FIGURE 4 - uploaded by Martin Schicht
Content may be subject to copyright.
In vitro stimulation with bacterial supernatants. Real-time RT-PCR of PLUNC: Stimulation of HCE cells (A) and HCjE cells (C) for 6, 24, and 48 hours with bacterial supernatant of Escherichia coli (E. coli) and Pseudomonas aeruginosa (PA). ELISA of PLUNC: Stimulation of HCE cells (B) and HCjE cells (D) for 6, 24, and 48 hours with bacterial supernatant of E. coli and PA. Statistical significance (n ¼ 3; *P 0.05). The regulation of PLUNC transcript levels was expressed as mean 6 SEM.
Source publication
Purpose:
Palate Lung Nasal Clone (PLUNC) is a hydrophobic protein belonging to the family of surfactant proteins that is involved in fluid balance regulation of the lung. Moreover, it is known to directly act against gram-negative bacteria. The purpose of this study was to investigate the possible expression and antimicrobial role of PLUNC at the...
Contexts in source publication
Context 1
... cornea epithelial (HCE) and conjunctivial epithelial cell line (HCjE) were used and cultivated to investigate whether PLUNC is regulated by proinflammatory cytokines and bacterial supernatants. Both cell lines are well established and known to produce surfactant proteins after stimulation with bacterial supernatants. After stimulation with IL-1b or TNFa, an upregulation of PLUNC expression could be observed in HCE after real-time PCR analysis (Fig. 3C). Stimulation with LPS revealed no significant effect. In contrast, stimulation with LPS showed a significantly upregulation in HCjE, whereas in this cell line, IL-1b and TNFa demonstrated no effect (Fig. 3D). A significant upregulation in the PLUNC mRNA expression could be detected after stimulation with bacterial supernatants of EC, but not with PA in HCE cells (Fig. 4A). The PA had a slight effect on the PLUNC expression after 6-hour stimulation time in HCjE cells, whereas EC had no effect in this case (Fig. 4B). Enzyme-linked immunosorbent assay indicated that none of the stimulants (IL-1b, LPS, TNFa, EC, PA) had any significant effect on the PLUNC protein concentration in HCE cells (Figs. 3D, 4C). In HCjE cells, IL-1b and TNFa significantly reduced the PLUNC concentration over time, whereas LPS and supernatants of EC and PA had no effect (Figs. 3F, ...
Context 2
... cornea epithelial (HCE) and conjunctivial epithelial cell line (HCjE) were used and cultivated to investigate whether PLUNC is regulated by proinflammatory cytokines and bacterial supernatants. Both cell lines are well established and known to produce surfactant proteins after stimulation with bacterial supernatants. After stimulation with IL-1b or TNFa, an upregulation of PLUNC expression could be observed in HCE after real-time PCR analysis (Fig. 3C). Stimulation with LPS revealed no significant effect. In contrast, stimulation with LPS showed a significantly upregulation in HCjE, whereas in this cell line, IL-1b and TNFa demonstrated no effect (Fig. 3D). A significant upregulation in the PLUNC mRNA expression could be detected after stimulation with bacterial supernatants of EC, but not with PA in HCE cells (Fig. 4A). The PA had a slight effect on the PLUNC expression after 6-hour stimulation time in HCjE cells, whereas EC had no effect in this case (Fig. 4B). Enzyme-linked immunosorbent assay indicated that none of the stimulants (IL-1b, LPS, TNFa, EC, PA) had any significant effect on the PLUNC protein concentration in HCE cells (Figs. 3D, 4C). In HCjE cells, IL-1b and TNFa significantly reduced the PLUNC concentration over time, whereas LPS and supernatants of EC and PA had no effect (Figs. 3F, ...
Similar publications
Background:
International Task Force (ITF) guidelines established a grading scheme to support treatment of dry eye disease based on clinical signs and symptoms. The purpose of this study was to assess the impact of dry eye on vision-related function across ITF severity levels using the Ocular Surface Disease Index (OSDI) questionnaire.
Methods:...
Dry eye disease (DED) is a chronic and progressive multifactorial disorder of the tears and ocular surface, which results in symptoms of discomfort and visual disturbance. The aim of this systematic literature review was to evaluate the burden of DED and its components from an economic and health-related quality of life (HRQoL) perspective, and to...
To investigate the effects of polyunsaturated fatty acids (PUFAs) in patients with dry eye disease (DED), a multifactorial inflammatory disorder, we searched Cochrane Library, EMBASE, PubMed, and Web of Science for randomized clinical trials (RCTs) investigating the effect of PUFAs in patients with DED before March 2019. Two reviewers independently...
p> Background: Diabetes is one of the most common leading causes of blindness in 20–74 year old persons. Recently, problems involving the ocular surface, dry eyes in particular, have been reported in diabetic patients. This study was performed to assess the prevalence of dry eyes syndrome and diabetic retinopathy and its association with HbA1c in d...
![FIGURE 5. Correlations between cytokines (interleukin [IL]-17A and...](profile/Melissa-Toyos/publication/319574711/figure/fig3/AS:675469112401923@1538055794524/Correlations-between-cytokines-interleukin-IL-17A-and-IL-6-and-ocular-surface_Q320.jpg)
Purpose:
To investigate the change from baseline of inflammatory markers in tears of dry eye disease(DED) subjects due to meibomian gland dysfunction(MGD) after IPL(intense pulsed light) treatment and meibomian gland expression(MGE) compared to sham treatment, and the correlations with ocular surface parameters.
Design:
Randomized, double-masked...
Citations
... However, due to the lack of knowledge about the function of SP-G, this finding can only be speculated. However, a correspondingly higher concentration in the cornea was also detected for the protein PLUNC (Palate Lung Nasal Clone), which belongs to the family of surfactant proteins [25]. Here, it is speculated that PLUNC may have a function in the cornea, which is an organ with a very finely regulated fluid balance, in the context of fluid regulation or in immune defense. ...
... The procedure of collecting tear fluid as well as the clinical diagnosis of dry eye has already been described and performed in detail by Schicht et al. [25]. Tear fluid samples were collected with Schirmer strips through PD (Dr. ...
... Immortalized HCE and HCjE cells were cultured as previously described by [18]. SV40transformed human corneal epithelial cells (HCE cells, obtained from Kaoru Araki-Sasaki, Tane Memorial Eye Hospital, Osaka, Japan, passage number [18][19][20][21][22][23][24][25][26][27] [36] as well as a human spontaneously immortalized epithelial cell line from normal human conjunctiva [IOBA-NHC, here referred to as HCjE cells, obtained from Yolanda Diebold, University Institute of Applied Ophthalmobiology (IOBA), University of Valladolid, Valladolid, Spain] [37] were cultured as monolayer and used for further stimulation experiments. For stimulation experiments, cells (1 × 10 6 ) were seeded in Petri dishes and cultured until confluence was reached. ...
The ocular surface is in constant interaction with the environment and with numerous pathogens. Therefore, complex mechanisms such as a stable tear film and local immune defense mechanisms are required to protect the eye. This study describes the detection, characterization, and putative role of surfactant protein G (SP-G/SFTA2) with respect to wound healing and surface activity. Bioinformatic, biochemical, and immunological methods were combined to elucidate the role of SP-G in tear film. The results show the presence of SP-G in ocular surface tissues and tear film (TF). Increased expression of SP-G was demonstrated in TF of patients with dry eye disease (DED). Addition of recombinant SP-G in combination with lipids led to an accelerated wound healing of human corneal cells as well as to a reduction of TF surface tension. Molecular modeling of TF suggest that SP-G may regulate tear film surface tension and improve its stability through specific interactions with lipids components of the tear film. In conclusion, SP-G is an ocular surface protein with putative wound healing properties that can also reduce the surface tension of the tear film.
... Thirdly, urea has the capacity to induce conformation changes in proteins and peptides (Caballero-Herrera et al., 2005). This may also include surface-active proteins like surfactant proteins (SP)-A, SP-B, SP-C, SP-D, and SP-H or associated proteins such as PLUNC (Palate Lung Nasal Clone), all of which have been detected at the ocular surface and in tear fluid (Bräuer et al., 2007a(Bräuer et al., , 2007bSchicht et al., 2018Schicht et al., , 2015. Consequently, it is very likely that urea indirectly affects the surface tension of the tear film. ...
Objective
Urea is a component of tear fluid showing a significantly decreased concentration in dry eye disease. The urea content of tear fluid may depend on urea transporters. The purpose of this study was to examine the expression of urea transporter B (UT-B) at the ocular surface and in the lacrimal glands.
Methods
UT-B protein and mRNA expression was investigated in human, porcine, and murine samples. Immunohistochemical staining for UT-B was performed on paraffin sections of human, porcine, and murine corneas, eyelids, and lacrimal glands (n=5 each). Reverse transcriptase polymerase chain reaction was conducted to detect UT-B mRNA in human and murine cornea, conjunctiva, Meibomian gland, and lacrimal gland (n=5 each).
Results
UT-B protein expression was comparable in all three species. It was found in the corneal epithelium and endothelium, in the conjunctival epithelium, in the end pieces and excretory ducts of the lacrimal gland, Meibomian gland, and in the glands of Moll and Zeis. The glands of Zeis and the Meibomian glands showed intense UT-B signals in the basal layers of the alveolar epithelia and in the cells of the ductal epithelia. UT-B mRNA was detected in all samples analyzed.
Conclusion
UT-B is expressed by the cells and tissues of the ocular surface and in the lacrimal glands. Potential changes in urea transporter expression might have implications for the pathogenesis of dry eye disease. Since comparable results were obtained for all species investigated, the presented findings may open the door for DED-relevant experimentation on porcine and murine model systems.
... Each sample (30 µg protein) was mixed with reducing sample buffer (RSB) containing 0.5 M dithiothreitol (DTT), 50% glycerin, 0.05% bromphenolblau, 20% β-mercaptoethanol (β-ME) and boiled for 5 minutes. SDS-PAGE and Western blot analyses were performed as described by [56]. The membrane was incubated with primary antibodies to TFF1 (dilution 1:100), TFF2 (dilution 1:70), and TFF3 (dilution 1:400) at 4 • C overnight before applying the secondary antibodies (dilution 1:4000) conjugated to horseradish peroxidase for 2 h and detecting the bands by chemiluminescence using ECL Plus (Amersham Pharmacia, Uppsala, Sweden). ...
Hintergrund und Ziele: Trefoil factor family Peptide (TFF) sind eine Gruppe von drei (TFF1, TFF2 und TFF3) kleeblattförmigen Peptiden, die in verschiedenen Geweben und Flüssigkeiten des menschlichen Körpers vorkommen. Hier interagieren sie mit unterschiedlichen Molekülen und Rezeptoren, u.a. mit Muzinen und Chemokinrezeptoren (Braga Emidio et al., 2019, Otto and Thim, 2005). Den TFF-Peptiden werden protektive und wundheilungsfördernde Eigenschaften z.B. in Schleimhäuten zugeschrieben (Taupin and Podolsky, 2003); auf der anderen Seite wurde aber auch gezeigt, dass das Trefoil factor family Peptid 3 (TFF3) im arthrotischen Gelenkknorpel katabole Eigenschaften aufweist (Rösler et al., 2010). Rheumatoide Arthritis (RA) und Arthrose (OA) stellen zwei der häufigsten Gelenkerkrankungen dar, die für die betroffenen Patient*innen mit hohem Leidensdruck aufgrund von Schmerzen und Gelenkdeformitäten einhergehen. Beide Erkrankungen sind durch Entzündungsvorgänge und Destruktionen im beteiligten Gelenkknorpel gekennzeichnet. Die Vorgänge betreffen nicht nur isoliert den Gelenkknorpel, sondern auch die umliegenden Gelenkstrukturen, wie die Synovialmembran (SM) und die Synovialflüssigkeit (SF, Synovia). Ziel dieser Arbeit ist es zu analysieren, ob die TFF-Peptide 1-3 ebenfalls von Zellen der SM produziert werden und in der SF vorkommen. Hierbei werden Proben von gesunden Proband*innen mit denen von Patient*innen, die an OA oder RA leiden, verglichen. Methoden: Zur Detektion, Lokalisation und Quantifizierung der TFF-Peptide werden methodisch reverse Transkriptase Polymerase-Kettenreaktion (RT-PCR), real-time RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay (ELISA) und Immunhistochemie angewandt. Für die Untersuchungen werden hierbei Gewebe und Gelenkflüssigkeit gelenkgesunder Personen sowie von Patient*innen mit OA und RA unterschiedlichen Alters und Geschlechts eingesetzt und miteinander verglichen. Ergebnisse und Beobachtungen: Die Ergebnisse zeigen, dass in SM ausschließlich TFF3 exprimiert und synthetisiert wird. TFF1 und TFF2 sind nicht detektierbar. Im Gegensatz dazu lassen sich alle drei Peptide TFF1-3 in SF auf Protein-Ebene nachweisen. In Bezug auf die Quantifizierung der TFF-Peptide zwischen den einzelnen Gruppen zeigen sich für TFF1 keine signifikanten Veränderungen. TFF2 ist in SF von Patient*innen, die an RA leiden, im Verhältnis zu OA signifikant hochreguliert. Bei TFF3 zeigt sich eine signifikant niedrigere Proteinkonzentration bei OA im Verhältnis zu gesunder SF sowie eine signifikant höhere Konzentration bei RA im Verhältnis zu OA. Betrachtet man die Expression und Synthese von TFF3 in SM, sind keine signifikanten Unterschiede zwischen RA, OA und gesunden Proben offensichtlich. Schlussfolgerungen: Die Untersuchungen belegen, dass ausschließlich TFF3 durch die SM produziert wird. In der SF können jedoch unerwarteterweise alle drei TFF-Peptide nachgewiesen werden. Da weder Gelenkknorpel noch SM TFF1 und TFF2 produzieren und SM bekanntermaßen ein Ultrafiltrat des Blutserums ist, ist davon auszugehen, dass TFF1 und TFF2, sowie wahrscheinlich zusätzlich zur Eigenproduktion durch die SM auch TFF3, aus dem Blutserum in die SF diffundieren. Weitere Untersuchungen sind notwendig, um eine Aussage über eine mögliche Funktion der TFF-Peptide im Rahmen der Pathogenese der RA und der OA sowie ggf. mögliche weitere Therapieoptionen zu treffen. Insbesondere werden für eine Einschätzung der Quantifizierung der Peptide in den unterschiedlichen Gruppen mehr Patient*innen-/Personendaten benötigt. Relevant sind z.B. Begleiterkrankungen oder eingenommene Medikamente, die u.a. den Blutserumspiegel der TFF-Peptide beeinflussen können.
... [27] Baru-baru ini, sebuah protein baru dari film air mata milik keluarga protein surfaktan, palate lung nasal clone (PLUNC), telah ditemukan meningkat dalam air mata pasien DED. [28] Neuromediator, seperti zat P, calcitonin generelated peptide (CGRP), neuropeptide Y (NPY), vasointestinal peptide (VIP) dan neural growth factor (NGF), juga telah ditentukan dalam air mata pasien DED dan berkorelasi dengan temuan klinis. [29] Secara khusus, kadar NGF dalam air mata ditemukan meningkat secara signifikan pada pasien DED, sedangkan CGRP dan NPY menurun secara signifikan, dibandingkan dengan subjek sehat. ...
Pendahuluan: Diagnosis penyakit mata kering (Dry Eye Disease/ DED) terutama pada tahap awal merupakan hal yang penting, tetapi seringkali sulit. Hal ini dikarenakan kurangnya standar emas dan korelasi yang buruk antara perubahan biokimia air mata dan tanda-tanda klinis. Biomarker air mata dinilai dapat digunakan dalam diagnosis dan memantau DED karena bersifat non invasive, serta memiliki korelasi yang baik dengan perubahan biokimia air mata dan perkembangan penyakit. Tujuan: Artikel ini akan memaparkan beberapa biomarker air mata yang paling penting untuk DED yaitu marker untuk disfungsi kelenjar lakrimal, inflamasi, stres oksidatif, dan intoleransi lensa kontak, serta korelasinya dengan subtipe dan keparahan penyakit. Metode: Metode yang digunakan adalah studi pustaka dengan menggunakan jurnal 10 tahun terakhir yang diperoleh dari mesin pencarian seperti Sciencedirect, PubMed, Google Scholar dan ClinicalKey. Pembahasan: Biomarker untuk disfungsi kelenjar lakrimal ditandai dengan perubahan kadar protein (laktoferin, lisozim, dll), neuromediator (NGF, CGRP, NPY, Serotonin), dan mucin ((MUC)5AC); sementara respons inflamasi ditandai dengan perubahan ekspresi sitokin, kemokin, MMP-9, dan albumin. Stres oksidatif ditandai dengan perubahan kadar lipid (HNE, MDA). Sementara itu intoleransi lensa kontak dihubungkan dengan perubahan secretoglobin 1D1, β2 microglobulin, lacritin, secretoglobin 1A2, albumin, LPRR4, LCN-1, dan PIP. Kesimpulan: MMP-9 dan kombinasi Mammaglobin B, lipophilin A, dan B2MG merupakan biomarker dengan sensitivitas dan spesifitas tertinggi dari biomarker lainnya. Beberapa biomarker tersebut dapat digunakan untuk mendiagnosis DED, membedakan antara sindrom Sjögren DED dan sindrom non-Sjögren DED, ADDE dari EDE, serta menentukan tingkat keparahan penyakit.
... 26 Perilaku personal hygiene sebagai perwujudan perilaku hidup bersih dan sehat (PHBS) akan berdampak positif pada hasil yang akan didapatkan begitupun dengan aspek fisik dan psikis seorang peserta didik, karena kondisi tersebut berkontribusi terhadap aktivitas proses belajar seorang peserta didik Aktivitas pembelajaran yang matang dengan persiapan maksimal pada gilirannya akan berpengaruh besar terhadap prestasi belajar yang optimal. 28,29 ...
... Studi pendahuluan ini menunjukkan bahwa FAE mungkin aman dan menguntungkan pada pasien yang menderita necrobiosis lipoidica. [28] ...
... [23] Pasien dalam penelitian ETOMS yang diobati dengan interferon beta-1a rekombinan (Rebif) menunjukkan penurunan 24% dalam terjadinya MS secara klinis selama jangka waktu 2 tahun dibandingkan dengan pasien yang menerima plasebo. [28] Pasien yang dirawat menunjukkan perubahan pada MRI mereka seperti yang terlihat pada percobaan CHAMPS. [3,29] Penyakit ini dikarakteristikkan oleh rasa nyeri yang timbul akibat kuku yang tumbuh memotong salah satu atau kedua sisi paronikium. ...
Background: Health prevention and promotion are both important in making better public health. In order to actualize both aspects, Posyandu cadre play major role. Cadre have bigger chance and impact to educate the people who are living around them. However, doing a direct education has become more difficult since physical contacts were minimalized during this COVID-19 outbreak. Therefore, an effective tele-education is needed as an effort to prevent COVID-19 transmission. The study aims to evaluate the effect of tele-education through Youtube and Whatsapp to enhance people's understanding on COVID-19 transmission prevention Method: The study was a cross-sectional study with observational descriptive-analytical methods and quantitatively approach. Subject of the study was Posyandu cadre in Burangrang Village, Lengkong District, Bandung with the subject total was 19. Results: After given a tele-education, there was an increase in total score means between pretest and posttest with p=0.000 (p<0.001). Moreover, there were increases in both social media’s impression and engagement on educational video which was uploaded on Youtube. Discussion: In doing tele education to Posyandu cadre, video as a media to deliver the content was more preferred. Sharing the knowledge through video along with evaluating participants' understanding of the given topics could enhance cadre knowledge about handwashing as a prevention in the middle of COVID-19 outbreak. Conclusion: Tele education using video could enhance cadre’s understanding about handwashing. The media that has been used was able to deliver the content based on impression and engagement evaluations.
... Auch PLUNC zeigt in allen Immunfluoreszenzuntersuchungen der Tränendrüse ein positives Signal. Dies bestätigt die Ergebnisse von Schicht et al. (2015), die PLUNC als Sekretionsprodukt der Tränendrüse im Tränenfilm identifiziert haben. Da die Tränendrüse, gemeinsam mit den akzessorischen Tränendrüsen, Hauptproduktionsort der wässrigen Phase des Tränenfilms ist (Holly und Lemp, 1977), könnte der Flüssigkeitsgehalt des Tränenfilms durch Modulation der ENaC-Akitivität beeinflusst werden. ...
... Wie bei Schicht et al. (2015) kann PLUNC in der vorliegenden Arbeit im Epithel des Augenlids und in den Meibomdrüsen nachgewiesen werden. Ebenso werden erstmals alle ENaC-Untereinheiten in Kolokalisation mit PLUNC beschrieben. ...
... Dies bestätigt die Ergebnisse früherer Studien, im Rahmen derer PLUNC in Kornea ebenfalls in deutlich höherer Konzentration als in Konjunktiva nachgewiesen wurde . Ebenso beobachteten Schicht et al. (2015) in vorherigen Untersuchungen an denselben HCE-und HCjE- Außer der Elektrolyt-und Flüssigkeitshomöostase (Kellenberger und Schild, 2002), besitzt der ENaC auch Bedeutung im Rahmen der Wundheilung (Chifflet et al., 2005 (Hobbs et al., 2013). Garcia-Caballero et al. (2009) (Schild et al., 1997). ...
Das Protein palate lung nasal clone (PLUNC) ist Bestandteil der Augenoberfläche und des Tränensystems des Menschen und wird beim Trockenen Auge vermehrt in die Tränenflüssigkeit sezerniert. Durch Interaktion mit dem epithelialen Natriumkanal (ENaC, epithelial sodium channel, epithelial Na+ channel) kann PLUNC das Flüssigkeitsvolumen der Atemwegsoberfläche beeinflussen. Ziel dieser Arbeit ist der Nachweis der vier bekannten ENaC-Untereinheiten an der Augenoberfläche und im Tränensystem sowie deren Kolokalisation mit PLUNC. Ferner sollen der Einfluss von PLUNC und ENaC an der Augenoberfläche und eine mögliche Bedeutung im Hinblick auf das Trockene Auge untersucht werden. Polymerase-Kettenreaktion (PCR), Western Blot und Immunfluoreszenz- Doppelfärbungen dienen dem Nachweis von ENaC und PLUNC am Auge. Mittels enzyme-linked immunosorbent assay (ELISA) wird die PLUNC-Konzentration in humanen Kornea- und Konjunktivaepithelzellen (HCE, HCjE) nach Stimulation mit Amilorid untersucht. Mit dem electric cell-substrate impedance sensing (ECIS) System wird der Einfluss von rekombinantem humanen PLUNC Protein (rhuPLUNC) und Amilorid auf das Wachstum von HCE- und HCjE-Zellen nach Verwundung untersucht. Alle vier ENaC-Untereinheiten sind Bestandteil der Augenoberfläche und des Tränensystems und dort mit PLUNC kolokalisiert. Die Stimulation von HCE- und HCjE- Zellen mit Amilorid führt besonders in HCjE-Zellen zu einem signifikanten Anstieg der PLUNC-Konzentration im Vergleich zum Ausgangswert. Nach Verwundung von HCE- und HCjE-Zellen beeinflusst Amilorid die Wundheilung negativ. RhuPLUNC kompensiert diesen Effekt teilweise. ENaC und PLUNC sind Bestandteile des okulären Systems und beeinflussen in vitro die Wundheilung der Augenoberfläche. Die endogene PLUNC-Expression unterliegt vielfachen Einflüssen und zwischen PLUNC und ENaC besteht ein Interakti- onspotential. RhuPLUNC beeinflusst die Wundheilung der Augenoberfläche unter bestimmten Umständen positiv und sollte als mögliche Therapieoption in der Be- handlung des Trockenen Auges weiter analysiert werden.
... Each sample (30 µg protein) was mixed with reducing sample buffer (RSB) containing 0.5 M dithiothreitol (DTT), 50% glycerin, 0.05% bromphenolblau, 20% β-mercaptoethanol (β-ME) and boiled for 5 minutes. SDS-PAGE and Western blot analyses were performed as described by [56]. The membrane was incubated with primary antibodies to TFF1 (dilution 1:100), TFF2 (dilution 1:70), and TFF3 (dilution 1:400) at 4 • C overnight before applying the secondary antibodies (dilution 1:4000) conjugated to horseradish peroxidase for 2 h and detecting the bands by chemiluminescence using ECL Plus (Amersham Pharmacia, Uppsala, Sweden). ...
Objective:
Trefoil factor family peptide 3 (TFF3) has been shown to support catabolic functions in cases of osteoarthritis (OA). As in joint physiology and diseases such as OA, the synovial membrane (SM) of the joint capsule also plays a central role. We analyze the ability of SM to produce TFF compare healthy SM and its secretion product synovial fluid (SF) with SM and SF from patients suffering from OA or rheumatoid arthritis (RA).
Methods:
Real-time PCR and ELISA were used to measure the expression of TFFs in healthy SM and SM from patients suffering from OA or RA. For tissue localization, we investigated TFF1-3 in differently aged human SM of healthy donors by means of immunohistochemistry, real-time PCR and Western blot.
Results:
Only TFF3 but not TFF1 and -2 was expressed in SM from healthy donors as well as cases of OA or RA on protein and mRNA level. In contrast, all three TFFs were detected in all samples of SF on the protein level. No significant changes were observed for TFF1 at all. TFF2 was significantly upregulated in RA samples in comparison to OA samples. TFF3 protein was significantly downregulated in OA samples in comparison to healthy samples and cases of RA significantly upregulated compared to OA. In contrast, in SM TFF3 protein was not significantly regulated.
Conclusion:
The data demonstrate the production of TFF3 in SM. Unexpectedly, SF contains all three known TFF peptides. As neither articular cartilage nor SM produce TFF1 and TFF2, we speculate that these originate with high probability from blood serum.
... Schicht et al. detected higher levels of palate lung nasal clone in DED patients, especially ADDE patients. [46] Malate dehydrogenase 2 (MDH2) is the mitochondrial isoform of malate dehydrogenase, an essential enzyme in the citric acid cycle. It participates in intracellular signal transduction. ...
In recent years, there has been increasing scientific interest in the use of tear film biomarkers in the diagnosis and management of dry eye disease (DED), owing to their potential important roles in the pathogenesis of ocular surface damage, as well as the technical feasibility of tear sample collection techniques. An Entrez PubMed search was conducted on March 2, 2019, to include papers investigating the use of tear film biomarkers in DED, and the results were classified according to whether the DED is associated with systemic inflammatory disease or not and further classified within each section according to the molecular nature of the biomarker for further discussion. A total of 58 relevant articles were reviewed. Certain cytokines, including interleukin-6 (IL-6), tumor necrosis factor-alpha, IL-17, and IL-8, were found by a number of studies to consistently reflect disease severity well and had strong correlations with tear film metrics and tests for ocular surface damage in dry eye without systemic inflammatory disease. For dry eye with systemic inflammatory disease, IL-17, IL-8, and IL-1 receptor antagonists were shown to be consistently higher in affected eyes and correlated well with ocular surface disease severity in more than one type of inflammatory disease. With the advancement in technology and lowered costs in the future, tear film biomarker counts would allow better diagnosis and monitoring of DED, as well as facilitate personalized treatment strategies.
... Tissues. The preparation of the samples were performed as previously described by Schicht et al. 15 . All tissue samples were obtained from body donors (5 male, 11 female, aged 33-76 years) donated to the Department of Anatomy and Cell Biology, Martin Luther University Halle-Wittenberg, Germany. ...
... Tear fluid. Tear fluid extraction were performed as previously described by Schicht et al. 15 . Tear fluid samples were collected with Schirmer strips at the Department of Ophthalmology, Friedrich Alexander University Erlangen-Nürnberg, Germany. ...
... Cell culture. Cell culture experiments were performed as previously described by Schicht et al. 15 6 ) were seeded in Petri dishes and cultured until confluence was reached. Cells were washed with phosphate buffered saline (PBS) and changed to serum-free medium for 3 h. ...
The study aimed to characterize the expression and function of SFTA3 at the ocular surface and in tears. Ocular tissues, conjunctival (HCjE) and human corneal (HCE) epithelial cell lines as well as tearfilm of patients suffering from different forms of dry eye disease (DED) were analyzed by means of RT-PCR, western blot, immunohistochemistry, and ELISA. A possible role of recombinant SFTA3 in corneal wound healing was investigated performing in vitro scratch assays. Tear film regulatory properties were analyzed with the spinning drop method and the regulation of SFTA3 transcripts was studied in HCE and HCjE after incubation with proinflammatory cytokines as well as typical ocular pathogens by real-time RT-PCR and ELISA. The results reveal that human ocular tissue as well as tears of healthy volunteers express SFTA3 whereas tears from patients with DED showed significantly increased SFTA3 levels. In vitro wounding of HCE cell cultures that had been treated with recombinant SFTA3 demonstrated a significantly increased wound closure rate and rSFTA3 reduced the surface tension of tear fluid. The results indicate that SFTA3 at the ocular surface seemed to be involved in wound healing and the reduction of surface tension.
... 15 PLUNC1 is also present in the tear film. 16 Up to now, SPs were detected in many locations throughout the human body such as in glandular structures of the sinonasal tract, 17,18 airway submucosal gland cells, 19 lacrimal gland, 20 parotid and submandibular gland, 21 furthermore in skin, 22 brain, 23 testis, 24 female reproductive tract, 25 gastrointestinal tract, 26 and urinary tract. 27 Even in bacteria such as Staphylococcus aureus and P. aeruginosa, SPs and genes were proved with antibodies and primers to human SP-A, SP-B, SP-C, and SP-D. ...
Surfactant proteins in different glandular structures of the oral cavity display antimicrobial activity for protection of invading microorganisms. Moreover, they are involved in lowering liquid tension in fluids and facilitate secretion flows. Numerous investigations for studying the occurrence of surfactant proteins in glandular tissues were performed using different methods. In the oral cavity, minor salivary glands secrete saliva continuously for the maintenance of a healthy oral environment. For the first time, we could show that infantile labial glands show expression of the surfactant proteins (SP) SP-A, SP-B, SP-C, and SP-D in acinar cells and the duct system in different intensities. The stratified squamous epithelium of the oral mucosa revealed positive staining for SPs in various cell layers.
