SPRR4, a novel cornified envelope precursor: UV-dependent epidermal expression and selective incorporation into fragile envelopes

The cornified cell envelope (CE), a structure formed in the outermost layers of stratified squamous epithelia, provides a physical barrier against environmental insults. It is composed of several structural proteins, which are irreversibly crosslinked by calcium-activated transglutaminases. The small proline rich proteins (SPRRs) are one set of CE precursors. SPRR4, a novel member of this gene family, displayed very low or undetectable expression levels in normal human skin or other stratified squamous epithelia, but was clearly induced by UV light both in vivo and in vitro. High epidermal expression of SPRR4 was monitored only after chronic UV exposure and was concomitant with a thickening of the stratum corneum, which is believed to provide protection against subsequent damage. The calcium-dependent translocation of an SPRR4-GFP fusion protein to the cell periphery in living keratinocytes and its integration into both rigid and fragile cornified envelopes proved that SPRR4 is a novel CE precursor. Interestingly, after UV irradiation, SPRR4 was selectively incorporated into fragile CEs. Our results show for the first time that UV-induced cornification is accompanied by qualitative changes in CE precursor assembly. SPRR4 is part of an adaptive tissue response to environmental stress, which is likely to compensate for UV induced impairment of the epidermal barrier function.

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Optimization of zinc oxide nanoparticle-catalyzed in vitro bilirubin photolysis and in vivo treatment of hyperbilirubinemia

Nanomedicine ◽

10.2217/nnm-2021-0036 ◽

2021 ◽

Author(s):

Haq Nawaz ◽

Iqra Naseem ◽

Tanzila Rehman ◽

Mubashir Nawaz

Keyword(s):

Zinc Oxide ◽

Zinc Oxide Nanoparticles ◽

Uv Light ◽

Uv Exposure ◽

Uv Treatment ◽

Blood Samples ◽

Positive Effects ◽

Oxide Nanoparticle

Aim: To optimize the Zinc oxide nanoparticles (ZnONPs)-catalyzed in vitro photolysis of bilirubin and to test their effect on bilirubin clearance in vivo. Materials & methods: ZnONPs, synthesized in an alkaline medium, were characterized. Response surface methodology was used to optimize the in vitro photolysis catalyzed by the nanoparticles (NPs). Blood samples from phenylhydrazine-induced hyperbilirubinemic rabbits which had been administered ZnONPs and UV light were analyzed to assess in vivo clearance of bilirubin. Results: The ZnONP-assisted UV treatment showed the linear and quadratic positive effects on the in vitro bilirubin photolysis with an optimal photolysis of bilirubin at 225 mg dl-1 concentration of ZnONPs and a UV exposure of 1.80 h. The ZnONP-assisted phototherapy of hyperbilirubinemic animals was also found to be more effective for in vivo clearance of bilirubin than phototherapy alone. Conclusion: After further trials, ZnONP-assisted phototherapy could be a potential treatment for hyperbilirubinemia in humans.

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Regulation of SH3PX1 by dNedd4-long at the Drosophila neuromuscular junction

Journal of Biological Chemistry ◽

10.1074/jbc.ra118.005161 ◽

2018 ◽

Vol 294 (5) ◽

pp. 1739-1752 ◽

Cited By ~ 2

Author(s):

Samantha S. Wasserman ◽

Alina Shteiman-Kotler ◽

Kathryn Harris ◽

Konstantin G. Iliadi ◽

Avinash Persaud ◽

...

Keyword(s):

Neurotransmitter Release ◽

Neuromuscular Junctions ◽

Cell Periphery ◽

Middle Region ◽

Binding Partners ◽

Direct Binding ◽

Specific Regulation ◽

Neuromuscular Synaptogenesis

Drosophila Nedd4 (dNedd4) is a HECT E3 ubiquitin ligase present in two major isoforms: short (dNedd4S) and long (dNedd4Lo), with the latter containing two unique regions (N terminus and Middle). Although dNedd4S promotes neuromuscular synaptogenesis (NMS), dNedd4Lo inhibits it and impairs larval locomotion. To explain how dNedd4Lo inhibits NMS, MS analysis was performed to find its binding partners and identified SH3PX1, which binds dNedd4Lo unique Middle region. SH3PX1 contains SH3, PX, and BAR domains and is present at neuromuscular junctions, where it regulates active zone ultrastructure and presynaptic neurotransmitter release. Here, we demonstrate direct binding of SH3PX1 to the dNedd4Lo Middle region (which contains a Pro-rich sequence) in vitro and in cells, via the SH3PX1-SH3 domain. In Drosophila S2 cells, dNedd4Lo overexpression reduces SH3PX1 levels at the cell periphery. In vivo overexpression of dNedd4Lo post-synaptically, but not pre-synaptically, reduces SH3PX1 levels at the subsynaptic reticulum and impairs neurotransmitter release. Unexpectedly, larvae that overexpress dNedd4Lo post-synaptically and are heterozygous for a null mutation in SH3PX1 display increased neurotransmission compared with dNedd4Lo or SH3PX1 mutant larvae alone, suggesting a compensatory effect from the remaining SH3PX1 allele. These results suggest a post-synaptic–specific regulation of SH3PX1 by dNedd4Lo.

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NO modulates myocardial O2consumption in the nonhuman primate: an additional mechanism of action of amlodipine

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1999.276.6.h2069 ◽

1999 ◽

Vol 276 (6) ◽

pp. H2069-H2075 ◽

Cited By ~ 7

Author(s):

Paul R. Forfia ◽

Xiaoping Zhang ◽

Delvin R. Knight ◽

Andrew H. Smith ◽

Christopher P. A. Doe ◽

...

Keyword(s):

Nitric Oxide ◽

Oxygen Consumption ◽

Nonhuman Primate ◽

Myocardial Oxygen Consumption ◽

Channel Blocker ◽

Tissue Response ◽

Canine Heart ◽

Additional Mechanism

Recent evidence from our laboratory and others suggests that nitric oxide (NO) is a modulator of in vivo and in vitro oxygen consumption in the murine and canine heart. Therefore, the goal of our study was twofold: to determine whether NO modulates myocardial oxygen consumption in the nonhuman primate heart in vitro and to evaluate whether the seemingly cardioprotective actions of amlodipine may involve an NO-mediated mechanism. Using a Clark-type O2 electrode, we measured oxygen consumption in cynomologous monkey heart at baseline and after increasing doses of S-nitroso- N-acetylpenicillamine (SNAP; 10−7–10−4M), bradykinin (10−7–10−4M), ramiprilat (10−7–10−4M), and amlodipine (10−7–10−5M). SNAP (−38 ± 5.8%), bradykinin (−19 ± 3.9%), ramiprilat (−28 ± 2.3%), and amlodipine (−23 ± 4.5%) each caused significant ( P < 0.05) reductions in myocardial oxygen consumption at their highest dose. Preincubation of tissue with nitro-l-arginine methyl ester (10−4 M) blunted the effects of bradykinin (−5.4 ± 3.2%), ramiprilat (−4.8 ± 5.0%), and amlodipine (−5.3 ± 5.0%) but had no effect on the tissue response to SNAP (−38 ± 5.8%). Our results indicate that NO can reduce oxygen consumption in the primate myocardium in vitro, and they support a role for the calcium-channel blocker amlodipine as a modulator of myocardial oxygen consumption via a kinin-NO mediated mechanism.

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Studies on the form and synthesis of messenger ribonucleic acid in the rat ventral prostate gland, including its tissue-specific stimulation by androgens

Biochemical Journal ◽

10.1042/bj1370513 ◽

1974 ◽

Vol 137 (3) ◽

pp. 513-524 ◽

Cited By ~ 27

Author(s):

W. Ian P. Mainwaring ◽

Peter A. Wilce ◽

Allan E. Smith

Keyword(s):

Prostate Gland ◽

Sodium Dodecyl ◽

Ventral Prostate ◽

Experimental Conditions ◽

Free System ◽

Cell Free System ◽

Messenger Ribonucleic Acid ◽

Selective Incorporation

1. When prostate polyribosomes are labelled with radioactive precursors in vivo and subsequently dissociated with sodium dodecyl sulphate, a heterogeneous 6–15S RNA species may be identified that possesses all of the distinctive properties of mRNA. 2. Apart from the selective incorporation of 5′-fluoro-orotic acid into this 6–15S RNA component, it is bound by nitrocellulose filters under experimental conditions where only poly(A)-rich species of RNA are specifically retained. Most importantly, however, only the 6–15S RNA fraction is capable of promoting the incorporation of amino acids into peptide linkage in an mRNA-depleted cell-free system derived from ascites-tumour cells. 3. With the development of a simpler method for labelling the total RNA fraction of the prostate gland in vitro, the poly(A)-enriched RNA fraction may be readily isolated by adsorption and elution from oligo(dT)-cellulose. The synthesis of the poly(A)-enriched 6–15S RNA fraction is stringently controlled by androgens in a highly tissue- and steroid-specific manner. 4. From an analysis of the proteins synthesized in the ascites cell-free system in the presence of the poly(A)-rich RNA fraction, it appears that protein synthesis in the prostate gland is stimulated in a rather general way, even during the earliest phases of the androgenic response. This conclusion may require modification when more specific means of analysis are available than those used in the present investigation. 5. The implications of these findings to the mechanism of action of androgens are discussed.

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Schistosoma mansoni in Susceptible and Resistant Snail Strains Biomphalaria tenagophila: In Vivo Tissue Response and In Vitro Hemocyte Interactions

PLoS ONE ◽

10.1371/journal.pone.0045637 ◽

2012 ◽

Vol 7 (9) ◽

pp. e45637 ◽

Cited By ~ 13

Author(s):

Rafael Nacif-Pimenta ◽

Ana Carolina Alves de Mattos ◽

Alessandra da Silva Orfanó ◽

Luciene Barbosa ◽

Paulo Filemon Paolucci Pimenta ◽

...

Keyword(s):

Schistosoma Mansoni ◽

Tissue Response

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136 IN VITRO DEVELOPMENT OF BOVINE EMBRYOS SUBMITTED TO LASER ASSISTED HATCHING ON DAY 6 AND DAY 7 OF EMBRYO CULTURE

Reproduction Fertility and Development ◽

10.1071/rdv21n1ab136 ◽

2009 ◽

Vol 21 (1) ◽

pp. 168

Author(s):

C. B. Ponchirolli-Schneider ◽

J. H. Pryor ◽

C. R. Looney

Keyword(s):

Embryo Culture ◽

Uv Light ◽

Laser System ◽

Pulse Length ◽

Assisted Hatching ◽

In Vitro Production ◽

Stage Of Development ◽

Number Of Cells

In vitro production (IVP) of embryos is a valuable tool in bovine assisted reproduction. IVP embryos show lower pregnancy rates when compared to in vivo produced embryos. The inability of the IVP embryo to hatch from the zona pellucida (ZP) after embryo transfer is believed to be one contributing factor. The aim of this study was to compare the ability of IVP embryos to hatch and develop in vitro after being submitted to laser assisted hatching (LAH) at two different time periods of embryo culture: 144 h (Day 6) and 168 h (Day 7). In vitro maturated oocytes from slaughterhouse ovaries (Ovitra Biotechnologies, Amarillo, TX, USA) were fertilized with frozen/thawed semen from two different bulls (Day 0) and cultured in G1.5/G2.5 medium supplemented with 8 mg mL–1 of BSA (Vitrolife, Englewood, CA, USA) at 38.5°C, 5% CO2, 5% O2, 5% N2, in humidified atmosphere. On Day 6 post-fertilization, all viable embryos (n = 251), from three replicates, were washed in holding medium (Vigro Holding Plus, Bioniche, Pullman, WA, USA) and divided into four treatment groups: laser assisted hatching Day 6 and Day 7 (LAHD6 and LAHD7), control Day 6 and Day 7 (CD6 and CD7). The groups LAHD7 and CD7 were immediately placed in G2.5 and returned to the incubator until Day 7. Embryos from groups LAHD6 and CD6 were placed in G2.5 in separate wells of a four well dish and covered with 300 μL of mineral oil. Embryos from LAHD6 group had one third to one half of the external edge of the ZP exposed to a laser beam, using XY Clone® (Hamilton Thorne Biosciences, Inc., Beverly, MA, USA) laser system, with a pulse strength of 90% and a pulse length of 600 μs. The group CD6 was submitted to the same conditions, but did not receive the laser treatment. On Day 7, the experiment was repeated for embryos belonging to groups LAHD7 and CD7. Embryos from all groups were cultured in vitro and evaluated on Day 8 and Day 9 for stage of development. On Day 9, a random sample of embryos from each treatment group (n = 48) was stained with Hoechst 33342 (2.5 μg mL–1) and evaluated under UV light to determine the total number of cells. The number of hatched blastocysts was not different (chi-square, p > 0.05) among the groups on Day 9 of culture (LAHD6 = 49/66, CD6 = 38/61, LAHD7 = 42/59, CD7 = 47/65; 74, 62, 71 and 72%, respectively). However, on Day 8 of culture, LAHD6 showed a higher number (p < 0.05) of hatched blastocysts (40/66, 60%), when compared to group CD6 (26/61, 42%). There was no difference between groups LAHD7 (33/59, 55%) and CD7 (31/65, 47%) on Day 8. Comparison of the total number of cells showed no difference (Student’s t-test, p > 0.05) among the groups (LAHD6 = 154.8 ± 12.2, CD6 = 184.4 ± 19.5, LAHD7 = 170.4 ± 15.8, CD7 = 162.5 ± 14.6), indicating that LAH does not have a detrimental effect on mean cell production throughout embryo development in vitro. These data show that LAH on Day 6 of culture improves in vitro hatching rates on Day 8, while LAH on Day 7 shows no improvement on either Day 8 or 9. Kathy Bradley, Hamilton Thorne Biosciences, Inc.

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Early Biofilm Formation on UV Light Activated Nanoporous TiO2SurfacesIn Vivo

International Journal of Biomaterials ◽

10.1155/2018/7275617 ◽

2018 ◽

Vol 2018 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Nagat Areid ◽

Eva Söderling ◽

Johanna Tanner ◽

Ilkka Kangasniemi ◽

Timo O. Närhi

Keyword(s):

Titanium Alloy ◽

Biofilm Formation ◽

Uv Light ◽

Antibacterial Properties ◽

Mutans Streptococci ◽

Human Gingival Fibroblast ◽

Gingival Fibroblast ◽

Biofilm Development

Purpose. To explore earlyS. mutansbiofilm formation on hydrothermally induced nanoporous TiO2surfacesin vivoand to examine the effect of UV light activation on the biofilm development.Materials and Methods. Ti-6Al-4V titanium alloy discs (n = 40) were divided into four groups with different surface treatments: noncoated titanium alloy (NC); UV treated noncoated titanium alloy (UVNC); hydrothermally induced TiO2coating (HT); and UV treated titanium alloy with hydrothermally induced TiO2coating (UVHT).In vivoplaque formation was studied in 10 healthy, nonsmoking adult volunteers. Titanium discs were randomly distributed among the maxillary first and second molars. UV treatment was administered for 60 min immediately before attaching the discs in subjects’ molars. Plaque samples were collected 24h after the attachment of the specimens. Mutans streptococci (MS), non-mutans streptococci, and total facultative bacteria were cultured, and colonies were counted.Results. The plaque samples of NC (NC + UVNC) surfaces showed over 2 times more oftenS. mutanswhen compared to TiO2surfaces (HT + UVHT), with the number of colonized surfaces equal to 7 and 3, respectively.Conclusion. Thisin vivostudy suggested that HT TiO2surfaces, which we earlier showed to improve blood coagulation and encourage human gingival fibroblast attachmentin vitro, do not enhance salivary microbial (mostly mutans streptococci) adhesion and initial biofilm formation when compared with noncoated titanium alloy. UV light treatment provided Ti-6Al-4V surfaces with antibacterial properties and showed a trend towards less biofilm formation when compared with non-UV treated titanium surfaces.

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Cumulus Cell Expansion, Its Role in Oocyte Biology and Perspectives of Measurement: A Review

Scientia Agriculturae Bohemica ◽

10.1515/sab-2015-0002 ◽

2015 ◽

Vol 45 (4) ◽

pp. 212-225 ◽

Cited By ~ 4

Author(s):

J. Nevoral ◽

M. Orsák ◽

P. Klein ◽

J. Petr ◽

M. Dvořáková ◽

...

Keyword(s):

Extracellular Matrix ◽

Hyaluronic Acid ◽

High Performance ◽

Uv Light ◽

Cumulus Cell ◽

Developmental Competence ◽

Enzyme Linked Immunosorbent Assay ◽

Cumulus Expansion

Abstract Cumulus expansion of the cumulus-oocyte complex is necessary for meiotic maturation and acquiring developmental competence. Cumulus expansion is based on extracellular matrix synthesis by cumulus cells. Hyaluronic acid is the most abundant component of this extracellular matrix. Cumulus expansion takes place during meiotic oocyte maturation under in vivo and in vitro conditions. Quantification and measurement of cumulus expansion intensity is one possible method of determining oocyte quality and optimizing conditions for in vitro cultivation. Currently, subjective methods of expanded area and more exact cumulus expansion measurement by hyaluronic acid assessment are available. Among the methods of hyaluronic acid measurement is the use of radioactively labelled synthesis precursors. Alternatively, immunological and analytical methods, including enzyme-linked immunosorbent assay (ELISA), spectrophotometry, and high-performance liquid chromatography (HPLC) in UV light, could be utilized. The high sensitivity of these methods could provide a precise analysis of cumulus expansion without the use of radioisotopes. Therefore, the aim of this review is to summarize and compare available approaches of cumulus expansion measurement, respecting special biological features of expanded cumuli, and to suggest possible solutions for exact cumulus expansion analysis.

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Testing of resveratrol microemulsion photostability and protective effect against UV induced oxidative stress

Acta Pharmaceutica ◽

10.1515/acph-2017-0018 ◽

2017 ◽

Vol 67 (2) ◽

pp. 247-256 ◽

Cited By ~ 8

Author(s):

Vaida Juškaitė ◽

Kristina Ramanauskienė ◽

Vitalis Briedis

Keyword(s):

Oxidative Stress ◽

Antioxidant Activity ◽

Ultraviolet Radiation ◽

Uv Light ◽

Biological Testing ◽

Human Keratinocyte ◽

Improved Stability ◽

Rate Profile

Abstract Resveratrol is well known for its antioxidant activity and susceptibility to ultraviolet radiation. Development of formulations providing improved stability and relevant drug delivery of resveratrol is still a challenging task. The aim of this study was to determine protective characteristics of formulated microemulsions by evaluating photoisomerization of resveratrol and to investigate the effects of resveratrol on human keratinocyte cells under oxidative stress caused by ultraviolet radiation. Incorporation of resveratrol into microemulsions resulted in increased photostability of active compounds and the results demonstrated that photodegradation of resveratrol was significantly delayed. Results of biopharmaceutical evaluation in vitro demonstrated that up to 60 % of resveratrol was released from microemulsions within 6 hours under a constant release rate profile. In vivo biological testing confirmed the ability of resveratrol to protect cells from oxidative stress and to increase cell viability. It was concluded that microemulsions might be considered in the development of UV light sensitive compounds.

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