Double-antibody Sandwich Enzyme-linked Immunosorbent Assay Research Articles

Efficient Detection of Pre-Proinsulin by Double Antibody Sandwich ELISA

To detect residual pre-proinsulin (PPI) in recombinant human insulin production, an analytical method based on double-antibody sandwich ELISA was developed in this study. The BALB/c mice were immunized with PPI, and the hybridomas secreting anti-PPI monoclonal antibodies were obtained using the conventional cell fusion technique and ELISA screening. We purified the antibody using a Protein G gel column and identified its purity by SDS-PAGE. The sandwich ELISA was used to explore the pairing effect, and the specificity of the paired antibody was determined. We selected a paired antibody with relatively good specificity to establish sandwich ELISA, constructed a quantitative curve, and evaluated the accuracy and sensitivity of the method. Six anti-PPI monoclonal antibodies were obtained, named P1, P2, P3, P4, P5 and P6, of which P5 had the highest titer value. The sandwich ELISA method was established with P5 for plating and P2 as detection antibodies. The linear range of the quantitative curve of PPI by sandwich ELISA was 0.645 to 82.5 pg/mL, the recovery was 95% and the detection limit was 3.06 pg/mL. In this study, we prepared six anti-PPI monoclonal antibodies and established the sandwich ELISA method to detect PPI in process control and product release control for recombinant human insulin production.

New Weed Hosts for Tomato Brown Rugose Fruit Virus in Wild Mediterranean Vegetation.

Tomato brown rugose fruit virus (ToBRFV; genus, Tobamovirus, family, Virgaviridae) was first reported in 2015 infecting tomatoes grown under protected cropping in the Jordan Valley. Since then, ToBRFV has been detected in tomatoes grown in both protected and open fields across Jordan. The increased incidence of ToBRFV prompted this investigation of the potential role of natural weed hosts in the dissemination of ToBRFV. A survey was conducted in the Jordan Valley and highlands to determine possible reservoir hosts of ToBRFV in fields and greenhouse complexes in which tomatoes were grown. Detection of ToBRFV infection was made by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and further confirmation by reverse-transcription polymerase chain reaction (RT-PCR), followed by DNA cloning and sequencing, and bioassays. Thirty weed species belonging to twenty-six genera from sixteen families were tested. Twelve species belonging to eight families were infected of which ten species are newly reported hosts for ToBRFV. Seed transmission of ToBRFV in Solanum nigrum was confirmed in a grow-out experiment. To our knowledge, this is the first report of the natural occurrence of ToBRFV on weed hosts. Identification of natural reservoirs of ToBRFV can help to develop management practices focused on weed plant species to prevent ToBRFV transmission. The extent to which ToBRFV survives in diverse alternate weed host species outside tomato growing seasons in different world regions requires further research in order to establish the risk associated with the possible contribution of weeds as a reservoir for primary infections in tomato crops.

Serum Endocan Levels and Subclinical Atherosclerosis in Behçet's Syndrome.

Background and AimBehçet disease (BD) is a rare chronic relapsing-remitting inflammatory systemic vasculitis. BD patients were reported to have marked acceleration of subclinical atherosclerosis (SCA). Endocan is a soluble proteoglycan mainly secreted by the activated endothelium. The present study aimed to assess the relation between serum endocan levels and SCA in BD patients.Subjects and MethodsThe study included 40 adult BD patients in addition to twenty age- and sex-matched healthy controls. BD was diagnosed according to International Study Group criteria. Upon recruitment, all participants were subjected to careful history taking and thorough clinical examination. BD activity was assessed using Behçet Syndrome Activity Score. Measurement of serum endocan was performed using quantitative double-antibody sandwich ELISA kit. CIMT measurement was done using B-mode ultrasound.ResultsComparison between patients and controls regarding serum endocan levels revealed significantly higher endocan levels in BD patients [median (IQR): 155.0 (69.3–610.0) versus 73.8 (51.9–94.6)]. Using ultrasound assessment, SCA was found in 14 BD patients (35.0%). Comparison between patients with SCA and patients without regarding the clinical and laboratory data revealed that the former group had significantly higher CRP [median (IQR): 36.5 (26.8–43.5) versus 21.0 (11.8–26.8) mg/dL, p < 0.001] and endocan [median (IQR): 622.0 (107.4–974.8) versus 104.5 (64.0–342.0) mg/dL, p = 0.004] levels. Logistic regression analysis recognized endocan [OR (95% CI): 1.0 (1.0–1.012), p0.035] levels as significant predictor of SCA in multivariate analysis.ConclusionThe present study identified the clinical value of serum endocan levels as a possible early marker of vascular involvement in BD patients.

The Effects of Phellodendron Decoction on Wound Healing of Anal Fistula after Anal Fistulotomy.

Objective To analyze the therapeutic effect of Compound Phellodendron decoction on wounds after anal fistulotomy. Methods 100 patients with anal fistula who underwent anal fistulotomy from April 2019 to April 2021 were included in the study group and control group according to the random number table method. 50 patients in the study group were treated with Compound Phellodendron decoction by fumigation and sitting bath, while warm water replaced Compound Phellodendron decoction in the control group. Perianal pain, wound edema, and exudation were scored on postoperative days 3 and 7, and wound healing time was recorded. Interleukin-2(IL-2), IL-5, IL-6, and IL-12 were measured via a double-antibody sandwich enzyme-linked immunosorbent assay. Hematoxylin-eosin (HE) staining and immunofluorescence were used to quantitatively analyze the capillary number and CD4+ and CD8+ lymphocytes in granulation tissue on postoperative days 7. Results The scores of pain, edema, and exudation in 2 groups on postoperative day 7 were lower than those on the 3rd postoperative day. Compared with the control group, the pain, edema, and exudation scores in the study group were decreased, and the wound healing time was shortened; the expressions of IL-2 and IL-12 in the study group were significantly increased, while the expressions of IL-5 and IL-6 were decreased; the number of capillaries and CD4+ lymphocytes in the study group was increased, while the number of CD8+ lymphocytes was decreased. Conclusion Compound Phellodendron decoction had efficacy in promoting wound healing, reducing complications, and changing lymphocyte aggregation and alleviating local inflammatory response.

An immunodetection assay developed using cobra cytotoxin-specific antibodies: Potential diagnostics for cobra envenoming

Cobra (Naja spp.) envenoming is a life-threatening medical emergency, and a correct diagnosis is crucial to initiating timely and appropriate antivenom treatment. However, snakebite diagnostics remain unavailable in Southeast Asia. This study, therefore, developed an immunodetection assay with a potential diagnostic application for cobra envenoming. The cytotoxin of Naja kaouthia (Thai Monocled Cobra) (Nk-CTX) was purified from its venom to produce CTX-specific antibodies in rabbits and chickens. A double-antibody sandwich enzyme-linked immunosorbent assay was developed using the purified anti-Nk-CTX antibodies (immunoglobulin G and immunoglobulin Y), and its selectivity, specificity, and sensitivity for the venoms of five major cobra species in Southeast Asia (N. kaouthia, Naja sumatrana, Naja sputatrix, Naja siamensis, and Naja philippinensis) were studied. The results showed the immunoassay discriminates cobra venoms from other species commonly implicated in snakebites in Southeast Asia, i.e., the Malayan Krait, Many-banded Krait, King Cobra, Eastern Russell's Viper, Malayan Pit Viper and White-lipped Pit Viper. The immunoassay has a high sensitivity for the five cobra venoms, with detection limits (LoD) ranging from 0.6 to 2.6 ng/ml. Together, the findings suggest the potential diagnostic application of the cytotoxin immunoassay for cobra envenoming. The immunoassay was found to exhibit high immunoreactivity toward ten Asiatic cobra venoms (absorbance > 1.5), in contrast to African cobra venoms with low immunoreactivity (absorbance < 0.9). Considering the varying CTX antigenicity between Asiatic and African cobras, the immunoassay for African cobras should utilize antibodies produced specifically from the cytotoxins of African cobra venoms.

Antibodies for the Coat Protein of Cotton Leafroll Dwarf Virus Detect Commelina sp. as an Intermediary Host for Cotton Blue Disease.

The cotton blue disease, caused by the cotton leafroll dwarf virus (CLRDV), leads to dwarfism, leaf rolling, and production loss in susceptible cotton varieties. To develop an enzyme-linked immunosorbent assay (ELISA) test to detect the virus in cotton and weeds, peptides based on the coat protein were used to produce polyclonal (α-GQE, α-PRN, and α-INK) and monoclonal (α-GQE, α-PRN, and α-NKF) antibodies. All six were tested as capture antibodies, and polyclonal α-GQE and the monocle onal α-NKF were labeled with the enzyme alkaline phosphatase and used as detection antibodies for a double antibody sandwich (DAS) ELISA method, in which p-nitrophenyl phosphate was added and measured by absorbance at 405 nm. The DAS-ELISA sandwich was efficient in discriminating between healthy and diseased plant extracts. The ELISA methodology detected the virus in the weeds Commelina sp., which was confirmed by RT-PCR. The monoclonal antibodies may be used to develop other diagnostic procedures.

Protective effect of ivabradine on mice with viral myocarditis and its mechanism

Viral myocarditis (VMC) is a type of cardiovascular disease caused by viral infection of myocardial cells characterized by myocardial interstitital inflammatory cell infiltration, myocardial fiber necrosis or figrinolysis. Ivabradine (IVA) is a commonly known drug used to control heart rate, resist inflammation and oxidative stress, particularly in VMC. Here, we have tried to evaluate the protective effects of IVA on mice with VMC and understand the possible mechanism behind this process. In order to complete the study, eighty male mice aged 6 weeks old were randomly divided into normal control, VMC model, low-dose IVA (L-IVA) and high-dose IVA (H-IVA) groups. Half an hour after modeling, IVA aqueous solution was administered intragastrically into L-IVA and H-IVA groups at 5 mg·kg-1·d-1 and 20 mg·kg-1·d-1, respectively for 14 consecutive days. Another 120 mice of the same batch were grouped and treated as described above. At 7 and 14 d, 6 mice in each group were sacrificed to obtain blood and heart samples. Body weight/heart weight (BW/HW) was calculated, hematoxylin-eosin staining was performed to observe the pathological changes of myocardium, and the level of cardiac troponin I (cTnI) was measured by ELISA. Related kits were employed to measure superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px) and catalase (CAT) activites in myocardial homogenate, and the levels of interleukin-6 (IL-6), IL-18, IL-1β and tumor necrosis factor-α (TNF-α) were determined by double-antibody sandwich ELISA. TUNEL assay was performed to detect the apoptosis of myocardial cells. The protein expressions of Bcl-2, Bax and Caspase-3 in myocardial cells were measured by Western blotting. Compared to the VMC model group, the heart/body weight ratio, myocardial pathological score, cTnI level, MDA activity, and levels of IL-6, IL-18, IL-1β and TNF-α were found decreased, while survival rate and activity of SOD, GSH-Px and CAT increased in L-IVA and H-IVA groups (P &lt;0.05). The apoptosis rate of myocardial cells declined in L-IVA and H-IVA groups (P &lt;0.05), especially in H-IVA group. IVA downregulated the protein expressions of Bax and Caspase-3 and upregulated that of Bcl-2 (P &lt;0.05). Thus, it has been found that IVA elevates the survival rate of VMC mice and relieves myocardial damage possibly by enhancing antioxidant capacity and modulating apoptosis-related proteins to suppress apoptosis.

Investigation of the location and secretion features of Candida albicans enolase with monoclonal antibodies

PurposeThe glycolytic enzyme enolase plays important role in the pathogenesis of Candida albicans infection and has been also considered as a promising molecular marker for the diagnosis of invasive candidiasis. This study aimed to investigate the location and secretion features of Candida albicans enolase (CaEno) with a couple of specific monoclonal antibodies (mAbs).MethodsTwo mAbs named 9H8 and 10H8 against CaEno were generated by fusing SP2/0 myeloma cell with the spleen lymphocytes from CaEno immunized mice. The specificity of the mAbs was then validated by Western blot and liquid chromatography-mass spectrometry (LC–MS/MS). A diverse set of experiments were conducted based on the pair of mAbs which involved immunohistochemical staining analysis, whole cell enzyme-linked immunosorbent assay (ELISA), double antibody sandwich ELISA, and confocal microscopy to analyze the possible location and secretion features of CaEno.ResultsCaEno is abundantly expressed in the cytoplasm of C. albicans blastospores and is distributed in a ring-shaped pattern along the cell wall. CaEno appeared in the hyphal C. albicans as just a “mushroom” form. CaEno was found to be weakly expressed on the surface of blastospores but constantly expressed at various stages of growth. CaEno concentrations in C. albicans blastospores culture supernatant are considerably higher than in C. albicans hyphae culture supernatant. The dynamic changes of supernatant CaEno concentration in blastospores and hyphal C. albicans exhibit distinct features, although both appear to be associated with the C. albicans growth state. When cultivated under normal circumstances, however, no apparent CaEno degradation was seen in the cell-free supernatant.ConclusionOur results implied that CaEno was constantly expressed on the cell surface and its secretion features varied according to the growth stage of C. albicans. However, further experimental and theoretical studies are needed in future to identify the specific mechanisms by which this phenomenon can arise.

Prognostic value of serum CEA and CA19-9 levels in pancreatic ductal adenocarcinoma.

The present study investigated the associations of serum carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) levels with clinicopathological variables and survival outcomes in Libyan patients with pancreatic ductal adenocarcinoma (PDAC). The clinicopathological variables of 123 patients with PDAC registered at the National Cancer Institute in Misurata, Libya, between 2010 and 2018 were retrospectively analyzed. Blood samples from these patients were analyzed for serum CEA and CA19-9 levels before treatment by electrochemiluminescence immunoassay (double antibody sandwich ELISA) on a Roche cobas e 602 modules. The relationships between CA19-9 and CEA serum levels with clinicopathologic variables and survival outcomes were analyzed using the Kaplan-Meier method, log-rank test and Cox regression analyzes. Cut-off values for serum CEA and CA19-9 levels were 5 ng/ml and 400 U/ml, respectively. The median serum levels of all patients with PDAC for CEA and CA19-9 were 8 ng/ml (1.1-377 ng/ml) and 389 U/ml (1-10,050 U/ml), respectively. Tumors with higher serum CEA and CA19-9 levels were found in 63 and 48% of patients, respectively. Higher CEA and CA19-9 serum levels were significantly associated with more indicators of a malignant phenotype, including a surgically unresectable tumor, unevaluable lymph nodes, advanced stages and distant metastases. Regarding survival, patients with higher serum levels of the biomarkers CEA and CA19-9 had shorter overall survival rates (P<0.016 and (P<0.014, log-rank, respectively) and lower disease-free survival rates (P<0.002 and P<0.0001, log-rank, respectively). The present study demonstrated significant clinical and prognostic value of serum levels of biomarkers CEA and CA19-9 for Libyan patients with PDAC. Moreover, patients with PDAC with higher serum CEA and CA19-9 levels had more aggressive tumors, higher rates of disease recurrence and shorter overall survival rates and thus required more vigilant follow-up. Further multinational studies with larger PDAC cohorts are warranted to confirm these findings in terms of improved clinical decision making, more effective management and improved survival.

Broad-spectrum resistance against multiple PVY-strains by CRSIPR/Cas13 system in Solanum tuberosum crop

ABSTRACT Potato virus Y (PVY) is a deadly environmental constraint that damages productivity of potato (Solanum tuberosum) around the globe. One of the major challenges is to develop resistance against PVY. Emerging clustered regularly short palindromic repeat (CRISPR)/Cas systems have the potential to develop resistance against PVY. In the current research, CRISPR-Cas13 has been exploited to target multiple strains of PVYN, PVYO, and PVYNTN. Multiple genes PI, HC-Pro, P3, Cl1, Cl2, and VPg genes of PVY were targeted by CRISPR/Cas13a. Multiplex gRNA cassettes were developed on the conserved regions of the PVY-genes. Three independent CRISPR/Cas13 transgenic potato lines were developed by applying an optimized concentration of trans-ribo zeatin and indole acetic acid at callus development, rooting, and shooting growth stages. The level of resistance in transgenic plants was confirmed through double-antibody sandwich enzyme-linked immunosorbent assay and real-time quantitative PCR. Our results have shown that efficiency of PVY inhibition was positively correlated with the Cas13a/sgRNA expression. Finding provides the specific functionality of Cas13 with specific gRNA cassette and engineering the potential resistance in potato crop against multiple strains of PVY.

Detection of Cucumber mosaic virus on Solanum lycopersicum L. and Capsicum annuum L. in the Western region of Cameroon

The Cucumber mosaic virus (CMV) is of major concern to large and small-scale growers of tomato and pepper in the Western region of Cameroon. It causes loss in quantity and quality of produce. Here, we described the detection of CMV and symptomatology in infected tomato and pepper grown in the Western region of Cameroon using a serological method. To detect CMV, 180 leaves of each species were tested. Leaves were collected from the surveyed fields were screened initially for the CMV using Double Antibody Sandwich Enzyme Linked Immunosorbent Assay (DAS-ELISA) using polyclonal antibodies of CMV. To draw up the list of the symptoms, 300 symptomatic leaves were collected, 150 leaves per species and DAS-ELISA was used to confirm the presence of CMV in the leaf sample. The proportion of infected leaves among the total sample determined. The prevalence of CMV on tomato and pepper were 37.22% and 78.33% respectively. In conclusion, CMV infects tomato and pepper in the West Region of Cameroon to a varying extent. Also, assessing only symptom on the two crops is not enough for the determination of the health status of both crops with respect to CMV. As a result of this study, there is a need to sensitize tomato and pepper farmers about CMV and to provide them with basic vector-management strategies.

Protective effect of symmetrical N‐heterocyclic 1,3,4‐oxadiazole and 1,3,4‐thiadiazole derivatives against Pepino mosaic virus of tomato

AbstractPepino mosaic virus (PepMV) is a highly infectious potexvirus, which has presently become a major pathogen for tomato crops worldwide. Since there is no effective method control for PepMV, strict preventive hygienic measures and cross‐protection have been established. However, the use of mild isolates to protect against aggressive isolates provides opportunities for interaction with other microorganisms, which under certain conditions may worsen disease symptoms. Thus, alternative control methods are needed. In this study we aimed at the development of chemical control against PepMV based on the use of heterocyclic compounds including symmetrical 2,5‐disubstituted 1,3,4‐oxadiazoles, symmetrical 2,5‐disubstituted 1,3,4‐thiadiazole and dihydrotetrazine derivatives. Results showed that spray application of three 1,3,4‐oxadiazole derivatives (OH‐Oxa, CH3‐Oxa, NO2‐Oxa) and of the derivative of 1,3,4‐thiadiazole (OH‐Thia) resulted in reducing the main symptoms of PepMV in tomato leaves, conversely to the dihydrotetrazine carboxylic acid. Double‐antibody sandwich enzyme‐linked immunosorbent assay (DAS‐ELISA) revealed that PepMV was not detected from tomato plants pre‐treated with the derivatives conferring elevated protection. Moreover, the protective ability of OH‐Oxa, CH3‐Oxa, NO2‐Oxa and OH‐Thia was closely related to the enhancement of the activity of antioxidant enzymes upon infection with PepMV.

Synthesis of Coumarin Derivatives: A New Class of Coumarin-Based G Protein-Coupled Receptor Activators and Inhibitors.

To expand the range of daphnetin-based inhibitors/activators used for targeting G protein-coupled receptors (GPCRs) in disease treatment, twenty-five coumarin derivatives 1–25, including 7,8-dihydroxycoumarin and 7-hydroxycoumarin derivatives with various substitution patterns/groups at C3-/4- positions, were synthesized via mild Pechmann condensation and hydroxyl modification. The structures were characterized by 1H NMR, 13C NMR and ESI-MS. Their inhibition or activation activities relative to GPCRs were evaluated by double-antibody sandwich ELISA (DAS–ELISA) in vitro. The results showed that most of the coumarin derivatives possessed a moderate GPCR activation or inhibitory potency. Among them, derivatives 14, 17, 18, and 21 showed a remarkable GPCR activation potency, with EC50 values of 0.03, 0.03, 0.03, and 0.02 nM, respectively. Meanwhile, derivatives 4, 7, and 23 had significant GPCR inhibitory potencies against GPCRs with IC50 values of 0.15, 0.02, and 0.76 nM, respectively. Notably, the acylation of hydroxyl groups at the C-7 and C-8 positions of 7,8-dihydroxycoumarin skeleton or the etherification of the hydroxyl group at the C-7 position of the 7-hydroxycoumarin skeleton could successfully change GPCRs activators into inhibitors. This work demonstrated a simple and efficient approach to developing coumarin derivatives as remarkable GPCRs activators and inhibitors via molecular diversity-based synthesis.

Antioxidant Effect of Thioredoxin and Vitamin D3 in Peritoneal Dialysis Patients.

Background Among the chronic diseases, chronic kidney failure is one of diseases that have the most difficulty in coping with oxidative stress due to the deterioration of the antioxidant system balance in the body. Beyond being a vitamin, 1α,25-dihydroxycholecalciferol (vitamin D3) is a molecule that positively or negatively affects many enzymes which are in protein structures. Thioredoxin (TRX), which has an important role in the antioxidant system, is one of these proteins. By conducting this study, we wanted to emphasize the role of vitamin D3 in reducing the oxidative stress load on patients undergoing peritoneal dialysis (PD) via serum TRX level measurement. Methods In this study, we evaluated the medical treatments of 69 PD patients who were followed up routinely. The patients were divided into 2 groups according to whether they used vitamin D3 or not. 49 of our patients were using vitamin D3. While requesting routine laboratory tests, we reserved a separate serum sample to measure serum TRX levels by double-antibody sandwich enzyme-linked immunosorbent assay for all patients. Results Only one parameter has a significant statistical relationship with serum TRX level and the treatment protocol. The serum TRX level was significantly higher (211,62 U/l ± 314,46) in the group receiving vitamin D3 compared to the group which is not using Vitamin D3 (101,63 U/l ± 215,03) (p < 0,006). Conclusion This study highlights the importance of appropriate dose of vitamin D3 replacement especially in PD patients who are under intense oxidative stress compared to healthy individuals.

Establishment of DAS-ELISA for the detection of antigenic changes in glycinin after heat processing

In this study, a double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) method was established to detect the antigenic changes of thermally processed products containing glycinin. The proposed DAS-ELISA method used heat-treated antigen-absorbing antiserum as the coating antibody, and horseradish peroxidase (HRP)-labeled rabbit anti-glycinin polyclonal antibody as the detection antibody. The specificity test results which were obtained using the proposed method indicated that good specificity had been achieved. The cut-off value was 0.388, and the LOD was determined to be 19.53 ng/mL. The coefficient of variation was less than 5.25% (intra-day) and 9.50% (inter-day). In this study's milk powder addition test, the recovery rate of the glycinin ranged between 83.65% and 90.13%. The established DAS-ELISA method was also used to detect soybean thermal processing products, such as soy sauce, steamed fish and soy sauce, soybean paste, beef sauce, soy milk powder, and tofu. The results showed that the OD450 values of the aforementioned products were lower than the OD450 values of the glycinin in defatted soybean flour. Therefore, it was indicated that the above products has undergone different degrees of thermal processing. In other words, the majority of the epitopes of glycinin in the products had been destroyed by the thermal processing and could not be combined with heat-treated antigen-absorbing antiserum.

Screening of sweet potato feathery mottle virus resistant sweet potato (Ipomoea batatas L., Lam.) cultivars in Kebbi State, Nigeria

Sweet potato is a food security crop because of its ability to withstand adverse climatic conditions. This security, however, is being threaten by viral diseases and use of resistant cultivars remain the best management. This research was conducted to screen cultivars of sweet potato against sweet potato feathery mottle virus (SPFMV) infection. The treatments consisted of five cultivars coded as CV1, CV2, CV3, CV4 and CV5, respectively. The asymptomatic experimental plants were established, maintained under screen house conditions and graft-inoculated using infected vines which were tested SPFMV positive using both Double Antibody Sandwich Enzyme-Linked Immunosorbent Assay (DAS-ELISA) and Polymerase Chain Reaction (PCR). The experiment was laid out in Completely Randomized Design (CRD) and replicated three times. Results obtained from disease incidence and symptom severity indicated that there was significant difference (P &lt;0.05) among cultivars in their reaction to SPFMV infection. CV2 had the highest mean disease incidence (60.67%) while, CV3 had the lowest mean disease incidence (36.67%). CV1, CV3, and CV5 have the same lowest mean symptom severity score of 2.00 while, CV2 had the highest mean severity score of 4.00. Based on the reaction of the cultivars after inoculation, it could be concluded that, all the cultivars screened were susceptible to SPFMV but CV1, CV3 and CV4 cultivars have some degree of resistance to SPFMV infection and were therefore recommended for use by the farmers in the study area. This is the first research that screened sweet potato cultivars for resistance to SPFMV in Kebbi State, Nigeria.

Evaluation of serum hepcidin level in acne vulgaris patients

Background: Acne vulgaris is a pilosebaceous unit inflammatory condition. Although acne is not life-threatening, it can cause psychological issues such as low self-esteem and anxiety in people who suffer from it. Acne scarring is a common and unpleasant consequence of acne vulgaris. Predicting scarring liability can modify treatment protocol and aid in the prevention of such disfigurement. Methods: This study included 82 participants divided into 3 groups: 26 patients with acne vulgaris without scars, 36 patients with acne vulgaris with scars and 20 control. Serum hepcidin level was assessed by a commercially available double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) kit. Results: Serum hepcidin is significantly higher in patients suffering from acne without acne scars than patients suffering from acne with acne scars (p &lt; 0.05). serum hepcidin is significantly higher in the control group than patients with active acne with scars (p &lt; 0.05). There was a statistically significant difference between serum hepcidin level in control versus severe acne scars (p &lt; 0.05). Serum hepcidin level acts as a predictor of occurrence of acne scars and occurrence of severe acne scars. Serum hepcidin level can be used as an indication for beginning aggressive and specific acne treatment as systemic retinoid.

Risk Prediction of Coronary Artery Stenosis in Patients with Coronary Heart Disease Based on Logistic Regression and Artificial Neural Network

Objective Coronary heart disease (CHD) is considered an inflammatory relative disease. This study is aimed at analyzing the health information of serum interferon in CHD based on logistic regression and artificial neural network (ANN) model. Method A total of 155 CHD patients diagnosed by coronary angiography in our department from January 2017 to March 2020 were included. All patients were randomly divided into a training set (n = 108) and a test set (n = 47). Logistic regression and ANN models were constructed using the training set data. The predictive factors of coronary artery stenosis were screened, and the predictive effect of the model was evaluated by using the test set data. All the health information of participants was collected. Expressions of serum IFN-γ, MIG, and IP-10 were detected by double antibody sandwich ELISA. Spearman linear correlation analysis determined the relationship between the interferon and degree of stenosis. The logistic regression model was used to evaluate independent risk factors of CHD. Result The Spearman correlation analysis showed that the degree of stenosis was positively correlated with serum IFN-γ, MIG, and IP-10 levels. The logistic regression analysis and ANN model showed that the MIG and IP-10 were independent predictors of Gensini score: MIG (95% CI: 0.876~0.934, P < 0.001) and IP-10 (95% CI: 1.009~1.039, P < 0.001). There was no statistically significant difference between the logistic regression and the ANN model (P > 0.05). Conclusion The logistic regression model and ANN model have similar predictive performance for coronary artery stenosis risk factors in patients with CHD. In patients with CHD, the expression levels of IFN-γ, IP-10, and MIG are positively correlated with the degree of stenosis. The IP-10 and MIG are independent risk factors for coronary artery stenosis.

Assessment of Two Different Glucagon Assays in Healthy Individuals and Type 1 and Type 2 Diabetes Patients.

Methods for glucagon analysis suffered in the past from lack of specificity and a narrow sensitivity range, which has led to inaccurate results and to the suggestion that type 1 diabetes (T1D) and type 2 diabetes (T2D) patients have elevated fasting glucagon levels. However, the availability of more specific and more sensitive methods to detect intact glucagon has shown that actual glucagon levels are lower than previously assumed. This study aimed to characterize fasting plasma glucagon levels in healthy individuals and T1D and T2D patients with two different glucagon assays. The study included 20 healthy individuals, 20 T1D and 20 T2D patients. Blood was collected under fasting conditions. A double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) and a conventional radioimmunoassay (RIA) were used. A significant difference in fasting glucagon levels between healthy individuals and T2D was observed by ELISA, but not by RIA. ELISA also yielded lower glucagon levels in healthy individuals than in T1D and T2D patients which RIA did not. RIA produced significantly (p = 0.0001) higher overall median glucagon values than ELISA in a pooled analysis. These results underline the notion that the choice of selective laboratory methods is highly relevant for mechanistic endocrine research.

Detection of orthotospoviruses in medicinal plants in China

AbstractThe genus Orthotospovirus contains a variety of viruses that infect more than 1000 plant species of 82 families. Nine medicinal plant species were found to exhibit symptoms that are typical of plant viral infections. Foliar samples of diseased plants were taken, and double‐antibody sandwich ELISA (DAS‐ELISA) and reverse transcription polymerase chain reaction (RT‐PCR) analyses were used to detect the viruses in the diseased plants. The results revealed that Capsicum chlorosis orthotospovirus (CaCV) was present in Salvia miltiorrhiza and Valeriana jatamansi. Impatiens necrotic spot orthotospovirus (INSV) was identified in Plantago asiatica, Scrophularia ningpoensis and Iris tectorum. Tomato spotted wilt orthotospovirus (TSWV) was detected in Perilla frutescens, while Tomato zonate spot orthotospovirus (TZSV) was found in Codonopsis spp., Marsdenia tenacissima and Dipsacus asper. Phylogenetic tree analysis also reflected the high consistency of sequences. To our knowledge, this is the first report of CaCV in Salvia miltiorrhiza and Valeriana jatamansi and INSV in Plantago asiatica, Scrophularia ningpoensis and Iris tectorum. This is also the first report of TSWV infecting Perilla frutescens and TZSV infecting Codonopsis spp., Marsdenia tenacissima and Dipsacus asper.