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A P Burnens,J Wagner,H Lior,J Nicolet,J Frey Several typing systems have been described for Campylobacter jejuni and C. coli, to assess the complex epidemiology of these important enteric pathogens. In the present study two typing methods, slide agglutination according to the Lior scheme, and the demonstration of restriction-fragment length p... ( view more )olymorphisms (RFLP) of flagellar genes, have been used in parallel on a set of 194 strains. This set comprised 118 sero-reference strains of C. jejuni and C. coli of the Lior scheme, as well as 76 clinical isolates. All isolates were serotyped and subjected to PCR for amplification of flagellar genes, and the PCR product was restricted with Alu I. Flagellar genes could be amplified in 152 strains. Among 85 seroreference strains, 74 different RFLP patterns were observed, and among 67 clinical isolates, there were 36 patterns. There was only limited correlation between flagellar RFLP and the Lior serogroup, and the variability of patterns in serogroups HL2 and HL4 were as marked as the variability between serogroups. Flagellar gene RFLP patterns are shown to be stable, highly discriminatory epidemiologic markers. ( view less ) R A Alm,P Guerry,M E Power,H Lior,T J Trust Flagellin mutations originally constructed in Campylobacter coli VC167 (serotype LIO8) by a gene replacement mutagenesis technique (P. Guerry, S. M. Logan, S. Thornton, and T. J. Trust, J. Bacteriol. 172:1853-1860, 1990) were moved from the original host into Campylobacter strains of a number of ot... ( view more )her Lior serogroups by a natural transformation procedure. This is the first report of the use of this transformation method to transfer a mutated locus among Campylobacter strains. Flagellin mutants were constructed in a number of heat-labile LIO serotypes and were serotyped and analyzed by immunoelectron microscopy with LIO typing antisera. In six cases, isogenic nonflagellated mutants were able to be serotyped in the same serogroup as their parent, and immunogold electron microscopy confirmed that antibodies in the typing antisera bound to components on the surface of both parent and mutant cells. However, in only one case, a strain belonging to serogroup LIO4, was a nonflagellated mutant untypeable, and immunogold electron microscopy showed that antibodies bound to the flagella filament of the parent but not to the cell surface. Furthermore, after introduction and expression as a flagellar filament of a LIO8 flagellin gene in this mutant, the strain could not be serotyped. These results indicate that a nonflagellar antigen is often the serodeterminant in the heat-labile Lior serotyping scheme. ( view less ) A D Fraser,B W Brooks,M M Garcia,H Lior Plasmid, protein and restriction endonuclease analysis (REA) profiles and multilocus enzyme electrophoresis were used to effect a molecular discrimination of twenty-seven Campylobacter coli serogroup 20, biotype 1 (Lior) strains. These strains were not outbreak-associated but were isolated from a n... ( view more )umber of different countries and different animal and environmental sources. Each of the techniques was able to discriminate, to various degrees, between the serogroup 20, biotype 1 strains. The choice of a particular technique depends to a large extent on the level of discrimination desired, the previous experiences of the investigator and on the laboratory facilities at hand. REA profiles demonstrated the greatest degree of discrimination between these strains. Plasmid and protein profiles could discriminate reasonably well. Multilocus enzyme electrophoresis (allozyme typing) and protein profiles may prove effective in subgrouping serogroup 20, biotype 1 strains. ( view less ) Lior Yankelson,Yair Feld,Tal Bressler-Stramer,Ilanit Itzhaki,Irit Huber,Amira Gepstein,Doron Aronson,Shimon Marom,Lior Gepstein BACKGROUND: Traditional antiarrhythmic pharmacological therapies are limited by their global cardiac action, low efficacy, and significant proarrhythmic effects. We present a novel approach for the modification of the myocardial electrophysiological substrate using cell grafts genetically engineere... ( view more )d to express specific ionic channels. METHODS AND RESULTS: To test the aforementioned concept, we performed ex vivo, in vivo, and computer simulation studies to determine the ability of fibroblasts transfected to express the voltage-sensitive potassium channel Kv1.3 to modify the local myocardial excitable properties. Coculturing of the transfected fibroblasts with neonatal rat ventricular myocyte cultures resulted in a significant reduction (68%) in the spontaneous beating frequency of the cultures compared with baseline values and cocultures seeded with naive fibroblasts. In vivo grafting of the transfected fibroblasts in the rat ventricular myocardium significantly prolonged the local effective refractory period from an initial value of 84+/-8 ms (cycle length, 200 ms) to 154+/-13 ms (P<0.01). Margatoxin partially reversed this effect (effective refractory period, 117+/-8 ms; P<0.01). In contrast, effective refractory period did not change in nontransplanted sites (86+/-7 ms) and was only mildly increased in the animals injected with wild-type fibroblasts (73+/-5 to 88+/-4 ms; P<0.05). Similar effective refractory period prolongation also was found during slower pacing drives (cycle length, 350 to 500 ms) after transplantation of the potassium channels expressing fibroblasts (Kv1.3 and Kir2.1) in pigs. Computer modeling studies confirmed the in vivo results. CONCLUSIONS: Genetically engineered cell grafts, transfected to express potassium channels, can couple with host cardiomyocytes and alter the local myocardial electrophysiological properties by reducing cardiac automaticity and prolonging refractoriness. ( view less ) Sharon Melamed,Melamed Sharon,Nir Paran,Paran Nir,Lior Katz,Katz Lior,David Ben-Nathan,Ben-Nathan David,Tomer Israely,Israely Tomer,Paula Schneider,Schneider Paula,Reuven Levin,Levin Reuven,Shlomo Lustig,Lustig Shlomo Since smallpox eradication by the WHO during the 1980s, potency of new vaccines is compared to vaccines that were used during the eradication campaign. In this work we characterize the tail scarification technique in mice as a model for scarification in humans. Similar to humans, mice develop "clin... ( view more )ical take" which is dependent on the vaccination dose. Appearance of anti-Vaccinia IgM is followed by IgG antibodies 10 days post scarification and lasting more then 1(1/2) years. Mice with "clinical take" are 100% protected against lethal respiratory challenge (100LD(50)) of Vaccinia WR indicating that the "clinical take" can serve as a correlate of protective immunity. Reducing the vaccination dose and using Cowpox virus as a more virulent strain, enabled us to draw the limit of the vaccine potency in mice. Similar to humans, in revaccinated mice the development of "clinical take" was inversely correlated to the level of pre-existing antibodies. These results indicate that tail scarification of mice can be used as a model for evaluation of smallpox vaccines. High correlation between "clinical take" and protective immunity allows the use of visual inspection to evaluate vaccine potency. ( view less ) Oren Caspi,Irit Huber,Izhak Kehat,Manhal Habib,Gil Arbel,Amira Gepstein,Lior Yankelson,Doron Aronson,Rafael Beyar,Lior Gepstein OBJECTIVES: We evaluated the ability of human embryonic stem cells (hESCs) and their cardiomyocyte derivatives (hESC-CMs) to engraft and improve myocardial performance in the rat chronic infarction model. BACKGROUND: Cell therapy is emerging as a novel therapy for myocardial repair but is hampered ... ( view more )by the lack of sources for human cardiomyocytes. METHODS: Immunosuppressed healthy and infarcted (7 to 10 days after coronary ligation) rat hearts were randomized to injection of undifferentiated hESCs, hESC-CMs, noncardiomyocyte hESC derivatives, or saline. Detailed histological analysis and sequential echocardiography were used to determine the structural and functional consequences of cell grafting. RESULTS: Transplantation of undifferentiated hESCs resulted in the formation of teratoma-like structures. This phenomenon was prevented by grafting of ex vivo pre-differentiated hESC-CMs. The grafted cardiomyocytes survived, proliferated, matured, aligned, and formed gap junctions with host cardiac tissue. Functionally, animals injected with saline or nonmyocyte hESC derivatives demonstrated significant left ventricular (LV) dilatation and functional deterioration, whereas grafting of hESC-CMs attenuated this remodeling process. Hence, post-injury baseline fractional shortening deteriorated by 50% (from 20 +/- 2% to 10 +/- 2%) and by 30% (20 +/- 2% to 14 +/- 2%) in the saline and nonmyocyte groups while improving by 22% (21 +/- 2% to 25 +/- 3%) in the hESC-CM group. Similarly, wall motion score index and LV diastolic dimensions were significantly lower in the hESC-CM animals. CONCLUSIONS: Transplantation of hESC-CMs after extensive myocardial infarction in rats results in the formation of stable cardiomyocyte grafts, attenuation of the remodeling process, and functional benefit. These findings highlight the potential of hESCs for myocardial cell therapy strategies. ( view less ) Irit Huber,Ilanit Itzhaki,Oren Caspi,Gil Arbel,Maty Tzukerman,Amira Gepstein,Manhal Habib,Lior Yankelson,Izhak Kehat,Lior Gepstein Human embryonic stem cells (hESC) are pluripotent lines that can differentiate in vitro into cell derivatives of all three germ layers, including cardiomyocytes. Successful application of these unique cells in the areas of cardiovascular research and regenerative medicine has been hampered by diffi... ( view more )culties in identifying and selecting specific cardiac progenitor cells from the mixed population of differentiating cells. We report the generation of stable transgenic hESC lines, using lentiviral vectors, and single-cell clones that express a reporter gene (eGFP) under the transcriptional control of a cardiac-specific promoter (the human myosin light chain-2V promoter). Our results demonstrate the appearance of eGFP-expressing cells during the differentiation of the hESC as embryoid bodies (EBs) that can be identified and sorted using FACS (purity>95%, viability>85%). The eGFP-expressing cells were stained positively for cardiac-specific proteins (>93%), expressed cardiac-specific genes, displayed cardiac-specific action-potentials, and could form stable myocardial cell grafts following in vivo cell transplantation. The generation of these transgenic hESC lines may be used to identify and study early cardiac precursors for developmental studies, to robustly quantify the extent of cardiomyocyte differentiation, to label the cells for in vivo grafting, and to allow derivation of purified cell populations of cardiomyocytes for future myocardial cell therapy strategies. ( view less ) Lior Koren,Reuven Gurfinkel,Ronen Glezinger,Zvi Howard Perry,Sandra Lev-Ari,Lior Rosenberg Sepsis as a result of bacterial translocation from the gastrointestinal tract (GIT) is a known associate of morbidity and mortality in patients with severe burns. This translocation is influenced by the GIT flora. Oral consumption of Lactobacillus bacteria was previously shown to reduce translocati... ( view more )on. We conducted a retrospective cohort study on a series of 56 patients with burns admitted to Soroka University Medical Center in Beer-Sheva, Israel. Those 56 patients included 28 who were given lactobacillus supplements and 28 who were not. The parameters that were compared between the groups evaluated the level of sepsis and its complications. The parameters of morbidity during hospitalization were significantly higher in the treatment group; however, their mortality was lower. That difference in mortality between the groups was not significant as a whole (p=0.071), but it was significant in the subgroup analysis of 41-70% total body surface area burned. In that subgroup there were zero cases of death in the treatment group versus five cases in the control group (p=0.005). Our findings suggest that in acute burns, lactobacillus bacteria food additives may be clinically beneficial in patients with total burned body surface area of 41-70%. ( view less ) Shahar Lavi,Ori Nevo,Israel Thaler,Rimma Rosenfeld,Lior Dayan,Nir Hirshoren,Lior Gepstein,Giris Jacob Aging, independently from the hormonal status, is a major risk factor for cardiovascular morbidity in healthy women. Therefore, we studied the effect of healthy aging on the cardiovascular homeostatic mechanisms in premenopausal and postmenopausal women with similar estrogen levels. Twelve healthy ... ( view more )postmenopausal women, confirmed by follicular-stimulating hormone (FSH) and luteal hormone (LH) levels, were compared with 14 normally menstruating women during the early follicular phase (young-EF), to avoid as much as possible the effects of estrogen. Systolic BP was 108 +/- 1.5 vs. 123 +/- 2.5 (P < 0.001), supine norepinephrine was 260 +/- 30 vs. 216 +/- 45 and upright 640 +/- 100 vs. 395 +/- 50 pg/ml (P = 0.05) in young-EF vs. postmenopausal, respectively. Plasma renin activity and aldosterone remained unchanged. Vagal cardiac tone indices decreased significantly with aging (young-EF vs. postmenopausal): high-frequency (HF) band, root mean square successive differences (rMSSD) and proportion of R-R intervals >50 ms (PNN50%) were 620 +/- 140 vs. 270 +/- 70 (P = 0.04), 53 +/- 7 vs. 30 +/- 3 (P = 0.02), and 23 +/- 5 vs. 10 +/- 3 (P = 0.04), respectively. LF to HF ratio was 0.85 +/- 0.17 in young-EF and became 1.5 +/- 0.22 in postmenopausal (P = 0.03). Both arms of the baroreflex, +BRS (29 +/- 5 vs. 13.5 +/- 2.5, P = 0.01) and -BRS (26 +/- 4 vs. 15 +/- 1.5, P = 0.02) decreased with aging. Cardiovascular alpha(1)-adrenoreceptor responsiveness significantly increased and beta-decreased in postmenopausal compared with young EF (P < 0.001, both). The corrected QT intervals (QTc) were similar, whereas corrected JT intervals (JTc) and JTc to QTc ratio were prolonged in the postmenopausal group. We conclude that in young women, parasympathetic control is the main regulator of the cardiovascular system and in postmenopausal women, sympathetic tone dominates. The transition from parasympathetic to sympathetic control may contribute to the increased cardiovascular morbidity with aging. ( view less ) Lior Weissman,Milana Fraiberg,Lior Shine,Jacob Garty,Ayala Hochman The aim of this study was to identify, in the lichen Ramalina lacera, antioxidants that could provide indications of air pollution stress, and respond earlier than traditionally used structural/physiological parameters. The pollution-sensitive lichen R. lacera was transplanted from its relatively u... ( view more )npolluted natural habitat to two air-polluted sites for a period of up to 6 months. The superoxide dismutase and catalase activities, total water- and lipid-soluble low-molecular-weight antioxidant capacities and chlorophyll b/chlorophyll a ratios were assessed every 6 weeks. The earliest signs of oxidative stress were detected in the activities of fungal copper/zinc-superoxide dismutase, algal iron-superoxide dismutase and water-soluble low-molecular-weight antioxidants, which increased significantly as early as 42 days after exposure to pollution. Catalase activity increased in lichens transplanted to the polluted sites after 90 days. All activities decreased towards the end of the experiment. The impact of air pollution on R. lacera, using the traditionally employed parameter of chlorophyll b/chlorophyll a ratio, was only detected after 6 months of exposure to air pollution. Our results indicate that antioxidant parameters may serve as improved early-warning indicators of air pollution stress in lichens. ( view less ) Lior Yankelson,Lior Gepstein Gene therapy, cell therapy, and tissue engineering are emerging as novel experimental therapeutic paradigms for a variety of cardiovascular disorders. In the current report we will review the possible implications of these emerging technologies in the field of cardiac electrophysiology. Initially, ... ( view more )the possible role of myocardial gene and cell therapies in creating a biological alternative to electronic pacemakers for the treatment of bradyarrhythmias will be discussed. This will be followed by a description of the possible applications of using similar strategies for the treatment of common tachyarrhythmias. Finally, the electrophysiological implications of cardiac stem cell therapy for heart failure, as well as the possible in vitro applications of stem cell technology for electrophysiological studies and drug screening, will be discussed. While these emerging strategies provide a paradigm shift from conventional treatment modalities, this field is still at its infancy and several obstacles, discussed in this review, should be overcome before any clinical breakthroughs can be expected. ( view less ) Izhar Karbat,Lior Cohen,Nicholas Gilles,Dalia Gordon,Michael Gurevitz,Karbat Izhar,Cohen Lior,Gilles Nicholas,Gordon Dalia,Gurevitz Michael Gating modifiers constitute a large group of polypeptide toxins that interact with the voltage-sensing module of ion channels. Among them, scorpion beta-toxins induce a negative shift in the voltage dependence of sodium channel activation. To explain their effect, a "voltage sensor trapping" model ... ( view more )has been proposed in which the voltage sensor of domain-II (DIIS4) is trapped in an outward, activated position by a prebound beta-toxin upon membrane depolarization. Whereas toxin effect on channel activation was enhanced upon neutralization of the two outermost arginines in DIIS4, toxin residues involved in sensor trapping have not been identified. Using the scorpion excitatory beta-toxin, Bj-xtrIT, we found two conserved acidic residues, Glu15 and Glu30, mandatory for toxin action. Whereas mutagenesis of Glu30 affected both toxicity and binding affinity, substitutions E15A/F abolished activity but had minor effects on binding. Complete uncoupling of activity from binding was obtained with mutant E15R, acting as an efficient antagonist of Bj-xtrIT. On the basis of the voltage sensor trapping model and our results, we propose that Glu15 interacts with the emerging gating charges of DIIS4 upon membrane depolarization. Conserved acidic residues found in a variety of gating modifiers from scorpions and spiders may interact similarly with the voltage sensor. ( view less ) B Bourke,P M Sherman,D Woodward,H Lior,V L ChanTo determine the genomic relatedness among a selection of animal and human Campylobacter upsaliensis isolates, macrorestriction profiles were generated for 20 C. upsaliensis strains, among 7 serogroups, using pulsed-field gel electrophoresis (PFGE). XhoI, SalI and SacII restriction enzyme profiles ... ( view more )indicated genomic heterogeneity among strains. Using XhoI and SacII restriction enzyme digestion, genomic similarities between some pairs of strains were Lior serogroup specific. The genomic sizes of these isolates varied from 1.74 to 2.09 Mb. These results demonstrate molecular heterogeneity of this species similar to that found among Helicobacter pylori isolates. Among C. upsaliensis strains, PFGE is highly discriminatory and should prove a useful molecular typing method for epidemiological purposes. ( view less ) P Vandamme,P Pugina,G Benzi,R Van Etterijck,L Vlaes,K Kersters,J P Butzler,H Lior,S Lauwers In the autumn of 1983, an outbreak of recurrent abdominal cramps occurred in a nursery and primary school in the Rovigo area in Italy. None of the 10 affected children had diarrhea. An atypical Campylobacter-like organism was isolated from feces in all cases. Conventional enteropathogens were searc... ( view more )hed for but not detected. The Campylobacter-like organism was identified as Arcobacter butzleri by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins and cellular fatty acid analysis. Its identity was confirmed by DNA-DNA hybridizations versus Arcobacter reference strains. All of the preserved outbreak strains have identical protein profiles and phenotypic characteristics and belong to serogroup 1 of the Lior serotyping scheme on the basis of slide agglutination of crude and absorbed antisera of A. butzleri reference strains versus heat-labile antigens of live bacteria. These data point to an epidemiological relationship. The successive timing of the cases suggests person-to-person transmission. ( view less ) R K Bhadra,H Lior,S K Misra,S C Pal,G B NairC. jejuni and C. coli isolates from diverse sources in Calcutta were serotyped and biotyped according to the Lior scheme. Of the 55 strains examined, 85.5 per cent reacted with one or another of the 73 antisera available. This included the formation of two new serogroups, LIO 67 and LIO 76. C. coli... ( view more ) serogroup LIO 46 biotype II was the most frequently encountered strains (14.5%), followed by C. coli serogroup LIO 29, 55 biotype II (10.5%) and C. jejuni serogroup LIO 54, biotype I (5.5%). Serogroups recovered from animals and birds were also found to be prevalent in strains isolated from clinical sources, confirming the zoonotic implications of the disease. ( view less ) A M Hirschl,H Lior,D Wolf,G Stanek,M L Rotter,L Wende,H Flamm During the 1982-1986 period of all bacterial pathogens found to have caused diarrhoea, 35% belonged to the genus Campylobacter (C). Approximately 70% of the strains were isolated from persons under the age of 30 years, with a distinct peak of occurrence in the autumn. Biotyping and serotyping accor... ( view more )ding to Lior yielded the following results: C. jejuni biotype I: 32.9%, C. jejuni biotype II: 48.6%, C. coli biotype I: 10.3%, C. coli biotype II: 8.2%. From the 121 strains serotyped, 118 (97.5%) were typable. The serotypes most frequently encountered were type 1 (15.7%), 4 (9.9%), 2 and 11 (7.4% each). There were 2 familial outbreaks of Campylobacter enteritis which could be completely elucidated by biotyping and serotyping. One outbreak was caused by C. jejuni biotype I serotype 11, the other by C. jejuni biotype II serotype 6. Considering the frequent occurrence of Campylobacter infections, isolates should be routinely typed. The existing typing methods and schemes are highly developed. ( view less ) B W Brooks,M M Garcia,D E Fraser,H Lior,R B Stewart,A M LammerdingIn a recent meat survey, 10 of 13 (77%) Campylobacter coli isolates were susceptible to cephalothin. These organisms were isolated from nine slaughter cattle from eight meat packing establishments. All 10 isolates grew at 43 degrees C but not at 25 degrees C, were catalase and oxidase positive, and... ( view more ) were susceptible to nalidixic acid (30 micrograms) and cephalothin (30 micrograms). The cultures were subsequently identified as C. coli serogroup 20, biotype I (Lior scheme). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that protein and lipopolysaccharide profiles of whole cell preparations of the 10 cephalothin-susceptible strains and the reference strain for C. coli serogroup 20 were very similar. The plasmid profiles of these 11 strains were identical. ( view less ) M A Karmali,B Norrish,H Lior,B Heyes,A Monteath,H Montgomery During a five-day period, four neonates in a neonatal nursery developed Campylobacter entercolitis. Investigations suggested that cross-infection or common-source infection were unlikely and that the neonates acquired their infection during delivery from their respective mothers, three of whom were... ( view more ) also found to harbour Campylobacter jejuni in their stools. This suggestion was confirmed with use of the Lior serotyping system in a blind fashion. Each neonate was infected with a different serotype, and each of the three culture-positive mothers had the same serotype as her neonate. Examination of multiple colonies from the stools of five individuals showed that each was likely to have been infected by only one serotype. The presenting clinical features in the four neonates provides further evidence that neonatal Campylobacter entercolitis typically manifests as a benign, self-limited, nonfebrile, diarrheal illness with bloody stools. ( view less ) V Rusu,H Lior,S Lucinescu,M Kovacs The distribution per serotypes of 401 Campylobacter strains was studied by Lior's procedure, 241 strains being identified as C. jejuni (60.1%) and 160 as C. coli (39.9%): out of these 329 were of human origin (82%), including 225 C. jejuni and 104 C. coli, and 72 were isolated from fowls and swine ... ( view more )(16 C. jejuni, 56 C. coli). Out of the total of 401 strains, 363 (90.5%) were typable. Among human strains, 32 serotypes were identified, more frequently encountered being (in a decreasing order) 29, 4, 21, 2, 1, 9, 59, 44, 36, 48, 28, 8, 47, 5, 11, 57, 10, 32, 55 which sum up 89.5% of the 296 typable strains. Within C. jejuni species, serogroup 4 was the predominant one and for the C. coli one--29. Animal strains were distributed into 16 serogroups out of which 29, 26, 48, 55, 44 were more frequently encountered. The serogroups identified among animal strains were encountered, with a single exception, among human strains, too, but with a different frequency. The importance of serogrouping as an epidemiological marker, was confirmed by serogroup identity of the strains within some family foci, by the oneness, as serogroups, in the case of repeated isolations from the same patient as well as by establishing an epidemiological diagnosis, in the case of an epidemic, occurring among the members of a given community. ( view less ) M Millson,M Bokhout,J Carlson,L Spielberg,R Aldis,A Borczyk,H Lior A large outbreak of Campylobacter jujuni gastroenteritis attributed to contamination of an unchlorinated municipal water system was investigated. Unlike most previous summer outbreaks, this one began in early spring and was attributed to meltwater entering one or more municipal wells. 241 suspected... ( view more ) cases were documented, but retrospective information from local health care workers suggested a much larger outbreak. 45 laboratory-confirmed cases participated in a case-control study which showed a significant association between infection and amount of town water consumed. Stool specimens from 29 patients were studied with detailed serotyping by the method of Lior, with eight known serotypes and one previously unknown one identified. It is concluded that intensive surveillance of water quality during periods of spring runoff is essential, and that timely reporting of disease outbreak patterns in emergency department settings is necessary to protect the public's health. ( view less ) Jenq-Lin Yang,Lior Weissman,Vilhelm A Bohr,Mark P Mattson By producing ATP and regulating intracellular calcium levels, mitochondria are vital for the function and survival of neurons. Oxidative stress and damage to mitochondrial DNA during the aging process can impair mitochondrial energy metabolism and ion homeostasis in neurons, thereby rendering them ... ( view more )vulnerable to degeneration. Mitochondrial abnormalities have been documented in all of the major neurodegenerative disorders-Alzheimer's, Parkinson's and Huntington's diseases, and amyotrophic lateral sclerosis. Mitochondrial DNA damage and dysfunction may be downstream of primary disease processes such as accumulation of pathogenic proteins. However, recent experimental evidence demonstrates that mitochondrial DNA damage responses play important roles in aging and in the pathogenesis of neurodegenerative diseases. Therapeutic interventions that target mitochondrial regulatory systems have been shown effective in cell culture and animal models, but their efficacy in humans remains to be established. ( view less ) Lior Mayo,Jasmine Jacob-Hirsch,Ninette Amariglio,Gideon Rechavi,Marie-Jo Moutin,Frances E Lund,Reuven Stein Microglia, the resident immune cells of the CNS, are normally quiescent but become activated after infection or injury. Their properties then change, and they promote both repair and damage processes. The extent of microglial activation is regulated, in part, by activation-induced cell death (AICD)... ( view more ). Although many apoptotic aspects of the microglial AICD mechanism have been elucidated, little is known about the connection between the activation step and the death process. Using mouse primary microglial cultures, we show that the ectoenzyme CD38, via its calcium-mobilizing metabolite cyclic-ADP-ribose (cADPR), helps promote microglial activation and AICD induced by LPS plus IFN-gamma (LPS/IFN-gamma), suggesting that CD38 links the two processes. Accordingly, CD38 expression and activity, as well as the intracellular calcium concentration ([Ca(2+)](i)) in the primary microglia were increased by LPS/IFN-gamma treatment. Moreover, CD38 deficiency or treatment with cADPR antagonists conferred partial resistance to LPS/IFN-gamma-induced AICD and also reduced [Ca(2+)](i). Microglial activation, indicated by induced expression of NO synthase-2 mRNA and production of NO, secretion and mRNA expression of TNF-alpha and IL-12 p40, and expression of IL-6 mRNA, was attenuated by CD38 deficiency or cADPR-antagonist treatment. The observed effects of CD38 on microglial activation are probably mediated via a cADPR-dependent increase in [Ca(2+)](i) and the effect on AICD by regulation of NO production. Our results thus suggest that CD38 significantly affects regulation of the amount and function of activated microglia, with important consequences for injury and repair processes in the brain. ( view less ) Nilly Shimony,Idit Avrahami,Raphael Gorodetsky,Gregory Elkin,Keren Tzukert,Lior Zangi,Lilia Levdansky,Lina Krasny,Yosef S Haviv BACKGROUND: In epithelial and endothelial cells, detachment from the matrix results in anoikis, a form of apoptosis, whereas stromal and cancer cells are often anchorage independent. The classical anoikis model is based on static 3D epithelial cell culture conditions (STCK). METHODS: We characteriz... ( view more )ed a new model of renal, stromal and mesenchymal stem cell (MSC) matrix deprivation, based on slow rotation cell culture conditions (ROCK). This model induces anoikis using a low shear stress, laminar flow. The mechanism of cell death was determined via FACS (fluorescence-activated cell sorting) analysis for annexin V and propidium iodide uptake and via DNA laddering. RESULTS: While only renal epithelial cells progressively died in STCK, the ROCK model could induce apoptosis in stromal and transformed cells; cell survival decreased in ROCK versus STCK to 40%, 52%, 62% and 7% in human fibroblast, rat MSC, renal cell carcinoma (RCC) and human melanoma cell lines, respectively. Furthermore, while ROCK induced primarily apoptosis in renal epithelial cells, necrosis was more prevalent in transformed and cancer cells [necrosis/apoptosis ratio of 72.7% in CaKi-1 RCC cells versus 4.3% in MDCK (Madin-Darby canine kidney) cells]. The ROCK-mediated shift to necrosis in RCC cells was further accentuated 3.4-fold by H(2)O(2)-mediated oxidative stress while in adherent HK-2 renal epithelial cells, oxidative stress enhanced apoptosis. ROCK conditions could also unveil a similar pattern in the LZ100 rat MSC line where in ROCK 44% less apoptosis was observed versus STCK and 45% less apoptosis versus monolayer conditions. Apoptosis in response to oxidative stress was also attenuated in the rat MSC line in ROCK, thereby highlighting rat MSC transformation. CONCLUSIONS: The ROCK matrix-deficiency cell culture model may provide a valuable insight into the mechanism of renal and MSC cell death in response to matrix deprivation. ( view less ) Lior Lowenstein,Kimberly Kenton,Mary Pat FitzGerald,Linda Brubaker OBJECTIVE: The purposes of this study were to assess: 1) clinically relevant relationships between urinary diary and quality of life, and 2) reproducibility of validated questionnaires and urinary diaries in women with mixed urinary incontinence symptoms (MUI). STUDY DESIGN: Forty-seven women with ... ( view more )MUI completed 7-day diaries, the Urinary Distress Inventory (UDI-6), Incontinence Impact Questionnaire, and Medical, Epidemiological, and Social Aspects of Aging questionnaire 2 weeks apart. RESULTS: The number of urge incontinence episodes predicted incontinence severity on UDI-6 (R(2) = .38, P < .03). Except for the number of stress incontinence episodes, diary variables and questionnaire responses were reproducible (range from Spearman's rho = .7 to rho = .96, P < .001). CONCLUSION: The 6 questions of the UDI-6 adequately represent incontinence severity. With the exception of the number of stress incontinence episodes recorded on the 7-day diary, common incontinence measures are reproducible over 2 weeks. ( view less ) Lior Lowenstein,Mary P FitzGerald,Kimberly Kenton,Linda Brubaker,Ilan Gruenwald,Irena Papier,Ramón A Durazo-Arvizu,Christina Elliot,Elizabeth R Mueller,Yoram Vardi OBJECTIVE: The purpose of this study was to test the feasibility and validity of a continuous measurement of urinary sensation during cystometry. STUDY DESIGN: Subjects continuously recorded their level of urinary sensation during cystometry with the use of urodynamic diagnoses and responses to the... ( view more ) Medical Epidemiologic and Social Aspects of Aging (MESA) and Urinary Distress Inventory (UDI) scales. Trends in urinary sensation recordings by increasing percentage of maximum cystometric capacity (MCC) were captured and compared with the use of growth curves. RESULTS: Fifty-one women participated: 18 patients had detrusor overactivity incontinence (DOI); 15 patients had urodynamic stress incontinence; 9 patients had mixed incontinence, and 9 patients did not demonstrate incontinence. In the volume range between 35% and 75% of MCC, the mean sensation level was higher in the DOI group than the other groups (P < .04). Urge sensation at 50% of MCC correlated with UDI and MESA urge subscales (rho = 0.34, p < .03 and rho = 0.39, p < .02). CONCLUSION: This study demonstrates that the Urgeometer measurement is feasible and correlates with urodynamic diagnosis and the severity and bother from urge incontinence. ( view less )
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