A dynamic,multi‐level analysis of recent immunization trends in Colombia

Abstract

During 1985–91, Latin American ministries of health carried out the ultimately successful Regional Polio Eradication Initiative. Unprecedented vaccine coverage levels were attained through a combination of mass campaigns, house‐to‐house vaccinations, and improvements in routine immunization services. Little is known, however, about the effects of these interventions on immunization demand; whether they reached so‐called high‐risk households and, if so, whether program effects were sufficient to offset the household risk factors. This paper examines the probability and timing of full immunization over this period in one case country, Colombia. Information on the immunization status of 3,609 vaccine‐eligible children born 1985–90 was extracted from Colombia's 1990 Demographic and Health Survey. Annual immunization coverage estimates from the Colombian Ministry of Health for 1985–90 for 148 sample municipios were appended to each child record, along with household‐level data. Initial non‐parametric regressions showed that five of six observed risk factors negatively influenced full immunization probability. Multivariate logit models showed that parents who had already lost a child were significantly less likely to obtain immunization cards (a proxy for exposure to the routine immunization program), despite rising cardholdership rates over the period. Among 1,376 immunization cardholders, waiting times to full immunization fell monotonically over the period. Local program coverage of 80 per cent or higher and prior use of prenatal services both increased the probability of full immunization. However, three of five maternal occupational categories decreased the probability, as did three of six observed household risk factors. The results show that demand for routine immunizations rose over the period, that only the highest‐risk households were not exposed to the routine program, and that routine program participation partially offset negative risk factor effects on the probability of full immunization. While targeted PHC interventions may increase health production by recruiting high‐risk households into the routine PHC services, further health production increases will require more intensive follow‐up of such households through routine PHC services.     

Temporal characteristics of transmission-line loadings in the Swedish childhood cancer study

A recent study conducted in Sweden reported that 1) leukemia risk in children who lived near 220 or 400 kV electric-power transmission lines was associated with calculated historical magnetic field levels; 2) children living within a distance of 50 m of transmission lines had an elevated risk of leukemia; and 3) there was no association between leukemia and residential magnetic fields measured many years after diagnosis. Subsequently, these investigators found through logistic regression analysis that disease was more strongly associated with calculated historical fields than with distance. Since the calculated historical fields in that study depended predominantly on distance and transmission-line load current, the logistic regression results suggest that historical load current plays an important role in the epidemiological results. Thus, we studied hourly 1974 load-current data for six transmission lines, and we examined 1958–1985 annual load-current data for 112 transmission lines. Most lines exhibited marked diurnal load-current rhythms during 1974, and all six showed systematic weekday weekend differences. During 1958–1985, average loadings of Swedish 220 and 400 kV lines increased by about 1.3% year. Predictive-value and kappa-statistic analyses indicated that Swedish transmission-line load currents were not stable over long periods, so that contemporaneous load current (or a contemporary magnetic field measurement) was not a good surrogate for historical load current (or historical magnetic fields). The results provide a potential explanation of the failure of the Swedish Study to find an association between leukemia and contemporaneous magnetic field levels measured many years after the etiologic period, and suggest that the inclusion of load-current data could significantly improve the quality of historical field calculations. Bioelectromagnetics 19:354–365, 1998. © 1998 Wiley-Liss, Inc.     

Dating and rating of Harris's lines

Well-preserved skeletons of 50 Dutch whalers from the 17th and 18th centuries, buried on the island of Zeeusche Uytkyck , were excavated during the Spitsbergen Expedition 1980. In order to estimate the biological age of these skeletons at the time that Harris's lines were formed, a method which was developed from routine X-ray data on tibial growth was tested. Formation ages of diaphyseal lines and strong metaphyseal lines appear to correspond well with hypoplastic enamel deformations within the same individuals. Because of the involvement of both the bone and tooth developmental systems, and of the resistance of these lines to bone remodeling, it can be assumed that they generally are the result of serious health insults.     

High-throughput karyotyping of human pluripotent stem cells

Genomic integrity of human pluripotent stem cell (hPSC) lines requires routine monitoring. We report here that novel karyotyping assay, utilizing bead-bound bacterial artificial chromosome probes, provides a fast and easy tool for detection of chromosomal abnormalities in hPSC lines. The analysis can be performed from low amounts of DNA isolated from whole cell pools with simple data analysis interface. The method enables routine screening of stem cell lines in a cost-efficient high-throughput manner.     

Alterations in microtubule assembly caused by the microtubule-active drug LY195448

LY195448 is an experimental drug that blocks cells at metaphase (Boder et al.: Microtubules and Microtubule Inhibitors 1985: 353-361, 1985). A 4 hour exposure of NRK cells to a drug concentration of 46 microM (15 micrograms/ml) increased the number of mitotic cells in the population from 4.9% to 18.5%. Examination of treated cells by immunofluorescence showed increased numbers of cells blocked at prometaphase, with short microtubules extending from the spindle pole to the kinetochores. The cytoskeleton of interphase cells remained intact at these concentrations. However, the number of microtubules appeared to be reduced, and those that remained appeared kinkier and curled, particularly toward the periphery of the cells. When cytoskeletal microtubules of NRK cells were depolymerized with nocodazole, they reassembled within minutes of transfer to drug-free media. However, nocodazole-treated cells transferred to fresh media containing 15 micrograms/ml of LY195448 required 2-3 times longer to reassemble cytoplasmic microtubules. Previously isolated Chinese hamster ovary cell microtubule mutants resistant to either taxol or Colcemid were tested for cross-resistance to this drug. Cell lines resistant to the depolymerizing drug Colcemid exhibited increased resistance to LY195448 compared to wild-type cells, whereas taxol resistant cell lines were more sensitive. Of eleven newly isolated mutant CHO cell lines selected for increased resistance to LY195448, seven exhibited an altered beta-tubulin protein by two-dimensional polyacrylamide gel electrophoresis. These 11 cell lines also showed a heterogenous pattern of resistance to several microtubule-active drugs. These data demonstrate that LY195448 is cytotoxic to mammalian cells because it inhibits microtubule assembly, most likely through a direct interaction with tubulin.     

Culture-based preservation of Marchantia polymorpha gemmalings and thalli without encapsulation,drying, or freezing

Ongoing research has generated many important lines of the model liverwort Marchantia polymorpha, including mutants and transgenic lines. To maintain these lines, researchers typically spend a lot of time and effort periodically replanting thalli (e.g., every month). To avoid this routine maintenance, researchers have developed methods for cryopreservation of dried and frozen gemmae. In this study, we developed a culture-based method for preserving gemmalings and thalli without encapsulation, drying, or freezing. The method requires only tissue culture on agar medium supplemented with sucrose in the dark at regular temperature (22°C). These culture conditions severely inhibit growth of gemmalings and thalli; however, these tissues remained alive after more than 1 year of storage. Survival rate of tissues using this method was 100% in all tests. This method thus enables preservation of gemmaling and thallus cultures on medium under regular temperature conditions, thereby relieving researchers of labor-intensive routine maintenance.     

长白山自然保护区猫科动物种群数量变化及现状

1985～2009年,在长白山自然保护区及其周边地区采用野外样线调查和社会访问调查的方法,对猫科动物种群数量变化及影响因素进行了研究.结果表明,长白山自然保护区记载的东北虎(Pantheratigris)、远东豹(P.pardus)、猞猁(Felis lynx)和豹猫(F.bengalensis)4种猫科动物中,野外调...     

Quality control genotyping for assessment of genetic identity and purity in diverse tropical maize inbred lines

Quality control (QC) genotyping is an important component in breeding, but to our knowledge there are not well established protocols for its implementation in practical breeding programs. The objectives of our study were to (a) ascertain genetic identity among 2–4 seed sources of the same inbred line, (b) evaluate the extent of genetic homogeneity within inbred lines, and (c) identify a subset of highly informative single-nucleotide polymorphism (SNP) markers for routine and low-cost QC genotyping and suggest guidelines for data interpretation. We used a total of 28 maize inbred lines to study genetic identity among different seed sources by genotyping them with 532 and 1,065 SNPs using the KASPar and GoldenGate platforms, respectively. An additional set of 544 inbred lines was used for studying genetic homogeneity. The proportion of alleles that differed between seed sources of the same inbred line varied from 0.1 to 42.3?%. Seed sources exhibiting high levels of genetic distance are mis-labeled, while those with lower levels of difference are contaminated or still segregating. Genetic homogeneity varied from 68.7 to 100?% with 71.3?% of the inbred lines considered to be homogenous. Based on the data sets obtained for a wide range of sample sizes and diverse genetic backgrounds, we recommended a subset of 50–100 SNPs for routine and low-cost QC genotyping, verified them in a different set of double haploid and inbred lines, and outlined a protocol that could be used to minimize errors in genetic analyses and breeding.     

Correlation analysis between some blood properties and atmospheric environmental parameters

S.W. Tromp made investigations of a weather effect on erythrocyte sedimentation rates (ESR) of human blood by routine checks of the blood of donor groups in Leiden from 1955 to 1985. A higher ESR was found for the summer season and lower ESR for winter and low values occurred soon after a strong cooling spell. In this report, we have continued his work using data for the years 1971–1985 from Leiden (The Netherlands). An influence of the weather on ESR was also found, but this seems to be more complicated than Tromp supposed. Some new aspects are described and elaborated, e.g. ESR already increases before warming is noticed at the ground layer. This is possibly caused by changes in the upper atmosphere, a suggestion that requires further studies. For periods of low ESR, there was a greater number of occlusion fronts passing the Netherlands. The long-term fluctuations of ESR that were found were correlated with sun spot relative numbers only in a few periods. The results of our study justify further research for a variety of other locations around the world.     

VTUTIN: A full screen gel management editor

Large DNA sequences are now routinely sequenced by the cloningof randomly generated fragments into single-stranded DNA phagevectors (the ‘shotgun’ method). Various programsexist for computerized assembly of such fragments, includingthe phases of data entry, homology searching and gel-management/editing.Many gel-management editors are rudimentary in nature, usingeither line-editing techniques or using unnatural displays orcommand systems. Others are available only on restricted typesof computer system. The program VTUTIN makes full screen editingalong the lines of modern text editors available for the complexdata type of sets of sequence gels and their consensus. Notonly are the data displayed on the VDU screen in a natural manner,but VTUTIN has also been written to model the command systemof a well-established text editor (PDP-ll KED or VAX/VMS EDT)to simplify editor use and learning. VTUTIN has been writtenin Pascal in a modular form so that wide-spread portabilityis facilitated. VTUTIN is currently implemented to work on VT-100type terminals although the modularity of the code should allowstraightforward conversion for other terminal types and shouldalso permit simple alteration to model any other text editor. Received on July 5, 1985; accepted on September 23, 1985     

A proteomics approach to identifying fish cell lines

Fish cell lines are relatively easy to culture and most have simple growth requirements that make cross contamination a potential problem. Cell line contamination is not an uncommon incident in laboratories handling more than one cell line and many reports have been made on cross contamination of mammalian cell lines. Although problems of misidentification and cross-contamination of fish cell lines have rarely been reported, these are issues of concern for cell culturists that can make scientific results and their reproducibility unreliable. Proper identification of cell lines is thus crucial and protocols for routine and rapid screening are preferred. Cytogenetic evaluation, DNA fingerprinting, microsatellite analysis and PCR methods have been published for inter-species identification of many cell lines, but discerning intra-species contamination has been challenging. More complex DNA fingerprinting and hybridization techniques coupled with isoenzyme analysis have been developed to discriminate intra-species contamination, however, these require complex and time consuming procedures to enable cell identification thus are difficult to apply for routine use. A simple proteomic approach has been made to identify several fish cell lines derived from tissues of the same or differing species. Protein expression signatures (PES) of the evaluated fish cell lines have been developed using 2-DE and image analysis. A higher degree of concordance was seen among cell lines derived from rainbow trout, than from other fish species. Similar concordance was seen in cells derived from the same tissues than from other tissues within the same species. These profiles have been saved in an electronic databank and could be made available to be used for discerning the origins of the various cell lines evaluated. This proteomic approach could thus serve as an additional, valuable and reliable technique for the identification of fish cell lines.     

Antibody variable-region sequencing as a method for hybridoma cell-line authentication

Cross-contamination and misidentification of various cell lines is a widespread problem that can lead to spurious scientific conclusions. DNA fingerprinting is a powerful identification technique, which can be effectively used for the authentication of human cell lines. In contrast to human cancer cell lines, little attention has so far been given to establishing authentication practices for hybridoma cell lines. Since the majority of hybridomas stem from inbred animals, they have high genetic uniformity, which reduces the applicability of DNA fingerprinting. In the present study, we propose antibody variable-region sequencing as a method of choice for hybridoma cell-line authentication. This method focuses on the most diverse characteristic of hybridoma cell lines and thereby achieves a very high discriminatory power. The sequencing of light-chain variable regions has proven to be especially suitable for routine use because of its high success rate. Two other possible authentication methods, random amplified polymorphic DNA analysis and two-dimensional gel electrophoresis, were also examined. Compared to these and other methods that can be used for discrimination between hybridoma cell lines, variable-region sequencing has many advantages, most notably those of a very high discriminatory power, insensitivity to changes in experimental conditions, simple data analysis, and accessibility to most laboratories. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. Simon Koren and Miha Kosmač contributed equally to this work.     

中国驼鹿种群数量及分布现状的研究

1985一1987年间,作者在中国东北大、小兴安岭林区,根据三阶抽样的原理,选择1 6块样地,设置并调查202条样带,长2476．6km,遇见309条驼鹿足迹链。由此,明确了驼鹿在中国的分布区域并确定总栖息面积为1 9万多km² ．平均分布密度为0.0519头／km²,种群数量为9955士397头(a=0．2 ）,其种群下降率年平均逃6.3 %,且呈继续下降趋势, 亟待保护。     

CONSUMER DRIVEN POLLEN LIMITATION OF SEED PRODUCTION IN MARSH GRASSES

While flower predators can limit the sexual expression and seed production of salt marsh grasses, the relationship between these two effects of consumers has not been explored. At our study site, predation on Spartina patens, Spartina alterniflora, and Distichlis spicata was twice as high in 1985 (~70% ovule destruction) as in 1986 (~35% ovule destruction). In both years consumers destroyed flowers before maturity, reducing sexual expression, and particularly suppressed male sexual expression. Sexual suppression of males was much more pronounced in 1985 when flower predation was severe and the seed production of undamaged ovules was dramatically reduced. A number of lines of evidence suggest that predator limitation of male sexual expression and pollen supply contributed to low seed output in 1985. 1) Undamaged ovules of all three grasses protected from consumers but exposed to ambient windbome pollen set many more seeds in 1986 than in 1985, suggesting that pollen was more abundant in 1986; 2) Artificial pollinations revealed that marsh grasses are generally pollen-limited and that pollen limitation at our study site was more severe in 1985 than 1986; and 3) Caging stands of marsh grasses generally led to less predator damage, increased male densities and seed sets similar to those for hand-pollinated flowers. Our results support the hypothesis that flower predators can indirectly limit seed production by decreasing pollen availability.     

Utility of a C57BL/6 ES line versus 129 ES lines for Targeted Mutations in Mice

Inbred ES lines, though useful for generating targeted mutations in mice, are used infrequently. To appreciate the relative efficiency of inbred ES lines, a C57BL/6 ES line was compared with 129 strain ES lines for effectiveness in chimera formation leading to the establishment of targeted mutations in mice. Data from a transgenic facility spanning 7 years were collected. C57BL/6 ES cells injected into Balb/c embryos results in lower coat color chimerism than do 129 ES cells injected into C57BL/6 embryos. Combined data indicate that five independent targeted C57BL/6 clones should be injected as compared to three independent 129 clones to generate enough chimeras to effectively test for germ-line transmission. Thus, although less efficient than 129 ES lines, the C57BL/6 ES line is a relatively competent line and useful for the routine generation of targeted mutations in mice on a defined genetic background.     

Functional expression of dihydropyridine-insensitive calcium channels during PC12 cell differentiation by nerve growth factor (NGF), oncogenic ras,or src tyrosine kinase

1. Recombinant retroviruses were used to introduce a temperature-sensitive v-src gene and oncogenic c-Ha-ras into PC12 cells, and stable cell lines expressing these genes were established. 2. As previously reported, expression of v-src (Alema et al., 1985) or c-Ha-ras (Noda et al., 1985) in PC12 cells results in neurite outgrowth resembling that induced by NGF. We report here that v-src but not oncogenic c-Ha-ras induces a stable morphologic neuronal differentiation similar to treatment with NGF. Oncogenic c-Ha-ras-induced neurite outgrowth is not stable with long-term culture, rather the cells revert to an undifferentiated morphology with altered cell cycle kinetics. 3. The stable neuronal phenotype induced by v-src and NGF is characterized by the functional expression of dihydropyridine-insensitive calcium currents.     

Tick cell lines: tools for tick and tick-borne disease research

Over 40 cell lines are currently available from 13 ixodid and one argasid tick species. The successful isolation and propagation of several economically important tick-borne pathogens in tick cell lines has created a useful model to study interactions between tick cells and these viral and bacterial disease agents. Tick cell lines have already proved to be a useful tool in helping to define the complex nature of the host-vector-pathogen relationship. With the availability of genomics tools, tick cell lines will become increasingly important as a complement to tick and tick-borne disease research in vivo once genetic transformation and gene silencing using RNA interference become routine.     

Isoenzyme analysis as a rapid method for the examination of the species identity of cell cultures

Summary One of the major problems in cell culturing is the misidentification or cross-contamination of authentic continuous cell lines. We applied a rapid and efficient isoelectric focusing (IEF) technique for the routine analysis to detect interspecies contamination of cell cultures and for the identification of unknown animal cell lines. The method is based on the isoelectric separation of a specific set of intracellular enzymes which can be used to distinguish between cell lines of human, murine, or other mammalian origin. By means of preformed agarose gels, standardized conditions and equipment, this technique is especially applicable for routine work and allows the analysis of a large number of unknown samples with reproducible results. One hundred seventy-seven cell lines which have been sent to the Department of Human and Animal Cell Cultures at the DSM (Deutsche Sammlung von Mikroorganismen and Zellkulturen) were analyzed for species authentication; only three cell lines were found not to be of the presumed species. Our study strongly emphasizes standardized IEF as an efficient and rapid method for routinely monitoring the authenticity of cell lines.     

Social class differences in infant mortality in Sweden: comparison with England and Wales.

OBJECTIVES--To investigate social class differences in infant mortality in Sweden in the mid-1980s and to compare their magnitude with that of those found in England and Wales. DESIGN--Analysis of risk of infant death by social class in aggregated routine data for the mid-1980s, which included the linkage of Swedish births to the 1985 census. SETTING--Sweden and England and Wales. SUBJECTS--All live births in Sweden (1985-6) and England and Wales (1983-5) and corresponding infant deaths were analysed. The Swedish data were coded to the British registrar general''s social class schema. MAIN OUTCOME MEASURES--Risk of death in the neonatal and postneonatal period. RESULTS--Taking the non-manual classes as the reference group, in the neonatal period in Sweden the manual social classes had a relative risk for mortality of 1.20 (95% confidence interval 1.02 to 1.43) and those not classified into a social class a relative risk of 1.08 (0.88 to 1.33). In the postneonatal period the equivalent relative risks were 1.38 (1.08 to 1.77) for manual classes and 2.14 (1.65 to 2.79) for the residual; these are similar to those for England and Wales (1.43 (1.36 to 1.51) for manual classes, 2.62 (2.45 to 2.81) for the residual). CONCLUSIONS--The existence of an equitable health care system and a strong social welfare policy in Sweden has not eliminated inequalities in post-neonatal mortality. Furthermore, the very low risk of infant death in the Swedish non-manual group (4.8/1000 live births) represents a target towards which public health interventions should aim. If this rate prevailed in England and Wales, 63% of postneonatal deaths would be avoided.     

Predicting differentiation potential of human pluripotent stem cells:Possibilities and challenges

The capability of human pluripotent stem cell(hPSC) lines to propagate indefinitely and differentiate into derivatives of three embryonic germ layers makes these cells be powerful tools for basic scientific research and promising agents for translational medicine. However, variations in differentiation tendency and efficiency as well as pluripotency maintenance necessitate the selection of hPSC lines for the intended applications to save time and cost. To screen the qualified cell lines and exclude problematic cell lines, their pluripotency must be confirmed initially by traditional methods such as teratoma formation or by highthroughput gene expression profiling assay. Additionally, their differentiation potential, particularly the lineage-specific differentiation propensities of hPSC lines, should be predicted in an early stage. As a complement to the teratoma assay, RNA sequencing data provide a quantitative estimate of the differentiation ability of hPSCs in vivo. Moreover, multiple scorecards have been developed based on selected gene sets for predicting the differentiation potential into three germ layers or the desired cell type many days before terminal differentiation.For clinical application of hPSCs, the malignant potential of the cells must also be evaluated. A combination of histologic examination of teratoma with quantitation of gene expression data derived from teratoma tissue provides safety-related predictive information by detecting immature teratomas, malignancy marker expression, and other parameters. Although various prediction methods are available, distinct limitations remain such as the discordance of results between different assays and requirement of a long time and high labor and cost,restricting their wide applications in routine studies. Therefore, simpler and more rapid detection assays with high specificity and sensitivity that can be used to monitor the status of hPSCs at any time and fewer targeted markers that are more specific for a given desired cell type are urgently needed.