Meprin was first discovered in proximal epithelial cells of mouse kidney in 1981 by Beynon et al. (24). In addition, two other groups discovered the same metalloprotease in the early 1980s: Sterchi et al., in 1982, described the enzyme PABA peptide hydrolase (PPH) in microvillar membranes of human small www.selleckchem.com/products/Vorinostat-saha.html intestinal epithelial cells (25), and Kenny et al., in 1987, found the endopeptidase-2 in the microvillar membrane of proximal epithelial cells of rat kidney (26). This metalloprotease is now known as ��meprin.�� There are two evolutionary related isoforms: Meprin�� and meprin��. Both are synthesized as type I transmembrane proteins in the endoplasmic reticulum (27, 28). The membrane anchor of meprin�� is removed intracellularly leading to the secretion of this isoform from cells, whereas meprin�� remains an integral protein of the plasma membrane (29).

However, meprin�� may be retained at the plasma membrane via covalent interaction with the transmembrane meprin�� (30�C32). Meprin�� is expressed in epithelial cells of the healthy colon mucosa where it is secreted apically into the colon lumen (33). In colorectal cancer, meprin�� is released in a non-polarized fashion, leading to its accumulation in the tumor stroma (34, 35). This aberrant secretion of meprin�� into the tumor stroma exposes matrix components and other stromal elements to an increased proteolytic potential (35). Once secreted, meprin�� is activated in vitro and in the gut lumen by the removal of the pro-peptide through trypsin (28). An alternative activation mechanism has been suggested in colorectal cancer.

In colon carcinoma cells (Caco-2), basolaterally secreted meprin�� is activated by plasmin, which in turn, is activated by the fibroblast-derived urokinase-type plasminogen activator (36). Meprin�� has been demonstrated to have pro-migratory and pro-angiogenic effects in colorectal cancer, and thus may be involved in the transition from benign growth (adenomas) to malignant primary tumors (37, 38). We investigated the molecular mechanisms by which meprin�� may influence tumor progression. For the first time we demonstrate that meprin�� is able to shed EGF from the plasma membrane, resulting in the transactivation of EGFR signaling pathway and enhancement of Caco-2 cell proliferation and migration. We also confirm the shedding of TGF�� by meprin��.

EXPERIMENTAL PROCEDURES Antibodies and Recombinant Protein Antibodies specific for total EGFR (monoclonal rabbit antibody) and phospho-EGFR Y1068 (monoclonal rabbit antibody) were purchased Cilengitide from Epitomics (Burlingame, CA); antibodies specific for total ERK1/2 (monoclonal mouse antibody) and phospho-ERK1/2 (polyclonal rabbit antibody) were from Santa Cruz Biotechnology (Heidelberg, Germany). Horseradish peroxidase-linked anti-rabbit and anti-mouse secondary antibodies were obtained from Dako Cytomation (Denmark).

Isotype control (IgG-PE) was purchased from BD pharmingen (BD Biosciences San Jose, CA, USA). Anti-JAK, anti-p53, anti-p21waf, and anti-survivin www.selleckchem.com/products/Bortezomib.html were purchased from Cell Signaling Technologies (Beverly, MA). Anti-beta-actin antibody was purchased from Sigma-Aldrich (St. Louis, MO). The STAT3 inhibitor WP1066 and the JAK inhibitor pyridone 6 were purchased from Merck (Darmstadt, Germany). Neutralising anti-EGFR and anti-IL-6R antibodies were purchased from Millipore (Billerica, MA) and R&D Systems (Minneapolis, MN) respectively. Recombinant glycoprotein gB was purchased from Abcam (Cambridge, UK). Ganciclovir was purchased from Roche (Basel, Switzerland). Cell culture HepG2 cells were obtained from the European Collection of Cell Cultures (ECACC, Porton Down, UK) and PHH from Kaly-Cell (Strasbourg, France).

HepG2 cells were cultivated in Eagle’s Minimum Essential Medium (EMEM) supplemented with 10% fetal bovine serum, 1% non-essential amino acids (Sigma-Aldrich, St. Louis, MO), penicillin (100 IU/ml), and streptomycin (100 microg/ml). PHH were cultivated in serum-free Dulbecco’s Modified Eagle Medium supplemented with L-glutamine (2 mmol/l), insulin (4 microg/ml), dexamethasone (1 micromol/l), and gentamycin (50 mg/l). The PHH were free of HCV, HBV, HIV, and HCMV as determined by highly sensitive PCR and RT-PCR assays (Abbott; Argene). Cell viability assay was performed as previously described [21]. IL-6 production was measured in culture supernatants using an ELISA kit (Human IL-6 Quantikine ELISA kit, R&D Systems, Minneapolis, MN).

Quantification of HCMV titers in cell culture supernatants was performed by real-time PCR as previously described [11]. HCMV infection of HepG2 cells and primary human hepatocytes Cell-free virus stock was prepared by propagating two strains of HCMV, the laboratory strain AD169 and a clinical isolate, HCMV-DB, in MRC5 human fibroblasts as described previously [11], [21]. AD169 is a highly-passaged laboratory strain of HCMV originally isolated from the adenoids of a child [22]. The clinical isolate HCMV-DB was isolated from a cervical swab specimen from a 30-year-old pregnant woman [21]. MRC5 human fibroblasts were cultured in EMEM with 10% FBS, penicillin (100 IU/ml), and streptomycin (100 microg/ml). HepG2 cells and PHH were infected at different multiplicities of infection (MOI) for 2 h at 37��C, washed thoroughly, and covered with fresh medium.

Where specified, cells were treated with ganciclovir (5 microg/ml) during infection with HCMV. Ultraviolet (UV)-inactivated HCMV (UV-HCMV; 1200 microJ.cm?2, 15 min) was used as control. Supernatants were clarified by centrifugation and stored at ?80��C Cilengitide until use. Virus titers were determined by plaque-forming assay in MRC5 human fibroblasts as described previously [23]. RT-PCR assay Briefly, total RNA was extracted from HepG2 cells with RNeasy mini kit (Qiagen, Hilden, Germany).

Melatonin (N-acetyl-5-methoxytryptamine), the major research use secretory hormone of the pineal gland (42), is also secreted from several extrapineal tissues, such as gastrointestinal tract and liver (21). Melatonin is synthesized from serotonin by two enzymes, serotonin N-acetyltransferase or arylalkylamine N-acetyltransferase (AANAT) and acetylserotonin O-methyltransferase (ASMT) (23). There are three melatonin membrane receptor subtypes, MT1, MT2, and MT3 (found only in nonmammals), through which melatonin exerts its multiple effects (59). Melatonin receptors are present in the central nervous system, as well as peripheral tissues, including small intestine and hepatocytes (36, 40). There is growing evidence that cholangiocytes secrete a number of neuroendocrine factors (e.g.

, dopamine and serotonin) regulating the neoplastic growth of cholangiocytes by autocrine mechanisms (1, 10). A number of studies have shown that melatonin secretion is impaired in patients suffering from breast cancer, endometrial cancer, and colorectal cancer (49, 53, 58), suggesting that reduced melatonin levels are linked to higher incidence of several types of tumors. However, no data exist regarding the role of melatonin as an autocrine regulator of CCA growth. In the liver, melatonin protects against cholestatic oxidative stress in rats during cholestasis (13) and reduces the regeneration and stimulated the apoptosis of hepatocytes in hepatectomized rats (26). In support of the specificity of the present study, we have shown that melatonin inhibits biliary hyperplasia in cholestatic rats (43).

Melatonin is also chemopreventive against O. viverrini-induced CCA by reducing oxidative and nitrosative DNA damage in rabbits (32). On the basis of this background, we hypothesize that 1) in CCA, there is a decrease in the expression of AANAT and ASMT and the secretion of melatonin, changes that induce an increase in CCA growth by an autocrine mechanism; and 2) increases in the biliary expression of melatonin (by in vivo administration of melatonin or in vitro overexpression of AANAT in CCA cells) decreases CCA growth. MATERIALS AND METHODS Reagents were purchased from Sigma (St. Louis, MO), unless differently indicated. The antibodies against AANAT (FL-207), ASMT (C-20), MEL-1A-R (V-15), and MEL-1B-R (T-18) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA), unless differently indicated.

The rabbit polyclonal antibody against melatonin (ab35137) was purchased AV-951 from Abcam (Cambridge, MA). The ELISA kits for detecting melatonin levels in nonmalignant and CCA cell lines, and serum and bile from controls and CCA patients, were purchased from GenWay Biotech. In Vitro Studies Cell lines. We used six human CCA cell lines (Mz-ChA-1, HuH-28, TFK-1, CCLP1, SG231, and HUCC-T1) with different biliary origin. Mz-ChA-1 cells (from human gallbladder) (28) were a gift from Dr. G. Fitz (University of Texas Southwestern Medical Center, Dallas, TX).

01) and the cholesterol half-transporters Abcg5 and Abcg8 were decreased (P < 0.01 for both) (Table 2). ApoE overexpression reduced expression of the key enzyme for the alternative bile acid synthesis pathway in the liver, Cyp27a1 (P < 0.01) and the enzyme responsible for cholate synthesis, Cyp8b1 (P < therefore 0.05) but did not affect expression of the rate-limiting enzyme for classic bile acid synthesis, Cyp7a1 (Table 2). Hepatic gene expression of the sterol regulatory binding protein 2 (Srebp2) was suppressed by apoE overexpression (P < 0.05), whereas mRNA expression of its two target genes LDL receptor (Ldlr) and the rate-limiting enzyme for cholesterol synthesis, HMG-CoA reductase (Hmgcr), was not significantly affected (Table 2). There was also no difference in Abca1 mRNA levels in the liver between AdhApoE3 injected wild-type mice and controls.

In hCETP tg mice, virtually identical changes in liver gene expression were found in response to apoE overexpression (Supplementary Table III). Consistent with unaltered biliary secretion of cholesterol and bile acids in wild-type mice, the fecal mass output of neutral sterols (3.11 �� 0.22 vs. 2.49 �� 0.34 ��mol/day; Supplementary Table IV) and bile acids (2.33 �� 0.19 vs. 2.65 �� 0.11 ��mol/day; Supplementary Table V) was not influenced by overexpression of apoE. Likewise, hCETP tg mice injected with AdhApoE3 also excreted similar amounts of neutral sterols (Supplementary Table IV) and bile acids (Supplementary Table V) into the feces compared with AdNull administered controls.

Taken together, these results indicate that apoE overexpression promotes hepatic cholesterol uptake without increasing biliary and fecal sterol excretion. Hepatic apoE overexpression does not affect macrophage-to-feces RCT Because apoE overexpression increased hepatic selective uptake via SR-BI, an important step in the RCT pathway, but did not influence mass biliary and fecal sterol excretion, we next investigated whether apoE overexpression might affect overall RCT. In vivo RCT was traced after intraperitoneal injection of primary mouse macrophages loaded with 3H-cholesterol in control and apoE-overexpressing wild-type mice. The appearance of tracer in plasma was not significantly different between controls and mice injected with AdhApoE3 at 6 h (2.1 �� 0.6 vs. 3.0 �� 0.5% injected tracer dose; n.s.), 24 h (2.8 �� 0.4 vs. 2.

8 �� 0.3% injected tracer dose; n.s.), and 48 h (2.3 �� 0.2 vs. 2.4 �� 0.2% injected tracer dose; n.s.) after macrophage injection (Fig. 3A). Although apoE overexpression led to a 63% increase in macrophage-derived 3H-cholesterol within the liver (7.0 �� 0.9 vs. 11.3 �� 1.2% injected tracer dose; P < 0.05; Fig. 3B), overall, in vivo RCT remained essentially Anacetrapib unchanged as reflected by no effect on the total excretion of 3H-tracer into the feces (10.

Further inclusion criteria were an Eastern Cooperative Oncology Group performance status 0�C2 and adequate haematological (neutrophils 1.5 �� 109l?1 and platelets 100 �� 109l?1), renal (calculated creatinine clearance >50mlmin?1) and liver function (serum bilirubin 1.5 upper limit of normal range, liver transaminase or alkaline phosphatase selleck catalog concentrations 2.5 upper limit of normal range). No upper age limit was defined. Exclusion criteria were metastatic disease, previous chemotherapy for colorectal cancer or prior radiotherapy to the pelvis, history of another malignancy within the last 5 years, any contraindication to radiotherapy, clinically significant cardiac disease, malabsorption syndrome, peripheral neuropathy grade 1 according to National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTC version 3.

0), serious uncontrolled infection, concomitant treatment with any nucleoside analogue, known dihydropyrimidine dehydrogenase deficiency and psychiatric disorders or conditions interfering with compliance for oral drug intake. Pregnant or lactating woman were excluded. All patients provided written informed consent. The study protocol was approved by a local independent ethics committee and conducted in accordance with the Declaration of Helsinki. Pretreatment evaluation Before study entry, all patients were assessed by a multidisciplinary team comprising medical, radiation and surgical oncologists, gastroenterologists and radiologists. Patients underwent a medical history, physical examination, biopsy, ECG and staging studies (chest X-ray, abdominal�Cpelvis computed tomography scan, colonoscopy and endorectal ultrasound).

Pelvic MRI was optional but recommended for all patients with low-lying or T4 tumours. Complete laboratory tests included a full blood count, electrolytes, creatinine, liver transaminases, alkaline phosphatase, total bilirubin and carcinoembryonic antigen measurement. Treatment Radiotherapy Megavoltage equipment was used with 6�C18MV. Radiotherapy was delivered through three to four portal fields to the tumour and lymphonodal regions, and perirectal soft tissue structures at risk of microscopic disease. All patients received 45Gy, with a daily fraction of 1.8Gy given 5 days per week for 5 consecutive weeks. If treatment was interrupted, the dose was increased by 1�C2 fractions.

Chemotherapy Treatment consisted of a single cycle of XELOX (oral capecitabine 1000mgm?2 twice daily on days 1�C14 plus a 2-h intravenous infusion of oxaliplatin 130mgm?2 on day 1), followed by CAPOX combined with RT (capecitabine 825mgm?2 twice daily on days 22�C35 and 43�C56, and oxaliplatin 50mgm?2 on days 22, 29, 43 and 50; Figure 1). Chemotherapy and radiotherapy were interrupted if grade 3 or 4 toxicity was encountered (except for anaemia). Study Batimastat treatment was restarted when toxicity had resolved to grade 1. Dose reductions were required after grade 3�C4 toxicity.

sending the person described in the cholera vignette immediately to allopathic healthcare facilities, was considered as the least prominent category in the rural community, while it ranked fourth in the peri-urban community and was regarded as the most helpful thing one can selleck products do at home. The following statement from a housewife, aged 47 years, from Chumbuni is typical for what the communities would do for people with cholera at home: Table 5 Self treatment and help seeking for a cholera vignette in peri-urban and rural Zanzibar, n = 356 “At home we give water and other people give local treatment. [...] and if the condition becomes worse, we will send the patient to the hospital.” Public primary healthcare units and hospitals were mentioned by all respondents (Table (Table55 lower panel).

More than 95% of the peri-urban residents identified health facilities as most helpful source of treatment, while the rural residents’ preference was around 15% lower. Faith healers and traditional healers were of little importance and probing revealed that they would only be consulted after allopathic treatment had failed. Shigellosis versus cholera Similar to the cholera vignette, weakness was also rated as the most prominent somatic symptom for the shigellosis vignette (Table (Table66 top panel). Among symptoms related to dehydration, only loose skin and sunken eyes were mentioned; and both categories were reported significantly less for shigellosis than for cholera. The remaining symptoms of dehydration fell under the 5% threshold. All categories of somatic symptoms were differentiated on the individual level in both sites.

Table 6 Symptoms, perceived causes, and self treatment for a cholera and a shigellosis vignette in Zanzibar, n = 356 Notable among psycho-social problems was fear of infection and fear of isolation from others. Both categories were reported considerably less for shigellosis than for cholera, and were also well-differentiated (Table (Table66 second panel). All the other categories, which represent general features of diarrhoeal illness, i.e. costs, loss of family income, interference with social relationships and with daily activities, and being sad, anxious or worried, were not differentiated between both conditions (kappa coefficient greater than 0.4). A dirty environment was perceived to be the most prominent cause of shigellosis (Table (Table66 third panel).

The percentage of this category, however, was less than Drug_discovery half the percentage for cholera, and was closely followed by the category of eating unprotected or spoiled food. All categories of perceived causes that showed a significant difference between the two conditions were mentioned less frequently for shigellosis, with the exception of cannot say, which was reported three times more often for shigellosis than for cholera. Kappa coefficients for all categories were below the threshold of 0.4 suggesting differentiation of the meaning of cholera from shigellosis.

However, a defined guideline for a structured procedure of this technique was still lacking. The comparison selleck chemical Pazopanib of the visually controlled and performed pullback orientated at the electrode markers at the round window with the video-based measured amount of pullback showed a good relation of this distance. This finding indicates a good visual control of the procedure.An unintended tip movement was only found in three of the 3 �� 1 pullbacks, but not in 2 �� 1 or 3 �� 2 pullbacks. Therefore, a 3 �� 1 pullback cannot be recommended.The comparison of the optimum pullback distances showed an increase at number 1 and number 2 insertions of about 0.7mm between number 1 and number 2. This is almost the distance between the electrode markers. The difference between the number 2 and number 3 insertion is about 0.

35mm, supposedly due to an ��overinsertion��. Based on these data, the optimum pullback distance should be 1.37�C1.5mm (as for number 1 and number 2) in our view. The known variations of temporal bone size were taken into account in this study by initial radiological calculations of cochlear size. The lack of correlation between the size of the cochleae and the pullback could be based on the repetitive use of the electrodes as well as the known product-based interelectrode variability [15]. 5. ConclusionAn initial insertion to the first or second electrode marker followed by a pullback of 1.37mm to 1.5mm can be recommended. A pullback over the distance of two markers bears the risk of moving the electrode tip out of its initial position.AcknowledgmentThis work was supported by Cochlear Pty.

(Sydney, Australia).
Unmanned combat aerial vehicles (UAVs) are remotely piloted or self-piloted aircrafts that can carry many different types of accessories such as cameras, sensors, and communications equipment. They have a very wide range of applications that include both civil and military areas. Some important features that make them very popular are Brefeldin_A their low cost, smaller size, and their extended maneuver capability because of absence of a human pilot [1]. In particular, UCAV is one of the inevitable trends of the modern aerial weapon equipment, which develop in the direction of unmanned attendance and intelligence. Research on UCAV directly affects battle effectiveness of the air force and is a fundamental and significant research related to safeness of a nation. Trajectory generation and path planning is one of the key technologies in cooperative UCAV combatting.

The final demand vector, Y, can be treated as exogenous to the system; for example, the level of total production research use can be determined by the final demand (2):Y=BX.(2)Input-output model can be applied to calculate each sector’s indirect energy consumption regardless of the length and complexity of their production processes by using the energy input-output table (Wu and Chen, 1990; Peet, 1993). In energy input-output tables, energy sectors should be represented both in monetary and energy terms for computing the direct energy consumption coefficient matrix [14]. Assume that in input-output tables the economy can be categorized into n sectors, which includes k energy sectors and n-k nonenergy sectors.

Hence we can write an equation representing the way in which energy sectors distribute their products to energy sectors, nonenergy sectors, and final demands in physic units:Ak,1+Ak,2+?+Ak,k+Ak,k+1+?Ak,n+fk=xk.(3)Using energy input-output tables, the direct energy intensity and total energy intensity of each economic sector can be calculated. Direct energy intensity of one sector is calculated as the ratio of direct energy consumption (in physical terms) to total inputs (in monetary terms). Total energy intensities are calculated by multiplying direct energy intensity matrix with the Leontief inverse matrix of the corresponding energy input-output table. Embodied energy use in infrastructure investment can be calculated by multiplying total energy intensities with infrastructure investment:ei=��i=1kEj,1Xi,(4)etotal=e(I?A)?1,(5)EII=etotalYII=e(I?A)?1YII.

(6)ei is the direct energy intensity of sector i, e is the direct energy intensity matrix, and etotal is the total energy intensity matrix. EII is the embodied energy in infrastructure investment, and YII is the infrastructure investment.2.2. Structural Decomposition AnalysisBased on (6), the change of embodied energy use in infrastructure investment is driven by several factors, such as growth in infrastructure investment, energy efficiency AV-951 improvement, and industrial structure changes. Aiming at identifying the driving factors for changes in embodied energy use in infrastructure investment overtime, we applied input-output structural decomposition analysis on (6).

6, 2.2, 2.7, 3.2, and 3.8��L/cm2 of metabolites were sprayed in a cage (25cm length �� 15cm width �� 5cm depth) containing 25 mosquitoes. The exposed mosquitoes were kept under observation, and dead mosquitoes were discarded daily. Each bioassay including control was conducted in triplicate http://www.selleckchem.com/products/Calcitriol-(Rocaltrol).html on different days. In the control cages deionized water was sprayed. Daily mortality counts were performs. The bioassays were carried out at room temperature with 75 �� 5% relative humidity. The negative control was deionized water with 1% CDB while the positive control was Gokilaht-S 5EC (d,d-trans-cyphenothrin).2.4. Statistical AnalysisThe efficacy study of the filtrate metabolites of A. niger was assessed against Cx. quinquefasciatus, Ae. aegypti, and An. stephensi by probit analysis [17] with the statistical package IBM SPSS 19.

0.3. Results and DiscussionIn the present observations, we have evaluated the lethal effects of culture filtrates of A. niger against adult mosquitoes. The lethal effects of A. niger with LC50, LC90, and LC99 values of Cx. quinquefasciatus were 0.76, 3.06, and 4.751��L/cm2. Moreover, in case of the An. stephensi it was observed as 1.43, 3.2, and 3.86. While in case of Ae. aegypti it was recorded as 1.43, 2.2, and 4.1��L/cm2. These values were calculated after the exposure of seven hours along with their probit quotations (Table 1). The entomopathogenic fungus has been successfully reduceing mosquito vectors population in laboratories and field trials [2�C4, 18]. The fungal infections for the mosquitoes become increasingly sick and are eventually killed, but the process can take up to a week or more.

The adult mosquitoes pick up the fungal spores when resting on treated surfaces.Table 1Adulticidal activities of culture filtrates of Aspergillus niger against Culex quinquefasciatus (Say), Anopheles stephensi (Liston), and Aedes aegypti (Lin.). Unlike fast-acting chemical neurotoxins, fungal pathogens do not cause rapid mortality or immediate ��knockdown�� but rather act over a number of days as the fungal spores penetrate the insect cuticle and then proliferate within the hemocoel [19]. The A. clavatus has been found highly pathogenic against larvae of Ae. aegypti, Cx. quinquefasciatus, and An. gambiae [20]. The mortality rates were 100% against both Ae. aegypti, and Cx. quinquefasciatus, while against An. gambiae it was 95% after 24 hours.

The entomopathogenic fungus B. bassiana has used as an alternative vector control tool against insecticide-resistant mosquitoes under conditions typical of indoor resting environments [5]. A range of fungal-based insecticide combinations was used to test effects of timing and sequence of exposure. Both the laboratory-reared and field-collected mosquitoes were highly resistant Dacomitinib to permethrin but susceptible to B. bassiana and M. anisopliae infection, inducing 100% mortality within nine days.

Two parameters control the Introduction and Withdrawal procedure. The ToRecruitingProc parameter defines what part of population (from 0 to 1) will be subjected to the procedure. Another parameter, moreover WithdrAdd (from 0 to 1) specifies the probability that a given solution (W matrix) will be subjected to Gene Withdrawal or Gene Introduction. (If WithdrAdd = 0 then all solutions will go through Gene Introduction only; If WithdrAdd = 1 then all solutions will go through Gene Withdrawal only.)Figure 4Gene Introduction adds a new (n + 1) column (right-most) and row (lowest) to the W interaction matrix. Gene Withdrawal eliminates the right-most column and bottom row.2.3.2.

Involvement of the Recruited Gene in the Functioning of the GRN To quantify how much added genes affect network fitness, we used the following procedure: the model solution in a given generation was evaluated according to (2); then, for each additional gene (above the obligatory 2), fit to the data was evaluated with the W elements for that gene zeroed out. In cases where this produced a drop in fitness score (2) of more than 10% (a threshold determined in preliminary runs) compared to the full GRN, we kept the added genes as recruits to the GRN. We further filtered the most functionally significant recruits by use of a 33% threshold. Results are presented below for both threshold levels.2.3.3. Evaluation of GRN Robustness GRN solutions with E scores below threshold represent good fits to the gene expression data (Figure 3(a)). That is, these GRNs solve the problem of forming gap expression domains.

In addition to this, though, we want to test the robustness of these gap solutions to maternal variability. To do this, we took each good solution and tested its robustness to Bcd variability. We perturbed Bcd from Figure 3(b) according to[bcd]=[bcd]��[bcd]?Random?(0.2)?(3)(i.e., Brefeldin_A the Bcd profile varied within limits of ��20%). We reran the GRN with the perturbed Bcd values and compared these against the unperturbed result according toE��=��b(viaperturbed?viaunperturb)2.(4)This measure was calculated for 100Bcd perturbations for each GRN, and the results averaged for a measure of the GRN’s robustness (e.g., Figure 12).Figure 122-gradient model (Bcd-Cad) shows robustness to Bcd variability. Bcd��green; Kr��red; kni��blue. (a) Highly robust Kr and kni. (b) Nonrobust Kr and kni. (c) All borders are robust, except for anterior Kr. (d) Severe nonrobustness, …2.3.4. Artificial Transposons for GA The above Gene Introduction operator does not model the mechanism by which genes are incorporated into the genome.