Kultivisanje, održavanje i primena Caenorhabditis elegans u biomedicinskim istraživanjima

Caenorhabditis elegans je neparazitska nematoda koja živi u zemljištu. Prvi podaci o primeni C. elegans u eksperimentalne svrhe potiču još iz 1965. kada je Sidni Brener, južnoafrički biolog, proučavao ponašanje i razvoj životinja (naročito neuralni razvoj) na ovoj nematodi. Nedugo zatim C. elegans je postao jedan od napopularnijih eksperimentalnih modela u biomedoicinskim istraživanjima. Tome u prilog govori činjenica da su do danas tri Nobelove nagrade dodeljene naučnicima koji su istraživanja upravo radili na C. elegans, a pored toga, dobijena su i mnoga druga značajna otkrića u različitim oblastima biomedicine. Veoma brz i kratak životni ciklus (3-5 dana), velikog broja potomaka i lako kultivisanja u laboratorijskim uslovima neke su od glavnih prednosti C. elegans. Za kultivaciju ove nematode koristi se podloga za rast nematoda (Nematode Growth Median) u koju se dodaje E. coli OP 50 kojom se C. elegans hrani. Postoje dva pola: samooplodni hermafroditi i mužjaci. U najvećem broju odrasli C. elegans su 1 mm dugi hermafroditi sa reproduktivnim ciklusom od 2,5-4 dana na sobnoj temperaturi i srednjim životnim vekom od približno 18-20 dana na 20°C. Nakon izleganja, C. elegans može ili da se razvije direktno kroz četiri stadijuma larve (L1-L4) ili da nastavi sa stadijumom „dauer“ larve, umesto stadijuma L3. Faza „dauer“ larve je razvojno zaustavljena faza disperzije koja se koristi za preživljavanje u nepovoljnim uslovima. Kada se nepovoljni uslovi povuku, C. elegans se može oporaviti i razviti u stadijum larve L4. C. elegans se intenzivno koristi u ispitivanju apoptoze, transdukcije, starenja, zelenog fluorescentnog proteina i njegove uloge u ekspresiji gena, ispitivanju jonskih kanala, testiranju modela bolesti, ispitivanju lekova i dr. U našoj laboratoriji C. elegans koristimo u ispitivanjima mehanizma dejstva antinematodnih lekova i mehanizmima razvoja rezistencije nematoda

Lyophilization as a method for pathogens long term preservation

Lyophilization (freeze-drying) is one of the most suitable methods used for a long term preservation of pathogens. The aim of this paper was the application of lyophilization for storage of three significant plant pathogens: Fusarium graminearum, Helminthosporium gramineum, and Pseudomonas syringae pv. gylicinea, respectively. The plant material was collected continuously (during a four year period 2002-2006), depending on a plant development stage, from different localities in Vojvodina. Pathogens were isolated from diseased parts with characteristic symptoms, and placed on nutritive media specific for a certain pathogen, using standard phytopathological methods. Lyophilization was carried out in marked and coded ampoules by freezing and drying of pathogen suspension and nutritive medium. Revitalization of lyophilized isolates was done after four days. High percentage of revitalization was characteristic for all studied isolates, and it ranged from 85-92%, confirming that lyophilized pathogens would be capable of keeping viability for a long time in the collection. Besides above mentioned pathogens, there were 200 isolates in the collection, originating mostly from field and vegetable crops. Each isolate that was put into the Collection, was followed by all the necessary data such as: name of the pathogen, number of isolates, locality, host plant year of isolation, name of the researcher and other relevant data

TRPV1: A Promising drug target for the treatment of various conditions

Nobelova nagrada za fiziologiju i medicinu za 2021. dodeljena je naučnicima koji su identifikovali receptore za temperaturu i dodir. Da bi sproveli ovo ispitivanje, istraživači su koristili kapsaicin, jedinjenje koje čili paprici daje toplotu, kako bi otkrili receptore koji omogućavaju ljudima da osete “gorući” ukus čilija. Pokazano je da se transient receptor potential vanilloid 1 (TRPV1), koji predstavlja jonski kanal prisutan na senzornim neuronima, otvara u prisustvu kapsaicina ili toplote, propuštajući naelektrisane jone kalcijuma u ćeliju. Takav priliv kalcijuma pokreće električne signale koji se šalju u mozak da upozore na toplotu. TRPV1 se nalazi u somatosenzornom sistemu i služi kao multimodalni senzor različitih štetnih stimulusa. Brojne farmakološke i genetičke studije su potvrdile TRPV1 kao terapeutsku metu u nekoliko pretkliničkih modela hroničnog bola, uključujući maligni, neuropatski, postoperativni i mišićno-skeletni bol. Pored toga, ekspresija TRPV1 se takođe primećuje na ne-neuronskim lokalizacijama, kao što su epitel bešike i pluća, ćelije kohlee u uhu. Stoga, lekovi koji mogu da modulišu aktivnost kanala TRPV1 mogu biti korisni za lečenje različitih stanja u rasponu od hroničnog bola do gubitka sluha. Iako je utvrđeno da antagonisti TRPV1 mogu predstavljati važan dodatak terapiji bola, njihova klinička upotreba je i dalje ograničena značajnim neželjenim efektima, kao što je hipertermija. U ovom radu ćemo opisati najvažnije uloge TRPV1 u fiziološkim i patofiziološkim procesima i predstaviti najperspektivnije lekove koji deluju preko TRPV1.The 2021. Nobel Prize in Physiology or Medicine was awarded to scientists who have identified receptors for temperature and touch. In order to conduct this examination, researchers used capsaicin, the compound that gives chili peppers their heat, to discover receptor proteins that allow people to feel chili’s burn. It is shown that transient receptor potential vanilloid 1 (TRPV1), which represents an ion channel present on sensory neurons, opens when it encounters capsaicin or heat, allowing charged calcium ions into the cell. That flood of calcium triggers electrical signals that are sent to the brain to warn of heat. TRPV1 is found in the somatosensory system and serves as a multimodal sensor of different noxious stimuli. Numerous pharmacological and genetic studies have validated TRPV1 as a therapeutic target in several preclinical models of chronic pain, including cancer, neuropathic, postoperative and musculoskeletal pain. Additionally, expression of TRPV1 is also observed in non-neuronal sites such as the epithelium of bladder and lungs, cells of the cochlea. Therefore, drugs which could modulate TRPV1 channel activity could be useful for the treatment of conditions ranging from chronic pain to hearing loss. While antagonists of TRPV1 were found to be a valuable addition to therapy of pain, their clinical use has still been limited by significant side effects, such as hyperthermia. In this review, we will describe the most important roles of TRPV1 in physiological and pathophysiological processes and present the most promising TRPV1-targeted drugs

Occurence of Fusarium equiseti Corda (Saccardo) as causal agent of seed rot of Hyssopus officinalis L.

Hyssop (Hyssopus officinalis L.) is a perennial polymorphous plant species with essential oil mainly accumulated in the flowers and leaves. It is grown in Serbia for the needs of pharmaceutical companies and tea production, because of its quality and chemical composition. During a routine quality control of hyssop seeds collected from Rumenka (Vojvodina Province), in 2018, fungal infection followed by seed rot was noticed on an average of 22%. Infected seeds were covered with white mycelium followed with violet pigmentation occurring under the seeds. The presence of Fusarium spp. was confirmed with microscopic observation. Isolation was done aseptically by arranging infected seeds onto surface of potato dextrose agar (PDA), and incubated at 25 °C with a 12-h photoperiod (Mathur and Kongsdall, 2003). After seven days, 12 Fusarium spp. isolates were designated as JBL 4003/1 - 4003/12. Pathogenicity test was performed in vitro using a modified agar slant method in the test tube with PDA amended. After 10 days, fungal mycelia of tested isolates caused seed rot and seedling decay, like naturally infected hyssop seeds. All isolates were re-isolated and sub-cultured on Potato Dextrose Agar (PDA) and Carnation Leaf Agar (CLA) using a hyphal tip transfer technique, fulfilling Koch's postulates. Isolate JBL 4003/1 was distinguished based on pathogenicity and cultural characteristics. It caused seed rot after four days, on PDA colony was fast growing reaching 6-8 cm in diam. in five days, forming abundant, whitish to peach aerial mycelium followed with beige to light brown pigmentation in agar

PVDF-HFP/NKBT composite dielectrics: Perovskite particles induce the appearance of an additional dielectric relaxation process in ferroelectric polymer matrix

Na0.25K0.25Bi0.5TiO3 (NKBT) perovskite particles are synthesized by solid-state method and used as a filler for polyvinylidene fluoride-co-hexafluoropropylene (PVDF-HFP) co-polymer. X-ray diffraction analysis of NKBT powders shows that the particles have a rhombohedral perovskite crystal structure (R3c symmetry). Raman spectroscopy reveals that the co-polymer crystallizes predominantly into the mixture of polar beta- and gamma-crystals, while there is also a contribution of the non-polar a-crystal phase. The introduction of the NKBT into the PVDF-HFP results with an increase in effective dielectric permittivity and this effect depends on the inorganic content in the composite. The most interesting result of the present study is that the introduction of NKBT particles induces the appearance of an additional transition peak in the dielectric spectra of the co-polymer matrix. At the fixed frequency of similar to 2 kHz, the observed process appears at similar to 10 degrees C (about 45 degrees above the glass transition temperature) and its magnitude strongly depends on the amount of the NKBT in the composite. Dielectric spectroscopy measurements of the composites are carried out in the wide range of frequencies (from 0.1 Hz to 1 MHz) and temperatures (from -100 to 100 degrees C). They reveal that the novel process can be clearly distinguished in the frequency range between 160 Hz and similar to 50 kHz

Nove NS sorte povrtarskih biljnih vrsta

Povrće je važan deo ljudske ishrane jer ono prvenstveno sadrži materije korisne za ljudsko zdravlje kao što su: vitamini, minerali, biljna vlakna, antioksidansi, organske kiseline i dr. U svetu je poznato oko hiljadu vrsta povrća, od kojih se gaji oko 150, a najširu upotrebu ima 30-50 vrsta, koliko je zastupljeno u ishrani u Srbiji (Gvozdanovic-Varga i sar., 2016). U Institutu za ratarstvo i povrtarstvo Novi Sad do sada je stvoreno više od 50 sorti povrća priznatih u zemlji i delom u inostranstvu. Sortiment povrća Odeljenja za povrtarske i alternativne biljne vrste čine nove sorte povrća, kao i odomaćene sorte, te se proizvodi seme 27 povrtarskih vrsta i 10 vrsta cveća. Pored rada na novim sortama povrća koje se odlikuju visokim prinosom, vodilo se računa da sorte svojim izgledom i ukusom zadovoljavaju zahteve probirljivog tržišta i da su prilagođene našim agroklimatskim uslovima (Gvozdenovic i sar., 2008). Zadatak ovog rada je da se prikažu karakteristike novopriznatih sorti povrća, koje su dobijene kao rezultat višegodišnjeg rada, u cilju zadovoljenja potreba proizvođača i potrošača za novim, atraktivnim i kvalitetnim sortama povrtarskih vrsta

Economic aspects of hospital treated pneumococcal pneumonia and the results of Pneumo 23 vaccine use in Serbia

INTRODUCTION In Serbia, there is a significant number of persons suffering of pneumococcal pneumonia. Persons aged 65 years or older, immunocompromised patients, patients with co-morbidities, such as chronic obstructive lung disease and congestive heart failure, are at the highest risk for developing pneumococcal pneumonia. Most of the patients are treated empirically, although it is often overlooked that Streptococcus pneumoniae can be resistant to the used antibiotics. The treatment costs of such inpatients and outpatients are very high. In Serbia, immunization of persons at risk to develop the diseases caused by Streptococcus pneumoniae is carried out using pneumococcus polysaccharide vaccine according to clinical indications. The exact number of immunized persons and the total number of registered patients are still unknown, but it is certain of being unjustifiably low. OBJECTIVE The goal of the study was to investigate, during a one-year period, the number and basic characteristics of persons hospitably treated for pneumonia, the type of cause of the infection, applied antibiotic medications, duration and costs of hospital treatment at the Institute for Lung Diseases and Tuberculosis of the Clinical Centre of Serbia in Belgrade. METHOD We retrospectively analyzed the medical records of patients with pneumonia treated at the Institute for Lung Diseases and Tuberculosis of the Clinical Centre of Serbia in Belgrade during 2006. RESULTS During the observed one-year period, 290 patients underwent hospital treatment, of whom the cause of the infection was confirmed in 116 (40%). The average duration of hospitalization was 12 days, with treatment cost of 32,031.74 RSD (402.42 EUR) per patient. The treatment cost per patient including general and intensive care was 18,290.01 RSD (229.78 EUR). The distribution cost of Pneumo 23 vaccine in Serbia, without purchase tax, was 746.90 RSD (9.38 EUR). CONCLUSION Pneumococcal pneumonia is a significant medical and economic problem for the healthcare system of Serbia. The use of antipneumococcal vaccination can be useful in decreasing the overall treatment costs related to pneumococcal infection

Detection of viruses transmitted by tomato seeds using serological methods

Tomato (Lycopersicon esculentum) is a plant species parasitized by a great number of microorganisms. Viruses play a significant role regarding the way and speed of transmission, as well as systemic nature of the infection. The aim of this investigation was to determine the presence of two the most significant viruses on tomato Tomato mosaic virus {ToMV) and Tomato spotted wilt virus (TSWV). ToMV is widespread on all tomato growing areas. ToMV is a very stable and can persist in dry contaminated soil, in infected tomato debris, on or in the seed coat. The Tomato spotted wilt virus also has a wide host range and can affect a number of ornamental plants as well as tomato. Early symptoms of spotted wilt on tomato are difficult to diagnose which is very important for nursery plants production. ELISA test is one of the rapid diagnostics techniques for detection of plant viruses. Samples of tomato plants with symptoms were collected from different localities in Vojvodina. Plant samples were tested for the presence of viruses by DAS ELISA test, using specific antibodies purchased from LOEWE Biochemica GmbH, including positive and negative control for both viruses individually

Liofilizacija kao metod dugoročnog čuvanja patogena

Lyophilization (freeze-drying) is one of the most suitable methods used for a long term preservation of pathogens. The aim of this paper was the application of lyophilization for storage of three significant plant pathogens: Fusarium graminearum, Helminthosporium gramineum, and Pseudomonas syringae pv. gylicinea, respectively. The plant material was collected continuously (during a four year period 2002-2006), depending on a plant development stage, from different localities in Vojvodina. Pathogens were isolated from diseased parts with characteristic symptoms, and placed on nutritive media specific for a certain pathogen, using standard phytopathological methods. Lyophilization was carried out in marked and coded ampoules by freezing and drying of pathogen suspension and nutritive medium. Revitalization of lyophilized isolates was done after four days. High percentage of revitalization was characteristic for all studied isolates, and it ranged from 85-92%, confirming that lyophilized pathogens would be capable of keeping viability for a long time in the collection. Besides above-mentioned pathogens, there were 200 isolates in the collection, originating mostly from field and vegetable crops. Each isolate that was put into the Collection, was followed by all the necessary data such as: name of the pathogen, number of isolates, locality, host plant, year of isolation, name of the researcher and other relevant data.Liofilizacija je jedna od najpogodnijih metoda koja se koristi za dugoročno čuvanje patogena. Cilj rada je primena metoda liofilizacije u čuvanju tri značajna biljna patogena: Fusarium graminearum, Helminthosporium gramineum i Pseudomonas syringae pv. gylicinea. Prikupljanje uzoraka biljnog materijala vršeno je u kontinuitetu (tokom četiri godine, 2002-2006) sa različitih lokaliteta na području Vojvodine, u zavisnosti od razvojne faze biljaka. Izolacija patogena vršena je iz obolelih biljnih delova sa karakterističnim simptomima na hranljive podloge specifične za određenog patogena korišćenjem standardnih fitopatoloških metoda