bluuebunny/crossref_metadata_2025 huggingface dataset:
Jurassic geothermal landscapes and fossil ecosystems at San Agustín, Patagonia, Argentina
Authors: Guido Diego M. , Channing Alan , Campbell Kathleen A. , Zamuner Alba
Published: January 2010

An extensive, well-preserved, Late Jurassic (
c
. 150 Ma) geothermal system at San Agustín farm in the Deseado Massif, Patagonia, Argentina, is described. This deposit, along with others previously known from the same region, partially fills a considerable gap between Cenozoic and scattered Palaeozoic hot spring localities reported worldwide. The San Agustín deposit is novel because it represents a large (1.4 km
2
) and nearly complete geothermal landscape. Siliceous hot spring facies, both subaerial and subaqueous, are exposed side by side in their original spatial and geological context, set amongst intrusive rhyolite domes and fluviolacustrine sediments. The Jurassic hot springs have preserved an entire local ecosystem containing microbes, arthropods, gastropods and plants exhibiting Lagerstätten-style preservation. Plant preservation, in particular, ranges from decayed litter, to seedling sprouts, and to dense stands in life orientation with intact anatomy. The San Agustín deposit shares some ecological, taphonomic and sedimentological characteristics with modern hot springs. As it formed in a pre-angiosperm world, it is akin to the famous hot spring-related Devonian Rhynie cherts of Scotland. It differs in having excellent exposure, and thus will probably contribute to a better understanding of biosignal preservation in extreme environments in the geological record.
Fluid Mechanics of Microrheology
Authors: Squires Todd M. , Mason Thomas G.
Published: January 2010

In microrheology, the local and bulk mechanical properties of a complex fluid are extracted from the motion of probe particles embedded within it. In passive microrheology, particles are forced by thermal fluctuations and probe linear viscoelasticity, whereas active microrheology involves forcing probes externally and can be extended out of equilibrium to the nonlinear regime. Here we review the development, present state, and future directions of this field. We organize our review around the generalized Stokes-Einstein relation (GSER), which plays a central role in the interpretation of microrheology. By discussing the Stokes and Einstein components of the GSER individually, we identify the key assumptions that underpin each, and the consequences that occur when they are violated. We conclude with a discussion of two techniques—multiple particle-tracking and nonlinear microrheology—that have arisen to handle systems in which the GSER breaks down.
Stimulation of neuronal KATPchannels by cGMP-dependent protein kinase: involvement of ROS and 5-hydroxydecanoate-sensitive factors in signal transduction
Authors: Chai Yongping , Lin Yu-Fung
Published: January 2010

The ATP-sensitive potassium (KATP) channel couples intracellular metabolic state to membrane excitability. Recently, we demonstrated that neuronal KATPchannels are functionally enhanced by activation of a nitric oxide (NO)/cGMP/cGMP-dependent protein kinase (PKG) signaling cascade. In this study, we further investigated the intracellular mechanism underlying PKG stimulation of neuronal KATPchannels. By performing single-channel recordings in transfected HEK293 and neuroblastoma SH-SY5Y cells, we found that the increase of Kir6.2/SUR1 (i.e., the neuronal-type KATP) channel currents by PKG activation in cell-attached patches was diminished by 5-hydroxydecanoate (5-HD), an inhibitor of the putative mitochondrial KATPchannel; N-(2-mercaptopropionyl)glycine, a reactive oxygen species (ROS) scavenger, and catalase, a hydrogen peroxide (H2O2)-decomposing enzyme. These reagents also ablated NO-induced KATPchannel stimulation and prevented the shifts in the single-channel open- and closed-time distributions resulting from PKG activation and NO induction. Bath application of H2O2reproduced PKG stimulation of Kir6.2/SUR1 but did not activate tetrameric Kir6.2LRKR368/369/370/371AAAA channels. Moreover, neither the PKG activator nor exogenous H2O2was able to enhance the function of KATPchannels in the presence of Ca2+chelators and calmodulin antagonists, whereas the stimulatory effect of H2O2was unaffected by 5-HD. Altogether, in this report we provide novel evidence that activation of PKG stimulates neuronal KATPchannels by modulating intrinsic channel gating via a 5-HD-sensitive factor(s)/ROS/Ca2+/calmodulin signaling pathway that requires the presence of the SUR1 subunit. This signaling pathway may contribute to neuroprotection against ischemic injury and regulation of neuronal excitability and neurotransmitter release by modulating the function of neuronal KATPchannels.
Mitochondrial matrix K+flux independent of large-conductance Ca2+-activated K+channel opening
Authors: Aldakkak Mohammed , Stowe David F. , Cheng Qunli , Kwok Wai-Meng , Camara Amadou K. S.
Published: January 2010

Large-conductance Ca2+-activated K+channels (BKCa) in the inner mitochondrial membrane may play a role in protecting against cardiac ischemia-reperfusion injury. NS1619 (30 μM), an activator of BKCachannels, was shown to increase respiration and to stimulate reactive oxygen species generation in isolated cardiac mitochondria energized with succinate. Here, we tested effects of NS1619 to alter matrix K+, H+, and swelling in mitochondria isolated from guinea pig hearts. We found that 30 μM NS1619 did not change matrix K+, H+, and swelling, but that 50 and 100 μM NS1619 caused a concentration-dependent increase in matrix K+influx (PBFI fluorescence) only when quinine was present to block K+/H+exchange (KHE); this was accompanied by increased mitochondrial matrix volume (light scattering). Matrix pH (BCECF fluorescence) was decreased slightly by 50 and 100 μM NS1619 but markedly more so when quinine was present. NS1619 (100 μM) caused a significant leak in lipid bilayers, and this was enhanced in the presence of quinine. The K+ionophore valinomycin (0.25 nM), which like NS1619 increased matrix volume and increased K+influx in the presence of quinine, caused matrix alkalinization followed by acidification when quinine was absent, and only alkalinization when quinine was present. If K+is exchanged instantly by H+through activated KHE, then matrix K+influx should stimulate H+influx through KHE and cause matrix acidification. Our results indicate that KHE is not activated immediately by NS1619-induced K+influx, that NS1619 induces matrix K+and H+influx through a nonspecific transport mechanism, and that enhancement with quinine is not due to the blocking of KHE, but to a nonspecific effect of quinine to enhance current leak by NS1619.
Regulation of circadian gene expression in the kidney by light and food cues in rats
Authors: Wu Tao , Ni Yinhua , Dong Yue , Xu Jiafeng , Song Xiaohong , Kato Hisanori , Fu Zhengwei
Published: January 2010

Although studies involving the circadian response to external time cues indicate that the peripheral clocks are dominated mainly by food cues, whether and how changes in the light and food cues affect the circadian rhythm of the renal clock is still largely unknown. In the present study, we found that the circadian phases of Bmal1, Clock, Cry1, Per1, and Per2 were altered differently by the stimuli of food and light cues in the kidney. After the individual reversal of the light-dark (LD) cycle for 7 days, Per1 displayed a 4-h phase delay, whereas the peak phases of Bmal1, Clock, Cry1 and Per2 almost remained the same as those in the control condition. With regard to the feeding-induced circadian resetting of the renal clock, we found that the resetting processes of clock genes could not be completed within 7 days, suggesting a weak synchronization effect of the food cue on the renal circadian clock. Moreover, the reentrainment of the clock genes was greatly enhanced after the reversal of both the feeding schedule and the LD cycle. Noticeably, the phases of Per1 and Clock were shifted most rapidly by 12 h within 3 days after the simultaneous reversal of the feeding schedule and the LD cycle, whereas their peak phases were only shifted by 4 h and 8 h, respectively, on the 7th day after the individual reversal of the feeding schedule. Thus Per1 and Clock may play important roles in the light-induced resetting of the circadian rhythms in the kidney.
Mechanisms of endothelium-dependent vasodilation in male and female, young and aged offspring born growth restricted
Authors: Morton J. S. , Rueda-Clausen C. F. , Davidge S. T.
Published: January 2010

Numerous epidemiological studies have shown that cardiovascular dysfunction in adult life may be programmed by compromised growth in utero. Aging is a risk factor for vascular endothelial-dependent dysfunction. After birth, the impact of intrauterine growth restriction (IUGR) on normal aging mechanisms of vascular dysfunction is not known. We hypothesized that IUGR would cause changes in vascular function that would affect the mechanisms of endothelium-dependent vasodilation later in life in an age- or sex-dependent manner. To create an IUGR model, pregnant Sprague-Dawley rats were placed in a hypoxic (12% O2) or control (room air, 21% O2) environment from days 15 to 21 of the pregnancy, and both male and female offspring were investigated at 4 or 12 mo of age. Endothelial function was assessed in small mesenteric arteries using methacholine (MCh)-induced vasodilation in a wire myograph system. The involvement of nitric oxide (NO), prostaglandins, and endothelium-derived hyperpolarizing factor (EDHF) was assessed using the inhibitors Nω-nitro-l-arginine methyl ester hydrochloride, meclofenamate, or a combination of apamin and TRAM-34 (SKCaand IKCablockers), respectively. EDHF-induced vasodilation was further investigated by using inhibitors of P450 epoxygenases [ N-methylsulfonyl-6-(2-propargyloxyphenyl) hexanamide] and gap junctions (18α-glycyrrhetinic acid). NO-mediated vasodilation was significantly reduced in aged controls and both young and aged IUGR females. EDHF-mediated vasodilation was maintained in all groups; however, an additional involvement of gap junctions was found in females exposed to hypoxia in utero, which may represent a compensatory mechanism. A change in the mechanisms of vasodilation occurring at an earlier age in IUGR offspring may predispose them to adult cardiovascular diseases.
NOX2 is the primary source of angiotensin II-induced superoxide in the macula densa
Authors: Fu Yiling , Zhang Rui , Lu Deyin , Liu Haifeng , Chandrashekar Kiran , Juncos Luis A. , Liu Ruisheng
Published: January 2010

Macula densa (MD)-mediated regulation of renal hemodynamics via tubuloglomerular feedback is regulated by interactions between factors such as superoxide (O2−) and angiotensin II (ANG II). We have reported that NaCl-induced O2−in the MD is produced by the NOX2 isoform of NADPH oxidase (NOX); however, the source of ANG II-induced O2−in MD is unknown. Thus we determined the pathways by which ANG II increased O2−in the MD by measuring O2−in ANG II-treated MMDD1 cells, a MD-like cell line. ANG II caused MMDD1 O2−levels to increase by more than twofold ( P < 0.01). This increase was blocked by losartan (AT1receptor blocker) but not PD-123319 (AT2receptor antagonist). Apocynin (a NOX inhibitor) decreased O2−by 86% ( P < 0.01), whereas oxypurinol (a xanthine oxidase inhibitor) and NS-398 (a cyclooxygenase-2 inhibitor) had no significant effect. The NOX-dependent increase in O2−was due to the NOX2 isoform; a short interfering (si)RNA against NOX2 blunted ANG II-induced increases in O2−, whereas the NOX4/siRNA did not. Finally, we found that inhibiting the Rac1 subunit of NOX blunted ANG II-induced O2−production in NOX4/siRNA-treated cells but did not further decrease it in NOX2/siRNA-treated cells. Our results indicate that ANG II stimulates O2−production in the MD primarily via AT1-dependent activation of NOX2. Rac1 is required for the full activation of NOX2. This pathway may be an important component of ANG II enhancement of tubuloglomerular feedback.
Exocyst Sec10 protects epithelial barrier integrity and enhances recovery following oxidative stress, by activation of the MAPK pathway
Authors: Park Kwon Moo , Fogelgren Ben , Zuo Xiaofeng , Kim Jinu , Chung Daniel C. , Lipschutz Joshua H.
Published: January 2010

Cell-cell contacts are essential for epithelial cell function, and disruption is associated with pathological conditions including ischemic kidney injury. We hypothesize that the exocyst, a highly-conserved eight-protein complex that targets secretory vesicles carrying membrane proteins, is involved in maintaining renal epithelial barrier integrity. Accordingly, increasing exocyst expression in renal tubule cells may protect barrier function from oxidative stress resulting from ischemia and reperfusion (I/R) injury. When cultured on plastic, Madin-Darby canine kidney (MDCK) cells overexpressing Sec10, a central exocyst component, formed domes showing increased resistance to hydrogen peroxide (H2O2). Transepithelial electric resistance (TER) of Sec10-overexpressing MDCK cells grown on Transwell filters was higher than in control MDCK cells, and the rate of TER decrease following H2O2 treatment was less in Sec10-overexpressing MDCK cells compared with control MDCK cells. After removal of H2O2, TER returned to normal more rapidly in Sec10-overexpressing compared with control MDCK cells. In collagen culture MDCK cells form cysts, and H2O2 treatment damaged Sec10-overexpressing MDCK cell cysts less than control MDCK cell cysts. The MAPK pathway has been shown to protect animals from I/R injury. Levels of active ERK, the final MAPK pathway step, were higher in Sec10-overexpressing compared with control MDCK cells. U0126 inhibited ERK activation, exacerbated the H2O2-induced decrease in TER and cyst disruption, and delayed recovery of TER following H2O2 removal. Finally, in mice with renal I/R injury, exocyst expression decreased early and returned to normal concomitant with functional recovery, suggesting that the exocyst may be involved in the recovery following I/R injury.
Genetic Variants of Nucleotide Excision Repair Genes Are Associated with DNA Damage in Coke Oven Workers
Authors: Wang Fang , He Yuefeng , Guo Huan , Li Jichao , Yang Yan , Wu Zhou , Zheng Hongyan , Wu Tangchun
Published: January 2010

We explored the effects of single nucleotide polymorphisms (SNP) of nucleotide excision repair (NER) genes on DNA damage caused by exposure to carcinogenic polycyclic aromatic hydrocarbons (PAH) in 475 Chinese workers. We quantified urinary 1-hydroxypyrene using high-performance liquid chromatography, and the DNA damage level of lymphocytes was examined by the comet assay and represented as the Olive tail moment (OTM) value. We genotyped 38 tagSNPs in 10 NER genes. The SNP function was further investigated using luciferase reporter assay in three cell lines. Our results showed that two promoter SNPs, XPA rs1800975 and XPC rs3731055, were associated with lower OTM values (Ptrend = 0.01 and 0.02 respectively). However, another missense SNP rs2228001 in the XPC gene was positively associated with OTM value (Ptrend = 0.01). A stratified analysis found that the association between this SNP and DNA damage was only observed among subjects with higher PAH exposure levels but not among those with lower exposure levels (Pinteraction = 0.018). A dose-response association was found between the combined risk alleles of the above three genetic variants and increased DNA damage levels (Ptrend = 0.004). This association was more pronounced in subjects with higher PAH exposure than those with lower exposure levels (Pinteraction = 0.046). Our functional study indicated that XPA rs1800975G and XPC rs3731055A alleles had a higher luciferase expression than their corresponding SNP alleles (P < 0.05). These results suggested that genetic variations in key NER genes, especially in XPA and XPC genes, may modulate DNA damage levels when exposed to PAHs. Cancer Epidemiol Biomakers Prev; 19(1); 211–8
Patterns of Sex Steroid Hormones in Nipple Aspirate Fluid during the Menstrual Cycle and after Menopause in Relation to Serum Concentrations
Authors: Chatterton Robert T. , Khan Seema A. , Heinz Richard , Ivancic David , Lee Oukseub
Published: January 2010

Previous studies have shown that progesterone concentrations in serum and nipple aspirate fluid (NAF) are significantly correlated in premenopausal women, but estradiol concentrations are not. We therefore sought to ascertain the patterns of both steroids in NAF throughout the menstrual cycle and in postmenopausal women. Simultaneous samples of blood and NAF were obtained from 40 premenopausal and 16 postmenopausal women. Premenopausal samples were backdated from the following menstrual period. Steroids were purified by high-performance liquid chromatography before quantification by immunoassays. Serum steroids and NAF progesterone followed the expected pattern across the menstrual cycle, with a midcycle peak of estradiol and a midluteal peak of progesterone. However, the estradiol peak in NAF occurred about a week after the serum peak in the midluteal phase, when serum estradiol had declined to less than half the value at midcycle. NAF estrone was also elevated at the midluteal phase. Potential estrogen precursors androstenedione, estrone sulfate, and dehydroepiandrosterone sulfate declined in NAF from midcycle to the midluteal phase as NAF estradiol was increasing. Progesterone concentrations were significantly lower in NAF in postmenopausal women than in premenopausal women, but estrogen concentrations were not. This is the first description of the temporal relationships of sex steroids in NAF and serum relative to the menstrual cycle. These results provide insights into the lack of correlation of NAF and breast tissue estrogens with serum estrogens, and generate new hypotheses. Cancer Epidemiol Biomarkers Prev; 19(1); 275–9
Population-Based Study of the Association of Variants in Mismatch Repair Genes with Prostate Cancer Risk and Outcomes
Authors: Langeberg Wendy J. , Kwon Erika M. , Koopmeiners Joseph S. , Ostrander Elaine A. , Stanford Janet L.
Published: January 2010

Background: Mismatch repair (MMR) gene activity may be associated with prostate cancer risk and outcomes. This study evaluated whether single nucleotide polymorphisms (SNP) in key MMR genes are related to prostate cancer outcomes.

Methods: Data from two population-based case-control studies of prostate cancer among Caucasian and African-American men residing in King County, Washington were combined for this analysis. Cases (n = 1,458) were diagnosed with prostate cancer in 1993 to 1996 or 2002 to 2005 and were identified through the Seattle-Puget Sound Surveillance Epidemiology and End Results cancer registry. Controls (n = 1,351) were age-matched to cases and were identified through random digit dialing. Logistic regression was used to assess the relationship between haplotype-tagging SNPs and prostate cancer risk and disease aggressiveness. Cox proportional hazards regression was used to assess the relationship between SNPs and prostate cancer recurrence and prostate cancer–specific death.

Results: Nineteen SNPs were evaluated in the key MMR genes: five in MLH1, 10 in MSH2, and 4 in PMS2. Among Caucasian men, one SNP in MLH1 (rs9852810) was associated with overall prostate cancer risk [odds ratio, 1.21; 95% confidence interval (95% CI), 1.02, 1.44; P = 0.03], more aggressive prostate cancer (odds ratio, 1.49; 95% CI, 1.15, 1.91; P < 0.01), and prostate cancer recurrence (hazard ratio, 1.83; 95% CI, 1.18, 2.86; P < 0.01), but not prostate cancer–specific mortality. A nonsynonymous coding SNP in MLH1, rs1799977 (I219V), was also found to be associated with more aggressive disease. These results did not remain significant after adjusting for multiple comparisons.

Conclusion: This population-based case-control study provides evidence for a possible association with a gene variant in MLH1 in relation to the risk of overall prostate cancer, more aggressive disease, and prostate cancer recurrence, which warrants replication. Cancer Epidemiol Biomarkers Prev;19(1); OF1–7
The ATM Inhibitor KU-55933 Suppresses Cell Proliferation and Induces Apoptosis by Blocking Akt In Cancer Cells with Overactivated Akt
Authors: Li Yan , Yang Da-Qing
Published: January 2010

Aberrant activation of Akt plays a pivotal role in cancer development. ATM, a protein deficient in patients with ataxia-telangiectasia disease, is traditionally considered as a nuclear protein kinase that functions as a signal transducer in response to DNA damage. It has recently been shown that ATM is also a cytoplasmic protein that mediates the full activation of Akt in response to insulin. Our study shows that a specific ATM inhibitor, KU-55933, blocks the phosphorylation of Akt induced by insulin and insulin-like growth factor I in cancer cells that exhibit abnormal Akt activity. Moreover, KU-55933 inhibits cancer cell proliferation by inducing G1 cell cycle arrest. It does so through the downregulation of the synthesis of cyclin D1, a protein known to be elevated in a variety of tumors. In addition, KU-55933 treatment during serum starvation triggers apoptosis in these cancer cells. Our results suggest that KU-55933 may be a novel chemotherapeutic agent targeting cancer resistant to traditional chemotherapy or immunotherapy due to aberrant activation of Akt. Furthermore, KU-55933 completely abrogates rapamycin-induced feedback activation of Akt. Combination of KU-55933 and rapamycin not only induces apoptosis, which is not seen in cancer cells treated only with rapamycin, but also shows better efficacy in inhibiting cancer cell proliferation than each drug alone. Therefore, combining KU-55933 with rapamycin may provide a highly effective approach for improving mammalian target of rapamycin–targeted anticancer therapy that is currently hindered by rapamycin-induced feedback activation of Akt. Mol Cancer Ther; 9(1); 113–25
Global Methylation Pattern of Genes in Androgen-Sensitive and Androgen-Independent Prostate Cancer Cells
Authors: Mishra Dhruva Kumar , Chen Zujian , Wu Yanyuan , Sarkissyan Marianna , Koeffler H. Phillip , Vadgama Jaydutt V.
Published: January 2010

Promoter DNA methylation of CpG islands is an important epigenetic mechanism in cancer development. We have characterized the promoter methylation profile of 82 genes in three prostate cancer cell lines (LNCaP, PC3, and DU145) and two normal prostate cell lines (RWPE1 and RWPE2). The methylation pattern was analyzed using a Panomics gene array system that consists of immobilized probes of known gene promoters on a nitrocellulose membrane. Methylation binding protein–purified methylated DNA was hybridized on the membrane and detected by the chemiluminescence method. We analyzed methylation profile in normal (RWPE1) versus cancerous cells and androgen receptor (AR)–sensitive (LNCaP) versus AR-negative cells (DU145 and PC3). Our study shows that >50% of the genes were hypermethylated in prostate cancer cells compared with 13% in normal cell lines. Among these were the tumor suppressor (RB, TMS1, DAPK, RBL1, PAX6, and FHIT), cell cycle (p27KIP1 and CDKN2A), transporters (MDR1, MLC1, and IGRP), and transcription factor (STAT1, CIITA, MYOD, and NPAT) genes. Relative methylation pattern shows that most of these genes were methylated from 5-fold to >10-fold compared with the normal prostate cells. In addition, promoter methylation was detected for the first time in target genes such as RIOK3, STAT5, CASP8, SRBC, GAGE1, and NPAT. A significant difference in methylation pattern was observed between AR-sensitive versus AR-negative cancer cells for the following genes: CASP8, GPC3, CD14, MGMT, IGRP, MDR1, CDKN2A, GATA3, and IFN. In summary, our study identified candidate genes that are methylated in prostate cancer. Mol Cancer Ther; 9(1); 33–45
A Human Monoclonal Anti-ANG2 Antibody Leads to Broad Antitumor Activity in Combination with VEGF Inhibitors and Chemotherapy Agents in Preclinical Models
Authors: Brown Jeffrey L. , Cao Z. Alexander , Pinzon-Ortiz Maria , Kendrew Jane , Reimer Corinne , Wen Shenghua , Zhou Joe Q. , Tabrizi Mohammad , Emery Steve , McDermott Brenda , Pablo Lourdes , McCoon Patricia , Bedian Vahe , Blakey David C.
Published: January 2010

Localized angiopoietin-2 (Ang2) expression has been shown to function as a key regulator of blood vessel remodeling and tumor angiogenesis, making it an attractive candidate for antiangiogenic therapy. A fully human monoclonal antibody (3.19.3) was developed, which may have significant pharmaceutical advantages over synthetic peptide-based approaches in terms of reduced immunogenicity and increased half-life to block Ang2 function. The 3.19.3 antibody potently binds Ang2 with an equilibrium dissociation constant of 86 pmol/L, leading to inhibition of Tie2 receptor phosphorylation in cell-based assays. In preclinical models, 3.19.3 treatment blocked blood vessel formation in Matrigel plug assays and in human tumor xenografts. In vivo studies with 3.19.3 consistently showed broad antitumor activity as a single agent across a panel of diverse subcutaneous and orthotopic xenograft models. Combination studies of 3.19.3 with cytotoxic drugs or anti–vascular endothelial growth factor agents showed significant improvements in antitumor activity over single-agent treatments alone with no apparent evidence of increased toxicity. Initial pharmacokinetic profiling studies in mice and nonhuman primates suggested that 3.19.3 has a predicted human half-life of 10 to 14 days. These studies provide preclinical data for 3.19.3 as a potential new antiangiogenic therapy as a single agent or in combination with chemotherapy or vascular endothelial growth factor inhibitors for the treatment of cancer. Mol Cancer Ther; 9(1); 145–56
BAMLET Activates a Lysosomal Cell Death Program in Cancer Cells
Authors: Rammer Paul , Groth-Pedersen Line , Kirkegaard Thomas , Daugaard Mads , Rytter Anna , Szyniarowski Piotr , Høyer-Hansen Maria , Povlsen Lou Klitgaard , Nylandsted Jesper , Larsen Jakob E. , Jäättelä Marja
Published: January 2010

A complex of human α-lactalbumin and oleic acid (HAMLET) was originally isolated from human milk as a potent anticancer agent. It kills a wide range of transformed cells of various origins while leaving nontransformed healthy cells largely unaffected both in vitro and in vivo. Importantly, purified α-lactalbumins from other mammals form complexes with oleic acid that show biological activities similar to that of HAMLET. The mechanism by which these protein-lipid complexes kill tumor cells is, however, largely unknown. Here, we show that complex of bovine α-lactalbumin and oleic acid (BAMLET), the bovine counterpart of HAMLET, kills tumor cells via a mechanism involving lysosomal membrane permeabilization. BAMLET shows potent cytotoxic activity against eight cancer cell lines tested, whereas nontransformed NIH-3T3 murine embryonic fibroblasts are relatively resistant. BAMLET accumulates rapidly and specifically in the endolysosomal compartment of tumor cells and induces an early leakage of lysosomal cathepsins into the cytosol followed by the activation of the proapoptotic protein Bax. Ectopic expression of three proteins known to stabilize the lysosomal compartment, i.e. heat shock protein 70 (Hsp70), Hsp70-2, and lens epithelium-derived growth factor, confer significant protection against BAMLET-induced cell death, whereas the antiapoptotic protein Bcl-2, caspase inhibition, and autophagy inhibition fail to do so. These data indicate that BAMLET triggers lysosomal cell death pathway in cancer cells, thereby clarifying the ability of α-lactalbumin:oleate complexes to kill highly apoptosis-resistant tumor cells. Mol Cancer Ther; 9(1); 24–32
The Preclinical Pharmacology and Therapeutic Activity of the Novel CHK1 Inhibitor SAR-020106
Authors: Walton Michael I. , Eve Paul D. , Hayes Angela , Valenti Melanie , De Haven Brandon Alexis , Box Gary , Boxall Kathy J. , Aherne G. Wynne , Eccles Suzanne A. , Raynaud Florence I. , Williams David H. , Reader John C. , Collins Ian , Garrett Michelle D.
Published: January 2010

Genotoxic antitumor agents continue to be the mainstay of current cancer chemotherapy. These drugs cause DNA damage and activate numerous cell cycle checkpoints facilitating DNA repair and the maintenance of genomic integrity. Most human tumors lack functional p53 and consequently have compromised G1-S checkpoint control. This has led to the hypothesis that S and G2-M checkpoint abrogation may selectively enhance genotoxic cell killing in a p53-deficient background, as normal cells would be rescued at the G1-S checkpoint. CHK1 is a serine/threonine kinase associated with DNA damage–linked S and G2-M checkpoint control. SAR-020106 is an ATP-competitive, potent, and selective CHK1 inhibitor with an IC50 of 13.3 nmol/L on the isolated human enzyme. This compound abrogates an etoposide-induced G2 arrest with an IC50 of 55 nmol/L in HT29 cells, and significantly enhances the cell killing of gemcitabine and SN38 by 3.0- to 29-fold in several colon tumor lines in vitro and in a p53-dependent fashion. Biomarker studies have shown that SAR-020106 inhibits cytotoxic drug–induced autophosphorylation of CHK1 at S296 and blocks the phosphorylation of CDK1 at Y15 in a dose-dependent fashion both in vitro and in vivo. Cytotoxic drug combinations were associated with increased γH2AX and poly ADP ribose polymerase cleavage consistent with the SAR-020106–enhanced DNA damage and tumor cell death. Irinotecan and gemcitabine antitumor activity was enhanced by SAR-020106 in vivo with minimal toxicity. SAR-020106 represents a novel class of CHK1 inhibitors that can enhance antitumor activity with selected anticancer drugs in vivo and may therefore have clinical utility. Mol Cancer Ther; 9(1); 89–100
Structure Activity Analysis of 2-Methoxyestradiol Analogues Reveals Targeting of Microtubules as the Major Mechanism of Antiproliferative and Proapoptotic Activity
Authors: Chua Yee Shin , Chua Yee Liu , Hagen Thilo
Published: January 2010

2-Methoxyestradiol (2ME2) is an anticancer agent with antiproliferative, antiangiogenic, and proapoptotic effects. A major proposed mechanism of drug action is the disruption of the microtubule skeleton, leading to the induction of cell cycle arrest and apoptosis. In addition, other mechanisms of action have been proposed, including the generation of reactive oxygen species (ROS), inhibition of hypoxia-inducible factor (HIF), and interference with mitochondrial function. In this study, we used a selection of 2ME2 analogues to conduct structure activity analysis and correlated the antiproliferative and proapoptotic activity of the various analogues with their effects on different drug targets. A good correlation was observed between drug activity and effects on microtubule function. In contrast, our results indicate that effects on ROS, HIF, and mitochondria are unlikely to contribute significantly to the cellular activity of 2ME2. Thus, our data indicate that the structural requirements for inducing ROS and inhibition of complex I of the mitochondrial electron transport chain were different from those required for proapoptotic drug activity. Furthermore, antioxidant treatment or overexpression of catalase did not inhibit the cellular activity of 2ME2 in epithelial cancer cells. Inhibition of HIF required much higher concentrations of 2ME2 analogues compared with concentrations that inhibited cell proliferation and induced apoptosis. Our results thus provide a better insight into the mechanism of action of 2ME2 and reveal structural requirements that confer high cellular activity, which may aid future drug development. Mol Cancer Ther; 9(1); 224–35
Cross-Species Chromosome Painting Corroborates Microchromosome Fusion during Karyotype Evolution of Birds
Authors: Hansmann T. , Nanda I. , Volobouev V. , Yang F. , Schartl M. , Haaf T. , Schmid M.
Published: January 2010

The stone curlew, also known as thick-knee (Burhinus oedicnemus, BOE), represents a phylogenetically young species of the shorebirds (Charadriiformes) that exhibits one of the most atypical genome organizations known within the class of Aves, due to an extremely low diploid number (2n = 42) and only 6 pairs of microchromosomes in its complement. This distinct deviation from the ‘typical’ avian karyotype is attributed to repeated fusions of ancestral microchromosomes. In order to compare different species with this atypical avian karyotype and to investigate the chromosome rearrangement patterns, chromosome-specific painting probes representing the whole genome of the stone curlew were used to delineate chromosome homology between BOE and 5 species belonging to 5 different avian orders: herring gull (Charadriiformes), cockatiel (Psittaciformes), rock pigeon (Columbiformes), great gray owl (Strigiformes) and Eurasian coot (Gruiformes). Paints derived from the 20 BOE autosomes delimited 28 to 33 evolutionarily conserved segments in the karyotypes of the 5 species, similar to the number recognized by BOE paints in such a basal lineage as the chicken (28 conserved segments). This suggests a high degree of conservation in genome organization in birds. BOE paints also revealed some species-specific rearrangements. In particular, chromosomes BOE1–4 and 14, as well as to a large extent BOE5 and 6, showed conserved synteny with macrochromosomes, whereas homologous regions for BOE7–13 are found to be largely distributed on microchromosomes in the species investigated. Interestingly, the 6 pairs of BOE microchromosomes 15–20 appear to have undergone very few rearrangements in the 5 lineages investigated. Although the arrangements of BOE homologous segments on some chromosomes can be explained by complex fusions and inversions, the occurrence of homologous regions at multiple sites may point to fission of ancestral chromosomes in the karyotypes of the species investigated. However, the present results demonstrate that the ancestral microchromosomes most likely experienced fusion in the stone curlew lineage forming the medium-sized BOE chromosomes, while they have been conserved as microchromosomes in the other neoavian lineages.
Dynamic Epidemiology of Acute Viral Hepatitis in Japan
Authors: Yano Koji , Tamada Yoko , Yatsuhashi Hiroshi , Komori Atsumasa , Abiru Seigo , Ito Kiyoaki , Masaki Naohiko , Mizokami Masashi , Ishibashi Hiromi
Published: January 2010

The epidemiology of acute viral hepatitis (AVH) is dynamic and affected by many factors including hygiene, socioeconomic status and vaccination coverage. A total of 4,302 cases of AVH were sequentially enrolled in this nationwide study between 1980 and 2008. Of the cases of AVH, acute hepatitis A (AHA) accounted for 1,583 (36.8%), acute hepatitis B (AHB) for 1,197 (27.8%), acute hepatitis C (AHC) for 359 (8.3%), and non-A, non-B and non-C (non-ABC) for 1,163 (27.0%). Between 1980 and 1995, the proportions of AHA, AHB, AHC and non-ABC were approximately 40, 25, 10 and 25%; between 1996 and 2003, they were approximately 30, 30, 10 and 30%, and this shifted to approximately 10, 40, 10 and 40% in the last 5 years. The number of AHB caused by genotype A, which is not indigenous to Japan, was 6.0% between 1991 and 1996 but has been markedly increasing since 2000, to reach 52% in 2008. Autochthonous acute hepatitis E (AHE) accounted for 10–15% of non-ABC hepatitis after 2002. The etiology of AVH in Japan has been drastically changing. A marked increase of AHB genotype A and constant occurrence of autochthonous AHE require attention, and necessary measures should be taken.
Paradoxically Increased Ferritin Level in a Beta-Thalassemia Major Patient following the Start of Deferasirox Chelation Therapy
Authors: Ricchi Paolo , Ammirabile Massimiliano , Spasiano Anna , Costantini Silvia , Cinque Patrizia , Di Matola Tiziana , Prossomariti Luciano
Published: January 2010

In this report we present a 37-year-old thalassemia patient with hyperferritinemia referred to our Microcytemia Center at the beginning of deferasirox (DFX) therapy. Treatment with subcutaneous infusions of desferrioxamine (DFO) had started when he was 10 years old. During the 6-month DFX treatment, serum ferritin levels progressively increased from 600 to 2,700 ng/ml despite progressive DFX dose adjustments. This paradoxically abnormal ferritin levels required drug discontinuation but were not paralleled by a similar iron burden in T2* magnetic resonance imaging. In this clinical case, ferritin levels were inappropriately increased following initiation of DFX treatment, but in the presence of an almost unmodified pattern of organ iron overload. Excluding the diagnostic dilemma of an improbable failure of DFX chelation, the pathogenesis of this phenomenon remains to be clarified, thus further complicating the problem of ferritin specificity and its role in monitoring chelation efficacy and in adapting DFX dosage in a limited period of treatment.
Influence of Cochlear Function on Auditory Temporal Resolution in Tinnitus Patients
Authors: Sanches Seisse Gabriela Gandolfi , Sanchez Tanit Ganz , Carvallo Renata Mota Mamede
Published: January 2010

Our aim was to analyze the influence of subtle cochlear damage on temporal auditory resolution in tinnitus patients. Forty-eight subjects (hearing threshold ≤25 dB HL) were assigned to one of two experimental groups: 28 without auditory complaints (mean age, 28.8 years) and 20 with tinnitus (mean age, 33.5 years). We analyzed distortion product otoacoustic emission growth functions (by threshold, slope, and estimated amplitude), extended high-frequency thresholds, and the Gaps-in-Noise test. There were differences between the groups, principally in the extended high-frequency thresholds and the Gaps-in-Noise test results. Our findings suggest that subtle peripheral hearing impairment affects temporal resolution in tinnitus, even when pure-tone thresholds as conventionally measured appear normal.
Growing Teratoma Syndrome of the Ovary after Fertility-Sparing Surgery and Successful Pregnancy
Authors: Matsushita Hiroshi , Arai Kei , Fukase Masayuki , Takayanagi Takeshi , Ikarashi Hirokazu
Published: January 2010

Background: Growing teratoma syndrome (GTS) is rare and is defined as an enlarging mature teratoma that arises during or after chemotherapy for a malignant germ cell tumor, with normalization of previously elevated serum tumor markers. Case: A 30-year-old nulliparous Japanese woman was diagnosed as having a stage IIIa immature teratoma. After fertility-sparing surgery, she received 4 cycles of chemotherapy consisting of cisplatin, etoposide and bleomycin. Thereafter, she successfully gave birth twice. Eight years after the initial surgery, a mass mimicking an adrenal tumor was resected laparoscopically. Surgical specimens revealed a mature teratoma, and she was diagnosed as having GTS postoperatively. Conclusion: Clinicians should consider that GTS may present late, even after pregnancy. Therefore, long-term follow-up of patients treated for ovarian immature teratoma should be mandatory.
Bacterial and Eukaryotic Phosphoketolases: Phylogeny, Distribution and Evolution
Authors: Sánchez Borja , Zúñiga Manuel , González-Candelas Fernando , de los Reyes-Gavilán Clara G. , Margolles Abelardo
Published: January 2010

Phosphoketolases (XFPs) are glycolytic enzymes present in several organisms belonging to the Eukarya and Bacteria domains. A total of 151 putative xfp genes were detected in 650 complete genomes available in public databases. Elimination of redundant sequences and pseudogenes rendered a final data set of 128 phosphoketolases, which was analyzed by phylogenetic methods. The distribution of xfp genes was uneven in most taxonomic groups, with the exception of the taxonomical division Lactobacillaceae, in which all the species studied harbored a putative xfp gene. Putative xfp genes were also present predominantly in Rhizobiales and Actinobacteria divisions, in which 23 out of 28 genomes and 23 out of 41 genomes contained at least one putative xfp homolog, respectively. Phylogenetic analyses showed clear discordance with the expected order of organismal descent even in groups where xfp is prevalent, such as Lactobacillaceae. The presence of putative paralogs in some organisms cannot account for these discrepancies; instead, these paralogs are most possibly xenologs. The results of the phylogenetic analyses, the distribution of xfp genes and the location of some xfp genes in plasmids are independent pieces of evidence that point to horizontal gene transfer as a major driving force in the evolution of phosphoketolases.
Changes in the Tracheal Respiratory Mucosa of Rats Following Experimentally Induced Burns of the Skin
Authors: Femenić B.
Published: January 2010

The tracheal respiratory mucosa of rats sacrificed on the second and the fourth day following experimentally induced burns of the skin was submitted to histological study. Pathological changes were found in the respiratory mucosa of all animals with hepatic and renal damage.
DC-Determined Displacement of the Nystagmus Beat in Rotatory Tests
Authors: Lundgren A. , Tibbling L. , Henriksson N.G.
Published: January 2010

The deviation of the eyes induced by vestibular stimuli is defined as the distance in degrees from the mid-position of the eyes to the position to which the eyes return in the fast phase. At an intensive angular stimulus the eyes are found to lead the head and to such an extent that the whole nystagmus beat can be displaced in the direction of the fast phase. The deviation of the eyes in the fast direction seems to be closely related to the deflection of the cupula and subsides exponentially in the postacceleratory period in about the same way as does the speed of the slow component. The eyes, however, reach their mid-position before the nystagmus disappears, indicating that a certain degree of intensity in the stimulus is needed to make the eyes deviate in the direction of the fast phase.
La pathogénie d’action et l’effet thérapeutique de la cortisone sur les oesophagites corrosives aiguës
Authors: Taillens J.-P.
Published: January 2010

The author first reviews the action of corticoids in general, then their beneficial as well as unwanted effects on a corrosive lesions of the oesophagus. Management depends above all on the pathology of the lesions, in practice on endoscopic findings. Local contra-indications are severe necrosis and profuse gastric bleeding. Distal (i.e. gastric) injury is more frequent than is commonly realised as ingestion of corrosives precludes its investigation. The experience of 45 cases has shown that 50% of acid and 70% of lye burns will benefit from corticolherapy. If prescribed, it must be given from the third day onwards; earlier administration will impair the proliferation of fibroblasts necessary for repair; afterwards it will be too late to forestall their transformation into fibrocytes responsible for the future and already unpreventable stenosis.
Results of and Experience with Audiometric Examination of Normally Hearing Children
Authors: Lesak J.
Published: January 2010

The introduction deals with the description of some achievements of the advanced Danish audiology, and the equipment corresponding to this, in the paedoaudiometric department in the State Hearing Centre in Odense. The author has examined groups of normally hearing children aged from 4 months to 12 years at this Centre. They were outpatient and mostly single examinations which is the most frequent routine method used at the Centre. The audiometric part deals with the results of examination with headphones, in the free field, using pure tones and with special filtred programmes, and of threshold values with and without sound localization. The situation, reaction, interest, cooperation and sound localizing power of the child during examination, and the results at the play-board and with a series of other equipment are the subject of the psychological part. The aim of the investigation was to obtain basic, normal data and information for the Centre to permit the assessment of audiograms in outpatient paedoaudiometric practice as normal or pathological. The results may perhaps be interesting and useful for purposes of comparison for other paedoaudiometric instituts.
Les adénopathies malignes épithéliales en apparence primitives du cou
Authors: Nicoucar G.R. , Gainer H.
Published: January 2010

The diagnosis of primary malignant epithelial cervical nodes is applied to patients having epithelial metastatic ganglions where, despite complete and careful examinations of all systems, it was not possible to discover the primary cancerous focus. Many clinicians think that these tumors arise from epithelial degeneration of bronchial embryonic remnants in the latero-cervical region and group, without any anatomo-pathological proof, the total of these formations under the title of malignant branchioma. While some remnants may exceptionally degenerate, it still remains that the great majority of what one calls malignant branchioma, are nothing more than metastatic ganglions of the neck. This conception based on anatomo-pathological research is so close to reality that some practioners even doubt the existence of malignant branchiomas. During the last 15 years (1954–1969) 558 patients suffering from carcinoma of the oto-rhino-laryngological sphere have been hospitalized and regularly followed by the Geneva oto-rhino-laryngological clinic and polyclinic. Of this important material which we have studied with care, 12 cases (2.1%) of malignant cervical epithelial adenopathies, seemingly primary, were observed. The anatomopathological examination of these cervical tumors revealed in each case the picture of a metastatic invasion of the ganglion. In no case were we able to put in evidence kysts, or embryonic inclusions with or without degeneration. On the other hand, in studying the anatomo-pathological report of branchiogenic or thyreoglossal kysts operated in our clinic during the past 10 years, we found no trace of malignant transformation. Among our 12 patients, 3 (25%) died 8–12 months after the appearance of the first symptoms without having received any treatment. At the autopsy of 2 of them the primary tumor was found once at the level of the cavum, and once at the level of the parotid. The 3 other patients (25%) who received a partial treatment (removal of ganglions or cobalt therapy) died 6–16 months later, and at the autopsy we discovered in 2 cases the primary tumor (tonsil and parotid) while in the 3rd case all research remained in vain. Of the 6 other patients who received a complete treatment (surgery and radiotherapy over large areas), 4 (33%) are alive and present no further symptoms, 2 (17%) died, 1–3½ years after the first symptoms. The primary tumors in the 2 latter patients were found in the thyroid gland and in the bronches. The appearance of the primary tumor during the lifetime of the patienfs as well after the end of the treatment was observed in 2 of our patients. In 1 case it was found at the level of the piriform sinus 1½ years after the first symptoms, and in the other case at the level of the bronchi 3 years after their appearance.
Compressions œso-trachéales par malformations artérielles
Authors: Rivier A.
Published: January 2010

After having remembered different types of vascular and arterial anomalies provoking esotracheal compressions, the author presents three cases recently examined at the Ear, Nose and Throat Clinic in Lausanne (Prof. J.-P. TAILLENS): a right aortic arch with esotracheal compression in a 9-year-old child, an aberrant right sub-clavian artery (provoking disphagia lusoria), and a right aortic arch with aneurism of the aortic cross and left arterial ligament in a 78-year-old patient with sub-total obstruction of the trachea by the vascular anomalies.
The Function of the Interdental Cells of the Limbus Spiralis
Authors: Mira E.
Published: January 2010

The pattern of several oxidative and hydrolytic enzyme activities has been studied in the interdental cells of the limbus spiralis of the cochlea of the guinea pig, using histochemical methods. Some of the interdental cells show a high activity for NADH-diaphorase, isocitric dehydrogenase, succinic dehydrogenase and acid phosphatase. A marked difference of intensity of reaction among the cells is present, suggesting a different metabolic rate. The functional meaning of the findings, and the possible participation of the interdental cells in the processes of secretion and reabsorption of endolymph is discussed.