Both SOL and MP generated significantly higher amounts of IL-12p40 and IFN-γ

and lower amount of IL-10 showing a clear Th1 shift. Interestingly, 7 days after challenge, the IL-10 levels rebounded in the SOL group find more to levels comparable to that of Quadracel®. Thus, the MP formulation seems to maintain the Th1 response for a longer duration than SOL formulation. In summary, we demonstrated that immunization with PTd encapsulated into microparticles and adjuvanted with CpG ODN and IDR induced strong Th1 responses and partial protection against challenge with B. pertussis. From here on, future studies will determine whether inclusion of additional antigens like Pertactin and/or FHA in our formulations may result in enhanced protection comparable to commercial selleck acellular or cellular vaccines in a single shot model. This work was supported by a grant from the Bill and Melinda Gates Foundation through the Grand Challenges in Global Health Initiative and the Canadian Institutes of Health Research. Nelson Eng was supported by a post-doctoral fellowship from the Saskatchewan Health Research Foundation; Jason Kindrachuk received a fellowship from the Canadian Cystic Fibrosis Foundation; REWH holds a Canada Research Chair in Microbiology. We acknowledge Jill van Kessel,

Stacy Strom, Rachelle Buchanan and the Animal Care personnel at the Vaccine and Infectious Disease Organization for their assistance in this project. This manuscript has been approved by the Director of VIDO as manuscript#582. “
“In the course of replication most viruses make defective-interfering (DI) viruses, which are virus particles composed of a normal set of viral proteins encapsidating a deleted version of the viral genome. Because they lack essential genetic information, DI

viruses are replication deficient. Replication of the defective genome is achieved by the presence in the same cell CYTH4 of a genetically compatible infectious genome, usually from the virus that generated the DI genome, and which provides the missing function(s) in trans. DI virus is thus totally dependent on infectious virus for replication. Interference occurs when the ratio of defective: infectious genomes increases to a level which results in a reduction of the amount of infectious virus produced [1], [2], [3], [4] and [5]. Most of our knowledge comes from studies in cultured cells, but there is also limited evidence that DI virus can protect against virus diseases in vivo [6], [7], [8], [9] and [10]. The conventional view, developed by extrapolation from in vitro studies, is that the protection afforded in vivo is also due to competition between the DI and infectious viruses at the level of genome replication. However, in those cases where in vivo protection has been seen there is little direct evidence for this or any other mechanism.

Unit costs for the treatment of CIN2/3 in each country are shown in Table 2. Costs were expressed in local currency and updated to 2011 value using the country-specific Consumer Price Index reported by the World Bank for each country [20]. Fig. 1 presents country level results grouped by WHO continent

and worldwide of the estimated annual numbers of CC cases potentially avoided by HPV vaccination at steady-state at varying levels of vaccination coverage. Individual country estimates at four levels of vaccination coverage (50, 70, 90 and 100%) are shown in Supplementary File 1. In all five WHO continents, numbers of cases potentially Selleckchem Abiraterone prevented by vaccination was at least 18% greater in the analyses including cases causally related to HPV irrespective of type, compared with the cases causally related to HPV-16/18 infection only. The relative difference (i.e. the percentage increase of cases avoided causally related

SB203580 to all HPV types vs. HPV-16/18 only) was most pronounced in Africa (34%). Relative increase of number of cases avoided for other WHO continents was 27% for America, 26% for Asia, 21% for Europe, 18% for Oceania and 27% worldwide. A similar pattern was observed for the estimated annual numbers of CC deaths potentially prevented by HPV vaccination (Fig. 2). Similarly to CC cases prevented, the inclusion of CC deaths prevented irrespective Megestrol Acetate of HPV type in the analysis increased by at least 18% the estimated number of deaths potentially avoided, with the relative difference having the same values as for CC cases analysis. Individual country estimates

for the CC deaths potentially prevented at four levels of vaccination coverage (50, 70, 90 and 100%) are shown in Supplementary File 2. Table 3 shows the estimated annual cost-offset associated with CC prevention at steady-state in Mexico, Canada, Germany, Thailand and South African Republic. Including VE irrespective of HPV type in the analysis increased the estimated cost-offset in all five countries by at least 10 million Int$. Table 4 presents the estimated annual numbers of CIN2/3 cases avoided by HPV vaccination at steady-state in Italy and Malaysia. The estimated vaccine impact on CIN2/3 cases, and treatment costs averted were 33 and 53% higher in Italy and Malaysia respectively, for the analysis irrespective of HPV type, compared with the estimates for HPV-16/18 only. The results presented here suggest that HPV vaccination of young girls naïve to HPV with the AS04-adjuvanted HPV-16/18 vaccine could reduce the number of CC cases and deaths in countries worldwide, with the absolute number of CC cases and deaths and hence, lives saved depending on the vaccination coverage achieved.

Data from the activity monitor were deidentified at AZD4547 downloading to allow assessor blinding for average and total energy expenditure. The participants’ perception of using a gaming console as an exercise modality was measured using a 10-cm horizontal visual analogue scale. Participants were asked to rate their level of enjoyment, fatigue experienced, and workload achieved during the exercise intervention. In addition, participants were asked to rate their

confidence that the exercise intervention met their perception of an effective exercise for them and that the exercise intervention was feasible to be included as a component of their routine exercise regimen. All visual analogue scales were anchored, with the left hand anchor indicating no agreement with the statement (no enjoyment, not fatiguing, no workload, not effective, not feasible) and the right hand anchor indicating strong agreement (very enjoyable, very fatiguing, etc). Cardiovascular demand and energy expenditure measures were recorded continuously during 5 minutes of rest PD98059 in vivo at the start of the exercise interventions and during the 15 minutes of exercise. The participants’ perceptions of

the exercise intervention were measured at the completion of the exercise. The primary outcome was the average heart rate during exercise. We planned and undertook an analysis of the first 14 participants to determine the standard deviation of the difference between two recordings of the average heart rate during exercise in the same patient, which was 12 beats/min. In the absence of an established value, we nominated 10 beats/min as a clinically worthwhile difference in heart rate during exercise based on our clinical experience and because it exceeds day-to-day variability in heart rate (Achten and Jeukendrup 2003).

Therefore, a sample size of 18 participants was required old to achieve 90% power to detect a difference of 10 beats/min between the two exercise interventions at a significance level of 0.05. All measures were analysed using an intention-to-treat analysis. Means and standard deviations were calculated for all variables. Average, minimum and maximum values were recorded for heart rate and oxygen saturation during the 5-minute rest period and the 15-minute exercise period for each exercise intervention. Average energy expenditure during the 15 minutes of exercise was estimated by the activity monitor software in metabolic equivalents (MET). Total energy expenditure for the entire exercise intervention was estimated in kilocalories by the same software. Differences in all variables between the two exercise interventions were analysed using paired t–tests. Results were reported as mean differences and 95% CI. Statistical significance was set at 0.05.

8%) had glaucoma in both eyes. Seventeen of all included patients (2.9%) were registered in the administration system of the Habilitation and Assistive Technology Service

only. Median time between last visit and death was 8 months learn more (interquartile range 3-16 months). Median age at death was 87 years (range 50-103 years). There were 423 patients in the Data at Diagnosis group (71.5%). In those patients mean age at diagnosis was 74.0 ± 7.9 years, ranging from 46-95 years. Exfoliative glaucoma was found in at least 1 eye in 170 patients (40.2%). Average perimetric MD at diagnosis was −5.59 ± 5.69 dB and −11.83 ± 8.18 dB in the better and the worse eye, respectively. Median VA at time of diagnosis was 0.8 (20/25), ranging from no light perception to 1.00 (20/20), in the perimetrically better eye and 0.8 (20/25), ranging from no light perception to 1.25 (20/16), in the perimetrically selleck compound worse eye. Untreated mean intraocular pressure (IOP) value in all glaucomatous eyes at time of diagnosis was 27.2 ± 8.8 mm Hg. Numbers of patients with low vision and blindness from glaucoma at the last visit are shown in the Table. At the last visit, 42.2% (250 of 592 patients) of all patients were blind from glaucoma in at least 1 eye and 16.4% in both eyes. Other reasons for unilateral blindness

were age-related macular degeneration (AMD) (26 patients), a combination of cataract and other disease (10 patients), and other causes (32 patients). Seventeen patients were bilaterally blind because of reasons other than glaucoma (16 from AMD, 1 patient from other reason). A

combination of causes for blindness was found in 1 eye of 7 blind patients (Table). There was no statistically significant difference in the frequencies Ketanserin of visual impairment at the last visit when comparing the Data at Diagnosis group and the Follow-up Only group (Table, P = .260). In patients who developed blindness attributable to glaucoma, the median time with bilateral blindness was 2 years (<1-13) (mean 3.0 ± 3.1). Patients who became bilaterally blind from glaucoma did so at a median age of 86 years (range 66-98; mean 85.7 ± 6.1). Only 13 patients (13.5% of blind patients and 2.2% of all patients) became blind before the age of 80 years. The median duration with diagnosed glaucoma was 12 years (<1-29) (mean 11.2 ± 6.6), and 74.7% (316 of 423 patients) of patients had their glaucoma diagnosis for more than 6 years. The cumulative incidence for blindness in at least 1 eye and bilateral blindness from glaucoma was 26.5% and 5.5%, respectively, at 10 years and 38.1% and 13.5%, respectively, at 20 years after diagnosis (Figure 3, Top left and Bottom left). The corresponding cumulative incidences for blindness caused by other reason were 0.7% and 0.7%, respectively, at 10 years and 2.4% and 2.6%, respectively, at 20 years (Figure 3, Top left and Bottom left). The Kaplan-Meier estimates for blindness in at least 1 eye caused by glaucoma were 33.1% at 10 years and 73.

Subgroup analyses stratified by age group, performance status, histology/tumor grade, or stage/debulking status were also conducted. A total of 462 patients were enrolled in this study, with 276 evaluable for inclusion in the analysis (Figure 1). Patient characteristics are displayed in Table 1. The median age of

the study population was 61 years, and most patients had tumors that were classified as papillary serous (84%), poorly differentiated (83%), stage III (85%), and optimally debulked (72%) (Table 1). The majority (94%) completed 4-8 cycles of chemotherapy. The median follow-up period was 23 months (range, 12–37 months), and 193 (70%) patients experienced this website disease progression within this time frame. The median PFS was estimated to be 15.9 months (95% confidence interval [CI], 14.3–17.1 months). Assay results for carboplatin were available for 231 patients, with 44 (19.1%) patients identified as resistant to this therapy in the chemoresponse assay. Assay data for paclitaxel were available for 226 patients, 49 (21.7%) of whom were classified as resistant. Assay resistance by age, performance status, histology/grade,

and stage/debulking status is summarized in Table 2. There is no evidence that assay result for either carboplatin or paclitaxel is correlated with patient characteristics. Assay result for carboplatin was significantly associated with clinical outcome (Figure 2). The median PFS was 16.6 and 11.8 months for assay nonresistant (sensitive + IS) and resistant tumors, respectively. Patients displaying assay resistance to MI-773 datasheet carboplatin were at a higher risk of disease progression as compared to those who were nonresistant (HR, 1.87; 95% CI, 1.29–2.70; P < .001). These results were Rebamipide consistent in multivariate analysis after controlling for clinical covariates (HR, 1.71; 95% CI, 1.12–2.62; P = .013) ( Table 3). Analysis of subgroups (age group, performance status, histology, stage/debulking status) was also conducted ( Figure 3), and the association between PFS and assay result for carboplatin was suggested across all subgroups.

The data also suggest that patients with assay resistance to paclitaxel would experience shortened PFS, but the association did not reach the level of statistical significance ( Table 3). Assay results for carboplatin and paclitaxel were highly correlated. For 220 patients with assay data available for both agents, 75.5% were nonresistant to both agents and 15.9% were resistant to both agents, while only 8.6% of patients were resistant to only 1 agent (5.9% to carboplatin and 2.7% to paclitaxel). Patients resistant to both agents experienced the worst outcomes (HR, 1.66; 95% CI, 1.10–2.52; P = .017, as compared to patients nonresistant to both agents). Multivariate analysis indicated the same tendency, although the association was not statistically significant ( Table 3).

NS-EA 51 and Famotidine caused high significant (P < 0.001) reductions in ulcer index. However fraction did not alter significantly, gastric wall mucus content in hypothermic-restrain stressed gastric ulcer model rats while Famotidine significantly (P < 0.05) inhibited this effect in the treated animals ( Table 2). The anti-ulcer action of N. sativa seed powder (NS) its ethanol extract (NS-E), ethyl acetate fraction (NS-EA) and purified fraction Epacadostat purchase (NS-EA

51) viz., inhibition of gastric aggressive factors (acid and pepsin), due to the ability to interfere with the indomethacin induced-inflammatory and PGE2 synthesis inhibitory effects, reported earlier. 9 Lipid peroxidation and anti-inflammatory activities of various constituent/extract of N. sativa showed by Suboh et al. 20 and Hajhashemi et al. 21 respectively see more were found in accord to our

findings. In the present study, the anti-ulcer action of NS-EA 51 was further evaluated in the histamine plus PL and hypothermic-restrain stressed rat models. It has been reported that histamine plays important role in causation of inflammation, allergy, gastric acid secretion, neurotransmission, embryogenesis and in development of various tumors.22 In gastric parietal cells, three types of receptors such as histaminic H2-receptors, muscarinic receptors (M1) and gastrin receptors (G) have been reported. Out of these, histamine receptors have been found to play major role in gastric acid secretion. Histamine-enhanced gastric acid secretion along with acid-output oxyclozanide has been reported to reach the maximum level and plateau immediately

and 1.0 h after of histamine administration.3 and 23 Acid stimulation in the stomach has been reported to be mediated by histamine, released from the mucosal mast cell, which has been indicated one of the cause of mucosal damage (ulcer formation).11 and 14 Moreover, among various tools used to provoke gastric ulceration in animal models, restraint plus cold water-immersion has been reported to act synergistically and gives reproducible and reliable results.24 Cold-restraint stress-induced gastric ulceration and the possible mechanisms have been found to involve an increase in the inhibitory γ-aminobutyric acid (GABA) and suppression of stimulatory nor-epinephrine (NE) and dopamine (D) in central regions, especially the cerebral cortex and/or thalamus/hypothalamus.25 Vagal stimulation (stimulation of the hypothalamus, directly or indirectly) has been thought to be one of the mechanism for increase in gastric acid secretion. The mechanism of experimental stress-induced ulcers has been found to be dependent on an interaction between the presence of acid, changes in mucosal circulation, an increase in excretion of glycoproteins in mucus and a decrease in mitotic activity of the mucosal lining of the stomach.24 Moreover, endogenous PGI2 has been found to be involved in the gastric ulcerogenic response to stress.

4 years for the bivalent vaccine with 100% seropositivity maintained and at least 5 years for the quadrivalent vaccine with 98.8% seropositivity see more maintained

[24]. The bivalent vaccine induces sustained antibody titres for HPV18 several fold higher than after natural infection, 8.4 years after initial vaccination with 100% seropositivity maintained. However, for the quadrivalent vaccine, 18 months after first vaccination, the induced antibody titres for HPV18 return to the level of natural infection, with a reduction in seropositivity over time [42]. A correlate for protection has not yet been established and further studies will determine whether these decreasing antibody levels are linked to reduced effectiveness. The immunogenicity of the bivalent and quadrivalent vaccine was Nutlin-3a nmr compared in a head-to-head trial. Neutralising antibodies (nAbs) against HPV16 and HPV18 were 3.7 and 7.3-fold higher, respectively for the bivalent vaccine compared to the quadrivalent vaccine in women of age 18–26 years old at month 7 after receiving the first dose [43]. These differences remained similar in older age groups. After 24 months of follow-up, the GMTs of nAbs were 2.4–5.8-fold higher for HPV16 and 7.7–9.4-fold higher for HPV-18 with the bivalent versus the quadrivalent vaccine [24] and [44]. This observation remained similar up to 48 months of follow-up: GMTs of nAbs were consistently

higher in those receiving the bivalent vaccine across all age strata: 2.0–5.2-fold higher for HPV16 and 8.6–12.8-fold higher for HPV18 [45]. The use of different adjuvants in the vaccines might explain these differences in immunogenicity [46]. The difference in immune response observed at month 7 between the two vaccines was sustained up to month 48. However, the long-term clinical implications of these

observed differences in antibody response need to be determined. An anamnestic response was observed after the administration of a fourth dose after 5 years for the quadrivalent vaccine [47] and after 7 years for the bivalent vaccine [48]. In a phase I/II study in South Africa, the bivalent HPV vaccine was shown to Rolziracetam be immunogenic and well tolerated in HIV-infected women up to 12 months after vaccination. All subjects, both HIV-positive and HIV-negative were seropositive at month 2, 7 and 12, although antibody titers were lower in HIV-positive children [49]. Similar results were observed with the quadrivalent vaccine [50]. Several studies are currently on-going in HIV-positive adolescent girls and young women to evaluate the safety and immunogenicity of HPV vaccines [17]. Both HPV vaccines have some cross-protection against types that are not included in the vaccines, possibly explained by phylogenetic similarities between L1 genes from vaccine and non-vaccine types: HPV16 is phylogenetically related to HPV types 31, 33, 52 and 58 (A9 species); and HPV18 is related to HPV45 (A7 species).

Paper discs with test compounds were placed on agar surface at proper distance. The plates with test compound discs were incubated at 37 °C for 24 h. The minimum inhibitory concentration (MIC) of each

compound was determined by observing the zone of inhibition around each disc. The title compounds are screened for antibacterial Selleckchem Ibrutinib activity by employing paper disc method. The bacterial strains used are S. aureus (Gram +ve) and E. coli (Gram −ve). The substituted 4,5-dihydro-4-oxothieno[3, 2-c]quinolines revealed considerably good antibacterial activity against the bacterial strains. Of them, 3-amino-4,5-dihydro-5-ethyl-4-oxothieno[3,2-c]quinoline-2-carboxylic acid (2d) exhibited most promising antibacterial activity against S. aureus at 4 μg/disc concentration and against E. coli at 200 μg/disc concentration. Antibacterial activity associated with the title compounds was evaluated by comparing with the standard antibiotic drug, ciprofloxacin, which is active on Gram +ve and Gram −ve bacteria. Surprisingly many of these novel heterocyclic

compounds exhibited potent antibacterial activity against S. aureus (Gram +ve), but did not show any activity against E. coli (Gram −ve) even at 200 μg/disc concentration. Solvents employed in the present investigation were tested for antibacterial activity and found Afatinib supplier to be inactive on both the bacteria. Many crystal structures are available in PDB for S. aureus DNA Gyrase (PDB IDs – 2XCO, 2XCQ, 2XCR, 2XCS, 2XCT), one of which has Ciprofloxacin as the co-crystallized ligand (2XCT) with a resolution of 3.35 Å. We considered a high resolution Rolziracetam (2.1 Å) crystal structure of S. aureus DNA Gyrase for our studies, but the active site data was taken from 2XCT (Ciprofloxacin binding site). The residue Ser1084 was found to be the key residue of the active site which makes a hydrogen bond with Ciprofloxacin. The protein was prepared for docking study using the Protein Preparation Wizard of Maestro. Water molecules were removed, Hydrogen were added and the protein was minimized (only Hydrogen) using OPLS 2001 force field ( Fig. 2). The synthesized compounds were constructed and prepared for docking using the Ligprep

Protocol of Maestro. Ligand minimization was done using OPLS 2005 Force field. The minimized protein and ligands were uploaded to GOLD 3.2 for docking. The active site radius was set to 10 Å form the atom number 4158, the oxygen atom of the active site residue Ser1084, which forms hydrogen bond with Ciprofloxacin. All the default values for annealing parameters (van der Waals = 4.0, H-Bonding = 2.5) and Genetic Algorithm Parameters (Population Size = 100, Selection Pressure = 1.1, No. of operations = 10,000, No. of Islands = 5, Niche Size = 2, Migrate = 10, Mutate = 95, Crossover = 95) of GOLD were used for docking (Fig. 2). Four title compounds (Fig. 1, 1a–d) were tested and the results are included in Table 2. All of them were active against S.

At an increased frequency of measles outbreaks, such a diversion of public health resources to

outbreaks response could significantly consume public health budgets, divert the health priorities and roles at the local and state levels and further increase the pressure on available resources. As an illustration of the opportunity costs imposed on public health departments, we estimated that the number of personnel hours responding to these sixteen measles outbreaks would require the full time work of 20–39 public health officers during a year (i.e., assuming 2080 h/year or 40 h/week). Likewise, including cost of other inputs and materials, each public health department that Perifosine nmr experienced a measles outbreak in 2011 would have incurred a median range cost of $11,933–$29,833 per measles case. These costs, however, are not exclusive of measles outbreaks since about 113 (51% of the 220) measles

cases reported in 2011 occurred by definition not in outbreak settings yet they may have demanded a similarly resource-intensive response from local public health departments. A very conservative estimate (i.e., assuming only three contacts per case) of the impact of the 113 non-outbreak RO4929097 concentration measles cases – isolated or fewer than three epidemiologically linked cases – would add approximately 1579 personnel hours and would increase total costs by approximately $100,128. Measles outbreaks will likely continue to occur in the US mainly because of the persistent risk of imported measles cases derived partly from the increased disease transmission and number of outbreaks in the European

region [21]. Such a risk is magnified by the presence of susceptible sub-populations in the US due to lack of vaccination, the variety of potential outbreak settings (hospitals, clinics, airports, cruise ships, etc.), the limited state and local response capabilities, and the lack of awareness of vaccine recommendations in a few others susceptible individuals traveling to endemic countries. Beyond the impact on local and state public health departments, responses to measles outbreaks also affect hospitals, clinics [9] and [22], as well as non-health public departments such as schools, universities and occasionally local police departments enforcing quarantines or supporting control actions [11] and [13]. Additionally, susceptible individuals and their households face higher health risks derived from potential serious measles complications (i.e., otitis media, pneumonia, encephalitis or death [23]) along with associated medical and productivity lost costs [23] and [24]. This study has some limitations. The personnel costs used for this study were based on average estimates of data reported in four previous studies published before 2011.

For stabilization of SLNs, the surfactant forms a coating layer so that lipid nanoparticles do not coalesce.5 The second-order polynomial equation relating the response

of % entrapment efficiency (Y2) is given below: equation(2) Y2=+67.81+2.84A−0.71B−3.39C−0.78AB+0.69AC−1.36BC+1.74A2−4.06B2+0.22C2Y2=+67.81+2.84A−0.71B−3.39C−0.78AB+0.69AC−1.36BC+1.74A2−4.06B2+0.22C2 The model F-value of 69.33 implied that the model is significant (p < 0.0001). The ‘Lack of Fit F-value’ of 0.099 implied that the Lack of Fit is not significant (p = 0.9563). As Table 3 shows, the ANOVA test indicates that A, B, C, AB, BC, A2 and B2 are significant model terms. Positive coefficients of A, AC, A2& C2 in equation (2) indicate the synergistic effect on % entrapment efficiency, while negative coefficients of B, C, AB, BC, & B2 indicate the antagonistic effect on % entrapment efficiency. The “Pred R Squared” of 0.9716 is in reasonable agreement Palbociclib price with the “”Adj R-Squared”" of 0.9746, indicating the adequacy of the model to predict the response of entrapment efficiency. The ‘Adeq Precision’ of 34.30 indicated an adequate signal. Therefore, this model is used to navigate the design space. The 3-D surface plots for % entrapment efficiency are shown in Fig. 2. The effect of drug to lipid ratio on %

entrapment efficiency depends on the extent of drug solubility in lipid. An increase in % entrapment efficiency from 62.76 (H1) to 69.87 (H2) was observed on increasing the drug lipid ratio from 1:2 to 1:4 (Table 2). This is due to large amount of lipid present for drug entrapment. On further increasing drug to lipid IOX1 datasheet ratio the entrapment efficiency decreased

(data not shown). This is due to expulsion of drug from particle surface.11 A decrease in % entrapment efficiency from 69.00 (H13) to 65.32 (H12) was observed on increasing surfactant concentration and stirring speed (Table 2). The probable mechanism of this behaviour could be that as the particle size decrease on increasing stirring speed, the surface area increase. As the surfactant increase at a constant amount of lipid, the surface of the formed SLNs is too small to adsorb all surfactant molecules, which will about result in the formation of micellar solution of the drug. Hence, the solubility of the drug in water phase will be increased. Therefore, the drug could partition from SLNs into the formed micelles in the water phase during stirring or washing time.12 The second-order polynomial equation relating the response of % drug loading (Y3) is given below: equation(3) Y3=+18.43−4.83A−0.16B+0.68C−0.14AB−0.21AC−0.34BC+1.6A2−0.81B2−0.019C2Y3=+18.43−4.83A−0.16B+0.68C−0.14AB−0.21AC−0.34BC+1.6A2−0.81B2−0.019C2 The model F-value of 323.46 implied that the model is significant (p < 0.0001). The ‘Lack of Fit F-value ‘of 3.64 implied that the Lack of Fit is not significant (p = 0.1221).