Immunization of mice with plasmids coexpressing HPV16 E5 and the novel oncogenehWAPL

2008 ◽  
Vol 18 (3) ◽  
pp. 534-539
Author(s):  
C. X CAO ◽  
J. MA ◽  
M. XUN ◽  
X. XUE ◽  
P. CHEN ◽  
...  

The novel human oncogenehWAPLis associated with uterine cervical cancer. The HPV16 E5 oncoprotein could induce genomic instability in normal human cells. However, the mechanism of E5 interaction with hWAPL still awaits definition. In our present studies, the eukaryotic expression plasmids, pcDNA3-hWAPL and pcDNA3-hWAPL-E5 were constructed and carried out to vaccinate mice directly. The result that indicated the polyclonal antibody titer in immunized mice sera was increased by enzyme-linked immunosorbent assay. In addition, the proliferative responses of immunized mice spleen cells showed the optical densities values in vaccinated group remarkably higher than that in the control group. In conclusion, the recombinant plasmids could induce strong humoral and cellular immune response and exhibited great potential as therapeutic targets in the treatment of cervical cancer. However, the result didn't show significant difference in group with coexpression of HPV16 E5–hWAPL and group with only hWAPL expression. Consistent with these observations, we demonstrated that HPV16 E5 was not the optimal factor to cooperate with hWAPL in gene therapy.

2014 ◽  
Author(s):  
Γεώργιος Γεωργίου

Βackground: angiogenesis is seen during the multiple stages of carcinogenesis, aswell as during the process of surgical wound healing, a fact which has led tosubstantial debate over the last decades about the potential impact of surgery upon thefinal outcome of ceratin patients treated for breast cancer.Aim: the present research aims at investigating the potential effect of surgery on theprocess of angiogenesis, by studying a number of factors that are related to the latter,in patients suffering from breast cancer before and after the time of the procedure,whilst comparing these results with those of patients that were operated on their breastfor non-malignant disease.Material-Methods: blood from 10 female patients with breast adenocarcinoma(Study Group) was collected via venipuncture before surgery (labeled as PRO), aswell as on post-operative day 3 (labeled as D3) and day 7 (labeled as D7). Moreover,blood samples were also taken from 6 female patients with fibroadenoma (ControlGroup) before surgery (PRO) and on day 3 afetr surgery (D3). These samples weremeasured for detection of circulating levels of three established angiogenesisbiomarkers using ELISA (Enzyme-Linked ImmunoSorbent Assay): VascularEndothelial Growth Factor-A (VEFG-A), Interleukin-8 (IL-8) and basic FibroblastGrowth factor (bFGF or FGF-2). In addition, circulating transcripts of 84 agiogenesirelatedgenes were determined using RT-PCR (Real Time Polymerase ChainReaction). The two groups of patients were firstly compared to each other regardingtheir results. Also, patients belonging to the Study Group were analized at differenttime points regarding surgery. Finally, the results were investigated againstclinicopathological data and patient outcome.Results: using ELISA we were able to detect increased levels of circulating VEGF-Aand IL-8 in the Study Group patients compared to the Control Group patientspreoperatively (p=0,0381 and p=0,0218 respectively), while for bFGF there was nostatistically significant difference documented. Surgery resulted in a significantincrease in VEGF-A levels on D3 (p=0,0389) and D7 (p=0,0172) as compared toPRO levels. Perioperative kinetics of IL-8 showed a mild trend towards increase,which, however, was not statistically significant. Postoperative levels of bFGF wereslightly increased on D3, but on D7 they were even lower than preoperative values(p=0,0205). Using RT-PCR certain differences between the Study Group and theControl Group were recorded regarding the circulating transcripts of a great numberof angiogenesis-related genes preoperatively: upregulation of VEGF-C, EGF, IL-8,FGF-1, SPHK1, NRP1, LAMA5, COL4A3, TEK, EFNA3, EFNB2. AKT1, ITGB3,THBS1, CCL11, TIMP3 and downregulation of CXCL10. Moreover, mastectomyinduced an altered expression in several key-genes in breast cancer patients:upregulation of THBS1, COL4A3, BAI1, ITGB3 and downregulation of EREG,SERPIFN1, CXCL9, CXCL10, IL1B, CCL2, CXCL1, HIF1A, NOTCH4. Conclusions: patients suffering from breast cancer have a different angiogenic profilein comparison to patients with fibroadenoma, as documented through their differencesin circulating levels of angiogenic factors. These levels are greatly changed after thesurgical procedure. VEGF showed a transient increase, while bFGF initially increasedbut only to finally decrease to levels that were even lower than the preoperative ones.Moreover, mastectomy promoted a shift in the expression pattern of a broad panel ofangiogenesis-related gene transcripts.


Author(s):  
Nur Anna C Sa’dyah ◽  
Agung Putra ◽  
Bayu Tirta Dirja ◽  
Nurul Hidayah ◽  
Salma Yasmine Azzahara ◽  
...  

Introduction<br />Liver fibrosis (LF) results from the unregulated chronic wound healing process in liver tissue. Transforming growth factor-beta (TGF-β) is the major contributing cytokine of LF promotion through activation of quiescent hepatic stellate cells (HSCs) into myofibroblasts (MFs) and increased extracellular matrix (ECM) deposition such as collagen leading to scar tissue development. Mesenchymal stem cells (MSCs) have an immunomodulatory capability that could be used as a new treatment for repairing and regenerating LF through suppression of TGF-β. This study aimed to examine the role of MSCs in liver fibrosis animal models through suppression of TGF-β levels without scar formation particularly in the proliferation phase.<br /><br />Methods<br />In this study, a completely randomized design was used with sample size of 24. Male Sprague Dawley rats were injected intraperitoneally (IP) with carbon tetrachloride (CCl4), twice weekly, for eight weeks to induce LF. Rats were randomly assigned to four groups: negative control, CCl4 group, and CCL4 + MSC-treated groups T1 and T2, at doses of 1 x 106 and 2x106 cells, respectively. TGF-β levels were analyzed by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA and a least significant difference (LSD) was used to analyse the data. <br /><br />Results<br />The TGF levels of LF rat models decreased on day 7 after MSC administration. The levels of TGF-β in both MSC groups T1 and T2 decreased significantly compared with the control group (p&lt;0.05). The TGF-β suppression capability of T2 was optimal and more significant than that of T1.<br /><br />Conclusion<br />MSCs can suppress TGF levels in liver fibrosis induced rats.


2021 ◽  
Vol 67 (3) ◽  
pp. 1-4
Author(s):  
Shler Ghafoor Raheem

The inflammatory reaction is one of the complications in patients with coronary atherosclerosis. This study aimed to determine the diagnostic value of platelet-activating factor (PAF) compared with high sensitivity C reactive protein (hs-CRP) in coronary atherosclerotic patients. Fifty patients with coronary atherosclerosis and 30 subjects with normal angiography were considered as the control group attending Cardiac Center-Surgical Specialty Hospital - in Erbil city / Iraq. The levels of PAF and hs-CRP were estimated quantitatively using a sandwich enzyme-linked immunosorbent assay and a particle-enhanced immune turbid metric assay, respectively. Lipid profiles and some hematological indexes were also used in this study. The levels of the inflammatory biomarkers of PAF and hs-CRP increased significantly in the patients group compared with controls (p<0.05). Although the patients group showed the highest level of low-density lipoprotein (LDL), the difference was not significant (p>0.05) compared with the healthy control. However, the incidence of risk factors such as smoking and obesity showed a significant difference (p<0.05) in the patients group. Additionally, the PAF level correlated positively and significantly with hs-CRP (p<0.05), and negatively with high-density lipoprotein (HDL) (p>0.05). Although hs-CRP was a valuable diagnostic marker for coronary atherosclerosis, the PAF level showed to be a better prognostic indicator than hs-CRP in coronary atherosclerosis patients.


2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S232-S232
Author(s):  
H Korkmaz ◽  
K Fidan

Abstract Background In this study, we investigated the importance of netrin-1 levels in ulcerative colitis (UC) in clinical activity of the disease, and its association with other proinflammatory cytokines IL-6 and TNF-α. Methods This study is a type of case–control study. Sixty-seven patients with UC (36 of them activation, 31 of remission) and 50 healthy controls were included in the study. UC patients; ‘Truelove Witts clinical activity index by remission (n = 31), mild activation (n = 21), moderate activation (n = 6) and severe activation (n = 9) were divided into groups. Netrin, IL-6 and TNF-α measurements in plasma samples were performed using enzyme-linked immunosorbent assay kit. Results Between the patient group and the control group; there was a statistically significant difference between netrin-1, IL-6, TNF-α, neutrophil, platelet (p &lt; 0.05 for all). The plasma netrin-1 mean of UC with severe activation group (139.21 ± 48.09 pg/ml) was statistically significantly higher than that of the mild activation (p = 0,037), remission group (p = 0,001) and control group(p = 0,011). The plasma netrin-1 mean of UC with moderate activation group was statistically significantly higher than that of the mild activation(p = 0,045) and remission group(p = 0,004). Conclusion Our results reveal that plasma netrin-1 levels have been shown to be associated with UC activation, similar to proinflammatory cytokines such as TNF-α and IL-6, in UC.


2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Duygu Alkan ◽  
Berrak Guven ◽  
Cigdem Coskun Turer ◽  
Umut Balli ◽  
Murat Can

Abstract Background The purpose of this study was to investigate gingival crevicular fluid (GCF) and serum folate-receptor 1 (FOLR1) levels in subjects with different periodontal status. Methods The study consists of three groups: Healthy group (n = 15), gingivitis group (n = 15) and chronic periodontitis group (n = 15). Clinical periodontal parameters including probing pocket depth (PPD), clinical attachment level (CAL), gingival index (GI) and bleeding on probing (BOP) were assessed. GCF and serum samples were collected from each patient and were analyzed FOLR1 levels by enzyme-linked immunosorbent assay. Results The values of FOLR1 in GCF were higher in gingivitis and periodontitis groups than among patient in control group (p < 0.016). Serum FOLR1 levels showed no significant difference between the groups. A significant correlation was observed between FOLR1 levels of GCF and BOP (p < 0.05). Conclusions Our preliminary data suggest that FOLR1 is not useful in monitoring the periodontal disease. Further studies are necessary to clarify the role, regulation and function of folate and it’s receptors in the pathogenesis of periodontal disease.


QJM ◽  
2020 ◽  
Vol 113 (Supplement_1) ◽  
Author(s):  
M E Nasser ◽  
S M Shawky ◽  
E A M Sanad

Abstract Background The biology of claudins is a rapidly evolving field, and many intriguing questions remain unanswered. Although it had been assumed that the reason there are ≥24 isoforms of claudin is that each one has distinct permeability properties. The nephron displays a wide spectrum of claudins, whose distribution varies in each tubular segment. In diabetic nephropathy and glomerulonephritis the gene expression of claudin-1, is markedly upregulated in the podocyte, accompanied by a tighter filtration slit diaphragm (cell-cell junction made by the glomerular podocytes) and the appearance of TJ-like structures between the foot processes causing further podocytes injury and proteinuria. Aim of the work to assess urinary claudin -1 level as a marker of podocyte injury in patients with proteinuria. Patients and Methods it is a case control study which was conducted upon 90 subjects who were divided into three groups: group I included 30 patients with type II DM, group II included 30 patients with glomerulonephritis and group III had 30 healthy subjects as controls. Urinary claudin-1 level was measured by Enzyme linked Immunosorbent Assay (ELISA) Results In this study, we found that urinary claudin-1 level was significantly higher in diabetics group and GN group than in control group. In comparison between GN group and diabetic group, we found that urinary claudin-1 level was higher in GN group than in diabetics group but with no statistically significant difference between the two groups. Conclusion urinary claudin-1 level was significantly higher in diabetics and GN group and has positive correlation with uACR. So it can be used as marker of podocytes injury and proteinuria


2019 ◽  
Vol 32 (9) ◽  
pp. 1005-1011 ◽  
Author(s):  
Ali Heidarianpour ◽  
Elnaz Shokri ◽  
Tayebe Baghian ◽  
Behnaz Shokri

Abstract Background The aim of this study was to investigate the effect of 12 weeks of aerobic training, 4 weeks of detraining and use gonadotropin-releasing hormone agonist (GnRHa) on serum C-reactive protein (CRP) and cortisol levels in girls with central precocious puberty (CPP). Methods Forty-five girls (aged 6–8 years) with precocious puberty were randomly divided into three groups (medication, training and medicine + training groups). Fifteen healthy girls (without precocious puberty) were also included as the control group. Serum CRP and cortisol levels were measured at baseline by the enzyme-linked immunosorbent assay (ELISA) technique. Then, the experimental groups performed an aerobic training program for 3 days/week 20–75 min per day at 45–75% maximum heart rate for 12 weeks. The medication groups also received GnRHa during the study, once a month (1 mL every 4 weeks) by intramuscular injection. Serum CRP and cortisol levels were measured again 48 h after the last training session and also after 4 weeks of detraining. Results Analysis of variance (ANOVA) with repeated measures showed a significant decrease in CRP (p = 0.02) and cortisol levels (p = 0.01) in the training group and the medicine + training group. Detraining led to return of CRP and cortisol levels to the pre-training levels (p = 0.001). No significant difference in serum CRP (p = 0.43) and cortisol levels (p = 0.06) was observed in the medication group. Further, no significant difference was observed between groups in CRP and cortisol. Conclusions Long-term regular moderate training decreases inflammation indices, and detraining eliminates the benefits of training in girls with precocious puberty.


2017 ◽  
Vol 11 (03) ◽  
pp. 340-344 ◽  
Author(s):  
Piero Papi ◽  
Stefano Di Carlo ◽  
Daniele Rosella ◽  
Francesca De Angelis ◽  
Mario Capogreco ◽  
...  

ABSTRACT Objective: The aim of this case–control study was to compare patients with a healthy peri-implant environment and patients affected by peri-implantitis, evaluating the occurrence of antibodies to extracellular matrix (ECM) molecules. The authors hypothesized the presence of ECM autoantibodies in serum of peri-implantitis patients. Materials and Methods: Patients were divided into two groups: one with dental implants with a diagnosis of peri-implantitis and one control group with implants classified as being “healthy.” Enzyme-linked immunosorbent assay was performed on patients' sera to detect human antibodies to type I, III, IV, and V collagens, laminin, and fibronectin. Fisher exact test was performed to evaluate statistical association, with a significant P < 0.05. Results: Forty-two patients were enrolled in this study, 27 females (64.28%) and 15 males (35.72%) with a mean age of 53 ± 29.69 years (age range 32–74). The presence of antibodies to CIII was recorded in 6/21 (28.57%) patients of test group, compared to just 2/21 (9.52%) for the control group, showing a statistically significant difference (P < 0.05). Other antibodies tested were found to be not statistically significant or absent. Conclusions: Within the limitations of this study, it can be concluded that further studies, with larger sample and different design, are necessary to address the research purpose, evaluating possible associations between anti-ECM antibodies and peri-implantitis.


2003 ◽  
Vol 14 (7) ◽  
pp. 458-462 ◽  
Author(s):  
Maria Belazi ◽  
Alexandra Fleva ◽  
Drakoulis Drakoulakos ◽  
Despina Panayiotidou

Our objective was to investigate the concentrations of IgA and IgG antibodies to Saccharomyces cerevisiae in whole saliva and serum samples from HIV-infected patients and to compare them with the corresponding antibody values of healthy controls. A cross-sectional design was used. The test group consisted of 23 HIV-infected male individuals, aged 20-41 years old, free of any other systemic disease. Twenty healthy subjects aged 27-43 years old served as controls. Whole unstimulated saliva and blood were collected from all subjects. IgA concentrations in saliva and IgA and IgG concentrations in serum were measured by solid-phase enzyme-linked immunosorbent assay. Salivary antibody concentrations were calculated by reference to a pooled standard saliva obtained from 10 healthy males with high levels of anti- S. cerevisiae antibody activity. Total IgA and IgG concentrations were measured by nephelometry/tholocymetry assay. No significant difference was observed in salivary specific IgA and serum specific IgG levels to S. cerevisiae, while serum specific IgA were significantly lower in HIV infected patients compared to control group. Opportunistic infections due to S. cerevisiae, although rare, cannot be dismissed. This yeast can show a potential virulence in debilitated patients, therefore, further extensive investigation should be considered.


Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 2292-2292
Author(s):  
Patrick Van Dreden ◽  
Grigoris T Gerotziafas ◽  
Barry J Woodhams ◽  
Mourad Chaari ◽  
Robert Girot ◽  
...  

Abstract Abstract 2292 Introduction: The clinical course of sickle cell disease (SCD) is punctuated by episodic vascular occlusive events. The possibility that activation of the clotting system plays a contributory role in these complications is supported by abundant clinical data during both steady-state disease and pain crisis. Hydroxyurea therapy induces fetal haemoglobin, improves laboratory parameters and reduces acute clinical complications of SCD, but despite an abundance of evidence for coagulation and platelet activation, it remains incompletely defined whether these changes contribute to the reduced thrombin generation. This study is designed to evaluate coagulation profiles of patients with SCA in steady state and to determine whether hypercoagulable state is modified or not in patients on hydroxyurea therapy. Patients and Methods: We studied erythrocyte derived microparticles (Ed-MP) and platelet derived microparticles (Pd-MP) expressing or not expressing phosphatidylserine (PS) in patients with steady state SCD and we evaluated their specific procoagulant activity and their impact on thrombin generation process. A total of 92 steady state SCD patients were included in the study, of which 19 were under treatment with hydroyurea. The control group consisted of 30 healthy age and sex matched controls. Microparticles in whole blood were assessed using flow cytometry. Ed-MP and Pd-MP were identified using an anti-CD235 and CD41 monoclonal antibodies and annexin V. Thrombin generation in platelet poor plasma (PPL) was measured by CAT assay using PPP-reagent 5pM (Thrombinoscope, The Netherlands). Procoagulant phospholipid dependent activity in plasma was assessed by the Procoag-PPL assay (Diagnostica Stago, France). Thrombomodulin (TM) levels were measured by enzyme-linked immunosorbent assay (Elisa) Asserachrom thrombomodulin (Diagnostica Stago, Asnieres, France). Results: Hydroxyurea treated patients had lower counts of leukocytes, reticulocytes and platelets and an increased mean hemoglobin concentration as compared to non treated patients. Leukocyte and reticulocytes counts of treated patients were higher than those of controls. Platelets counts did not differ between treated and untreated patients. Patients on treatment with hydroxyurea had significantly lower levels of Ed-MP/PS+ and Ed-MP compared to untreated patients. The concentration of Pd-MP/PS+ and Pd-MP were not significantly different between hydroxyurea treated and non treated patients. The Ed-MP/PS+ showed a significant inverse correlation with Hb F (p<0.05). Thrombogram parameters, lag-time, ttPeak, Peak and MRI were significantly different between hydroxyurea treated patients and non treated patients. In hydroxyurea treated patients in contrast to the untreated ones no correlation was found between Ed-MP/PS+ and Ed-MP and parameters of thrombin generation. Among hydroxyurea treated patients 68% showed MRI levels higher than the UNL. Stratification groups of treated patients according to the levels of microparticles with Ed-MP/PS+ or Pd-MP/PS+ concentration higher than the UNL showed non significant difference compared to entirely group of patients. The PPL concentration was significantly lower in the SCD-treated patient compared to untreated patients (p<0.05). In contrast to platelet-derived-microparticles, the numbers of erythrocyte-derived-microparticles differed between patients with and without hydroxyurea during steady state. In patients treated with hydoxyurea, platelets were correlated with Ed-MP, Pd-MP with and without PS+ (p<0.05), but any of the others parameters showed one association. Procoagulant phospholipids and thrombomodulin were increased in SCD with and without hydroxyurea compared with controls group (p<0.05). Conclusion: Treatment with hydroxyurea result in decreases in plasma markers of thrombin generation, and may decrease coagulation activation by reducing PS expression on the surface of both RBCs and platelets in addition to being a NO donor hydro may also decrese haemostatic activation by its effect in decreasing the white blood cell count and particularly monocytes that express TF, furthermore the beneficial effects of hydroxyurea may be due to vasodilatationand decressed platelet and coagulation activation following NO production. Disclosures: Van Dreden: Diagnostica Stago: Employment. Gerotziafas:APHP: Employment. Woodhams:Diagnostica Stago: Employment. Chaari:APHP: Employment. Girot:APHP: Employment. Kartechi:APHP: Employment. Galea:APHP: Employment. Lionnet:APHP: Employment. Elalamy:APHP: Employment.


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