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Sunil Richardson,James S. Hoyt,Rohit K. Khosla,Rakshit Vijay Sinai Khandeparker,Vihang Y. Sukhadia,Nisheet Agni 대한구강악안면외과학회 2016 대한구강악안면외과학회지 Vol.42 No.2
Objectives: To evaluate the effectiveness of regenerative tissue matrix (Alloderm) as an oral layer for difficult anterior palatal fistula closure. Materials and Methods: The authors have tested the feasibility of a novel surgical technique of adding a regenerative tissue matrix (Alloderm) as an oral layer for closure of recalcitrant large anterior palatal fistulae and report the outcome of the first 12 patients in this pilot study. Patients with recurrent large fistula who otherwise would require either a local pedicled flap, free flap, or an obturator were treated with this technique and followed up for at least 6 months to monitor the progress of healing. Results: Of the 12 patients, 8 patients (66.7%) had complete closure of the fistula, and 2 patients (16.7%) showed reduction in size of the fistula to the extent that symptoms were eliminated, for an overall success rate of 83.3% (10/12 patients). Premature graft loss and recurrence of the fistula were noted in 2 patients (16.7%). Conclusion: Alloderm provided an adequate barrier allowing healing to occur unimpeded and allowed closure of the palatal fistula. In our experience, this new technique using regenerative tissue matrix as an adjunct to the oral layer in large anterior palatal fistula has an advantage compared to other more invasive complex procedures and has been shown to provide satisfactory results.
Characterization of Ion Transporter Proteins Expressed in Rat Parotid Acinar Cell Line, Par-C10
Park, Keerang,Chang, Heesoon,Richardson, Linda A.,Lee, Young-Ill,Choi, Jiun,Melvin, James E. Korean Academy of Oral Biology and the UCLA Dental 2001 International Journal of Oral Biology Vol.26 No.1
In this study we characterized the ion transporter proteins present in the immortalized rat parotid acinar cell line, Par-C10 to determine whether it is an appropriate in vitro model for studies of fluid and electrolyte secretion in rat parotid acinar cells. Western blot analyses for CIC-2 chloride channel and Na^+/H^+ exchanger isoform 1 and 2 (NHE1 and NHE2) showed that CIC-2 and NHE1 proteins express in Par-C10, whereas NHE2 protein expression was not detectable. These observations in Par-C10 cell line are consistent-Polymerase Chain Reaction (RT-PCR) of Par-C10 or rat parotid acinar cell cDNA for CIC-2 and CIC-3 chloride channels demonstrated that both cell types express the transcripts for these chloride channels. To determine whether Par-C10 cell line is derived from acinar or ductal cells we performed semi-quantitative RT-PCR for four different members of Na^+/H^+ exchanger gene family (NHE1, NHE2, NHE3, and NHE4) including control amplification for acinar-specific amylase or ductal-specific kallikrein. Par-C10 cells express transcripts for amylase abundantly, but not kallikrein. Our semi-quantitative RT-PCR results also showed that, like native parotid acinar cells, NHE1 was predominant isoform in Par-C10 sells. Furthermore, functional analysis showed that Na^+/H^+ exchange in Par-C10 was as sensitive to the amiloride derivative ethylisopropylamiloride (EIPA) as the acinar cells. Taken together, our results indicate that Par-C10 cell line is a reliable model for investigating the fluid and electrolyte transport mechanism in rat parotid acinar cells.
Highly Multiplexed Fluorescence <i>in Situ</i> Hybridization for <i>in Situ</i> Genomics
Onozato, Maristela L.,Yapp, Clarence,Richardson, Douglas,Sundaresan, Tilak,Chahal, Varun,Lee, Jesse,Sullivan, James P.,Madden, Marisa W.,Shim, Hyo S.,Liebers, Matthew,Ho, Quan,Maheswaran, Shyamala,Hab American Society for Investigative Pathology 2019 The Journal of Molecular Diagnostics Vol. No.
<P>The quantification of changes in gene copy number is critical to our understanding of tumor biology and for the clinical management of cancer patients. DNA fluorescence <I>in situ</I> hybridization is the gold standard method to detect copy number alterations, but it is limited by the number of genes one can quantify simultaneously. To increase the throughput of this informative technique, a fluorescent bar-code system for the unique labeling of dozens of genes and an automated image analysis algorithm that enabled their simultaneous hybridization for the quantification of gene copy numbers were devised. We demonstrate the reliability of this multiplex approach on normal human lymphocytes, metaphase spreads of transformed cell lines, and cultured circulating tumor cells. It also opens the door to the development of gene panels for more comprehensive analysis of copy number changes in tissue, including the study of heterogeneity and of high-throughput clinical assays that could provide rapid quantification of gene copy numbers in samples with limited cellularity, such as circulating tumor cells.</P>
<i>ITGB6</i> loss-of-function mutations cause autosomal recessive amelogenesis imperfecta
Wang, Shih-Kai,Choi, Murim,Richardson, Amelia S.,Reid, Bryan M.,Lin, Brent P.,Wang, Susan J.,Kim, Jung-Wook,Simmer, James P.,Hu, Jan C.-C. Oxford University Press 2014 Human Molecular Genetics Vol.23 No.8
<P>Integrins are cell-surface adhesion receptors that bind to extracellular matrices (ECM) and mediate cell–ECM interactions. Some integrins are known to play critical roles in dental enamel formation. We recruited two Hispanic families with generalized hypoplastic amelogenesis imperfecta (AI). Analysis of whole-exome sequences identified three <I>integrin beta 6</I> (<I>ITGB6</I>) mutations responsible for their enamel malformations. The female proband of Family 1 was a compound heterozygote with an <I>ITGB6</I> transition mutation in Exon 4 (g.4545G > A c.427G > A p.Ala143Thr) and an <I>ITGB6</I> transversion mutation in Exon 6 (g.27415T > A c.825T > A p.His275Gln). The male proband of Family 2 was homozygous for an <I>ITGB6</I> transition mutation in Exon 11 (g.73664C > T c.1846C > T p.Arg616*) and hemizygous for a transition mutation in Exon 6 of <I>Nance–Horan Syndrome</I> (<I>NHS</I> Xp22.13; g.355444T > C c.1697T > C p.Met566Thr). These are the first disease-causing <I>ITGB6</I> mutations to be reported. Immunohistochemistry of mouse mandibular incisors localized ITGB6 to the distal membrane of differentiating ameloblasts and pre-ameloblasts, and then ITGB6 appeared to be internalized by secretory stage ameloblasts. ITGB6 expression was strongest in the maturation stage and its localization was associated with ameloblast modulation. Our findings demonstrate that early and late amelogenesis depend upon cell–matrix interactions. Our approach (from knockout mouse phenotype to human disease) demonstrates the power of mouse reverse genetics in mutational analysis of human genetic disorders and attests to the need for a careful dental phenotyping in large-scale knockout mouse projects.</P>
Matthew J. Garrett,R. William Richardson,Jennifer L. Wolny,B. James Williams,Michael D. Dirks,Julie A. Brame 한국조류학회I 2011 ALGAE Vol.26 No.2
Ballasting and deballasting of shipping vessels in foreign ports have been reported worldwide as a vector of introduction of non-native aquatic plants and animals. Recently, attention has turned to ballast water as a factor in the global increase of harmful algal blooms (HABs). Many species of microalgae, including harmful dinoflagellate species, can remain viable for months in dormant benthic stages (cysts) in ballast sediments. Over a period of four years, we surveyed ballast water and sediment of ships docked in two ports of Tampa Bay, Florida, USA. Sampling conditions encountered while sampling ballast water and sediments were vastly different between vessels. Since no single sample collection protocol could be applied, existing methods for sampling ballast were modified and new methods created to reduce time and labor necessary for the collection of high-quality, qualitative samples. Five methods were refined or developed,including one that allowed for a directed intake of water and sediments. From 63 samples, 1,633 dinoflagellate cysts and cyst-like cells were recovered. A native, cyst-forming, harmful dinoflagellate, Alexandrium balechii (Steidinger) F. J. R. Taylor, was collected, isolated, and cultured from the same vessel six months apart, indicating that ships exchanging ballast water in Tampa Bay have the potential to transport HAB species to other ports with similar ecologies, exposing them to non-native, potentially toxic blooms.
Garrett, Matthew J.,Wolny, Jennifer L.,Williams, B. James,Dirks, Michael D.,Brame, Julie A.,Richardson, R. William The Korean Society of Phycology 2011 ALGAE Vol.26 No.2
Ballasting and deballasting of shipping vessels in foreign ports have been reported worldwide as a vector of introduction of non-native aquatic plants and animals. Recently, attention has turned to ballast water as a factor in the global increase of harmful algal blooms (HABs). Many species of microalgae, including harmful dinoflagellate species, can remain viable for months in dormant benthic stages (cysts) in ballast sediments. Over a period of four years, we surveyed ballast water and sediment of ships docked in two ports of Tampa Bay, Florida, USA. Sampling conditions encountered while sampling ballast water and sediments were vastly different between vessels. Since no single sample collection protocol could be applied, existing methods for sampling ballast were modified and new methods created to reduce time and labor necessary for the collection of high-quality, qualitative samples. Five methods were refined or developed, including one that allowed for a directed intake of water and sediments. From 63 samples, 1,633 dinoflagellate cysts and cyst-like cells were recovered. A native, cyst-forming, harmful dinoflagellate, Alexandrium balechii (Steidinger) F. J. R. Taylor, was collected, isolated, and cultured from the same vessel six months apart, indicating that ships exchanging ballast water in Tampa Bay have the potential to transport HAB species to other ports with similar ecologies, exposing them to non-native, potentially toxic blooms.
Howard, Paul,Day, Kathleen H.,Kim, Kyoon E.,Richardson, Jeanne,Thomas, James,Abraham,Irene,Fleischmann, Robert D.,Gottesman, Michael M.,Maurer, Richard A. 충남대학교부설 생명공학연구소 1992 생물공학연구지 Vol.2 No.-
The mechansims responsible for decreased levels of cAMP-dependent protein kinase activity in a mutant Chinese hamster ovary cell line have been examined. The cAMP-resistant Chinese hamster ovary 10260 cell line was found to possess only 20% of the cAMP-dependent protein kinase activity found in wild-type cells. The pressence of decreased concentrations of the catalytic subunit in these cells was confirmed through binding studies using a radiolabeled, heat-stable inhibitor of the kinase. Cloned Chinese hamster ovary catalytic subunit cDNAs were isolated, characterized, and used as hybridization probes to examine the relative concentrations of catalytic subunit mRNAs in the wild-type and 10260 cell lines. A 40-50% decrease in the concentration of the mRNA for the Cα isozyme of the catalytic subunit was observed in 10260 cells, as compared with wild-type. This decrease in catalytic subunit mRNA concentration probably accounts for a portion of the decreased kinase activity in the mutant cell. Further analysis of Cα mRNA by polymerase chain reaction confirmed the decreased expression of Cα mRNA in 10260 cells and further demonstrated the presence of two different species of Cα cDNAs was indistinguishable from the wild-type cDNA, but the other species was shorter. Nucleotide sequence analysis of the amplified cDNAs led to the identification of a 191-base pair deletion in the shorter cDNA. Gene transfer studies using wild-type and 10260 Cα cDNAs demonstrated wild-type activity, but the shorter cDNA was inactive. These studies suggest that at least two alternations in gene expression are responsible for decreased cAMP-dependent protein kinase activity in the 10260 cell line. One alteration results in an approximately 2-fold decrease in the concentrations of Cα mRNA in the cells.