Symptoms may be discreet and comprise an urticarial rash and/or a

Symptoms may be discreet and comprise an urticarial rash and/or angio-edema, medium grade fever, a non-productive cough, abdominal pain, and diarrhea.5,10,19–22 In most patients of this cluster, symptoms were mild and had already resolved before treatment was given. In practice, AS is usually not recognized by primary

health care providers who are not familiar with tropical pathology. When the first symptoms appear, eosinophilia may not yet be raised.10 As illustrated with this cluster, eosinophilia will however increase rapidly in the course of the following days to levels rarely seen in other parasitic PTC124 diseases. Diagnosis is thus likely when at least one of the above symptoms appears in association with a clearly raised eosinophil count and with a primary exposure to schistosomiasis up to 90 days prior, pending confirmation of schistosome infection.1,23,24 In the early disease stage, diagnosis cannot be reliably confirmed by antibody tests or parasite detection methods.6,25 However, by the time patients are referred to a travel clinic, evidence of schistosomiasis

is found in most, mainly by serum antibody detection and/or ova detection in feces or urine.6,10,23,25 The current techniques for the laboratory diagnosis of AS have some shortcomings. Antibody production against adult worm and egg antigens starts only after schistosomules in the liver have been matured and after oviposition has started around the perirectal or perivesical venous plexus. This occurs at the earliest FDA approved Drug Library 6 weeks after infection, when symptoms may have largely subsided. Serological techniques used in clinical practice do not distinguish active infection from past exposure nor provide reliable information on parasite burden, and are not species-specific. Most routine techniques detect IgG, IgM, or IgE against soluble worm antigen (SWA) or soluble egg antigen (SEA) by ELISA, HAI or immunofluorescence. When combining assays using different sets of antigens in parasitologically confirmed infection, sensitivity may exceed 90% while retaining specificity at over

97%, but is less in AS.10,11,26,27 In this cluster, seroconversion failed to happen in three patients during the follow-up period, one parasitologically confirmed and two with symptomatic AS. Whether this is due to early treatment, a low parasite burden, or host Cyclic nucleotide phosphodiesterase immune response factors is unclear. In most travelers and migrants, established schistosomiasis infection is predominantly asymptomatic and with a low parasite burden, so that eggs are often not found in excreta.28,29 Nevertheless schistosome eggs were detected in feces of nearly all (6/7) symptomatic patients of this cluster. This may be due to low average age of patients, as well as to the relatively long average time lapse between exposure and diagnosis.30 There is thus a need for a more sensitive qualitative diagnostic test that confirms schistosomiasis infection at an earlier stage.

It is sometimes difficult to decide if one foot is warmer than no

It is sometimes difficult to decide if one foot is warmer than normal (e.g. due to infection or Charcot foot) or, if in fact, the other foot is cooler due to PAD. Redness of the foot may occur in infection, but is also seen in severe PAD (Figure 1). PAD may also mask the inflammatory response to infection so learn more the

signs of infection may be very subtle or missed. Infection can also lead to discomfort or pain in the ischaemic foot and can be the trigger for the development of CLI in an ‘at risk’ foot. Palpation of the foot pulses includes the presence or absence of the posterior tibial, and dorsalis pedis pulses (up to 10% of the normal population do not have a palpable dorsalis pedis). It is exceedingly unusual to have a clearly palpable foot pulse in advanced CLI. The main exception to this would be distal small vessel embolisation causing localised tissue infarction. When there is uncertainty about the presence of a pulse it is best to assume that the pulse(s) is

absent and arrange further investigation. Assessment for any lower limb neuropathy is also vital.3,20 All people with diabetes should undergo annual foot screening, including palpation of foot pulses3,20 by a suitably trained health care professional,4 with subsequent classification of their current risk status, and a management plan then agreed with the patient. If found to be other than at low current risk (i.e. increased/moderate or high risk), without current active foot disease,

then they should receive review by a member of the ‘foot protection team’3,4 or a podiatrist20 at regular intervals.3,20–22 Although, as mentioned above, the diagnosis of CLI is highly unlikely in the presence of unless a clearly palpable foot pulse, the presence of a foot pulse does not exclude the diagnosis of PAD. ABPI may be useful in this situation as a supporting diagnostic test. Of course, all active foot disease, e.g. new (or deteriorating) foot ulcer, discolouration, swelling, or CLI (with or without tissue loss) should be referred rapidly (within 24 hours) to the specialist diabetes ‘multidisciplinary foot team’ (MDFT).3,20,22,23 Although further investigation is possible outside specialist centres, e.g. ABPI (see below), if CLI is suspected on the grounds of a simple but thorough history and examination, then urgent onward referral is indicated. For patients with diabetes and associated tissue loss or ulceration then this would usually be to the specialist diabetes MDFT. Where pain is the predominant symptom, without tissue loss, this may be to the vascular team depending on local pathways. No matter what the local pathway, it is vital that urgent referral and subsequent review are arranged.

Each CS was presented for 10 s during the experiment and the US w

Each CS was presented for 10 s during the experiment and the US was administered at 9.85 s in paired trials and co-terminated with the CS (Fig. 1A). At 7 s after CS onset, subjects performed an expectancy rating, which consisted of judging the likelihood of US delivery on a discrete scale. For this purpose, three symbols (−, ? and +) appeared underneath the fractal images for 2 s and participants rated the likelihood via button press according to the symbols’ meaning (“no shock”, “maybe shock”, “shock”). The intertrial interval varied randomly selleck kinase inhibitor between 9 and 11 s across trials. During the intertrial interval

a fixation cross was shown on the screen. The instructions were to concentrate on the images and complete the expectancy rating spontaneously in every trial. Subjects were aware that their responses did not influence the likelihood of US delivery. Before the experiment, subjects were familiarized with the task using different fractal images without US presentation. INCB024360 clinical trial Subjects were not informed about the two experimental phases prior to the experiment and the reversal stage started immediately after acquisition without announcement. Task presentation and recording of behavioural responses were performed with the software Presentation (Neurobehavioral Systems, Albany, CA, USA). Skin conductance responses (SCRs)

were recorded using a constant voltage system with Ag/AgCl electrodes placed on the hypothenar eminence of the left hand. Responses were amplified and digitized at 1000 Hz (CED 2502 and micro 1401, Cambridge Electronic Design, Cambridge, UK). A 3 Tesla system (TRIO; Siemens, Erlangen, Germany) equipped with a 32-channel head coil was used for acquisition of the fMRI data. Thirty-six transversal slices (slice thickness, 1.5 mm; no gap) were obtained in each volume using a high-resolution T2*-sensitive gradient echo-planar imaging sequence (repetition

time, 2680 ms; echo time, 30 ms; flip angle, 80°; field of view, 219 × 219 mm; in-plane resolution, 1.5 × 1.5 mm; parallel imaging with acceleration factor 2). Functional image coverage included the medial temporal Montelukast Sodium lobe, parts of the prefrontal cortex and brainstem areas. High-resolution T1-weighted anatomical images (1 × 1 ×1 mm³) were also acquired using a magnetization prepared rapid gradient echo sequence. We acquired four sessions consisting of between 210 and 250 volumes each, to sustain optimal quality of the high-resolution fMRI data. The sessions succeeded with the shortest possible latency (40–50 s) and the experimental presentation was not interrupted during scanner breaks in order to assure continuous learning unbiased by attentional changes caused by experimental breaks.

7% agar and containing 100 μL of overnight bacterial culture was

7% agar and containing 100 μL of overnight bacterial culture was spotted after solidification with 5 μL suspensions of the 16 isolated bacteriophages. The plates were incubated at 25 °C, and the occurring lysis was investigated after 18–24 h. Propagation of phages for genome isolation was initiated according to the double agar layer method (Adams, 1959). After incubation at 25 °C for 18 h, the top layers were collected and placed into 10 mL SM buffer. ATR inhibitor After gentle agitation for 4 h at 25 °C, 2 mL chloroform was added, mixed, and incubated at 4 °C for 18 h. The resulting suspension was decanted over the chloroform and centrifuged at 5000 g for 40 min at 10 °C to eliminate the bacterial cells;

the supernatant was centrifuged again at 16 000 g for 60 min at 10 °C to collect the phage particles. The pellet was resuspended in 500 μL EDTA (pH 8.0) and 500 μL TES (10 mM Tris–HCl, 10 mM EDTA, 2% SDS, pH 8.0). After vigorous vortexing

for 30 s, the solution was incubated for 30 min at 65 °C. The proteins were precipitated with protein precipitation solution (Sigma-Aldrich), incubated on ice for 5 min, and centrifuged at 15 000 g for 4 min at 10 °C. The nucleic acid was precipitated with 0.1 volume 3 M Na-acetate and 1 volume ethanol. The pellet was washed with 70% ethanol; dried, and resuspended in 20 μL TE buffer (10 mM Tris–HCl, 1 mM EDTA, pH 8.0). To determine the type of the genome nucleic acid, RNase and DNase treatments were carried out according to the manufacturer’s (Sigma) instructions. Restriction fragment pattern differences of the investigated phage DNAs were examined BTK signaling pathway inhibitor with 21 different restriction endonucleases: AatI, AluI, ApaI, BamHI, BcuI, BglI, BglII, Bsh1236I, ClaI, DraI, EcoRI, HaeIII, Hin6I, HindIII, KpnI, MspI, NotI, PstI, RsaI, SacI, TaqI (Fermentas, Thermo Scientific). The growth Baf-A1 supplier characteristics of the Bf7 were investigated using the double layer method

(Adams, 1959) on its host, incubated for 18–48 h at different temperatures (5, 10, 20, 25, 30, and 35 °C), and the resulting plaque numbers and morphologies were compared. The single-step growth curve experiments were carried out according to the protocol of Keel et al. (2002), with minor modifications. The LB liquid medium was supplemented with glucose (0.3%), CaCl2 (0.075 mM), MgSO4 (2 mM), and FeCl2 (0.004 mM) according to the suggestions of Sambrook et al. (1989), for higher phage titer. Exponential-phase culture of P. tolaasii 2342T was treated with bacteriophage solution to have a multiplicity of infection (MOI) of 0.06. To visualize phage morphology with transmission electron microscopy, phage plaques were picked and placed in SM buffer. Aliquots were mounted on a carbon-coated formvar film supported by a 300 mesh copper grid. Samples were negatively stained with 1% uranyl acetate and examined by a Zeiss CEM 902 electron filtering electron microscope.

Active TB needs to be excluded before considering treatment of la

Active TB needs to be excluded before considering treatment of latent infection, which is usually with isoniazid monotherapy for 6 months or isoniazid/rifampicin find more for 3 months. Starting HAART reduces the risk of reactivation of latent TB infection and is effective at reducing the incidence of new TB. We recommend that all HIV-positive patients should be offered HAART in line with the British HIV Association (BHIVA) treatment guidelines [2]. We recommend daily TB treatment whenever possible. Treatment

may be given 5 days per week, but should be intensively supervised. This option may be useful in hospital or other highly supervised settings. Three-times-per-week directly observed therapy (DOT) should only be given to patients who are stable and clinically well and where local logistics

enable this to be undertaken successfully. We do not recommend twice-weekly see more DOT for treatment of HIV/TB coinfected patients, especially in those with CD4 counts <100 cells/μL, as it has been associated with unacceptably high rates of rifamycin resistance. In cases where multiple drug resistance is not suspected, treatment should be started with four drugs (typically rifampicin, isoniazid, pyrazinamide and ethambutol) until sensitivities are known. We recommend a 6-month treatment regimen for drug-sensitive TB outside of the central nervous system (CNS). This is usually four drugs for 2 months, followed by isoniazid and rifampicin for a further 4 months (at least 182 doses of isoniazid and rifampicin and 56 doses of pyrazinamide and ethambutol in total). In drug-sensitive TB affecting Orotidine 5′-phosphate decarboxylase the CNS we recommend 9 months of treatment. This usually consists of four drugs for 2 months, followed by 7 months of isoniazid and rifampicin [3]. Drug-resistant disease should be treated only by specialists with experience in such cases, in line with NICE guidelines [1]. Careful attention should be paid to drug interactions between TB drugs, HAART and other therapy. Rifampicin is a powerful inducer of cytochrome 450 (CYP450)

and has effects on several metabolic pathways and P-glycoprotein (PgP). Rifampicin interacts with protease inhibitors (PIs), NNRTIs, chemokine (C-C motif) receptor 5 (CCR5) antagonists, and antimicrobials such as fluconazole. Rifabutin is a less potent inducer of CYP450 and may be used as an alternative to overcome some of these difficulties (for up-to-date drug interaction data go to Toxicity profiles of antiretrovirals and anti-tuberculosis drugs overlap and make it difficult to determine the causative drug. For example, rashes occur with NNRTIs, rifampicin and isoniazid. Isoniazid and stavudine both cause peripheral neuropathy. All patients on isoniazid should take pyridoxine to try and prevent this complication.

ART-CC is a carefully validated prognostic model based upon data

ART-CC is a carefully validated prognostic model based upon data from cohorts in Europe and North America [3,13,32]. It is focused on markers of HIV disease severity

and includes CD4 count (<50, 50–99, 100–199, 200–349 and ≥350 cells/μL), HIV-1 RNA of five log or more and the presence of AIDS-defining illness. For ‘non-HIV’ biomarkers we considered only: (1) clinical markers that are ordered as part of routine clinical management and (2) markers that have been previously demonstrated to be associated with mortality among patients with HIV infection. We employed previously validated specifications of these markers consistent with major organ system injury. For liver injury, we employed the Fibrosis Index (FIB) 4 [33]. FIB 4 uses aspartate and alanine transaminase (AST and ALT, respectively), Inhibitor Library cell line platelets and age to estimate likely liver fibrosis [FIB 4: (years of age × AST)/(platelets in 109/L × square root of ALT)]. Two thresholds of FIB 4 are recommended: >3.25, consistent with high risk for fibrosis/cirrhosis; and <1.45, consistent with low risk for fibrosis/cirrhosis. For renal injury, we employed the Modified Diet in Renal Disease (MDRD) estimation which uses age, race, gender and creatinine to estimate creatinine clearance [estimated Glomerular Filtration Rate (eGFR):

186.3 × (serum creatinine−1.154) × (age−0.203) × (0.742 for women) × (1.21 if African American)] [34]. Two levels of anaemia were defined: moderate and severe SP600125 chemical structure (haemoglobin 10-12 and <10 g/dL, respectively). Finally, we included a combined indicator variable for chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infection. We created a single indicator because 51% of those with chronic HBV infection also had HCV infection, and coefficients for HBV and HCV infections were similar in preliminary models. The Tolmetin ART-CC model also adjusts for two demographic factors: age ≥50 years and history of injecting drug use. Because our sample is older [3,13], we adjusted both models for age 50–64 and ≥65 years.

We did not have information available in Virtual Cohort on injecting drug use. As a proxy, we adjusted both models for a diagnosis of substance (drug or alcohol) abuse or dependence. We created a single indicator for substance abuse or dependence because 67% of those with a diagnosis of drug abuse or dependence also had a diagnosis of alcohol abuse or dependence [35] and coefficients in preliminary models were similar. Proportions were compared using the χ2 test. Medians were compared using the rank-sum test. Discriminations were compared using C statistics. The C statistic can be interpreted as the probability that any random pair of uncensored subjects in the data will be ranked correctly by the index with respect to their risk of mortality.

To this end we compared BOLD responses

To this end we compared BOLD responses Roxadustat cost evoked by search in the same parts of the VF, while having the eyes in different orientations relative to the head; conversely, by keeping non-eye-centred locations during search constant, while varying the position of search items within the VF. Our results suggest that both the IPS and the right FEF contribute to visual search by using eye-centred coding. Fourteen right-handed and one left-handed subject (mean age 27.1 years; six female, nine male; with normal or corrected-to-normal visual acuity, using scanner-compatible

glasses) participated in this study, which was approved by the Ethics Review Board of the Medical Faculty of Tübingen University. Hence, this study fully conforms with the code of ethics of the World Medical Association [Declaration of Helsinki; see Br Med J (July 1964), 818]. Each subject provided her/his written informed consent and was compensated financially for her/his participation. The subjects had to perform a task of covert

visual search in a mixed-block/event-related fMRI setting. Visual stimuli were presented onto a screen positioned frontoparallel to the subjects lying supine in the scanner. Using a beamer (NEC GT 950 1024 × 768 pixel) the stimuli were back-projected from outside the scanner room onto a mirror directing the beam parallel to the bore and centred onto a semi-opaque (diameter 60 cm) screen in the darkened scanner. The subjects were able to view the screen using a mirror system attached to the Alpelisib cell line Pregnenolone head-coil at a viewing distance of 70 cm. Stimulus presentations and data recording were controlled by a program written in LabView. Each trial consisted

of three epochs, as shown in Fig. 1B. After a randomly varied fixation period of 500–2500 ms [the white fixation point (diameter 0.35°) was either projected straight ahead, 10° to the right or left on the black screen], subjects were exposed to a search array in either their right or left visual hemifield. The array had a width of 3° (height 8°) visual angle and was placed with its centre at 5° eccentricity to the left or right of the fixation point. The array consisted of six ‘L’-shaped items (each 1.2° × 1.2°). A singular ‘L’ of conventional orientation, present in 50% of the trials, served as the target. The other items in the field, serving as distractors, were ‘L’s of non-conventional orientation obtained by mirroring a normal ‘L’ at one of the cardinal axes. Subjects had 3000 ms to decide whether the target was present in the array or not, while keeping fixation. At the end of this search period, the fixation dot disappeared and, at the same time, two response targets appeared on the vertical meridian, 4° above and below the centre.

The database is a depository of complete information on: the chem

The database is a depository of complete information on: the chemical structure of peptides; target species; target object of cell; peptide antimicrobial/haemolytic/cytotoxic activities; and experimental conditions selleck screening library at which activities were estimated. The dbaasp search page allows the user to search peptides according to their structural characteristics, complexity type (monomer,

dimer and two-peptide), source, synthesis type (ribosomal, nonribosomal and synthetic) and target species. The database prediction algorithm provides a tool for rational design of new antimicrobial peptides. dbaasp is accessible at “
“There is limited information on whether parasites

act as vectors to transmit bacteria in fish. In this trial, we used Ichthyophthirius multifiliis and fluorescent Edwardsiella ictaluri as a model to study the interaction between parasite, bacterium, and fish. The percentage (23–39%) of theronts fluorescing after exposure to E. ictaluri was significantly higher than control theronts (~ 6%) using flow cytometry. Theronts exposed to E. ictaluri at 4 × 107 CFU mL−1 showed a higher percentage (~ 60%) of fluorescent theronts compared to those (42%) exposed to 4 × 103 CFU mL−1 at 4 h. All tomonts (100%) carried the bacterium after exposure to E. ictaluri. Edwardsiella ictaluri survived and replicated during tomont division. Confocal microscopy demonstrated that E. ictaluri was associated with the tomont surface. Among theronts released from tomonts exposed to E. ictaluri, 31–66% were observed with attached E. ictaluri. Sixty percent of fish exposed to theronts treated with 5 × 107E. ictaluri mL−1 were positive for E. ictaluri at 4 h as determined by qPCR or fluorescent microscopy. Fluorescent E. ictaluri were observed on trophonts in skin and gill wet mounts of dead fish. This study demonstrated that Ich could vector E. ictaluri to channel catfish. In aquaculture systems, fish rarely encounter

a single pathogen. Most often, fish are concomitantly infected by multiple disease agents (Shoemaker et al., 2008). Parasitism has been demonstrated to enhance bacterial invasion where parasitic injuries serve as portals of entry Tacrolimus (FK506) (Buchmann & Lindenstrøm, 2002; Busch et al., 2003; Bandilla et al., 2006). Ahne (1985) reported that parasites Argulus foliaceus and Piscicola geometra served as mechanical vectors for spring viremia of carp virus (SVCV). Vijayan et al. (2005) reported that polychaete worms acted as vectors of white spot syndrome virus in the transmission of white spot disease to the shrimp Penaeus monodon. Cusack & Cone (1985) detected bacterial colonies on the surface of Gyrodactylus by scanning electron microscopy. However, they did not determine whether the bacteria were pathogenic to fish, and thus, the exact role of the bacteria was not clear.

001) Forty-eight per cent of children had etravirine mutation-we

001). Forty-eight per cent of children had etravirine mutation-weighted scores ≥4. There was a trend towards a higher rate of etravirine mutation scores ≥4 among children who received nevirapine than among those on efavirenz (52.8%vs. 31.0%; P=0.12). In the univariate analysis, there was no association between the duration of NNRTI treatment, the CD4 percentage, or plasma HIV selleck RNA and the risk of etravirine resistance. This study investigated the HIV resistance pattern in children with treatment failure on WHO-recommended first-line NNRTI-based ART. Eighty-five per cent of the children had resistance to lamivudine,

and about a quarter of the children had multi-NRTI resistance mutations conferring resistance to all NRTI drugs, which limit opportunities for recycling NRTIs as a component of the second-line PI-based regimen. Ninety-eight per cent of the children had at least one mutation related to NNRTIs, with half having high-grade etravirine resistance. A CD4 percentage <15% and an HIV RNA >5 log10 copies/mL at the time of genotype testing predicted multi-NRTI resistance. First-line NNRTI-based treatment failure is a major public health problem, especially in children, because of the limited availability of approved second-line

antiretroviral drugs and access to new drugs. Moreover, the lack of routine viral load monitoring in many resource-limited countries leads to delay in early detection of children who have virological failure. This causes accumulation of mutations within the NRTI and NNRTI drug classes until treatment failure is finally diagnosed

on the basis of clinical or immunological criteria [16]. Lapphra et al. reported that 8.4% of Thai children who started NNRTI regimens had treatment failure at 24 months [17]. Jittamala et al. [18] recently showed that 20% of Thai children had virological failure within 5 years of starting NNRTI-based regimens, with the majority failing in the first 12 months. These reports underscore the need for an understanding of resistance development, in order to design effective second-line regimens, especially if the availability of genotype testing is limited. Recently, the National Health Security Office, selleck products which provides ART to almost all HIV-infected Thai children, reported that 20% of HIV-infected Thai children are receiving second-line PI regimens. The regional Asian network, Treat Asia, which follows over 1000 children, also reported that 20% of children were on second-line ART [19]. The children in our study were from eight large paediatric HIV centres in Thailand. Similar to other studies on children from South Africa [6] and Thailand [8,18], extensive NRTI mutations were found. The rate of multi-NRTI resistance with at least four TAMs was as high as 23%, which limits the potential for recycling of NRTIs, including tenofovir.

Once the optimal IPTG concentration

Once the optimal IPTG concentration GSK-3 assay was obtained, additional cell-based assays were performed against a panel of known antibiotics of different chemical classes to obtain fold sensitization values [the ratio between the IC50 value (the concentration at which cell growth inhibited 50% compared with control) under noninduced condition and that of induced condition]. Based on the result of a time-course Sau3AI digestion (Fig. S1a, Supporting Information), the optimal partial digestion time was 4 h to generate DNA fragments of

appropriate size for library construction. Ligation mixtures were transformed into E. coli DH5α competent cells. Insert cloning efficiency analysis (Fig. S1b) indicated that the cloning efficiency for this library was 92%. To increase the randomness of the genomic DNA generated,

an alternative genomic library was constructed using a blunt-end producing restriction endonuclease (CviKI-1). The cloning efficiency of the CviKI-1-based library (termed Library C) was 90%. To screen for inducible growth inhibitory recombinant clones, transformants MAPK Inhibitor Library were grown overnight with chloramphenicol in the presence or absence of inducing IPTG. An example of screening plates and sensitive clones is shown (Fig. S1c). A total of 1500 confirmed IPTG-sensitive clones were mafosfamide obtained from screening 250 000 individual transformants. Only 675 of the 1500 confirmed clones were sequenced. An example of inducer-dependent inhibition of growth of asRNA clone PT113 is shown in Fig. S1d. Plasmid DNAs from a total of 675 confirmed inducer sensitive clones were sequenced. It was determined that enough clones were analyzed because more analysis leads to identification of duplicates, suggesting that the

phenotypic screening process under the condition scheme is approaching saturation. Among the sequenced clones, 134 separate clones contained insert DNA sequences derived from and in antisense orientation to known essential genes based on PEC database (Table 1). For most of the essential genes targeted by asRNAs, multiple gene silencing asRNA constructs were discovered, with rplF gene (encoding 50S ribosomal subunit protein L6) being ‘hit’ the most (17 times) (Table 1). Because many essential genes engaging in a cellular process are usually clustered in an operon, many essential operons are targeted by a multitude of asRNAs, especially the operons for ribosomal protein genes. For example, rplN operon that contains 11 essential genes was ‘hit’ by 17 unique asRNAs (Fig. 1a, with two asRNAs not shown owing to space limit). On an individual gene level, four unique asRNAs were found to target fusA gene (Fig. 1b), while another four to target rpoC gene (Fig. 1d).