Identification of the splice variants of Recepteur d'Origine nantais (RON) in lung cancer cell lines

RON receptor tyrosine kinase is a transmembrane protein directly involved in suppression of inflammation and its aberrant expression linked to cancers and metastasis. Efforts to block deregulated RON signaling in tumors using small molecule kinase inhibitors or antibodies have been complicated by the presence of unknown number/types of isoforms of RON, which, despite being structurally similar, localize differently and mediate varied functions. Current study was designed to identify the splice variants of RON transcripts formed by skipping of sequences between exons 9 and 14 for better understanding of isoform specific RON signaling in cancers. PCR amplification and bi-directional sequencing of a 901 bp cDNA sequence located between exons 9 to 14 of RON from lung cancer cell lines revealed the presence of two splicing variants formed by skipping of exons 11 and 11–13. Each of these transcripts was found in more than one cell line. Expressed sequence tag (EST) database search indicated that the splicing variant lacking exons 11–13 was a novel one. Here we conclude that the splice variants of RON lacking exon 11 and exons 11–13 were detected in several lung cancer cell lines. Novel variant formed by skipping exons 11–13, the sequence of which code for transmembrane region, is predicted to code for a truncated isoform that may be secreted out. Tumors may antagonize the ligand dependent anti-inflammatory function of wild-type RON by secreting out the ligand binding isoforms

Association of vitamin B12 with pro-inflammatory cytokines and biochemical markers related to cardiometabolic risk in Saudi subjects

Background: This study aimed to examine the relationship between changes in systemic vitamin B12 concentrations with pro-inflammatory cytokines, anthropometric factors and biochemical markers of cardiometabolic risk in a Saudi population. Methods: A total of 364 subjects (224 children, age: 12.99 ± 2.73 (mean ± SD) years; BMI: 20.07 ± 4.92 kg/m2 and 140 adults, age: 41.87 ± 8.82 years; BMI: 31.65 ± 5.77 kg/m2) were studied. Fasting blood, anthropometric and biochemical data were collected. Serum cytokines were quantified using multiplex assay kits and B12 concentrations were measured using immunoassay analyzer. Results: Vitamin B12 was negatively associated with TNF-α (r = −0.14, p < 0.05), insulin (r = −0.230, p < 0.01) and HOMA-IR (r = −0.252, p < 0.01) in all subjects. In children, vitamin B12 was negatively associated with serum resistin (r = −0.160, p < 0.01), insulin (r = −0.248, p < 0.01), HOMA-IR (r = −0.261, p < 0.01). In adults, vitamin B12 was negatively associated with TNF-α (r = −0.242, p < 0.01) while positively associated with resistin (r = 0.248, p < 0.01). Serum resistin was the most significant predictor for circulating vitamin B12 in all subjects (r2 = −0.17, p < 0.05) and in children (r2 = −0.167, p < 0.01) while HDL-cholesterol was the predictor of B12 in adults (r2 = −0.78, p < 0.05). Conclusions: Serum vitamin B12 concentrations were associated with pro-inflammatory cytokines and biochemical markers of cardiometabolic risks in adults. Maintaining adequate vitamin B12 concentrations may lower inflammation-induced cardiometabolic risk in the Saudi adult population

Sphingolipid serum profiling in vitamin D deficient and dyslipidemic obese dimorphic adults

Recent studies on Saudi Arabians indicate a prevalence of dyslipidemia and vitamin D deficiency (25(OH)D) in both normal weight and obese subjects. In the present study the sphingolipid pattern was investigated in 23 normolipidemic normal weight (NW), 46 vitamin D deficient dyslipidemic normal weight (-vitDNW) and 60 vitamin D deficient dyslipidemic obese (-vitDO) men and women by HPTLC-primuline profiling and LC-MS analyses. Results indicate higher levels of total ceramide (Cer) and dihydroceramide (dhCers C18\u201322) and lower levels of total sphingomyelins (SMs) and dihydrosphingomyelin (dhSM) not only in -vitDO subjects compared to NW, but also in \u2013vitDNW individuals. A dependency on body mass index (BMI) was observed analyzing specific Cer acyl chains levels. Lower levels of C20 and 24 were observed in men and C24.2 in women, respectively. Furthermore, LC-MS analyses display dimorphic changes in NW, -vitDNW and \u2013vitDO subjects. In conclusion, LC-MS data identify the independency of the axis high Cers, dhCers and SMs from obesity per se. Furthermore, it indicates that long chains Cers levels are specific target of weight gain and that circulating Cer and SM levels are linked to sexual dimorphism status and can contribute to predict obese related co-morbidities in men and women

SNPs in FNDC5 (irisin) are associated with obesity and modulation of glucose and lipid metabolism in Saudi subjects

Background: Irisin is a recently identified myokine that plays an important role in preventing obesity and insulin resistance. We investigated whether the common FNDC5 (irisin precursor) gene variants influence susceptibility to obesity and type 2 diabetes (T2D) and verified the impact of FNDC5 gene variants on serum irisin levels, glucose and lipid metabolism in a Saudi population. Methods: Genomic DNA from 814 (394 T2DM and 414 controls) subjects were genotyped for the five common SNPs (rs3480A/G, rs1746661G/T, rs1298190A/G, rs726344A/G and rs1570569G/T) of the FNDC5 gene using the TaqMan genotyping assay. Biochemical parameters and hematic concentrations of irisin and insulin as well as anthropometric indices were collected. Results: Serum irisin levels were higher in T2DM patients compared to controls (p < 0.0001). Analyses of FNDC5 SNPs showed that: 1) The rs3480 GG associates with decreased risk of obesity (p = 0.005; odds ratio: 0.48) and lower body mass index (BMI) values (p = 0.03). In addition, GGAAG was identified as the protective haplotype against risk of obesity (p = 0.001; odds ratio: 0.23). 2) The rs1746661 G allele associates with higher triglyceride (TG) levels (p = 0.019). 3) The rs157069 TT genotype associates with higher fasting insulin (p = 0.029) and HOMA-IR (p = 0.002) as well as with lower circulating irisin levels (p = 0.016). Conclusions: SNPs in FNDC5 gene correlates with obesity and glucose-lipid metabolism possibly because they modulate the serum levels of irisin

Favorable Changes in Fasting Glucose in a 6-month Self-Monitored Lifestyle Modification Programme Inversely Affects Spexin Levels in Females with Prediabetes

Spexin (SPX) is a novel peptide thought to have a role in various metabolic regulations. Given its presumed body-weight regulatory functions, we aimed to determine whether lifestyle intervention programs on weight loss and fasting glucose (FG) improvement among people with impaired glucose regulation also alter levels of circulating SPX. A total of 160 Saudi adult males and females with prediabetes were randomly selected from a larger cohort (N = 294) who underwent a 6-month lifestyle modification program to improve their glycemic status. Participants were split into two groups based on differences in glucose levels post-intervention, with the first 50% (improved group) having the most significant reduction in FG. SPX was measured at baseline and after 6 months. Changes in SPX was significant only in the improved group [baseline: median (Q1\u2013Q3) of 164 pg/ml (136\u2013227) vs follow-up: 176 pg/ml (146\u2013285); p &lt; 0.01]. When stratified by sex, the significant increase was observed only in females [159 pg/ml (127\u2013252) vs 182.5 (152,369.1); p &lt; 0.01]. Furthermore, SPX levels showed a significant inverse association with FG (\u3b2 = 120.22, p = 0.003) even after adjustment with age and BMI, again only in females. Circulating SPX levels increase over time in people with prediabetes, particularly women who responded favorably in a 6-month lifestyle intervention program. Whether an unknown mechanism regulating the sexual disparity seen in SPX levels post-intervention exists should be further investigated using a larger sample size

Vanhemmuus - yksi elämän vaativimpia tehtäviä : opas 0-3 kk ikäisen lapsen vanhemmille

Vanhemmuuden käsite ja odotukset ovat muuttuneet viimeisen sadan vuoden aikana. Painetta vanhemmuudelle luovat niin yhteiskuntapolitiikka kuin työelämäkin. Vanhemmat ovat entistä enemmän huolissaan omasta kelpoisuudestaan vanhempana. Myös huoli omasta jaksamisesta vanhempana on yleistä. Toimeentulo, työn vaatimukset ja vanhempien oma terveys huolestuttavat ja kuormittavat nykyvanhempia. Suuri osa vanhemmista tarvitsee tukea vanhemmuuteensa. Tukea kaivataan muun muassa konkreettisiin asioihin kuten lapsen kasvatukseen ja perushoitoon. Tämän opinnäytetyön tavoitteena oli tukea vanhempia heidän vanhemmuudessaan ja arjessaan lapsen kanssa. Opinnäytetyön tarkoituksena oli luoda terveyskasvatusmateriaali verkkoon Naantalin terveyskeskukselle. Terveyskasvatusmateriaali on tarkoitettu tueksi vastasyntyneen vauvan vanhemmille sekä tiedon lähteeksi. Materiaali julkaistaan verkko-oppaana, jolloin se on helposti saatavilla ja päivitettävissä. Oppaassa on kustakin aiheesta pieni alustus sekä linkkejä, jotka johdattavat vanhemmat lisätiedon äärelle. Opinnäytetyömme on menetelmältään toiminnallinen ja se on toteutettu yhteistyössä Naantalin terveyskeskuksen kanssa. Opinnäytetyö kuuluu Turun ammattikorkeakoulun ”Terveesti tulevaisuuteen” – hankkeeseen, joka on osa Turun ammattikorkeakoulun lakisääteistä tutkimus-, kehitys- ja innovaatiotoimintaa (TKI). Opinnäytetyömme toteutettiin kahdessa osassa. Ensimmäisen osan tuotos eli verkko-opas pohjautuu kirjallisuuskatsaukseen sekä Naantalin terveyskeskuksen lastenneuvolan terveydenhoitajien toiveisiin. Tarkoituksena on ollut tuottaa luotettavaa ja ajankohtaista tietoa vastasyntyneen vauvan tuomista iloista ja haasteista vanhempien tarpeet huomioiden. Kirjallisuuskatsauksessa on käsitelty vanhemmuutta ja sen tuomia haasteita. Erikseen on perehdytty siihen, millaista tukea äidit ja isät omilla tahoillaan kaipaavat. Lisäksi on paneuduttu siihen, miten neuvola ja yksittäinen terveydenhoitaja voivat tukea vanhempia näissä asioissa. Tavoitteena on, että terveydenhoitajat pystyvät hyödyntämään valmista opasta omassa ohjaustyössään vauvaperheiden parissa. Tavoitteena on, että vanhemmat saavat tukea vanhemmuuteensa sekä arjen apuvälineitä ja lisätietoa haluamistaan asioista luotettavaksi todettujen verkkosivujen kautta. Opinnäytetyön toisessa osassa tehtiin ammatillinen posteri mainostamaan verkko-opasta.The concept and expectations of parenthood have changed over the last hundred years. Parents are faced with pressures from society as well as working life. Even more than before, parents are concerned by their capabilities as parents, and coping as a parent is a common worry. Parents of today are concerned and burdened by issues of livelihood, work and health. Most parents need support with parenting. Among other things, support is needed in concrete matters such as child rearing and basic care. The object of this thesis was to support parents with parenting and with everyday life with their child. The purpose of the thesis was to create online health education material for Naantali Health Station. The intention of the health education material was to support parents of a newborn child and to provide information. The material will be released as an online guide which makes it easily available and easy to update. Each subject includes a brief introduction and links to additional information. The methods of our thesis are functional, and the thesis has been carried out in collaboration with Naantali Health Station. The thesis is part of the “Terveesti tulevaisuuten” project of Turku University of Applied Sciences. The project is part of the university’s statutory research, development and innovation operations (RDI). Our thesis was carried out in two parts. The product of the first part, i.e. the online guide, is based on the literature review and the wishes of the public health nurses of the Naantali child health clinic. The purpose has been to produce reliable and up-to-date information about the joys and challenges of having a newborn baby, taking the needs of parents into consideration. The literature review deals with parenthood and its challenges. Focus has been placed on what kind of support both mothers and fathers need seperately. In addition, we have examined how a child health clinic and an individual public health nurse can support parents in these matters. The aim is that public health nurses can utilise the finalised guide in their guidance work with families with infants. In addition, the guide is meant to help parents with parenting, and to provide every-day tools as well as reliable web information. In the second part of our thesis, a professional poster was made to advertise the online guide

Recepteur d'Origine nantais (RON) tyrosine kinase splicing variants lacking exons 18 and 19 occur ubiquitously in lung cancer

Background: Aberrant expression of RON, a MET family receptor tyrosine kinase, has been correlated to tumor growth and metastasis. Intense research efforts are on to target RON using small molecule tyrosine kinase inhibitors or specific antibodies. However, progress towards specific targeting of RON is hampered by a lack of understanding of the nature and number of isoforms of RON expressed by tumors. We hypothesize that formation of different isoforms via alternative splicing may be fundamental to the tumor promoting functions associated with aberrantly expressed RON in cancers. Methods: In this study, we analyzed the transcript sequence variations caused by alternative splicing in the C-terminal region of RON cDNA by PCR amplification and sequencing of five small cell lung carcinoma (SCLC) and seven non-small cell lung carcinoma (NSCLC) cell lines. Results: Results revealed the presence of two alternatively spliced variants, each caused by unique exon(s) deletion: a previously known transcript variant lacking exon 19 and a novel one lacking exons 18+19. The two alternatively spliced variants together with the wild-type transcript were detected in each of the 12 lung cancer cell lines analyzed. Combined loss of exons 18+19 results in an in-frame deletion of 303 nucleotides corresponding to 101 amino acids of the tyrosine kinase domain. Translation products of transcript variants lacking exons 18 and 19 are expected to dominant negatively inhibit ligand stimulated RON signaling. Conclusions: The ubiquitous presence of alternatively spliced transcripts and their translation products may affect quantitative expression analysis, either by immunological or PCR methods, by interfering with estimation of normal RON, leading to exaggerated values. Besides, RON isoforms with dominant negative activities may interfere with siRNA based functional analysis of wild-type RON

Novel splicing variants of recepteur d'origine nantais (RON) tyrosine kinase involving exons 15–19 in lung cancer

Background Altered expressions of receptor tyrosine kinases drive the growth and metastasis of several cancers. RON is a single pass transmembrane receptor tyrosine kinase (RTK) shown to be aberrantly expressed in various cancer types. However, target validation and successful therapeutic targeting of RON in cancers is hampered by the co-existence of unknown number/types of isoforms, which are structurally similar but functionally diverse. Objective The objective of this study was to identify differential splicing in the C-terminal region of RON transcripts to better understand RON signaling in cancers. mRNA transcript sequence between exons 14 and 20 of RON was PCR amplified and sequenced using cDNA from 10 SCLC and 13 NSCLC cell lines. Specific exon deletions were identified by aligning sequencing chromatograms with reference RON cDNA sequence. Results We identified the presence of four unique transcript sequence variants of RON formed through skipping of exons 15–19, 16–19, 16–17 and 16. The transcript variants, except the one lacking exons 15–19, were found in more than one cell line. Several cell lines contained two to four of these uniquely spliced transcript variants. dbEST (Expressed Sequence Tags database) or other DNA sequence databases did not contain RON cDNA sequences corresponding to any of the above exon deletions indicating that all these transcript sequence alterations are novel. Conclusions Results of our study indicate common occurrence of different types of alternatively spliced transcripts of RON in lung cancer with potential to be translated into proteins lacking active kinase domain. Our findings suggest that tumors produce several dominant negative isoforms which probably inhibit ligand dependent RON signaling, and hence, raise important questions regarding the appropriateness of blocking wild type RON signaling for therapy. Further, presence of transcript variants and their isoform products may interfere with quantitative and functional analysis during target validation. Abbreviations MST1R, macrophage stimulating 1 receptor; RTK, receptor tyrosine kinase; MSP, macrophage stimulating protein; SCLC, small cell lung cancer; NSCLC, non-small cell lung cancer; EGFR, epidermal growth factor recepto

Recepteur d'Origine nantais (RON) tyrosine kinase splicing variants lacking exons 18 and 19 occur ubiquitously in lung cancer

Aberrant expression of RON, a MET family receptor tyrosine kinase, has been correlated to tumor growth and metastasis. Intense research efforts are on to target RON using small molecule tyrosine kinase inhibitors or specific antibodies. However, progress towards specific targeting of RON is hampered by a lack of understanding of the nature and number of isoforms of RON expressed by tumors. We hypothesize that formation of different isoforms via alternative splicing may be fundamental to the tumor promoting functions associated with aberrantly expressed RON in cancers. Methods: In this study, we analyzed the transcript sequence variations caused by alternative splicing in the C-terminal region of RON cDNA by PCR amplification and sequencing of five small cell lung carcinoma (SCLC) and seven non-small cell lung carcinoma (NSCLC) cell lines. Results: Results revealed the presence of two alternatively spliced variants, each caused by unique exon(s) deletion: a previously known transcript variant lacking exon 19 and a novel one lacking exons 18+19. The two alternatively spliced variants together with the wild-type transcript were detected in each of the 12 lung cancer cell lines analyzed. Combined loss of exons 18+19 results in an in-frame deletion of 303 nucleotides corresponding to 101 amino acids of the tyrosine kinase domain. Translation products of transcript variants lacking exons 18 and 19 are expected to dominant negatively inhibit ligand stimulated RON signaling. Conclusions: The ubiquitous presence of alternatively spliced transcripts and their translation products may affect quantitative expression analysis, either by immunological or PCR methods, by interfering with estimation of normal RON, leading to exaggerated values. Besides, RON isoforms with dominant negative activities may interfere with siRNA based functional analysis of wild-type RON.N/

Bone metabolism markers are associated with neck circumference in adult Arab women.

Summary: The study aimed to determine whether neck circumference is associated with bone metabolism markers among adult Arab women and found modest but significant associations with bone resorption markers, suggesting that neck circumference, a surrogate measure of upper subcutaneous fat, influences bone turnover expression among adult females. Introduction: Body fat distribution is associated with decreased bone resorption and neck circumference (NC), a surrogate measure for upper body fat, has never been tested as a marker that can reflect bone turnover. This is the first study aimed to analyze the associations between NC and several bone biomarkers among adult Saudi women. Methods: This cross-sectional study included a total of 265 middle-aged Saudi women [86 non-obese (mean age 52.7 ± 8.1; mean BMI 26.9 ± 2.3) and 179 obese (mean age 50.6 ± 7.5; mean BMI 35.7 ± 4.5)] recruited from primary care centers in Riyadh, Saudi Arabia. Anthropometrics included BMI, NC, waist and hip circumferences, total body fat percentage (%), and blood pressure. Biochemical parameters included glucose and lipid profile which were measured routinely. Serum levels of 25(OH) D, parathyroid hormone, RANKl, sclerostin, C-terminal telopeptide of collagen I (CTX-I), Dkk1, IL1β, osteoprotegerin, osteopontin, and osteocalcin were measured using commercially available assays. Results: In all groups, NC was inversely associated with PTH (R = − 0.22; p < 0.05) and positively associated with osteoprotegerin (R=0.20; p < 0.05) even after adjustments for age and BMI. Using all anthropometric indices as independent variables showed that only NC explained the variance perceived in CTX-I (p = 0.049). In the non-obese, waist-hip ratio (WHR) was significantly associated with sclerostin (R = 0.40; p < 0.05) and body fat was significantly associated with osteopontin (R=0.42; p<0.05). Conclusion: NC is modestly but significantly associated with bone biomarkers, particularly the bone resorption markers, among adult Arab women. The present findings highlight the importance of NC as measure of upper body subcutaneous fat in influencing bone biomarker expression in adult females