The analysis of anticcp antibodies in the serum: a comparison between the patients with rheumatoid arthritis

Rheumatoid Arthritis (RA) is a chronic systemic autoimmune disease that causes inflammation, pain, stiffness and destructive changes in the joints. Although, Rheumatoid Factor (RF), has been the primary blood test used to detect RA, the anti-ccp antibodies detection test is a relatively new assay to detect the citrulline antibodies in blood. These autoantibodies are produced by immune system in response to a perceived threat of citrulline, an amino acid produced from arginine in the citrullination process. The objective of this study was to investigate the presence and prediction value of anti-ccp in RA patients and evaluate its sensitivity and specificity comparing to that of classic laboratory tests, CRP and RF. The serum of 84 patients with RA and 80 healthy control subjects were enrolled into the study. The anti-ccp, RF and CRP levels in the serums were assayed by ELISA and agglutination procedure, respectively. Our results provided evidence that anti-ccp level was significantly higher in patients with RA comparing to that of corresponding controls (p<0.0001). Anti-ccp was found to have the highest sensitivity and specificity (91%-91%) comparing to the other two tests (RF, CRP). The latter tests were found to have (97%- 92%) and (27%- 75%) sensitivity and specificity, respectively. The diagnostic value of anti-ccp is better than RF and CRP, individually. It can be detected early in the disease in unselected early arthritis patients. It is recommended to use RF test together with anti-ccp antibodies detection, in RA patients to ensure a higher diagnostic effectiveness

Molecular detection, virulence genes, biofilm formation, and antibiotic resistance of salmonella enterica serotype enteritidis isolated from poultry and clinical samples

Background: Salmonella spp. is one of the most important zoonotic pathogens transmitting among human and animals. Due to the similarity of antibiotic classes used to treat animals and humans, there is a high risk for emerging the multi-drug resistant (MDR) strains. Objectives: The current study aimed at evaluating molecular detection, virulence genes, biofilm formation, and antibiotic resistance of Salmonella enterica serotype enteritidis recovered from poultry and clinical isolates. Methods: A total of 282 isolates were recovered from chicken meat, live poultry feces, eggs, and human feces in Iran. The presence of virulent factors in the isolates was confirmed using biochemical and microbiological tests. The presence of Salmonella genus was determined using antiserum. Triplex polymerase chain reaction (PCR) was performed to detect Salmonella spp., serogroup D and the discriminate S. enteritidis from other species. Kirby-Bauer disk diffusion method was applied to perform the susceptibility testing. Quantification of biofilm formation was determined in 96-well microtiter plates as recommended by the defined protocol. The data were then analyzed with SPSS using consensus tables and Chi-square test. Results: Based on the results, all the isolates were positive for invA, sdiA, hilA, and ratA. Moreover, spvC had the lowest prevalence (37.6). Of all strains, 67 were MDR, 51.7 of which were recovered from humans. Furthermore, 34.5 of isolates were strong biofilm producers. There was a significant correlation between the strong biofilm formation and the antibiotic resistance to colistin, ceftazidime, chloramphenicol, gentamicin, trimethoprim, penicillin, and trimethoprim-sulfamethoxazole. Conclusions: The results of the current study showed a significant correlation between the strong biofilm formation and the antibiotic resistance to some antibiotics. © 2018, Author(s)

A Portable RT-LAMP/CRISPR Machine for Rapid COVID-19 Screening.

The COVID-19 pandemic has changed people's lives and has brought society to a sudden standstill, with lockdowns and social distancing as the preferred preventative measures. To lift these measurements and reduce society's burden, developing an easy-to-use, rapid, and portable system to detect SARS-CoV-2 is mandatory. To this end, we developed a portable and semi-automated device for SARS-CoV-2 detection based on reverse transcription loop-mediated isothermal amplification followed by a CRISPR/Cas12a reaction. The device contains a heater element mounted on a printed circuit board, a cooler fan, a proportional integral derivative controller to control the temperature, and designated areas for 0.2 mL Eppendorf® PCR tubes. Our system has a limit of detection of 35 copies of the virus per microliter, which is significant and has the capability of being used in crisis centers, mobile laboratories, remote locations, or airports to diagnose individuals infected with SARS-CoV-2. We believe the current methodology that we have implemented in this article is beneficial for the early screening of infectious diseases, in which fast screening with high accuracy is necessary

The evolving landscape of predictive biomarkers in immuno-oncology with a focus on spatial technologies.

Immunotherapies have shown long-lasting and unparalleled responses for cancer patients compared to conventional therapy. However, they seem to only be effective in a subset of patients. Therefore, it has become evident that a greater understanding of the tumor microenvironment (TME) is required to understand the nuances which may be at play for a favorable outcome to therapy. The immune contexture of the TME is an important factor in dictating how well a tumor may respond to immune checkpoint inhibitors. While traditional immunohistochemistry techniques allow for the profiling of cells in the tumor, this is often lost when tumors are analysed using bulk tissue genomic approaches. Moreover, the actual cellular proportions, cellular heterogeneity and deeper spatial distribution are lacking in characterisation. Advances in tissue interrogation technologies have given rise to spatially resolved characterisation of the TME. This review aims to provide an overview of the current methodologies that are used to profile the TME, which may provide insights into the immunopathology associated with a favorable outcome to immunotherapy

Adenosine Deaminase 1 as a Biomarker for Diagnosis and Monitoring of Patients with Acute Lymphoblastic Leukemia

Background: Acute lymphoblastic leukemia (ALL) is known as the most prevalent pediatric malignancy all around the world. Identification of specific biomarker is necessary for early diagnosis and effective therapy. It is believed that Adenosine deaminase (ADA) as an enzyme involved in the purine salvage pathway increases in ALL patients. Herein, the quantity and pattern of ADA isoenzymes were surveyed among ALL patients in comparison to healthy subjects. Methods: Serum and RBC samples of three different groups of ALL patients, including newly diagnosed cases without any drugs administration, subjects with the relapsed disease, patients in the remission stage after therapy, and the healthy subjects were enrolled in the study. Then, the activity and pattern of ADA1, ADA2 and ADA1+cp were determined using ADA kit and electrophoresis on SDS-PAGE, respectively. To confirm the presence of ADA enzyme, the fresh serums, extractions from erythrocytes, JM cell line as a human T lymphocyte line and J774 A.1 as mouse monocyte line were electrophoresed on 1.2 agarose gel and stained with the specific dye. Results: The activities of ADA1 isoenzyme and total ADA in new cases and subjects with the relapsed disease were significantly higher than their activities in the patients in the remission stage and healthy controls (p<0.001). The unbounded ADA1 isoenzyme was found to exist in the erythrocyte, lymphocyte and monocyte. But in serum, all the ADA1 was bounded to the cp protein. Conclusions: ADA1 is the key isoenzyme elevating in ALL patients, therefore this isoenzyme could be a useful biomarker to diagnose ALL patients and monitor their therapies. © 2017 Mina Ebrahimi-Rad et al

Adenosine Deaminase 1 as a Biomarker for Diagnosis and Monitoring of Patients with Acute Lymphoblastic Leukemia

Background: Acute lymphoblastic leukemia (ALL) is known as the most prevalent pediatric malignancy all around the world. Identification of specific biomarker is necessary for early diagnosis and effective therapy. It is believed that Adenosine deaminase (ADA) as an enzyme involved in the purine salvage pathway increases in ALL patients. Herein, the quantity and pattern of ADA isoenzymes were surveyed among ALL patients in comparison to healthy subjects. Methods: Serum and RBC samples of three different groups of ALL patients, including newly diagnosed cases without any drugs administration, subjects with the relapsed disease, patients in the remission stage after therapy, and the healthy subjects were enrolled in the study. Then, the activity and pattern of ADA1, ADA2 and ADA1+cp were determined using ADA kit and electrophoresis on SDS-PAGE, respectively. To confirm the presence of ADA enzyme, the fresh serums, extractions from erythrocytes, JM cell line as a human T lymphocyte line and J774 A.1 as mouse monocyte line were electrophoresed on 1.2 agarose gel and stained with the specific dye. Results: The activities of ADA1 isoenzyme and total ADA in new cases and subjects with the relapsed disease were significantly higher than their activities in the patients in the remission stage and healthy controls (p<0.001). The unbounded ADA1 isoenzyme was found to exist in the erythrocyte, lymphocyte and monocyte. But in serum, all the ADA1 was bounded to the cp protein. Conclusions: ADA1 is the key isoenzyme elevating in ALL patients, therefore this isoenzyme could be a useful biomarker to diagnose ALL patients and monitor their therapies. © 2017 Mina Ebrahimi-Rad et al

Evaluation of serum adenosine deaminase and its isoenzymes in patients with ovarian cancer

Ovarian cancer is the most lethal gynecological cancer worldwide. There are great relationships between the activities of adenosine deaminase (ADA), one of the enzymes in purine nucleotide pathway and carcinogenic process. In the present study the activities of the total ADA, ADA1 and ADA2 were measured in the sera of the patients with ovarian cancer. In this study, activities of tADA, ADA1 and ADA2 were assessed in sera of 30 patients with ovarian cancer and 30 normal control individuals, using a modified Ellis method in which only ADA2 activity was measured in the present of a specific inhibitor, erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA). Our results showed that the tADA, ADA1, and ADA2 serum activities of patients were found to be significantly increased (P < 0.05) than those of healthy control group. Although, ADA and its isoenzymes were not the specific markers for diagnosis of ovarian cancer, measurement of their activities may be used as a diagnostic means in ovarian cancer as well as the other analytical procedures