Median nerve cross-sectional area (CSA) and flattening ratio (FR) at three different levels, proximal to tunnel inlet, at tunnel inlet and tunnel outlet, and flexor retinaculum thickness, were measured. Then, comparisons between ultrasonography and NCS were made. We assessed 180 wrists, of which 120 were electrophysiologically confirmed as CTS diseased hands and 60 nondiseased hands in 90 patients (83 women and seven men). The mean median nerve CSA at the tunnel inlet was 13.31 ± 3.23 mm2 in CTS diseased hands and 8.57 ± 0.82 mm2 in nondiseased hands. Post hoc comparisons between

the diseased and nondiseased hands demonstrated that the CSA at various levels of the median nerve were significantly greater in the CTS diseased hands than the nondiseased hands (P = 0.001). CSA at the tunnel inlet with a threshold of 9.15 mm2 gave the best diagnostic accuracy with a sensitivity and specificity of 99.2% Vorinostat datasheet and 88.3%, Sirolimus chemical structure respectively. The difference in cross-sectional area of the median nerve in mild, moderate and severe CTS was statistically significant. Ultrasonographic measurement of the CSA of the median nerve at the carpal tunnel inlet is useful in diagnosing and grading CTS. “
“Interleukin (IL)-22 regulates the pathogenesis of autoimmune diseases. The role of

IL-22+ T-cells in the pathogenesis of rheumatoid arthritis (RA) is unclear. This study aimed at examining the levels of plasma IL-22 and the frequency of IL-22+ CD4+ T-cells in patients with RA. A total of

30 RA patients and 18 gender- and age-matched healthy controls were recruited. Their peripheral blood mononuclear cells were isolated and stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin for 6 h. The frequency of IL-22+, interferon (IFN)-γ+ and IL-17A+ CD4+ T-cells was characterized by flow cytometry. The levels of plasma IFN-γ, IL-17A and IL-22, serum C-reactive protein (CRP), rheumatoid factor (RF), anticyclic citrullinated peptide antibody (CCP) and erythrocyte sedimentation rate (ESR) were measured. The frequency of IFNγ–IL-17A–IL-22+, IFNγ–IL-17A+IL-22+, and IFNγ+IL-17A–IL-22+ T-cells in CD4+ T-cells and the levels of plasma IFNγ, IL-17 and Non-specific serine/threonine protein kinase IL-22 in RA patients were significant higher than those in healthy controls. The percentages of IL-17A+IL-22+CD4+ T-cells were correlated positively with the frequency of Th22 or Th17 cells in the RA patients. The percentages of IL-22+CD4+ T-cells were correlated positively with the values of disease activity score (DAS28) in the RA patients. The percentages of Th22 cells were correlated positively with the levels of plasma IL-22 in the RA patients. Our data suggest that IL-22+CD4+ T-cells may contribute to the pathogenesis of RA and that therapeutic targeting of IL-22 may be valuable for the intervention of RA. “
“Adult-onset Still’s disease (AOSD) is a rare disease.

There are approximately 350 million hepatitis B carriers and about 33 million this website HIV-infected people world-wide [69,70]. As the routes of transmission for these infections are similar, there is a significant rate of coinfection in patients. Underlying HIV infection increases the chance of HBV chronicity [71]. There are no comprehensive data from the UK defining HIV/HBV coinfection rates. However, data from the EuroSIDA study [72] showed a 9.1% prevalence of HBsAg coinfection in participating northern European centres. In a survey of 100 UK clinics in 2004, the

dual HIV/HBV infection rate was estimated to be 3–10% of patients in 93% of clinics [73]. In many parts of Africa, HIV/HBV coinfection is common, as seen in South Africa (5%) or Malawi (20%) AG-014699 solubility dmso [74,75]. Recent

immigrants from Africa represent the largest group of newly diagnosed HIV-positive people in the UK [76] and therefore high coinfection rates are to be expected. High rates of HBV infection are also seen in IDUs and therefore HIV/HBV is relatively common in this group of patients [77] 4.1.2.1 The influence of HBV on HIV infection. The natural history of HIV infection does not seem to be influenced by hepatitis B [71,72,78] although there is an increased rate of antiretroviral-related hepatotoxicity, and immune-reconstitution hepatitis [79–81]. 4.1.2.2 The influence of HIV on HBV infection. Although the evidence remains conflicting, acute infection with HBV is more likely to be mild or asymptomatic in HIV-positive patients compared with those who are HIV-negative [82,83]. The rate of hepatitis B clearance is

also lower, with up to 20–40% of infected patients progressing to chronic (>6 Cediranib (AZD2171) months) infection [82,83]. Progression to liver cancer is more rapid, with HIV-positive patients with HBV infection developing liver cancer younger than patients with HBV infection alone [52, 82–84]. Once HBV infection is established, liver damage is immunopathic (the immune response to the virus causes most of the liver damage) so liver disease would be expected to be less severe in HIV-related immunosuppression. However, recent evidence suggests that alanine aminotransferase (ALT) and liver inflammatory scores in HIV coinfected patients are no different to those in HBV monoinfected patients [78]. At very high levels of viral replication, HBV may have a direct cytopathic effect. Coinfection with HIV is generally accompanied by an increase in HBV replication [78], which might explain the evidence for an increased rate of progression to cirrhosis and death [72,78,85,86] when compared with HBV monoinfected patients. There is also a reduction in the rate of natural clearance of HBeAg by about 60% in coinfected patients compared with HIV-negative patients [87]. However, there are reports of patients clearing chronic HBV infection with the recovery of CD4 cell count responses following ART [88,89].

This was emphasised within the findings of the focus group since there was much discussion on the role of the pharmacist on this process, with students also elaborating on what influenced their perspective, e.g. religion, ethics, etc.The undergraduate cohort will be the next generations of pharmacists and these results may evidence the need for the curricula

to tackle the issues of PAS and professionalism buy Bleomycin within practice. 1. Hanlon TRG, Weiss MC, Rees J. British community pharmacists’ views of physician-assisted suicide (PAS). J Med Ethics 2000;26: 363–369. Sonia Chand, Paul Rutter University of Wolverhampton, Wolverhampton, UK To ascertain what influences students to study pharmacy. Enjoyment of science was cited by many students as a main reason to study pharmacy. A lesser number of students saw pharmacy as a way to help people. Students associated selleck chemical pharmacy with good career opportunities and pay. There is little published work on why students decide to study pharmacy.1,2 Both Roller and Willis et al have measured the comparative influence

of extrinsic (e.g. income, status, good career opportunities) and intrinsic (liking science, desire to help people, and intellectual satisfaction) factors for studying pharmacy.1,2 These studies gave some insight into decisions made by students, however, the study by Roller focused on Australian students and the work by Willis et al, although UK-based, captured 3rd year student views; additionally both studies now lack currency, especially as pharmacy has changed in the last 10 years in response to the UK governments’ desire to better use the clinical skills of pharmacists. Therefore, this study aimed to update understanding on why first year students choose to study pharmacy. A survey comprising of open, closed and semantic differential scale questions was developed from conducting a literature search into similar studies. It was piloted

to all year groups at one School of Pharmacy (SOP) to determine its validity and reliability. Twelve SOP’s were invited to be involved in the study. pentoxifylline These were chosen as they represented varied curricula content ranging from predominantly science-based to practice-based programmes. Eight SOP agreed to participate. Each SOP disseminated study information and provided students with an email link to an electronic survey hosted by Survey Monkey®. Quantitative data was analysed using SPSS 16.0 and qualitative data was analysed using Nvivo 9 using content analysis. Ethics approval was obtained from the XX Ethics Committee at the University of XX and, where appropriate, from each SOP’s ethics committee. A total of 178 students fully completed the survey. Overall response rate was 15%.Individual SOP response rate ranged from 3.5–39%. The following represents the findings from the open ended question asking students to describe what influenced their decision to study pharmacy.

“Inadequate knowledge” included drinking bad water, eating bad food, dirt/dirty environment, malnutrition, weather or climate, too much sun/heat, insects, standing water, too much thinking/overworking/stress, dirty water, contaminated air, taking too much antimalarial medicines for prevention, and change of environment. The proportion of participants with “inadequate” or “unclear” knowledge was 152/292 (52%). selleck chemicals llc Travelers who received pre-travel advice were significantly more likely to demonstrate “inadequate or unclear knowledge” (OR 2.22, CI 1.13–4.38).

Perceptions about theoretical and personal risk of contracting malaria were compared in both the French and Dutch studies.10,11 The French researchers found that 87% of respondents knew it was possible to get malaria in the country they were visiting; however, only 49% considered themselves at personal risk. While there was no difference between those attending the pre-travel clinic and those visiting a travel agent in their general knowledge of the possibility of contracting malaria, there was a difference in perception of personal risk. Thirty-three per cent of those who had visited a travel agency believed themselves to be at high risk of malaria Veliparib order compared to only 7% of those who had visited a travel clinic (p < 0.05). In the study

of Dutch VFRs,11 perceived risk of catching malaria was assessed as either “high” or “not high.” Overall, 54% considered it to be high, 33% having sought pre-travel advice. To measure personal risk, participants were asked how dangerous the risk was for themselves, compared to specific risk groups (the definition of these groups was not provided). Forty-six per cent categorized the risk to themselves as “very dangerous. Two studies (in the Netherlands11 and the UK12) also provided

data on how participants believed they would be protected from malaria and these included perceptions such as sustained immunity,12 having had a malaria vaccine,11,12 and never having suffered from malaria previously.11,12 Biological factors, specifically Selleck Gemcitabine sickle-cell trait and/or G6PD deficiency, were also perceived as providing malaria protection.11 A reduced perception of personal risk was also found among participants in the London study who had been brought up in the UK. Among this group, some participants believed that malaria caught while visiting friends and relatives in an endemic country would result in only a mild illness.12 Both French and Dutch studies describe the proportion of travelers who intended to or had taken chemoprophylaxis.10,11 Surprisingly, 201/292 (69%) of Dutch VFRs and 171/191 (94%) of French individuals affirmed their use of chemoprophylaxis.

[1] Leptospirosis is now considered an emerging disease in travelers. On July 1, 2011, two Australian male tourists aged 25 and 26 years were admitted to the emergency department of the Careggi Hospital, Florence, Italy, reporting a 1-week history of fever with sudden onset of headache, myalgia, nausea, vomiting, and diarrhea. They had

no significant past medical or surgical history and they had recently traveled from Venice to Florence. At the time of admission they showed similar clinical signs and symptoms, mainly jaundice, conjunctival hyperemia, and muscle tenderness. Routine hematological and biochemical profiles were similar (Table 1). In both cases laboratory findings evidenced acute renal failure and hepatic impairment. Vital signs were normal. On auscultation, the heart sounds had no abnormalities RO4929097 supplier and air entry was equal on both lungs with occasional scattered wheezing. Results of neurological examination were normal. Electrocardiogram, abdominal ultrasound, and X-ray of the chest revealed no abnormalities. Asked about recent exposure to animals, mud, or potentially contaminated freshwater sources, the young tourists mentioned they had settled at a campsite near Venice 2 weeks earlier; because of the heat, they had both immersed their feet in the waters of a Venice canal close to Rialto Bridge. One of them had also swum in it for less than a minute without

any protection for the conjunctiva.

No skin lesions or trauma were observed at the time of possible infection, nor any swallowing of the canal water. The common Etoposide purchase history of exposure to possible contaminated water, along with hepatic and renal impairment, suggested the diagnosis of leptospirosis. However, blood and urine specimens were collected for culture and polymerase chain reaction Teicoplanin (PCR) was also evaluated. Serum samples were tested by the microscopic agglutination test (MAT). Intravenous ceftriaxone (2 g every 24 h) was empirically administered.[2] Adequate fluids and a diuretic infusion were also started. Both patients required daily hemodialysis for 5 days as a result of the severe renal injury. Blood and urine cultures had no growth. The PCR result was positive for leptospiral DNA in the urine of both patients. First collected serum sample results (approximately the tenth day of disease) were positive by MAT in both subjects with titers to serovars icterohaemorrhagiae of 1 : 1,600 and serovars copenhageni of 1 : 200 (serogroup icterohaemorrhagiae). Serovars icterohaemorrhagiae and copenhageni are commonly associated with rats as reservoir hosts.[3] Ten days later, the titer of 1 : 1,600 was confirmed in the first subject, while that of 1 : 200 of the other attained 1 : 3,200 (Table 1). The clinical picture progressively improved, restoring normal function of liver and kidney, and they were discharged after 2 weeks.

4 μL 25% glutaraldehyde followed by a 5 min centrifugation at 2000 g and washed 2–3 times in 1 mL PBS. Twenty-five microlitres of the samples were dropped on the slides and covered with poly-lysine-treated coverslips, and were

examined by differential interferential contrast (DIC, also named Nomarski) microscopy using a Nikon TE2000U fluorescence inverted microscope with a Nikon Plan Apo NA 1.4 100× objective. Images were captured using a Photometics CoolSnap HQ 12-bit CCD black and white camera and were analysed using Metamorph ver6.3 (Universal Imaging Corporation). The S. suis xerS gene and its GSK1120212 cell line difSL site were identified by sequence analysis (Le Bourgeois et al., 2007), amplified by PCR, and cloned into plasmid vectors. The binding activity of S. suis MBP-XerS to difSL was analysed by gel retardation assays. In binding reaction mixtures, increasing

quantities of MBP-XerS were added to 3.8 pM DNA with 1 μg (3.8 nM) polydIdC competitor. Three retarded bands were observed at protein concentrations of 3.43 nM (Fig. 1a) with stronger retarded bands observed with increasing concentrations of 17-AAG MBP-XerS, and with two of them very close to each other. No retarded bands were observed when using labelled non-specific DNA (data not shown). In addition, substrates with one of the two putative binding sites deleted were also constructed by site-directed mutagenesis and tested. Binding to the half-sites was much weaker, with the only clear band observed for the substrate with the left half of the binding site. At the same concentration of protein, binding was stronger to the full length site compared with the left half-site alone (Fig. 1). Baf-A1 purchase The ability of XerS to form covalent complexes with the difSL site was tested using suicide substrates with a nick introduced in the middle of the sequence either in the top (TN) or bottom strand (BN) (Fig. 2c). These substrates ‘trap’ recombination intermediates after recombinase-mediated cleavage close to the centrally

positioned nick, generating a small fragment which diffuses away, leaving the remaining phosphotyrosine-linked intermediate unable to complete the subsequent ligation reaction during strand transfer. The formation of covalent complexes was observed for both the top strand nicked and bottom strand nicked substrates, with a higher intensity for the bottom-nicked substrate (Fig. 2a). The covalent complexes were not observed using XerSY314F, an active-site tyrosine mutant that was constructed by site-directed mutagenesis (data not shown). The exact positions of XerS-mediated cleavage on difSL on either the top or bottom-nicked suicide substrates were determined using substrates with an FITC label placed at the 3′ end of the internal nick (Fig. 2c). A 5 bp fragment was observed after incubation of wild-type MBP-XerS protein with the top-nicked substrate and a 6 bp fragment was visible with the bottom-nicked substrate (Fig.

4 μL 25% glutaraldehyde followed by a 5 min centrifugation at 2000 g and washed 2–3 times in 1 mL PBS. Twenty-five microlitres of the samples were dropped on the slides and covered with poly-lysine-treated coverslips, and were

examined by differential interferential contrast (DIC, also named Nomarski) microscopy using a Nikon TE2000U fluorescence inverted microscope with a Nikon Plan Apo NA 1.4 100× objective. Images were captured using a Photometics CoolSnap HQ 12-bit CCD black and white camera and were analysed using Metamorph ver6.3 (Universal Imaging Corporation). The S. suis xerS gene and its PF-562271 in vitro difSL site were identified by sequence analysis (Le Bourgeois et al., 2007), amplified by PCR, and cloned into plasmid vectors. The binding activity of S. suis MBP-XerS to difSL was analysed by gel retardation assays. In binding reaction mixtures, increasing

quantities of MBP-XerS were added to 3.8 pM DNA with 1 μg (3.8 nM) polydIdC competitor. Three retarded bands were observed at protein concentrations of 3.43 nM (Fig. 1a) with stronger retarded bands observed with increasing concentrations of selleck kinase inhibitor MBP-XerS, and with two of them very close to each other. No retarded bands were observed when using labelled non-specific DNA (data not shown). In addition, substrates with one of the two putative binding sites deleted were also constructed by site-directed mutagenesis and tested. Binding to the half-sites was much weaker, with the only clear band observed for the substrate with the left half of the binding site. At the same concentration of protein, binding was stronger to the full length site compared with the left half-site alone (Fig. 1). Methocarbamol The ability of XerS to form covalent complexes with the difSL site was tested using suicide substrates with a nick introduced in the middle of the sequence either in the top (TN) or bottom strand (BN) (Fig. 2c). These substrates ‘trap’ recombination intermediates after recombinase-mediated cleavage close to the centrally

positioned nick, generating a small fragment which diffuses away, leaving the remaining phosphotyrosine-linked intermediate unable to complete the subsequent ligation reaction during strand transfer. The formation of covalent complexes was observed for both the top strand nicked and bottom strand nicked substrates, with a higher intensity for the bottom-nicked substrate (Fig. 2a). The covalent complexes were not observed using XerSY314F, an active-site tyrosine mutant that was constructed by site-directed mutagenesis (data not shown). The exact positions of XerS-mediated cleavage on difSL on either the top or bottom-nicked suicide substrates were determined using substrates with an FITC label placed at the 3′ end of the internal nick (Fig. 2c). A 5 bp fragment was observed after incubation of wild-type MBP-XerS protein with the top-nicked substrate and a 6 bp fragment was visible with the bottom-nicked substrate (Fig.

4 μL 25% glutaraldehyde followed by a 5 min centrifugation at 2000 g and washed 2–3 times in 1 mL PBS. Twenty-five microlitres of the samples were dropped on the slides and covered with poly-lysine-treated coverslips, and were

examined by differential interferential contrast (DIC, also named Nomarski) microscopy using a Nikon TE2000U fluorescence inverted microscope with a Nikon Plan Apo NA 1.4 100× objective. Images were captured using a Photometics CoolSnap HQ 12-bit CCD black and white camera and were analysed using Metamorph ver6.3 (Universal Imaging Corporation). The S. suis xerS gene and its selleck chemical difSL site were identified by sequence analysis (Le Bourgeois et al., 2007), amplified by PCR, and cloned into plasmid vectors. The binding activity of S. suis MBP-XerS to difSL was analysed by gel retardation assays. In binding reaction mixtures, increasing

quantities of MBP-XerS were added to 3.8 pM DNA with 1 μg (3.8 nM) polydIdC competitor. Three retarded bands were observed at protein concentrations of 3.43 nM (Fig. 1a) with stronger retarded bands observed with increasing concentrations of Venetoclax mw MBP-XerS, and with two of them very close to each other. No retarded bands were observed when using labelled non-specific DNA (data not shown). In addition, substrates with one of the two putative binding sites deleted were also constructed by site-directed mutagenesis and tested. Binding to the half-sites was much weaker, with the only clear band observed for the substrate with the left half of the binding site. At the same concentration of protein, binding was stronger to the full length site compared with the left half-site alone (Fig. 1). ID-8 The ability of XerS to form covalent complexes with the difSL site was tested using suicide substrates with a nick introduced in the middle of the sequence either in the top (TN) or bottom strand (BN) (Fig. 2c). These substrates ‘trap’ recombination intermediates after recombinase-mediated cleavage close to the centrally

positioned nick, generating a small fragment which diffuses away, leaving the remaining phosphotyrosine-linked intermediate unable to complete the subsequent ligation reaction during strand transfer. The formation of covalent complexes was observed for both the top strand nicked and bottom strand nicked substrates, with a higher intensity for the bottom-nicked substrate (Fig. 2a). The covalent complexes were not observed using XerSY314F, an active-site tyrosine mutant that was constructed by site-directed mutagenesis (data not shown). The exact positions of XerS-mediated cleavage on difSL on either the top or bottom-nicked suicide substrates were determined using substrates with an FITC label placed at the 3′ end of the internal nick (Fig. 2c). A 5 bp fragment was observed after incubation of wild-type MBP-XerS protein with the top-nicked substrate and a 6 bp fragment was visible with the bottom-nicked substrate (Fig.

5). The results for the perceptual matching study (i.e. A-A; see Fig. 5A) support the neural compensation hypothesis of cognitive reserve, as the activated regions underlying task performance

differed in the younger and older groups. The older group recruited the bilateral frontal superior gyri more than the younger one, as in the PASA phenomenon, even at the lowest attentional load level (i.e. three letters). In addition, the elderly participants were found to use neural compensation and neural reserve concurrently to cope with increasing attentional load (i.e. five letters) for perceptual GSK2118436 concentration processing. These results support a previous study (Townsend et al., 2006) which investigated auditory and visual selective attention and cross-modal attention shifts using fMRI. They showed age-related differences in BOLD responses. The most striking of these differences were bilateral frontal and parietal regions of significantly increased activation in older adults during both focused and shifting attention. These data suggest that this increased activation reflected not new recruitment but reliance on brain regions typically used by younger adults when task demands are greater. These patterns may reflect compensatory neural recruitment. The results for the

naming matching study (i.e. a-A; see Fig. 5B) indicate a load-dependent MDV3100 PASA, supporting the hypothesis that an enhanced compensatory mechanism is required for the most demanding find more condition (i.e. five letters). Thus, cerebral

reorganization of visual selective attention implies an intrahemispheric PASA phenomenon suggesting neural compensation. To cope with increased cognitive demand, neural reserve can also be recruited in basic perceptual processing (i.e. A-A; five letters), while the recruitment of compensation mechanisms increases in more complex processing (i.e. a-A; five letters). Taken together, these results suggest that the two neural mechanisms, compensation and reserve, are engaged in a flexible and adaptive manner, and are deployed depending on cognitive demand and on the nature of the required processing. Some of the evidence discussed here suggests that the functional reorganization of the brain that allows for the preservation of cognitive abilities takes many different forms, which cannot be universally predicted. Successful cognitive aging relies on neurofunctional flexibility, which enables the aging brain to cope with the challenges posed by declining neural structures. This flexibility is provided by dynamic neurofunctional adaptive mechanisms (a form of cerebral plasticity) that allow for the optimal engagement of the age-affected resources and recourse to the most advantageous distribution of cognitive processing and resources within the aging brain. This evidence suggests that neurofunctional reorganization in aging is based on a more flexible and adaptive neurofunctional process than had previously been proposed.

Interestingly, both FpTK1 and PdTK1, although Gram-negative, grouped together

with the usual Gram-positive TK1-like dNKs, rather than with the previously characterized Gram-negative ones (Fig. 1). Surprisingly for Gram-negative bacteria, in F. psychrophilum JIP02/86, we identified also one non-TK1 dNK (FpdNK), and in Polaribacter sp. MED 152, we found two non-TK1 dNKs, one of them representing a hybrid between non-TK1 and a sequence encoding HPPK (PdHPPK + dNK) (Table S2). The HPPK, 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase, catalyzes the attachment of pyrophosphate to 6-hydroxymethyl-7,8-dihydropterin to form 6-hydroxymethyl-7,8-dihydropteridine GSK J4 ic50 pyrophosphate. This is the first step in the three-step pathway that leads to 7,8-dihydrofolate. Similar hybrid genes were also found in several other bacteria belonging to Bacteroidetes class (Fig. S1). All the identified

dNKs genes were successfully amplified from genomic DNA (Table S1) and subcloned into the pGEX-2T expression vector. In addition, in order to test the significance of the HPPK domain for the phosphorylating activity of the Polaribacter sp. MED 152 PdHPPK + dNK hybrid, also a recombinant dNK without the HPPK domain Trichostatin A in vivo was constructed (PdHPPKΔdNK) (Tables S1 and S2). Initially, the substrate specificity of recombinant dNKs was tested in transformed TK1-negative Escherichia coli KY895 crude extracts (Table S2). dNKs phosphorylating activities were tested with all native dNs substrates: dT, deoxyadenosine (dA), deoxyguanosine (dG), and deoxycytidine

(dC). All assays were performed at 37 °C, except for FpTK1 and PdTK1. For these two enzymes, it was determined that 21 °C was the optimal temperature Pyruvate dehydrogenase lipoamide kinase isozyme 1 to measure their activity. In short, the subcloned TK1s and non-TK1s indeed represented active dNKs; however, the PdHPPK + dNK hybrid showed poor activity with dG, and the shortened recombinant enzyme PdHPPKΔdNK (without HPPK) also showed very low activity with dA, dC, and dG (Table S2). The hybrid proteins were not characterized further, and the function of the hybrid gene is so far unclear. All recombinant dNKs were expressed in E. coli BL21 and purified using affinity chromatography. The N-terminal GST fusion provided by the pGEX-2T vector was used as the affinity tag. Thrombin was used as a specific protease cleaving the GST tag from the kinase of interest, leaving only two extra amino acids (glycine and serine) at the N terminus. Afterward, pure recombinant protein was eluted from the GSH column. In the case of FpdNK and PddNK, we were not able to remove the GST tag from the dNK of interest; therefore, the whole GST fusion protein was eluted from the column and characterized. Purified dNKs were visualized by denaturing SDS-PAGE and Coomassie staining (Fig. 2). The size of the pure proteins was in reasonable agreement with theoretical molecular weights.