The purpose of the present study was to define the period of time in which aerobic training does not increase further serum S-Klotho levels in untrained young adult males, and to examine the relation between plasma S-Klotho concentration and maximal oxygen uptake (VO2max).
Methods: Sixty (60) untrained subjects (27.05±1.1 years) were divided into 2 groups, both exercised six months 4×wk-1 for the duration of 45 min×session. One group (LTI) exercised below the anaerobic threshold at 40-50% of VO2max, while the second group (HTI) worked above the anaerobic threshold at 65-70% of VO2max. Testing sessions were performed at 0, 2, 4, and 6 months. Blood samples were drawn after overnight fasting; S-Klotho was analyzed using an ELISA kit.
Results: Following 2 and 4 months, significant (p≤0.05) increases were noted in the HTI group, at the fourth testing session, S-Klotho leveled off. In the LTI group, S-Klotho remained almost unchanged. Findings of the present study, support emerging evidence suggesting that a relation between plasma S-Klotho concentration and VO2max exists.
Conclusion: Data suggest that increases in S-Klotho is tidally associated with VO2max levels. In addition, the S-Klotho increase levels-off following 4 months of aerobic training. Exercising below the anaerobic threshold does not increase VO2max and thus, does not increase S-Klotho.
Botulism is the disease caused by botulinum neurotoxins. It is produced by an obligate anaerobic bacteria called Clostridium botulinum. There is no immuno-detection system available in the world for the detection of C. botulinum. Secretory proteins of cooked meat media grown C. botulinum type B were extracted by TCA precipitation method. Polyclonal antibodies were generated against secretory proteins. Cytokine profiling of secretory proteins were done. An immunodetection system was developed to detect the C. botulinum type B using Secretory proteins of C. botulinum type B.
Objective: to determine the incidence of HBV and HCV in pediatric ward.
Sitting: 2ed March teaching hospital, sebha Libya.
Materials and Methods: this was a prospective hospital base study of pediatric cases admitted to 2ed March teaching hospital during a period from March 2018 to February 2019. Pediatric cases were studied for the incidence of HBsAg and HCV Ab by ELISA, Rapid technique. The positive result was confirmed with line immuno-assay.
Results: the study showed positive HBsAg in 12 patients and HCV in 2 cases out 25 cases represented with acute hepatitis from a total of 1763 pediatric cases were submitted in this study, with incidence rate of 0.68% and 0.11% respectively.
Conclusion: the incidence of HBV and HCV are low in Sebha, therefore active program need to be applied to control the spread of infection among the population.
Infectious bursal disease (IBD) considered as one of the major viral diseases threatening the poultry industry worldwide. The causative agent of the IBD is Infectious bursal disease virus (IBDV) which replicates in developing B lymphocytes in the bursa of Fabricius leading to its destruction and bursal inflammation. In this study, we investigated a technology to produce therapeutic recombinant antibodies against IBDV in bacteria by constructing a bacterial displayed recombinant scFv library from immunized chickens, followed by screening the scFv library by fluorescence activated cell sorting (FACS) with FITC-labeled VP2. Twelve VP2-binding scFv clones with unique sequences were obtained, with overall amino acid homology of 81.53%. The complementarity determining region (CDR) 3 in the heavy chain displayed the lowest homology, while the amino acid sequences in framework regions and CDR2 of both chains and CDR1 of the heavy chain are relatively conserved. Twelve VP2-binding scFv clones were expressed in E.coli and purified through denaturation and denaturation of inclusion bodies. Our ELISA results showed that all scFvs exhibited binding ability and specificity to VP2 and various IBDV strains. In addition, two scFvs showed significant neutralizing activity to IBDV (B-87 strain) as these scFvs inhibited cytopathic effect of chicken embryo fibroblast (DF1) caused by IBDV. In conclusion, our study provides a lead candidate for further development of therapeutic antibodies for IBDV infection.
The insufficiency of interferon production and the cytokine imbalance in patients with atopic dermatitis, especially in combination with persistent herpes virus infection, has been identified. The expediency of the use of interferon inducer Cycloferon in the treatment of chronic atopic dermatitis has been shown.
By researching the factors related to exposure to indoor and outdoor allergens, such seasons, climate changes and particulate matter, allergists can screen the sensitization profile of individuals according to their exposures and conduct preventive treatment and individualized immunotherapy. Molecular allergology has improved aerobiological screening of allergenic components toward more specific results on allergic exposure, sensitization, and symptoms [1,2]. The Enzyme-Linked Immunosorbent Assay (ELISA) is a colorimetric enzyme immunoassay technique used to quantify soluble substances such as proteins, peptides, antibodies, and hormones. Due to its high sensitivity and specificity, ELISA can quantify substances at low concentrations, such as allergens .
Benjamín Guix*, Teresa Guix, Marco Panichi, Ines Guix, Iván García, Carles Llebaría, Nicolás Achkar, Luis Quinzaños, Hamza Sentisi, Jose Luís Enríquez, Ana Galván, Cristina Pérez-Sánchez, Víctor González and Carmen León
Introduction: Serology (antibody) tests for the SARS-CoV-2 have been proposed as an instrument to inform health authorities about immunization during the COVID-19 pandemic. As there is a significant part of the population that may have some degree of immunity, it is of great interest to communicate the immunization results obtained in the first 500 healthcare workers (HCW), patients and relatives tested in a community-based Oncological Center.
Materials and methods: Between April 9th, 2020 and May 8th, 2020, a group of healthcare workers (HCW), their families, and general public who had had the COVID-19 or had been in close contact with confirmed cases of COVID-19 were screened for IgG SARS-CoV-2 antibodies. The tests were carried out in a rigorous manner, strictly following the guidelines approved by the Spanish Ministry of Health (Ministerio de Sanidad).
Results: The major objective of this study was to determine the proportion of asymptomatic infected individuals and those who had already secreted IgG against SARS-CoV-2 in our cancer treatment center or in the community of Barcelona. Patients were tested with PCR, Rapid diagnostic test (RDT) or enzyme-linked immunoabsorbent assay (ELISA). A total of 521 participants were tested, 206 with RDT and 315 with ELISA, 59 (11,32%) resulted positive to SARS-CoV-2.
Conclusion: RDT and ELISA proved to be effective and sensible enough to determine the extent of SARS-CoV-2 immunization in a community-based oncological center. The degree of immunization reached is nowadays far away from what can be considered desirable for a herd immunization.
Purpose: To evaluate the levels of salusin-beta (β-SAL) in the serum in patients with age-related macular degeneration (ARMD).
Methods: Our study was designed as a controlled comparative clinical study. The β-SAL levels in serums of age and sex-matched 20 healthy volunteers as controls (Group 1), 20 patients with dry-age related macular degeneration (d-ARMD) (Group 2) and 20 patients with wet-age related macular degeneration (w-ARMD) (Group 3) were measured with the enzyme-linked immunosorbent assay (ELISA) method.
Results: In our study, it was found that age and gender didn’t show a statistically significant difference among the study groups (p > 0. 05). The mean serum β-SAL levels in Group 1, Group 2 and Group 3 were 1372,17 ± 1126.69 pg/mL; 1423,71 ± 1196.84 pg/mL and 940,57 ± 1092.05 pg/mL, respectively. Although the meanβ-SAL levels in w-ARMD seem numerically lower than both the control and d-ARMD groups, this difference among the study groups was not statistically significant (p > 0.05).
Conclusion: Our study suggests that β-SAL levels in the patients with ARMD and healthy controls were not different than each other. Further studies with large numbers may reveal possible relationships between β-SAL and ARMD.
Background and objectives: New AKI biomarkers (on the top of it NGAL biomarker) have demonstrated better performance for prediction of AKI in critically ill patients with heterogeneous illness. Renal angina index was recently reported to enhance prediction of severe AKI at the time of intensive care unit admission. This study tested the hypothesis that incorporation of uNGAL in patients with renal angina improves the prediction of severe AKI.
Design, setting, participants & measurements: In our study 53critically ill children admitted to the pediatric intensive care unit in Zagazig university hospital, Measurement of urine neutrophil gelatinase– associated lipocalin (uNGAL) was determined individually by ELISA kit and in combination with the RAI which is calculated in each critically ill child for severe AKI. Statistical analysis was done for these data.
Results: Individual uNGAL demonstrated marginal discrimination for severe AKI (area under curve [AUC]: NGAL, 0.877), little higher than prediction by RAI (AUC=0.847). Incorporation of uNGAL significantly added to the renal angina index AKI prediction (AUC=0.847, increased to 0.893).
Conclusion: This study shows that incorporation of uNGAL into the RAI improves detection ability of severe AKI in critically ill children.
Pigeonpea is one of the important legume crops with high protein content and nutritional traits. It has enormous potency for its widespread adoption by farming communities. It is affected by various kinds of biotic and abiotic stresses. In the context, of biotic stresses Sterility mosaic disease (SMD) is one of the severe diseases in pigeonpea which ultimately lead to the drastic yield loss. The virus belongs to the genus Emaravirus, family- Fimoviridae. SMD is associated with two diverse types of Emaravirus, Pigeonpea sterility mosaic virus1 (PPSMV-1) and Pigeonpea sterility mosaic virus 2 (PPSMV-2). It is transmitted by the mite (Aceria cajani), mainly environmental contributing to the feasibility for the mites for the inoculation of the virus. The SMD is mainly governed by two genes SV1 that includes the dominant allele and serves as an inhibitory action on the resistance of the SV2. Methods for identification of the virus include RT-PCR, DIBA and ELISA using alkaline phosphatase or penicillinase. To control SMV disease farmers generally adopted intercropping methods. There are few potential drugs have been identified for the administration of the disease such as 0.1% Fenazaquin, Dicofol, Imidacloripid, Carbosulfan; Spiromesifin includes the inhibition of the mite inoculation on the pigeonpea plant. The present review describes compressive and systematic insights on SMV protein targets and potential drugs that could be utilized as the presumed drug targets for the finding of true drugs against the SMD in pigeonpea.
Mast cells play a central role in the genesis and modulation of allergic and inflammatory responses. The general aim of the present work was to study the interaction between mast cells and the most common additives approved for use in foods. Dose-response studies about the effect of the main food additives (tartrazine, sodium bisulphite and sodium benzoate) on mast cell degranulation were carried out. Rat peritoneal mast cells were incubated with: 1) buffer solution or 2) stimulus. The stimuli were tartrazine, sodium benzoate, sodium bisulphite and the calcium ionophore A23187. A23187 was used as a reference mast cell secretagogue. Different doses and combinations of food additives were used. The viability of the mast cells was evaluated with trypan blue. In the incubation solutions, the release of β-hexosaminidase was quantified by colorimetric reaction and ELISA plate reader. The remaining β-hexosaminidase concentration (not released) was studied in the cells after the incubations, and morphology of the mast cells was analyzed by light microscopy with toluidine blue stain. The food additives tartrazine, sodium benzoate and sodium bisulphite did not stimulate the release of β-hexosaminidase from mast cells at any of the concentrations used. In contrast, tartrazine at concentrations of 0.1 μM and 1 μM, and sodium benzoate and sodium bisulphite at concentrations of 0.1 μM, 1 μM, 10 μM and 100 μM, significantly inhibited the basal release of β-hexosaminidase from mast cells. Considering these findings, we decided to determine the effect of these additives on the degranulation of mast cells induced by the calcium ionophore A23187. Sodium bisulphite inhibited mast cell activation induced by the calcium ionophore A23187 in this experimental model. The present study demonstrates that food additives of usual permitted use do not stimulate basal degranulation of mast cells in an in vitro model of peritoneal mast cells and that the additive sodium bisulphite inhibit mast cell activation induced by intracellular calcium increase. This food additive could represent an interesting alternative in the prevention of pathologies mediated by mast cells, as well as in the field of nutritional biochemistry.
Introduction and aim: Idiopathic nephrotic syndrome (INS) is the most common type of this disease during childhood. Minimal change nephrotic syndrome (MCNS) is the most common histopathological lesion (80 – 90%) of INS in children and about 90% of patients are steroid responsive, while congenital nephrotic syndrome is disorder that may be caused by several diseases. Intrauterine infections, especially CMV infection, have frequently been incriminated as etiological factors of secondary CNS. The aim of this research was to evaluate the frequency of CMV infection children with active nephrotic syndrome in our pediatric nephrology unit
Patients and methods: This descriptive (cross sectional) study was conducted in pediatric nephrology unit, Zagazig University Hospitals and included 60 patients WITH NS in activity; Participants were subjected to, Full history taking, Clinical examination; general & local, Routine laboratory investigations and Serum samples were tested for HCMV specific immunoglobulin G (IgG) and immunoglobulin M (IgM) using ELISA Kit.
Results: We found 100% of cases were IgG positive and 7/60 cases were IgM positive, There were no statistically significant differences between IgM positive-patients vs IgM-negative patients according to age, sex and first attack or relapsed NS, There were statistically significant differences between IgM positive-patients vs IgM-negative patients in blood laboratory data in decreases in HB (P=0.024) and serum urea nitrogen (P=0.04)
Conclusion: We concluded that serofrequency of cytomegalovirus infection in pediatric nephrology unit, Zagazig university hospitals during follow-up was 12% for cmv IgM and 100% for cmv IgG at ns children patients
Background: Hepatitis B virus infection is a major cause of liver associated morbidity and mortality with diverse spectrum of disease. It is estimated about 15% to 40% of patients with hepatitis B virus infection progress to chronic hepatitis and about 15% to 25% die from disease complications. The main aim of this study was to evaluate the serological and virological markers of patients with chronic hepatitis B virus infection to determine the natural history of chronic hepatitis B infection in the Eritrean setting.
Methods: A laboratory-based cross-sectional study was conducted on 305 patients with HBsAg positive who presented to Orotta National Referral Hospital, Halibet Hospital, Sembel Hospital and National Health Laboratory in Asmara, Eritrea from January 2017 to February 2019. Enzyme-linked immunosorbent assay was performed to detect hepatitis B serological markers (anti-HBc, HBsAg, anti-HBsAb, HBeAg and anti-HBeAg). Hepatitis B DNA viral loads and liver transaminase levels were determined. Data analysis was conducted using SPSS version 25.0.
Results: A total of 305 patients presented with HBsAg positive serology with a mean age of 41.3 (± 13.7) years ranging from 16 to 78 years. Males were 218 (71.5%) and females 87 (28. 5%).Anti-HBc was positive in 300 (98.4%), of which 293 (97.5%) were positive for HBsAg and 7 (2.3%) positive for anti-HBs. Among these 293 patients, 20 (6.8%) were HBeAg positive/anti-HBe positive, 242 (82.6%) HBeAg-negative/anti-HBe-positive and 31 (10.6%) were HBeAg negative/anti-HBe-positive. Detectable HBV DNA was found in 122(41.6%) of the 293 cases. Alanine transaminase was normal in 90% of HBeAg-positive and in 91.2% of HBeAg-negative patients. Hepatitis B DNA viral load was >2,000 IU/mL in 67 (22.86%) and >200,000 IU/mL level was more frequently detected in HBeAg positive (20.0%) compared to HBeAg negative (1.8%) subjects (p < 0.001).
Conclusion: This study shows predominance of HBeAg-negative and low replication phase of HBV infection among patients in Eritrea. It also documented that most patients had chronic infection with normal liver transaminase levels in the absence of biochemical signs of hepatitis. This study will provide a basis for therapeutic evaluation of patients and planning national treatment guidelines in the Eritrean setting.
Phytosanitary inspectors play an important role in diagnosing diseases in foreign plant material. However, some deficiencies have been detected in the detectionc ausing the entrance of many microorganisms. Therefore, it was of great interest to detect the presence of Clavibacter michiganensis subsp. michiganensis (Cmm) in foreign tomato and chili seed in the agricultural area of Sinaloa, Mexico, besides the growth and cell density of Cmm was evaluated in different selective media under continuous illumination and photoperiod. The results indicate that seeed of 35 varieties of tomatoes was collected; while for Chili seed were 18. This study was supported by farmers (225) which represent 79% of all growers and 32 business engaged in the sale of agro-supplies, provided seeds of varieties and hybrids. Those growers are from six areas (Culiacan, El Tamarindo, Navolato, Culiacan, El dorado and Badiraguato). For detection of Cmm in tomato seed, from 35, only four was variability considering Immunochromatography and ELISA techniques; however, considering chemical and physiological test, the result was negative. Similar results were in 18 varietes of chili seed, where eight showed variability to detect Cmm, and negative by chemical and physiological test. According to the growth and cell density of Cmm, the optimal medium was YDC under pH stable and continuous light conditions. It is recommended to consider the fusion of diagnostic techniques in the emission of a result.
Cowpea plants naturally infected with cowpea mosaic comovirus (CPMV) showed different mosaic, mottle, dwarfing, and vain clearing symptoms. Diseased plants were ollected from certain locations of Alexandria and El-Beheira governorates during the growing seasons from 2011 to 2012. CPMV was detected in infected sap at 8 to 24 days after inoculation by DBIA, indirect ELISA and tissue blot immunoassay (TBIA). Chlorotic local lesions were observed on Chenopodium amaranticolor in infectivity test. By using indirect ELISA and DBIA, CPMV were detected in infected plant sap of serial dilutions up to 1: 400. The incidence of CPMV in 21 day old cowpea seedlings grown from infected seeds was determined by ELISA and positive detection of virus antigen reached 65%. Nitrocellulose membrane and canson paper could be used as solid carriers in TBIA and DBIA for detection of CPMV in infected plant tissues. Results revealed that both faces of nitrocellulose membrane and canson paper could be used as solid carriers in TBIA for detection of CPMV in infected plant tissues. According to reverse transcription polymerase chain reaction (RT-PCR) assay of CPMV infected plant; the amplified product was approximately 800bp of partial coat protein gene. The nucleotide sequences accession number were LN606585 and LN606586. The phylogenetic tree was generated using sequences of CPMV isolates with the other CPMV records from GenBank.
The use of enzyme linked immunosorbent assay (ELISA) for the detection of plant viruses is well documented. It proved to be a very valuable detection tools for the plant viruses. The efficiency of the ELISA technique was for practical purpose independent of the ratio of antibodies to antigen. This avoids the necessity of making specific enzyme conjugates for each antigen to be tested and eliminates the extreme specificity, thus allowing for quantitative evaluation of strain relationships. The advantages of indirect ELISA are sample. It needs only to be macerated and added to the plate. The crude antiserum could be used, although it should be cross absorbed before to prevent spurious host reaction. Single commercially available second antibody conjugate is utilized, thus eliminating the problems of preparing and storing many different conjugated antisera. Blotting technique has become widely used for specific identification of nucleic acid and proteins. This dot assay was modified to detect protein by spotting the antigen on a nitrocellulose membrane and incubating the membrane in test antibody followed by incubation in peroxidase-conjugated second antibody to the first antibody, and by development in 4-chloro-1-naphthol. The above procedure termed dot blot immunobinding assay (DBIA). The technique of tissue blotting on nitrocellulose membrane was described for detection of plant viruses in infected plants. Tissue blots were made by pressing with a firm and gentile force, the freshly cut tissue surface on nitrocellulose membranes. The possibility of using both sides of the nitrocellulose membrane (NCM) by tissue blot immuno assay (TBIA) for the detection plant viruses. In an effort to reduce the cost of virus assays, different types of regular paper were evaluated as possible replacements for the commonly used nitrocellulose membrane (NCM) as the solid phase in the tissue-blot immunoassay (TBIA) were used. Comparisons between different serological methods were demonstrated by many investigators Dot immunobinding was eight times more sensitive for detection of PVX and four times more sensitive for detection of PVS and PVY than DAS-ELISA.
An isolate of zucchini yellow mosaic virus (ZYMV) was obtained from naturally infected squash fruits were grown in Abees region, Alexandria governorate. Disease symptoms were Showing mosaic, yellowing and blistering and absis symptoms. The identification was based on the symptoms developed on diagnostic hosts and serological reactions with antisera to cucumber mosaic cucumovirus (CMV), watermelon mosaic potyvirus 2 (WMV-2) and ZYMV. Squash fruit isolate of ZYMV was transmitted by Aphis gossypii, Aphis neri and Myzus persicae in non-persistent manner. The virus was purified by ultra-centrifugation and PEG. The purified virus had an ultraviolet absorption spectrum typical of a nucleoprotein with A260/280 and A280/260 being 1.1 and 0.91 respectively. The yield of purified virus was 1.62 mg/100g infected leaf tissues. Specific antiserum was prepared and found to have a titer of 1:409600 as determined by indirect ELISA.
Jiawei Zhao, Zhihong Yang, Min He, Qinghua Wang and Renming Hu*
Published on: 19th June, 2018
Although exercise has been proposed to be beneficial to type 2 diabetes, its effects on β-cell function and mass remain unclear. In the present study, the effects of long-term swimming training on the function and mass of β-cells in diabetic OLETF rats were examined. At 44 weeks of age after developing diabetes, the OLETF rats were divided into two groups: a control group and an exercise group. The exercise group had a daily swimming for 12 weeks. While not found with the control rats, in the obese OLETF rats, the exercise reduced the weight gain which was associated with improved glucose tolerance and elevated circulating insulin levels as determined by the oral glucose tolerance test and insulin ELISA. The exercise improved plasma total cholesterol and triglyceride levels, and also significantly increased the islet β-cell mass and pancreatic insulin content associated with decreased β-cell apoptosis and elevated activation of the serine/threonine kinase, Akt. The present studies suggest that exercise improves diabetes symptoms via enhancement of the β-cell mass and function through decreasing glucolipotoxicity and reducing β-cell apoptosis by activating Akt in obese OLETF rats.
Background: Heparin-induced thrombocytopenia/thrombosis (HIT/T) is characterized by a fall in platelet count 5-10days after starting heparin therapy and is diagnosed with specific 4-T clinical features and laboratory tests. This complication is relatively common in Cardiothoracic surgery patients. Objective: To evaluate the positive and negative predictive value of various HIT laboratory tests and assess any correlation between HIT, the underlying diagnosis, underlying procedure, and mechanical cardiac devices. Patients and methods: The patient’s medical records were correlated with two laboratories HIT diagnostic tests, the pan-specific screening test with IgG, IgA, and IgM antibodies, followed by HIT specific IgG ELISA. Results: Total n = 80 patients were assessed, 48% (n = 38) were HIT screen pan-specific negative and 50% (n = 40) were HIT pan-specific positive and 2 cases were inconclusive. 17% (n = 14) were both pan-specific and specific HIT IgG ELISA positive. There were 5 atypical cases. One patient had Eosinophilic myocarditis and was HIT ELISA IgG neg. Argatroban was given on clinical grounds with successful recovery. One patient with Sarcoidosis had an aggressive course and received IV Immunoglobulin (IVIG) but succumbed secondary to liver failure. One patient progressed to gut ischemia and had surgical intervention but succumbed. Two patients with mechanical heart valves were on Argatroban but relapsed and responded to IVIG therapy. Conclusion: Our study indicates that 9/16 (> 50%) HIT-positive patients had valve replacement or cardiac devices suggesting that like knee arthroplasty there is a high incidence of HIT in patients with mechanical heart valves and cardiac devices and this warrants further prospective study.
Background and objectives: YKL-40, a C-reactive protein belongs to the positive acute-phase protein. It is also known as Human chitinase-3-like protein 1(HCI3L1) and is closely related to both acute and chronic inflammation. The present study aimed to detect and estimate the levels of YKL-40 in gingival crevicular fluid (GCF) in patients with healthy periodontium, chronic periodontitis, and rheumatoid arthritis with chronic periodontitis. Materials and methods: Forty-five patients in the age range of 25-55years were included in the study. Patients were divided into three groups: Group I-15 Periodontal healthy patients, Group II-15 Chronic Periodontitis patients, and Group III-15 Rheumatoid arthritis with chronic periodontitis patients. Clinical parameters recorded were Plaque index, Gingival index, Gingival bleeding index, probing depth, and Relative attachment level. GCF samples were analyzed using ELISA. p - value < 0.05 was considered statistically significant. Results: The highest mean YKL-40 concentration in GCF was observed in rheumatoid arthritis with Chronic periodontitis. The mean concentration of YKL-40 in GCF showed a three to four folds increase in its levels when compared to healthy controls (p < 0.001). Contrary, GCF YKL-40 levels between Group II and Group III were not significant.Conclusion: With the increase in severity of periodontal destruction from healthy periodontium to chronic periodontitis, there was a substantial increase in the concentration of YKL-40 in GCF. Correlation of GCF YKL-40 with clinical parameters demonstrated increased severity of the diseases increased its levels
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