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Multilevel modelling of clustered grouped survival data using Cox regression model: an application to ART dental restorations.

In some controlled clinical trials in dental research, multiple failure time data from the same patient are frequently observed that result in clustered multiple failure time. Moreover, the treatments are often delivered by more than one operator and thus the multiple failure times are clustered according to a multilevel structure when the operator effects are assumed to be random. In practice, it is often too expensive or even impossible to monitor the study subjects continuously, but they are examined periodically at some regular pre-scheduled visits. Hence, discrete or grouped clustered failure time data are collected. The aim of this paper is to illustrate the use of the Monte Carlo Markov chain (MCMC) approach and non-informative prior in a Bayesian framework to mimic the maximum likelihood (ML) estimation in a frequentist approach in multilevel modelling of clustered grouped survival data. A three-level model with additive variance components model for the random effects is considered in this paper. Both the grouped proportional hazards model and the dynamic logistic regression model are used. The approximate intra-cluster correlation of the log failure times can be estimated when the grouped proportional hazards model is used. The statistical package WinBUGS is adopted to estimate the parameter of interest based on the MCMC method. The models and method are applied to a data set obtained from a prospective clinical study on a cohort of Chinese school children that atraumatic restorative treatment (ART) restorations were placed on permanent teeth with carious lesions. Altogether 284 ART restorations were placed by five dentists and clinical status of the ART restorations was evaluated annually for 6 years after placement, thus clustered grouped failure times of the restorations were recorded. Results based on the grouped proportional hazards model revealed that clustering effect among the log failure times of the different restorations from the same child was fairly strong (corr(child)=0.55) but the effects attributed to the dentists could be regarded as negligible (corr(dentist)=0.03). Gender and the location of the restoration were found to have no effects on the failure times and no difference in failure times was found between small restorations placed on molars and non-molars. Large restorations placed on molars were found to have shorter failure times compared to small restorations. The estimates of the baseline parameters were increasing indicating increasing hazard rates from interval 1 to 6. Results based on the logistic regression models were similar. In conclusion, the use of the MCMC approach and non-informative prior in a Bayesian framework to mimic the ML estimation in a frequentist approach in multilevel modelling of clustered grouped survival data can be easily applied with the use of the software WinBUGS.

Adolescent↗

An intrinsic neuronal-like network in the rat pineal gland.

Recent studies have shown that in rat pineal glands kept in vitro action potential-producing cell clusters are demonstrable. To test whether the clusters interact, multiple-unit recordings were carried out simultaneously from different clusters, with or without electrical stimulation. Clusters with rhythmic burst activity exhibit highly synchronized firing and electrical stimulation of one cluster elicits an immediate response in another one, apparently involving synapses but not gap junctions. It is hypothesized that the interacting clusters form a network. As the firing is affected by norepinephrine, acetylcholine and Ca2+, the network may monitor the interstitial concentrations of these substances and spread this information in the gland, to modulate melatonin secretion.

Animals↗

Microarray and comparative genomics-based identification of genes and gene regulatory regions of the mouse immune system.

BACKGROUND: In this study we have built and mined a gene expression database composed of 65 diverse mouse tissues for genes preferentially expressed in immune tissues and cell types. Using expression pattern criteria, we identified 360 genes with preferential expression in thymus, spleen, peripheral blood mononuclear cells, lymph nodes (unstimulated or stimulated), or in vitro activated T-cells. RESULTS: Gene clusters, formed based on similarity of expression-pattern across either all tissues or the immune tissues only, had highly significant associations both with immunological processes such as chemokine-mediated response, antigen processing, receptor-related signal transduction, and transcriptional regulation, and also with more general processes such as replication and cell cycle control. Within-cluster gene correlations implicated known associations of known genes, as well as immune process-related roles for poorly described genes. To characterize regulatory mechanisms and cis-elements of genes with similar patterns of expression, we used a new version of a comparative genomics-based cis-element analysis tool to identify clusters of cis-elements with compositional similarity among multiple genes. Several clusters contained genes that shared 5-6 cis-elements that included ETS and zinc-finger binding sites. cis-Elements AP2 EGRF ETSF MAZF SP1F ZF5F and AREB ETSF MZF1 PAX5 STAT were shared in a thymus-expressed set; AP4R E2FF EBOX ETSF MAZF SP1F ZF5F and CREB E2FF MAZF PCAT SP1F STAT cis-clusters occurred in activated T-cells; CEBP CREB NFKB SORY and GATA NKXH OCT1 RBIT occurred in stimulated lymph nodes. CONCLUSION: This study demonstrates a series of analytic approaches that have allowed the implication of genes and regulatory elements that participate in the differentiation, maintenance, and function of the immune system. Polymorphism or mutation of these could adversely impact immune system functions.

Animals↗

The cluster phenomenon in patients who have multiple vertebral compression fractures.

Little is known about the natural history of patients who have multiple compression fractures. During an eight-year period, eight patients who suffered relentless nonmechanical back pain after an initial compression fracture were evaluated. Radiographs and radionuclide bone scans were obtained in all eight patients and showed the temporal clustering of at least five spontaneous thoracic or lumbar vertebral compression fractures within a period of eight months. The patient database during this eight-year period included 21 patients with a similar number of fractures but without the phenomenon of temporal clustering. Age and gender distribution, premorbid activity levels, presence of secondary causes of osteopenia, dietary history, and vertebral bone density measurements were similar in both groups. Five of the eight patients with cluster fractures were on chronic glucocorticoid therapy for treatment of an underlying illness. All five patients experienced an exacerbation of their chronic illness that required an increase in the dose of their steroid medication within three months preceding the onset of the cluster fractures. No patient in the noncluster fracture control group experienced a similar exacerbation of an underlying illness or required a similar boost in their steroid medication. The temporal clustering of numerous fractures (cluster phenomenon) is a cause of transient severe disability in patients who have profound osteopenia, and may be precipitated by exacerbation of an intercurrent illness or by an associated obligate increase in glucocorticoid medication in a patient with preexisting severe osteopenia.

Aged↗

Relationship between the degree of individual space-time clustering and age at onset of disease among multiple sclerosis patients.

A previous study of 381 multiple sclerosis (MS) patients from Norway has shown significant clustering of residence in adolescence among contemporaneous patients. Using a regression model with individual degree of clustering determined by Monte Carlo methods, this study utilizes the same data and shows a significant correlation (P = 0.002) between the degree of clustering and age at clinical onset of disease. A high degree of clustering in adolescence indicated a low age at clinical onset. This association was also verified independently of the regression model. The finding supports the theory of a common viral agent acting epidemically among nonpre-exposed individuals during adolescence, with MS developing in individuals with other necessary factors, such as a genetic predisposition.

Adolescent↗

Multiple sclerosis distribution in northern Sardinia: spatial cluster analysis of prevalence.

BACKGROUND: A heterogeneous geographic distribution of MS has been reported among different ethnic groups, and also within small communities. Epidemiologic studies conducted over the past two decades using repeated assessments clearly show that Sardinia is at high risk for MS, with a prevalence of 150 per 100,000 in 1997. OBJECTIVE: To present spatial analysis of the disease prevalence to disclose possible "hot" or "cold" spots of disease, further allowing correlations with risk factors. METHODS: A spatial analysis of the whole province of Sassari, in northern Sardinia, at a microgeographic level (i.e., in the 89 administrative communes and 6 linguistic areas) was conducted. Because of the small number of cases per commune and to overcome random variability, a hierarchical Bayesian approach was adopted. The distribution of prevalent cases by commune of residence on December 31, 1997 and from age 5 to 15 years was analyzed. RESULTS: A clustering pattern was found in the southwestern communes of the province based on geographic distribution by both prevalence and residence at age 5 to 15 years. A west-to-east gradient also was observed. CONCLUSIONS: This study highlights a hot spot of MS in the southwestern part of Sassari province, bordering with the commune of Macomer, where MS was once hypothesized as having occurred as an epidemic. Interestingly, these areas of MS clustering comprise the Common Logudorese linguistic domain. The Catalan area, linguistically and genetically distant from the remaining Sardinian domains, does not show such high estimates. Because MS is not a single-source infectious disease, this study may help test the hypothesis that a widely and evenly spread environmental (infectious?) agent may produce disease in subgroups of genetically more susceptible individuals in areas at higher inbreeding rates, wherein a disease mode of inheritance could be better investigated.

Adolescent↗

Large-scale sequencing of the CD33-related Siglec gene cluster in five mammalian species reveals rapid evolution by multiple mechanisms.

Siglecs are a recently discovered family of animal lectins that belong to the Ig superfamily and recognize sialic acids (Sias). CD33-related Siglecs (CD33rSiglecs) are a subgroup with as-yet-unknown functions, characterized by sequence homology, expression on innate immune cells, conserved cytosolic tyrosine-based signaling motifs, and a clustered localization of their genes. To better understand the biology and evolution of CD33rSiglecs, we sequenced and compared the CD33rSiglec gene cluster from multiple mammalian species. Within the sequenced region, the segments containing CD33rSiglec genes showed a lower degree of sequence conservation. In contrast to the adjacent conserved kallikrein-like genes, the CD33rSiglec genes showed extensive species differences, including expansions of gene subsets; gene deletions, including one human-specific loss of a novel functional primate Siglec (Siglec-13); exon shuffling, generating hybrid genes; accelerated accumulation of nonsynonymous substitutions in the Sia-recognition domain; and multiple instances of mutations of an arginine residue essential for Sia recognition in otherwise intact Siglecs. Nonsynonymous differences between human and chimpanzee orthologs showed uneven distribution between the two beta sheets of the Sia-recognition domain, suggesting biased mutation accumulation. These data indicate that CD33rSiglec genes are undergoing rapid evolution via multiple genetic mechanisms, possibly due to an evolutionary "arms race" between hosts and pathogens involving Sia recognition. These studies, which reflect one of the most complete comparative sequence analyses of a rapidly evolving gene cluster, provide a clearer picture of the ortholog status of CD33rSiglecs among primates and rodents and also facilitate rational recommendations regarding their nomenclature.

Animals↗

Single-step co-integration of multiple expressible heterologous genes into the ribosomal DNA of the methylotrophic yeast Hansenula polymorpha.

We have investigated the methylotrophic yeast Hansenula polymorpha as a host for the co-integration and expression of multiple heterologous genes using an rDNA integration approach. The ribosomal DNA (rDNA) of H. polymorpha was found to consist of a single rDNA cluster of about 50-60 repeats of an 8-kb unit located on chromosome II. A 2.4-kb segment of H. polymorpha rDNA encompassing parts of the 25S, the complete 5S and the non-transcribed spacer region between 25S and 18S rDNA was isolated and inserted into conventional integrative H. polymorpha plasmids harboring the Saccharomyces- cerevisiae-derived URA3 gene for selection. These rDNA plasmids integrated homologously into the rDNA repeats of a H. polymorpha (odc1) host as several independent clusters. Anticipating that this mode of multiple-cluster integration could be used for the simultaneous integration of several distinct rDNA plasmids, the host strain was co-transformed with a mixture of up to three different plasmids, all bearing the same URA3 selection marker. Transformations indeed resulted in mitotically stable strains harboring one, two, or all three plasmids integrated into the rDNA. The overall copy number of the plasmids integrated did not exceed the number of rDNA repeats present in the untransformed host strain, irrespective of the number of different plasmids involved. Strains harboring different plasmids co-expressed the introduced genes, resulting in functional proteins. Thus, this approach provides a new and attractive tool for the rapid generation of recombinant strains that simultaneously co-produce several proteins in desired stoichiometric ratios.

Base Sequence↗

Co-contribution of IP3R and Ca2+ influx pathways to pacemaker Ca2+ activity in stomach ICC.

Intracellular Ca2+ oscillations in interstitial cells of Cajal (ICCs) are thought to be the primary pacemaker activity in the gut. In the present study, the authors prepared small tissues of 100-to 300-microm diameter (cell cluster preparation) from the stomach smooth muscle (including the myenteric plexus) of mice by enzymatic and mechanical treatments. After 2 to 4 days of culture, the intracellular Ca2+ concentration ([Ca2+]i) was measured. In the presence of nifedipine, a dihydropyridine Ca2+ channel antagonist, spontaneous [Ca2+]i oscillations were observed within limited regions showing positive c-Kitimmunoreactivity, a maker for ICCs. In the majority of cell cluster preparations with multiple regions of [Ca2+]i oscillations, [Ca2+]i oscillated synchronously in the same phase. A small number of cell clusters (8 of 53) showed multiple regions of [Ca2+]i oscillations synchronized but with a considerable phase shift. Neither tetrodotoxin (250 nM) nor atropine (10 microM) significantly affected [Ca2+]i oscillations in the presence of nifedipine. Low concentrations (40 microM) of Ni2+ had little effect on the spontaneous [Ca2+]i oscillation, but SK&F96365 (40 microM) and Cd2+ (120 microM) terminated it. Applications of either 2-aminoethoxydiphenyl borate (10 microM) or xestosponginC(10 microM) completely and rather rapidly (approximately 2 min) abolished the spontaneous [Ca2+]i oscillations. The results suggest that pacemaker [Ca2+]i oscillations in ICCs are produced by close interaction of intracellular Ca2+ release channels, especially inositol 1,4,5-trisphosphate receptor (InsP3R) and Ca2+ influx pathways, presumably corresponding to store-operated type channels. Reverse transcription polymerase chain reaction examinations revealed expression of TRPC2, 4, and 6, as well as InsP3R1 and 2 in ICCs.

Animals↗

Functional outcomes in self-care, mobility, communication, and learning in extremely low-birth weight infants.

Gaps have existed in specifying degrees of severity of cerebral palsy assessment of self-care and communicative competencies, and specifying age-appropriate preschool educational and behavioral competencies. Imbedded in the concept of measuring functional status is the interaction between health and neurologic impairments, developmental challenges and competencies, family resources and disadvantages, and the child's current status. In reviewing historic outcomes of severe ROP over the past 40 years, it was noted that severe ROP caused blindness in 2% to 11% of survivors. There was a constant observation that approximately 50% of severe ROP survivors with blindness had multiple functional and developmental challenges beyond blindness alone. Similarly, in reviewing outcomes of cerebral palsy, it is imperative to describe the severity of cerebral palsy and functional consequences in motor, selfcare, communication, and learning. The reason to measure the functional status of children with neurodevelopmental impairments before first grade is that the degrees of severity of these disorders can be specified before attending school with peers. Subtler aspects of neurodevelopmental impairments need to assess impact on literacy, information learning, written language, social competencies with peers, and recreational and community participation. In this way, we can understand the vulnerabilities and resiliences of children and families of VLBW and ELBW status. This is a critical step in understanding long-term quality of life and independent living issues. In addition, our efforts can address those factors and pathways whereby multiple disabilities and multiple functional limitations cluster. Our biomedical intervention can prioritize strategies that lessen severe multiple disabilities and simultaneously support families, when despite our best efforts functional challenges are life long.

Activities of Daily Living↗

Clustering of Candida infections in the neonatal intensive care unit: concurrent emergence of multiple strains simulating intermittent outbreaks.

BACKGROUND: Clusters of Candida albicans and Candida parapsilosis infections were noted intermittently in our neonatal intensive care unit (NICU). We attempted to determine whether these clusters represented single strain outbreaks or coincidental emergence of unrelated strains. METHODS: A retrospective examination of the frequency of candidemia during a 9-year period, two point prevalence studies of colonization and assessment of strain relatedness of individual infant isolates during and in between clusters during a 2-year period with karyotyping and restriction endonuclease analysis of genomic DNA (REAG). RESULTS: C. albicans and C. parapsilosis infections emerged in a scattered pattern (1 to 2 cases every few months) with intermittent clustering of 3 cases/month. The colonization rate was 50% 5 weeks after an apparent cluster, equally distributed between C. albicans and C. parapsilosis, and 17.6% (exclusively with C. parapsilosis) 4 months after absence of invasive disease. Utilizing REAG or karyotyping singly we noted 12 and 16 DNA banding patterns, respectively, among 23 infant isolates. Few patterns were observed repeatedly over 2- to 20-month periods, implying recurrent emergence of the same strains. Combining karyotyping with REAG revealed a different epidemiologic pattern. It identified 20 distinct composites with identical composites in 3 infant pairs. All infants with identical composites were in the NICU concurrently. The frequency of strain relatedness was comparable among clustered cases (16.7%), scattered cases (7.7%) and simultaneously colonized infants (16.7%). CONCLUSIONS: These findings illustrate that Candida infections clustered periodically in our NICU but that these clusters were often caused by unrelated strains with infrequent cross-infection during and between clusters. With suboptimal typing this pattern of emergence can be mistaken for same strain outbreaks.

Candida↗

Multiple aromaticity and antiaromaticity in silicon clusters.

A series of silicon clusters containing four atoms but with different charge states (Si4(2+), Si4, Si4(2-), and NaSi4-) were studied by photoelectron spectroscopy and ab initio calculations. Structure evolution and chemical bonding in this series were interpreted in terms of aromaticity and antiaromaticity, which allowed the prediction of how structures of the four-atom silicon clusters change upon addition or removal of two electrons. It is shown that Si4(2+) is square-planar, analogous to the recently discovered aromatic Al4(2-) cluster. Upon of two electrons, neutral Si4 becomes sigma-antiaromatic and exhibits a rhombus distortion. Adding two more electrons to Si4 leads to two energetically close structures of Si4(2-): either a double antiaromatic parallelogram structure or an aromatic system with a butterfly distortion. Because of the electronic instability of doubly charged Si4(2-), a stabilizing cation (Na+) was used to produce Si4(2-) in the gas phase in the form of Na+[Si4(2-)], which was characterized experimentally by photoelectron spectroscopy. Multiple antiaromaticity in the parallelogram Na+[Si4(2-)] species is highly unusual.

Journal Article↗

Microswitch clusters promote adaptive responses and reduce finger mouthing in a boy with multiple disabilities.

The authors assessed new microswitch clusters (i.e., combinations of two microswitches) and contingent stimulation to increase adaptive responses (i.e., foot and head movements) and reduce aberrant behavior (i.e., finger mouthing)in a boy with multiple disabilities. Initially, intervention was directed at increasing the frequency of each adaptive response, individually, through contingent use of preferred stimuli. Subsequently, adaptive responses led to preferred stimuli only if they occurred in the absence of finger mouthing. Postintervention checks occurred 1, 2, and 3 months after the end of the intervention. Data showed that the boy (a) increased the frequency of the adaptive responses extensively, (b) learned to perform these responses largely free from finger mouthing, and (c) maintained these changes over time. The importance of microswitch clusters to simultaneously pursue the dual objective of promoting adaptive responses and reducing aberrant ones is underlined.

Abnormalities, Multiple↗

Use of artificial genomes in assessing methods for atypical gene detection.

Parametric methods for identifying laterally transferred genes exploit the directional mutational biases unique to each genome. Yet the development of new, more robust methods--as well as the evaluation and proper implementation of existing methods--relies on an arbitrary assessment of performance using real genomes, where the evolutionary histories of genes are not known. We have used the framework of a generalized hidden Markov model to create artificial genomes modeled after genuine genomes. To model a genome, "core" genes--those displaying patterns of mutational biases shared among large numbers of genes--are identified by a novel gene clustering approach based on the Akaike information criterion. Gene models derived from multiple "core" gene clusters are used to generate an artificial genome that models the properties of a genuine genome. Chimeric artificial genomes--representing those having experienced lateral gene transfer--were created by combining genes from multiple artificial genomes, and the performance of the parametric methods for identifying "atypical" genes was assessed directly. We found that a hidden Markov model that included multiple gene models, each trained on sets of genes representing the range of genotypic variability within a genome, could produce artificial genomes that mimicked the properties of genuine genomes. Moreover, different methods for detecting foreign genes performed differently--i.e., they had different sets of strengths and weaknesses--when identifying atypical genes within chimeric artificial genomes.

Algorithms↗

Marginal regression of multivariate event times based on linear transformation models.

Multivariate event time data are common in medical studies and have received much attention recently. In such data, each study subject may potentially experience several types of events or recurrences of the same type of event, or event times may be clustered. Marginal distributions are specified for the multivariate event times in multiple events and clustered events data, and for the gap times in recurrent events data, using the semiparametric linear transformation models while leaving the dependence structures for related events unspecified. We propose several estimating equations for simultaneous estimation of the regression parameters and the transformation function. It is shown that the resulting regression estimators are asymptotically normal, with variance-covariance matrix that has a closed form and can be consistently estimated by the usual plug-in method. Simulation studies show that the proposed approach is appropriate for practical use. An application to the well-known bladder cancer tumor recurrences data is also given to illustrate the methodology.

Biomedical Research↗

Obsessive-compulsive disorder subgroups: a symptom-based clustering approach.

Although obsessive-compulsive disorder (OCD) is often considered a heterogeneous condition, there is no generally accepted subtype typology. Cluster analysis was used to identify definitive symptom-based groupings of 106 OCD patients. A stable cluster solution was achieved and five patient subgroups were identified based on their pattern of symptoms on the Yale-Brown (Y-BOCS) symptom checklist: harming, hoarding, contamination, certainty and obsessionals. The five subgroups were characterized by dominant symptom patterns and significant secondary concerns reflecting the symptom heterorgenaity often seen in the clinical presentation of obsessional patents. Between cluster differences on multiple symptom measures were evaluated and several meaningful differences were identified. Cluster analytic procedures may prove to be a useful tool for identifying a functional taxonomy of OCD subtypes.

Adult↗

MIL-50, an open-framework GaPO with a periodic pattern of small water ponds and dry rubidium atoms: a combined XRD, NMR, and computational study.

A new fluorinated gallium phosphate, MIL-50, has been synthesized under mild hydrothermal conditions using 1,6-diaminohexane. The chemical formula of MIL-50 is Rb(2)Ga(9)(PO(4))(8)(HPO(4))(OH)F(6).2N(2)C(6)H(18).7H(2)O. The structure is a network of hexameric units of Ga(3)(PO(4))(3)F(2) and Ga(3)(PO(4))(2)(HPO(4))F(3) via corner sharing. It creates a three-dimensional open-framework delimiting 6- and 18-ring channels running along the c axis. The diprotonated 1,6-diaminohexane and water molecules are trapped within the 18-ring pores, whereas the rubidium cations reside in the 6-ring ones. A double quantum (31)P NMR experiment and partial charge calculations indicate that water molecules are present under the form of periodic small clusters, lowering the multiplicity of one phosphorus site, P3. Though water hops within the clusters, the motion leaves the water pattern periodic. Rubidium is so tightly embedded into the framework that water moving in the large 18-ring channels does not reach it, leaving it therefore dry. The crystal framework may be ascribed to the orthorhombic space group Cmc2(1) (n degrees 36), a = 32.1510(2), b = 17.2290(3), c = 10.2120(1) A. The periodic water pattern has a different symmetry than that of the framework. A method has been devised to superpose the two sublattices that coexist in the same unit cell in order to have full occupancy of each site and to perform Madelung summations. This original method is of general interest for most zeolitic materials exhibiting a different symmetry for the framework and the template sublattices.

Journal Article↗