Pre-Clinical Evaluation of the Nanoliposomal antiPCSK9 Vaccine in Healthy Non-Human Primates

Background: Our previous studies showed the safe preventive and therapeutic effects of immunization using the nanoliposomal antiPCSK9 vaccine called “Liposomal Immunogenic Fused PCSK9-Tetanus plus Alum adjuvant” (L-IFPTA), in mouse models of atherosclerosis. Here we aimed to ascertain the immunogenicity and safety of the L-IFPTA vaccine in a pre-clinical study in healthy non-human primates. Methods: Five male rhesus macaque monkeys were subcutaneously immunized with the L-IFPTA vaccine, four times with bi-weekly intervals. To evaluate immunogenicity, the plasma antiPCSK9 antibody in immunized monkeys was detected and quantified using the ELISA method. The functionality of the induced antiPCSK9 antibodies was determined by the PCSK9/LDLR in vitro binding assay kit. The safety of the vaccine was tested using the evaluation of several major circulating indicators including plasma lipid alterations, inflammatory biomarkers and organ injury biomarkers. Results: The resultant data indicated that the L-IFPTA vaccine significantly and highly induced the generation of functional and safe antiPCSK9 antibodies in immunized monkeys. Plasma levels of specific biomarkers indicating organ performance including creatinine, urea, uric acid, bilirubin, ALP, AS, ALT and TSH were not significantly altered. After immunization in healthy monkeys, non-prespecified endpoints (plasma levels of TC, LDL-C, VLDL-C and TG) were non-significantly reduced by 11.6 ± 36%; 16 ± 28%; 22 ± 53% and 24 ± 51%, respectively, while HDL-C was slightly increased by 2 ± 64%. There were also no significant changes in plasma levels of pro- and anti-inflammatory biomarkers. Conclusion: The L-IFPTA vaccine could efficiently stimulate the host humoral immune response to produce active antibodies that inhibit plasma PCSK9 while not provoking systemic inflammation and not adversely affecting organ performance.

PROTESTANT CHORALE AND METHODS OF ITS PROCESSING

The article is devoted to the consideration of the Protestant chorale, the preconditions of its origin and the first examples. The situation from the history of the early Lutheran church is described, when Martin Luther introduced communal choral singing. The main task set in the work is to analyze the main methods of processing Protestant chorales in the works of prominent Western European and domestic composers. Several vocal, instrumental and vocal-instrumental genres are considered, which are based on choral. It is noted that the most common means of processing the chorale is its harmonization, designed for both choral and organ performance. In the Renaissance and Baroque, the practice of creating figures on the choral became quite common, as well as the use of the choral as the theme of the fugue – fugue choral. Another way of elaborating the chorale was the technique of cantus firmus, while the theme of the chorale was located in the lower voice or in the tenor in rhythmic expanse, forming the basis of the whole composition. One of the leading genres of organ music – choral preludes – is considered, special attention is paid to the works of JS Bach. The principle of construction of choral fantasies is analyzed separately. Among other genres, parties, toccatas, masses, oratorios, and other works were created on the basis of chorales. In conclusion, there is a modern tendency to return to the practice of using chant in the liturgy in its original form, as they sounded earlier.

POLA INFEKSI Streptococcus agalactiae STRAIN NP105O DAN N14G PADA IKAN NILA (Oreochromis niloticus)

Infeksi Streptococcosis yang disebabkan oleh bakteri patogen Streptococcus agalactiae dengan karakteristik strain berbeda menjadi permasalahan utama pada budidaya ikan nila. Penelitian ini bertujuan untuk menelaah pola infeksi bakteri S. agalactiae strain NP105O dan N14G, melalui performa organ target, gejala klinis, serta hematologi ikan nila Oreochromis niloticus. Karakterisasi S. agalactiae berdasarkan pada SNI dan API 20 STREP, uji pertumbuhan bakteri dilakukan dengan metode total plate count (TPC). Pengujian eksistensi bakteri dengan cara menginjeksikan S. agalactiae secara intraperitoneal (IP) dengan konsentrasi 107 CFU.mL-1, dan pengamatan dilakukan dengan pengambilan darah dan dikultur di media BHIA. Hasil uji biokimia dan konfirmasi menggunakan API 20 STREP menunjukkan bahwa isolat terkonfirmasi positif sebagai S. agalactiae, dan NP105O dideteksi sebagai bakteri â-hemolitik. Pertumbuhan bakteri NP105O lebih cepat daripada N14G, namun eksistensi di darah masing-masing selama 72 jam dan 24 jam. Hasil pengamatan performa darah menunjukkan bahwa glukosa dan leukosit mengalami peningkatan signifikan masing-masing 53,5 ± 2,12 mg.dL-1 dan 6,51 ± 0,89 (105 sel.mm-3), sedangkan hematokrit dan eritrosit mengalami penurunan signifikan (P<0.05) masing-masing 21,10 ± 0,07% dan 14 ± 4,5 (105 sel.mm-3) pascainjeksi S. agalactiae. Gejala klinis pascainfeksi berupa melanosis, respons lambat, anorexia, ocular opacity, purulens, unilateral atau bilateral eksoptalmia, gasping, erratic, C-shape, dan whirling. Pola infeksi S. Agalactiae strain NP105O dan N14G berbeda pada ikan nila, dan sangat dipengaruhi oleh keberadaan bakteri pada organ ginjal, otak, dan mata.Streptococcosis infection caused by different strains of pathogenic bacteria Streptococcus agalactiae has been a major problem in tilapia culture. This research aimed to examine the patterns of S. agalactiae infection of NP105O and N14G strains, through targeted organ performance, clinical symptoms and hematological signs of tilapia, Oreochromis niloticus. The characterization of S. agalactiae was based on SNI and API 20 STREP. Bacterial growth test was carried out using the total plate count (TPC) method. Bacterial infection was performed by injecting S. agalactiae intraperitoneally (IP) with a concentration of 107 CFU.mL-1, and the observations were carried out by extracting and culturing the fish blood in BHIA media. The result of the biochemical test and API 20 STREP confirmed that the isolates was identified as S. agalactiae and NP105O strain was detected as â-hemolytic bacteria. The growth of NP105O strain was faster than N14G strain, where their clear presence in blood was observed at 72 and 24 hours, respectively. The result of hematological parameters showed that glucose and leukocytes increased significantly with values of 53.5 ± 2.12 mg.dL-1 and 6.51 ± 0.89 (105 cell.mm-3), respectively. On the other hand, hematocrit and erythrocytes decreased significantly (P<0.05) 21.10 ± 0.07% and 14 ± 4.5 (105 cell.mm-3) post-S. agalactiae injection. Clinical signs post-infection consisted of melanosis, slow response, anorexia, ocular opacity, purulence, unilateral or bilateral exophthalmos, gasping, erratic movement, C-shape and whirling. NP105O, and N14G strains show different patterns of infections on tilapia and strongly influenced by the presence of bacteria in the kidneys, brain, and eyes.