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Search Results: 1 - 10 of 323366 matches for " John J. Wille "
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Growth dynamics of individual clones of normal human keratinocytes: observations and theoretical considerations  [PDF]
John J Wille
Natural Science (NS) , 2011, DOI: 10.4236/ns.2011.38094
Abstract: The life histories of 429 individual epidermal keratinocyte clones picked at random were studied. Individual basal keratinocytes were derived from asynchronous rapidly proliferating subconfluent cultures propagated in either a low calcium (0.1mM) or a high calcium (2mM) serum-free medium. Single-celled clones were isolated by seeding trypsin-EDTA dissociated cells into a Petri dish containing cloning chips. Chips with only one cell per chip were transferred into dishes containing either low calcium or high calcium growth factor replete serum-free medium. Clone formation was monitored microscopically and the number of cells in each colony tallied at least twice daily for further analysis. A total of 369 clones were established from seven different neonatal foreskin cell strains (A-F), and 60 clones were derived from one adult human skin cell strain (G). During a five-day culture interval, among 32 clones of strain A, 83% divided at least once, 50% divided once in 24 hours, 86% divided at least three times within three days, and more than 50% divided at least four to five times in five days. Of 231 clones amongst the other five cell strains (B-F), an average of 63% (±12 S,E) divided more than three times in an eight day period, the remainder divided either once, twice or not at all. Of the 106 clones of strain G, reared in high calcium serum-free medium, 67% divided more than three times in a six-day period, and 55% divided five or more times in 6 days. Clones derived from adult skin strain H had a lower clone forming potential with 70% dividing at least once in seven days, and only 30% dividing three or more times. By contrast, the average generation time (AvGT) for second and third passage keratinocytes derived from neonatal foreskin cultures was 24 hrs. Detailed dendrograms were constructed for many of the proliferating clones. The majority of clones expressed a synblastic division pattern with every cell dividing at least once per day. A fraction of clones either exceeded this circadian division rate or displayed a biphasic division pattern with all cells initially dividing once a day and then abruptly slowing to once every other day or to an intermediate rate. A minority of clones was committed to a few terminal divisions. The division patterns of the non-synblastic clones fit an alternating bifurcated branching mode of clonal expansion expressed by the Fibonacci sequence for numbers of accumulated cells per clone per day. These results were analyzed in terms of deterministic, probabilistic and a limit cycle oscillator models of cell division timing.
Evidence for pentagonal symmetry in living and model cellular systems  [PDF]
John J. Wille
Natural Science (NS) , 2011, DOI: 10.4236/ns.2011.310112
Abstract: Microscope observations of normal human ke- ratinocytes (NHK) propagated in a serum-free medium reveal a high frequency (>70%) of pentagonally-shaped colonies over a wide range of colony sizes that persist over many sequential cell generations. NHK colonies derived from sin- gle cell isolates also display pentagonal symme- try as confirmed by a photographic technique known as “Markham Rotation”. The generality of pentagonal cellular morphology was extended to observations in situ of pentagonally-shaped basal layer epidermal cells of normal human epidermis, monolayer cultures of normal and immortalized keratinocytes, several different ch- ick embryo cells, and in previously published photographs. Statistical methods were applied that differentiate planar close-packing of polygonal configurations observed in living cellular system from several examples of non-living cellular aggregates that were produced spontaneously in nature or in the laboratory under defined physico-chemical conditions.
Occurrence of Fibonacci numbers in development and structure of animal forms: Phylogenetic observations and epigenetic significance  [PDF]
John J. Wille
Natural Science (NS) , 2012, DOI: 10.4236/ns.2012.44033
Abstract: A survey of zoological literature affirmed the wide occurrence of Fibonacci numbers in the organization of acellular and prokaryotic life forms as well as in some eukaryotic protistans and in the embryonic development and adult forms of many living and fossil remains of metazoan animals. A detailed comparative analysis of the axial skeleton of a fossil fish and humans revealed a new rule of the “nested triad” of bones organized along the proximal to distal axis of limb appendages. This growth pattern and its ubiquity among living vertebrates appear to underlie a profound rule of pattern formation that is dictated in part by the genetics and epigenetic mechanisms of stem cell clonal development.
Retinoid and Ethanol-Sensitive Benzo(α)Pyrene Induction of Cytochrome P450 in Human Keratinocytes  [PDF]
John J. Wille, Jong Y. Park
Journal of Cancer Therapy (JCT) , 2012, DOI: 10.4236/jct.2012.36141
Abstract: Polycyclic aromatic hydrocarbons (PAHs) induce cytochrome P-450 monoxygenase enzymes that catalyze the formation of DNA adducts. We investigated the effects benzo(α)pyrene (B[α]P) alone or in combination with ethanol on normal human keratinocyte (NHK) growth, induction of cytochrome P-4501A1 (CYP1A1), and modulation of these treatments by retinoic acid (RA) in a serum-free culture medium. Growth-arrested confluent NHK serum-free cultures were treated with B[α]P alone or in combination with ethanol and RA. The effects on CYP1A1 enzyme activity were investigated. B[α]P treatment alone was not toxic to post-confluent cells; sub-toxic ethanol stimulated cell growth regardless B[α]P treatment. No CYP1A1 activity was detected in control or ethanol-treated NHK cell cultures. B[α]P alone induced CYP1A1 activity, and B[α]P plus ethanol treatment further enhanced B[α]P-induced CYP1A1 activity. Pretreatment with all-trans-RA (t-RA) abolished ethanol enhancement of CYP1A1 activity. There is a synergistic action of ethanol in combination with PAH on induction of P-450 cytochrome enzymes. By contrast, RA reverses ethanol enhancement implying a role for retinoid therapy in counteracting the risk posed by combined alcohol and PAH exposure on epidermal cell carcinogenesis.
Cell Cycle Arrest Mediates Global DNA Methylation Patterns in Normal Human Keratinocytes, Epidermoid Carcinoma Cells and Murine Embryonic Fibroblasts  [PDF]
John J. Wille, Jong Y. Park
Journal of Cancer Therapy (JCT) , 2013, DOI: 10.4236/jct.2013.41030
Abstract:

The 5-methylationcytosine (5-MC) DNA content of murine embryonic fibroblasts arrested in G1 by four growth conditions (Gc, Gn, Gd, and Gs) were hypermethylated relative to rapidly growing (RG) fibroblasts. Normal human keratinocytes (NHK) arrested in G1 by suspension were hypermethylated relative to RG cultures. Four RG cultures of epidermoid carcinoma cells (ECC) were hypomethylated relative to RG NHK cultures, and two cultures (SCC25 and A431) were further hypomethylated by SUS-induced arrest. Linear regression analyses established a positive linear correlation between growth rate and 5-MC content for three murine fibroblasts lines, and a negative correlation for both NHK and ECC lines.

Effects of Okadaic Acid, Retinoic Acid, and Phorbol Myristate Acetate Tumor Promoter on Oncogene Expression  [PDF]
John J. Wille, Jong Y. Park
Journal of Cancer Therapy (JCT) , 2014, DOI: 10.4236/jct.2014.56068
Abstract:

The effect of okadaic acid (OA) on proto-oncogene protein expression of c-neu, c-myc, v-rasH, EGFR, and phosphotyrosine-containing phosphoproteins (P-Tyr) was investigated in rapidly growing (RG) normal human keratinocytes (NHK) and in SV-40 virally-transformed keratinocytes (SVK) cultured in a growth factor supplemented serum-free medium as assessed by indirect immunofluorescence microscopy. P-Tyr positively stains cell surface antigens (cytoplasm) diffusely at monopolar sites in RG NHK cultures. OA-treatment intensifies cytoplasmic P-Tyr staining at localized monopolar intercellular focal adhesion (IFA) sites with reduced cytoplasmic staining. P-Tyr expression was predominate at IFA sites with little cytoplasmic staining in RG SVK cultures. OA-treatment increased monopolar P-Tyr staining and cytoplasmic staining. OA-treatment in RG NHK cultures intensified cytoplasmic staining of c-myc and EGFR (epidermal growth factor receptor) expression. OA-treatment in RG NHK and SVK cultures intensified c-neu staining at monopolar IFA sites and intensified c-neu staining at both cytoplasmic and bipolar IFA sites in RG SVK cells. OA was especially cytotoxic for SVK cells. RA treatment decreased c-neu expression in RG NHK cultures while TPA treatment has a lesser effect on both cytoplasmic and IFA sites. RA treatment also decreased P-Tyr staining in both NHK and SVK cells. Again, TPA had a lesser inhibitory effect on P-Tyr staining pattern. RA-treatment had a similar effect on P-Tyr staining of RG cultures of a mouse fibroblast cell line. These results confirm the generality of OA, RA and TPA on the regulation of oncogene expression in both normal and malignantly transformed keratinocytes.

Induced Differentiation of Epithelioid Carcinoma Cell Lines: Evidence for Tumor Cell Quantal Mitosis  [PDF]
John J. Wille, Jong Y. Park
Journal of Cancer Therapy (JCT) , 2016, DOI: 10.4236/jct.2016.711080
Abstract: The effects of growth factors and calcium concentrations present in different culture media on induction of terminal differentiation were investigated for four different epidermoid carcinoma cell lines, Hela, KB, A431, and SCC-25, and their responses determined relative to those elicited by normal human keratinocytes subjected to these culture conditions. Differentiation status was determined cyto-chemically by a validated keratin protein staining method, and by autoradiographic analyses. Growth and differentiation promoting factors that influenced the direction of integrated control of growth and differentiation in normal human keratinocytes were found to be effective for some cell lines but not others. The factors examined were 1) high density arrest in serum-free and serum-containing media, 2) media shifts from high density culture in serum-containing media to low density growth factor-depleted or supplemented serum-free medium, and 3) the concentration of calcium in the media. The extent and degree of differentiation achieved varied among different cell lines depend on the presence or absence of serum, EGF and insulin protein growth factors. Certain growth media appear to sponsor keratin protein, cyto-chemically-detected differentiation, and evidence of quantal mitotic division in low density HeLa cell and SCC25 cell cultures. Epidermoid carcinoma cell lines retain limited capacity to commit to early stages of cell differentiation.
Cancer Chemopreventive Retinoids: Validation and Analysis of in Vivo and in Vitro Bioassay Results  [PDF]
John J. Wille, Jong Y. Park, Y. Fulmer Shealy
Journal of Cancer Therapy (JCT) , 2016, DOI: 10.4236/jct.2016.713098
Abstract: Several natural and synthetic retinoids (vitamin-A derived analogies) were examined for their potential anti-cancer activity in both in vivo animal models and a novel in vitro human keratinocyte clonal growth bioassay system. The natural retinoids included all-trans-retinoic (RA), 13-cis-retinoic acid, 4-oxoretinoic acid, and retinol. Among the synthetic retinoids tested were all trans N-(4-hydroxy(phenyl)retinamide, 3-substituted oxoretinoic acids, and 13 cis-N-ethylretinamide. The animal models employed were: 1) vitamin A-deficient hamster tracheal organ assay (HTOC); 2) the benzo(α)pyrene-induced squamous metaplasia in a hamster tracheal organ system (BP-HTOC); 3) the mouse skin tumor promoter (TPA)-induced ornithine decarboxylase enzyme assay(ODC); 4) the mouse skin papilloma (MPA) assay; and 5) a novel retinoid bioassay in which retinoids display IC50 values to inhibit clonal growth of NHK. All-trans-RA, 4-oxoretinoic acid and retinol were consistently more active than any of the synthetic derivatives in all bioassays tested. A statistical model was developed and significant positive correlations were found between: 1) ED50 values in the HTOC system and reduction in TPA-induced ODC enzyme activity; 2) tumors per animal in the MPA bioassay and suppression of TPA-induced ODC activity; and 3) a positive correlation between suppression of tumors per animal in the MPA assay, and retinoid inhibition of keratinocyte clonal growth. Test retinoids, were tested for their capacity to inhibit the clonal growth of a squamous carcinoma cell line (SCC-25), which were found to be 2
Differential Anticancer Effect of an Apple Extract (Applephenon®), Polyphenols and Isoflavones on Normal Human Keratinocytes and Epidermoid Cancer Cells  [PDF]
John J. Wille, Mark A. Berhow, Jong Y. Park
Journal of Cancer Therapy (JCT) , 2019, DOI: 10.4236/jct.2019.106040
Abstract: Applephenon®, a purified extract prepared from green apples, was examined for its cytotoxicity and inhibitory effects on the proliferation of cultures of normal human keratinocytes and several epidermoid cancer cell lines. Our HPLC studies demonstrated a high content of phenolic compounds (>65%), including catechin, epicatechin, caffeic acid and phloretin as well as polyphenols such as proanthocyanidins. Applephenon® demonstrated a greater cytotoxic effect against HeLa, A431 cancer cell lines and HaCaT, an immortalized keratinocyte cell line than serum-free cultures of proliferating normal human keratinocytes (NHK). Proliferation of NHK was inhibited at concentrations above 0.0013% while concentrations above 0.005% were cytotoxic. By contrast, Applephenon® solutions above 0.00025% killed each of the cancer cell lines. Treated cells displayed increased intercellular separation and evidence of keratinizing stratification. We also tested the effect of epicatechin, and two isoflavonoids, genistein and daidzein, on cancer cell lines. Hela cells were more sensitive to epicatechin and genistein inhibition of cell growth and cytotoxicity than were NHK. Daidzein at these concentrations had little effect on cancer cells. These results indicate that Applephenon® and some of its phenolic components have selective anticancer activity.
Probing the Nucleation of Al2O3 in Atomic Layer Deposition on Aluminum for Ultrathin Tunneling Barriers in Josephson Junctions
Alan J. Elliot,Gary Malek,Logan Wille,Rongtao Lu,Siyuan Han,Judy Z. Wu,John Talvacchio,Rupert M. Lewis
Physics , 2013, DOI: 10.1109/TASC.2013.2247452
Abstract: Ultrathin dielectric tunneling barriers are critical to Josephson junction (JJ) based superconducting quantum bits (qubits). However, the prevailing technique of thermally oxidizing aluminum via oxygen diffusion produces problematic defects, such as oxygen vacancies, which are believed to be a primary source of the two-level fluctuators and contribute to the decoherence of the qubits. Development of alternative approaches for improved tunneling barriers becomes urgent and imperative. Atomic Layer Deposition (ALD) of aluminum oxide (Al2O3) is a promising alternative to resolve the issue of oxygen vacancies in the Al2O3 tunneling barrier, and its self-limiting growth mechanism provides atomic-scale precision in tunneling barrier thickness control. A critical issue in ALD of Al2O3 on metals is the lack of hydroxyl groups on metal surface, which prevents nucleation of the trimethylaluminum (TMA). In this work, we explore modifications of the aluminum surface with water pulse exposures followed by TMA pulse exposures to assess the feasibility of ALD as a viable technique for JJ qubits. ALD Al2O3 films from 40 angstroms to 100 angstoms were grown on 1.4 angstroms to 500 angstroms of Al and were characterized with ellipsometry and atomic force microscopy. A growth rate of 1.2 angstroms/cycle was measured, and an interfacial layer (IL) was observed. Since the IL thickness depends on the availability of Al and saturated at 2 nm, choosing ultrathin Al wetting layers may lead to ultrathin ALD Al2O3 tunneling barriers.
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