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Cytokeratin expression in oral exfoliative cytology: effect of temperature and fixation.

The identification of keratin expression within oral cytology may be useful in the diagnosis of clinically suspicious oral mucosal lesions. There may be wide variation in the temperature at which such smears are stored, prior to processing. Conventionally, rapid fixation or storage at low temperatures is recommended to preserve kertin expression within tissue biopsies. No previous study has assessed whether this is true for oral cytology. Smears were taken from clinically normal buccal mucosa. For each temperature assessed (-70, -40, -22, +5, +20 and +26 degrees C), one smear was spray-fixed (Vale Smear Fix) and one air-dried, prior to storage for 4 days, and then staining with the pan-epithelial antikeratin antibody, LP34. Preservation of keratin expression was assessed as either weak (zero to few positive cells) or strong (most cells positive). The results were analysed using logistic regression with the statistical modelling package, GLIM. Over the range of temperatures studied, spray fixation did not appear to improve the identification of keratin expression. Although the best preservation was obtained at lower temperatures, keratin expression was still adequate after 4 days at 20 degrees C. Hence, a delay in processing of 4 days would still allow detectable expression in oral exfoliative cytology even at room temperature.

Adolescent↗

New variants in the CACNA1H gene identified in childhood absence epilepsy.

Childhood absence epilepsy (CAE) is a common form of idiopathic generalized epilepsy with polygenic inheritance. In our previous studies, relatively high frequent variants in the T-type calcium channel gene, CACNA1H, were identified in the Chinese Han population, most of which are located in exons 6-12. The goal of this study was to identify additional variants in this region of the CACNA1H gene. To this end, exons 6-12 were sequenced in 100 newly recruited CAE trios and 191 normal controls. Thirty-nine variants were identified in CAE trios or controls, 14 of which were found only in CAE patients, including two nonsynonymous variants that were newly found. Thirteen of the 39 variants were found in both CAE patients and controls, 11 were found only in parents of CAE trios, and one was found only in controls. Twenty-eight of these variants had not been previously reported. Both permutation test and transmission/disequilibrium test (TDT) indicated that a SNP-52037C>T in intron11 was significant in association with CAE. In conclusion, these data further support the hypothesis that CACNA1H is an important susceptibility gene for CAE in the Chinese Han population.

Asian People↗

Hypothesis for the causes and periodicity of repetitive linear enamel hypoplasia in large, wild African (Pan troglodytes and Gorilla gorilla) and Asian (Pongo pygmaeus) apes.

Repetitive linear enamel hypoplasia (rLEH) is often observed in recent large-bodied apes from Africa and Asia as well as Mid- to Late Miocene sites from Spain to China. The ubiquity and periodicity of rLEH are not understood. Its potential as an ontogenetic marker of developmental stress in threatened species (as well as their ancient relatives) makes rLEH an important if enigmatic problem. We report research designed to show the periodicity of rLEH among West African Pan troglodytes (12 male, 32 female), Gorilla gorilla (10 male, 10 female), and Bornean and Sumatran Pongo pygmaeus (11 male, 9 female, 9 unknown) from collections in Europe. Two methods were employed. In the common chimpanzees and gorillas, the space between adjacent, macroscopically visible LEH grooves on teeth with two or more episodes was expressed as an absolute measure and as a ratio of complete unworn crown height. In the orangutans, the number of perikymata between episode onsets, as well as duration of rLEH, was determined from scanning electron micrographs of casts of incisors and canines. We conclude that stress in the form of LEH commences as early as 2.5 years of age in all taxa and lasts for several years, and even longer in orangutans; the stress is not chronic but episodic; the stressor has a strong tendency to occur in pulses of two occurrences each; and large apes from both land masses exhibit rLEH with an average periodicity of 6 months (or multiples thereof; Sumatran orangutans seem to show only annual stress), but this needs further research. This is supported by evidence of spacing between rLEH as well as perikymata counts. Duration of stress in orangutans averages about 6 weeks. Finally, the semiannual stressor transcends geographic and temporal boundaries, and is attributed to regular moisture cycles associated with the intertropical convergence zone modified by the monsoon. While seasonal cycles can influence both disease and nutritional stress, it is likely the combination of seasonal variation in fruiting cycles with specific stressors (malaria and/or intestinal parasites, especially hookworm) that results in this widespread phenomenon. This seasonal stress is sufficiently common and of long duration (6 weeks on average in orangutans) that we think rLEH may reflect significant stress in recent and, inferentially, fossil apes. Increasing seasonality may have impinged negatively on later Miocene apes, especially if they lacked a clear birth peak or seasonality in their reproductive cycles.

Animals↗

Correction of through-plane deformation artifacts in stimulated echo acquisition mode cardiac imaging.

Attempts to use a stimulated echo acquisition mode (STEAM) in cardiac imaging are impeded by imaging artifacts that result in signal attenuation and nulling of the cardiac tissue. In this work, we present a method to reduce this artifact by acquiring two sets of stimulated echo images with two different demodulations. The resulting two images are combined to recover the signal loss and weighted to compensate for possible deformation-dependent intensity variation. Numerical simulations were used to validate the theory. Also, the proposed correction method was applied to in vivo imaging of normal volunteers (n = 6) and animal models with induced infarction (n = 3). The results show the ability of the method to recover the lost myocardial signal and generate artifact-free black-blood cardiac images.

Animals↗

Distinctive distribution of AIM1 polymorphism among major human populations with different skin color.

The genetic background for human skin color has been a major topic in human genetics; however, its molecular basis is still unclear. The gene for the AIM-1 protein (AIM1) was recently found to be responsible for the body color of medaka fish. In the search for the genes controlling human skin color variations, we have investigated genetic polymorphisms of this gene, and we have found a single-nucleotide polymorphism that has clear association with major human populations in terms of skin color.

Animals↗

Density-based load estimation using two-dimensional finite element models: a parametric study.

A parametric investigation was conducted to determine the effects on the load estimation method of varying: (1) the thickness of back-plates used in the two-dimensional finite element models of long bones, (2) the number of columns of nodes in the outer medial and lateral sections of the diaphysis to which the back-plate multipoint constraints are applied and (3) the region of bone used in the optimization procedure of the density-based load estimation technique. The study is performed using two-dimensional finite element models of the proximal femora of a chimpanzee, gorilla, lion and grizzly bear. It is shown that the density-based load estimation can be made more efficient and accurate by restricting the stimulus optimization region to the metaphysis/epiphysis. In addition, a simple method, based on the variation of diaphyseal cortical thickness, is developed for assigning the thickness to the back-plate. It is also shown that the number of columns of nodes used as multipoint constraints does not have a significant effect on the method.

Animals↗

A preliminary comparative analysis of primate segmental duplications shows elevated substitution rates and a great-ape expansion of intrachromosomal duplications.

Compared with other sequenced animal genomes, human segmental duplications appear larger, more interspersed, and disproportionately represented as high-sequence identity alignments. Global sequence divergence estimates of human duplications have suggested an expansion relatively recently during hominoid evolution. Based on primate comparative sequence analysis of 37 unique duplication-transition regions, we establish a molecular clock for their divergence that shows a significant increase in their effective substitution rate when compared with unique genomic sequence. Fluorescent in situ hybridization (FISH) analyses from 1053 random nonhuman primate BACs indicate that great-ape species have been enriched for interspersed segmental duplications compared with representative Old World and New World monkeys. These findings support computational analyses that show a 12-fold excess of recent (>98%) intrachromosomal duplications when compared with duplications between nonhomologous chromosomes. These architectural shifts in genomic structure and elevated substitution rates have important implications for the emergence of new genes, gene-expression differences, and structural variation among humans and great apes.

Animals↗

Divergence of the genes on human chromosome 21 between human and other hominoids and variation of substitution rates among transcription units.

The study of genomic divergence between humans and primates may provide insight into the origins of human beings and the genetic basis of unique human traits and diseases. Chromosome 21 is the smallest chromosome in the human genome, and some of its regions have been implicated in mental retardation and other diseases. In this study, we sequenced the coding and regulatory regions of 127 known genes on human chromosome 21 in DNA samples from human and chimpanzees and a part of the corresponding genes from orangutan, gorilla, and macaque. Overall, 3,003 nucleotide differences between human and chimpanzee were identified over approximately 400 kb. The differences in coding, promoter, and exon-intron junction regions were 0.51 +/- 0.02%, 0.88 +/- 0.03%, and 0.85 +/- 0.02%, respectively, much lower than the previously reported 1.23% in genomic regions, which suggests the presence of purifying selection. Significant variation in substitution rate among genes was observed by comparing the divergence between human and chimpanzee. Furthermore, by implementing a bioinformatics-based approach, we showed that the identification of genetic variants specific to the human lineage might lead to an understanding of the mechanisms that are attributable to the phenotypes that unique to humans, by changing the structure and/or dosage of the proteins expressed. A phylogenetic analysis unambiguously confirms the conclusion that chimpanzees were our closest relatives to the exclusion of other primates and the relative divergence of the Homo-Pan and that of (Homo-Pan)-Gorilla are 4.93 million years and 7.26 million years, respectively.

3' Untranslated Regions↗

[Macrophages in rheumatoid synovial membrane: an update].

The immunophenotype of lining and subintimal synovial mononuclear phagocytes (MP) of rheumatoid arthritis (RA) were sought by immunohistology and compared with osteoarthritis (OA) tissue in order to determine the significance of MPs in the pathobiology of RA. Almost all the lining cells (SLCs) in RA consisted of MPs (80 to 90% expressing CD45/CD14/CD68). A major proportion of the interaggregate areas of the rheumatoid subintima was also made of MP cells (50 to 70% expressing CD14/CD68). A marked variation in the immunohistological reaction of antibodies reacting within intimal MPs and between intimal and subintimal MPs was found. Intimal MPs expressed a wide range of macrophage-associated antigens, including receptors for Fc (CD16, CD32, CD64) and complement (CD35, CD11b, CD11c) as well as several integrin and non-integrin cell adhesion molecules (CD29/CD49b, CD49d, CD49f, CD51/CD61, CD11a, CD31, CD54, CD44, CD9, CD63). The monocyte marker, CD14, was down-regulated on SLCs in both RA and OA. When compared to intimal expression of leucocyte common antigen (CD45), CD68, a pan-macrophage maturation antigen, was found to be an unreliable macrophage antigen in OA intima. There was no difference in antigenic phenotype of SLCs in inflammatory and non-inflammatory OA with early activation markers (CD25, CD71) mainly present on MPs. In RA, synovial MPs showed increased expression of activation, maturation and functional antigens suggesting that they are rapidly and fully activated. The fact that their recruitment was independent of the degree of lymphocyte infiltration further emphasises the central importance of synovial MPs in RA.

Aged↗

Catarrhine primate divergence dates estimated from complete mitochondrial genomes: concordance with fossil and nuclear DNA evidence.

Accurate divergence date estimates improve scenarios of primate evolutionary history and aid in interpretation of the natural history of disease-causing agents. While molecule-based estimates of divergence dates of taxa within the superfamily Hominoidea (apes and humans) are common in the literature, few such estimates are available for the Cercopithecoidea (Old World monkeys), the sister taxon of the hominoids in the primate infraorder Catarrhini. To help fill this gap, we have sequenced the entire mitochondrial DNA (mtDNA) genomes from a representative of three cercopithecoid tribes, Cercopithecini (Chlorocebus aethiops), Colobini (Colobus guereza), and Presbytini (Trachypithecus obscurus), and analyzed these new data together with other catarrhine mtDNA genomes available in public databases. Molecular divergence date estimates are dependent on calibration points gleaned from the paleontological record. We defined criteria for the selection of good calibration points and identified three points meeting these criteria: Homo-Pan, 6.0 Ma; Pongo-hominines, 14.0 Ma; hominoid/cercopithecoid, 23.0 Ma. Because a uniform molecular clock does not fit the catarrhine mtDNA data, we estimated divergence dates using a penalized likelihood and a Bayesian method, both of which take into account the effects of rate differences on lineages, phylogenetic tree structure, and multiple calibration points. The penalized likelihood method applied to the coding regions of the mtDNA genome yielded the following divergence date estimates, with approximate 95% confidence intervals: cercopithecine-colobine, 16.2 (14.4-17.9) Ma; colobin-presbytin, 10.9 (9.6-12.3) Ma; cercopithecin-papionin, 11.6 (10.3-12.9) Ma; and Macaca-Papio, 9.8 (8.6-10.9) Ma. Within the hominoids, the following dates were inferred: hylobatid-hominid, 16.8 (15.0-18.5) Ma; Gorilla-Homo+Pan, 8.1 (7.1-9.0) Ma; Pongo pygmaeus pygmaeus-P. p. abelii, 4.1 (3.5-4.7) Ma; and Pan troglodytes-P. paniscus, 2.4 (2.0-2.7) Ma. These dates were similar to those found using penalized likelihood on other subsets of the data, but slightly younger than several of the Bayesian estimates.

Africa↗

Hemagglutination with crane herpesvirus.

A crane herpesvirus (CrHV) grown in chicken embryo fibroblast (CEF) cells was tested for hemagglutination (HA) with erythrocytes from a variety of species at 4 degrees C, room temperature and 37 degrees C. HA was observed at all temperatures with erythrocytes from mouse, ddY and BALB/c strains, but not with those from cattle, sheep and chicken. Mice, ddY strain, showed an individual variation in agglutinability of their erythrocytes and erythrocytes from BALB/c gave a higher HA titer. The HA activity was inhibited by the sera obtained from naturally infected cranes, experimentally infected duck and immunized rabbit with CrHV. HI antibody titers of these sera showed a closely positive correlation with their neutralizing antibody titers.

Animals↗

Quantifying the effect of IV contrast media on integrated PET/CT: clinical evaluation.

OBJECTIVES: The use of IV contrast media in PET/CT can result in an overestimation of PET attenuation factors that potentially can affect interpretation. The objective of this study was to quantify the effect of IV contrast media in PET/CT and assess its impact on patients with intrathoracic malignancies. MATERIALS AND METHODS: Nine patients had CTs performed with and without IV contrast media followed by (18)F-FDG PET. PET images were reconstructed using contrast-enhanced and unenhanced CT. To quantify the effect of contrast media on standardized uptake values (SUV), similar regions of interest (ROIs) were drawn on the subclavian vein, heart, liver, spleen, and site of malignancy on both CT and corresponding reconstructed PET images, and the mean and maximum values were compared. In addition, two physicians blinded to the imaging parameters that were used evaluated the reconstructed PET images to assess whether IV contrast media had an effect on clinical interpretation. RESULTS: For all patient studies, the subclavian vein region on the ipsilateral side of contrast media administration had the highest increase in CT numbers with a corresponding average SUV(max) increase of 27.1%. Similarly, ROIs of the heart and at the site of malignancy showed an increase in the maximum attenuation value with a corresponding average SUV(max) increase of 16.7% and 8.4%, respectively. Other locations had relatively small attenuation value differences with a correspondingly negligible SUV variation. CONCLUSION: Although there is a significant increase in SUV in regions of high-contrast concentration when contrast-enhanced CT is used for attenuation correction, this increase is clinically insignificant. Accordingly, in PET/CT, IV contrast-enhanced CT can be used in combination with the PET to evaluate patients with cancer.

Contrast Media↗

[Cloning to rule out 10 candidate genes located in chromosome 12q24 for Charcot-Marie-Tooth disease type 2L].

OBJECTIVE: To clone the disease-causing genes possibly existing in 6.8 cM distance between microsatellite markers D12S1720 and D12S1611 in chromosome 12q24 for Charcot-Marie-Tooth disease type 2L (CMT2L). METHODS: Ten positional and functional candidate genes were chosen among all known genes in this locus region by bioinformatics inqury. Mutation detection was performed by sequencing the exons and intron-exon junctions of the candidate genes. RESULTS: Eleven sequence variations, that included 5 heterozygous and 6 homozygous variations, were detected in the exons and flanking areas of the 10 candidate genes. All the variations showed no co-segregation with disease phenotype. CONCLUSION: Ten candidate genes(TAOK3, RAB35, RPLP0, PXN, RNF10, RHOF, VPS33A, RSN, DENR, RNP24) were ruled out as the disease-causing gene for CMT2L. Ten single nucleotide polymorphisms (SNP) were reported for the first time.

Base Sequence↗

Genetic variants in cyclooxygenase-2: Expression and risk of gastric cancer and its precursors in a Chinese population.

BACKGROUND & AIMS: To screen the genetic variants in cyclooxygenase-2 (COX-2), and evaluate their effects on COX-2 expression and risk of gastric cancer (GC) and its precursors, a population-based study was conducted in Linqu County, Shangdong Province, an area of China, a high-risk area of GC. METHODS: Genotypes were determined by polymerase chain reaction (PCR)-based denaturing high-performance liquid chromatography analysis in 248 GC cases and 1523 subjects with precancerous gastric lesions. COX-2 expression was detected by immunohistochemical analysis of biopsy specimens of 593 subjects selected at random from 1523 subjects. COX-2 transcriptional activity was examined by luciferase reporter gene assay. RESULTS: We found an increased risk of GC in subjects with -1195 AA genotype (adjusted odds ratio [OR], 2.33; 95% confidence interval [CI]: 1.45-3.75). Stratified analysis indicated that an elevated risk of GC was observed in subjects carrying the AA genotype and Helicobacter pylori infection (OR, 3.88; 95% CI: 1.46-10.34) or smoking (OR, 7.02; 95% CI: 2.19-22.48), and a high expression of COX-2 was found in subjects with AA genotype (OR, 1.84; 95% CI: 1.09-3.10) compared with GG genotype. The prevalence of COX-2 expression positivity varied markedly by histologic status, and the OR (OR, 5.35; 95% CI: 2.64-10.8) was significantly increased for dysplasia compared with superficial gastritis/chronic atrophic gastritis. Furthermore, tissue homogenate with H pylori infection could significantly stimulate COX-2 promoter activity driven by -1195A compared with the -1195G containing counterparts. CONCLUSIONS: These findings suggest that COX-2 polymorphisms may play an important role, at least in part, in developing GC in this high-risk population.

Adult↗

Genetic diversity among Mycobacterium avium complex strains recovered from children with and without human immunodeficiency virus infection.

The genetic diversity and molecular epidemiology of Mycobacterium avium complex (MAC) infections in children with and without human immunodeficiency virus (HIV) infection were evaluated. Isolates recovered from 136 children were subtyped by sequence analysis of a 360-bp region of the gene (hsp65) encoding a 65-kDa heat-shock protein. Twenty-one distinct hsp65 alleles were identified. On the basis of hsp65 genotype, 6 isolates were not MAC organisms. Of the remaining 130 samples, 61% were M. avium, 37% were Mycobacterium intracellulare, and 2% were species nonspecific MAC. Eighty-eight percent of the isolates obtained from HIV-infected children were M. avium. In contrast, only 38% of the isolates obtained from children without HIV infection were M. avium (chi2 test, P < .001). M. avium isolates were further subtyped by Southern blot analysis with insertion element IS1245. Taken together, no evidence for a single clonal M. avium strain causing infection was detected.

AIDS-Related Opportunistic Infections↗

Population systematics of chimpanzees using molar morphometrics.

When dental morphological variation within extant species is used as a guideline to partition variation within fossil samples into species, the underlying assumption is that fossil species are equivalent to extant species. This is the case despite the fact that dental morphology, which is commonly used to differentiate fossil species, is rarely used to differentiate extant species. Aspects of external morphology, ecology, behavior, breeding patterns, and molecular structure that are used to delineate living species are generally not available for fossils. In this paper, the utility of dental evidence for sorting fossil samples into species is evaluated by testing whether molar occlusal morphology is capable of sorting populations of Pan into the species and subspecies already well-established by nondental evidence. The dentitions of 341 chimpanzee individuals, sampled from regions throughout equatorial Africa, were sorted into 16 populations using rivers to demarcate the boundaries between populations. Digital-imaging software was used to measure 15 traits on the occlusal surface of each upper molar and 19 on each lower molar. After applying size adjustments, size-transformed and untransformed variables were subjected to discriminant analysis, with separate analyses carried out for each molar type. Results indicate that populations of Pan troglodytes and Pan paniscus are well differentiated at all molar positions. Populations of P. t. verus are distinct from other populations of P. troglodytes. Populations of P. t. troglodytes and P. t. schweinfurthii show close dental similarity. A distinct population is recognized at the Nigeria-Cameroon border, indicating the presence of P. t. vellerosus. The concordance between the patterns of diversity recognized by this study and other molecular and nonmolecular studies indicates that paleontological species that are similar to species of Pan in terms of size and patterns of diversification may be differentiated using molar morphology.

Africa↗

Microscopic electronic inhomogeneity in the high-Tc superconductor Bi2Sr2CaCu2O8+x.

The parent compounds of the copper oxide high-transition-temperature (high-Tc) superconductors are unusual insulators (so-called Mott insulators). Superconductivity arises when they are 'doped' away from stoichiometry. For the compound Bi2Sr2CaCu2O8+x, doping is achieved by adding extra oxygen atoms, which introduce positive charge carriers ('holes') into the CuO2 planes where the superconductivity is believed to originate. Aside from providing the charge carriers, the role of the oxygen dopants is not well understood, nor is it clear how the charge carriers are distributed on the planes. Many models of high-Tc superconductivity accordingly assume that the introduced carriers are distributed uniformly, leading to an electronically homogeneous system as in ordinary metals. Here we report the presence of an electronic inhomogeneity in Bi2Sr2CaCu2O8+x, on the basis of observations using scanning tunnelling microscopy and spectroscopy. The inhomogeneity is manifested as spatial variations in both the local density of states spectrum and the superconducting energy gap. These variations are correlated spatially and vary on the surprisingly short length scale of approximately 14 A. Our analysis suggests that this inhomogeneity is a consequence of proximity to a Mott insulator resulting in poor screening of the charge potentials associated with the oxygen ions left in the BiO plane after doping, and is indicative of the local nature of the superconducting state.

Journal Article↗

Perceived barriers in trying to eat healthier--results of a pan-EU consumer attitudinal survey.

Information on the perceived difficulties in trying to eat a healthier diet is important in assisting those in nutrition education devise more effective programmes. The objective of this study was to determine the main perceived barriers that people have in trying to eat a healthy diet in the 15 member states of the European Union (EU). A cross-sectional study in which quota-controlled nationally representative samples of approximately 1000 adults (15 years upwards) from each member state completed a face-to-face interview-assisted questionnaire. The most frequently mentioned perceived barriers to healthy eating concerned time and taste factors. Time-related factors were more important for younger respondents and those with a higher level of education, who appear to regard taste as being compromised by healthy eating. Variation exists both between member states and between demographic groups in the frequency of barriers mentioned. A lack of knowledge about healthy eating was not selected by many as an important barrier. A major obstacle to nutrition education is the fact that 70% of EU subjects believe their diets are already healthy. It may be that nutrition educators should concentrate on showing consumers how to evaluate their own diet appropriately in terms of fat, fibre, and fruit and vegetables. Food-based guidelines may be useful in this endeavour.

Adolescent↗