WITNESS PROTECTION PROGRAMS FOR JUSTICE COLLABORATORS – COMPARATIVE OVERVIEW: POSITIVE LEGAL SOLUTIONS IN THE REPUBLIC OF SERBIA, THE REPUBLIC OF NORTH MACEDONIA, USA, ENGLAND AND ITALY

Justice collaborators are exposed to the highest risk of retaliation, as they have decided to testify against their former associates (members and leaders of criminal organizations) through cooperation with prosecution authorities in order to gain certain privileges. In one person, the justice collaborator, two procedural features are merged: the collaborator’s during the conclusion of the agreement, and the witness’, after the agreement was accepted by the court and his testimony given. After the testimony was given and the criminal proceeding was completed, these persons are not unattended, but every state that faces threats of organized forms of criminal activity is obliged to provide them both procedural and non-procedural (Program) protection. This paper presents the procedure of providing program protection to justice collaborators in the legislations of the Republic of Serbia, the Republic of North Macedonia, the United States of America, England and Italy, the extent of its efficient implementation and the real and potential difficulties in front of persons included in the Program

Molecular genetic characterization of Prototheca zopfii as causative agent of mastitis in cows in Serbia

Jednoćelijske alge Prototheca sp. su mikroorganizmi značajni za veterinarsku i humanu medicinu jer više vrsta navedenog roda ima patogeni potencijal i, koliko je danas poznato, to su jedine alge koje izazivaju infekcije životinja i ljudi. Mastitis goveda izazvan prototekama je najčešći oblik prototekoze među životinjama javlja se u vidu hroničnog subkliničkog ili umerenog kliničkog mastitisa. Prototekalni mastitis goveda u najvećem broju slučajeva izaziva vrsta P. zopfii i to genotip 2, a znatno ređe vrsta P. blaschkeae. Na teritoriji Srbije zabeleženi su slučajevi mastitisa rezistentnih na antibiotski tretman u kojima je kao uzročnik potvrđena alga P. zopfii, ali su dosadašnja istraživanja bila ograničenog obima i nisu obuhvatila molekularnogenetičku karakterizaciju uzročnika, tako da nije poznato koji je genotip P. zopfii bio u pitanju. Zbog toga je bilo opravdano da se u ovoj tezi ispita prevalencija mastitisa izazvanih sa P. zopfii na nekim ispitivanim farmam Srbiji i izvrši molekularnogenetička karakterizacija P. zopfii izolovanih u slučajevima kliničkog i subkliničkog mastitisa. Od ukupno 494 uzorka mleka krava obolelih od mastitisa izazvanim algom iz roda Prototheca, prototeka je izolovana iz 39 uzoraka, od toga 32 uzorka su iz mleka krava koje su obolele od subkliničkih mastitisa i 7 uzoraka je iz mleka krava sa znacima kliničkog mastitisa. 4. Ukupno izolovanih algi iz roda Prototheca bilo je 39, od toga Prototheca zopfii genotip 2 identifikovana je u 37 (94,8%) izolata i samo dva izolata su potvrđena kao (5,2%) Prototheca zopfii genotip 1. Ukupno 25 (64,11%) izolata P. zopfii je bilo osetljivo na nistatin, 16 (41,03%) izolata na streptomicin, 9 (23,07%) na kanamicin i 6 (15,38%) na gentamicin, na osnovu čega se nistatin i aminoglikozidni preparati mogu preporučiti u terapiji prototekalnog mastitisa u Republici Srbiji.Onecelled algae Prototheca sp. are Microorganisms important for veterinary and human medicine, because more than one species of said genus has pathogenic potential and how much is known today, these are the only algae that cause infections in animals and humans. Mastitis in cattle caused by Prototheca sp. a is the most common form of protothecosis among animals and it occurs in the form of chronic subclinical or moderate clinical mastitis. Protothecal bovine mastitis in most cases causes type P. zopfii, genotype 2, and much rarer type P. blaschkeae. In Serbia, there were cases of mastitis resistant to an antibiotic that has been caused by the algae P. zopfii, but previous studies were limited in scope and are not included molecular genetic characterization of the pathogen, so it is not known which genotype P. zopfii was concerned. Therefore it was justified that in this thesis we examine the prevalence of mastitis caused by P. zopfii tested on some farms in Serbia and to perform molecular genetic characterization of P. zopfii isolated in cases of clinical and subclinical mastitis. From a total of 494 samples of milk Prototheca sp. was isolated from 39 samples, of which 32 samples were from the milk of cows that are suffering from subclinical mastitis and 7 samples from the milk of cows with signs of clinical mastitis. Total isolated Prototheca sp. was 39, of which was identified 37 (94.8%) Prototheca zopfii genotype 2 and only two samples (5.2%) Prototheca zopfii genotype 1. A total of 25 (64.11%) of the P. zopfii isolates was sensitive to nystatin, 16 (41.03%) isolates was sensitive to streptomycin, 9 (23.07%) and kanamycin, and 6 (15.38%) was sensitive to gentamicin, based on these findings the aminoglycoside and nystatin preparations can be recommend for the treatment of protothecal mastitis in the Republic of Serbia

Makrakantarhinhoza svinja

Macracantarhynhosis are swine parasitoses caused by hookworm Macracanthorhynchus hirudinaceus. Development of parasites included intermediate host in genus Scarabidae (Melontha vulgaris, Cetonia aurata, Polyphilla fullo, Anomalia vitis i dr.). Infection occurred when swine eat infected insects and usually occurred at extensive breeding. Adult hookworm are 10-17 cm long and 4-10 mm wide. They live in small intestine of swine, some carnivores and human. Pathology changes were at deep part of intestinal mucus where we found deep wund and bacterial infection at fixation places. Clinical signs are apathy.Makrakantarhinhoza je parazitoza uzrokovana sa akantocefalom Macracanthorhynchus hirudinaceus. Razvoj parazita ide preko prelaznog domaćina - koleoptera iz roda Scarabidae (Melontha vulgaris, Cetonia aurata, Polyphilla fullo, Anomalia vitis i dr.). Infekcija nastaje kada svinje pojedu zaražene koleoptere, tako da se infekcije sreću u ekstenzivnom držanju. Paraziti su dugi 10-17 cm i široki 4-10 mm. Parazitiraju u tankim crevima svinja. Patološko delovanje se ispoljava u vidu dubokih rana u sluznici creva, toksičnim delovanjem metabolita i posledičnim bakterijskim infekcijama na mestu fiksacije. Od kliničkih simptoma prisutni su apatija, mršavljenje, anemija, konvulzija, a kod jakih infekcija i uginuća.

Istraživanje polimorfizma gena za κ-kazein i beta-laktoglobulin u buše i holštajnfrizijske pasmine mliječnih krava u Srbiji

The aim of this study was to determine the distribution of κ-casein (κ-CN) and β-lactoglobulin (β-Lg) genotypes in the autochtonous (Busha) and dairy (Holstein-Friesian, HF) cattle breeds with PCR-RFLP (polymerase chain reaction - restriction fragment length polymorphism). For the amplification of κ-CN and β-Lg gene fragments specific primers were used. After digestion with specific endonucleases genotypes were determined for both genes in 18 Busha and 19 HF cows. The results showed that κ-CN gene was represented with the AA genotype in 31.58 % HF cows, AB in 52.63 % cows, whilst the genotype BB was found in 15.79 % cows only. Among the examined Busha cattle 44.44 % cows had AA genotype and 55.56 % genotype AB for κ CN. As for β-Lg gene in HF breed, AA genotype was found in 26.31 % cows, AB in 63.16 % and BB in 10.53 % cows. In Busha cows the following genotypes were established for β-Lg gene: AA in 44.44 % cows and AB in 55.56 %, whilst BB genotype was not found. These results indicate that Busha cows had a higher presence of A allelic forms of both genes (k-CN and β-lactoglobulin) than HF cows.Cilj istraživanja bio je utvrditi raspodjelu genotipova kapa-kazeina (κ-CN) i beta-laktoglobulina (β-Lg) u autohtonih (buša) i mliječnih (holstein, HF) pasmina goveda primjenom PCR-RFLP. Za amplifikaciju κ-CN i β-Lg fragmenata gena korištene su specifične oligonukleotidne početnice. Nakon digestije posebnim endonukleazama (Hinf I i Hae III) genotipovi su određeni za oba gena u 18 buša i 19 HF krava. Rezultati su pokazali da je κ-CN gen utvrđen genotipom AA u 31,58 % HF krava, AB u 52,63 % krava, dok je genotip BB utvrđen u samo 15,79 % krava. Od krava pasmine buša 44,44 % je imalo AA genotip i 55,56 % genotip AB za κ-CN. Što se tiče β-Lg gena u HF pasmine, AA genotip pronađen je u 26,31 % krava, AB u 63,16 % i BB u 10,53 % krava. U krava pasmine buša sljedeći genotipovi su utvrđeni za β-Lg gen: AA u 44,44 % i AB u 55,56 % krava, dok BB genotip nije utvrđen. Ovi rezultati pokazuju da je u krava pasmine buša veća prisutnost A alelne forme za oba ispitivana gena (za k-CN i β-laktoglobulin) nego kod HF krava

Development of immuno-PCR for sensitive quantification of SARS-CoV-2 nucleocapsid protein

Accurately diagnosing people with suspected SARS-CoV-2 infection is essential to help manage COVID-19. Currently available SARS-CoV-2 diagnostics detect either RNA of the virus by RT-PCR or the presence of viral antigens in biological fluids by ELISA or similar techniques. Low sensitivity of antigen tests could lead to the risk of false negative results. Therefore, this study aimed to develop a highly sensitive immuno-PCR method for quantifying SARS-CoV-2 nucleocapsid (N) protein that combines the specificity of sandwich ELISA with the sensitivity of PCR. Recombinant N protein fragment was produced in E. coli as an expression system and purified using immobilized metal ion affinity chromatography. The antibodies against the N protein were raised in rabbits and mice. High-affinity polyclonal mice and rabbit N protein-specific antisera were purified using ammonium sulfate precipitation and used to develop sandwich ELISA for the quantification of N protein. Mice polyclonal serum was used as a capture for N protein. N protein bound to mice antibodies was detected with rabbit polyclonal sera. A double-stranded amino-DNA molecule of 77 base pairs was PCR-synthesized, covalently conjugated to a secondary goat anti-rabbit antibody and subsequently amplified and quantified by real-time PCR. The results were compared to analogous sandwich ELISA consisting of alkaline phosphatase-labeled goat anti-rabbit antibody. The sensitivity of immuno-PCR for quantification of N protein was increased by up to 7-fold compared to analogous ELISA, having a limit of detection of 92 pg/mL and a limit of quantification of 840 pg/mL. The developed immuno-PCR method thus has the potential to be used as a new antigen test for COVID-19 and beyond

Molekul oštećenja bubrega-1 (KIM-1) i akvaporin-1 (AQP-1) u urinu kod bolesnika sa karcinomom svetlih ćelija bubrega - potencijalni neiznvazivni biomarkeri

Background/Aim. Kidney injury molecule-1 (KIM-1) and aquaporin-1 (AQP-1) are potential early urinary biomarkers of clear renal cell carcinoma (cRCC). The aim of this study was to ascertain relationship between the urine concentrations KIM-1 and AQP-1 with tumor size, grade, pT stage and type of operation (radical or partial nephrectomy) in patients with cRCC. Methods. Urinary concentrations of urinary KIM-1 (uKIM-1) and urinary AQP-1 (uAQP-1) were determined by commercially available ELISA kits. The analysis included 40 patients undergoing partial or radical nephrectomy for cRCC and 40 age- and sex-matched healthy adult volunteers. Results. The median preoperative concentrations of KIM-1 in the cRCC group [0.724 ± 1.120 ng/mg urinary creatinine (Ucr)] were significantly greater compared with controls (healthy volunteers) (0.210 ± 0.082 ng/mgUcr) (p = 0.0227). Postoperatively, uKIM-1 concentration decreased significantly to control values (0.177 ± 0.099 ng/mgUcr vs 0.210 ± 0.082 ng/mgUcr, respectively). The size, grade and stage of tumor were correlated positively with preoperative uKIM-1 concentrations. Contrary to these results, concentrations of uAQP-1 in the cRCC group were significantly lower (0.111 ± 0.092 ng/mgUcr) compared with the control group (0.202 ± 0.078 ng/mgUcr) (p = 0.0014). Postoperatively, the concentrations of uAQP-1 increased progressively up to control values, approximately. We find no significant correlation between preoperative uAQP-1 concentrations and tumor size, grade and stage. Conclusion. uKIM-1 was found to be a reliable diagnostic marker of cRCC, based on its significantly increased values before and decreased values after the nephrectomy.Uvod/Cilj. Molekul oštećenja bubrega-1 (KIM-1) i akvaporin-1 (AQP-1) su potencijalni rani biomarkeri karcinoma svetlih ćelija (cRCC). Cilj ove studije bio je da se utvrdi povezanost između koncentracija KIM-1 i AQP-1 u urinu i veličine, gradusa, stadijuma i vrste operacije (radikalna ili parcijalna nefrektomija) kod bolesnika sa cRCC. Metode. Urinarne koncentracije KIM-1 i AQP-1 određene su primenom komercijalnih ELISA kitova. Analizom je bilo obuhvaćeno 40 bolesnika koji su bili podvrgnuti parcijalnoj ili radikalnoj nefrektomiji zbog tumora bubrega i 40 zdravih odraslih ispitanika. Grupe su bile komparabilne po polu i godinama života. Rezultati. Srednja preoperativna koncentracija urinarnog KIM-1 (uKIM-1) u cRCC grupi [0,724 ± 1,120 ng/mg kreatinia u urinu (Ucr)] bila je statistički značajno viša u poređenju sa koncentracijom u kontrolnoj grupi (0,210 ± 0,082 ng/mgUcr ) (p = 0,0227). Postoperativno, koncentracija uKIM-1 značajno je padala i približavala se vrednosti u kontrolnoj grupi (0,177 ± 0,099 ng/mgUcr nasuprot 0,210 ± 0,082 ng/mgUcr). Veličina, gradus i stadijum tumora bili su u pozitivnoj korelaciji sa preoperativnim koncentracijama uKIM-1. Nasuprot ovim rezultatima, koncentracija urinarnog AQP-1 (uAQP-1) u cRCC grupi bila je značajno niža (0,111 ± 0,092 ng/mgUcr) u poređenju sa kontrolnom grupom zdravih osoba (0,202 ± 0,078 ng/mgUcr) (p = 0,0014). Postoperativno, koncentracija uAQP-1 progresivno se povećavala, približno do vrednosti u kontrolnoj grupi. Nismo našli značajnu korelaciju između preoperativnih koncentracija uAQP-1 i veličine, gradusa i stadijuma tumora. Zaključak. uKIM-1 bi mogao biti dodatni pouzdani dijagnostički marker za cRRC na osnovu njegove značajno više preoperativne koncentracije i sniženja vrednosti nakon nefrektomije

Novi metodološki pristup integrisanom upravljanju održivim urbanim razvojem u Republici Srbiji (na primeru Strategije održivog i integralnog urbanog razvoja Republike Srbije do 2030.godine)

Tehničko rešenje ispunjava uslove za dodelu kategorije M82 - novo tehničko rešenje (metoda) primenjena na nacionalnom nivo

Стратегија одрживог урбаног развоја Републике Србије до 2030. године [награда 28. Међународни салон урбанизма]

Влада Републике Србије је 13.јуна 2019.године усвојила националну политику урбаног развоја. Стратегија представља кохерентан сет одлука, вођених од стране националне владе кроз процес сарадње различитих актера у формулисању заједничке визије и циљева, којима се усмерава дугорочни трансформативни, продуктивни, инклузивни и резилијентан урбани развој. Ослања се на интегрални и координисани приступ решавању кључних и ургентних питања урбаног развоја, како би се побољшао квалитет живота унапређењем коришћења територијалног/урбаног капитала и креативних ресурса, и подстицањем раста (иновативне) урбане економије и ефикасноог управљања урбаним развојем. Посебан допринос стретегије је постигнути политички консензус у дефинисању 6 приоритетних националних програма урбаног развоја. Посред традиционалних, стратегија уводи и иновативне управљачке инструменте, који су од значаја за европске интеграције Србије; они ће омогућити доступност финансирања урбаног развоја из различитих извора, укључујући националне, ЕУ и глобалне фондове

Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species

In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests

EVAPOTRANSPIRATION DEFINING

Za zadano područje Požega u razdoblju od 2001. do 2010.godine, koristeći službene podatke i stručnu literaturu, potrebno je izračunati moguću evapotranspiraciju pomoću odabrane dvije metode, te dobivene rezultate međusobno usporediti. Moguća evapotranspiracija proračunata je pomoću Thornthwaiteove i Blaney-Criddleove metode. U proračunu se određuju potrebni koeficijentni i faktori koji utječu na moguću evapotranspiraciju i koriste se u jednadžbama za proračun. Usporedbom dviju metoda utvrđeno je da se godišnje moguće evapotraspiracije razlikuju zbog različitih parametara koji se koriste u jednadžbama proračuna