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Biomedical subjects

M Jovanovic

Publications and source records attributed to M Jovanovic.

At least 19 recordsLinked to original sources

miRNAs and apoptosis: RNAs to die for.

MicroRNAs (miRNAs) are small non-coding RNAs of about 18-24 nucleotides in length that negatively regulate gene expression. Discovered only recently, it has become clear that they are involved in many biological processes such as developmental timing, differentiation and cell death. Data that connect miRNAs to various kinds of diseases, particularly cancer, are accumulating. miRNAs can influence cancer development in many ways, including the regulation of cell proliferation, cell transformation, and cell death. In this review, we focus on miRNAs that have been shown to play a role in the regulation of apoptosis. We first describe in detail how Drosophila has been utilized as a model organism to connect several miRNAs with the cell death machinery. We discuss the genetic approaches that led to the identification of those miRNAs and subsequent work that helped to establish their function. In the second part of the review article, we focus on the involvement of miRNAs in apoptosis regulation in mammals. Intriguingly, many of the miRNAs that regulate apoptosis have been shown to affect cancer development. In the end, we discuss a virally encoded miRNA that influences the cell death response in the mammalian host cell. In summary, the data gathered over the recent years clearly show the potential and important role of miRNAs to regulate apoptosis at various levels and in several organisms.

Animals↗

Regulation of TRAIL-induced apoptosis by XIAP in pancreatic carcinoma cells.

Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a promising candidate for cancer therapy because of its relative tumor selectivity. However, many cancers including pancreatic cancer remain resistant towards TRAIL. To develop TRAIL for cancer therapy of pancreatic carcinoma, it will therefore be pivotal to elucidate the molecular mechanisms of TRAIL resistance. Here, we identify X-linked inhibitor of apoptosis (XIAP) as a regulator of TRAIL sensitivity in pancreatic carcinoma cells. Full activation of effector caspases, loss of mitochondrial membrane potential and cytochrome c release following TRAIL treatment were markedly impaired in pancreatic carcinoma cell lines, which poorly responded to TRAIL (PaTuII, PancTu1, ASPC1, DanG), compared to TRAIL-sensitive Colo357 pancreatic carcinoma cells. Stable downregulation of XIAP by RNA interference significantly reduced survival and enhanced TRAIL-induced apoptosis in pancreatic carcinoma cells. Also, downregulation of XIAP significantly increased CD95-induced cell death. Importantly, knockdown of XIAP strongly inhibited clonogenicity of pancreatic cancer cells treated with TRAIL indicating that XIAP promotes clonogenic survival of pancreatic carcinoma cells. Thus, our findings for the first time indicate that targeting XIAP represents a promising strategy to enhance the antitumor activity of TRAIL in pancreatic cancer, which has important clinical implications.

Apoptosis↗

Standardization of transrectal ultrasound-guided biopsy.

Ultrasound-guided biopsy is more sensitive to biopsy performed under the digital control, because 29% of prostatic cancers are not palpable. On the other hand, at least 30% of cancers are isoechogenic, so they cannot be viewed by transrectal ultrasound examination. It means that target biopsy is not sufficient for diagnosis of localized prostatic cancer, i.e., randomized samples are needed as well. More than ten years ago, the technique of sampling the six specimens became a standard procedure to which previously harvested target specimens from suspected growths were added. Today, the expansion of biopsy protocol is recommended, by obtaining the additional specimens from peripheral lateral area, four plus two samples if the prostate has volume over 50 ml. Larger number of biopsies requires anesthesia. In order to reduce complication, the cleaning of rectal ampulla and prophylactic use of quinolone are suggested.

Biopsy, Needle↗

Correlation of high-molecular cytokeratin in tissue of prostatic cancer with Gleason score and PSA.

The absence of basal cell layer of prostatic acini containing high-molecular cytokeratin, which is immunohistochemically detected by monoclonal antibody 34betaE12, is an essential diagnostic characteristic of prostatic cancer. The absence of immunohistochemical reaction in 3 or more pseudoglandular structures of prostatic tissue indicates malignant process. The percentage of immunohistochemically completely negative glandular structures was determined by semiquantitative measurement in tissue specimens obtained by TRUS biopsy of the prostate, and it was correlated with serum PSA concentration and Gleason score. The increase of percentage of glandular prostatic formations completely negative to high-molecular cytokeratin detected by 34betaE12 led to simultaneous rise of mean value of Gleason prostatic cancer score (p < 0.001) as well as the average serum PSA concentration in subjects (p < 0.05).

Adenocarcinoma↗

Glycoprotein identification and localization of O-glycosylation sites by mass spectrometric analysis of deglycosylated/alkylaminylated peptide fragments.

In-gel digestion of densely O-glycosylated proteins, an essential step in proteome analysis, is often hampered by steric hindrance of the proteases. To overcome this technical problem a simple and convenient method has been developed, which combines several advantages: (1) Approximately 70% of the oligosaccharides are cleaved without significant protein hydrolysis at the optimal reaction conditions of 70% ethylamine, and quantitative cleavage is achieved with 40% methylamine, at 50 degrees C. (2) To the unsaturated derivatives of Ser and Thr the alkylamine is added as a label of previous O-glycosylation sites. (3) The alkylaminylated protein is effectively cleaved by proteolysis. (4) The modified peptides are identified by MALDI mass spectrometry under consideration of incremental mass increases. (5) The alkylamine label is stable under MALDI post-source-decay analysis as well as in collision-induced dissociation experiments allowing sequencing and peptide localization of O-glycosylation sites. Applicability of the method is evaluated with a series of synthetic glycopeptides, the densely O-glycosylated human glycophorin A, and with the mucin MUC1 from human milk fat globule membranes.

Alkylation↗

Tuberculosis trends in Central Serbia in the period 1956-1996.

OBJECTIVE: To estimate tuberculosis incidence and mortality trends in Central Serbia (excluding Kosovo and Vojvodina provinces) in the period 1956-1996. DESIGN: The incidence and mortality data of tuberculosis in Central Serbia in the period 1956-1996 were analysed based on the annual reports of the Institute for Lung Diseases and Tuberculosis in Belgrade and the official data of the Republic Health Institute. RESULTS: During the period under observation, tuberculosis incidence decreased from 324.0 to 34.8/100 000 population, fitting the exponential model (y = 389.066e-(0.0689); F = 847.60; P = 0.000). Mortality rates decreased from 76.0 in 1956 to 0.9/100000 in 1982. Over the whole period (1956-1996) the decrease in mortality rates fitted the exponential model (y = 66.83e-(0.0922); F = 150.95; P = 0.000). The increase in mortality rates in the period 1982-1996 fitted cubic model (y = 7.647 - 2.674x + 0.359x(2)-0.013x(3); F = 12.17; P = 0.001). CONCLUSION: Decreasing trends in tuberculosis incidence are the result of good control programmes. Changes in mortality trends are related to migration from war zones and poor economic conditions which hinder the detection and treatment of tuberculosis.

Female↗

Occupational risk of tuberculosis among health care workers at the Institute for Pulmonary Diseases of Serbia.

SETTING: The Institute for Pulmonary Diseases of Serbia, which specialises in diagnosis and treatment of lung diseases in a region with approximately 550-600 TB admissions per year. OBJECTIVE: To assess the occupational risk of tuberculosis (TB) among health care workers (HCWs) employed at this institution. DESIGN: Retrospective analysis of morbidity of TB among HCWs over a 12-year period (1986-1997). The incidence of TB among HCWs was estimated by the frequency of bacteriologically or histologically confirmed active disease. All HCWs at our institution underwent routine pre-employment screening, consisting of verification of BCG vaccination, PPD tuberculin reactivity, chest X-ray and laboratory evaluation. RESULTS: Of an average 267 employed HCWs, pulmonary TB occurred in nine (six nurses and three laboratory technicians). Cumulative incidence for HCWs was 3451/100,000, compared to 454/100,000 in the general population, for an incidence rate ratio of 7.6. CONCLUSION: The risk of TB among HCWs employed at the Institute for Pulmonary Diseases of Serbia in Belgrade is 7.6 times higher than that observed in the general population, suggesting occupational acquisition of TB.

Adult↗

tRNA synthetase mutants of Escherichia coli K-12 are resistant to the gyrase inhibitor novobiocin.

In previous studies we demonstrated that mutations in the genes cysB, cysE, and cls (nov) affect resistance of Escherichia coli to novobiocin (J. Rakonjac, M. Milic, and D. J. Savic, Mol. Gen. Genet. 228:307-311, 1991; R. Ivanisevic, M. Milic, D. Ajdic, J. Rakonjac, and D. J. Savic, J. Bacteriol. 177:1766-1771, 1995). In this work we expand this list with mutations in rpoN (the gene for RNA polymerase subunit sigma54) and the tRNA synthetase genes alaS, argS, ileS, and leuS. Similarly to resistance to the penicillin antibiotic mecillinam, resistance to novobiocin of tRNA synthetase mutants appears to depend upon the RelA-mediated stringent response. However, at this point the overlapping pathways of mecillinam and novobiocin resistance diverge. Under conditions of stringent response induction, either by the presence of tRNA synthetase mutations or by constitutive production of RelA protein, inactivation of the cls gene diminishes resistance to novobiocin but not to mecillinam.

Alanine-tRNA Ligase↗

Percutaneous diagnostic stabilization for low back pain. Correlation with results after fusion operations.

Diagnostic external fixation was performed in 101 patients with disabling low back pain. In 47 patients, pain was relieved by stabilization but returned after destabilization. These patients were considered good candidates for a fusion operation. Results after fusion are available for 34 patients: 14 (41%) patients had a good, 12 (35%) had a fair, and 8 (14%) had a bad result. In two patients, pain was relieved by stabilization and did not return after fixator removal; no fusion operation was performed. Fifty-two patients did not respond positively to external fixation. Nine were operated on despite negative results with fixation. Of these, seven patients had a bad result, one a good result (however, this patient had spinal stenosis and the indication for external fixation was wrong), and, in one, the follow-up time is too short. Positive results with external skeletal fixation may predict a successful fusion operation with reasonable accuracy. If stabilization does not relieve the patient's pain, spinal fusion is unlikely to be of any benefit.

Adult↗

Tissue response and in vivo degradation of selected polyhydroxyacids: polylactides (PLA), poly(3-hydroxybutyrate) (PHB), and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB/VA).

The tissue response and in vivo molecular stability of injection-molded polyhydroxyacids--polylactides (PLA), poly(3-hydroxybutyrate) (PHB), and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB/VA, 5-22% VA content)--were studied. Polymers were implanted subcutaneously in mice and extirpated at 1, 3, and 6 months in order to study tissue response and polymer degradation. All polymers were well tolerated by the tissue. No acute inflammation, abscess formation, or tissue necrosis was observed in tissues adjacent to the implanted materials. Furthermore, no tissue reactivity or cellular mobilization was evident remote from the implant site. Mononuclear macrophages, proliferating fibroblasts, and mature vascularized fibrous capsules were typical of the tissue response. Degradation of the polymers was accompanied by an increase in collagen deposition. For the polylactide series, the inflammatory response after 1 month of implantation was less for materials containing the D-unit in the polymer chain, whereas in the case of the polyhydroxybutyrate/valerates, the number of inflammatory cells increased with increasing content of the valerate unit in the polymer chain. Between 1-3 months, there was slightly more tissue response to the PHB and PHB/VA polymers than to PLA. This response is attributed to the presence of leachable impurities and a low molecular weight soluble component in the polyhydroxybutyrate/valerates. At 6 months, the extent of tissue reaction was similar for both types of polymers. All polylactides degraded significantly (56-99%) by 6 months. For a poly(L-lactide) series, degradation rate in vivo decreased with increasing initial molecular weight of the injection-molded polymer. Several samples showed pronounced bimodal molecular weight distributions (MWD), which may be due to differences in degradation rate, resulting from variability in distribution of crystalline and amorphous regions within the samples. This may also be the result of two different mechanisms, i.e., nonenzymatic and enzymatic, which are involved in the degradation process, the latter being more extensive at the later stage of partially hydrolyzed polymer. The PHB and PHB/VA polymers degraded less (15-43%) than the polylactides following 6 months of implantation. Generally, the polymer with higher valerate content (19%, 22%) degraded most. The decrease in molecular weight was accompanied by a narrowing of the MWD for PHB and copolymers; there was no evidence of a bimodal MWD, possibly indicating that the critical molecular weight that would permit enzyme/polymer interaction had not been reached. Weight loss during implantation ranged from 0-50% for the polylactides, whereas for the PHB polymers weight loss ranged from 0-1.6%.

Absorption↗

Orientation and moment arms of some trunk muscles.

The orientations and moment arms of nine trunk muscles: latissimus dorsi, iliocostalis, longissimus dorsi, obliquii, rectus and transversalis abdominis, quadratus lumborum, and iliopsoas were obtained on embalmed specimens. Each muscle was considered as one or several, linear or curvilinear, lines of action. Origins, insertions, and points along the length of main bundles were digitized for each muscle using a three-dimensional digitizer. The results obtained from seven male specimens are presented and will permit more refined biomechanical models.

Abdominal Muscles↗

3C syndrome: third occurrence of cranio-cerebello-cardiac dysplasia (Ritscher-Schinzel syndrome).

We report a child with an unusual pattern of malformations: severe delay in bone maturation, wide fontanelles and facial dysmorphism (evoking cleidocranial dysplasia), relative macroencephaly with cerebellar vermis hypoplasia, hypertelorism, skeletal abnormalities (1st ribs aplasia, multifocal sternal ossification centers, thin bones), septal defect, muscular waste, hypotonia and developmental delay. Most of these features have been reported previously by Ritscher, Schinzel et al. in two sibs, who suffered more severe cerebellar malformations (Dandy-Walker cyst or vermis aplasia). We propose 3C syndrome as an easy acronym for this Cranio-Cerebello-Cardiac dysplasia.

Abnormalities, Multiple↗

A three-dimensional digitization method to measure trunk muscle lines of action.

A method is presented to obtain lines of action of nine major trunk muscles acting on the lumbar spine. The muscles are modeled as one or several linear or curvilinear lines of action obtained by digitizing the origins and insertions of main bundles of muscles. For curvilinear lines of action, points were also digitized along the length of the bundles, and a curve-fitting method was used. Lines of action are displayed graphically for one embalmed cadaver. Tabulated data for unit force vectors and unit moment vectors about the L3-L4 and L4-L5 discs are also presented.

Adult↗