9,565 research outputs found
LL-37 induced HA metabolism and inflammatory cytokines and mediators.
No full text(A) SW982 cells were treated with varied concentrations of LL-37 for 72 hr. Culture media were collected and measured for HA concentrations. (B-H) SW982 cells were treated with 12.5 ng/mL of LL-37 for varied times. Cells were then collected and measured for the expression of TNF, IL1B, IL6, CXCL8, IL17A, CASP1, and PTGS2 genes, respectively. The values from triplicate experiments are expressed as the mean ± SD compared to the untreated control. Horizontal brackets indicate significance levels at *p < 0.05 or **p < 0.01.</p
LL-37 induced HA metabolism in SW982 cell line.
No full textSW982 cells were treated with 12.5 ng/mL of LL-37 for varied times. Cells were then collected and measured for the expression of HAS2, HAS3, HYAL1, HYAL2 and CD44 genes, respectively (A-E). The values from triplicate experiments are expressed as the mean ± SD of fold increases compared to the untreated control. Horizontal brackets indicate significance levels at *p < 0.05 or **p < 0.01.</p
LL-37 induces activation of caspase-1 during airway epithelial cell infection with <i>P</i>. <i>aeruginosa</i>.
No full textA) FLICA Caspase 1 activation assay quantitation in NHBE cells, assessed by fluorescence microscopy, 3 hours post-treatment with either media only (control), 20 μg/ml LL-37, PAO1 at 10:1 MOI, PAO1 + LL-37, 10 ng/ml LPS for 3 hours, 5 mM ATP only for 1 hour, or 10 ng/ml LPS for 3 hours followed by 5 mM ATP for 1 hour +/- 20 μg/ml LL-37. Data represent means +/- SEM from n = 3–6 independent experimental repeats, *** p < 0.001, ** p < 0.01, by 2-way ANOVA with Bonferroni Post-test. B) Quantitation of percentage of cell death at 6 hours post-treatment, assayed by TUNEL staining in 16HBE14o- cells treated with either media only (control), 20 μg/ml LL-37, PAO1 at 10:1 MOI, or PAO1 + LL-37 +/- 25 μM YVAD-CHO Caspase 1 inhibitor. Data represent means +/- SEM from n = 5 independent experimental repeats, *** p < 0.001, ** p < 0.01, * p < 0.05, versus PAO1+LL-37 condition, by 2-Way ANOVA with Bonferroni Post-test, or by unpaired t-test for the separate evaluation of caspase-1 inhibition. C) Quantitation of apoptosis as measured by Caspase 3/7 Magic Red probe in NHBE cells, 3 hours post-treatment with either media only (control), 1 μM staurosporine, 20 μg/ml LL-37, PAO1 at 10:1 MOI, or PAO1 + LL-37. Data represent means +/- SEM from n = 3 independent experimental repeats, *** p < 0.001, ** p<0.01, * p<0.05 by 2-way ANOVA with Bonferroni Post-test, or by unpaired t-test to evaluate the positive control staurosporine. D) Confocal microscopy images, at high and low power magnification, of NHBE cells treated with 20 μg/ml LL-37 and PAO1 at 10:1 MOI, co-stained with Caspase 1 FLICA probe (green) and Caspase 3/7 Magic Red probe (red), showing mutual exclusivity of cell staining with these probes. Representative of n = 4 separate experiments.</p
LL-37 promotes expression of IL-1β by human and murine myeloid cells in a P2X7R-independent manner.
No full textA and B) Primary human blood-derived monocytes were isolated, pretreated with 10 ng/ml E. coli 0111:B4 LPS for 3 hours, then exposed for 1 hour to 50 μg/ml LL-37, 5 mM ATP or untreated. Supernatants were assessed for IL-1β by ELISA (A), comparing the effect of LL-37 or ATP on LPS-stimulated or unstimulated cells using 2-Way ANOVAs with Bonferroni post-tests (*** p<0.001, ** p<0.01, * p<0.05, data represent mean +/- SEM for n = 8 per condition), or by western blot (B) detection of pro- and mature IL-1β. (C) Primary human blood-derived monocytes pre-treated with 10 ng/ml LPS as in (A) and also treated with 1 μM KN62 prior to addition of 50 μg/ml LL-37 or 5 mM ATP. Supernatants were assessed for IL-1β by ELISA, and the impact of KN-62 was determined using unpaired t-test (*** p<0.001, data represent mean +/- SEM for n = 4 per condition). D and E) Murine peritoneal macrophages from WT or P2X7R-/- mice were exposed to LL-37 (10 or 50 μg/ml) or ATP (5 mM) for 30 minutes with supernatants analysed by ELISA, and assessed by 1-way ANOVA with Bonferroni multiple comparisons post-test, ** p<0.01, * p<0.05, data represent mean +/- SEM for n ≥ 3 per condition. F and G) Human MDM were treated with either 20 μg/ml LL-37, PAO1 at 10:1 MOI or 10 ng/ml LPS for 3 hours, or 5 mM ATP only for 1 hour, or 10 ng/ml LPS for 3 hours followed by 5 mM ATP for 1 hour +/- 20 μg/ml LL-37. Data represent mean +/- SEM for n = 3–6 per condition, *** p < 0.001, ** p<0.01, * p<0.05, by 2-way ANOVA with Bonferroni Post-test, with supernatants assessed for IL-1β by ELISA (F) and caspase-1 activity assessed by FLICA activity assay (G). ns = no significant difference. H) Real Time PCR detection of inflammasome components using commercial TaqMan Gene Expression assays for NLRC4, NLRP3, caspase 1 (Casp1) and caspase 4 (Casp4) in MDM (white columns) and NHBE (black columns) cells. ND = not detectable.</p
FREE FATTY-ACID AND DIACYLGLYCEROL ACCUMULATION IN THE RAT-BRAIN DURING RECURRENT SEIZURES IS RELATED TO CORTICAL OXYGENATION
No full textCerebral blood flow and oxygenation increase during the early seizures of a series, but the increase in cerebral blood flow attenuates during late seizures, sometimes resulting in decreased cortical oxygenation. Cortical free fatty acids (FFA) and diacylglycerols also increase during early seizures and the increase attenuates during late seizures. We analyzed the correlation between lipid accumulation and cortical O2 during periodic pentylenetetrazol-induced seizures. During early seizures, both FFA and diacylglycerols increased in the cerebral cortex, particularly arachidonate (20:4) and stearate (18:0). Changes in lipids were different during late seizures, depending on cortical O2 levels. An increase in cortical O2 during late seizures was associated with lower FFA levels compared with early seizures, and FFA levels recovered to basal levels during interictal periods. A decline in cortical O2 was associated with a further increase in FFA, which remained elevated during interictal periods. Our results indicate that periseizure lipid accumulation is related to cortical oxygenation
Hepatocellular carcinoma associated with focal nodular hyperplasia: report of a case with clonal analysis.
No full textThe antimicrobial peptide LL-37 in the oral cavity
No full textThe antimicrobial peptide LL-37 is an important molecule of innate immunity in the oral cavity. Aim of the doctorate study was to examine the concentration of LL-37 in saliva and its possible correlation to various factors. The whole study group consisted of 203 subjects with good general health: 49 children (1-18 years old), 76 adult subjects with good periodontal health (19-78 years old), 58 subjects with chronic periodontitis (19-83 years old), 20 edentulous subjects (48-85 years old). Twelve patients with oral lichen planus (46-74 years old) were also examined. The salivary concentration of LL-37 (in ng/ml) was determined by an enzyme-linked immunoabsorbent assay (ELISA). The peptide was detected in all saliva samples. Its concentration considerably varied in all groups studied, besides the totally edentulous subjects. The median value of salivary LL-37 in children was 22 ng/ml, and a positive significant correlation was observed between peptide concentration and age. Also, children with high caries activity exhibited significantly lower peptide salivary levels than caries free children and children with low to moderate caries activity. Salivary levels of LL-37 did not differ between the healthy adult group (30.5 ng/ml, range 0.75-285) and the group of subjects with chronic periodontitis (22.52 ng/ml, range 1-207). Edentulous subjects (1.81 ng/ml, range 0.15-4.4) demonstrated significantly lower peptide levels than the healthy and the chronic periodontitis group. Subjects with oral lichen planus exhibited higher concentration of LL-37 (median value 54.25, range 10.5-760) than age-matched healthy subjects (33 ng/ml, range 0.75-166) but the difference was on the verge of statistical significance. The doctorate thesis adds new knowledge regarding the presence of the antimicrobial peptide LL-37 in saliva and its role in the oral cavity, enhancing the prospect of possible future use as a theurapeutic molecule or a prognostic tool.Το αντιμικροβιακό πεπτίδιο LL-37 αποτελεί σημαντικό μόριο της φυσικής ανοσίας στο στοματικό περιβάλλον. Σκοπός της διδακτορικής διατριβής ήταν ο προσδιορισμός της σιελικής συγκέντρωσης του αντιμικροβιακού πεπτιδίου LL-37 καθώς και η διερεύνηση παραγόντων που πιθανώς σχετίζονται με αυτήν. Για το λόγο αυτό, έγινε λήψη ολικού σάλιου από 203 άτομα, με καλή γενική υγεία 49 ανήλικα υγιή άτομα (1-18 ετών), 76 ενήλικα υγιή άτομα (19-78 ετών), 58 άτομα με χρόνια περιοδοντίτιδα (19-83 ετών), 20 νωδά άτομα (48-85 ετών). Εξετάσθηκαν επίσης 12 ασθενείς με στοματικό ομαλό λειχήνα (46-74 ετών). Η συγκέντρωση του πεπτιδίου στο σάλιο (σε ng/ml) προσδιορίστηκε με την ανοσοενζυμική μέθοδο ELISA. Το πεπτίδιο LL-37 ανιχνεύθηκε σε όλα τα δείγματα σάλιου. Η συγκέντρωση του παρουσίαζε μεγάλη διακύμανση εκτός από την ομάδα των νωδών ατόμων. Στην ομάδα των ανήλικων ατόμων παρατηρήσαμε θετική συσχέτιση με την ηλικία και τον τύπο της οδοντοφυΐας. Επιπλέον, τα παιδιά με υψηλή τερηδονική δραστηριότητα είχαν σημαντικά χαμηλότερα επίπεδα του πεπτιδίου σε σχέση με τα παιδιά που ήταν ελεύθερα τερηδόνας και τα παιδιά με χαμηλή προς μέτρια τερηδονική δραστηριότητα. Τα επίπεδα του πεπτιδίου στο σάλιο δεν διέφεραν μεταξύ της ομάδας των υγιών ενηλίκων (διάμεση τιμή 30,5 ng/ml, εύρος 0,75-285) και της ομάδας των ασθενών με χρόνια περιοδοντίτιδα (διάμεση τιμή 22,52 ng/ml, εύρος 1-207). Τα νωδά άτομα (διάμεση τιμή 1,81 ng/ml, εύρος 0,15-4,4) παρουσίαζαν σημαντικά χαμηλότερα επίπεδα του LL-37 σε σχέση με την ομάδα των υγιών ατόμων και των ατόμων με χρόνια περιοδοντίτιδα. Η ομάδα των ατόμων με ομαλό λειχήνα παρουσίαζε υψηλότερες συγκεντρώσεις του LL-37 (διάμεση τιμή 54,25 ng/ml, εύρος 10,5-760) σε σχέση με αντίστοιχης ηλικίας άτομα χωρίς ομαλό λειχήνα (διάμεση τιμή 33 ng/ml, εύρος 0,75-166), παρόλο που η διαφορά δεν ήταν στατιστικά σημαντική. Τα παραπάνω προσθέτουν νέα γνώση σχετικά με την παρουσία του πεπτιδίου LL-37 στο σάλιο και το ρόλο του στη στοματική κοιλότητα, ενισχύοντας την προοπτική σχετικά με πιθανές μελλοντικές χρήσεις του ως θεραπευτικού παράγοντα ή δείκτη φλεγμονής
Vitamin D and LL-37 in Serum and Saliva: Insights into Oral Immunity
No full text(1) Background: In recent years, there has been a growing interest in understanding the innate immunity of the mouth, particularly the mechanisms through which vitamin D influences oral health. Researchers have increasingly focused on the association between vitamin D and the antimicrobial peptide LL-37 since the CAMP gene, responsible for encoding the LL-37 peptide, is a direct target of both vitamin D and its receptor (vitamin D receptor, VDR). This study aimed to explore the correlation between the 25-hydroxyvitamin D (25(OH)D) levels and the concentration of the LL-37 peptide in both serum and saliva. The objective was to compare the serum concentrations of 25(OH)D and ll-37 with those in saliva and to access the correlations between the two compounds. (2) Methods: Serum and whole saliva samples were collected from 72 healthy adults (mean age 28.68 ± 8.35). The levels of 25(OH)D and LL-37 were assessed in both the saliva and serum samples using commercially available enzyme-linked immunosorbent assay (ELISA) kits. (3) Results: The 25(OH)D levels in the serum (median 5.92 ng/mL, min–max 2.7–10.4 ng/mL) correlated with the LL-37 serum levels (62 ng/mL, min–max 18–378 ng/mL; Pearson’s r 0.328, p = 0.005). Additionally, the 25(OH)D levels in saliva (median 1.16 ng/mL, min–max 0.54–2.12 ng/mL) strongly correlated with the LL-37 salivary levels (median 44 ng/mL, min–max 6.5–205 ng/mL; Pearson’s r 0.667, p < 0.001). The 25(OH)D salivary levels demonstrated a robust correlation with the LL-37 salivary levels. (4) Conclusions: This discovery emphasizes the complex interplay between vitamin D and LL-37 and lay the groundwork for the further exploration of vitamin D’s role in oral immune function
Perbandingan Kadar Serum LL-37 Pada Pasien Morbus Hansen Tipe Multibasiler Dan Pausibasiler
No full textMorbus Hansen (MH)/penyakit kusta merupakan penyakit hingga saat ini
masih menjadi permasalahan kesehatan yang besar. Banyak faktor yang
mempersulit penegakan diagnosis, banyak kasus MH yang lolos pendeteksian
hingga penanganannya terlambat dan mengakibatkan disabilitas yang signifikan.
Spektrum klinis MH dipengaruhi oleh cell mediated immunity yang utamanya
diperankan oleh peptida antimikroba/antimicrobial peptides (AMP). LL-37
merupakan AMP yang berfungsi untuk membunuh bakteri serta berperan dalam
imunitas bawaan. Penelitian ini bertujuan untuk mengetahui perbedaan kadar LL-
37 serum pada pasien MH tipe MB dan PB di RSUD Dr. Saiful Anwar Malang dan
RS Kusta Kediri Desain studi yang digunakan adalah pendekatan potong lintang
dengan metode analisis komparatif. Sampel diambil menggunakan metode
consecutive sampling. Sampel darah vena diambil dari subyek kemudian
dilakukan pengukuran kadar serum LL-37 menggunakan ELISA kit. Didapatkan
rerata kadar serum LL-37 pada pasien MH tipe PB sebesar 94,15 ng/ml ± 79,92
dengan kadar terendah 43,429 dan tertinggi 260,268. Rerata kadar serum LL-37
pada pasien MH tipe MB sebesar 32,73 ng/ml ± 14,58 dengan kadar terendah
5,714 ng/ml dan kadar tertinggi 43,857 ng/ml. Hasil uji Mann Whitney kadar serum
LL-37 pada MH tipe MB dan PB menunjukkan nilai sig (0,000) < α(0.05). Terdapat
perbedaan yang bermakna secara statistik terhadap kadar serum LL-37 antara
pasien MH tipe MB dan PB (p = 0,000). Dapat disimpulkan bahwa kadar serum
LL-37 pasien MH tipe MB berbeda signifikan dengan kadar serum LL-37 pasien
MH tipe PB, dimana kadar serum LL-37 pasien MH tipe MB lebih rendah dibanding
tipe PB
Vitamin D and innate immunity in pneumonia and COPD
No full textA resurgence of interest in vitamin D research has led to the discovery that it plays a role in an unexpectedly large number of biological processes, and that reduced levels of this hormone are implicated in a range of diseases. In fact, it is estimated that vitamin D is involved in the regulation of 3% of the human genome. Two genes containing the target sequence indicative of vitamin D regulation are those encoding LL-37 and hBD-2. These antimicrobial peptides are integral components of the innate immune system and act as natural antibiotics to help combat infection.
The respiratory epithelium exposes a large surface area to environmental pathogens, making the innate immune response extremely important in its defence. Microbial infection of the respiratory tract is the cause of pneumonia, and is implicated in cases of COPD exacerbation. This study aimed to determine whether a relationship existed between vitamin D, LL-37 and hBD-2 in 185 patients admitted to Waikato hospital with either condition. It was hypothesised that low vitamin D would correlate with reduced peptide levels, and that this would be associated with increased infection severity and higher mortality rates.
Peptide concentrations in patient plasma were measured by indirect ELISA and compared to 25D levels. Statistical analysis revealed no significant associations between vitamin D status, peptide levels and severity, but did show increased mortality in individuals with severe vitamin D deficiency or low LL-37.
Based on the significance of LL-37 as a predictor of mortality (particularly in COPD), development of a plasma screening method using MALDI-TOF mass spectrometry was attempted, as a potential means of identifying patients most at risk. The success of this method was limited however, as the low abundance and small size of the mature peptide caused detection problems.
A protocol for assessing the vitamin D binding protein (DBP) genotype was developed, as it influences baseline 25D levels and response to supplementation. The association between low vitamin D and mortality suggests that supplementation could improve survival rates and, as the supplement dose required for effectiveness is genotype-dependent, this method could allow determination of the appropriate amount to administer to at-risk individuals
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