If there is a question about the patient’s capacity to make an informed decision, this should be assessed using Ku-0059436 purchase the principles in the Mental Capacity Act 2005 [28]. Patients presenting at the clinic may be at different stages of readiness to take therapy [29] and clinicians’ first task is to assess their readiness, by means of open questions rather than closed, before supporting and furthering patients’ decisions on therapy. However, if a patient presents in circumstances that necessitate starting ART immediately, for example with certain AIDS diagnoses or very low CD4 cell counts, then doctors should prescribe ART and provide support

for the patient’s adherence, especially through the first few weeks. Recognizing symptoms that patients attribute to ART side effects might avoid loss of adherence and deterioration of trust in the patient–provider relationship [30, 31].

A ‘perceptions and practicalities’ approach should be used to tailor support to meet the needs of the individual, to identify both the perceptual factors (such as beliefs about ART) and practical factors (such as capacity and resources) influencing adherence [8,32]. Supporting patients requires good communication not just between clinician and patient but also between all healthcare staff involved with their care, including those in their HIV services, their GP and any clinicians involved in management of co-morbid conditions. Patients should be offered copies of letters about them sent to their GP and other physicians. http://www.selleckchem.com/screening-libraries.html The advantages of HIV status disclosure to the patient’s GP should be discussed and considered best practice, as several situations require consensual clinical decision-making. A patient’s decision not to disclose their

status to their GP should, however, always be respected, subject to the clinician’s duty to protect vulnerable individuals. “
“Some fungi cause disease in humans and plants, while others have demonstrable potential for the control of insect pests. also In addition, fungi are also a rich reservoir of therapeutic metabolites and industrially useful enzymes. Detailed analysis of fungal biochemistry is now enabled by multiple technologies including protein mass spectrometry, genome and transcriptome sequencing and advances in bioinformatics. Yet, the assignment of function to fungal proteins, encoded either by in silico annotated, or unannotated genes, remains problematic. The purpose of this review is to describe the strategies used by many researchers to reveal protein function in fungi, and more importantly, to consolidate the nomenclature of ‘unknown function protein’ as opposed to ‘hypothetical protein’– once any protein has been identified by protein mass spectrometry.

[15] A total of 502 patients were included MK-2206 nmr in the four trials comparing rifaximin with placebo for prevention of TD (Figure 2).[15-18] One-hundred forty-two patients developed TD of which 41 were in the rifaximin group and 101 were in placebo group. The included trials were homogeneous (test for heterogeneity: p = 0.16, I2 = 42%), and the incidence of TD was significantly different between the rifaximin group and the placebo group (RR: 0.41, 95% CI: 0.30–0.56, p < 0.00001). NNT was four, which implied that four patients must receive rifaximin to avoid one case of TD. Seventy two of 404 patients in three

trials required antibiotic treatment for TD, 16 in the rifaximin group and 56 in the placebo group.[15, 18, 19] The included trials were not homogeneous (heterogeneity test: p = 0.11, I2 = 55%) so a fixed model was applied. The incidence of antibiotic treatment was significantly different between the rifaximin group and the placebo group (RR: 0.30, 95% CI: 0.18–0.49, p < 0.00001). NNT was five, which implied that one patient in every five would avoid

antibiotic treatment for TD. There were 197 patients involved in three trials comparing rifaximin with placebo in whom the incidence of MD could be evaluated.[15, 17, 18] The included trials were homogeneous (heterogeneity test: p = 0.25, I2 = 28%). Rifaximin was not associated HKI-272 nmr with significantly reduced incidence of MD (RR: 1.11, 95% CI: 0.78–1.59,

p = 0.55). There were 153 participants involved in two trials comparing rifaximin with placebo, reporting the incidence of TD in the third week after drug withdrawal.[16, 17] After eliminating the first 2 weeks of data regarding diarrhea, the data were not homogeneous (heterogeneity test: p = 0.97, I2 = 0%). There was no significant difference (p = 0.47) in the incidence of TD in the third week after drug withdrawal between the two groups. Enterotoxigenic E. coli was the major cause of diarrhea and MD during the 2 weeks of drug administration.[16, 18] There was no significant RG7420 difference between the rifaximin group and the placebo group in TD associated with diarrheagenic E. coli (ETEC or EAEC) (RR: 0.52, 95% CI: 0.24–1.09, p = 0.08). There was significant difference between the two groups in the incidence of unidentified pathogens associated with TD (RR: 0.37, 95% CI: 0.19–0.69, p = 0.002).[16, 17] All trials reported that there were no observed differences in adverse events between the rifaximin group and the placebo group. There was no clinically significant or serious adverse event in any of these studies.[15-18] There were no clinically relevant laboratory abnormalities reported.[16, 18] This meta-analysis shows an advantage of rifaximin over placebo in preventing TD. [Correction added on 3 October 2012, after first online publication: the phrase “protecting TD” was replaced with “preventing TD”.

More importantly, these results demonstrate that even when microorganisms are not using HER2 inhibitor the aerobic respiratory chain for growth, they are still killed by CO-RMs. This indicates that proteins other than the respiratory cytochrome c oxidase are targeted by CO. Davidge et al. (2009) reported that the growth of E. coli was impaired by CORM-3 but not by CO. In this study, exposure of aerobically grown E. coli cells to CORM-3 caused c. 50% inhibition of bacterial respiration due to the binding of CO to the terminal oxidases. Importantly, CORM-3 impaired growth of antibiotic-resistant strains of P. aeruginosa (Desmard et al., 2009). Table 2 summarizes the

microorganisms, and their conditions of growth, that have been shown to be killed by CO-RMs. The effects of CO on the genome-wide transcriptome this website profile has been analysed for cultures of E. coli grown aerobically and anaerobically in minimal medium salts with CORM-2, in glycerol (aerobically), and in glycerol/fumarate (anaerobically) with CORM-3 (Davidge et al., 2009; Nobre et al., 2009). In all cases, CO-RMs caused significant alteration of the mRNA abundance of a large number of genes (Fig. 2). Under aerobic conditions, CO-RM represses the transcription of E. coli genes involved in the citric acid cycle, respiration and iron homeostasis, whilst it up-regulates the expression of genes involved in general defence mechanisms, and in methionine, sulphur and cysteine metabolism. For E. coli grown

anaerobically in the presence of CO-RM, the genes involved in iron homeostasis are down-regulated, whereas those involved in zinc homeostasis and biofilm formation are induced. Furthermore, genes participating in protein homeostasis, oxidative stress, zinc and methionine metabolism, and general

defence mechanisms are up-regulated independently of Metalloexopeptidase the oxygen conditions in which E. coli is grown (Fig. 2). The transcription data acquired for E. coli grown aerobically with CO-RMs suggests that the respiratory chain may be hindered (Fig. 2). In accordance, P. aeruginosa treated with CORM-3 reduced oxygen less rapidly (Desmard et al., 2009). As blockage of the electron transport chain enhances the generation of ROS, the gene expression profile of E. coli in the presence of CO-RMs is expected to share similarities with its transcriptional response to hydrogen peroxide (Zheng et al., 2001; Zuckerbraun et al., 2007; Wang et al., 2009). In fact, the expression of a number of genes is affected similarly in cells treated with either of the two chemicals. They include the E. coli spy, encoding a periplasmic protein that is induced by envelope stress, the ibpA and ibpB genes, encoding two heat-shock proteins that are related to protein stability, hptX, coding for a heat shock protein, dnaK, dnaJ and hslO genes, encoding chaperones, and genes encoding proteins involved in sulphur metabolism such as sbp and cysWA (Zheng et al., 2001; Davidge et al., 2009; Nobre et al.

Most of these requests were ordered from the hospital’s emergency department for suspected insufficient serum concentrations. Antiepileptic drug monotherapy is still the most frequently employed therapeutic strategy in adult patients with epilepsy in keeping

with the standard therapeutic guidelines. Sodium valproate is commonly used for different types of seizures reflecting its wide spectrum of anticonvulsant potential. Newer AED utilizations are becoming increasingly popular in our subjects particularly as add-on with other standard AEDs. “
“Objectives  To determine statin usage pattern and evaluate whether new generation statins are actually needed by the patients receiving them. Methods  selleck chemicals llc This retrospective cohort included patients receiving first-time statins at a tertiary care hospital in Thailand. Using electronic medical records from 2005, its indication was determined based on history of coronary heart disease (CHD) and CHD-risk equivalents. The lipid profiles tested within 30 days prior to the first date of statins prescription were analysed. Each patient was assessed as

selleck to whether statin was needed based on low-density lipoprotein cholesterol (LDL-C) reduction capacity and lipid goals. Results  A total of 2479 first-time statin users was included. Ninety percent of the users received simvastatin, while 8% and 2% received atorvastatin and pravastatin respectively. More than half (58.0%) used statins for primary prevention, although all usage of atorvastatin was considered not needed. Considering the use of statin for secondary prevention to achieve the LDL-C goal of <130 mg/dl (3.37 mmol/l), more than 80% of atorvastatin users could be switched to simvastatin. Only 8% of simvastatin Galactosylceramidase usage would not be able to achieve this target. When

the LDL-C goal was <70 mg/dl (1.81 mmol/l), 40.2% simvastatin users was considered appropriate, while 58.6% needed atorvastatin to be prescribed. Conclusion  A substantial proportion of patients did not need statins therapy, particularly for primary prevention. In addition, atorvastatin use is mostly not needed except in patients requiring statins for secondary prevention to achieve the LDL-C goal of <70 mg/dl (1.81 mmol/l). The findings should prompt hospital policy makers to develop measures to ensure the proper use of statins in their clinical settings. "
“Objective  Most epilepsies are managed with anti-epileptic drugs (AEDs), but medication non-adherence has been frequently reported. Satisfying patient information needs has demonstrated improved adherence. Multi-professional working has been encouraged to provide cost-effective health services by using the most appropriate healthcare professional. Research has demonstrated that pharmacist-led consultations are acceptable to patients with other medical conditions and therefore may be appropriate for patients with epilepsy.

, 2007). It is thus likely that the increase in prefrontal activation for Acheulean–Oldowan reflects the greater temporal and relational complexity of Acheulean toolmaking actions, which, to a greater extent than Oldowan flaking, are organized into flexible and internally variable action chunks, such as ‘platform preparation’ vs. ‘primary flake removal’ (Pelegrin, 2005; Stout, 2011). No significant prefrontal activation BTK inhibitor libraries was observed for Oldowan–Control, in keeping with previous conclusions regarding the relative simplicity

of Oldowan action sequences (Stout & Chaminade, 2007; Stout et al., 2008). On this interpretation, the anterior inferior parietal cortex and the inferior frontal sulcus form a parieto-frontal circuit involved in representing episode-specific intentions, causal relations and multi-component action sequences during toolmaking observation. The apparent abstraction (Hamilton & Grafton, 2006; Badre & D’Esposito, 2009) of causal/intentional processing in this circuit may be compared with a proposed ‘intermediate’

level representing ‘intentions in action’ as goal-oriented sequences of motor commands and predicted outcomes (de Vignemont & Haggard, 2008). Varying expertise across subject groups was associated with qualitative shifts Navitoclax nmr in the set of brain regions activated in response to Acheulean compared with Oldowan stimuli (Fig. 4; Table 3). These differences suggest a functional reorganization (Kelly & Garavan, 2005) involving the adoption of different cognitive strategies for action understanding. Naïve subjects show activation in core motor resonance structures together with the ventral prefrontal cortex, as expected for a low-level strategy of novel action understanding

through kinematic simulation. Trained subjects show strong, statistically indistinguishable responses to both Oldowan and Acheulean stimuli, perhaps reflecting the particular social context and motivational set associated with training. Finally, Expert subjects display activation in the medial prefrontal cortex, a classic ‘mentalizing’ region, suggesting a relatively high-level, inferential strategy of intention reading. One Suplatast tosilate cluster exclusive to technologically Naïve subjects occurred in the pars opercularis of the left posterior inferior frontal gyrus (Fig. 4, left). Pars opercularis is another core component of the putative human mirror neuronal system (Rizzolatti & Craighero, 2004), which, in contrast with the performance-monitoring functions of the anterior inferior parietal cortex described above, is thought to be responsible for the generation of the kinematic models used to execute (Fagg & Arbib, 1998) or simulate (Carr et al., 2003; Grafton & Hamilton, 2007; Kilner et al., 2007) motor acts.

Interestingly, IAA addition upregulates genes encoding a type VI secretion

system (T6SS), a kind of secretion system that has been specifically implicated in bacterium–eukaryotic host interactions. Moreover, many transcription factors showed altered expression in the different treatments, indicating that the regulatory machinery of the bacterium is altered in response to IAA (Van Puyvelde et al., 2011). Increasing evidence indicates that NO is a key signaling molecule that is involved in a wide range of functions in plants (Creus et al., 2005; Molina-Favero et al., 2008). It has been demonstrated that NO plays an important role in auxin-regulated signaling cascades, influencing root growth and development (Pagnussat et al., 2003). NO is produced by A. brasilense Sp245 under aerobic SB431542 manufacturer conditions, mainly owing to the activity of periplasmic nitrate reductase (Nap) (Steendhoudt et al., 2001). A nap A. brasilense mutant produces only 5% of the NO produced by the wild type and is not able to promote lateral learn more and adventitious root formation and plant development like the wild type (Molina-Favero et al., 2008). The relationship

between NO and IAA production in A. brasilense is still to be elucidated. However, a recent study revealed that a nap mutant of A. brasilense possesses a reduced ability to induce root hair formation and nodulation by rhizobia in vetch roots. Moreover, vetch roots inoculated with this mutant secreted less nod gene inducers than roots inoculated with wild-type A. brasilense, and the indole content of the growth

solution of napA-inoculated plants was reduced at a lower rate than those of wild-type-inoculated plants (Star et al., 2011). A wide variety of taxonomically different groups of microorganisms within the Bacteria and Archaea domains produce intracellular homopolymers or copolymers containing different alkyl groups at the β position, described Oxymatrine as polybetahydroxyalkanoates (PHAs). These polymers are used as energy and carbon storage compounds (Madison & Huisman, 1999). In A. brasilense, PHAs are major determinants for overcoming periods of carbon and energy starvation (Fig. 2). Increased survival upon starvation in phosphate buffer was observed in A. brasilense Sp7 relative to a phaC (PHA synthase) mutant defective in PHA production (Kadouri et al., 2002, 2003, 2005; Castro-Sowinski et al., 2010) (Fig. 2). The abilities of A. brasilense phaC and phaZ (PHA depolymerase) mutants to tolerate and survive to various stresses, including UV-irradiation, heat, osmotic shock, desiccation, and oxidative stress, were significantly impaired as compared with wild-type cells (Kadouri et al., 2003, 2005). In addition, PHA accumulation in A. brasilense was shown to support chemotaxis, motility, and cell multiplication. Therefore, it is well established that production of PHAs in A.

05). When acetylene was added in conjunction with ethanol in the presence of mixtures of chlorinated alkenes or alkanes, no significant degradation was observed (Table 1). In the presence of either mixture, the microbial growth rate was significantly reduced as compared with that in the presence of ethanol

and acetylene, i.e., 0.14±0.03 and 0.09±0.04 day−1 for growth on ethanol and acetylene in the presence of chlorinated alkenes and alkanes, respectively, as compared with a growth rate of 0.28±0.0001 day−1 in the presence of ethanol and acetylene only Natural Product Library cost (Table 2). The overall growth of Methylocystis strain SB2 in the presence of these mixtures, however, as measured by OD600 nm, was not significantly different from growth in the presence

of ethanol and acetylene (Table 2). Here, it is shown that Methylocystis strain SB2 can degrade a variety of chlorinated hydrocarbons when grown on either methane or ethanol, and that this degradation is due to pMMO activity under both growth conditions. Specifically, the addition of acetylene, a specific inhibitor of pMMO, to Methylocystis strain SB2 grown on ethanol led to no degradation of any compound, but growth still occurred. Further, all the chlorinated hydrocarbons were, individually, potent inhibitors of the growth of Methylocystis strain SB2 on methane. With the exception of 1,1,1-TCA, however, individual chlorinated hydrocarbons had little effect on the growth of this strain on ethanol, indicating that competitive inhibition of pMMO by chlorinated hydrocarbons was at least partly Cediranib (AZD2171) see more responsible for the reduced growth of Methylocystis strain SB2 on methane. The data also indicated that

not only did the compounds act as competitive inhibitors of pMMO activity, some substrate toxicity was also evident, particularly when combinations of chlorinated hydrocarbons were added. Specifically, although very little degradation of 1,1,1-TCA, DCM, and CF was observed when these compounds were added together for both methane and ethanol-grown cells, growth was substantially reduced. Further, the addition of acetylene to ethanol-grown cells eliminates the possibility of product toxicity as pMMO was inactivated, but reduced growth rates of Methylocystis strain SB2 were still apparent on combinations of both chlorinated alkanes and alkenes, suggesting that the total concentration of chlorinated hydrocarbons is an important issue that can limit the overall methanotrophic activity at high levels. These data extend the previous finding that the facultative methanotroph Methylocystis strain SB2 can degrade chlorinated hydrocarbons when grown on acetate (Yoon et al., 2011) by showing that this strain can also degrade such compounds when grown on ethanol.

[1,6,19,20] The majority of rural healthcare providers are sole practitioners with a lack of professional support from their own profession and

other healthcare providers.[4,35] Given the complexity of the medication pathway, medication-related problems and errors may occur at any stage. The selleck compound Australian Commission on Safety and Quality in Health Care indeed identified that 2–3% of Australian hospital admissions are related to problems with medications (approximately 140 000 annual admissions), originating either in the community or in hospital, and costing about AUD$380 million per year in the public hospital system alone.[1] Researchers have argued that pharmacists have extensive knowledge of, and expertise in, medications, and should play a major role to promote QUM, ensure safe medication practices and support rural healthcare providers throughout the medication Protein Tyrosine Kinase inhibitor pathway.[26,35,44] A key problem, however, is a recognised shortage of pharmacists and pharmacy services in rural areas, limiting the potential for pharmacists to enhance medication services.[7,44,57] It has been reported that over half (75 of 116)

of Queensland’s public hospitals have no pharmacist on site, and less than one-quarter of these non-pharmacist sites (18 of 75) have limited outreach pharmacist support.[57] Many rural outpatient clinics and healthcare centres are serviced by sole nurses or health workers, who also undertake medication supply and stock control in these facilities. These facilities often do not have the capacity to employ pharmacists, or are not within the vicinity of a pharmacy service, and hence receive minimal input from pharmacists.[7,34,57] About one-third of Queensland’s public hospitals that do employ pharmacists

(15 of 41) are reportedly serviced by sole pharmacists.[57] It has been postulated that cost-shifting for public hospitals from state-based to Commonwealth-based mafosfamide management, as proposed as part of major PBS Public Hospital Pharmaceutical Reforms in Australia, would improve funding and therefore clinical pharmacy services in rural or regional hospitals.[43] Workforce studies have confirmed aging of the pharmacy workforce and high rates of sole pharmacy practice in rural areas, in both hospital and community settings.[7,28] Some of the contributing factors for the low rates of younger pharmacists in rural areas include the perceived higher workload and shortfalls in support (e.g. mentoring and training) systems in rural areas.[4,28,58] The limited pharmacy workforce restricts the provision of extended medication services or enhanced pharmacy services, meaning that rural pharmacists are often focused on core services such as dispensing and drug distribution, as well as pharmacy supervision and management.

TFB cells produced a few CT and MT within 24 h, but there was no significant relationship between the percentage of traps and the concentration of bacterial cells (Fig. 2). The number of traps increased significantly (P<0.05) within 24 h when the conidia of A. oligospora were cultured in different concentrations of Chryseobacterium sp. TFB cells with 20% bacterial cell-free culture filtrates (Fig. 2). The percentage of traps increased as the concentration of Chryseobacterium sp. TFB cells

increased from 0.33 to 3.0 × 107 CFU mL−1 Cell Cycle inhibitor and then decreased at the highest concentration of bacterial cells of 3.67 × 107 CFU mL−1. However, the highest concentrations of bacterial cells also caused conidia lysis (data not shown). When cultured with bacterial cells (1.67 × 107 CFU mL−1) in PDB dilutions (1 : 50) containing 5% bacterial cell-free filtrate, conidia of A. oligospora produced more MT and a few CT within 24 h (Fig. 3e–f and 4). With increased concentration of bacterial cell-free filtrates from 5% to 10%, the number of total traps, MT and CT all increased, with the number of MT increasing more than that of CT (Fig. selleck chemical 4). When the conidia were cultured in bacterial cells (1.67 × 109 CFU mL−1) with 20% cell-free supernatant, A. oligospora produced 50% CT at

24 h and 90% CT at 48 h. Most traps were on the long germination hyphae while near conidia (Fig. 3n–p), and some traps formed directly upon germination with minimal or no hyphal extension (Fig. 3l and m) and the CT have several loops (Fig. 3m). With increased concentration of bacterial cell-free supernatant from 30% to 40%, A. oligospora produced more typical CT (Fig. 3h–k) and few MT (Fig. 4). Conidia germination was inhibited when cultured in bacteria with more aliquots of bacterial cell-free supernatant (data not shown). In the negative control treatment, no traps formed even when conidia of A. oligospora were cultured for 1 month (Fig. 3d). With the addition of different nutrient levels to co-culture medium at the start of the experiment,

the percentage of conidia germination and trap formation increased within 24 h with the decreasing nutrient (Fig. S2). However, the percentage of conidia germination as conidial and MT decreased to when conidia were cultured in bacterial cells with dilution PDB (1 : 200) and sterile water. SEM observations revealed that Chryseobacterium sp. TFB cells attached to A. oligospora hyphae and traps (Fig. 5e–l) when A. oligospora conidia were cultured with bacterial cells (1.67 × 107 CFU mL−1) containing its cell-free culture filtrates (20%) in PDB dilution (1 : 50). There were no bacterial cells that attached to A. oligospora hyphae when A. oligospora conidia were cultured with bacterial cells in sterile water or PDB dilution (1 : 50) (Fig. 5b–d). SEM results suggested that bacterial cell-free filtrates facilitated its cells adhering on the surface of A. oligospora hyphae and bacteria attached to A.

, 2006; Park et al., 2007; Tamang et al., 2008; Carattoli, 2009; Strahilevitz et al., 2009). The aim of this study was to demonstrate the expression of an inducible acquired pACBL in S. marcescens and Escherichia coli isolates from the same patient. Moreover, as the E. coli isolate

showed reduced susceptibility to quinolones, plasmid-encoded quinolone resistance (PMQR) were also screened. Bacterial isolates were recovered from a urine specimen collected during nephrostomy in a 68-year-old patient who had initially undergone BCG instillation therapy and was later treated surgically by radical EPZ5676 mouse cyst-prostatectomy for a vesicle and ureteral transitional cell carcinoma. This patient carried an ileal conduit. Conjugation assays were performed using the broth mating method at 37 °C. Entinostat price Escherichia coli and S. marcescens isolates suspected to harbour pACBL were used as donor strains. As a recipient strain, we used the E. coli HB101 (UA6190), which expresses a green fluorescent protein marker and is resistant to rifampin, gentamicin

and kanamycin. Briefly, donor and recipient cells from exponentially growing cultures [3 h at 37 °C with agitation in Luria–Bertani (LB) media] were mixed with a donor/recipient ratio of 1 : 1 and incubated overnight at 37 °C. Transconjugants were selected on LB agar supplemented with ceftazidime (10 μg mL−1) and rifampin (100 μg mL−1) and were exposed to UV illumination. Isolates were identified using the API System 20E (bioMérieux, Marcy l’Étoile, France). The disc diffusion susceptibility test was performed on both donor and transconjugant strains, according to Clinical

Laboratory Standards Institute guidelines, using commercially available discs (Neo-Sensitabs, Rosco Diagnostica S/A, Taastrup, Denmark). The antimicrobial agents included were ampicillin, piperacillin, cephalotin, cefuroxime, cefotaxime, ceftazidime, cefepime, aztreonam, imipenem, cefoxitin, amoxicillin–clavulanic acid, piperacillin–tazobactam, nalidixic acid, ciprofloxin, sulphonamides, trimethoprim, trimethoprim–sulphamethoxazole, chloramphenicol, rifampin, tetracycline, gentamicin, kanamycin, tobramycin, amikacin and streptomycin. The inducible AmpC β-lactamase was suspected when antagonism between oxyimino-β-lactams and imipenem or cefoxitin was observed from on primary antibiogram plates. The presence of scattered colonies in the inhibition halo of cefoxitin, cefotaxime, ceftazidime and aztreonam was also examined (Mirelis et al., 2006). Antimicrobial resistance genes present in donor and transconjugant strains were studied. ampC genes were characterized using a previously described multiplex PCR (Pérez-Pérez & Hanson, 2002). Specific primers used to obtain the complete blaDHA-1 gene sequence were: DHA-1A 5′-CTG ATG AAA AAA TCG TTA TC-3′ and DHA-1B 5′-ATT CCA GTG CAC TCC AAA ATA-3′. PCR conditions were one cycle of denaturation at 95 °C for 5 min, followed by 30 cycles at 95 °C for 1 min, annealing at 55 °C for 1 min and elongation at 72 °C for 1 min.