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1.
Oxf Med Case Reports ; 2024(3): omae009, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38532755

ABSTRACT

An 80-year-old man presented in December with the main complaint of jaundice. Blood tests revealed hemolytic anemia and renal dysfunction. Positive syphilis serology results led to a diagnosis of untreated latent syphilis. A positive direct Coombs test led to a diagnosis of autoimmune hemolytic anemia (AIHA). Antibiotics were started for the syphilis, with improvement in the anemia and renal dysfunction observed. However, paroxysmal intravascular hemolysis occurred after his discharge. Based on a positive Donath-Landsteiner (D-L) test, paroxysmal cold hemoglobinuria (PCH) diagnosis was made. The hemolytic anemia improved after further treatment for syphilis, and further avoiding exposure to cold.

2.
Arch Oral Biol ; 154: 105761, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37421827

ABSTRACT

OBJECTIVE: Childhood is an important period for lip-closing strength (LCS) development, and failure to acquire LCS during childhood leads to various adverse health effects, such as mouth breathing. The purpose of this study was to examine the effectiveness of device-free lip and facial training in preschool children. DESIGN: The participants were divided into training and control groups. Both groups comprised 123 children aged 3-4 years, and only the training group received lip and facial training (i.e., opening and closing the lips and protruding the tongue) for 1 year. A two-way repeated measures analysis of variance was applied to compare the interaction effects of LCS and facial linear distance and angle by year (initial year vs. 1 year later) and group (training vs. control group). In addition, paired t-tests were used to test the changes in LCS and facial linear distance and angle after 1 year in both groups. Furthermore, the same analysis was performed in children with weak LCS in both groups (incompetent lip seal [ILS]). RESULTS: The LCS of children in the training group significantly increased after training compared with that in the control group, whether the analysis included all children or children with ILS alone. Lip and facial training for children with ILS reduced both the upper and lower lip protrusion; children with ILS without training had increased lip protrusion after 1 year. CONCLUSIONS: Lip and facial training for children with ILS effectively improved LCS and lip morphology, thereby preventing increased lip protrusion.


Subject(s)
Face , Lip , Child, Preschool , Humans , Lip/anatomy & histology , Face/anatomy & histology , Tongue , Cephalometry
3.
J Clin Med ; 11(20)2022 Oct 15.
Article in English | MEDLINE | ID: mdl-36294410

ABSTRACT

Tissue-specific stem cells exist in tissues and organs, such as skin and bone marrow. However, their pluripotency is limited compared to embryonic stem cells. Culturing primary cells on plastic tissue culture dishes can result in the loss of multipotency, because of the inability of tissue-specific stem cells to survive in feeder-less dishes. Recent findings suggest that culturing primary cells in medium containing feeder cells, particularly genetically modified feeder cells expressing growth factors, may be beneficial for their survival and proliferation. Therefore, the aim of this study was to elucidate the role of genetically modified human feeder cells expressing growth factors in maintaining the integrity of primary cultured human deciduous dental pulp cells. Feeder cells expressing leukemia inhibitory factor, bone morphogenetic protein 4, and basic fibroblast growth factor were successfully engineered, as evidenced by PCR. Co-culturing with mitomycin-C-treated feeder cells enhanced the proliferation of newly isolated human deciduous dental pulp cells, promoted their differentiation into adipocytes and neurons, and maintained their stemness properties. Our findings suggest that genetically modified human feeder cells may be used to maintain the integrity of primary cultured human deciduous dental pulp cells.

4.
Clin Exp Dent Res ; 8(1): 209-216, 2022 02.
Article in English | MEDLINE | ID: mdl-34499413

ABSTRACT

OBJECTIVES: Weakening of lip-closing strength (LCS) associated with an incompetent lip seal (ILS) may affect the oral balance between the lip and tongue pressures. The purpose of this study was to evaluate the effects of lip-closing training in children with lower LCS and/or abnormal habits across different age groups and to compare its effects on increasing LCS in children with malocclusion and/or oral habits. MATERIAL AND METHODS: Lip-closing training was performed by 154 Japanese children aged 3-12 years using a specialized training device at home for 3 months. Children with oral habits and/or exhibiting less than standard LCS were included. LCS was measured using a digital strain force gauge at a dental clinic at the beginning (T0) and after each month (after 3 months: T3). RESULTS: Children had higher LCS responses after lip-closing training. The first month of lip-closing training was more effective than the subsequent months. With lip-closing training, the LCS increased from an average of 6.2 N (T0) to 11.4 N (T3) in Group I, 7.9 N (T0) to 12.8 N (T3) in Group II, and 6.8 N to 11.4 N in Group III. Anterior cross bite, including reverse bite, open bite, and tongue thrusting, significantly reduced training effects. CONCLUSION: Our findings showed that lower LCS in children with ILS resulted in greater responses to lip-closing training in a short period, but oral dysfunction, such as abnormal habits, inhibited the positive effects of training. Our results suggest that less detrimental effects of malocclusion and abnormal oral habits lip-closing training enhances LCS in younger children.


Subject(s)
Facial Muscles , Malocclusion , Child , Facial Muscles/physiology , Humans , Lip/physiology , Malocclusion/therapy , Pressure , Tongue
5.
Sleep Biol Rhythms ; 20(2): 247-254, 2022 Apr.
Article in English | MEDLINE | ID: mdl-38469249

ABSTRACT

Bright light is a primary zeitgeber (synchronizer) for the central circadian pacemaker in humans. Recently, head-mounted devices for light therapy have been developed to treat patients suffering from circadian rhythm sleep disorders. In this study, to evaluate the influence of the light incident angle of head-mounted devices on the human circadian pacemaker, we examined the effects of bright light (ca.10000 lx) from two different angles (55° vs. 28°) on the suppression of melatonin secretion at night. Twenty-nine subjects (25.1 ± 6.3 SD years) participated in the present study. The subjects were kept under dim light conditions (< 5 lx) from 4 h before their habitual bedtime, followed by exposure to 1 h of bright light at two different angles during their habitual bedtime. Saliva samples were collected every hour under dim light conditions and then collected every 30 min during the bright light exposure. To assess the effect of the light incident angle on ipRGCs mediating light-evoked pupillary constriction, pupil size was measured in before and after exposure to bright light. Melatonin suppression in the group exposed to light at 28° was significantly higher than that in the group with light at 55° (p < 0.001). The pupillary constriction was significantly greater in the group exposed to light at 28° than that in the group with light at 55° (p < 0.001). The present findings suggest that the light incident angle is an important factor for bright light therapy and should be considered to effectively use head-mounted devices in home and clinical settings.

6.
Cells ; 10(12)2021 11 28.
Article in English | MEDLINE | ID: mdl-34943845

ABSTRACT

Alkaline phosphatase (ALP) is a ubiquitous membrane-bound glycoprotein capable of providing inorganic phosphate by catalyzing the hydrolysis of organic phosphate esters, or removing inorganic pyrophosphate that inhibits calcification. In humans, four forms of ALP cDNA have been cloned, among which tissue-nonspecific ALP (TNSALP) (TNSALP) is widely distributed in the liver, bone, and kidney, making it an important marker in clinical and basic research. Interestingly, TNSALP is highly expressed in juvenile cells, such as pluripotent stem cells (i.e., embryonic stem cells and induced pluripotent stem cells (iPSCs)) and somatic stem cells (i.e., neuronal stem cells and bone marrow mesenchymal stem cells). Hypophosphatasia is a genetic disorder causing defects in bone and tooth development as well as neurogenesis. Mutations in the gene coding for TNSALP are thought to be responsible for the abnormalities, suggesting the essential role of TNSALP in these events. Moreover, a reverse-genetics-based study using mice revealed that TNSALP is important in bone and tooth development as well as neurogenesis. However, little is known about the role of TNSALP in the maintenance and differentiation of juvenile cells. Recently, it was reported that cells enriched with TNSALP are more easily reprogrammed into iPSCs than those with less TNSALP. Furthermore, in bone marrow stem cells, ALP could function as a "signal regulator" deciding the fate of these cells. In this review, we summarize the properties of ALP and the background of ALP gene analysis and its manipulation, with a special focus on the potential role of TNSALP in the generation (and possibly maintenance) of juvenile cells.


Subject(s)
Alkaline Phosphatase/metabolism , Cell Differentiation , Animals , Humans , Isoenzymes/metabolism , Models, Biological , Research , Signal Transduction
7.
Biol Proced Online ; 23(1): 12, 2021 Jun 11.
Article in English | MEDLINE | ID: mdl-34116635

ABSTRACT

BACKGROUND: Expression of stemness factors, such as octamer-binding transcription factor 3/4 (OCT3/4), sex determining region Y-box 2 (SOX2), and alkaline phosphatase (ALP) in human deciduous tooth-derived dental pulp cells (HDDPCs) can be assessed through fixation and subsequent immuno- or cytochemical staining. Fluorescence-activated cell sorting (FACS), a powerful system to collect cells of interest, is limited by the instrument cost and difficulty in handling. Magnetic-activated cell sorting is inexpensive compared to FACS, but is confined to cells with surface expression of the target molecule. In this study, a simple and inexpensive method was developed for the molecular analysis of immuno- or cytochemically stained cells with intracellular expression of a target molecule, through isolation of a few cells under a dissecting microscope using a mouthpiece-controlled micropipette. RESULTS: Two or more colored cells (~ 10), after staining with a chromogen such a 3,3'-diaminobenzidine, were successfully segregated from unstained cells. Expression of glyceraldehyde 3-phosphate dehydrogenase, a housekeeping gene, was discernible in all samples, while the expression of stemness genes (such as OCT3/4, SOX2, and ALP) was confined to positively stained cells. CONCLUSION: These findings indicate the fidelity of these approaches in profiling cells exhibiting cytoplasmic or nuclear localization of stemness-specific gene products at a small-scale.

8.
Environ Health Prev Med ; 26(1): 11, 2021 Jan 21.
Article in English | MEDLINE | ID: mdl-33478389

ABSTRACT

BACKGROUND: Systemic and local factors may lead to disruption of craniofacial growth and development, causing an imbalance between the orofacial skeleton, muscle and soft tissue, dental occlusion, and the dental arch during growth periods. We aimed to reveal whether the prevalence of incompetent lip seal (ILS) varies with age and region, as well as to clarify the factors related to an ILS, in a national, large-scale epidemiological study. METHODS: We surveyed 3399 children, from 3 to 12 years of age, visiting 66 pediatric dental clinics throughout Japan. For this survey, we employed a questionnaire consisting of 44 questions regarding daily health conditions and lifestyle habits. We evaluated the differences in ILS prevalence by age and region (using a Cochran-Armitage test for trend and a Kruskal-Wallis test), and the relationship between ILS and factors investigated in the questionnaire (using Spearman's rank correlation coefficient). RESULTS: We observed that 30.7% of Japanese children exhibited an ILS and that the ILS rate increased with age (p < 0.001). There were no regional differences in the rate of ILS in Japanese children (p = 0.506). We revealed that 12 of 44 survey items exhibited a statistically significant correlation with ILS (p < 0.001), using Spearman's rank correlation coefficient. These items involved orofacial morphology, mouth breathing, and possibly, allergic rhinitis. CONCLUSION: The rate of ILS seems to increase with age in children, throughout Japan. Therefore, this disorder may not self-correct during the growth periods in these children. Guidelines are required for pediatric dentists to recognize ILS among children aged 3-12 years.


Subject(s)
Lip/abnormalities , Child , Child, Preschool , Cross-Sectional Studies , Female , Humans , Japan/epidemiology , Male , Prevalence
9.
Cranio ; 39(5): 405-411, 2021 Sep.
Article in English | MEDLINE | ID: mdl-31469617

ABSTRACT

Objective: To examine whether incompetent lip seal (ILS) influences the form of facial soft tissue.Methods: Four hundred forty-four preschool children 3-5 years of age were selected. The images of the subjects' facial surface were obtained with a three-dimensional laser scanner. Coordinates of 16 facial landmarks were established and identified on the three-dimensional facial images, and the differences between children with (wILS) and without ILS (woILS) were measured.Results: The angle of sagittal facial convexity, excluding the nose, in 4- and 5-year-old children was significantly smaller in wILS children than in woILS children. The nasal prominence angle and the protrusion angle of lips in wILS children were significantly smaller than those in woILS children, at all ages.Conclusion: Children with ILS have anteriorly prominent subnasales and lips and flatter noses. The influence of ILS on facial form begins to appear even before 3 years of age.


Subject(s)
Face , Lip , Cephalometry , Child, Preschool , Face/anatomy & histology , Humans , Imaging, Three-Dimensional
10.
J Clin Med ; 9(9)2020 Sep 02.
Article in English | MEDLINE | ID: mdl-32887316

ABSTRACT

Pluripotent stem cells are classified as naïve and primed cells, based on their in vitro growth characteristics and potential to differentiate into various types of cells. Human-induced pluripotent stem cells (iPSCs, also known as epiblast stem cells [EpiSCs]) have limited capacity to differentiate and are slightly more differentiated than naïve stem cells (NSCs). Although there are several in vitro protocols that allow iPSCs to differentiate into pancreatic lineage, data concerning generation of ß-cells from these iPSCs are limited. Based on the pluripotentiality of NSCs, it was hypothesized that NSCs can differentiate into pancreatic ß-cells when placed under an appropriate differentiation induction condition. We examined whether NSCs can be efficiently induced to form potentially pancreatic ß cells after being subjected to an in vitro protocol. Several colonies resembling in vitro-produced ß-cell foci, with ß-cell-specific marker expression, were observed when NSC-derived embryoid bodies (EBs) were induced to differentiate into ß-cell lineage. Conversely, EpiSC-derived EBs failed to form such foci in vitro. Intrapancreatic grafting of the in vitro-formed ß-cell foci into nude mice (BALB/c-nu/nu) generated a cell mass containing insulin-producing cells (IPCs), without noticeable tumorigenesis. These NSCs can be used as a promising resource for curing type 1 diabetes.

11.
Int J Mol Sci ; 20(19)2019 Oct 03.
Article in English | MEDLINE | ID: mdl-31623314

ABSTRACT

We aimed to immortalize primarily isolated human deciduous tooth-derived dental pulp cells (HDDPCs) by transfection with piggyBac (PB)-based transposon vectors carrying E7 from human papilloma virus 16 or complementary DNA (cDNA) encoding human telomerase reverse transcriptase (hTERT). HDDPCs were co-transfected with pTrans (conferring PB transposase expression) + pT-pac (conferring puromycin acetyltransferase expression) + pT-tdTomato (conferring tdTomato cDNA expression) and pT-E7 (conferring E7 expression) or pTrans + pT-pac + pT-EGFP (conferring enhanced green fluorescent protein cDNA expression) + pT-hTERT (conferring hTERT expression). After six days, these cells were selected in medium containing 5 µg/mL puromycin for one day, and then cultured in normal medium allowing cell survival. All resultant colonies were harvested and propagated as a pool. Stemness and tumorigenic properties of the established cell lines ("MT_E7" for E7 and "MT_hTERT" for hTERT) with untransfected parental cells (MT) were examined. Both lines exhibited proliferation similar to that of MT, with alkaline phosphatase activity and stemness-specific factor expression. They displayed differentiation potential into multi-lineage cells with no tumorigenic property. Overall, we successfully obtained HDDPC-derived immortalized cell lines using a PB-based transfection system. The resultant and parental cells were indistinguishable. Thus, E7 and hTERT could immortalize HDDPCs without causing cancer-associated changes or altering phenotypic properties.


Subject(s)
Cell Differentiation , DNA Transposable Elements , Dental Pulp/cytology , Stem Cells/cytology , Stem Cells/metabolism , Cell Differentiation/genetics , Cell Line, Transformed , Cell Transformation, Neoplastic , Female , Genetic Vectors/genetics , Humans , Oncogene Proteins, Viral/genetics , Stem Cells/pathology , Telomerase/genetics , Telomerase/metabolism , Tooth, Deciduous , Transfection
12.
Int J Mol Sci ; 20(7)2019 Apr 03.
Article in English | MEDLINE | ID: mdl-30987116

ABSTRACT

Stage-specific embryonic antigen 1 (SSEA-1) is an antigenic epitope (also called CD15 antigen) defined as a Lewis X carbohydrate structure and known to be expressed in murine embryonal carcinoma cells, mouse embryonic stem cells (ESCs), and murine and human germ cells, but not human ESCs/induced pluripotent stem cells (iPSCs). It is produced by α1,3-fucosyltransferase IX gene (FUT9), and F9 ECCs having a disrupted FUT9 locus by gene targeting are reported to exhibit loss of SSEA-1 expression on their cell surface. Mouse ESCs are pluripotent cells and therefore known as "naïve stem cells (NSCs)." In contrast, human ESCs/iPSCs are thought to be epiblast stem cells (EpiSCs) that are slightly more differentiated than NSCs. Recently, it has been demonstrated that treatment of EpiSCs with several reprograming-related drugs can convert EpiSCs to cells similar to NSCs, which led us to speculate that SSEA-1 may have been expressed in these NSC-like EpiSCs. Immunocytochemical staining of these cells with anti-SSEA-1 revealed increased expression of this epitope. RT-PCR analysis also confirmed increased expression of FUT9 transcripts as well as other stemness-related transcripts such as REX-1 (ZFP42). These results suggest that SSEA-1 can be an excellent marker for human NSCs.


Subject(s)
Cell Membrane/metabolism , Dental Pulp/cytology , Induced Pluripotent Stem Cells/cytology , Lewis X Antigen/metabolism , Tooth, Deciduous/cytology , Animals , Colony-Forming Units Assay , Humans , Induced Pluripotent Stem Cells/metabolism , Mice, Inbred BALB C , Mice, Nude
13.
Sci Rep ; 9(1): 1490, 2019 02 06.
Article in English | MEDLINE | ID: mdl-30728386

ABSTRACT

Human tissue-specific stem cells (hTSCs), found throughout the body, can differentiate into several lineages under appropriate conditions in vitro and in vivo. By transfecting terminally differentiated cells with reprogramming factors, we previously produced induced TSCs from the pancreas and hepatocytes that exhibit additional properties than iPSCs, as exemplified by very low tumour formation after xenogenic transplantation. We hypothesised that hTSCs, being partially reprogrammed in a state just prior to iPSC transition, could be isolated from any terminally differentiated cell type through transient reprogramming factor overexpression. Cytochemical staining of human deciduous tooth-derived dental pulp cells (HDDPCs) and human skin-derived fibroblasts following transfection with Yamanaka's factors demonstrated increased ALP activity, a stem cell marker, three weeks after transfection albeit in a small percentage of clones. Repeated transfections (≤3) led to more efficient iPSC generation, with HDDPCs exhibiting greater multipotentiality at two weeks post-transfection than the parental intact HDDPCs. These results indicated the utility of iPSC technology to isolate TSCs from HDDPCs and fibroblasts. Generally, a step-wise loss of pluripotential phenotypes in ESCs/iPSCs occurs during their differentiation process. Our present findings suggest that the reverse phenomenon can also occur upon repeated introduction of reprogramming factors into differentiated cells such as HDDPCs and fibroblasts.


Subject(s)
Cell Culture Techniques/methods , Dental Pulp/pathology , Induced Pluripotent Stem Cells/cytology , Cell Differentiation/physiology , Cells, Cultured , Cellular Reprogramming/physiology , Fibroblasts/cytology , Humans , Multipotent Stem Cells/cytology , Skin/cytology , Tooth, Deciduous/cytology
14.
Arch Oral Biol ; 92: 57-61, 2018 Aug.
Article in English | MEDLINE | ID: mdl-29753207

ABSTRACT

OBJECTIVE: Mouth breathing syndrome (MBS) is defined as a set of signs and symptoms that may be completely or incompletely present in subjects who, for various reasons, replace the correct pattern of nasal breathing with an oral or mixed pattern. It is important to identify the relevant factors affecting MBS in order to diagnose its cause since breathing obstructions can result from multiple factors. The purpose of this study is to clarify the relevant factors and the interrelationships between factors affecting MBS among children. DESIGN: We surveyed 380 elementary school children from 6 to 12 years in age. The questionnaire consisted of 44 questions regarding their daily health conditions and lifestyle habits and was completed by the children's guardians. A factor analysis was performed to classify closely related questions into their respective factors and to examine the strength of the correlation between the newly revealed factors. RESULTS: Twenty-six out of the 44 questions were selected, and they were classified into seven factors. Factors 1-7 were defined as "Incompetent lip seal", "Diseases of the nose and throat", "Eating and drinking habits", "Bad breath", "Problems with swallowing and chewing", "Condition of teeth and gums", and "Dry lips", respectively. There were also correlations between these factors themselves. CONCLUSION: MBS was categorized according to 7 major factors. Because Factor 1 was defined as "Incompetent lip seal", which was representative of the physical appearance of mouth breathers and correlated with other factors, we suggested that MBS should consist of 7 factors in total.


Subject(s)
Mouth Breathing/etiology , Child , Factor Analysis, Statistical , Female , Humans , Japan , Male , Mouth Breathing/physiopathology , Risk Factors , Surveys and Questionnaires
15.
Int J Mol Sci ; 18(8)2017 Aug 02.
Article in English | MEDLINE | ID: mdl-28767080

ABSTRACT

In vivo inoculation of cells such as tumor cells and induced pluripotent stem (iPS)/embryonic stem (ES) cells into immunocompromised mice has been considered as a powerful technique to evaluate their potential to proliferate or differentiate into various cell types originating from three germ cell layers. Subcutaneous grafting and grafting under the kidney capsule have been widely used for this purpose, but there are some demerits such as the requirement of a large number of tumor cells for inoculation and frequent failure of tumorigenesis. Therefore, grafting into other sites has been explored, including intratesticular or intramuscular grafting as well as grafting into the cochleae, liver, or salivary glands. In this study, we found that intrapancreatic parenchymal injection of cells is useful for allowing a small number of cells (~15 × 10³ cells or ~30 cell clumps µL-1·site-1) to proliferate and sometimes differentiate into various types of cells. It requires only surgical exposure of the pancreas over the dorsal skin and subsequent injection of cells towards the pancreatic parenchyma under dissecting microscope-based observation using a mouthpiece-controlled glass micropipette. We now name this technology "intrapancreatic parenchymal cell transplantation (IPPCT)", which will be useful, especially when only a small number of cells or colonies are available.


Subject(s)
Cell Proliferation , Human Embryonic Stem Cells , Induced Pluripotent Stem Cells , Liver/metabolism , Animals , Cell Line , Female , Heterografts , Human Embryonic Stem Cells/metabolism , Human Embryonic Stem Cells/transplantation , Humans , Induced Pluripotent Stem Cells/metabolism , Induced Pluripotent Stem Cells/transplantation , Mice , Mice, Inbred BALB C , Mice, Nude
16.
J Investig Clin Dent ; 8(4)2017 Nov.
Article in English | MEDLINE | ID: mdl-27641728

ABSTRACT

AIM: The aim of the present study was to prove that primary cells enriched with stem cells are more easily reprogrammed to generate induced pluripotent stem (iPS) cells than those with scarce numbers of stem cells. METHODS: We surveyed the alkaline phosphatase (ALP) activity in five primarily-isolated human deciduous teeth-derived dental pulp cells (HDDPC) with cytochemical staining to examine the possible presence of stem cells. Next, the expression of stemness-specific factors, such as OCT(Octumer-binding transcription factor)3/4, NANOG, SOX2(SRY (sex determining region Y)-box 2), CD90, muscle segment homeodomain homeobox (MSX) 1, and MSX2, was assessed with a reverse transcription polymerase chain reaction method. Finally, these isolated HDDPC were transfected with plasmids carrying genes coding Yamanaka factors to determine whether these cells could be reprogrammed to generate iPS cells. RESULTS: Of the five primarily-isolated HDDPC, two (HDDPC-1 and -5) exhibited higher degrees of ALP activity. OCT-3/4 expression was also prominent in those two lines. Furthermore, these two lines proliferated faster than the other three lines. The transfection of HDDPC with Yamanaka factors resulted in the generation of iPS cells from HDDPC-1 and -5. CONCLUSION: The number of cells with the stemness property of HDDPC differs among individuals, which suggests that HDDPC showing an increased expression of both ALP and OCT-3/4 can be more easily reprogrammed to generate iPS cells after the forced expression of reprogramming factors.


Subject(s)
Alkaline Phosphatase/metabolism , Dental Pulp/cytology , Dental Pulp/metabolism , Induced Pluripotent Stem Cells , Octamer Transcription Factor-3/biosynthesis , Humans , Tooth, Deciduous
17.
World J Hepatol ; 8(5): 282-90, 2016 Feb 18.
Article in English | MEDLINE | ID: mdl-26925202

ABSTRACT

AIM: To address the effect of heat-shock protein 90 (HSP90) inhibitors on the release of the hepatitis C virus (HCV), a cell culture-derived HCV (JFH1/HCVcc) from Huh-7 cells was examined. METHODS: We quantified both the intracellular and extracellular (culture medium) levels of the components (RNA and core) of JFH-1/HCVcc. The intracellular HCV RNA and core levels were determined after the JFH1/HCVcc-infected Huh-7 cells were treated with radicicol for 36 h. The extracellular HCV RNA and core protein levels were determined from the medium of the last 24 h of radicicol treatment. To determine the possible role of the HSP90 inhibitor in HCV release, we examined the effect of a combined application of low doses of the HSP90 inhibitor radicicol and the RNA replication inhibitors cyclosporin A (CsA) or interferon. Finally, we statistically examined the combined effect of radicicol and CsA using the combination index (CI) and graphical representation proposed by Chou and Talalay. RESULTS: We found that the HSP90 inhibitors had greater inhibitory effects on the HCV RNA and core protein levels measured in the medium than inside the cells. This inhibitory effect was observed in the presence of a low level of a known RNA replication inhibitor (CsA or interferon-α). Treating the cells with a combination of radicicol and cyclosporin A for 24 h resulted in significant synergy (CI < 1) that affected the release of both the viral RNA and the core protein. CONCLUSION: In addition to having an inhibitory effect on RNA replication, HSP90 inhibitors may interfere with an HCV replication step that occurs after the synthesis of viral RNA, such as assembly and release.

18.
Cell Med ; 8(1-2): 9-23, 2015 Dec 17.
Article in English | MEDLINE | ID: mdl-26858904

ABSTRACT

Feeder cells are generally required to maintain embryonic stem cells (ESCs)/induced pluripotent stem cells (iPSCs). Mouse embryonic fibroblasts (MEFs) isolated from fetuses and STO mouse stromal cell line are the most widely used feeder cells. The aim of this study was to determine which cells are suitable for establishing iPSCs from human deciduous tooth dental pulp cells (HDDPCs). Primary cultures of HDDPCs were cotransfected with three plasmids containing human OCT3/4, SOX2/KLF4, or LMYC/LIN28 and pmaxGFP by using a novel electroporation method, and then cultured in an ESC qualified medium for 15 days. Emerging colonies were reseeded onto mitomycin C-treated MEFs or STO cells. The colonies were serially passaged for up to 26 passages. During this period, colony morphology was assessed to determine whether cells exhibited ESC-like morphology and alkaline phosphatase activity to evaluate the state of cellular reprogramming. HDDPCs maintained on MEFs were successfully reprogrammed into iPSCs, whereas those maintained on STO cells were not. Once established, the iPSCs were maintained on STO cells without loss of pluripotency. Our results indicate that MEFs are better feeder cells than STO cells for establishing iPSCs. Feeder choice is a key factor enabling efficient generation of iPSCs.

19.
Arch Oral Biol ; 59(12): 1391-9, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25214309

ABSTRACT

OBJECTIVE: To evaluate morphological differences of the facial soft tissue surface between male Japanese adults and children. DESIGN: 20 adult Japanese males (average age 28 years) and 20 Japanese boys (average age 5.5 years) with normal occlusion were selected for this study. The images of the subjects' facial surface were obtained with a 3-D laser scanner. To evaluate the three-dimensional morphological differences of the facial soft tissue, we transformed the coordinates of 16 facial landmarks to a new reference plane and compared the adults' and children's facial form drawn to the same scale in the same coordinate system. RESULTS: The morphological difference ratio of the lower facial area was higher than in the upper facial area, and the nose and lower face changed more forward than downward. The morphological difference ratio of the mid face width was smaller than other areas. CONCLUSION: Our study suggests that the morphological facial soft tissue differences between Japanese adults and children are more forward and downward than laterally, manifesting in a facial form of adults that is deeper and narrow.


Subject(s)
Face/anatomy & histology , Imaging, Three-Dimensional/instrumentation , Lasers , Maxillofacial Development , Adult , Child, Preschool , Cross-Sectional Studies , Humans , Japan , Male
20.
Intern Med ; 53(7): 805-8, 2014.
Article in English | MEDLINE | ID: mdl-24694502

ABSTRACT

A 48-year-old man visited our hospital complaining of a tender mass in the left side of the neck. He was diagnosed with tuberculous lymphadenitis based on the results of a biopsy. Shortly after the diagnosis, oral aphthae, erythema nodosum-like lesions on the lower legs and genital ulcers developed. A diagnosis of cutaneous tuberculosis was ruled out according to a negative mycobacterial culture of tissues obtained from stained smears and lesional biopsy specimens. The patient's symptoms remitted following the introduction of antituberculous therapy. We assume that tuberculous lymphadenitis was strongly associated with the appearance of Behçet's disease-like symptoms in this case.


Subject(s)
Behcet Syndrome/etiology , Lymph Nodes/pathology , Tuberculosis, Lymph Node/complications , Behcet Syndrome/diagnosis , Biopsy , Diagnosis, Differential , Humans , Lymph Nodes/microbiology , Male , Middle Aged , Mycobacterium tuberculosis/isolation & purification , Tuberculosis, Lymph Node/diagnosis
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