Background: Germline-speci®c differential DNA methylation that persists through fertilization and embryonic development is thought to be thè imprint' distinguishing the parental alleles of imprinted genes. If such methylation is to work as the imprinting mechanism, however, it has to be reprogrammed following each passage through the germline. Previous studies on maternally methylated genes have shown that their methylation imprints are ®rst erased in primordial germ cells (PGCs) and then re-established during oocyte growth.
A new imprinted gene has been discovered in mice using the technique of restriction landmark genomic scanning (RLGS) with methylation sensitive enzymes. Eight out of 3,100 strain-specific NotI and BssHII spots were identified as imprinted in reciprocal F1 hybrids. Subsequently, we isolated a genomic clone for one locus on proximal chromosome 11 near the Glns locus, an imprinted region in uniparental disomic mice, and its corresponding cDNA clone. Expression of this transcript from the paternal allele was established using RT-PCR of reciprocal F1-hybrid mice. The amino-acid sequence deduced from the cDNA showed significant homology to the U2 small nuclear ribonucleoprotein auxiliary factor 35 kDa subunit.
The purpose of this study was to determine the viability of Tongue Coating Index, which is a new method for evaluating tongue-coating status. To determine the reliability and reproducibility of our new evaluation criteria (Score 0: Tongue coating not visible; Score 1: Tongue coating thin, papillae of tongue visible; Score 2: Tongue coating very thick, papillae of tongue not visible), 10 observers evaluated 20 photographs of tongues. Each tongue surface was divided into nine sections. Observers evaluated each section according to our new criteria and each score for tongue-coating status was recorded in the pertinent section of the Tongue Coating Record form. They repeated the same evaluation 2 weeks after the first evaluation. The relationship between the scores obtained and number of oral microorganisms was investigated in 50 edentulous patients. Tongue coating was collected from the tongue surface after evaluation of tongue-coating status. The total number of anaerobic bacteria and the number of Candida species were counted from the specimens collected. Interobserver agreement and intraobserver agreement were 0.66 and 0.80 by Cohen's kappa, respectively. No significant difference was observed in the number of Candida species among the three scores. The number of total anaerobic bacteria, however, was significantly different among the scores (P < 0.05). Therefore, we conclude that our method for evaluating tongue-coating status offers new criteria that are superior in reliability and reproducibility, and that also reflect the total number of anaerobic bacteria present on the dorsum of the tongue.
Maintenance-type DNA methyltransferase (Dnmt1) is usually down-regulated in non-proliferating cells. In the present study, we detected significant expression of Dnmt1 protein in adult mouse brain where the majority of the cells are in a post-mitotic state. A significant amount of Dnmt1 protein was fractionated into the post-nuclear fraction for both cerebrum and cerebellum. The Dnmt1 in this fraction was enzymatically active. An immunofluorescence study revealed that Dnmt1 protein was mainly expressed in neurons and seemed to be localized in the cytoplasmic compartment. Primary culturing of neurons confirmed the expression and localization of Dnmt1 in the cytoplasmic compartment. The findings that the Dnmt1 transcript in the brain utilized the somatic-type exon and that the apparent size of the Dnmt1 protein in the cytoplasm was identical to that in proliferating culture cells indicate that the cytoplasmic Dnmt1 in neurons was of the somatic-type.
Oral hypofunction, resulting from a combined decrease in multiple oral functions, may affect systemic-condition deterioration; however, few studies have examined the association between oral hypofunction and general health among older adults. In this cross-sectional study, we examined the relationship between oral hypofunction and sarcopenia in community-dwelling older adults. We included 878 adults (268 men and 610 women, mean age 76.5 ± 8.3 years). Tongue coating index, oral moisture, occlusal force, oral diadochokinesis (/pa/,/ta/,/ka/), tongue pressure, mas-ticatory function, and swallowing function were evaluated as indicators of oral hypofunction. Grip strength, gait speed, and skeletal muscle mass index were measured as diagnostic sarcopenia parameters. The association between oral hypofunction and sarcopenia was examined via logistic regression using sarcopenia as the dependent variable. Oral hypofunction prevalence was 50.5% overall, 40.3% in men, and 54.9% in women. The prevalence of sarcopenia was 18.6% overall, 9.7% in men, and 22.5% in women. A logistic regression showed oral hypofunction, age, body mass index, higher-level functional capacity, and serum albumin level were significantly associated with sarcopenia. Sarcopenia occurred at an increased frequency in patients diagnosed with oral hypofunction (odds ratio: 1.59, 95% confidence interval: 1.02–2.47); accordingly, oral hypofunction appears to be significantly associated with sarcopenia.
Our results confirm that masticatory function was associated with the progression to pre-frailty or frailty among community-dwelling individuals 65 years and older over the 2-year period of this longitudinal study. Of the masticatory function items evaluated, mixing ability and subjective chewing ability were associated with frailty progression.
A systematic screen termed the allelic message display (AMD) was developed for the hunting of imprinted genes. In AMD, differential display PCR is adopted to image allelic expression status of multiple polymorphic transcripts in two parental mouse strains, reciprocal F 1 hybrids and pooled backcross progenies. From the displayed patterns, paternally and maternally expressed transcripts can be unequivocally identified. The effectiveness of AMD screening was clearly demonstrated by the identification of a paternally expressed gene Impact on mouse chromosome 18, the predicted product of which belongs to the YCR59c͞yigZ hypothetical protein family composed of yeast and bacterial proteins with currently unknown function. In contrast with previous screening methods necessitating positional cloning efforts or generation of parthenogenetic embryos, this approach requires nothing particular but appropriately crossed mice and can be readily applied to any tissues at various developmental stages. Hence, AMD would considerably accelerate the identification of imprinted genes playing pivotal roles in mammalian development and the pathogenesis of various diseases.
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