolated from the livers of 4-HPR-treated rats competed for the cellular retinoic acid-binding protein (CRABP) sites; however, 4-HPR did not bind to CRABP. These results indicate that mouse mammary gland can metabolize 4-HPR and that the metabolites which compete for CRABP sites may have physiological significance in the retinoid inhibition of mammary carcinogenesis.


An Analysis of 20 Cases of Radiation-Associated Sarcoma, Including 4 Cases Treated by Carbon Ion Radiotherapy.
Radiation-associated sarcoma (RAS) is one of the most life-threatening complications associated with the treatment of malignant neoplasms. Because all RAS patients have a history of radiotherapy, there have been no effective treatment options when RAS is not completely resected. We retrospectively reviewed 20 RAS patients, including 4 unresectable cases treated by carbon ion radiotherapy (CIRT). The primary diseases targeted by radiotherapy included malignant lymphoma (n = 4), cervical cancer (n = 3), pharyngeal cancer (n = 3), breast cancer (n = 2), lung cancer (n = 1), rectal cancer (n = 1), maxillary cancer (n = 1), synovial sarcoma (n = 1), and benign neoplasms (n = 4). The histological diagnoses of RAS included osteosarcoma (n = 8), leiomyosarcoma (n = 3), undifferentiated pleomorphic sarcoma (n = 3), rhabdomyosarcoma (n = 1), angiosarcoma (n = 1), malignant peripheral nerve sheath tumor (n = 1), spindle cell sarcoma NOS (n = 1), and sarcoma not further specified (n = 2). The median survival time from the diagnosis of RAS was 26 months. Eleven patients underwent surgery. Five of these patients achieved a continuous disease free (CDF) status or showed no evidence disease. Four patients underwent CIRT. One of these patients with leiomyosarcoma achieved a CDF status, and the other patient with osteosarcoma achieved a partial response. On the other hand, 2 patients experienced grade 3 toxicities that required surgical treatment. RAS originates from various types of diseases that are treated by radiotherapy and shows diverse pathological features. Complete resection achieves a good prognosis. CIRT can be an effective and feasible option for unresectable RAS.


Identification of D-peptide ligands through mirror-image phage display.
Genetically encoded libraries of peptides and oligonucleotides are well suited for the identification of ligands for many macromolecules. A major drawback of these techniques is that the resultant ligands are subject to degradation by naturally occurring enzymes. Here, a method is described that uses a biologically encoded library for the identification of D-peptide ligands, which should be resistant to proteolytic degradation. In this approach, a protein is synthesized in the D-amino acid configuration and used to select peptides from a phage display library expressing random L-amino acid peptides. For reasons of symmetry, the mirror images of these phage-displayed peptides interact with the target protein of the natural handedness. The value of this approach was demonstrated by the identification of a cyclic D-peptide that interacts with the Src homology 3 domain of c- SRC. Nuclear magnetic resonance studies indicate that the binding site for this D-peptide partially overlaps the site for the physiological ligands of this domain.


Expansion of targetable sites for the ribonucleoprotein-based CRISPR/Cas9 system in the silkworm Bombyx mori.
With the emergence of CRISPR/Cas9 technology, multiple gene editing procedures became available for the silkworm. Although binary transgene-based methods have been widely used to generate mutants, delivery of the CRISPR/Cas9 system via DNA-free ribonucleoproteins offers several advantages. However, the T7 promoter that is widely used in the ribonucleoprotein-based method for production of sgRNAs in vitro requires a 5' GG motif for efficient initiation. The resulting transcripts bear a 5' GG motif, which significantly constrains the number of targetable sites in the silkworm genome. In this study, we used the T7 promoter to add two supernumerary G residues to the 5' end of conventional (perfectly matched) 20-nucleotide sgRNA targeting sequences. We then asked if sgRNAs with this structure can generate mutations even if the genomic target does not contain corresponding GG residues. As expected, 5' GG mismatches depress the mutagenic activity of sgRNAs, and a single 5' G mismatch has a relatively minor effect. However, tests involving six sgRNAs targeting two genes show that the mismatches do not eliminate mutagenesis in vivo, and the efficiencies remain at useable levels. One sgRNA with a 5' GG mismatch at its target performed mutagenesis more efficiently than a conventional sgRNA with 5' matched GG residues at a second target within the same gene. Mutations generated by sgRNAs with 5' GG mismatches are also heritable. We successfully obtained null mutants with detectable phenotypes from sib-mated mosaics after one generation. In summary, our method improves the utility and flexibility of the ribonucleoprotein-based CRISPR/Cas9 system in silkworm.


Hydroxychloroquine and Remdesivir in COVID-19: A critical analysis of recent events.
The world is going through an unprecedented medical emergency with no effective remedy for the SARS-CoV2 virus causing Covid-19. Two drugs used for other indications in the past, hydroxychloroquine (HCQ) and remdesivir (RDV), are sought to be repurposed to treat Covid-19. Both these drugs have received emergency use authorisation by the US Food and Drug Administration. In this review, we critically analyse the identification of and subsequent events concerning these two drugs as potential treatment options for Covid-19, and conclude by raising some ethical issues that require serious thought from the global scientific community concerned with using these two drugs against Covid-19.

 Key Words: Covid-19, hydroxychloroquine, remdesivir, USFDA, emergency use authorisation

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The value of the study of natural history in genetic disorders and congenital anomaly syndromes.
The study of the natural history of genetic disorders and syndromes with congenital anomalies and dysmorphic features is a challenging and often neglected area. There are many reasons to pursue this type of research but it requires special clinical skills and a considerable amount of hard work. Setting up protocols and collecting data is complex and time consuming. Frequently, helpful clues for a particular disorder come from the study of the natural history of other disorders. Older affected subjects and unique cases with unusual features are often most important in unravelling the 'normal' course of a disease or recognising the basic defect. The study of natural history from individual patients and their records is complementary to population or registry based studies because it identifies individual variations and clinical heterogeneity. The understanding of the natural history of a particular disorder is of importance both to the affected person and their family and to the physicians caring for them. It is also useful to the basic researcher trying to determine the pathogenetic mechanism causing the disorder. In many ways, clinical geneticists have learned the art of caring for patients, as well as the challenges of clinical genetics, by becoming apprentices to and studying in depth specific disease entities.


Impact of interventions designed to reduce medication administration errors in hospitals: a systematic review.
There is a need to identify effective interventions to minimize the threat posed by medication administration errors (MAEs). Our objective was to review and critically appraise interventions designed to reduce MAEs in the hospital setting. Ten electronic databases were searched between 1985 and November 2013. Randomized controlled trials (RCTs) and controlled trials (CTs) reporting rates of MAEs or related adverse drug events between an intervention group and a comparator group were included. Data from each study were independently extracted and assessed for potential risk of bias by two authors. Risk ratios (RRs, with 95 % confidence intervals [CIs]) were used to examine the effect of an intervention. Six RCTs and seven CTs were included. Types of interventions clustered around four main themes: medication use technology (n = 4); nurse education and training (n = 3); changing practice in anesthesia (n = 2); and ward system changes (n = 4). Reductions in MAE rates were reported by five studies; these included automated drug dispensing (RR 0.72, 95 % CI 0.53-1.00), computerized physician order entry (RR 0.51, 95 % 0.40-0.66), barcode-assisted medication administration with electronic administration records (RR 0.71, 95 % CI 0.53-0.95), nursing education/training using simulation (RR 0.17, 95 % CI 0.08-0.38), and clinical pharmacist-led training (RR 0.76, 95 % CI 0.67-0.87). Increased or equivocal outcome rates were found for the remaining studies. Weaknesses in the internal or external validity were apparent for most included studies. Theses and conference proceedings were excluded and data produced outside commercial publishing were not searched. There is emerging evidence of the impact of specific interventions to reduce MAEs in hospitals, which warrant further investigation using rigorous and standardized study designs. Theory-driven efforts to understand the underlying causes of MAEs may lead to more effective interventions in the future.


Polarized light microscopy in reproductive and developmental biology.
The polarized light microscope reveals orientational order in native molecular structures inside living cells, tissues, and whole organisms. It is a powerful tool used to monitor and analyze the early developmental stages of organisms that lend themselves to microscopic observations. In this article, we briefly discuss the components specific to a traditional polarizing microscope and some historically important observations on: chromosome packing in the sperm head, the first zygote division of the sea urchin, and differentiation initiated by the first asymmetric cell division in the sand dollar. We then introduce the LC-PolScope and describe its use for measuring birefringence and polarized fluorescence in living cells and tissues. Applications range from the enucleation of mouse oocytes to analyzing the polarized fluorescence of the water strider acrosome. We end with new results on the birefringence of the developing chick brain, which we analyzed between developmental stages of days 12-20.


Culturable fungi of stored 'golden delicious' apple fruits: a one-season comparison study of organic and integrated production systems in Switzerland.
The effects of organic and integrated production systems on the culturable fungal microflora of stored apple fruits from five matched pairs of certified organic and integrated 'Golden Delicious' farms were studied at five representative production sites in Switzerland. Isolated fungi were identified morphologically. Colonization frequency (percentage of apples colonized), abundance (colony numbers), and diversity (taxon richness) were assessed for each orchard. The standard quality of the stored fruits was comparable for both organic and integrated apples and complied with national food hygiene standards. Yeasts (six taxa) and the yeast-like fungus Aureobasidium pullulans were the dominant epiphytes, filamentous fungi (21 taxa) the dominant endophytes. The most common fungi occurred at all sites and belonged to the "white" and "pink" yeasts, yeast-like A. pullulans, filamentous fungi Cladosporium spp., Alternaria spp., and sterile filamentous fungi. Canonical correspondence analysis of the total fungal community revealed a clear differentiation among production systems and sites. Compared to integrated apples, organic apples had significantly higher frequencies of filamentous fungi, abundance of total fungi, and taxon diversity. The effects of the production system on the fungal microflora are most likely due to the different plant protection strategies. The incidence of potential mycotoxin producers such as Penicillium and Alternaria species was not different between production systems. We suggest that higher fungal diversity may generally be associated with organic production and may increase the level of beneficial and antagonistically acting species known for their potential to suppress apple pathogens, which may be an advantage to organic apples, e.g., in respect to natural disease control.


Efficacy of a regional poisoning treatment centre in preventing further suicidal behaviour.
A survey of parasuicide (attempted suicide) in general practice in Edinburgh allowed comparison of the rates of further suicidal behaviour in patients treated for their initial episode in the Edinburgh Regional Poisoning Treatment Centre and in those referred to psychiatrists elsewhere or not referred at all. Further suicidal behaviour was only one-third as common among those treated in the treatment centre as among non-admitted patients; patients referred to other psychiatric services did no better than those who were not referred to a psychiatrist at all. The difference in repetition frequencies could not be accounted for in terms of selection of patients in the treatment centre who were less likely to repeat. The tentative conclusion is made that crisis intervention of the kind available at the Edinburgh centre is effective in secondary prevention. The findings add support to Government recommendations that special units like the one in Edinburgh should be set up in other regions.


The molecular origins of evolutionary innovations.
The history of life is a history of evolutionary innovations, qualitatively new phenotypic traits that endow their bearers with new, often game-changing abilities. We know many individual examples of innovations and their natural history, but we know little about the fundamental principles of phenotypic variability that permit new phenotypes to arise. Most phenotypic innovations result from changes in three classes of systems: metabolic networks, regulatory circuits, and macromolecules. I here highlight two important features that these classes of systems share. The first is the ubiquity of vast genotype networks - connected sets of genotypes with the same phenotype. The second is the great phenotypic diversity of small neighborhoods around different genotypes in genotype space. I here explain that both features are essential for the phenotypic variability that can bring forth qualitatively new phenotypes. Both features emerge from a common cause, the robustness of phenotypes to perturbations, whose origins are linked to life in changing environments.


Term infant studies of DHA and ARA supplementation on neurodevelopment: results of randomized controlled trials.
Healthy term infants who are not breast-fed may need long-chain polyunsaturated fatty acids (LCPUFA) in their feeding, based on the changes in plasma and tissue fatty composition. However, consistent functional effects across different studies conducted over the past two decades has been more difficult to document. The interpretation of these data has scientific and public interest with the introduction of LCPUFA supplemented formula. There are 14 controlled trials in term infants that have included formula feeding with or without LCPUFA and functional assessment of visual and other measures of neural development; in addition, 7 have evaluated specific measures related to cognitive development. We chose to examine the effect of DHA dose provided daily on the development of visual acuity to explain the differences in visual acuity responses across randomized studies. A "meta-regression" was performed with the use of a DHA effective dose as the independent variable and visual acuity at 4 months as the dependent variable. Since the two main dietary determinants of DHA status are the LNA provided and the preformed DHA consumed, we defined DHA equivalent dose across studies by assuming a 1%, 5%, and 10% conversion of LNA to DHA. Results indicate a strong and significant effect of DHA equivalent dose on magnitude of the visual acuity response at all conversions tested; greatest significance was found when using a 10% bioequivalency (r(2)=0.68, and P=.001). We conclude that there is a significant relation between the total DHA equivalents provided and effectiveness as defined by visual acuity measurements at 4 months of age.


Evaluation of antibody persistence after a four-dose primary hepatitis B vaccination and anamnestic immune response in children under 6 years.
Introduction. Tunisia is an intermediate hepatitis B virus (HBV) endemic country. The vaccination against hepatitis B was introduced in 1995 including four doses with a first dose administrated at birth. Decreasing the level of antibodies against hepatitis B surface antigen (anti-HBs) over time can be alarming. This study was conducted to explore the anti-HBV immune response among children under 6 years old, vaccinated according to the national vaccination schedule, by evaluating the immunological response to primary vaccination and by exploring the anamnestic immune response to a booster dose.Methods. We conducted a cross-sectional prospective study from June 2016 to June 2017 (n=180), based on voluntary participation. Children were recruited from the public pediatric ward sectors in Sahloul University Hospital of Sousse in Central Tunisia. An anti-HB titre was determined based on electro-chemiluminescence micro-particle immunoassay (ECLIA), using Elecsys Anti-HBs II kit, Roche.Results. Mean age at the time of enrollment in the study was 33±14.8 months. The seroprotection rate was 77.2 %. The anti-HB titre differed significantly between the different age groups (P=0.002). The predicting variable for having no seroprotective antibody level was older age. Children with anti-HB levels <10 IU l- 1 were offered an additional dose of HBV vaccine. Anamnestic response 1 month after the challenge dose was observed in 100 % of subjects. The probability of developing a high antibody response, following the booster dose increased in conjunction with an increased pre-booster antibody level.Conclusion. The response to a booster dose suggests the persistence of immune memory in almost all vaccinated individuals. Although a booster dose increases substantially anti-HB titre, the clinical relevance of such an increase remains unknown.


Quality of life and cost factors in panic disorder.
Quality of life encompasses domains of personal happiness, role fulfillment, and health status. Increasing attention has been paid to the relationship between quality of life and panic disorder, with accumulating evidence now available to suggest impairment in several domains among subjects with panic disorder. This review summarizes the results of community-based and treatment-seeking populations of subjects with panic disorder. Impaired personal happiness, restricted role functioning, and increased use of health services are all described. Evidence suggests that accurate diagnosis and effective treatment can significantly lessen health service utilization, resulting in substantial cost offset and also leading to increased work productivity and personal effectiveness.


A new genus of bamboo culm tarantula from Thailand (Araneae, Mygalomorphae, Theraphosidae).
Bamboo plays an important role in the animal world, including providing a nutritious food source, shelter, and habitat. Inside of bamboo culm, we discovered a new genus of tarantula, which we describe here as Taksinus gen. nov. (♂♀). Specimens of this new tarantula were collected from Mae Tho, Mueang Tak district, Tak province, in Thailand, making it geographically distant from any other arboreal genera. Genital morphology was used to diagnose its genus, which is supported by distributional data, natural history, morphological characters, and photographic illustrations of the male and female. Diagnosis of the new genus was determined by distinguishing its different characters from those of other arboreal theraphosid spiders distributed throughout Southeast Asia. This tarantula's specialization is that it lives in the stalks of the Asian bamboo Gigantochloa sp.


Evaluation of non-adherence to anti-retroviral therapy, the associated factors and infant outcomes among HIV-positive pregnant women: a prospective cohort study in Lesotho.
Success in addressing prevention of mother-to-child transmission of HIV depends largely on good adherence to anti-retroviral therapy (ART) by pregnant women. Knowledge of the levels of ART adherence among pregnant women is essential to inform strategies to prevent or reduce HIV transmission rates, particularly in African settings. Aim: the primary objective of this study was to measure adherence to anti-retroviral therapy (ART) among pregnant women living with human immunodeficiency virus (HIV). The secondary objectives were to determine: i) the rate of new infections among children at Mabote Filter Clinic in Maseru, Lesotho whose mothers were enrolled in PMTCT, and ii) the factors associated with non-adherence to ART among pregnant women. In this prospective cohort study, HIV-positive pregnant women receiving antiretroviral therapy (ART) for prevention of mother to child transmission (PMTCT) were followed up to delivery and their children were tested for HIV. We collected socio-demographic information, knowledge of PMTCT and adherence to ART (three-day recall and pill count) including reasons for non-adherence. We also used logistic regression to explore factors associated with non-adherence. One hundred and seven women were included. The mean (standard deviation) age of the participants was 28.2 (5.7) years. Most, 81.3% (87/107), were married, only 9.3% (10/107) had a postsecondary education. Two-thirds (63.6%: 68/107) of the participants started ART because of PMTCT. Only 78.5% (84/107) of the participants had adequate knowledge of the importance of PMTCT. The three-day self-reported non-adherence rate at the first visit was 7.5% (95% confidence interval (CI): 3.7, 13.1), but up to 43.4% (95% CI: 35.2, 51.9) using pill count. The most frequently reported reasons for not adhering were: running out of pills (7.5%), nausea (5.6%) and to avoid side-effects (3.7%). Women who were employed (odds ratio (OR) 4.35; 95% CI: 1.38,14.29; p = 0.012) and at a higher gestational age (OR = 1.43; 95% CI: 1.11, 1.85; p = 0.006) were more likely to be non-adherent. Only 1 of the 77 exposed infants was found to be positive for HIV at 6 weeks after birth. We found a higher non-adherence rate for participants with pill count compared to a three-day adherence self-report. However, mother to child HIV transmission was relatively low. Lack of employment and relatively high gestational age were found to be predictive factors of non-adherence.


Electrotactile feedback outweighs natural feedback in sensory integration during control of grasp force.
Objective.The nervous system subconsciously estimates the state of the body as a weighted average of the information from various sensory sources, where the weights reflect the perceived reliability of each source. Loss of motor functions can be partially compensated using assistive systems (e.g. prostheses), which may also restore somatosensory feedback through tactile stimulation. Whether such artificial feedback is integrated in the neural state estimation process is not known.Approach.In this study, able-bodied subjects performed a grasp force matching task with supplementary non-invasive electrotactile stimulation with a frequency proportional to grasp force magnitude. Before the task, a brief training session taught the subjects to associate the sensation of electrotactile stimulation with the generated grasp force. In some trials, the force-frequency mapping was biased to introduce an unnoticeable mismatch between natural and electrotactile force feedback, thereby provoking the subject to subconsciously estimate the force as a compromise between the two sources of information.Main results.The outcome of this compromise revealed the weights assigned to each feedback type. The grasp forces were significantly affected by the biased mappings, as indicated by the average estimated relative weights (electrotactile: 0.69 ± 0.29; natural: 0.31 ± 0.29). Across subjects, this weight was correlatedr2=0.75) with the improvement in force matching precision when adding the unbiased electrotactile feedback to the natural force feedback, as predicted by maximum likelihood estimation. This shows that even after minimal training the nervous system adopts electrotactile stimulation as a highly reliable source of information that can improve the precision in the estimation of the grip force.Significance.This result has important implications for the restoration of sensory feedback in upper limb prostheses as it indicates that even non-invasive stimulation can be integrated naturally (i.e. subconsciously and effectively) in the motor controlloop.


Adult heart disease in Mt Hagen: a study of 154 patients.
Although several previous studies have been done on the nature and prevalence of cardiovascular disease in Papua New Guinea no study has looked exclusively at a highlander population. This article reviews 154 cardiac patients who first presented to the Mt Hagen adult outpatient department over a period of one year. The study excluded non-highlanders, patients under 12 years of age, and patients with heart disease secondary to anaemia or diseases of the blood vessels. Heart disease was found to constitute a significant proportion of outpatient visits and admissions. Cor pulmonale secondary to chronic lung disease was the commonest condition seen, occurring in higher frequency than reported elsewhere, and accounting for the majority of cases of congestive heart failure. Valvular heart disease was also common, often presenting in a precocious and severe form. Congenital bicuspid aortic values were important in the generation of aortic valve disease in this population. Arrhythmias and conduction disturbances were also common. Diseases of the myocardium and pericardium occurred infrequently and were of the same nature as those reported in other studies in Papua New Guinea. Hypertension was probably underreported in this study, with renal disease being a contributing factor in the cases seen. Ischaemic heart disease represented a small number of the total cases, but was probably underreported.


Scottish Confidential Inquiry into Asthma Deaths (SCIAD), 1994-6.
There have been important changes in the organisation of care for patients with asthma since asthma deaths were studied in the 1980s by the British Thoracic Association (BTA), with greater emphasis on long term control of symptoms and the use of preventive therapy. Recent trends in routine statistics show a decline in population death rates. A confidential review was undertaken of general practice and hospital records and interviews with general practitioners of patients dying in mainland Scotland between January 1994 and December 1996 with a principal diagnosis of asthma recorded by the Registrar General's Office. Panel assessment of the cause of death was carried out and a number of possible adverse factors were identified. The data from the 15-64 year age group were compared with similar data from the earlier study by the BTA. Over the three year period 95 deaths of 235 studied (40%) were confirmed as being due to asthma. Taking account of different methods of case ascertainment used in the BTA and this study, a fall in the calculated rate of "deaths assessed as due to asthma" was found from 2.51 (95% CI 2.34 to 2.68) per 100,000 population in 1979 to 1.26 (95% CI 1.19 to 1.33) per 100,000 population in 1994-6. Fewer individual adverse factors were identified in clinical management, with appropriate routine management in 59% and management of the final attack satisfactory in 71%. Patient factors such as poor compliance, lack of peak expiratory flow (PEF) measurements, and overuse of reliever medication without inhaled corticosteroids, and psychosocial problems, notably depression, were confirmed as important contributing factors. Four of five patients under 16 years of age who died were found to have problems with routine management. This population based study documents important improvements in the standard of asthma care as well as a significant decline in the rate of deaths due to asthma over a period during which the organisation of care has changed and the chronic nature of the disease has been acknowledged. Strategies which might have a further impact include the greater use of PEF recordings, particularly during acute attacks, to document recovery, prescription monitoring of the underuse of inhaled corticosteroids, consideration of the use of combined preparations where persistent overuse of bronchodilators is occurring, and increased input for young patients whose routine management is proving difficult.


Postoperative contrast enhancement in patients with brain tumor.
Contrast enhancement resulting from surgical trauma may mimic residual enhancing tumor, thereby complicating the interpretation of postoperative computed tomographic scans. We assessed the natural history of postoperative enhancement in 10 patients with brain tumor. Contrast enhancement distinguishable from residual enhancing tumor appeared along the operative margin following tumor resections but not lobectomies. Enhancement appeared as early as the fifth postoperative day, was most intense at two weeks, and persisted for several months. Prior to the fifth postoperative day, enhancement reflected residual tumor. Edema and artifacts were more prominent on the first and second postoperative days than on the third and fourth. We recommend that postoperative computed tomographic scans to assess residual enhancing tumor be performed on the third or fourth postoperative day. This timing avoids postoperative enhancement and minimizes interpretative difficulties caused by artifacts.


Once versus thrice daily tobramycin alone and in combination with ceftazidime, ciprofloxacin and imipenem in an in vitro pharmacodynamic model.
The purpose of this study was to compare once daily (To24) and thrice daily (To8) tobramycin dosing regimens alone and in combination with ceftazidime, ciprofloxacin and imipenem against a clinical and ATCC strain of Pseudomonas aeruginosa. A one-compartment in vitro pharmacodynamic model was used to simulate bacteremic infection. Pharmacodynamic parameters, including rate of bacterial kill following the first dose or time to achieve a 99.9% reduction in bacterial counts (T99.9%), extent of bacterial kill with subsequent doses and bacterial count at 24 h were characterized for each regimen. Compared to To8 alone, the combination regimens had a shorter T99.9% (p = 0.045) and greater extent of bacterial kill following the first dose (p = 0.045). Compared to To24 alone, the combination regimens demonstrated similar rates (p = 0.067) and extents of bacterial kill following the first dose (p = 0.32), however, produced lower bacterial counts at 24 h (p = 0.045). The various combination regimens appeared equally effective considering rate and extent of bacterial kill following the first dose and residual inocula at 24 h. Ciprofloxacin-containing regimens demonstrated the most bacterial kill with subsequent doses, however, bacterial counts at 24 h were similar to those of other combination regimens.


[The use of computers and mathematical methods for optimizing epidemic control measures with respect to measles infection].
The potentialities of computers for the study of the effectiveness of immunization have been demonstrated and the mathematical model for the prediction of the proportion of children, seronegative to measles, derived on the basis of the data on the average measles morbidity in different groups for a given period. A multifactor analysis of a large scope of data obtained in seroepidemiological survey and a retrospective analysis of measles morbidity on the basis of data collected in two districts of Moscow have been carried out with the use of computers and mathematical methods.


An essential role of the basal body protein SAS-6 in Plasmodium male gamete development and malaria transmission.
Gametocytes are the sole Plasmodium parasite stages that infect mosquitoes; therefore development of functional gametes is required for malaria transmission. Flagellum assembly of the Plasmodium male gamete differs from that of most other eukaryotes in that it is intracytoplasmic but retains a key conserved feature: axonemes assemble from basal bodies. The centriole/basal body protein SAS-6 normally regulates assembly and duplication of these organelles and its depletion causes severe flagellar/ciliary abnormalities in a diverse array of eukaryotes. Since basal body and flagellum assembly are intimately coupled to male gamete development in Plasmodium, we hypothesized that SAS-6 disruption may cause gametogenesis defects and perturb transmission. We show that Plasmodium berghei sas6 knockouts display severely abnormal male gametogenesis presenting reduced basal body numbers, axonemal assembly defects and abnormal nuclear allocation. The defects in gametogenesis reduce fertilization and render Pbsas6 knockouts less infectious to mosquitoes. Additionally, we show that lack of Pbsas6 blocks transmission from mosquito to vertebrate host, revealing an additional yet undefined role in ookinete to sporulating oocysts transition. These findings underscore the vulnerability of the basal body/SAS-6 to malaria transmission blocking interventions.


Physiological and morphological characterization of dentate granule cells in the p35 knock-out mouse hippocampus: evidence for an epileptic circuit.
There is a high correlation between pediatric epilepsies and neuronal migration disorders. What remains unclear is whether there are intrinsic features of the individual dysplastic cells that give rise to heightened seizure susceptibility, or whether these dysplastic cells contribute to seizure activity by establishing abnormal circuits that alter the balance of inhibition and excitation. Mice lacking a functional p35 gene provide an ideal model in which to address these questions, because these knock-out animals not only exhibit aberrant neuronal migration but also demonstrate spontaneous seizures. Extracellular field recordings from hippocampal slices, characterizing the input-output relationship in the dentate, revealed little difference between wild-type and knock-out mice under both normal and elevated extracellular potassium conditions. However, in the presence of the GABA(A) antagonist bicuculline, p35 knock-out slices, but not wild-type slices, exhibited prolonged depolarizations in response to stimulation of the perforant path. There were no significant differences in the intrinsic properties of dentate granule cells (i.e., input resistance, time constant, action potential generation) from wild-type versus knock-out mice. However, antidromic activation (mossy fiber stimulation) evoked an excitatory synaptic response in over 65% of granule cells from p35 knock-out slices that was never observed in wild-type slices. Ultrastructural analyses identified morphological substrates for this aberrant excitation: recurrent axon collaterals, abnormal basal dendrites, and mossy fiber terminals forming synapses onto the spines of neighboring granule cells. These studies suggest that granule cells in p35 knock-out mice contribute to seizure activity by forming an abnormal excitatory feedback circuit.


5-Aza-2'-deoxycytidine increases sialyl Lewis X on MUC1 by stimulating β-galactoside:α2,3-sialyltransferase 6 gene.
Sialyl Lewis X is a tumor-associated antigen frequently found in the advanced cancers. However, the mechanism for the production of this cancer antigen is not entirely clear. The objective of this study is to examine whether epigenetics is involved in the regulation of the formation of this antigen. We observed an increase of sialyl Lewis X in HCT15 cells, a colon cancer cell line, treated with 5-Aza-2'-deoxycytidine. This treatment enhanced the expression of β-galactoside:α2,3-sialyltransferase 6 gene and sialyl Lewis X on MUC1, and the adherence of these cells to E-selectin under dynamic flow conditions. In addition, 5-Aza-2'-deoxycytidine treatment inhibited methylation of β-galactoside:α2,3-sialyltransferase 6 gene and siRNA knockdown of this gene drastically reduced sialyl Lewis X without affecting MUC1 expression. We conclude that 5-Aza-2'-deoxycytidine treatment increases sialyl Lewis X on MUC1 by stimulating the β-galactoside:α2,3-sialyltransferase 6 gene via inhibition of DNA methylation. Increased sialyl Lewis X by 5-Aza-2'-deoxycytidine raises a concern about the safety of this chemotherapeutic drug. In addition, β-galactoside:α2,3-sialyltransferase 6 gene may be a potential therapeutic target for suppressing tumorigenicity of colon cancer.