To investigate the disturbance in serum levels of interleukin-17 (IL-17) and transforming growth factor-beta1 (TGF-β1) and gene expression of retinoic acid-related orphan receptor-gamma t (ROR-γt) and forkhead box-P3 (FOX-P3) in patients with systemic lupus erythematosus (SLE) and to study their association with disease pathogenicity and activity. Newly diagnosed active patients with SLE (n=88) and healthy volunteers (n=70) were included. Serum IL-17 and TGF-β1 were measured using enzyme-linked immunosorbent assay. Gene-expression profiles of ROR-γt and FOX-P3 were screened using real-time polymerase chain reaction. The IL-17/TGF-β1 and ROR-γt/FOX-P3 levels were also calculated. The mean age of the patients was 30.96±8.25 years; they were 82 women and 6 men. Of the patients, 11.4% manifested mild disease while 88.6% had severe disease. The serum level of TGF-β1 was significantly lower (70.2±34.9 vs. 200.23±124.77 pg/ml), while both IL-17 (614.7±317.5 vs. 279.76±110.65 pg/ml) and IL-17/TGF-β1 (18.5±30.1 vs. 1.66±0.9) levels were significantly higher, in patients than in controls (p<0.0001).The gene-expression level of FOX-P3 (0.6±0.8 vs. 13.68±39.35) was reported to be lower, while ROR-γt (3.9±3.5 vs. 1.99±2.09) and ROR-γt/FOX-P3 (18.6±21.1 vs. 7.63±17.19) levels were significantly higher, in patients than in controls (p<0.0001).

Calcium phosphate nanoparticles (CPNPs) have been synthesized by chemical precipitation method and were characterized by UV-visible spectroscopy (UV-vis), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The antibacterial activity against multi-drug resistant (MDR) gram-negative bacteria Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumonia (K. pneumonia) was performed by well diffusion method, using different concentrations of CPNPs and different combinations of CPNPs with ciprofloxacin (CIP) (CIP-CPNP100, CIP-CPNP50, and CIP-CPNP25). The minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC) were evaluated by the broth dilution method and optical density. Cytotoxicity of nanoparticles was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay on polymorphonuclear cells. Results indicated that synthesized CPNPs sized 28.02 ± 3.2 nm in diameter as average, with distorted spherical shape appears as agglomerates. CPNPs showed no antibacterial activity against MDR bacteria, but combining them with CIP recorded antibacterial activity represented by inhibition zone against MDR bacteria. It was found that the inhibition zone increases when the concentration of CIP and particle size decreases. The MTT assay reveals the acceptable toxicity of the synthesized nanoparticles. The present study can be helpful to formulate nano-drug conjugates as antimicrobial agents in various fields of medical research.

BACKGROUND: Cytokines are inflammatory mediators that regulate multiple processes potentially implicated in inflammation and autoimmunity. AIM: This study aimed to investigate the levels of circulating interleukin (IL)-35, IL-36, and IL-35/IL-36 ratio in Juvenile Idiopathic Arthritis (JIA) patients. METHODS: Fifty-one patients categorized based on the International League of Associations for Rheumatology classification criteria were enrolled, along with 46 healthy controls. Disease activity assessment was conducted according to the Juvenile Arthritis Disease Activity Score (JADAS-27) system. Serum cytokine levels were determined using the sandwich enzyme-linked immunosorbent assay. RESULTS: Compared to controls, the serum levels of IL-36 were significantly higher among JIA children (p = 0.002), and serum IL-35 levels and the IL-35/IL-36 ratio were significantly lower in the patients (p = 0.002, p < 0.001, respectively). Furthermore, the IL-35/IL-36 serum ratio, but not the absolute IL levels, correlated positively with the JADAS-27, giving a good indication of the disease outcome. Further, the IL-35/IL-36 serum ratio showed higher sensitivity and specificity compared to the absolute cytokine levels. CONCLUSION: A significant change in the circulating IL-35 and IL-36 levels and an imbalance in the IL35/IL36 ratio occur in JIA. This study is a novel attempt to explore the role of IL-35 and IL-36 in JIA; further studies are required to support our results. The IL-35/Il-36 ratio is a promising immunological marker to predict, diagnose, and monitor JIA.

The aim: Determine the frequency of anti-viral capsid antigen IgM, IgG and IgA in inflammatory bowel disease. Materials and methods: Case controlled study conducted during involved 60 Crohn’s disease, 60 ulcerative colitis and 60 subjects as a control group with negative gastroin¬testinal symptoms. Diagnosis and disease classification were established according to Montreal disease classification of inflammatory bowel diseases. Measurement of serum anti-VCA IgM, IgG and IgA was done, using ELISA method. Results: The current results showed a higher frequency of EBV seropositivity among both Crohn’s disease and ulcerative colitis 96.67% in comparison with controls 78.33. None statistical significance observed according to sex of patients. IgM were significantly associated with younger than 16 years 33.33%. IgA anti-VCA were significantly frequent within 17-40 years old comprising 100%. Patients with colonic and ileocolonic site of lesions were significantly have frequent anti-VCA IgA 96.43% and 96%. In ulcerative colitis IgM subtype of anti-VCA 35.71% frequent in extensive colitis. Anti-VCA IgG were statistically significant with moderate and severe ulcerative colitis cases 100%. Also, anti-VCA IgA associated with severity of ulcerative colitis 100% of mild cases, 96.43% of moderate cases and 100% of severe cases. Conclusion: EBV seropositivity were detected among IBD cases, however viral infection might be associated with distinct and severe cases that requires anti-viral therapy.

Nanomaterials are increasingly applied to develop new vaccines with new strategies. Implementation of such substances in vaccines will enhance vaccine formula immunogenicity, target delivery, and antigen stability control release. Genetically engineered virus-like particles (VLPs), structurally mimic the viruses and had been successfully used as nano vaccines. VLPs-based vaccines possess the advantage of being safe, effective, and non-infectious. Moreover, due to the optimized nano-size and repetitive structural units of the VLPs, it is suspected that those particles are highly immunogenic, even in absence of adjuvant substances. VLPs could be formulated to carry an array of heterogeneous antigens of different viruses. For all, they are considered as ideal nano vaccine model.

The necessity of nicotine analysis in blood plasma is increasing along with the increased number of smokers and nicotine poisoning cases. One of the analytical methods for nicotine is using high-performance liquid chromatography (HPLC) with ultraviolet (UV) detector because it has been commonly owned by instance in Indonesia. To guarantee accuracy, an analytical method can be used, and it must be validated. This research was the purpose of finding out the validity of the nicotine analysis method in human blood plasma (in vitro) using HPLC with UV detection. Blood plasma samples remained treated with centrifugation procedure by protein denaturation method using acetonitrile. The compounds were analyzed using methanol and buffer acetate 0.01 M (pH 5) 85:15 v/v as a mobile phase on an octadecylsilane column 250 mm, with UV detection at 254 and 260 nm, and flow rate 0.6 mL/minute. Parameter of analytical methods that were validated includes selectivity, accuracy, precision, repeatability, linearity, limit of detection (LoD), limit of quantification (LoQ), and system suitability. According to the result, the selectivity was 2.479, repeatability expressed by its variation coefficient = 0.701%, linearity at range 5–22 μg/mL expressed by coefficient correlation (r) = 0.996. Based on the chromatogram’s area under a curve, the LoD value was found 2.021 μg/mL, LoQ value was 6.737 μg/mL, the accurate percentage was 112.49 to 114.12%, and precision (% CV) was 2.15 to 3.95%. The system suitability from retention time and chromatogram’s area under curve showed % CV 0.70 and 1.64%. According to the experiment result, all parameters meet the requirements of validation criteria.

Bacille Calmette-Guerin (BCG) still the only authenticated vaccine against tuberculosis. Due to its drawbacks, a need for a new formula has emerged. The implication of “Nanovaccinology” is one of the possible alternatives. The non-viral vectors have a low transfection ability. In the context, this work aims to add two adjuvants to a calcium phosphate nanoparticles (CPNPs) functionalized with early secreted antigenic target 6-kilo dalton ( ESAT-6) cloned pcDNA3.1(+) plasmid. ESAT-6 gene is specific to mycobacterium tuberculosis complex (MTC) and encodes a T-cell antigen. The adjuvants in practice are Herring protamine and cytosine-phosphodiester bond-guanosine-oligodeoxynucleotide 7909 ( CpG-ODN 7909). Each has a different strategy in enhancing immune response; protamine is particulate adjuvant while CpG is an immunopotentiator substance. Nano complex was transfected into THP-1 monocytic cell line after its activation to a macrophage via 100nM PMA. Cellular immune response, interleukin-12 (IL-12), and tumor necrosis factor –alfa (TNF-ɑ) also ESAT-6 protein production were assayed via the Sandwich ELISA technique. Results revealed that CPNPs offer only partial protection to the adsorbed plasmid against enzymatic degradation. Nano complex formula with two adjuvants resulted in significantly higher cellular immune response comparing to formula carrying one adjuvant. In conclusion, the implication of CPNPs in gene delivery accompanied with two adjuvants each possess different strategy, will result in partial protection to the delivered gene with upsurge cellular immune response.

Autophagy is a primordial form of eukaryotic innate immunity which arises as a cellular response to stress conditions such as growth factors withdrawal, mitochondrial and other organelles damage, nutrients deficiency and inflammation and assists in removal of damaged organelles. This study aims to investigate the role of autophagic flux (LC3ІІ/І) in rheumatoid arthritis in terms of disease severity represented by DAS-28 and immunological markers represented by TNF-ɑ and NLRP-3 also to study the possible effect of certain rheumatoid arithritis drugs (MTX and infliximab) on autophagic flux. The present study involves sixty rheumatoid arthritis patients, thirty of them receiving chemical drug (MTX) and other thirty receiving biological treatment (Infliximab), comparing results to healthy control group consist of thirty people. These two groups of rheumatoid arthritis patient were subjected to the molecular measurement of the autophagic flux level presented by LC3II/I ratio and NLRP3 level by (rtPCR) and serum TNF-α level by indirect ELISA. Autophagic flux presented by LC3ІІ /LC3І ratio, showed a highly significant correlation with the DAS-28, NLRP-3 and TNF-ɑ level,0.577 and 0.855, 0.640 respectively, P value ˂ 0.001.The autophagic flux in the MTX patients was 2.81 comparing to 1.91 in the Infliximab group,P value is ˃ 0.001 pointing to the absence of any significant difference between them. This study referred to the presence of a detectable correlation between autophagy and rheumatoid arthritis severity presented by both clinical and immunological aspects, on the other hand both of MTX and Infliximab seem to affect autophagy in a similar strength