In summary, in this study of more than 40,000 LAIV recipients 5–17 years of age, rates of MAEs and SAEs were compared between LAIV-vaccinated individuals and multiple nonrandomized controls. SAEs and hospitalizations after vaccination with LAIV were uncommon, and no pattern of MAEs was found to occur at higher rates than control groups. The results of this study are consistent with preapproval Volasertib mouse studies [3], [13] and [14] and with reports to the Vaccine Adverse Events Reporting System in the years after the initial approval of

LAIV [12], which demonstrated no significant adverse outcomes after receipt of LAIV by eligible individuals 5–17 years of age. A similar study is currently underway in children 2–4 years of age. Contributors: Study concept and design: Drs. Baxter, Toback, Sifakis, and Ambrose, Mr. Hansen, Ms. Bartlett, Ms. Aukes, and Mr. Lewis. Acquisition of data: Dr. Baxter, Mr. Hansen, Ms. Bartlett, Ms. Aukes, and Mr. Lewis. Analysis and interpretation of data: all authors. selleckchem Drafting of the manuscript:

all authors. Critical revision of the manuscript for important intellectual content: all authors. Statistical analysis: Ms. Bartlett and Dr. Wu. All authors have seen and approved the final manuscript for submission. Financial disclosures: Drs. Toback, Sifakis, Wu, and Ambrose are employees of MedImmune, LLC, Gaithersburg, MD. Dr inhibitors Baxter receives grants from Merck, GSK, Novartis, and Sanofi Pasteur. Funding/support: This research was funded by MedImmune. Role of the sponsor: Employees of MedImmune worked collaboratively with the investigators in the design of the study, in analysis Vasopressin Receptor and interpretation

of the data, and reviewed and approved the manuscript. Additional contributions: Editorial assistance in formatting the manuscript for submission was provided by Susan E. Myers, MSc, and Gerard P. Johnson, PhD, of Complete Healthcare Communications, Inc. (Chadds Ford, PA) and funded by MedImmune. “
“T cells are important mediators of the adaptive immune response against infections caused by intracellular microorganisms, including the digenetic intracellular protozoan parasite Trypanosoma cruzi, the causative agent of Chagas disease (American trypanosomiasis). Genetic deficiency or specific treatments leading to the depletion of CD4+ or CD8+ T cells critically impairs the acquired immunity observed during experimental mouse infection [1], [2], [3] and [4]. Although, the anti-parasitic effect exerted by the T cells is largely mediated by IFN-γ, other mediators may also participate in the efficient elimination of parasites from the host [1], [2], [3] and [4]. In inbred mouse strains or humans, MHC class II-restricted CD4+ T cells recognize multiple antigens from T. cruzi [5], [6], [7], [8] and [9], whereas MHC class Ia-restricted CD8+ T cells are primarily specific for immunodominant epitopes that are expressed by surface antigens members of a large family of T.

Most high-income see more countries in Asia are affected by non-communicable diseases. However, the prevalence of CVD risk factors is still lower compared to the USA, Europe and the world, except for smoking. Within Asia, men in high-income countries tend to smoke less compared to middle- and low-income countries but they drink more alcohol. Lower alcohol consumption in Asia is probably contributed by alcohol abstinence in Islamic countries. Higher-income countries often have higher prevalence of high total cholesterol and obesity, and this is contributed by their sedentary lifestyle and dietary factor (Tong et al., 2011). The drop in the mean systolic blood pressure in high-income countries might

be contributed by wider anti-hypertensive drugs used, which may not be readily available in the lower-income countries (Danaei et al., 2011). Comparing to lower-income nations, people in high-income MLN0128 cost countries tend consume more added sugars and fats, which subsequently lead to higher mean

BMI for high-income countries (Drewnowski, 2003). This study has a few limitations. Although we extracted data from the WHO database, the quality of data reported by individual country may vary. Some of the data might not be updated and there is a limit to trend data. Summarizing the prevalence of risk factors in Asia by using a simple average might not accurately reflect the distribution of data across Asia. In addition, the use of arbitrary criteria for BMI ≥ 25 kg/m2 (Asia: ≥ 23 kg/m2) may not be appropriate for the Asian population. This is the first study that systematically documents the status of men’s health in Asia which confirms

that Asian men have a shorter life expectancy and higher mortality compared to Asian women. These findings are consistent with those found in the rest of the world. We found that in Asia, men in the middle-income countries are facing a double disease crisis and there is a rising trend in cardiovascular risk factors. This imposes a significant healthcare burden which calls for a concerted effort to find solutions to address men’s health issues in Asia. The authors declare Cell press that there is no conflict of interest. The authors confirmed that there is no funding received in this study. “
“The authors regret that there is an error of consistency Libraries between what is in the Abstract and text (both correct) and the printing of Table 2 and Table 3 and Fig. 2 (all three are incorrect) for the above-referenced article. The incorrect items are from a previous version and contain 18 instead of the correct 22 samples analyzed. The interpretation and conclusion of the meta-analysis are unaffected. The authors apologize for these errors. The corrected tables and figure appear here: Table 2. Coding information for studies (K = 22) meeting inclusion criteria. “
“Due to a typesetting error, Table 1 in the above-referenced article was a copy of Table 3, rather than the real Table 1.

10 Aaptamines is marine alkaloid which has a unique structure 1H-benzo [d,e][1,6]naphthyridine and an important role since its capability to block CDK-Cyclin Complex activity. CDK-Cyclin Complex itself is an important protein complex HIF-1�� pathway which influences on abnormal cell proliferation or cancer initiator. The new aaptamine compounds i.e. aaptamines, bisdemethylaaptamine6,

and bisdemethylaaptamine-9-O-sulfate 7 and 8,9-dimethoxy-4-methyl-4H-benzo[de] [1,6snaphthyridine (2,4-methylaaptamine) was also reported able to have antiviral activity against herpes simplex type 1 virus and anti cell line. 12, 13 and 16 Sponges are among the most promising groups, and compounds with cytotoxic and antitumor activity are the most frequently found in these organisms.9 Isolation of the alkaloid 4-methylaaptamine from the marine sponge

Aaptos sp (collected in Abrolhos, Bahia, Brazil) and the preliminary activity of its crude extract to inhibit 76% of HSV-1 replication in Vero cells at a concentration of 2.4 μg/mL was first reported by Coutinho et al. 11 Many polyacethylenic macrolide compounds of marine sponges indicate cytotoxic activity; while other metabolites have antifungal activity. There had also been some reports on the secondary metabolites that could be isolated from various species of sponges in Indonesia. 14 The compounds included alcaloidhalicyclamine A – a macrolide isolated from Haliclona sp, cytotoxic alcaloid 8 – hydrosimanzamine A isolated from pachypellina sp. Bestadin-derived compounds (bastadin 16 and 17) of Lanthella basta were isolated from Sulawesi. Indonesia http://www.selleckchem.com/products/Romidepsin-FK228.html is a maritime country with substantial potentials

of marine organisms that are not yet fully utilized as the sources of bioactive substances. The studies almost conducted with marine natural products during the last decades had uncovered many substances with biomedical potential, which raised the interest of many research groups towards these ecosystems as a source of new drugs. 15 Finally, we bring to the attention that this is the first scientific report of any nature on species collected from Pecaron Bay Pasir Putih Situbondo, Jawa Timur. Few studies had been conducted at this site and very little is known about the local fauna, especially when concerning the invertebrates populations. This study is part of a more comprehensive project, which Modulators focuses on the pharmacological potential of the yet poorly explored Pecaron Bay Pasir Putih Situbondo. Further steps for this work have already been taken and deeper studies on chemical and pharmacological aspects of the most interesting species are already in progress. The rich diversity in bioactive compounds from sponges has provided molecules that interfere with the pathogenesis of a disease at many different points, which increase the chance of developing selective drugs against specific targets.

11 Guidelines advise to not lift heavy weights or children and to avoid doing repeated activities.2 and 20 Recent studies, however, have reported that weight training did not induce or exacerbate BCRL when it was performed under supervision with slow progression.21 and 22 This type of exercise results in robust functional, physiological, psychological CX5461 and clinical benefits.4 Progressive

weight training is intended to elicit benefits in health and performance by challenging skeletal muscles with controlled physiological stress to the onset of muscle fatigue. These weight-training sessions are followed by an optimal interval of rest, ranging from 48 to 72 hours; this allows physiological adaptation to occur.23 and 24 Aside from local effects at the arm, weight training has many other benefits, including: a reduction in cancer-related fatigue,25 and improvement in body weight, psychological well being,26 bone density,27 body image28 and survival.29 Some narrative19

and systematic4, 11, 18, 30 and 31 inhibitors reviews have been published on this topic. However, these reviews included studies with mixed exercise interventions30 or included non-randomised studies.4 and 18 Furthermore, at least two more randomised trials have been published since these previous reviews.4, 18 and 31 Therefore, this present review was considered to be necessary and sought to answer these research questions: 1. Is weight-training exercise safe for women with or at risk of lymphoedema after breast cancer? The following databases were searched electronically Pomalidomide purchase from inception to July/August 2012: PubMed, EMBASE, PsycINFO, CINAHL, AMED, Cochrane, PEDro, SPORTDiscus and Web of Science. Date restriction, female gender limit and peer review were applied to the results where possible. In addition, reference lists

of the identified studies GBA3 and previous reviews were searched for any potential articles. Furthermore, distinguished authors from this research area were contacted through email for any missed and relevant studies. Three key terms, ‘weight training’, ‘lymphoedema’ and ‘breast neoplasm’, were used to generate an exhaustive list of key words. Appendix 1 (see eAddenda) shows the full search strategies. Eligibility assessment of each study was conducted in a non-blinded and standardised manner by a single researcher (VP) under the supervision of the second author (DR) in three stages and every effort was undertaken to avoid subjective bias.32 In the first stage, articles obtained through the database searches were compared for duplicate entries using the de-duplicating facility of reference management softwarea and were manually cross checked. The titles and abstracts of the remaining articles were examined for eligibility against the pre-defined criteria, as presented in Box 1. Articles that were not definitely excluded by this screening were obtained in full text for further assessment.

Respondents with missing information on any variable described above are excluded.

Logistic regression in Stata 12 SE is used, and coefficients are average marginal effects (AME) predicted with the margins option. Contrary BKM120 to what is often believed, log-odds ratios or odds ratios are not comparable across studies or models ( Mood, 2010 and Wooldridge, 2002). Therefore, AME are reported, which are easily interpretable as the average impact on the probability (0–1) of good health. For categorical variables, AME give the discrete difference in the probability of good health between the relevant category and the reference group. As the outcome is restricted to be 0 or 1 the estimated effects are not additive: If a person has many risk factors, the measured outcome can still not be worse than “not good.” FGFR inhibitor The predicted probabilities of

good health in 2000 at different combinations of risk factors will therefore also be shown, using a type case, and varying the statistically significant lifestyle factors one by one and in combination for this case. The type case is a woman of average age, income and education, who usually drinks less than two glasses, eats vegetables daily, is not overweight, and does not see friends and family often (smoking, exercise and social support are set to vary). Because of Modulators sample size restrictions, response categories for some variables have been collapsed. In these cases, different categorizations have been tested, and those reported give the most robust results. Descriptives for all variables are given in Table 1. Recall that all respondents had good health in 1991, so the 20% reporting less than good self-rated health in 2000 or 2010 have seen deterioration. There are equal shares of men and women, and the average age in 1991 is 38 for respondents observed in 2000 and 36 for those observed in 2010 (this decline is explained by panel ageing, as those who remained in 2010 were younger in 1991 than those who remained in 2000). Around 30% are single households, and 28% are overweight in 1991. A majority, 74%, exercise each week, and around 60%

eat vegetables every day. 49% have never smoked, and around 30% currently smoke. Less than 10% never STK38 drink alcohol, and of those who drink, around half usually drink more than a couple of glasses. Around half the sample see friends often and an equal share see family often. Only 4% lack social support. Table 2 gives regression results for self-rated health in 2000 (models 1A–1B) and in 2010 (models 2A–2B). In both cases, model A includes lifestyle variables, and model B additionally includes control variables. Model 1A shows that weekly exercise, usually drinking more than two drinks, and seeing friends often in 1991 are positively related to health in 2000 (statistically significant, P < 0.05), while smoking and lack of social support are negatively related to health (P < 0.05).

Amphoterecin-B and Ketoconazole were used as the reference antifungal agent. The result revealed that most of newly synthesised 3,4,5-triarylisoxazole compounds exhibited good antifungal Modulators activities against F. oxysporus and C. albicans. We synthesised a series of Novel 3,4,5-triarylisoxazoles derivatives in high yields. The advantages are the usage of low cost starting BIBW2992 order chemicals and simple experimental

procedure. These derivatives are having good antifungal activity. All authors have none to declare. The authors express their thanks to Islamiah College, Vaniyambadi for the laboratory facilities provided to carry out the research work. “
“La dystrophie myotonique de type 1 est la myopathie la plus fréquente chez l’adulte. Le risque de développer une tumeur est plus élevé chez les patients atteints de dystrophie myotonique que dans la population générale. “
“Although most pharmacognostic studies focus on plants, other types of organisms are also regarded as pharmacognostically interesting. Euglena gracilis is a microalgae member of the Euglenoids,

that can grow autotrophically, heterotrophically or Hormones antagonist myxotrophically that it has been extensively studied, 1 and 2 mainly on primary metabolites production, 3, 4 and 5 but little is known about secondary metabolites biosynthesis. The most startling findings about this species concern to 4α-methylsterols, detected in trace amounts. 6 and 7E. gracilis has a wide range of nutritional requirements, suggesting the existence mafosfamide of diverse physiological patterns, generating different metabolites and/or variation in the proportion they are biosynthesised. The aim of this work is to carry out a preliminary study on two strains of E. gracilis cultured in vitro,

both in their photosynthetic and bleached forms, on their exponential and stationary growth phase. The Euglena reserve polysaccharide paramylon has been previously shown to have general antitumoral properties and reduce the negative effects of stressors. 8 and 9 Since paramylon precipitates in ethanol, our work explores the antioxidant and antitumoral in vitro effect of the extracts in its absence. Two E. gracilis strains were used: a commercial (UTEX-753) and a wild type strain (MAT) isolated from Matanza River. 10 Studies were performed on the photosynthetic (ph) strains and their bleached (b) counterparts, obtained by treatment with streptomycin. The cultures were grown in a growth chamber at 24 ± 1 °C, with 12:12 cool-white fluorescent light (150 μE m−2 s−1 irradiance) in EGM medium. 11 Cells were quantified with Neubauer’s chambers and biomass was obtained via centrifugation at 4 °C after 72 h (exponential phase, -EX) and 144 h of growth (stationary phase, -ST). Biomass was washed four times with distilled water at 4 °C, and then dried by lyophilisation. A general extraction was performed in all dried samples obtained with ethanol 96° and fractionated by pH changes, and partitioned with different polarity solvents (Fig.

The use of a destabilized form of CreERT2, the

inclusion of endogenous sequences in the Fos and Arc 3′UTRs that contribute to mRNA destabilization, the development of new CreER ligands that are more rapidly absorbed and metabolized than 4-OHT, and the development of drug-dependent recombinases with reduced leakiness and improved inducibility may result in an improved signal-to-noise ratio. Nonetheless, complex experiences, such as exploration of a novel environment, can increase TRAPing above homecage levels ( Figure 6), suggesting that the current version of TRAP has sufficient signal-to-noise ratio FK228 without sensory deprivation. Despite the limitations, we have shown that TRAP provides valuable genetic access to active populations of neurons with feature selectivity in multiple systems. Thus, TRAP can be used in combination with various Cre-dependent effectors to trace connectivity, record activity, and manipulate functions of these select neuronal populations. Although previous

methods, such as TetTag, also enabled genetic manipulation of functionally defined neuronal populations, TRAP’s superior temporal SAHA HDAC nmr resolution and its ability to provide permanent genetic access make it a major advancement that has the potential to enable previously impossible experiments. Methods for mouse production and histology can be found in the Supplemental Experimental Procedures. All mouse procedures were approved by the Stanford University Administrative Panel on Laboratory Animal Care and were in accordance with all applicable regulatory standards. ArcCreER (JAX stock #021881) and FosCreER (JAX stock #021882) mice can be requested from the Jackson Laboratory. In pilot experiments,

we tested a range of TM doses (30–150 mg/kg) and found that TM-induced recombination was highly nonlinear. Low TM doses (30 mg/kg TM) induced minimal recombination, particularly in the less-sensitive FosTRAP mice (data not shown). Given that 150 mg/kg TM induced robust recombination and was Rolziracetam well tolerated by the mice, this dose was used for further studies. Similarly, 15 mg/kg 4-OHT induced minimal recombination in FosTRAP mice, whereas 150 mg/kg 4-OHT was not well tolerated. For additional studies, 50 mg/kg was used. In V1 (see Figure 4), treatment with 150 mg/kg TM and 50 mg/kg 4-OHT produced similar total numbers of TRAPed cells both in the dark condition (4-OHT, 622 ± 110 cells/mm3; TM, 777 ± 191 cells/mm3; t[7] = 0.65, p = 0.53) and when administered at the time point for optimal TRAPing (0 hr 4-OHT, 5184 ± 605 cells/mm3; 24 hr TM, 5736 ± 731 cells/mm3; t[9] = 0.57, p = 0.59). We also found that 4-OHT produced more consistent results than TM. In 5%–20% of mice treated with TM, induction appeared to fail altogether, and so few cells were labeled that the mice were excluded from analysis. Similar failures were never observed in mice treated with 4-OHT.

Because tau stabilizes actin (Fulga et al., 2007), and actin stabilization inhibits mitochondrial localization of DRP1, we postulated that tau exerts its effects on mitochondrial structure and function via excessive actin stabilization. To test our hypothesis directly, we destabilized actin in the presence of tau and monitored the effects on mitochondrial morphology, DRP1 localization, and neurotoxicity. Volasertib order To destabilize actin, we expressed the

actin severing protein gelsolin (Yin and Stossel, 1979) using a UAS-gelsolin transgene. We first confirmed that expression of gelsolin reduces F-actin levels by staining whole-mount brain preparations with rhodamine-phalloidin ( Figure S5). Overexpression of gelsolin reduces mean mitochondrial length, rescues neurodegeneration, and decreases ROS production in tau transgenic neurons ( Figure 5A). We also find increased localization of DRP1 to mitochondria when gelsolin is coexpressed with tau ( Figure 5B, arrowheads) and reduced incidence

find more of elongated mitochondria lacking DRP1 association ( Figure 5B, arrows). These findings overall provide strong evidence that tau blocks DRP1 localization to mitochondria through its influence on the actin cytoskeleton. We wondered if actin stabilization might play a more general role in controlling the subcellular localization of DRP1. We thus examined the localization of DRP1 and F-actin in Cos-1 cells, an immortalized mammalian fibroblastic cell line. Visualization of F-actin using rhodamine-phalloidin and endogenous DRP1 by immunofluorescence with a DRP1 antibody reveals colocalization of DRP1 with actin stress fibers (Figure S6A). We next examined the effects of actin stabilization on DRP1 and mitochondria in these cells, using transient transfection of the actin-stabilizing protein transgelin (Shapland et al., 1993). We first confirmed that expression of transgelin increases CYTH4 levels of F-actin by staining with rhodamine-phalloidin (Figure S6B). Visualizing DRP1 by immunofluorescence and mitochondria by transfection of mitochondrially directed

red fluorescent protein (mitoRFP), we find that in control cells mitochondria are round or modestly tubular, and DRP1 colocalizes with mitochondria (Figure 6A, control, inset). In contrast, in transgelin-expressing cells mitochondria are elongated, and DRP1 shows less mitochondrial colocalization (Figure 6A, transgelin, inset). Quantitative analysis reveals a significant increase in mean mitochondrial length in transgelin-expressing cells compared with controls (Figure 6A, graph). Three-dimensional reconstruction of confocal fluorescence Z-stacks verifies mitochondrial elongation in response to transgelin transfection (Movies S5 and S6). Signal intensity profiles confirm loss of association between mitochondria and DRP1 following transgelin expression (Figure S6C). We next assessed whether these changes in mitochondria morphology correlate with a disruption of mitochondrial bioenergetics.

3). We speculate that the synergy between IL-4 and IL-10 is probably associated with fibrosis contributing to host survival and maintenance of infection. Studies have shown the involvement of cytokines produced by TH2 lymphocytes as IL-4, IL-5 and IL-13 in formation of granuloma in Schistosoma mansoni infection and liver fibrosis progression ( De Jesus et al., 2004). The macroscopic changes observed in the livers were consistent with the expression Selleckchem BMN 673 increased of IL-10 and IL-4. It occurred mainly in livers from animal with fibrosis, necrosis, hemorrhage, and duct calcification and hyperplasia. We demonstrated that a TH2-polarized response predominates in chronically infected

animals, suggesting that maintenance of natural infection is associated with elevated IL-4 and IL-10 levels. Besides the cytokines analyzed, it is important to note that TGF-B has also been demonstrated as an important factor in immunomodulation and probably the establishment of fibrosis in animals infected with F. hepatica ( Haçariz et al., 2009). What can be observed in cattle naturally infected by F.

hepatica is a balance between the expression levels of IFN-γ, IL-4 and IL-10 in the liver tissue confirming the predominance of TH2 response in naturally infected animals from an endemic area. This balance aids in anti-parasite defenses, “monitoring” fascioliasis second progression and survival of the vertebrate host, which can remain in continuous contact with these parasites for prolonged periods of time. Together, Alectinib these results are important to complement previous works indicating that new researches should be made to evaluation of role IL-4 and IL-10 in F. hepatica infection. This study was funded by Brazilian National Council for Scientific and Technological Development (“Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico” – CNPq) and Pro-Dean for Research (“Pro-Reitoria de Pesquisa”) of UFMG. “
“Lippia gracilis Schauer

(Verbenaceae) is a deciduous branched shrub able to grow to approximately 2 m. This plant is a species of the vegetation typical of a well-drained, semi-arid region in northeastern Brazil. Its aromatic leaves and flowers constitute the plant medicinal components, composed of tissues from which the essential oil is extracted. The plant oil exhibits antimicrobial activity due to its high content of carvacrol and/or thymol ( Pessoa et al., 2005). An Active Germplasm Bank of L. gracilis is maintained at the Federal University of Sergipe and plant material is available to supply cuttings for production of plantlets needed for large scale cultivation and essential oil production. Generally, members of the Lippia genus have similar chemical composition, with some compounds present in several species.

These may include positive cues promoting synapse formation INCB024360 molecular weight on CA3 neurons and negative cues preventing synapse formation on CA1 neurons. We do not rule out the possibility that the number of correct synapses is refined over time through other mechanisms. In addition it is important to note that although we always

observe a highly significant bias toward correct target innervation, we also detect incorrect synapses in culture that are not normally found in the brain. This likely reflects the fact that the brain uses several mechanisms (i.e., axon guidance, specific target recognition, synapse elimination) to ensure that neural circuits form with high fidelity. The formation of specific classes of synapses requires communication between two neurons. For this reason transmembrane cell adhesion molecules that interact with the extracellular environment and transmit information inside the cell are attractive candidates for mediating specific synapse formation. The classic cadherin gene family consists of approximately 20 members, and their differential expression in the brain has raised interest in the

possibility that cadherin-mediated interactions play an important role in synaptic specificity (Arikkath and Reichardt, 2008 and Bekirov et al., 2002). However, much of our understanding of the role of cadherins at synapses is based on N-cadherin, which is broadly expressed and appears to have a general Selleckchem LGK974 role in modulating synaptogenesis, spine formation, and plasticity in response to activity (Arikkath and Reichardt, 2008, Bozdagi et al., 2004, Bozdagi et al., 2010, Mendez et al., 2010, Saglietti et al., 2007 and Togashi et al., 2002). N-cadherin is also involved in earlier events including axon guidance and laminar targeting (Inoue and Sanes, 1997, Kadowaki et al., 2007 and Poskanzer et al., 2003), and DG axons respond differentially to N-cadherin versus cadherin-8 (Bekirov et al.,

2008). Despite extensive analysis of N-cadherin function, the role of most other cadherins in synapse formation remains unknown. Cadherin-9 is unique because it is the only cadherin with highly specific expression in DG and CA3 neurons. We found that cadherin-9 is homophilic, localizes to mossy fiber synapses, and is specifically required for formation of a subset of synapses (DG synapses) in culture and in vivo. Non-specific serine/threonine protein kinase To our knowledge, this is the first direct evidence that a cadherin regulates the differentiation of a specific class of synapses. Hippocampal neurons express multiple cadherins and, therefore, it is possible that different kinds of hippocampal synapses are specified by a unique cadherin or combination of cadherins. Cadherins participate in both homophilic and heterophilic interactions, and this feature increases the diversity of synapses that may be regulated by individual cadherins (Patel et al., 2006, Shimoyama et al., 2000 and Volk et al., 1987).