Tehnologija uzgoja dagnji (mytilus galloprovincialis) i kamenica (ostrea edulis) u Bokokotorskom zalivu

Razvoj marikulture u Crnoj Gori je jedno od strateški važnih pitanja u programima proizvodnje hrane za domaće tržište i za izvoz. Program podrazumijeva proizvodnju, odnosno uzgoj nekoliko vrsta školjaka i riba, dok je u planu i uzgoj rakova. Marikultura je sve značajnija privredna grana u svijetu jer nadoknađuje smanjene potencijale hrane iz prirodnih izvora. I pored naglašenog značaja marikulture, izuzetnih bioloških i ekoloških karakteristika priobalnog mora Crne Gore, naučnog-stručnog znanja, moramo konstatotovati da je uzgoj morskih organizama na navedenom području tek u inicijalnom začetku, čak i kada se radi o uzgoju školjaka, čija je sadašnja proizvodnja oko 200 t. godišnje, što ostvaruje 16 uzgajivača u Bokokotorskom zalivu- uglavnom u Kotorskom i Tivatskom dijelu. Dakle, cijela marikultura u Crnoj Gori koncentrisana je na područje Bokokotorskog zaliva, dok je otvoreno more, u tom smislu, ostalo potpuno neiskorišćeno. U davna vremena način uzgoja školjaka bio je vrlo jednostavan. Grane hrasta, trešnje, masline i ostalog raspoloživog materijala su se sakupljale i bacale u more. Na tako bačene grane hvatala se mlađ školjaka (kamenica i dagnji), a nakon tri godine i nekoliko faza uzgoja proces je bio kompletan, školjke su se vadile iz mora, te otpremale na tržište. Cilj ovog rada je da damo svojevrsni pregled, počevši od prvih istraživanja mogućnosti uzgoja dagnji i kamenica u Bokokotorskom zalivu do današnjeg stanja i pravaca u kojima bi marikultura trebala da se razvoja u Crnoj Gori

Charting calcium-regulated apoptosis pathways using chemical biology: role of calmodulin kinase II

<p>Abstract</p> <p>Background</p> <p>Intracellular free calcium ([Ca²⁺]_i) is a key element in apoptotic signaling and a number of calcium-dependent apoptosis pathways have been described. We here used a chemical biology strategy to elucidate the relative importance of such different pathways.</p> <p>Results</p> <p>A set of 40 agents ("bioprobes") that induce apoptosis was first identified by screening of a chemical library. Using p53, AP-1, NFAT and NF-κB reporter cell lines, these bioprobes were verified to induce different patterns of signaling. Experiments using the calcium chelator BAPTA-AM showed that Ca²⁺was involved in induction of apoptosis by the majority of the bioprobes and that Ca²⁺was in general required several hours into the apoptosis process. Further studies showed that the calmodulin pathway was an important mediator of the apoptotic response. Inhibition of calmodulin kinase II (CaMKII) resulted in more effective inhibition of apoptosis compared to inhibition of calpain, calcineurin/PP2B or DAP kinase. We used one of the bioprobes, the plant alkaloid helenalin, to study the role of CaMKII in apoptosis. Helenalin induced CaMKII, ASK1 and Jun-N-terminal kinase (JNK) activity, and inhibition of these kinases inhibited apoptosis.</p> <p>Conclusion</p> <p>Our study shows that calcium signaling is generally not an early event during the apoptosis process and suggests that a CaMKII/ASK1 signaling mechanism is important for sustained JNK activation and apoptosis by some types of stimuli.</p

Memory and relatedness of transcriptional activity in mammalian cell lineages

Phenotypically identical mammalian cells often display considerable variability in transcript levels of individual genes. How transcriptional activity propagates in cell lineages, and how this varies across genes is poorly understood. Here we combine live-cell imaging of short-lived transcriptional reporters in mouse embryonic stem cells with mathematical modelling to quantify the propagation of transcriptional activity over time and across cell generations in phenotypically homogenous cells. In sister cells we find mean transcriptional activity to be strongly correlated and transcriptional dynamics tend to be synchronous; both features control how quickly transcriptional levels in sister cells diverge in a gene-specific manner. Moreover, mean transcriptional activity is transmitted from mother to daughter cells, leading to multi-generational transcriptional memory and causing inter-family heterogeneity in gene expression

Proučavanje funkcionalnih profila mikrobnih zajednica zagađenih zemljišta

In this review paper are considerate the different ways of soil microorganism’s reaction in a variety of forms of technogenic pollution by methods multi-substrate testing and initiates the microbial community, as well as the modification of variability that is characterized by specific adaptive reactions (homeostasis zone, stress zone, zone of resistance and zone repression) of microbial system lands.U ovom pregednom radu se razmatraju načini sagledavanja reakcije zemljišnih mikroorganizama na različite oblike tehnogenog zagađenja pomoću metoda multisupstratnog testiranja i inicirane mikrobne zajednice, kao i modifikacione promenljivosti koja se odlikuje određenom adaptivnom reakciijom (zona homeostaze, zona stresa, zona rezistencije i zona represije) mikrobnog sistema zemljišta

PERCEPTIONS ON SUPPORT, CHALLENGES AND NEEDS AMONG PARENTS OF CHILDREN WITH AUTISM: THE SERBIAN EXPERIENCE

Background: Autism spectrum disorders (ASD) significantly impact lives of affected individuals and their families. They confront daunting challenges and multiple demands in their daily life, when compared to parents of children with other disabilities or parents of typically developing children. Subjects and methods: Participants completed The Caregiver Needs Survey, the survey intended for parents or primary caregivers of children with a diagnosis of ASD. During the study, 231 parents were interviewed; 167 mothers and 64 fathers. Parents were recruited from the patient database comprised of families from the two largest cities in Serbia. All of them were contacted before the study, either via phone or at the child’s regular check-in visit. Results: Over 90 percent of the parents reported that additional support at schools, home, and improved relationships with service providers are necessary and important. The most important challenges related to care were child’s communication difficulties, social interaction difficulties, and problems with daily living skills. The significant predictors of lower overall satisfaction were parent’s higher education, having a first concern related to problems of the child’s interaction with others or playing alone, and parent frustration with accessing services in the past 12 months. Greater overall satisfaction, on the other hand, was related to having an in-school tutor training or assistance in managing child’s needs or implementing treatments, and having primary care doctor or pediatrician as a source of information on autism. Conclusions: Future efforts to develop ASD-related policies and services should also take the following into consideration: the low level of awareness among caregivers and health care providers about the early signs of autism; disparities in access to services; educational problems and significant levels of dissatisfaction with the overall care and stigma

PERCEPTIONS ON SUPPORT, CHALLENGES AND NEEDS AMONG PARENTS OF CHILDREN WITH AUTISM: THE SERBIAN EXPERIENCE

Background: Autism spectrum disorders (ASD) significantly impact lives of affected individuals and their families. They confront daunting challenges and multiple demands in their daily life, when compared to parents of children with other disabilities or parents of typically developing children. Subjects and methods: Participants completed The Caregiver Needs Survey, the survey intended for parents or primary caregivers of children with a diagnosis of ASD. During the study, 231 parents were interviewed; 167 mothers and 64 fathers. Parents were recruited from the patient database comprised of families from the two largest cities in Serbia. All of them were contacted before the study, either via phone or at the child’s regular check-in visit. Results: Over 90 percent of the parents reported that additional support at schools, home, and improved relationships with service providers are necessary and important. The most important challenges related to care were child’s communication difficulties, social interaction difficulties, and problems with daily living skills. The significant predictors of lower overall satisfaction were parent’s higher education, having a first concern related to problems of the child’s interaction with others or playing alone, and parent frustration with accessing services in the past 12 months. Greater overall satisfaction, on the other hand, was related to having an in-school tutor training or assistance in managing child’s needs or implementing treatments, and having primary care doctor or pediatrician as a source of information on autism. Conclusions: Future efforts to develop ASD-related policies and services should also take the following into consideration: the low level of awareness among caregivers and health care providers about the early signs of autism; disparities in access to services; educational problems and significant levels of dissatisfaction with the overall care and stigma

Inficiranje leguminoznih biljaka sa kvržičnim bakterijama

In this review paper we are discussing the question of the expediency of biological symbiosis between leguminous plants and rhisobium. In addition, we give the basic properties of symbionts interactions that characterize this complex system, as well as the dependence of this type of partnership of environmental conditions. In particular, they are viewed special stages of infecting plants with rhisobium. Finally, it is concluded that the effectiveness of the symbiosis between legumes and rhisobium depends not only of biological but also from abiotical factors; that this symbiosis is characterized by a relatively rapid return reactions to external influences, especially in the first stages of establishing symbiosis; that formed system is quite stable and can function effectively even in unfavorable environmental conditions.U ovom radu daje se osvrt na pitanje biološke svrsishodnosti simbioze između leguminoznih biljaka i rizobijuma. Osim toga, navode se osnovna svojstva interakcije simbionata koja karakterišu ovaj složeni sistem, kao i zavisnost ovog tipa partnerstva od ekoloških uslova sredine. Posebno se sagledavaju etape inficiranja biljke sa rizobijumima. Na kraju se zaključuje da efektivnost simbioze između leguminoza i rizobijuma ne zavisi samo od bioloških, već i od abitičkih faktora; da se ova simbioza odlikuje relativno brzim povratnim reakcijama na spoljašnje uticaje, posebno na prvim etapama uspostavljanja simbioze; da je formirani sistem dosta stabilan i da može efikasno funkcionisati čak i u nepovoljnim ekološkim uslovima

Nickel as a potential disruptor of thyroid function: benchmark modelling of human data

IntroductionNickel (Ni) is one of the well-known toxic metals found in the environment. However, its influence on thyroid function is not explored enough. Hence, the aim of this study was to analyse the potential of Ni to disrupt thyroid function by exploring the relationship between blood Ni concentration and serum hormone levels (TSH, T4, T3, fT4 and fT3), as well as the parameters of thyroid homeostasis (SPINA-GT and SPINA-GD) by using correlation analysis and Benchmark (BMD) concept.MethodsNi concentration was measured by ICP-MS method, while CLIA was used for serum hormone determination. SPINA Thyr software was used to calculate SPINA-GT and SPINA-GD parameters. BMD analysis was performed by PROAST software (70.1). The limitations of this study are the small sample size and the uneven distribution of healthy and unhealthy subjects, limited confounding factors, as well as the age of the subjects that could have influenced the obtained results.Results and discussionThe highest median value for blood Ni concentration was observed for the male population and amounted 8,278 µg/L. Accordingly, the statistically significant correlation was observed only in the male population, for Ni-fT4 and Ni-SPINA-GT pairs. The existence of a dose-response relationship was established between Ni and all the measured parameters of thyroid functions in entire population and in both sexes. However, the narrowest BMD intervals were obtained only in men, for Ni - SPINA-GT pair (1.36-60.9 µg/L) and Ni - fT3 pair (0.397-66.8 µg/L), indicating that even 78.68 and 83.25% of men in our study might be in 10% higher risk of Ni-induced SPINA-GT and fT3 alterations, respectively. Due to the relationship established between Ni and the SPINA-GT parameter, it can be concluded that Ni has an influence on the secretory function of the thyroid gland in men. Although the further research is required, these findings suggest possible role of Ni in thyroid function disturbances

Elucidation of pro-apoptotic signaling induced by cisplatin

A major clinical problem regarding treatment of malignant tumors with anticancer drugs is inherent or acquired resistance to therapy. One factor contributing to drug resistance of cancer cells is failure to undergo apoptosis. The aim of this thesis was to investigate pro-apoptotic signaling induced by the anti-cancer drug cisplatin in order to contribute to future improvement of cisplatin-based cancer therapy. The results demonstrate the ability of cisplatin to induce at least two separate proapoptotic pathways, and to affect several cellular organelles. The ERK and the JNK pathways are the two main mitogen activated protein kinase (MAPK) modules of serial kinase activities stimulated by proliferative signaling as well as by cellular stress. The role of ERK in cisplatin-induced apoptosis was studied in four melanoma cell lines. ERK activation was seen 4 h after cisplatin treatment. Inhibition of the pro survival ERK pathway resulted in different responses, from sensitizing to slightly protective. The cellular response to a certain type of treatment can thus not always be predicted. Cisplatin treatment was found to elicit increased intracellular calcium levels and activation of calpain within 1-3 hours. Involvement of calpain in the apoptotic response was demonstrated using calpain inhibitors which blocked caspase activation and nuclear fragmentation. Moreover, the calpain inhibitor calpeptin was able to block cisplatin-induced cleavage of the proapoptotic Bid. Recombinant Bid was cleaved in vitro by recombinant calpain as well as lysates of cisplatin-treated cells. Apoptosis via the intrinsic (mitochondrial) pathway is known to require activation of the pro-apoptotic Bcl-2 proteins Bak and Bax. We demonstrate the ability of cisplatin to induce activation of Bak, and suggest a kinase fragment of MEKK1 (deltaMEKK1 ) as mediator of Bak activation, since activation was inhibited in cells expressing a kinase-inactive mutant of MEKK1 (dominant negative MEKK1, dnMEKK1). Involvement of deltaMEKK1 in Bak activation was supported by the ability of a constitutively active kinase mutant of MEKK1 (dominant positive MEKK1, dpMEKKI1) to activate Bak in three cell lines out of four, and to induce apoptosis in two of them. In contrast to dnMEKK1, calpeptin did not affect Bak activation but did block Bid cleavage. On the other hand Bid cleavage was not affected by dnMEKK1. Calpeptin and dnMEKK1 were both able to block nuclear fragmentation and caspase activation by approximately half. When used together, they had an additive inhibitory effect on apoptosis. Using enucleated cells (cytoplasts) we demonstrated the ability of cisplatin to induce apoptosis in the absence of nuclear DNA. Cisplatin treatment of cytoplasts resulted in calpainmediated Bid cleavage as well as calcium- and calpain-dependent activation of caspase-3. Moreover, calpeptin- sensitive activation of pro-caspase- 12 was found in cisplatin-treated cells and cytoplasts. Caspase- 12 is localized at the endoplasmic reticulum (ER) and is activated by ER stress. Cisplatininduced ER stress was supported by increased expression of grp78, an ER chaperone protein. In summary, we have characterized two novel pro-apoptotic pathways induced by cisplatin and investigated the time course of their activation. One pathway is activated early and involves calciumdependent activation of calpain and calpain-mediated cleavage of Bid. The other pathway involves deltaMEKK1-mediated activation of Bak shortly before onset of caspase-3. We have also identified ER as a new, non-nuclear target of cisplatin, and suggest ER stress as a potent mediator of cisplatin-induced proapoptotic signaling

Quantitative analysis of gene expression and transcriptional memory in mouse embryonic stem cells

Quantitative studies of gene expression in have shown widespread variability in transcript and protein product abundance at the single-cell level. Such fluctuations in gene expression have been shown to lead to heterogeneous cellular responses and phenotypes, acting in a range of systems, from immunity, cancer to development. To better understand heterogeneity in cellular populations, we developed a novel method for the precise quantification of gene expression in single living cells. Furthermore, we focused on whether and to which extent transcriptional expression profiles are transmitted during cell division. In the first section, we described a new approach for quantitative measurements of gene expression, based on internal tagging of endogenous proteins with a reporter allowing luminescence and fluorescence time-lapse imaging. Using the currently brightest luminescent reporter, fluctuations of protein expression levels could be monitored in single living cells with high sensitivity and temporal resolution over extended time periods. Moreover, our system allowed measurement of single-cell protein degradation rates in the absence of protein synthesis inhibitors, as well as calculation of absolute synthesis rates over the cell cycle. Finally, the internal tag could be excised by inducible expression of Cre recombinase, which enabled estimation of endogenous mRNA half-lives by monitoring the decay of luminescent signal. Second, to monitor transcriptional activity and quantify memory across multiple endogenous genes in mouse embryonic stem cells, we generated cell lines in which a short-lived luciferase reporter allowed sensitive monitoring of transcriptional kinetics by luminescence imaging at high time resolution over long periods of time. By coupling live single-cell monitoring of transcriptional activity of 9 different promoters with mathematical modelling, we found that different levels of expression fluctuations shape transcriptional memory. Long fluctuations (1-10 generations) and short fluctuations (1-3 h), as well as their amplitudes, vary widely between across different genes and together shape transcriptional memory. Surprisingly, we have shown that there was a large level of gene-specific cell-to-cell synchronicity in transcriptional profiles. Overall, we uncovered that noise and timescales of slow and fast fluctuations, as well as their synchronisation over the cell cycle, defined how transcriptional dynamics were correlated within a lineage of related cells. In conclusion, this thesis described an innovative method that allows measurement of absolute protein expression levels, protein turnover, as well as mRNA half-lives for endogenously expressed genes. Our approach opens new avenues in quantitative studies of gene expression in single living cells. Furthermore, we revealed different levels of transcriptional fluctuations that shape gene specific transcriptional memory and its timescales in mouse embryonic stem cells