First, a more complex spectrum of treatment changes among women, driven by more symptoms/side effects attributed to ART. Second, less intake of protease inhibitors among women, potentially mitigating or reversing treatment-associated lipodystrophy and metabolic complications by omitting these components of ART [27]. Third, the disproportionate http://www.selleckchem.com/products/Erlotinib-Hydrochloride.html prevalence of renal dysfunction among men, possibly reflecting a higher exposure to toxicities of ART [28] (in our study, men were taking ART on average 18 months longer than women). Finally, since in the German population women are known to report a higher frequency and severity of symptoms [29], we expected to have similar findings in the population of people with HIV.

However, in our study women and men with HIV did not differ on the frequency and severity of symptoms, which also may indicate that women were more successful in reducing side effects of ART. Sharing lab results -gaps in physician-patient communication? Another imperative discovery of our study was the incongruence between physicians’ and patients’ reports of laboratory abnormalities. Only one of seven participants indicated potentially ART-related laboratory abnormalities that were documented in the physician’s records. This might partially be explained by attribution of laboratory abnormalities to other causes, such as co-infection with viral hepatitis. Some laboratory abnormalities may simply be not avoidable, such as lipid and glucose elevations in patients with a predisposition to develop a metabolic syndrome, or elevated liver enzymes in patients with a hepatitis co-infection.

Clinical significance of laboratory abnormalities depends on the treatment, the treatment experience, and the patient’s vulnerability to develop certain side effects. Since the majority of the laboratory abnormalities were mild, physicians may not have discussed them with their patients or patients did not recall the discussion. A patient’s forgetfulness of laboratory abnormalities may indicate that his/her focus is on benefit rather than on risk of the treatment, weighing long survival with HIV against laboratory alterations such as elevated lipids, liver enzymes, or lactic acidosis. By focusing on HIV, some patients may ignore their individual risk of cardiovascular or liver disease.

Thus, Drug_discovery a patient’s “selective memory” may serve as an individual strategy to cope with the disease and its treatment and to avoid panic about minor laboratory alterations. Nevertheless, in the context of ART, abnormalities of liver enzymes, even if they are mild, require further examination of the etiology and may be an indicator of ART-related mitochondrial injury and hepatic steatosis [30,31]. Since potentially ART-related laboratory abnormalities might be amenable to change by treatment modifications, the lack of patients’ awareness of these abnormalities should be addressed.

We examined the effect of PKC activation with the phorbol esters PMA or PdBu on selleckchem hASIC1b currents. PMA and PdBu are cell-permeable diacylglycerol (DAG) analogs that activate classical and novel PKC isoforms. Oocytes were injected with WT hASIC1b, S40A hASIC1b, or S499A hASIC1b to examine the importance of the amino acids that are consensus PKC phosphorylation sites. Acid-activated currents were recorded in the same oocyte before and after 1 ��M PMA or 1 ��M PdBu addition to the bath solution for 5 min. Incubation of the oocyte with PMA decreased IpH4.0 of WT hASIC1b [0.70 (SD 0.20), n = 29, P = 0.0008 by two-tailed paired t-test of IpH4.0 before and after treatment]. PdBu also inhibited WT hASIC1b current [0.67 (SD 0.12), n = 6, P = 0.010 by two-tailed paired t-test].

Incubation of WT hASIC1b-injected oocytes in the bath solution (ND96 pH 7.4 solution) for 5 min without treatment (n = 27), with DMSO (1:1,000, n = 10), or with the inactive PMA analog 4-��-PMA (1 ��M, n = 12) had no effect on acid-activated currents (Fig. 4A). The 95% CIs for the means of the ratio of IpH4.0 after treatment to IpH4.0 before treatment for the WT hASIC1b construct were (0.940, 1.09) for 5 min of no treatment, (0.852, 1.05) for DMSO, (0.856, 1.19) for 4-��-PMA, (0.626, 0.783) for PMA, and (0.544, 0.792) for PdBu. A statistically significant effect of PMA or PdBu on hASIC1b currents was not observed in oocytes expressing S40A hASIC1b [0.87 (SD 0.13), n = 8, P = 0.061 by two-tailed paired t-test for PMA treatment and 0.80 (SD 0.059), n = 4, P = 0.10 for PdBu treatment; Fig. 4B].

The 95% CIs for the ratios of IpH4.0 after treatment to IpH4.0 before treatment for the S40A construct were (0.799, 1.05) for 5 min of no treatment, (0.768, 0.984) for DMSO, (0.922, 1.00) for 4-��-PMA, (0.766, 0.978) for PMA, and (0.7, 0.89) for PdBu. The S499A hASIC1b construct responded to the activation of endogenous oocyte PKC with PMA or PdBu in a similar manner to WT hASIC1b, decreasing by a similar amount [0.64 (SD 0.24), n = 8, P = 0.033 by two-tailed paired t-test for the PMA effect and 0.55 (SD 0.23), n = 3, P = 0.064 by two-tailed paired t-test for the PdBu effect; Fig. 4C]. The 95% CIs for the mean ratios of IpH4.0 after treatment to IpH4.0 before treatment for the S499A channel were (0.914, 1.09) for 5 min of no treatment, (0.645, 1.15) for DMSO, (0.844, 1.01) for 4-��-PMA, and (0.

461, 0.824) for PMA. Fig. 4. PKC activators PMA and phorbol 12,13-dibutyrate (PdBu) reduce acid-activated currents of WT hASIC1b Cilengitide and S499A hASIC1b but not that of S40A hASIC1b. Oocytes were injected with RNA for WT hASIC1b (A), S40A (B), or S499A (C). Acid-activated currents were … Because PKC can have cell type-specific effects (11), we repeated the experiment of activating PKC with PMA in CHO cells transfected with WT hASIC1b-eGFP bicistronic vector. Acid-activated currents were recorded before and after the superfusion of 100 nM PMA in the chamber.

3 to <1.0), an early transient effect of the vaccinations was noted (Figure 2a). The first vaccination consistently induced an increase in relative antibody levels detected by a paired comparison of the samples Seliciclib obtained at week 0 and 2 in all patients (Figure 2a; P < 0.05, Wilcoxon's matched pairs test). The increase was most pronounced in the two lowest dose groups (P < 0.01). Three of the six subjects in the two lowest dose groups with an increase in NS3 antibodies had de novo T cell activation (labeled with an asterisk (*) in Figure 2a). Thus, a limited effect of the vaccination seemingly was observed in the presence of pre-existing antibody levels. Figure 2 HCV NS3/4A-specific immune responses after vaccination.

Immunogenicity of the first two doses of DNA determined as (a) increases in relative antibody levels to NS3 or (b,c) increases in T cell responses to overlapping peptide pools spanning the complete … The presence of HCV-specific T cell responses before, during, and after the therapeutic vaccination was determined as the number of interferon (IFN)-�èCproducing T cells, or spot-forming cells (SFCs) by ELISpot (Figures 2�C5), and the level of proliferation as determined by the level of [3]H-thymidine incorporation (data not shown). In the ELISpot assay, only the responses to nine peptide pools spanning the whole NS3/4A region were used for the statistical comparison to avoid repeated use of the same epitope and to overcome HLA restriction. The number of the IFN-�èCproducing spots seemed to increase after the two first vaccinations when comparing the number of SFCs at week 0, and the same at weeks 2 and 6 (Figure 2b).

Proliferative T cell responses to NS3 or NS4 were detected in 8 out of 12 subjects before or after vaccination (data not shown). In the 167 and 500 ��g dose groups, de novo ELISpot responses appeared in four subjects, and in the highest dose group, one showed appearance of de novo ELISpot responses (Figures 2c and 5). We compared the breadth of the T cell responses to the nine NS3/4A genotype 1a peptide pools by ELISpot 2 weeks after vaccinations with the responses at week 0 (Figures 2c and 5). Before vaccination, a total of 162 peptide pools from 11 of the patients were assayed by ELISpot, and three (2%) were positive (Figures 2c and 5). Altogether, 39 out of a total of 450 peptide pools assayed after vaccination were positive (Figures 2c and 5; P < 0.

0001, Fisher’s exact test). In addition, the frequency of responses to the nine peptide pools significantly increased treatment at Dacomitinib week 2 (12 out of 99, P < 0.05, Fisher’s exact test), week 6 (10 out of 81, P < 0.05, Fisher’s exact test), and week 36 (9 out of 108, P < 0.05, Fisher’s exact test), when compared with week 0 (1 out of 81; Figures 2c and 5). The responses were most pronounced in the 167 and 500 ��g dose groups.

When human MSCs were cultured during 21 days for hepatocytes differentiation, marked differences in www.selleckchem.com/products/Nilotinib.html the levels of CD13, CD49e, CD133, CD166 and VEGFR2 were observed in CM1 and CM2-treated cells. The levels of these markers were significant higher in CM2-treated cells as compared to undifferentiated cells or CM1-treated cells (Figure 1). Figure 1 Differential stem cells markers in undifferentiated and differentiated human mesenchymal stem cells. Table 1 Stem cells/cancer stem cells markers expression in undifferentiated cells at time 0 days. Expression of hepatospecific markers in human mesenchymal stem cells during their differentiation into hepatocytes To compare the potential for hepatic differentiation of both protocols, the expression of hepatospecific genes was evaluated measuring mRNA levels by RT-PCR and protein expression by immunocytochemistry.

The mRNA levels of albumin (ALB), ��-fetoprotein (��FP), ��1-antitrypsin (��1-AT), C/EBP�� and CYP3A5 were strongly induced in human MSCs treated with CM1 or CM2 at 7, 14 and 21 days, compared to undifferentiated cells (UC). Comparatively, at 21 days of differentiation, there are no significant differences between both differentiation protocols. In CM2-treated cells there were not significant differences in the expression of these genes between cells to time 0 and cells after 48 h of treatment. The expressions of albumin and C/EBP were bigger in CM1 than CM2-treated cells while the expressions of ��1-AT and CYP3A5 were increased in CM2 vs. CM1-tretaed cells. The expression of ��FP was similar with both protocols (Figure 2).

Figure 2 The treatments with CM1 or CM2 increase the expression of hepatospecific genes in human mesenchymal stem cells. Expression of hepatospecific proteins, such as albumin, ��1-antitrypsin, ��-fetoprotein and cytokeratin 19 were also immunodetected in cells after 21 days of differentiation with both protocols CM1 or CM2 (Figure 3). Both protocols expressed with similar intensity hepatospecific proteins. Undifferentiated cells at 21 days were negative for the expression of these proteins. PAS stain was positive after treatment with CM1 and CM2 although a higher intensity was observed in cells cultured with CM2. Figure 3 The treatment with CM1 or CM2 induces the presence of hepatospecific proteins in human mesenchymal stem cells.

Role of Wnt/��-catenin pathway and p53 during human mesenchymal stem cells differentiation Cilengitide into hepatocytes Since Wnt/��-catenin pathway plays a main role in the control of differentiation of adult stem cells, confocal microscopy was used to study the subcellular localization of ��-catenin during differentiation into hepatocytes in both CM1 and CM2 (Figure 4A). Immunofluorescence staining demonstrated that ��-catenin localized to the cell membrane or to the peri-membrane region in undifferentiated and CM1 cells after 21 days, while there was no evidence of nuclear translocation.

Unlike TNF-R1, internalisation kinase inhibitor Brefeldin A is not required for TRAIL-induced apoptosis, as the assembly of the TRAIL DISC already occurs at the cell membrane (Kohlhaas et al, 2007). However, internalisation of the ligated receptor has been suggested to play a role in other TRAIL-mediated signalling events such as activation of JNK or NF-��B (Varfolomeev et al, 2005). In our hands, internalisation of TRAIL receptors ligated by agonistic DR4 and DR5 antibodies or by rhTRAIL revealed no significant differences; in all cases, the ligated receptor complex was rapidly internalised. This indicates that the opposing apoptosis-modulatory effect of JNK activation induced by TRAIL or selective DR4/DR5 activation was not due to receptor internalisation but possibly different isoforms of JNK activated by the individual receptors.

Ten isoforms of JNK are known to exist as a result of alternative splicing of the three genes, jnk1, jnk2 and jnk3 (Gupta et al, 1996). Little is known about the role of these isoforms in apoptosis. Overexpression of JNK1��1 increases resistance to vesicular stomatitis virus-induced cell death in 3T3 fibroblasts, whereas overexpression of JNK1��1 and JNK1��1 potentiates cisplatin- and doxorubicin-induced cell death in sarcoma cell lines (Han et al, 2002; Koyama et al, 2006). Previous studies have demonstrated a role for either JNK1 or JNK2 in TNF��-induced apoptosis (Dietrich et al, 2004; Liu et al, 2004). Our results show that the chief JNK isotype activated by DR4 and DR5 is JNK1.

Furthermore, whereas TRAIL-mediated receptor activation led to activation of both the long and short isoforms of JNK1, selective ligation of DR4 or DR5 with cross-linked agonistic antibodies predominantly activated the short JNK1 isoforms (JNK1��1 and/or ��1) and the difference in cell death seen following JNK inhibition may be related to the different JNK1 isoforms, with the short isoforms GSK-3 of JNK1 (JNK1��1 and JNK1��1) transmitting an antiapoptotic signal and the long isoforms of JNK1 (JNK1��2 or JNK1��2) mediating a proapoptotic signal. As a reason why activation of individual TRAIL receptors by agonistic antibodies activates different JNK isoforms from rhTRAIL, it is likely that the agonistic DR4- and DR5-specific antibodies trigger different receptor clustering, or intracellular conformational changes from those induced by rhTRAIL. It is also possible that on rhTRAIL treatment, higher-order heteromeric receptor complexes (receptosomes) including both DR4 and DR5 are formed where the interaction between the death domains of the various receptor trimers allows for the recruitment of more and/or different adaptor proteins.

The resistance development process will be substantially delayed, ultimately leading to enhanced life-span of the novel and existing antibiotics. Such drugs/molecule facilitators should have novel properties such as (1) nontoxic to humans or animals, (2) they should be effective at a very low concentration in a sellckchem combination, (3) they should be easy to formulate, and (4) most importantly enhance uptake/absorption and activity of the drug molecules. This can lead to developing judicious and strategic concentrations of antibiotic with specific bioenhancers to improve availability of the drug for controlling the infectious organisms effectively [2].Herbal bioenhancer is an agent of herbal origin or any phytomolecule, which is capable of enhancing bioavailability and bioefficacy of a particular drug or nutrient with which it is combined, without any typical pharmacological activity of its own at the dose used.

In the 1920′s, Bose, an acknowledged author of ��Pharmacographia Indica,�� reported an enhanced antiasthmatic effect of an Ayurvedic formula containing vasaka (Adhatoda vasica) when administered with long pepper [3]. The term bioavailability enhancer was first coined by Indian Scientists at the Regional Research laboratory, Jammu (RRL, now known as Indian Institute of Integrative Medicine) discovered and scientifically validated piperine as the world’s first bioavailability enhancer in 1979 [4].The concept of bioenhancers of herbal origin can be tracked back from the ancient knowledge of Ayurveda system of medicine. Use of ayurvedic preparation ��Trikatu�� from the period between the 7th century B.

C. and the 6th century A.D., which is a Sanskrit, word meaning three acrids. It refers to a combination of black pepper (Piper nigrum Linn.), long pepper (Piper longum Linn.), and ginger (Zingiber officinale Rosc.), which contains active component piperine, which enhances the bioavailability of drugs, nutrients, and vitamins [2].2. Bioavailability/Bioefficacy-Enhancing ActivityThe term bioavailability or bioenhancing activity is defined AV-951 as ��a substance at a lower dosage level, which in combination with a drug or nutrient provides more availability of the drug by reducing the consumption of the drug or nutrient resulting in enhanced efficacy of the drugs.��The great interests for the improvement of bioavailability of a large number of drugs are (1) poorly available, (2) administered for long periods, (3) toxic, and (4) expensive. Maximizing bioavailability is therapeutically important because the extent of bioavailability directly influences plasma concentrations and consequently therapeutic efficacy. Bioavailability enhancement can make the expensive drugs affordable and reduce the toxic effects by reducing the required dose of drugs.

Therapeutic PossibilitiesThe use of COX inhibitors for posttraumatic neuropathic pain is an established therapeutical intervention. However, given the association that exists between AA derivatives and nerve degeneration and regeneration, the therapeutic selleck screening library modulation of this pathway emerges as a novel strategy aimed at increased motor, sensory, and structural recovery after nerve injury. We summarize some of the potential mechanisms involved in Figure 1. Currently, COX inhibitors, LOX inhibitors, PG analogues, among other drugs, are approved for human use in clinical situations (Table 1). The development of phospholipase modulators and other drugs targeting AA derivatives is an active field of research, which will soon clarify the validity of this therapeutic strategy.Figure 1Proposed theoretical framework.

After nerve injury, AA derivatives produced in neurons and microglia participate in nerve degeneration and regeneration, through local, remote, and molecular pathways. Injury promotes SC infiltration and activation as well …Table 1Clinically approved modulators of arachidonic acid derivatives. The dual function of AA derivatives, as promoters and inhibitors of nerve degeneration and recovery, will complicate the clinical application of these drugs. It is becoming evident that the dual role of neuroinflammation as both injurious and as a promoter of regeneration is a complex one, even at the molecular level. It is most likely that a combination approach will be fruitful, using compounds that optimize regeneration and others that diminish degeneration.

The exact time of administration will also be important, since some of these molecules are expressed early after injury, while others begin to be produced in the late stages after regeneration. Additionally, although the evidence reviewed here concerns primarily peripheral nerve injury, most of the concepts could be applied both to spinal cord and CNS injury. In conclusion, the findings reviewed here point toward a new avenue in the pharmacological treatment of nerve injury, based on a pathophysiologically relevant paradigm (Table 2). The numerous compunds that exist today suggest that clinical studies are warranted.Table 2Selected animal and human therapeutic studies.Conflict of InterestsThe authors declare no conflict of interests.
Acute myocardial infarction (AMI) is one of the most important cardiovascular diseases with large medical expenditures.

Decreased occurrences of AMI have been demonstrated epidemiologically [1, 2]. Through corrections of risk factors and intensive medical management, the mortality of AMI was significantly reduced [3�C6]. Gender differences play an important role in the pathophysiology of AMI. Although coronary plaque rupture with acute thrombosis formation is common pathophysiology AV-951 for men and women, women are usually older than men and associated with a low incidence of AMI, but with a higher mortality [7�C16].

[17] observed that 12 (29%) of 41 patients treated with of BoNT-A for BPH did not experience reduction of prostate volume, yet seven of these men had significant improvement of IPSS and Qmax . These data suggest that BoNT-A may act on the selleck inhibitor dynamic obstructive component of BPH. The neurotoxin was originally thought to act only by inhibiting acetylcholine release at the presynaptic neuromuscular junction [25]. Presently, other mechanisms are known to be involved such as blockage of neuroglandular junctions. It is believed that it also promotes a decrease of norepinephrine release from sympathetic endings, leading to the consequent reduction of alpha-1A adrenoceptor stimulation [26]. Furthermore, in an experimental model using rats, a dose-dependent decrease in the expression of alfa-1A adrenoceptors was demonstrated [27].

This is another possible mechanism affected by BoNT-A treatment, which may promote a decrease in density of alfa-1a adrenoceptors, which are known to be increased several-fold in BPH [28]. Different doses (range from 100 to 300U) have been studied in several series, but there is a lack of consistency in some studies [15�C21]. Some authors suggest that prostate size might influence the dosage [17, 21], but whether larger prostates require higher doses has actually not been tested. Moreover, there is no evidence whether the severity of LUTS influences the optimal dose. To the best of our knowledge, it is the first study comparing two different doses of BoNT-A. The procedure has been performed by transperineal, transrectal and transurethral approaches.

We chose the transurethral route because the vast majority of urologists are trained on cystoscopic procedures and also because it permits direct vision and injection in the transition zone of the prostate. Kuo [21] firstly described the cystoscopic approach using light general anesthesia or sedation. We have demonstrated that it is feasible to perform this technique with local anesthesia. Pain, when present, was mild and no patient required narcotics in the postoperative period. Complications were uncommon in our series and seemed to be related to the urethral manipulation rather than a direct result of BoNT-A injection. Gross hematuria, although conservatively managed, was observed in 2 (5.8%) patients. Transient urinary retention was observed in 2 (5.

8%) patients, and treated conservatively with an indwelling catheter for 5 days. Two patients (5.8%) developed acute prostatitis. As mentioned, these adverse effects seem to be associated with the route of administration of BoNT-A, since they are potential complications of any procedure requiring urethral instrumentation. Series that adopted the transperineal Batimastat or transrectal approach did not report such complications [22�C27]. Given the vascular nature of the prostate, systemic absorption of the toxin could occur.

These values are mainly based on the poor aqueous solubility (13.6��g?g?1 selleckchem in pH = 7.4) of resveratrol [34] which leads to an increase in the drug loaded into microparticles. Similar results were previously reported. A resveratrol entrapment higher than 97.7% was achieved for calcium-pectinate beads prepared by instantaneous gelation of pectin [11]. Resveratrol-loaded nanoparticles showed EE from 78.3 to 91.4% using PCL of Mw? = 65,000g?mol?1 [16]. Vanillin cross-linked chitosan microparticles containing resveratrol revealed a drug entrapment higher than 93.7% [22]. The polymer:drug ratio is also a critical factor during microparticle formation and can influence EE values [35]. The enhancement of resveratrol entrapment was observed when the polyester amount was increased (Table 3).

For PHBV microparticles, EE was increased from 80 to 93% as polymer:drug ratio was improved from 4:1 (20% resveratrol) to 19:1 (5% resveratrol). Resveratrol entrapment varied from 88 to 101% for PCL microparticles when polymer:drug ratio was changed at the same proportion. This effect can be simply due to the greater polymer with respect to the drug amount.3.2. Scanning Electron MicroscopyThe scanning electron micrographs of PHBV/PCL microparticles are shown in Figure 2. Different morphological aspects were observed depending on the polyester used. By SEM, PHBV microparticles were spherical shaped with a rough surface and pores (Figures 2(a)�C2(d)). The presence of pores represents important morphological evidence that can change the drug release process from microparticles [36].

However, PCL microparticles revealed a spherical shape with smooth surface (Figures 2(e)�C2(h)), and no pore were observed. Moreover, formulation M2R20 showed a residual resveratrol onto the microparticles surface. This external drug can be rapidly dissolved into an aqueous medium and provide an immediate-release behavior (burst effect) [23]. This effect has been also observed in other studies related to delayed-release biopolymer systems [37, 38].Figure 2Scanning electron micrographs of PHBV/PCL microparticles: M1R0 (a), M1R5 (b), M1R10 (c), M1R20 (d), M2R0 (e), M2R5 (f), M2R10 (g), and M2R20 (h). Magnifications of 2000x.3.3. Particle Size and Size DispersionThe particle size and size distribution obtained by LDS measurements are indicated in Table 3.

Micrometer-sized particles with mean diameters less than 60��m were obtained. Although these particle sizes do not allow an uptake by intestinal tissue, the oral administration of these microparticles can provide sustained drug effect due to their prolonged bowel transit GSK-3 time [39]. Regarding that a low span value indicates a narrow polydispersity [40], PCL microparticles presented a more homogeneous size distribution when compared to PHBV microparticles.3.4.

1. Energy ConsumptionEnergy consumption is an important source of GHG emissions in the industrial park. As defined in this study, energy consumption processes involved mainly includes fossil fuel combustion, electricity and heat energy next production, and transportation vehicles. The estimation of the GHG emission from above processes refers to the 2006 IPCC Guidelines for National Greenhouse Gas Inventories [22].For the fossil fuel combustion, (1) is given asEGHG,ic=Qic��EFGHG,ic,(1)whereE(GHG,i)c is the emissions of a given GHG by type of fuel (kg), Qic is the amount of fuel combusted (TJ), andEFGHG,ic is the emission factor of a given GHG by type of fuel (kg GHG/TJ).For the electricity and heat energy productionprocesses,if the power comes from the sectors outside the industrial park, the concerned GHG emission can be calculated according to the average energy consumption level of the local sector.

If the power is produced by the enterprise just in the industrial park, the concerned GHG emission can be estimated as zero to avoid the repeated calculation based on the fossil fuelofcombustion process. Equation (2) is presented to calculate the GHG emission from electricity and heat energy productionprocesses asEGHGe=Qe��EFGHGe,(2)where EGHGe is the emissions of a given GHG by electricity or heat (kg), Qe is the amount of electricity or heat produced (kWh or TJ), and EFGHGe is the emission factor of a given GHG by electricity or heat (kg GHG/kwh or kg GHG/TJ).As for the transportation process, it refers to the transportation of the energy resources, raw materials, instruments and equipment from the origin to the industrial park.

The GHG emissions from the transportation process can be estimated from the fuel consumed:ECO2t=��i(Qit��EFCO2,it),(3)whereECO2t is the emissions of CO2 (kg),Qit the fuel consumed (TJ), and EFCO2,it is the emission factors (kg CO2/TJ), or calculated by the distance covered by the vehicles asENH4,N2Ot=��a,b,c,d(La,b,c,d��EFa,b,c,dt)+��a,b,c,dCa,b,c,d,(4)where ENH4,N2Ot is the emissions of CH4 or N2O (kg),La,b,c,d is the distance travelled during thermally stabilized engine operation phase for a given mobile source activity (km), EFa,b,c,dt is the emission factor (kg/km), Ca,b,c,d is the emissions during warm-up phase (cold start)(kg), a is the fuel type (e.g., diesel, gasoline, natural gas, and LPG), Batimastat b is the vehicle type, c is the emission control technology (such as uncontrolled, catalytic converter, etc.), and d is the operating conditions (e.g., urban or rural road type, climate, or other environmental factors).In general, the first approach (fuel sold) is appropriate for CO2 while the second (distance travelled by vehicle type and road type) is suitable for CH4 and N2O.2.2.