54,55 The reported frequency of mutations in exon 11 varies from 20% to 92%, depending on the type of material (frozen or formalin fixed) and the technique used.8,14,18,31,32,33,51,56,57 Most of the mutations are located between codons 556 and 560, next with deletions and insertions prevalently affecting codons 557�C559 and point mutations affecting codons 559 and 560.8,24,49,51,52,53,58,59,60 Internal tandem duplications are prevalently found towards the end of the exon (codons 576�C580).52 The type of mutation is apparently related to the prognosis, with deletions behaving more aggressively in comparison with insertions and point mutations,8,18,29,58,61,62,63 and to the risk classification. Exon 9 (extracellular domain) The frequency of this mutation is described in 5�C18% of cases, depending on the series.

18,24,28,49,53,64,65,66,67,68,69,70,71,72 It occurs mainly at codons 501�C502 and is represented by duplication�Cinsertion. It is associated with small intestinal localization and aggressive behaviour.18,24 Its mechanism probably affects an antidimerisation motif in the extracellular domain. Exon 13 (kinase I domain) This rare mutation, affecting codon 642, occurs in 0.8�C4.1% of cases.1,3,35,49,64,65,66,70,71,73,74 It is associated with resistance to treatment with imatinib. Exon 17 (activation loop) The activating mechanism of these rare mutations (0.6% of cases)18,33 affecting codons 820 and 822, is unclear. A mutation occuring at codon 817, highly activating and frequently observed in other tumours (mastocytosis, acute myelogenous leukaemia), was never observed in GISTs, implying that the transforming mechanisms in the genesis of GIST are different from those of other tumours.

18,23 Mutation in the pdgfra gene They are observed in 7�C12% of cases,18,20,29,45,49,50,51 occurring more often in exon 18 (activation loop) and rarely in exons 12 (juxtamembrane domain) and 14 (kinase I domain). pdgfra Mutants are prevalently epithelioid, located in the stomach and show weak or no immunohistochemical reactivity for KIT,18,20,29,45,49,50,51,75,76 but are functionally similar to kit mutants. The mutations occur in homologous domains, and activation of the downstream signalling pathways seem to be largely similar in the two mutant subtypes.77 Some degree of difference in gene expression may exist, but these data need confirmation in larger series.

78 Exon 18 (activation loop) Mutations occur at codons 842�C849. Some of them (D842V, RD841�C842KI and Carfilzomib DI842�C843IM) have shown considerable resistance to treatment with imatinib.45,48,49,79 Exon 12 (juxtamembrane domain) Mutations occur at codons 561�C571 and are associated with good response to imatinib.18,48,49,50 Exon 14 (kinase I domain) A single rare mutation is described (N659K). It showed in vitro sensitivity to imatinib that is comparable to that observed in kit exon 13 mutants.

Cells were pelleted and resuspended in 3 ml DMEM- 10% FBS-100 ��g DNaseI-10 ��M Y27632 (Sigma, St. Louis, MO). Verification that the cell suspension was dissociated into single cells was performed by placing 200 ��l of the preparation into a well of a 96-well plate and observing the cells under a fluorescent microscope EMD 1214063 as well as under bright-field optics. Quantification of cells expressing distinct Sox9-EGFP levels (High, Low, and Sublow or Negative) was performed by using a Cyan flow cytometer, Summit v4.3 software, and parameters previously described (21). Propidium iodide staining (Sigma, St. Louis, MO) was used to gate out dead cells. Immune cells were excluded by forward-side scatter gating and doublets were discriminated by use of both forward scatter and side scatter height-width plots.

For each flow cytometry run, the proportion of each cell population at each time point after radiation was compared with the proportion of the respective cell population in nonirradiated control mice studied in the same run. Tissue Harvest for FACS of Different Sox9-EGFP-Expressing Cell Populations These studies focused on nonirradiated mice and mice at 5 days after irradiation when histology revealed maximum crypt regeneration/cell proliferation. FACS was used to isolate cells for testing in the ISC/organoid culture system and for gene microarray. For these analyses, 6- to 10-wk-old nonirradiated Sox9-EGFP mice or Sox9-EGFP mice at day 5 after abdominal radiation were euthanized with a lethal dose of Nembutal and the entire jejunum was placed on ice and processed for IEC isolation and dissociation into single cells as described above.

Prior to FACS, the dissociated cells were sequentially passed through 100-��m, 70-��m, and 30-��m filters. The cells were pelleted and placed in 3 ml Advanced DMEM F12�C10% FBS-100 ��g DNaseI-10 ��M Y27632 (Sigma). Sorting of Sox9-EGFP High, Low, Sublow, and Negative cells was performed by using a MoFlo XDP FACS machine (Dako/Cytomation, Carpinteria, CA) and Summit v4.3 software. Dead and immune cells were excluded by forward-side scatter gating and doublets were discriminated by using both forward scatter and side scatter height-width plots. For each sort of a preparation of dissociated cells from an irradiated mouse, gates delimiting the distinct Sox9-EGFP populations were defined by use of a nonirradiated control littermate sorted in the same run.

Postsort analysis indicated that the different Sox9-EGFP cell populations were correctly isolated on the basis of EGFP status (Fig. 1A). In addition, postsort efficiencies were assessed by flow cytometry after each run for each sample to verify Carfilzomib minimal contamination by other Sox9-EGFP cell types (Fig. 1B). Results demonstrated high postsort efficiencies and negligible cross-contamination of Sox9-EGFP High, Low, Sublow, and Negative cells with other cell populations (Fig. 1B). Fig. 1.

2a). Compared Rucaparib Sigma to normal animals, M2 mRNA relative quantities were increased in all rabbits displaying baroreflex dysfunction and vagal hyperreactivity (Fig. 2b). Figure 2 M2 muscarinic receptor gene expression in peripheral mononuclear white blood cells from normal (N) and vagal hyperreactive (H) rabbits. AchE gene expression and enzyme activity In an attempt to better characterise vagal disorders in H rabbits, we looked at whether AchE gene expression and enzyme activity were also changed in these animals. Vagal hyperreactive rabbits displayed an AchE mRNA amplification ratio of 3.6 versus normal rabbits (Fig. 3a,b); this was associated with twice the enzyme activity in erythrocytes (Fig. 3c).

When AchE was blocked by intravenous administration of neostigmine, a specific AchE inhibitor, H rabbits displayed massively increased bradycardia following PNE injection (up to 10-fold higher in some individuals; Fig. 4). These data confirmed that AchE enzyme activity was increased in the hearts of these animals. Taken together, the findings showed up-regulation of AchE in H rabbits. Figure 3 AchE cardiac gene expression and enzyme activity in erythrocytes from normal (N) and vagal hyperreactive (H) rabbits. Figure 4 Effect of AchE enzyme blockade in normal (N) and vagal hyperreactive (H) rabbits. Evolution of vagal disorders with age As a next step, age-dependent changes in vagal disorders and their functional consequences on R-R intervals were assessed in normal and hyperreactive rabbits. Blood samples were collected at the ages of 5 and 7 weeks.

The M2 expression decreased between the 5th and the 7th week of age in rabbits with normal vagal responses, whereas it remained unchanged in hyperreactive animals (Fig. 5a). In contrast, the AchE expression remained stable in N rabbits but increased significantly in H rabbits within the same period (Fig. 5a). Consequently, the M2/AchE ratio was similar whatever the age and the strain (Fig. 5a). Figure 5 Age-dependent changes in R-R interval and M2 muscarinic receptor and AchE gene expression in peripheral mononuclear white blood cells from normal (N) and vagal hyperreactive (H) rabbits. Functionnal disorders appeared after 7 weeks of age: R-R interval duration was similar in all rabbits up to 7 weeks and then increased in H rabbits, while remaining stable in N animals (Fig. 5b).

Despite SD error-bars appear overlapping on this figure, data were significantly different (P=0.0179 at 8�C11 weeks and P=0.011 at 12�C14 weeks compared to values observed at 7 weeks). Mortality study Mortality has been assessed in Anacetrapib a population of 2150 normoreactive rabbits and 385 hyperreactive animals. In hyperreactive animals, the altered changes in M2 muscarinic receptor and AchE expression were associated with an abnormally high mortality rate between 5 to 7 weeks of age: 52% in H rabbits versus 13% in normal rabbits. Mortality was higher in male than female (57% versus 43%).

4 Flooding of mosquito habitats can introduce RVFV into domestic animal populations by the production of vertically infected Aedes mosquitoes (Figure 1). Epizootic/epidemic cycles are driven by the subsequent elevation of various Culex mosquito populations, which serve http://www.selleckchem.com/products/Erlotinib-Hydrochloride.html as excellent secondary vectors if immature mosquito habitats remain flooded long enough.5 On the basis of this previous research, we have developed a monitoring and risk mapping system6,7 that uses a variety of satellite measurements including sea surface temperatures, outgoing longwave radiation, rainfall, and landscape ecology using the normalized difference vegetation index (NDVI). The measurements represent the total variety of climate and ecological drivers that would lead to conditions associated with the emergence of RVFV vectors resulting in episodic patterns of epizootics/epidemics through time.

These data are input into an RVF prediction system to map in a dynamic manner areas at potential risk for RVF activity.6 This system operates in near real-time to monitor RVF risk on a monthly basis and offers the opportunity to identify eco-climatic conditions associated with potential vector-borne disease outbreaks over large areas, and has been in operation for the last 10 years.7 The system predicted conditions likely to lead to an RVF outbreak in East Africa in September 2006, 3 months before confirmation of disease transmission by the end of November 2006 and human RVF cases mid-December 2006.

8 In this work we focus on the assessment of the predictions with regard to the clusters of outbreaks in East Africa (Kenya, Somalia, and Tanzania): September 2006�CMay 2007; Sudan: May 2007�CDecember 2007; and Southern Africa and Madagascar: September 2007�CMay 2008. The dates mentioned previously represent the time periods when elevated rainfall occurred at least 2 to 3 months before reported RVFV activity (Figure 2). In all of the regions examined except for Madagascar, most of the areas where RVF cases were reported received in excess of 200 mm of rainfall during the outbreak period with the highest excess rainfall occurring in East Africa with amounts up to +400 mm. Details of the setup and implementation of the monitoring and prediction system have been presented in previous works.3,6�C8 Figure 1.

Endemic (on left) and epidemic (on right) life cycles of Rift Valley fever involving close association between heavy rainfall conditions, vector Aedes and Culex mosquitoes, domestic animals, Entinostat and humans. The epidemic cycle is precipitated by excessive … Figure 2. Cumulative rainfall anomalies for (A) East Africa: September 2006�CMay 2008. (B) Sudan: May�CNovember 2007. (C) Southern Africa: September 2007�CMay 2008. (D) Madagascar: September 2007�CMay 2008. Except for Madagascar, all … RVF risk mapping prediction and assessments.

32 This technique has been used in cancer patients,33 and has been found to raise T cell responses to HCV in chimpanzees.34 Another question is which antigens should be used. Ideally, the antigen should be highly expressed in infected cells, and selleck chem Paclitaxel represent a well-conserved viral region so that the vaccine-primed T cells will recognize endogenous virus. A recent meta-analysis suggested that the best antigens to use in a protective vaccine would be the structural antigens.35 Since these are often highly variable and may be less suitable in therapeutic vaccines. The most conserved HCV genes are the core, nonstructural (NS) 3 and NS5B genes,7 which suggests that these are suitable for inclusion in therapeutic vaccines.

In the current study, we initiated a small proof-of-concept study to investigate whether a therapeutic HCV vaccination has an impact on the immune response and whether vaccination has any influence on a subsequent standard-of-care (SOC) treatment in patients with chronic HCV infection. Results Safety and tolerability of DNA vaccination delivered by in vivo EP All patients tolerated the vaccine injections well and no major side effects were noted. Within a minute after the vaccine injection, EP was performed at the same site. The correct delivery of the EP pulses could easily be discerned by two small muscle twitches in the vaccinated arm. A short-lasting pain was recorded that waned within a few minutes. Subjects receiving the EP procedure described the experience qualitatively as ��leaving a small feeling of having been hit��. The perceived pain level has been illustrated in Figure 1.

During and after the EP procedures, patients stayed in the hospital for 2 hours. No major adverse event beside the transient pain was noted. In vivo EP was the cause for the transient local pain. The pain was immediate at administration of the electrical pulses. On a relative 10-level pain scale the in vivo EP gave scores ranging from 2 to 8 whereas the DNA injection gave a score from 0 to 5 (Figure 1; P < 0.01, Mann�CWhitney U-test). The perceived pain did not increase with repeated doses of the DNA vaccine (Figure 1). No significant change in blood chemistry was noted related to the treatment (data not shown). No severe adverse event was seen. Figure 1 Pain levels recorded immediately after in vivo electroporation (EP) given in relation to the pain level related to the DNA injection.

Values are summarized from all four treatments and have been given as the mean pain level estimated using the visual … DNA vaccination induces a transient HCV-specific AV-951 T cell activation The second endpoint of the study was immunogenicity. This was determined both on the B and T cell level. The levels of anti-NS3 IgG were quantified in all samples. The mean endpoint titers did not change significantly during the vaccinations due to high levels of pre-existing antibodies in all patients (data not shown).

Constipation with occasional diarrhea was also commonly experienced by these women, reported by approximately one-half on a weekly basis and by 86.2% at least once per month. TABLE 4 Proportion of women reporting individual symptoms on a weekly and monthly inhibitor DZNeP basis For each symptom, the majority of women rated its usual severity as ��moderate�� (Figure 2). The percentage of sufferers rating each symptom as ��severe�� ranged from 13.7% in the case of bloating, to 28.0% and 31.2% with reference to constipation with occasional diarrhea and abdominal pain, respectively. Overall, 10.4% of respondents rated all their symptoms as ��mild�� and 34.8% of women had one or more ��severe�� symptoms. Figure 2) Usual severity of symptoms. The percentage of women experiencing each of the symptoms does not total 100%.

A small percentage of women were either unable to rate the severity of their symptoms or did not experience an individual symptom. The inclusion … Over 60% of women experienced constipation for more than 10 years, compared with one-half or fewer who experienced bloating, abdominal discomfort, abdominal pain or constipation with occasional diarrhea for more than 10 years (Table 5). However, these differences were not statistically significant. TABLE 5 Duration of specific symptoms On average, 35.4% of women experiencing chronic lower GI symptoms reported that their symptoms were triggered or exacerbated by food and 15.7% reported their lifestyle as the main trigger (patients could cite more than one trigger). However, 53.4% stated that ��anything in particular�� could trigger their symptoms.

Impact of dysmotility and sensory symptoms As many as 63.7% of women experiencing abdominal pain stated that they were ��bothered quite a bit�� or ��extremely bothered�� by this symptom. The corresponding values for those experiencing abdominal discomfort, bloating, constipation or constipation with occasional diarrhea were 53.5%, 46.7%, 47.5% and 57.7%, respectively. Over the past three months, work and social activities of many participants were disrupted by their symptoms. Of the women questioned, 28.8% stated that they were less productive at work or at school (an average of 8.9 occasions). In addition, 13.2% of respondents had missed work or school (an average of five occasions) and 24.7% had missed or were late for a social engagement (an average of 3.

3 occasions). Similarly, 15.3% of women had missed or were late for Carfilzomib an appointment (an average of 3.4 occasions), 9.6% were late for work or school (an average of 5.5 occasions) and 14.7% had left work or school early (an average of four occasions) because of their lower GI symptoms. Almost all participants (97.8%) had made lifestyle changes to cope with their symptoms, such as trying to get ��enough�� sleep (80.1%), wearing looser clothing (72.1%), avoiding certain foods (71.2%), increasing the amount of exercise (66.3%) and fibre intake (65.1%), and avoiding caffeine (30.9%).

Table 1Number of links (first number) and unique genes (second number) at different FBS (final Bayesian score) cutoffs in FunCoup, where c is the corresponding confidence then value of functional coupling.The proteins with the highest connectivity are mainly related to fundamental cellular processes such as protein synthesis and degradation, translation, and transcription (Table S2). Many of them are involved in multiple processes. The most connected protein in our chicken network is the RA-related nuclear protein (RAN). Due to its various functions in nuclear transport and cell cycle regulation, it acts as a major hub with a host of other proteins. Interestingly, RAN is highly differentially expressed between male and female chicken (i.e.

, sex biased) in the gonad (FDR P < 10?4 in the adult), which is actually less common for hubs as we show in the following.3.2. Sex Bias Depends on Network ConnectivityIs there a dependency between sex bias and network connectivity? To answer this question, we first grouped the genes in three sex bias categories: male biased, female biased, and unbiased. For this we used the MWT statistic of differential expression with an FDR P value cutoff of 0.1. This was done for all four tissue/stage conditions: the embryonic and adult gonad and brain. The number of sex-biased genes in the network for each category is shown in Table 2. Remarkably, the embryonic brain contained almost no sex-biased genes and was therefore left out of this analysis. The adult brain had more sex-biased genes, but these still represented only 3% of the genes in the network.

In contrast, the gonad abounded with sex-biased genes in the network: 43% in the embryo and 82% in the adult.Table 2Number of sex-biased and unbiased genes separated by the cut-off FDR<0.1 according to MWT.Sex-biased hub genes were thus frequent in the gonad, but not in the brain, and this may be due to the fact that the male and female gonads have extensive sex-specific functions, while the brain consists of many different tissues of which only small fractions of our microarray samples may be affected by the sex. The sex-specific expression signal in the brain will therefore be diluted by the nonaffected tissues until it is no longer statistically significant. Finer-scale analysis of specific brain tissues might reveal more dimorphism in gene expression, particularly those regions related to vocalization differences between male and female birds [32] or reproductive behavior [33].

We calculated Spearman’s rank correlation coefficient between FDR values from differential expression analysis and node degree (i.e., the number of connections a gene has in the network), for AV-951 each tissue/stage combination. As can be seen in Table 3, all but one of the sex-biased categories in the gonads had a significant positive correlation at FDR P < 0.

In vitro experiments showed that piperine (25�C100��M) significantly stimulated ��-glutamyl transpeptidase selleck chem Sunitinib activity, enhanced the uptake of radiolabelled l-leucine, l-isoleucine, and l-valine, and increased lipid peroxidation in freshly isolated epithelial cells of rat jejunum. In the presence of benzyl alcohol, an enhanced ��-glutamyl transpeptidase activity due to piperine was maintained. These results suggested that piperine may interact with the lipid environment to produce effects which lead to increased permeability of the intestinal cells [12].4. Herbal Bioenhancers4.1. PiperinePiperine, the major plant alkaloid present in P. nigrum Linn (Black pepper) and P. longum Linn (Long pepper), has bioavailability enhancing activity for some nutritional substances and for some drugs [20].

It has been used extensively as a condiment and flavoring for all types of savory dishes. Piper species have been used in folklore medicine for the treatment of various diseases, including seizure disorders [21]. Piperine is known to exhibit a variety of biological activities which include anti-inflammatory activity [22, 23], antipyretic activity [24], fertility enhancement [25], antifungal activity [26], antidiarrhoeal activity [27], antioxidant activity [28�C33], antimetastatic activity [34], antithyroid activity [35, 36], antimutagenic activity [37�C40], antitumor activity [39, 41, 42], antidepressant activity [43�C45], antiplatelet activity [46], analgesic activity [47], hepatoprotective activity [48], antihypertensive activity [49], and antiasthmatic activity [50].

Piperine exhibits a toxic effect against hepatocytes [51] and cultured hippocampal neurons [52], reproductive toxicity in swiss albino mice [53], and immunotoxicity [54].Piperine reduces the aflatoxin B1-induced cytotoxicity and micronuclei formation in rat hepatoma cells in concentration-dependent manner. It is capable of counteracting aflatoxin B1 toxicity by suppressing cytochromes P450-mediated bioactivation of the mycotoxin [55]. Inhibition of aflatoxin B1-induced cytotoxicity and genotoxicity in chinese hamster cells by piperine was reported by Reen et al. [56]. A significant suppression (33.9�C66.5%) in the micronuclei formation induced by benzo(a)pyrene and cyclophosphamide was reduced following oral administration of piperine Dacomitinib at doses of 25, 50, and 75mg/kg in mice [32].Piperine modulates the oxidative changes by inhibiting lipid peroxidation and mediating enhanced synthesis or transport of glutathione thereby replenishing thiol redox [57].

..4. Discussion Symptoms and selleck chem Imatinib signs of dengue fever include sudden onset of high fever, retroorbital headache, rash, joint and muscle pain, some degree of hemorrhage, low platelet counts and liver damage. Several immunodeficient animal models have been described to study different aspects of the disease [7, 9, 10, 14]. Several animal models have been used to study the disease, including immunocompetent [6, 8] immunocompromised [7, 9] and humanized mice [15]. However, it is well known that the immune system combats an invading pathogen by a very complex interplay between the many different components of this system; thus, important interplay between those components can be masked by the immunodeficiency induced in those animals, leading, in some cases, to misleading conclusions.

In this study, we have established an experimental model of DENV infection using immunocompetent C57BL/6 mice, which showed some signs of DF similar to those observed in human. Moreover, the animals became viremic and the virus spread to several organs.Approximately, one-third of DF patients may have mild hemorrhage manifestations, which may be related to the low platelet count [5]. Then, platelet count is a useful marker to be evaluated in an animal model during DENV infection. In that sense, our C57BL/6 mice model showed a significant thrombocytopenia between seven and ten days after DENV-1 infection. In other studies, H. C. Chen, (2007), and Y. T. Yen, (2008), have also shown that C57BL/6 mice infected with DENV-2 showed a significant thrombocytopenia and severe systemic hemorrhage when a very high viral dose (2 �� 109PFU) was used for the infection, but no hemorrhage was observed at lower viral dose (4 �� 107PFU).

In our study, we did not analyze in more details the presence of hemorrhage, but macroscopic hemorrhage was observed in the spleen of the infected animals.After the mosquito biting, human skin dendritic cells (Langerhans cells) are one of the first targets for DENV infection [15�C18]. Then, the virus spread systemically infecting several organs. In that sense, DENV has been found in different Anacetrapib organs such as skin [19], liver [20�C22], spleen [21�C23], lymph nodes [21, 22], kidney [22, 24], bone marrow [22, 24], lung [22, 24], thymus [25], and brain [26]. The animal model analyzed in our study showed the presence of viral genome in kidney, liver, spleen and brain after infection. Although liver is not a major target organ, several studies have demonstrated elevated liver enzyme levels in human serum indicating the damage of this organ after DENV infection [27�C30]. Hepatitis associated with dengue fever is also characterized by moderate hepatocytes necrosis, microvesicular steatosis and cellular infiltration [31�C33].

Results and Discussion3.1. Total Number/Mass Concentration of Particle Emitted from CombustionThe results are shown in enzyme inhibitor Table 2. The total number of particle emitted from combustion of lignite, rice husk, and bagasse are 3.4 �� 103, 1.6 �� 104, and 1.51 �� 105particles/cm3 ? kgfuel, respectively, while total mass of particles are 12.2, 8.0, and 6.5mg/Nm3 ? kgfuel. These results indicate roughly that combustion of low bulk density fuel may be one of the causes to generate higher the emitted particle (compared to lignite). Table 2Total number/mass concentration of particles.However, comparison with 1.8 �� 1013, 1 �� 1013, 1.7 �� 1013 particle/kg released from combustion in self-built burning stove of wheat straw, corn straw, and rice straw, respectively [13]. It seems that the low of density fuel (i.

e., rice husk or bagasse) may not be a priority concerned with high emission of PM but the combustion technology or operating condition seems more importance. Other interesting is that bagasse and rice husk have higher volatile yield than coal, therefore, the main combustion process is marked by devolatilisation rate of fuel and homogeneous (gas-phase) reaction dominated, which later favours particle formation via gas-to-solid pathway (e.g., condensation). According to this phenomenon, this could be observed from the reverse relationships between particle number and particle mass concentration. For instance, most prevailing size of particle of bagasse combustion is at dp 70nm of 80% cumulative of total number of particle.

Because this high content of submicron particles is less significant to contribute the overall mass loading, thus low mass concentration does. In addition, low mass of emitted particle also indicates the lower particle density of residues. 3.2. Total Number/Mass Concentration of Particle Emitted from Cofiring of Lignite and Rice HuskCofiring of lignite and rice husk was performed under various mass fraction and ratio of overfired air to total air and results are shown in Table 3. Table 3Total number/mass concentration of particle.It can be seen that cofired lignite and rice husk result in increase of both particle number and mass concentration compared to burning of either lignite or rice husk. This synergy effect could be from the difference in fuel properties and physical which needed further investigation and analysis.

While mass fraction concentration has affected to PM emission, an increase in lignite mass concentration leads to decrease in PM emission. However, total number/mass concentration of particle is decreased dramatically at overfired air to total air ratio of 0.1. The result of particle number Entinostat of fuel mixture (8.7 �� 103) is in between those of lignite and rice husk (3.4 �� 103 and 1.6 �� 104) but mass concentration is much lower. This could be said that PM emission at this condition probably is very fine particle. 3.3.