bluuebunny/crossref_metadata_2025 huggingface dataset:
Delayed restoration of Mg2+content and transport in liver cells following ethanol withdrawal
Authors: Torres Lisa M. , Cefaratti Christie , Berti-Mattera Liliana , Romani Andrea
Published: June 2009

Liver cells from rats chronically fed a Lieber-De Carli diet for 3 wk presented a marked decreased in tissue Mg2+content and an inability to extrude Mg2+into the extracellular compartment upon stimulation with catecholamine, isoproterenol, or cell-permeant cAMP analogs. This defect in Mg2+extrusion was observed in both intact cells and purified liver plasma membrane vesicles. Inhibition of adrenergic or cAMP-mediated Mg2+extrusion was also observed in freshly isolated hepatocytes from control rats incubated acutely in vitro with varying doses of ethanol (EtOH) for 8 min. In this model, however, the defect in Mg2+extrusion was observed in intact cells but not in plasma membrane vesicles. In the chronic model, upon removal of EtOH from the diet hepatic Mg2+content and extrusion required ∼10 days to return to normal level both in isolated cells and plasma membrane vesicles. In hepatocytes acutely treated with EtOH for 8 min, more than 60 min were necessary for Mg2+content and extrusion to recover and return to the level observed in EtOH-untreated cells. Taken together, these data suggest that in the acute model the defect in Mg2+extrusion is the result of a limited refilling of the cellular compartment(s) from which Mg2+is mobilized upon adrenergic stimulation rather than a mere defect in adrenergic cellular signaling. The chronic EtOH model, instead, presents a transient but selective defect of the Mg2+extrusion mechanisms in addition to the limited refilling of the cellular compartments.
Parent-of-origin effects on cardiac response to pressure overload in mice
Authors: Barrick Cordelia J. , Dong Anping , Waikel Rebekah , Corn Drew , Yang Fanmuyi , Threadgill David W. , Smyth Susan S.
Published: June 2009

Left ventricular (LV) hypertrophy (LVH) is an independent risk factor for cardiovascular mortality and is commonly caused by hypertension. In rodents, transverse aortic constriction (TAC) is a model regularly employed in mechanistic studies of the response of the LV to pressure overload. We previously reported that inbred strains of male mice manifest different cardiac responses to TAC, with C57BL/6J (B6) developing LV dilatation and impaired contractility and 129S1/SvImJ (129) males displaying concentric LVH. In the present study, we investigated sex and parent-of-origin effects on the response to TAC by comparing cardiac function, organ weights, expression of cardiac hypertrophy markers, and histology in female B6 and female 129 mice and in F1 progeny of reciprocal crosses between B6 and 129 mice (B6129F1 and 129B6F1). Five weeks after TAC, heart weight increased to the greatest extent in 129B6F1 mice and the least extent in 129 and B6129F1 mice. Female 129B6F1 and B6 mice were relatively protected from the increase in heart weight that occurs in their male counterparts with pressure overload. The response to TAC in 129 consomic mice bearing the B6 Y chromosome resembled that of 129 rather than 129B6F1 mice, indicating that the B6 Y chromosome does not account for the differences in the reciprocal cross. Our results suggest that susceptibility to LVH is more complex than simple Mendelian inheritance and that parental origin effects strongly impact the LV response to TAC in these commonly used inbred strains.
Improving assessment of daily energy expenditure by identifying types of physical activity with a single accelerometer
Authors: Bonomi A. G. , Plasqui G. , Goris A. H. C. , Westerterp K. R.
Published: June 2009

Accelerometers are often used to quantify the acceleration of the body in arbitrary units (counts) to measure physical activity (PA) and to estimate energy expenditure. The present study investigated whether the identification of types of PA with one accelerometer could improve the estimation of energy expenditure compared with activity counts. Total energy expenditure (TEE) of 15 subjects was measured with the use of double-labeled water. The physical activity level (PAL) was derived by dividing TEE by sleeping metabolic rate. Simultaneously, PA was measured with one accelerometer. Accelerometer output was processed to calculate activity counts per day (ACD) and to determine the daily duration of six types of common activities identified with a classification tree model. A daily metabolic value (METD) was calculated as mean of the MET compendium value of each activity type weighed by the daily duration. TEE was predicted by ACD and body weight and by ACD and fat-free mass, with a standard error of estimate (SEE) of 1.47 MJ/day, and 1.2 MJ/day, respectively. The replacement in these models of ACD with METD increased the explained variation in TEE by 9%, decreasing SEE by 0.14 MJ/day and 0.18 MJ/day, respectively. The correlation between PAL and METD ( R2 = 51%) was higher than that between PAL and ACD ( R2 = 46%). We conclude that identification of activity types combined with MET intensity values improves the assessment of energy expenditure compared with activity counts. Future studies could develop models to objectively assess activity type and intensity to further increase accuracy of the energy expenditure estimation.
In vivo oxidative capacity varies with muscle and training status in young adults
Authors: Larsen Ryan G. , Callahan Damien M. , Foulis Stephen A. , Kent-Braun Jane A.
Published: June 2009

It is well established that exercise training results in increased muscle oxidative capacity. Less is known about how oxidative capacities in distinct muscles, in the same individual, are affected by different levels of physical activity. We hypothesized that 1) trained individuals would have higher oxidative capacity than untrained individuals in both tibialis anterior (TA) and vastus lateralis (VL) and 2) oxidative capacity would be higher in TA than VL in untrained, but not in trained, individuals. Phosphorus magnetic resonance spectroscopy was used to measure the rate of phosphocreatine recovery ( kPCr), which reflects the rate of oxidative phosphorylation, following a maximal voluntary isometric contraction of the TA and VL in healthy untrained (7 women, 7 men, 25.7 ± 3.6 yr; mean ± SD) and trained (5 women, 7 men, 27.5 ± 3.4 yr) adults. Daily physical activity levels were measured using accelerometry. The trained group spent threefold more time (∼90 vs. ∼30 min/day; P < 0.001) in moderate to vigorous physical activity (MVPA). Overall, kPCrwas higher in VL than in TA ( P = 0.01) and higher in trained than in untrained participants ( P < 0.001). The relationship between kPCrand MVPA was more robust in VL ( r = 0.64, P = 0.001, n = 25) than in TA ( r = 0.38, P = 0.06, n = 25). These results indicate greater oxidative capacity in vivo in trained compared with untrained individuals in two distinct muscles of the lower limb and provide novel evidence of higher oxidative capacity in VL compared with TA in young humans, irrespective of training status. The basis for this difference is not known at this time but likely reflects a difference in usage patterns between the muscles.
Mechanical properties and collagen cross-linking of the patellar tendon in old and young men
Authors: Couppé C. , Hansen P. , Kongsgaard M. , Kovanen V. , Suetta C. , Aagaard P. , Kjær M. , Magnusson S. P.
Published: June 2009

Age-related loss in muscle mass and strength impairs daily life function in the elderly. However, it remains unknown whether tendon properties also deteriorate with age. Cross-linking of collagen molecules provides structural integrity to the tendon fibrils and has been shown to change with age in animals but has never been examined in humans in vivo. In this study, we examined the mechanical properties and pyridinoline and pentosidine cross-link and collagen concentrations of the patellar tendon in vivo in old (OM) and young men (YM). Seven OM (67 ± 3 years, 86 ± 10 kg) and 10 YM (27 ± 2 years, 81 ± 8 kg) with a similar physical activity level (OM 5 ± 6 h/wk, YM 5 ± 2 h/wk) were examined. MRI was used to assess whole tendon dimensions. Tendon mechanical properties were assessed with the use of simultaneous force and ultrasonographic measurements during ramped isometric contractions. Percutaneous tendon biopsies were taken and analyzed for hydroxylysyl pyridinoline (HP), lysyl pyridinoline (LP), pentosidine, and collagen concentrations. We found no significant differences in the dimensions or mechanical properties of the tendon between OM and YM. Collagen concentrations were lower in OM than in YM (0.49 ± 0.27 vs. 0.73 ± 0.14 mg/mg dry wt; P < 0.05). HP concentrations were higher in OM than in YM (898 ± 172 vs. 645 ± 183 mmol/mol; P < 0.05). LP concentrations were higher in OM than in YM (49 ± 38 vs. 16 ± 8 mmol/mol; P < 0.01), and pentosidine concentrations were higher in OM than in YM (73 ± 13 vs. 11 ± 2 mmol/mol; P < 0.01). These cross-sectional data raise the possibility that age may not appreciably influence the dimensions or mechanical properties of the human patellar tendon in vivo. Collagen concentration was reduced, whereas both enzymatic and nonenzymatic cross-linking of concentration was elevated in OM vs. in YM, which may be a mechanism to maintain the mechanical properties of tendon with aging.
Diurnal physiology: core principles with application to the pathogenesis, diagnosis, prevention, and treatment of myocardial hypertrophy and failure
Authors: Sole Michael J. , Martino Tami A.
Published: June 2009

The circadian system has been shown to be fundamentally important in human health and disease. Recently, there have been major advances in our understanding of daily rhythmicity, and its relevance to human physiology, and to the pathogenesis and treatment of cardiac hypertrophy and heart failure. Cardiovascular tissues, such as heart and blood vessels, show remarkable daily variation in gene expression, metabolism, growth, and remodeling. Moreover, synchrony of daily molecular and physiological rhythms is integral to healthy organ growth and renewal. Disruption of these rhythms adversely affects normal growth, also the remodeling mechanisms in disease, leading to gross abnormalities in heart and vessels. These observations provide new insights into the pathogenesis, diagnosis, treatment, and prevention of heart disease. In this review, we focus on the recent advances in circadian biology and cardiovascular function, with particular emphasis on how this applies to human myocardial hypertrophy and heart failure, and the implications and importance for translational medicine.
Sex hormone effects on body fluid and sodium regulation in women with and without exercise-associated hyponatremia
Authors: Stachenfeld Nina S. , Taylor Hugh S.
Published: June 2009

We hypothesized that exercise-associated hyponatremia (EAH) is a function of excess sodium loss combined with high water intake in women at risk for dysnatremias during endurance exercise. We further hypothesized that estradiol and progesterone exposure increases fluid retention and sodium loss during exercise in women at risk for EAH. For 16 days we suppressed estrogens and progesterone with a gonadotropin-releasing hormone antagonist (GnRH ant) in seven women with (Hypo) and nine women without (no Hypo) a history of hyponatremia; we added 17β-estradiol (0.2 mg/day patches) for days 4–16 (E2) and progesterone (200 mg/day) for days 13–16 (E2-P4). Under each hormone condition, subjects cycled in 35°C at 65% peak oxygen consumption (V̇o2peak) for 60 min, then at 55–60% V̇o2peak for 120 min. Subjects drank 8 ml/kg of water (and replenished urine volume) every 30 min over the final 120 min of exercise. S[Na+] fell by 4.3, 3.9, and 3.1 meq/l ( P < 0.05) with drinking during GnRH ant, E2, and E2-P4 in Hypo, with little fall in no Hypo. Under all conditions, combined urine and sweat sodium loss were similar between Hypo [−85.6 (SD 36.2), −86.4 (SD 39.2), and −112.0 (SD 30.0) meq] and no Hypo [−98.0 (SD 54.8), −80.9 (SD 57.6), and −105.1 (SD 46.4) meq, for GnRH, E2, and E2-P4], as was mass balance of electrolytes (EMB) for Hypo [−104.8 (SD 32.8), −103.6 (SD 42.1), and −132.8 (SD 34.9) meq] compared with no Hypo [−128.8 (SD 57.2), −113.5 (SD 61.1), and −143.4 (SD 49.6) meq for GnRH, E2, and E2-P4]. Mass balance of water [VMB, for Hypo, 0.42 (SD 0.10), 0.62 (SD 0.25), and −0.11 (SD 0.11) liter] compared with no Hypo [0.01 (SD 0.15), 0.03 (SD 17), and −0.16 (SD 0.13) liter for GnRH, E2, and E2-P4, P < 0.05] indicates water retention was the primary contributor to the lower S[Na+] in Hypo women.
Exercise training decreases the size and alters the composition of the neointima in a porcine model of percutaneous transluminal coronary angioplasty (PTCA)
Authors: Fleenor Bradley S. , Bowles Douglas K.
Published: June 2009

Exercise training (EX) following percutaneous transluminal coronary angiography (PTCA) reduces progression to restenosis and increases event-free survival rates. Our aim was to determine whether EX inhibits lesion development and/or alters the extracellular matrix (ECM) composition of the neointima (NI) in a porcine PTCA model. Miniature Yucatan swine were assigned to cage confinement (SED) or EX for 20 wk. After 16 wk, all animals underwent a PTCA procedure of the left anterior descending artery (LAD) and left circumflex artery (LCX), with subsequent placement of an externalized jugular catheter. Animals recovered for 2 days and then resumed the previous protocol of SED or EX. Twelve days following PTCA, all animals received an intravenous bromodeoxyuridine (BrdU) injection to label proliferating cells. At 28 days following PTCA, the animals were euthanized, the LAD and LCX excised, and underwent standard histological processing for total collagen, type I collagen, fibronectin, BrdU, and Verhoeff-van Gieson stain. Our results demonstrate that EX significantly decreased lesion size and NI proliferation (−48%) in the LAD ( P < 0.05) but not the LCX. Furthermore, EX attenuated type I collagen expression only in LAD, whereas total collagen was increased (5.9%) and fibronectin was decreased (−7.9%) in the NI of both vessels ( P < 0.05). In conclusion, EX following PTCA may increase event-free survival rates following PTCA by decreasing lesion size and altering ECM composition.
Heat stress increases myonuclear number and fiber size via satellite cell activation in rat regenerating soleus fibers
Authors: Oishi Yasuharu , Hayashida Mari , Tsukiashi Shinsuke , Taniguchi Kohachi , Kami Katsuya , Roy Roland R. , Ohira Yoshinobu
Published: June 2009

To investigate the effects of heat stress (hyperthermia) on muscle degeneration-regeneration, the soleus muscles of adult male Wistar rats were injected bilaterally with a single injection of bupivacaine. The rats were assigned to a sedentary control (Con), heat stress (Heat), bupivacaine-injected (BPVC), or bupivacaine-injected plus heat stress (BPVC+Heat) group. Heat stress was induced in the Heat and BPVC+Heat groups by immersion of the lower half of the body into water maintained at 42 ± 1°C for 30 min 48 h after the injection of bupivacaine and every other day during the following 1 or 2 wk. The soleus muscles in all groups were excised 24 h after the final bout of heat stress. Mean muscle weight, fiber cross-sectional area, myonuclear number, and heat shock protein 72 (Hsp72) and calcineurin protein levels were lower in the BPVC than in the Con or Heat groups at both time points. In contrast, several of these parameters in the BPVC+Heat group were not different or higher than in the Con or Heat groups at the 1- and/or 2-wk time points. The number of total and activated satellite cells, estimated by analyses of Pax7-negative, M-cadherin-negative, and MyoD-positive nuclei, was greater in BPVC+Heat than in all other groups. Combined, the results indicate that heat stress-related activation of satellite cells and upregulation of Hsp72 and calcineurin expression played important roles in the regeneration of the soleus fibers after bupivacaine injection.
The New Standard of Fluids for Hemodialysis in Japan
Authors: Kawanishi Hideki , Masakane Ikuto , Tomo Tadashi
Published: June 2009

The standard of fluids for hemodialysis is being evaluated by the International Organization for Standardization (ISO), and will be decided within a few years. In 2008, the Japanese Society for Dialysis Therapy (JSDT) proposed the standard of fluids for hemodialysis by taking the draft ISO standard into consideration and the circumstances in Japan. It was characteristically a standard for Japan, where the central dialysis fluid delivery system (CDDS) is routinely used. In addition, the therapeutic application of each dialysis fluid is clarified. Since high-performance dialyzers are frequently employed in Japan, the standard recommends that ultrapure dialysis fluid be used for all dialysis modalities at all dialysis facilities. It also recommends that the dialysis equipment safety management committee at each facility validate the microbiological qualities of online-prepared substitution fluid, making the responsibility of the dialysis facility clear. This standard is more rigid than those of other countries, and is expected to contribute to improvements in the survival outcome of dialysis patients.
Bacteriological Water Quality in the Central Dialysis Fluid Delivery System from the Survey of the Japanese Society for Dialysis Therapy
Authors: Masakane Ikuto , Takemoto Yoshiaki , Nakai Shigeru , Tsubakihara Yoshiharu , Akiba Takashi , Watanabe Yuzo , Iseki Kunitoshi
Published: June 2009

The Japanese Society for Dialysis Therapy (JSDT) surveyed all dialysis facilities for bacteriological quality of dialysis fluid and quality controls for dialysis fluid in 2006 and 2007. The JSDT collected the data for endotoxin (ET) levels, bacterial count and usage of ET retentive filters (ETRF). The JSDT standard for ET level in dialysis fluid (<0.050 EU/ml) was achieved in 89.0% in 2006 and in 93.6% in 2007. The JSDT standard for bacterial cell counts in dialysis fluid (<100 cfu/ml) was achieved in 96.9% in 2006 and in 97.4% in 2007. The central dialysis fluid delivery system (CDDS) is a unique system developed in Japan which has easy handling for daily maintenance of delivery systems, but it has been pointed out that CDDS has a weak point for the protection of biofilms. However, the bacteriological water qualities of dialysis fluid in CDDS were proven to be extremely high in most Japanese dialysis facilities by JSDT surveys. Bacteriological water quality has a strong impact on the patient outcome. The acceptable level of ET of dialysis fluid should be <0.1 EU/ml based on the results of JSDT survey. The excellent water quality in CDDS might be one of the important factors which help good patient survival in chronic dialysis in Japan.
Advances and Advantages in Recent Central Dialysis Fluid Delivery System
Authors: Koda Yutaka , Mineshima Michio
Published: June 2009

The central dialysis fluid delivery system (CDDS) has been working exclusively in Japan since 1970, a fact which proves that CDDS is useful in the multi-patient treatment with in-center regular hemodialysis. It is a cost-effective, labor-saving and time-tested system with 40 years of experience. Microbial safety is improved by a refined system design, multiple endotoxin retentive filters (ETRF) and daily overnight disinfection of the entire system. Consequently, the usage of backfiltered dialysate is now accepted by regulatory authorities in some bedside consoles and even more the creation of dialysate is possible for infusion to meet its requirements. In many countries, especially in Asia, the number of end-stage renal disease patients is rapidly increasing. CDDS will contribute to such emerging situations with its easy handling and economical advantages.
Purification of Dialysis Water in the Central Dialysis Fluid Delivery System in Japan: A Prospective Observation Study
Authors: Uchino Junji , Kawasaki Tadayuki
Published: June 2009

Whereas the main measure of dialysis fluid purity is endotoxin (ET) activity in Japan, it is the viability count in Western countries. Because of this difference, little information is available concerning dialysis fluid purity determined in terms of viability count in Japan. Under these circumstances a fact-finding investigation was planned and conducted concerning dialysis fluid purity to demonstrate the effectiveness of dialysis fluid purification measures. 93 medical institutions are equipped with the central dialysis fluid delivery system (CDDS) unique to Japan. Almost all medical institutions surveyed have achieved the purification level of ultrapure dialysis fluid after ETRF, but the methods of ETRF use and management widely vary with each institution so that early validation of the methods of evaluation of ET inhibition and system management is in urgent need. It is also important that simple universal microbial monitoring and purification procedures be diffused far and wide as suggested by the Purification Guidelines proposed by us.
SMC Proteins and Their Multiple Functions in Higher Plants
Authors: Schubert V.
Published: June 2009

In plants as in other eukaryotes, SMC (Structural Maintenance of Chromosome) protein complexes and proteins interacting with them are essential for sister chromatid cohesion, chromosome condensation, DNA repair and recombination. The presence of paralogous genes for various components of the different SMC complexes allowed diversification of their biological functions during evolution of higher plants. Here I summarize the multiple functions of plant SMC complexes and some of the particularities these proteins show in comparison to those of other organisms.
Identification of a Maize Neocentromere in an Oat-Maize Addition Line
Authors: Topp C.N. , Okagaki R.J. , Melo J.R. , Kynast R.G. , Phillips R.L. , Dawe R.K.
Published: June 2009

We report a neocentromere event on maize chromosome 3 that occurred due to chromosome breakage. The neocentromere lies on a fragment of the short arm that lacks the primary centromere DNA elements, CentC and CRM. It is transmitted in the genomic background of oat via a new centromere (and kinetochore), as shown by immunolocalization of the oat CENH3 protein. Despite normal transmission of the maize fragment in most progeny, neocentromeres appear to vary in size within the same tissue, as shown by fluorescent measurements. A secondary truncation in one line lowered mitotic transmission to 3% and precipitously reduced the size of the chromosome. The results support the view that neocentromere formation is generally associated with major genomic disturbances such as wide species crosses or deletion of an existing centromere. The data further suggest that new centromeres may undergo a period of instability that is corrected over a period of several generations.
Avian Lampbrush Chromosomes: a Powerful Tool for Exploration of Genome Expression
Authors: Gaginskaya E. , Kulikova T. , Krasikova A.
Published: June 2009

Lampbrush chromosomes (LBCs) are highly extended bivalents that function in the growing oocytes of many animals. Due to their distinctive chromomere-loop organization and intense transcriptional activity of lateral loops the LBCs, mainly amphibian ones, have served as a powerful system for exploring the general principles of chromosome organization and function. The exploitation of avian LBCs has considerably broadened the opportunities for comparative genome research and for cytogenetic analysis of domestic species. In this review we highlight the advantages of avian LBCs for research in different areas including integration of genome organization studies with studies on gene activity in vivo, analysis of co-transcriptional events occurring on nascent transcripts and investigation of chromosome-associated intranuclear domains. Recent findings concerning the organization of transcriptionally active and silent chromatin together with involvement of cohesin and condensin complexes into maintenance of structural integrity of LBCs are presented. The biological significance of the LBC phenomenon is discussed. The intensive transcription on LBCs shows some specific features: very long transcription units, deregulated termination, and transcription of non-coding satellite repeats. Here, based on the modern view on a role of RNA interference machinery in regulation of genome expression, we suggest a mechanism of initiation of satellite DNA transcription and offer a novel interpretation of the ‘classical’ hypothesis that sought to explain the significance of widespread transcription during oocyte growth.
Interaction Study of the Male Specific Lethal (MSL) Complex and Trans-Acting Dosage Effects in Metafemales of Drosophila melanogaster
Authors: Sun X. , Birchler J.A.
Published: June 2009

The effect of ectopic expression of male specific lethal 2 (msl2) on chromatin modification and gene expression was studied in Drosophila diploid females and metafemales (3X;2A). Results show that ectopic expression of MSL2 in transgenic msl2 females and metafemales sequesters the MOF histone acetylase to the X, which occurs concordantly with an increase of histone acetylation. Gene expression studies indicate that the X-linked genes are not affected by direct targeting of the MSL complex and the resulting increased H4Lys16 acetylation on the X chromosomes, suggesting one function of the MSL complex is to nullify the effect of a high level of histone acetylation. These results are not consistent with the hypothesis that the presence of the MSL complex conditions a two-fold upregulation. Autosomal gene expression is generally decreased in ectopically expressed MSL2 females, which correlates with the reduced autosomal histone acetylation. Metafemales show dosage compensation of X-linked genes with some autosomal reductions in expression. Interestingly, in metafemales with ectopically expressed MSL2, the autosomal expression is returned to a more normal level. There is a lower autosomal level of histone acetylation compared to the normal metafemales, suggesting a nullifying effect on the negative dosage effect of the X chromosome as previously hypothesized to occur in normal males.
Extrachromosomal Circular DNA in Eukaryotes: Possible Involvement in the Plasticity of Tandem Repeats
Authors: Cohen S. , Segal D.
Published: June 2009

Extrachromosomal circular DNA (eccDNA) is ubiquitous in eukaryotic organisms, and has been noted for more than 3 decades. eccDNA occurs in normal tissues and in cultured cells, is heterogeneous in size, consists of chromosomal sequences and reflects plasticity of the genome. Two-dimensional (2D) gel electrophoresis has been adapted for the detection and characterization of eccDNA. It shows that most eccDNA consists of chromosomal tandem repeats, both coding genes and satellite DNA and is organized as circular multimers of the repeating sequence. 2D gels were unable to detect dispersed repeats within the population of eccDNA. eccDNA, organized as circular multimers, can be formed de novo in Xenopus egg extracts, in the absence of DNA replication. These findings support a mechanism for the formation of eccDNA that involves intra-chromosomal homologous recombination between tandem repeats and looping-out. Furthermore, eccDNA appears to undergo extrachromosomal replication via a rolling circle mechanism. Hence, the formation of eccDNA from arrays of tandem repeats may cause deletions, and the possible re-integration of rolling-circle replication products could expand these arrays. This review summarizes recent experimental data which characterizes eccDNA in several organisms using 2D gel electrophoresis, and discusses its possible implications on the dynamics of chromosomal tandem repeats.
Role of Fluorescence in situ Hybridization in Sequencing the Tomato Genome
Authors: Stack S.M. , Royer S.M. , Shearer L.A. , Chang S.B. , Giovannoni J.J. , Westfall D.H. , White R.A. , Anderson L.K.
Published: June 2009

The tomato (Solanum lycopersicum L.) genome is being sequenced by a consortium of laboratories in 10 countries. Seventy-seven percent of the tomato genome (DNA) is located in repeat-rich, gene-poor, pericentric heterochromatin, while 23% of the genome is located in repeat-poor, gene-rich, distal euchromatin. It is estimated that approximately 90% of tomato’s nuclear genes can be characterized by limiting the sequencing effort to euchromatin while avoiding the problems involved in sequencing the repetitive DNA in heterochromatin. Sequencing is being performed on tomato nuclear DNA cloned into bacterial artificial chromosome (BAC) vectors. Fluorescence in situ hybridization (FISH) is used to help direct the sequencing effort by cytologically demonstrating the location of selected BACs on tomato chromosomes. While mitotic metaphase chromosomes are too short and compact for this purpose, long pachytene chromosomes are ideal. BACs localized in euchromatin can be used confidently as anchors for the assembly of BAC contigs that extend through the euchromatic length of each chromosome arm. Another important role for FISH is identification of BACs near telomeres and near borders with pericentric heterochromatin to indicate that sequencing should not extend much further. This role of FISH is enhanced by our ability to estimate base pair distances between localized BACs and these chromosomal features. Finally, it is noteworthy that when BAC-FISH is combined with chromosomal in situ suppression (CISS) hybridization to block repeats and localize single/low copy sequences, the great majority of BACs localize to single sites. This observation is consistent with tomato being an ancient diploid.
Spectral Analysis of Field Potential Recordings by Deep Brain Stimulation Electrode for Localization of Subthalamic Nucleus in Patients with Parkinson’s Disease
Authors: Miyagi Yasushi , Okamoto Tsuyoshi , Morioka Takato , Tobimatsu Shozo , Nakanishi Yoshitaka , Aihara Kazuyuki , Hashiguchi Kimiaki , Murakami Nobuya , Yoshida Fumiaki , Samura Kazuhiro , Nagata Shinji , Sasaki Tomio
Published: June 2009

Aims: Spectral analysis of local field potential (LFP) recorded by deep brain stimulation (DBS) electrode around the subthalamic nucleus (STN) in patients with Parkinson’s disease was performed. Methods: The borders of the STN were determined by microelectrode recording. The most eligible trajectory for the sensorimotor area of the STN was used for LFP recording while advancing the DBS electrode. Results: The low-frequency LFP power (θ- to β-band) increased from a few millimeters above the dorsal border of the STN defined by microelectrode recording; however, the low-frequency power kept the same level beyond the ventral border of the STN. Only high β-power showed close correlation to the dorsal and ventral borders of the STN. Conclusions: A spectral power analysis of LFP recording by DBS electrode helps with the final confirmation of the dorsal and ventral borders of the STN of Parkinson’s disease in DBS implantation surgery.
Prognostic Factors of Subthalamic Stimulation in Parkinson’s Disease: A Comparative Study between Short- and Long-Term Effects
Authors: Tsai Sheng-Tzung , Lin Sheng-Huang , Chou Yu-Cheng , Pan Yan-Hong , Hung Hsiang-Yi , Li Chi-Wei , Lin Shinn-Zong , Chen Shin-Yuan
Published: June 2009

Background/Aims: Bilateral subthalamic nucleus deep brain stimulation (STN-DBS) has been shown to have long-term benefits in Parkinson’s disease (PD). Through analyzing different variables, this study identified prognostic factors for the short- and long-term effects of STN-DBS. Methods: Thirty-six PD patients underwent bilateral STN-DBS. Clinical evaluations were performed 1 month before and 3 months after surgery, with additional follow-up examinations for a mean of 31.3 months. Results: There was a trend for long-term STN-DBS-induced improvements in the Unified Parkinson’s Disease Rating Scale (UPDRS) part II and part III measures to be greater in younger patients. Preoperative levodopa responsiveness only led to consistent UPDRS part III improvement from STN-DBS at 3 months, and this predictive value did not exist in the long term. The preoperative levodopa response of tremor and axial symptoms in motor disability predicted long-term DBS effect only. Preoperative cognitive function positively correlated with postoperative improvement from DBS in UPDRS part III during long-term follow-up only. Conclusions: The prognostic factors for STN-DBS benefit were different for short- and long-term follow-ups. Good prognostic factors for long-term STN-DBS for PD patients were good cognitive function and tremor dominance. Poor prognostic factors were related to older age and non-dopaminergic-responsive axial disability.
Current Oncological Treatment of Patients with Pancreatic Cancer in Germany: Results from a National Survey on behalf of the Arbeitsgemeinschaft Internistische Onkologie and the Chirurgische Arbeitsgemeinschaft Onkologie of the Germany Cancer Society
Authors: Boeck Stefan , Bruns Christiane J. , Sargent Mirja , Schäfer Claus , Seufferlein Thomas , Jauch Karl-Walter , Heinemann Volker
Published: June 2009

Background: No data have previously been available regarding the current treatment of patients with pancreatic cancer (PC) in German hospitals and medical practices. Methods: Between February 2007 and March 2008 we conducted a national survey [on behalf of the Arbeitsgemeinschaft Internistische Onkologie (AIO) and the Chirurgische Arbeitsgemeinschaft Onkologie (CAO)] regarding the current surgical and oncological treatment of PC in Germany. Standardized questionnaires were sent via mailing lists to members of the AIO and CAO (n = 1,130). The data were analyzed using SPSS software (version 16.0). Pre-defined subgroup analysis was performed by grouping the results of each question with regard to the professional site of the responding physician and to the number of patients treated in their institution by year. Results: 181 (16%) of the oncological questionnaires were sent back. For 61% of the participating centers, a histological confirmation of PC diagnosis is obligatory. 21% of physicians offer neoadjuvant therapy to patients with potentially resectable PC. In the adjuvant treatment after curative-intent surgery, gemcitabine (Gem) is regarded as standard of care by 71% after R0 resection and 62% after R1 resection. For patients with locally advanced PC, 52% of the participating centers recommend systemic chemotherapy, 17% prefer combined primary chemoradiotherapy. Most centers (59%) base their decision of combination regimens for metastatic disease on the performance status of their patients. In patients with a good status, 28% apply single-agent Gem, 3% use Gem + capecitabine, 12% Gem + erlotinib, 16% Gem + oxaliplatin, and 8% Gem + cisplatin. Only 28% of the survey doctors offer second-line treatment to the majority of their patients with advanced PC. Conclusion: Not every PC patient in Germany is treated according to the present S3 guidelines. Diagnosis and treatment of PC in Germany still need to be improved.
Podoplanin Expression Identified in Stromal Fibroblasts as a Favorable Prognostic Marker in Patients with Colorectal Carcinoma
Authors: Yamanashi Takahiro , Nakanishi Yukihiro , Fujii Gen , Akishima-Fukasawa Yuri , Moriya Yoshihiro , Kanai Yae , Watanabe Masahiko , Hirohashi Setsuo
Published: June 2009

Objective: The microenvironment of cancer plays a critical role in its progression. However, the molecular features of cancer-associated fibroblasts (CAFs) are less well understood than those of cancer cells. We investigated the clinicopathological significance of podoplanin expression in stromal fibroblasts in patients with colorectal cancer (CRC). Methods: We selected podoplanin as an upregulated marker in CAF from a DNA microarray experiment. Consequently, podoplanin was identified as an upregulated gene. Immunohistochemical podoplanin expression was investigated at the National Cancer Center Hospital, Tokyo, Japan, in 120 patients with advanced CRC, and its clinicopathological significance was examined. The biological function of podoplanin expression was also assessed by a coculture invasion assay with CRC cell lines such as HCT116 and HCT15. Results: Podoplanin expression was exclusively confined to stromal fibroblasts and absent in tumor cells. Podoplanin is absent in normal stroma except for lymphatic vessels. Staining was considered positive when over 30% of the cancer stroma was stained. Positive podoplanin expression was significantly correlated with a more distal tumor localization (p = 0.013) and a shallower depth of tumor invasion (p = 0.011). Univariate analysis revealed that negative podoplanin expression in stromal fibroblasts was significantly associated with reduced disease-specific survival (p = 0.0017) and disease-free survival (p < 0.0001). Multivariate analysis revealed that negative podoplanin expression (p = 0.016) and lymph node metastasis (p = 0.027) were significantly associated with disease-free survival. CRC cell invasion was augmented by co-culture with CAFs that were treated with siRNA for podoplanin. Conclusions: Our results suggest that a positive podoplanin expression in stromal fibroblasts could have a protective role against CRC cell invasion and is a significant indicator of a good prognosis in patients with advanced CRC, supported by biological analysis showing that podoplanin expression in CAFs is associated with decreased CRC cell invasion.
Phase I Study of Prolonged-Infusion Gemcitabine Combined with Cyclophosphamide in Patients with Metastatic Carcinoma of the Breast: Tolerability of an Optimal Dose Schedule
Authors: Estephan Fadi , Valero Vicente , Esteva Francisco J. , Mejia Jaime A. , Frye Debra K. , Ibrahim Nuhad K.
Published: June 2009

Background: A phase I study was initiated to determine the maximum tolerated dose (MTD) of prolonged-infusion gemcitabine combined with cyclophosphamide in patients with metastatic breast carcinoma (MBC). Methods: Patients with MBC were treated with gemcitabine infusion at 10 mg/m2/min and cyclophosphamide by intravenous piggyback injection, 4 h after initiation of the infusion. We treated 3–6 patients at a particular dose level until the MTD was determined. Results: Overall, 44 patients received a total of 197 courses of therapy. Both drugs were given on days 1, 8 and 15 to 14 patients (68 courses). Delayed white blood cell recovery necessitated first protocol amendment to drop cyclophosphamide on days 8 and 15 in 9 patients (43 cycles). A second amendment was needed to drop gemcitabine on day 15 because of thrombocytopenia in 21 patients (86 courses). The dose-limiting toxicity was thrombocytopenia. The MTD of an optimal dose schedule was 800 mg/m2 gemcitabine infused at a rate of 10 mg/m2/min on days 1 and 8, and 400 mg/m2 cyclophosphamide, by intravenous piggyback injection, on day 1, 4 h after initiation of the gemcitabine infusion. Conclusions: The MTD can be given safely every 4 weeks to patients with MBC. Phase II studies are warranted to evaluate the clinical activity of this therapy.
S-1 Monotherapy in Patients with Advanced Biliary Tract Cancer
Authors: Sasaki Takashi , Isayama Hiroyuki , Yashima Yoko , Yagioka Hiroshi , Kogure Hirofumi , Arizumi Toshihiko , Togawa Osamu , Matsubara Saburo , Ito Yukiko , Nakai Yousuke , Sasahira Naoki , Hirano Kenji , Tsujino Takeshi , Tada Minoru , Kawabe Takao , Omata Masao
Published: June 2009

Objective: The aim of this study was to evaluate the efficacy and safety of single-agent S-1 in patients with advanced biliary tract cancer. Methods: S-1 was administered orally at a dose of 80 mg/m2 for 28 days, followed by 14 days of rest (1 cycle); treatment was repeated until disease progression, unacceptable toxicity, or patient refusal. Results: Forty-five patients were enrolled between January 2005 and June 2008. Among these, 29 patients received S-1 as first-line chemotherapy and 16 patients received S-1 as second-line chemotherapy. The response rates for first- and second-line chemotherapy were 17.2 and 18.8%, respectively. The median times to progression for the first- and second-line chemotherapy groups were 4.2 and 5.5 months (p = 0.91), respectively. The median overall survival and 1-year survival rate for each group were 8.7 and 8.0 months and 42.2 and 38.2%, respectively (p = 0.62). Only the first-line chemotherapy group experienced grade 3/4 toxicities, including leukopenia (6.9%), neutropenia (10.3%), anemia (6.9%), thrombocytopenia (10.3%) and total bilirubin elevation (3.4%). Conclusion: S-1 monotherapy is a feasible and moderately efficacious treatment for advanced biliary tract cancer, as a first- or second-line chemotherapy regimen.
Evaluation of the Hemodynamic Status of Focal Hepatic Lesions 20 mm or Less in Diameter by Contrast-Enhanced Ultrasonography Using Sonazoid
Authors: Shiozawa Kazue , Watanabe Manabu , Sumino Yasukiyo
Published: June 2009

Objective: To clarify the validity of the evaluation of the hemodynamics of hepatic lesions ≤20 mm in diameter using contrast-enhanced ultrasonography (CE-US) with Sonazoid. Methods: Sixty-two hepatic lesions in 55 patients with chronic hepatitis or cirrhosis due to hepatitis C virus were studied. We evaluated by quantitative analysis the time intensity curve (TIC) on hepatic lesion and parenchyma in the early vascular phase and Kupffer imaging in the post-vascular phase. Results: TIC patterns were classified into those with a maximum slope (Max slope) steeper in the hepatic lesion than in the parenchyma (Pattern I), those with a Max slope similar in the hepatic lesion and parenchyma (Pattern II), and those with a Max slope gentler in the hepatic lesion than in the parenchyma (Pattern III). The blood flow was considered to be higher, the blood flow velocity to be faster, and the contrast agent to reach the lesion more rapidly in Pattern I lesions than in the hepatic parenchyma. Pattern III lesions showed that the velocity of arterial blood influx was slow. Conclusion: Our study suggested the possibility that the TIC allows a detailed evaluation of the hemodynamics of hepatic lesions.
Intraocular Pressure-Lowering Activity of Topical Application of Aegle Marmelos Fruit Extract in Experimental Animal Models
Authors: Agarwal Renu , Gupta Suresh Kumar , Srivastava Sushma , Saxena Rohit , Agrawal S.S.
Published: June 2009

Aim: The present study was designed to evaluate the intraocular pressure (IOP)-lowering activity of topical application of the aqueous extract of Aegle marmelos fruit in experimental animal models. Materials and Methods: New Zealand white rabbits with normal and experimentally elevated IOP using water loading and steroid-induced models were included in this study. The IOP-lowering effect of A. marmelos fruit extract in rabbits with experimentally elevated IOP was also compared with that of timolol 0.25%. Results: In rabbits with normal IOP, the A. marmelos fruit extract at a concentration of 1% showed the maximum IOP-lowering effect with 22.81% reduction from baseline IOP. The maximum IOP reduction achieved in water loading and steroid-induced models with the same concentration of A. marmelos was 27.57 and 28.41% from baseline, respectively. The efficacy was comparable to that of timolol after 45 min of water loading in the water loading model, and during the first 2 h of treatment in the steroid-induced model. Conclusion: A. marmelos fruit extract showed significant IOP-lowering activity in experimental animal models.
In vitro Antagonism of Rabeprazole and Metronidazole upon Clinical Isolates of Helicobacter pylori
Authors: Soto Alejandra , García Apolinaria , González Carlos L.
Published: June 2009

Background: Rabeprazole inhibits Helicobacter pylori in vitro. This property could influence the antibacterial activity of metronidazole or amoxicillin. Our goal was to evaluate in vitrothe effect of rabeprazole upon the antibacterial activity of metronidazole and amoxicillin. Methods: The minimal inhibitory concentration (MIC) was determined by agar dilution upon 30 clinical isolates, and the antibacterial effect of the combined drugs was estimated by calculating the fractional inhibitory concentration index (FICI). Results: Nine strains (30%) were resistant to metronidazole (MIC 8–256 mg/l) while only 1 strain was resistant to amoxicillin (MIC 1.0 mg/l). Rabeprazole also inhibited H. pylori (MIC 0.125–1.0 mg/l). The association of metronidazole and rabeprazole showed synergism in 30% of the strains (FICI ≤0.25), but also antagonism in 23.3% of the clinical isolates (FICI ≥4.0). Mostly, amoxicillin-rabeprazole showed an effect of synergism (53.3%). Conclusion: The results presented suggest the need to keep in mind that failure in eradication therapies based on metronidazole and rabeprazole could be due to an antagonism of the drugs.
Predictors of Higher Blood Pressure in a Clinical Setting in Normotensive Children: A Prospective Study
Authors: Bohlke Maristela , Dullius Fernanda , Menezes Juliana , Cunha Tiago , Boemer Hayslan , Menezes Adriana , Caruso Paulo
Published: June 2009

This study was designed to assess the predictors of a higher blood pressure (BP) in a clinical setting in a prospective cohort of normotensive children recruited in an outpatient pediatric clinical setting in Southern Brazil. The study evaluated 443 children 3–12 years of age. If the mean value of BP was greater than the 95th percentile for age, sex and height in the medical setting, children were scheduled to other two evaluations at home. The systolic BP percentile in the clinical consultation was associated with BMI and birth weight. The diastolic BP in the clinical consultation was associated with birth weight, age, and BMI. Weight excess, low birth weight and younger age were associated with a higher BP in normotensive children seen in a clinical setting. To our knowledge, this is the first study to describe the association between lower birth weight and a greater BP response to medical consultation. Our finding suggests that the tendency to higher white-coat effect is determined, at least to some extent, in intrauterine life.
Effect of Lactococcus lactis CLFP 100 and Leuconostoc mesenteroides CLFP 196 on Aeromonas salmonicida Infection in Brown Trout (Salmo trutta)
Authors: Balcázar José Luis , Vendrell Daniel , de Blas Ignacio , Ruiz-Zarzuela Imanol , Múzquiz José Luis
Published: June 2009

Aeromonas salmonicida is the etiological agent of furunculosis in salmonid fish. This pathogen is important from an epizootic perspective because fish surviving an outbreak can remain lifelong asymptomatic carriers, serving as reservoirs of infection. As a result, the early detection and the control of infection are essential to prevent the spread of new furunculosis outbreaks. We have thus analyzed the effect of probiotic administration on the incidence of A. salmonicida in brown trout (Salmo trutta), that were subjected to temperature stress. Treatment with probiotic strains (Lactococcus lactis CLFP 100 and Leuconostoc mesenteroides CLFP 196) resulted in a higher survival rate after challenge, activation of phagocytic cells in the head kidney, and a lower rate of pathogen proliferation in the intestine as determined by real-time PCR.