A few years later, an epidemic of neurocysticercosis-related epilepsy was documented in that population.[2] On the other hand, the option that another food, that is, vegetables or fruits, infected with taenia eggs travel from one country to another

to infect patients has not been previously demonstrated and seems to be highly unlikely. Cysticercosis must be primarily GSK1120212 supplier considered as a disease transmitted from person to person, that is, from a taenia carrier.[3] Commenting on the other point raised by Joob and Wiwanitkit, what I meant by “reactivation of an infection that has previously been controlled by the host immune system” was that it may happen that the immune system of a given person infected Roxadustat chemical structure abroad, handled the infection without causing clinical manifestations. Then, several years later, the calcification that resulted from that successfully handled infection may become symptomatic when parasitic antigens (trapped within the calcified lesion) are liberated to the brain parenchyma and get exposed to the host immune system.[4] The resulting inflammatory response is responsible for the occurrence of seizures

(symptoms) and changes in neuroimaging studies that may resemble a fresh infection. Incorrect interpretation of imaging findings may lead the attending physician to believe that the patient has a cysticerci in the acute encephalitic phase (active neurocysticercosis).

“
“Background. Hajj, the pilgrimage to Mecca, is one of the obligatory religious duties of Islam. The travel clinic of the Public Health Service (PHS) Amsterdam administers vaccinations, including the required meningitis ACYW135 vaccine, and provides travelers with individual recommendations for all their travels. Methods. We extracted all data from the PHS database pertaining to Muslims who visited the clinic before travel to Mecca. From 2001 to 2009, Baricitinib the characteristics are described and trends are analyzed retrospectively. Acceptance of dTP vaccine was used as a proxy for acceptance of recommended vaccinations. For the years 2007 to 2009, predictive factors for the acceptance of advised vaccinations are analyzed. Results. From 2001 to 2009, significantly more women and people older than 50 years of age traveled to Mecca. Since 2007, only 527 of 2,156 (24%) of those who were advised to take vaccines accepted the recommendation. Independent factors for acceptance were being female, of younger age, and being less healthy. Specifically, Mecca travelers with heart disorders and with liver or gastrointestinal disorders accepted recommended vaccinations more often than those without. Conclusions. Only a quarter of Mecca travelers who visit the travel clinic for their mandatory meningitis vaccination also take other, recommended, vaccinations.

This suggests that in the absence

of other facilitators of transmission such as sexually transmitted infections, ART would be expected to be as effective in reducing infectiousness in men who have sex with men and other populations as it is in heterosexuals. Indirect evidence comes from a study of men who have sex with men attending HIV treatment services where ART was associated with a 96% reduction in HIV transmission [10]. Condoms should still be recommended to protect from other sexually transmitted infections, and to lower further any residual Ribociclib cost risk of transmission. Patients should be informed that taking ART does not result in immediate viral suppression. Studies have shown that the mean time to suppression of VL to <50 copies/mL in patients taking ART is about 90 days, and that a proportion may take 9 months or more [11]. Patients should also be informed about the possibility of virological failure leading to transmission of HIV. Decisions Smad3 signaling on condom use and safer sex should always be based on a recent VL test result and not on an assumption that taking ART implies non-infectiousness. For serodiscordant heterosexual couples wishing to conceive, irrespective of the method used for conception, the HIV-positive partner will need to be on ART with an undetectable plasma VL, regardless of his/her CD4 cell count or clinical status. This is likely

to reduce the risk of transmission sufficiently to be the only risk-reduction method some couples will want, but additional measures such as sperm washing, artificial insemination and potentially pre-exposure prophylaxis (PrEP) to the HIV-negative

partner have either been recommended in previous guidance [12] or are currently being assessed for couples wishing to address concerns of any residual risk of transmission. Details of the use of ART to prevent mother-to-child transmission are covered in the BHIVA guidelines for the management of HIV infection in pregnant women 2012 [13]. “
“The study aimed to assess the feasibility and acceptability of third-trimester antenatal HIV testing within our service after two cases of HIV seroconversion in pregnancy were noted in 2008. North American Guidelines recommend universal third-trimester HIV testing C1GALT1 in areas with an HIV prevalence of more than 1 per 1000. The HIV prevalence rate in our area is 3.01 per 1000. Pregnant women prior to 28 weeks of gestation were recruited at booking between 1 September 2008 and 31 August 2009 and offered an additional third-trimester HIV test. Consent was obtained and testing was performed by hospital and community midwives. Information was entered into a modified existing electronic maternity database. A qualitative e-mail survey of midwives investigated barriers to participation in the study. A total of 4134 women delivered; three (< 0.1%) declined first-trimester testing. Twenty-two women (0.5%) tested HIV positive, of whom six were newly diagnosed.

The questionnaire requested sociodemographic details, practice-related characteristics, and proposed three clinical situations with multiple choice questions (MCQ). To identify factors associated with a higher level of specific knowledge in travel medicine, results were studied by uni- and multivariate analyses. An overall score was calculated based on the MCQ answers and a motivation score was calculated based on parameters such as frequency and developments in pre-travel consulting at Rapamycin molecular weight the practice,

PCPs’ personal experience as travelers, and the formal agreement of PCPs to administer yellow fever vaccination. The response rate was 37.5%, with 150 questionnaires returned completed and suitable for analysis. After multivariate logistic regression, the three variables associated with a higher score were: proximity of a vaccination center (p = 0.001), motivation

score (p = 0.004), and absence of request for expert advice on malaria prophylaxis (p = 0.007). PCPs play an important role in travel medicine. This study showed that their high level of knowledge in travel medicine was mostly linked to their motivation to practice in this specialized discipline. Global international travel has increased considerably over the last few decades. The number of international travelers is roughly estimated at 900

million per year and should reach 1.6 billion per year in 2020.[1] Each BMS-354825 concentration year, 50 million people travel from industrialized countries to tropical areas. International travel from France mirrors this pattern, with around five million inhabitants visiting tropical areas each year.[1] Traveling abroad can lead to exposure to various diseases and following the expansion of international travel, primary care physicians (PCPs) are often consulted to provide medical pre-travel advice.[2] Travel next medicine is an emerging discipline born from the rising demand of the population but is not thoroughly studied by physicians. As the role of PCPs as first-line contacts for travelers seeking pre-travel advice has become increasingly significant, several worldwide surveys have investigated the quality of travel medicine practice among PCPs since 1987: four in the UK,[3-6] three in New Zealand,[7-9] two in Germany,[10, 11] one in America,[12] one in Qatar,[13] one in Australia,[14] and one in Switzerland.[11] In France, Bouldouyre et al.[15] recently published a survey focusing on the quality of pre-travel advice given by specialized physicians working in a travel medicine and vaccine center, but no study has yet focused on the quality of pre-travel advice given by French PCPs.

25% gastric mucin, 0.5% TA or 5% native PB and incubated at 37 °C for 24 h. Cells were harvested, washed twice with PBS, pelleted by centrifugation (3200 g × 15 min at 4 °C) and resuspended in PBS. CRB and SAT assays were performed as LY2109761 cost described above. Biofilm formation studies were performed using abiotic surfaces in sterile TPP flat-bottomed 96 well microtitre plates (MTP). Each well was filled with 200 μL of MRS broth supplemented with 0.25% gastric mucin, 0.5% TA, 5% PB or only MRS broth. A Lactobacillus

cell suspension (1.0 unit of OD620 nm= 1 × 108 cells) was added to each well and incubated under static conditions at 37 °C for 72 h. All plates were washed three times with sterile distilled water and bacteria attached to the surface were stained with 200 μL of 0.1% (w/v) CV in 1 : 1 : 18 of isopropanol-methanol- PBS solution (v/v) or

0.1% CR in PBS for 30 min (Kolter & Watnick, 1999; Nilsson et al., 2008). Excess dye was rinsed off by washing three times with water. The residual dye bound to the surface-adhered cells was extracted with 200 μL DMSO Vorinostat order and the OD of each well was measured at OD480 nm for CR or OD570 nm for CV in a MTP reader (Bio-Rad, Hercules, CA). To study early biofilm formation, 24-h-old biofilms grown in MTPs were washed twice with distilled water and fixed with 200 μL ethanol. Ethanol was allowed to evaporate by drying overnight at 37 °C and stained with CR and CV as described above. The amount of surface-bound CV or CR (in μg) was determined using a standard curve for the CR and CV, respectively. Values from all tests performed are the means of three separate experiments ± standard deviation. Statistical differences among the results obtained were

analyzed Thiamet G using one-way analysis of variance (anova) with minitab software (version 14.0; Minitab Inc., State College, PA). P values < 0.05 were considered significant. The comparisons made in the statistical analyses (one-way anova) are indicated in the figure legends. CRB of 17 lactobacilli strains was analyzed. Agar-cultured cells of auto-aggregating (AA) strains produced intense red colonies on CR-MRS agar, whereas broth-cultured cells developed weakly stained white colonies (Table 1). SAT and CRB of all strains grown on MRS agar and broth are shown in Fig. 1. In general, all strains except Lactobacillus rhamnosus and two L. gasseri strains showed high CRB when grown in agar MRS compared with strains grown in MRS broth. However, their SAT values were similar for agar- and broth-cultured cells (Fig. 1). A strong correlation was observed between the CRB and SAT results, with the three S-layer-producing L. crispatus AA strains, that is the most hydrophobic among all tested strains (Fig. 1a). Agar-cultured cells of L. reuteri DSM 20016 showed the highest CRB and lowest SAT values, whereas L. reuteri 17938 showed high CRB and a high SAT values (≥3.2 M). The CRB-positive curli-expressing E.

7,8 Spinal/vertebral TB, although rare among pregnant women, is associated with serious morbidity. Because PF-02341066 solubility dmso of non-specific symptoms, such as back pain (a common symptom in pregnancy), and reluctance to perform radiography in pregnant women, the diagnosis is often delayed, which can lead to early onset paraplegia.8,41 Paraplegia in pregnant women is associated with higher risk of urinary tract infection, decubitus ulcers, preterm labor, and autonomic hyperreflexia – a rare, but potentially fatal complication.41 Transportation of a paraplegic woman with TB is extremely difficult, as public transport in many developing countries is not patient-friendly, and private transportation is often not

affordable. Therefore, as a compromise, these women often skip antenatal care, what they need the most. Furthermore, surgical intervention for spinal TB during pregnancy is very challenging, and the expertise is limited to only a few super-specialty hospitals. Tuberculous kyphoscoliosis can also complicate maternal health and obstetric

management. Spinal deformity and reduced cardiopulmonary reserve associated with kyphoscoliosis can complicate the use of regional and general anesthesia, respectively, during delivery.42 In addition, surgical risk because of non-accessibility of lower segment can further complicate cesarean delivery. Extrapulmonary TB, especially tuberculous meningitis, is rare in pregnancy.11,43–45 Although it constitutes about 1% of all TB cases, only 55 LY2109761 order mafosfamide cases of tuberculous meningitis affecting pregnancy were identified up to 1999.43 Only a few cases have been reported from South Asian countries.11 However, an alarmingly high maternal mortality (38.2%), and fetal or neonatal deaths (36.6%) among these women remains a major concern.43 In our experience, misinterpretation of initial symptoms of meningitis with other infectious diseases can cause diagnostic delay resulting in dangerous complications, such as cranial nerve palsies and paraplegia. In these women, prolonged debility due to paraplegia and concurrent infections can also adversely affect the course of pregnancy

and perinatal outcome.11 Similarly, abdominal TB is exceedingly difficult to diagnose during pregnancy. Women may present with pyrexia of unknown origin. Tuberculous ascites, often masked by an enlarged uterus, rarely draws the attention of the clinician towards TB. Ascitic fluid studies (cytological, biochemical, and bacteriological) may provide evidence for TB, but with inordinate delay. Intestinal TB can present with subacute intestinal obstruction, and is mostly diagnosed by laparotomy.26 In certain cases, we find endoscopic biopsy or ultrasound-guided fine-needle aspiration biopsy to be very useful in pregnant women.8,26 However, such expertise is mostly limited to apex hospitals, and unfortunately, many women may not have access to such service on an urgent basis.

, 1995). The deletion mutant Δ19a was sensitive to menadione when grown anaerobically, which is not surprising considering that the ΔgrxAΔgsp E. coli double mutant was previously reported to be sensitive to H2O2 (Chiang et al., 2010). The deletion mutant Δ23a was the most sensitive to menadione when grown aerobically (Fig. 5) and lacked the barA gene,

which encodes a hybrid sensory histidine kinase in a two-component regulatory system with UvrY (Mukhopadhyay et al., 2000). BarA is involved in the transcriptional induction of RpoS. UvrY was already deleted in Δ17a (Pernestig et al., 2001). This study may ultimately allow the identification buy GDC-0199 of novel factors involved in the response to selleck compound oxidative stress. We found that the aegA gene was involved in menadione sensitivity and that the large-scale chromosome deletion mutant Δ1a lacking the aegA gene was menadione sensitive although a single deletion mutant of this gene was not menadione sensitive (Y. Iwadate & J. Kato, unpublished data). The deletion mutants may be useful for the investigation of alternate biochemical stress resistance pathways that might be cryptic in the wild-type strain. The deletion mutant with the most severely reduced genome was not the most sensitive to menadione under

aerobic or anaerobic culture conditions. Rather, menadione resistance tended to increase as additional deletions were combined in the same strain. The mechanism underlying this resistance is currently unknown but might involve the fine tuning of regulatory networks for defense against oxidative stress. Alternatively, the resistance might be related to the additional deletions, or to a point mutation or a spontaneous genome rearrangement that either might have occurred during the construction of the deletion mutants. These possibilities will

be investigated in a future study. A more detailed examination of the deletion mutants may reveal new genes involved in cryptic oxidative stress response pathways. We thank Y. Oguro, Y. Murakoshi, and M. Kobayashi for technical assistance. This work was supported by KAKENHI from the Ministry of Education, Culture, Sports, Science and Technology of Japan. Fig. S1. The DNA fragments used to construct the large-scale combined deletions. Fig. S2. Deleted chromosomal regions. Table S1. Deletion units and the primers used to construct them. Table S2. Sequences of the primers used to construct the deletion units. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Peptide deformylase (PDF) catalyses the removal of the N-formyl group from the nascent polypeptide during protein maturation.

97 Down-regulation of these proteins by hypoxia was associated with a decreased level of HRR activity as measured by the internal reporter system. Furthermore, they observed that decreased levels of

these HRR proteins was due to reduced translation of corresponding mRNAs, suggesting involvement of the mTORC1 or UPR pathways described above. They also demonstrated that depressing HRR by chronic hypoxia increased the cells’ sensitivity to the DNA cross-linking agents mitomycin C and cisplatin, and to radiation.92 Recently, a new pathway to down-regulate one of HRR gene, RAD52, by hypoxia was reported. Crosby et al. showed that HIF1-dependent up-regulation of the micro-RNAs, mir-210 and mir-373, results in suppression of RAD52 transcription.98 They showed that both micro-RNAs interact with the 3′ untranslated region of RAD52, suggesting that mir-210 and mir-373 are responsible for the repression Enzalutamide cost of RAD52.98 Taken together, these Birinapant results suggest that

depression of HRR by hypoxia may force a cell to use error-prone NHEJ that generates many genetic alterations. Yuan et al. showed that hypoxia increases the UV-induced mutation rate in tissue culture cells, suggesting that hypoxia represses nucleotide excision repair (NER).99 Later, Crosby et al. showed that hypoxia up-regulates mir-373, which in turn degrades the RAD23B transcript, one of the genes involved in NER.98 RAD23B recognizes UV-induced DNA damage in association with XPC and this complex recruits proteins, including XPA, RPA, XPB and XPD, for DNA unwinding. A small patch of single-stranded DNA containing damage is excised by XPG and XPF/ERCC1, and repaired and sealed by polymerase and ligase, respectively.100 Thus, repression of RAD23B by hypoxia can impair NER. During replication, DNA polymerase sometimes incorporates a wrong base generating a mismatch or generates a single-stranded loop within a highly repetitive sequence,

for instance at a microsatellite locus. These mistakes are repaired prior to mitosis by the MMR pathway. There are six MMR proteins involved in this system in humans, MSH2, MSH6, MSH3, MLH1, PMS2 and MLH3. The recognition of mismatch or loops containing one nucleotide Doxorubicin supplier is mainly mediated by MutSα (a heterodimer of MSH2 and MSH6). Recognition of a loop containing two or more nucleotides is mediated by MutSβ.101 Excision of a mismatched base or a loop on a newly synthesized strand is initiated by recruited MutLα (a heterodimer of MLH1 and PMS2) or MutLβ (a heterodimer of MLH1 and MLH3) and followed by exonuclease (EXO1). Re-synthesis is done by DNA polymerase and the nick is sealed by DNA ligase.101 If one of six MMR proteins is disabled, the mutation frequency in the microsatellite sequences increases. The microsatellite slippage mutations described above are associated with hypoxia-induced repression of MMR proteins, including MSH2, MSH6, MSH3, MLH1 and PMS2.85–87 Mihaylova et al. first demonstrated that severe chronic hypoxia (<0.

However, real false positives in industrialized countries are rare. Among 1000 amebic

serologies, Laverdant and colleagues reported only two cases of false positives concerning patients with hepatocarcinoma.[6] Thus, in industrialized countries, amebic serology must be performed only on patients with a hepatic abscess who have stayed in an endemic area. False negatives decrease sensitivity and negative predictive value. They can be due to patients’ immune response, the type of serologic test, or the pathogen strain. Sensitivity seems to be less important with PLX-4720 supplier serums obtained during acute illness (70%–80%) than those obtained during convalescence (>90%).[4] Indeed, a false-negative result can be obtained with a serologic test done within the first 7 to 10 days of the infection, but when repeated later, the test usually becomes positive: most of the time, seroconversion occurs before the 15th day. Furthermore, discrepant results can be seen between different assays done on the same sample. It has been described between LAT (negative) and IHAT (positive) in several publications (1/15 for Cummins[7] and 4/42 for Kraoul[8]), although these two assays use the same antigen. A similar result has been obtained between LAT and EIA (2/27 for van Doorn[9]). In this last case, initially negative samples

in LAT became positive 3 to 6 days later. Furthermore, E histolytica wild-type Cepharanthine strains compared to strains developed in cultures used to make serologic tests could present differences in their antigenic profile. selleck kinase inhibitor Tanyuksel and colleagues pointed out

that the lack of an accurately defined “gold standard” has impeded an objective assessment of the sensitivity of the antibody detection tests currently in use.[10] The accuracy of serology may be overestimated and the use of PCR methods tends to confirm this hypothesis.[11] Furthermore, no studies have been found concerning evaluation of amebic serology performance compared to a “gold standard” with likelihood-ratio test that limit the impact of prevalence. These observations lead to the conclusion that, in a highly evocative context with negative blood cultures, ALA should be considered despite a first negative amebic serology. Several propositions can be made to confirm the hypothesis of ALA. First, two or more screening serologic tests must be made. Second, if the initial serologic tests are negative, it is necessary to repeat the assay 7 to 10 days later. Third, the direct detection methods based on PCR gene amplification techniques realized in the abscess liver pus (when the aspiration is required) and also in blood, saliva, and urine samples appear to be very helpful and should be more systematically performed.[12] The authors state that they have no conflicts of interest to declare.

Forty-two (77.8%)

of 54 patients with AHC who received therapy started it before week 12. Further characteristics of both populations are listed VX-809 in Table 1. The IL-28B genotypes were in Hardy–Weinberg equilibrium (P=0.791 for AHC and 0.821 for CHC). The prevalence of the rs12979860 CC genotype was 47.5% among patients with AHC and 45.8% in those with CHC (P=0.778) (Table 1). In the group of individuals with AHC, 31 subjects with genotype CC (81.6%) were infected with HCV genotype 1 or 4 and 7 (18.4%) with genotype 2 or 3. Among CT/TT patients with AHC, 32 (76.2%) were infected with genotype 1 or 4 and 10 (23.8%) with genotype 2 or 3, respectively (P=0.948). In the group of patients with CHC, 119 (54.6%) of those with rs12979860 genotype CC were carriers of HCV genotype 1 or 4, while

99 (45.4%) were infected with HCV genotype 2 or 3. Of those harbouring rs12979860 genotype CT/TT, 200 (77.5%) bore HCV genotype 1 or 4 and 58 (22.5%) genotype 2 or 3 (P<0.001). A more detailed genotype distribution is shown in Table 2. In the subpopulation of patients with CHC enrolled in the German cohort, the distribution of HCV Z-VAD-FMK in vitro genotypes was also significantly different from that found in patients with AHC. Specifically, 41 (53.9%) German patients with CHC and rs12979860 CC harboured HCV 1 or 4 vs. 65 (75.6%) of those bearing CT/TT (P=0.034). There were no significant differences in HCV plasma load among patients with different IL-28B genotypes in the overall population. Thus, the median (Q1–Q3) HCV-RNA level was 6.36 (5.68–6.88) log10 IU/mL in carriers of rs12979860 CC and 6.27 (5.59–6.79) log10 IU/mL in those harbouring CT or TT (P=0.458). However, HCV-RNA load was significantly higher in patients with AHC and the CC genotype than of in those with AHC and rs12979860 CT/TT (Table 3). ALT levels in the entire population were higher in patients with the CC genotype [83 (58–165) in CC carriers vs. 74 (45–126) in CT/TT carriers; P=0.022]. The relationships between the IL-28B genotype and several parameters in the AHC and CHC groups are listed

in Table 3. Spontaneous clearance of HCV, as defined in this study, was documented in eight (10.1%) of the 79 patients in whom this information was available. There was no relationship between spontaneous clearance of the virus and HCV genotype. Thus, the numbers of patients who cleared HCV were as follows: six (11.3%) with genotype 1, one (12.5%) with genotype 2, one (10%) with genotype 3 and none with genotype 4 (P=0.746). The association between IL-28B genotype and spontaneous clearance did not reach statistical significance. Five (13.5%) of the patients with rs12979860 genotype CC and three (7.1%) of the patients with genotype CT or TT (P=0.349) showed spontaneous HCV clearance. The associations between IL-28B genotype and other factors are displayed in Table 3.

To confirm the gene deletion, transformants were screened by colony PCR. One of the

transformants yielded a product of about 1400 bp with the primer pair 1–1′, a product of about 1300 bp with the primer pair 2–2′ and a product of about 1700 bp with the primer pair 3–3′. PCR products of these sizes are expected in the case of a gene deletion of ku80 (see Materials and methods and Fig. 1). The Δku80 monokaryon was phenotypically indistinguishable from the wild type. The Δku80Δku80 dikaryon formed normal fruiting bodies that produced similar numbers of spores with a similar viability when compared Nintedanib supplier with the wild type. Moreover, like in the wild type, 109 protoplasts could be obtained from 5 g wet weight mycelium (data not shown). To assess whether the HR pathway was upregulated in the Δku80 mutant, qPCR was performed. Rad52 expression (which represents a gene involved in HR) was similar in the Δku80 strain (Ct=29.24±0.33) when compared with the wild-type strain (Ct=28.90±0.16). Apparently, inactivation of the NHEJ pathway does not induce an upregulation of the HR pathway. The Δku80 strain was transformed with the knockout constructs pDelcas-sc15, pDelcas-jmjC and pDelcas-priB. The deletion constructs had been linearized with the restriction enzyme SspI (pDelcas-JmjC and pDelcas-priB) or PacI (pDelcas-SC15) before they were introduced into the Δku80 H4-8 strain. Four out of seven

transformants had a deletion

of SB203580 datasheet sc15, while one out of one and two out of two transformants had a deletion of jmj3 and pri2, respectively (Table 2). Typically, 100 transformants are obtained with protoplasts of the wild-type strain and these transformants would contain one, if any, knockout strain (see the efficiency of the inactivation of ku80). The number of transformants obtained with the Δku80 strain is 100-fold lower (Table 2). However, most of these transformants have a gene deletion. Transformation of a Δku80 strain in which a wild-type ku80 gene had been reintroduced by crossing had a transformation frequency similar to that observed for the wild type. This confirms that the low transformation frequency was due to the deletion of the ku80 gene. The deletion of ku70 in Aspergillus oryzae (Takahashi et al., 2006) DNA Damage inhibitor and Sordaria macrospora (Pöggeler & Kück, 2006) also led to a reduction in the transformation frequency. In these cases, a seven- and 40-fold reduction in the number of transformants was obtained. Also in these cases, it may well be that the HR machinery is not upregulated when the NHEJ machinery is inactivated. The phenotype of the Δsc15 strain has been described (Lugones et al., 2004). Before determining the phenotypes of the Δjmj3 and Δpri2 strains, the wild-type ku80 gene was reintroduced by crossing. Monokaryotic and homozygous dikaryotic Δpri2 strains showed no phenotypic differences when compared with the wild type.