Further, we calculated the median time between VL tests, the perc

Further, we calculated the median time between VL tests, the percentage of VL tests taken within 4 months from the previous VL test and the distribution of first VL>1000 copies/mL. We stratified the calculations by gender, race (Caucasian vs. non-Caucasian), age at HIV

diagnosis (<40 years vs. ≥40 years), route of HIV transmission (men who have sex with men vs. heterosexual vs. injecting drug users), partnership status (reporting living in a stable partnership vs. reporting not living in a stable partnership), sexual behaviour (practising check details safe sex vs. practising unsafe sex), calendar year of HAART initiation, number of periods with VL<51 copies/mL (first episode of VL<51 copies/mL vs. later episodes of VL<51 copies/mL) and number of consecutive months with VL<51 copies/mL. In a subanalysis, we performed all the above stratifications on patients diagnosed with HIV after 1 January 2000. We tested for robustness by repeating our calculations with cut-off values for risk of transmission of HIV of 500 and 1500 copies/mL. The study was approved by the Danish Data Protection Agency. spss statistical software,

version 15.0 (Norusis; SPSS Inc., Chicago, IL, USA) was used for data analysis. We identified 2680 patients with a total of 47 895 VL tests performed XL765 nmr in the period 2000–2007. The median time between tests was 0.25 years [interquartile range (IQR) 0.21–0.31]. Of the tests, 81.2% were taken within 4 months of the previous test. The median VL at first VL>1000 copies/mL was 28 600 copies/mL (IQR 3812–1 000 000 copies/mL). A total of 182 (6.8%) of the study subjects died during follow-up, 33 (1.2%) were lost to follow-up and 37 (1.4%) emigrated. Overall, 1998 (74.6%) of the patients were male and 2106 (78.6%) were Caucasian.

The median age at time of HIV diagnosis was 34.3 years (IQR 28.1–42.5 years). Regarding route of transmission, 1250 (46.6%) were men who have sex with men, 1078 (40.2%) reported having been infected heterosexually and 203 (7.6%) reported infection through injecting drug use. Of the 1010 (37.7%) patients with available data on civil status, 540 (53.5%) Sinomenine reported that they were living with a partner. Eight hundred and thirty-one patients (31.0%) were diagnosed with HIV infection on or after 1 January 2000. Data on sexual behaviour were available for 1002 (37.4%) patients and 780 (77.8%) patients reported that they practised safe sex. The observation time (as defined above) for the population was 9347.7 years, during which the patients were at risk of transmitting HIV infection for 56.4 years. The overall percentage of time at risk of transmitting HIV was therefore 0.6% (95% CI 0.5–0.8%). The percentage of time at risk of transmitting HIV stratified by gender, race, age at HIV diagnosis, route of HIV transmission, status of partnership and sexual behaviour is shown in Table 1 and differs very little between the groups.

Further, we calculated the median time between VL tests, the perc

Further, we calculated the median time between VL tests, the percentage of VL tests taken within 4 months from the previous VL test and the distribution of first VL>1000 copies/mL. We stratified the calculations by gender, race (Caucasian vs. non-Caucasian), age at HIV

diagnosis (<40 years vs. ≥40 years), route of HIV transmission (men who have sex with men vs. heterosexual vs. injecting drug users), partnership status (reporting living in a stable partnership vs. reporting not living in a stable partnership), sexual behaviour (practising find more safe sex vs. practising unsafe sex), calendar year of HAART initiation, number of periods with VL<51 copies/mL (first episode of VL<51 copies/mL vs. later episodes of VL<51 copies/mL) and number of consecutive months with VL<51 copies/mL. In a subanalysis, we performed all the above stratifications on patients diagnosed with HIV after 1 January 2000. We tested for robustness by repeating our calculations with cut-off values for risk of transmission of HIV of 500 and 1500 copies/mL. The study was approved by the Danish Data Protection Agency. spss statistical software,

version 15.0 (Norusis; SPSS Inc., Chicago, IL, USA) was used for data analysis. We identified 2680 patients with a total of 47 895 VL tests performed http://www.selleckchem.com/products/H-89-dihydrochloride.html in the period 2000–2007. The median time between tests was 0.25 years [interquartile range (IQR) 0.21–0.31]. Of the tests, 81.2% were taken within 4 months of the previous test. The median VL at first VL>1000 copies/mL was 28 600 copies/mL (IQR 3812–1 000 000 copies/mL). A total of 182 (6.8%) of the study subjects died during follow-up, 33 (1.2%) were lost to follow-up and 37 (1.4%) emigrated. Overall, 1998 (74.6%) of the patients were male and 2106 (78.6%) were Caucasian.

The median age at time of HIV diagnosis was 34.3 years (IQR 28.1–42.5 years). Regarding route of transmission, 1250 (46.6%) were men who have sex with men, 1078 (40.2%) reported having been infected heterosexually and 203 (7.6%) reported infection through injecting drug use. Of the 1010 (37.7%) patients with available data on civil status, 540 (53.5%) oxyclozanide reported that they were living with a partner. Eight hundred and thirty-one patients (31.0%) were diagnosed with HIV infection on or after 1 January 2000. Data on sexual behaviour were available for 1002 (37.4%) patients and 780 (77.8%) patients reported that they practised safe sex. The observation time (as defined above) for the population was 9347.7 years, during which the patients were at risk of transmitting HIV infection for 56.4 years. The overall percentage of time at risk of transmitting HIV was therefore 0.6% (95% CI 0.5–0.8%). The percentage of time at risk of transmitting HIV stratified by gender, race, age at HIV diagnosis, route of HIV transmission, status of partnership and sexual behaviour is shown in Table 1 and differs very little between the groups.

005 Although the threshold did not reach our criteria, these res

005. Although the threshold did not reach our criteria, these results are compatible with the ROI analysis. As the order of the sessions was fixed, the observed results in the comparison between the random and within-/between-group omissions might include the influence

of adaptation or fatigue in general. In order to evaluate such influences, we created buy KU-60019 the reconstruction maps for the brain activity elicited by the L tones and conducted two-sample t-tests between the random sequence and group sequence. If such mental health conditions affected the omission-related response, it should also be shown in the brain activity elicited by the L tone. However, this analysis did Aloxistatin clinical trial not show any significant result in both subject groups, indicating that the obtained results for the omission-related response were not due to adaptation or fatigue. The

ability to integrate sound features is necessary to perceive a sequence of tones as a perceptual group, which might be a basis for auditory perception (Winkler et al., 2009; Bendixen et al., 2012). Many studies have indicated that pre-attentive processing of perceptual grouping works well when the ISI between tone stimuli is less than 200 ms (Yabe et al., 1997; Sussman et al., 1999; Deike et al., 2004; Micheyl et al., 2007). In the present study, we investigated the attentive processing of perceptual grouping using auditory stimuli with an ISI longer than 200 ms and the effect of musical experience. The results indicate that the regular pattern of the tone sequence had impacts on both the behavioral performance and neural response elicited by the omission. In addition, we found that musicians showed the right IPL for the processing of sound omission, whereas non-musicians showed the left STG. The results and possible interpretations are discussed in the following sections. Several psychological studies have shown that the perceptual grouping of stimuli affects behavioral performance. For example, detection Immune system or recognition is faster and more accurate for target stimuli that are inconsistent with the grouping

structure of stimuli, compared with consistent target stimuli (Idson & Massaro, 1976; Jones et al., 1982; Mondor & Terrio, 1998). This evidence is consistent with the results of the present study, which showed that detection of omission in the random sequence was faster than that in the group sequence. In addition, the subjects in the present study reported recognising the group sequence as a repetition of the ‘LLS’ pattern. Therefore, we believe that the perceptual grouping successfully occurred in the group sequence in the present study. The predictive coding theory suggests that the auditory system continuously searches for regularities to organise a perceptual unit within a tone sequence (Winkler et al., 2009; Bendixen et al., 2012).

Among these compounds, 2,4-diacetylphloroglucinol (2,4-DAPG) prod

Among these compounds, 2,4-diacetylphloroglucinol (2,4-DAPG) produced by some Pseudomonas spp. is of particular significance for the suppression of root diseases (Keel et al., 1996; Haas & Defago, 2005). The antibiotic 2,4-DAPG is a polyketide compound with antifungal, antibacterial, antihelminthic and phytotoxic activities (Keel et al., 1992; Dowling ABT-263 purchase & O’gara, 1994). The genes involved in the biosynthesis of this antibiotic cloned from several Pseudomonas strains include four structural

genes, phlA, phlC, phlB and phlD, which are transcribed as a single operon (phlACBD) (Fenton et al., 1992; Bangera & Thomashow, 1996, 1999; Wei et al., 2004a). A specific transcriptional regulator gene, phlF, is localized upstream of the phlACBD operon and transcribed in the opposite direction (Abbas et al., 2002). Intensive

studies on the regulation of 2,4-DAPG production in recent years have revealed a number of transcriptional and post-transcriptional elements. Besides PhlF, other identified regulatory elements include the two-component system GacS/GacA (Haas & Keel, 2003), sigma factors RpoS (Sarniguet et al., 1995), RpoD and RpoN (Schnider et al., 1995; Péchy-Tarr et al., 2005), the H-NS family regulators MvaT and MvaV (Baehler et al., 2006), the translational repressor proteins RsmA and RsmE (Heeb et al., 2002; Reimmann et al., 2005), the oxidoreductase DsbA (Mavrodi et al., 2006) and the resistance-nodulation-division efflux pump EmhABC (Tian et al., 2010). Quorum learn more sensing (QS)

is a process of cell-to-cell communication that enables bacterial populations to collectively control gene expression and thus coordinate group behaviors (Miller & Bassler, 2001). In many Gram-negative bacteria, selleck chemical the QS system is based on the function of two proteins that belong to the LuxI-LuxR family of transcriptional regulators. The LuxI protein synthesizes N-acyl-homoserine lactone (AHL) signaling molecules that can diffuse through the cell envelope. AHLs bind to the transcriptional regulator LuxR, forming a complex that plays an important regulatory role in a diverse array of physiological activities (González & Keshavan, 2006; Keller & Surette, 2006). QS has also been implicated in the interaction between plants and plant growth-promoting rhizobacteria. For example, the PhzI–PhzR QS system regulates the biosynthesis of the phenazine antibiotic in the plant-beneficial bacterial strains Pseudomonas aureofaciens 30-84 (Pierson et al., 1994) and Pseudomonas chlororaphis PCL1391 (Chin-A-Woeng et al., 2001). A second QS system in strain 30-84, CsaI-CsaR, which does not influence phenazine production, is involved in rhizosphere competitiveness and biosynthesis of cell-surface components (Zhang & Pierson, 2001).

3%, p < 0001) compared with those born in North Africa Overall,

3%, p < 0.001) compared with those born in North Africa. Overall, 135 (21.3%) pilgrims had traveled and planned to travel outside France, both before and after the pilgrimage. Our results show a complex pattern of international travel in French pilgrims participating in the Hajj of 2010. Two-thirds of them underwent a trip to their country of origin in North Africa, 1 to 4 months before traveling from France to Saudi Arabia and a quarter planned to go back to North Africa after a short stop-over in France, following the Hajj. Deforolimus manufacturer This reflects

France’s past colonial history in Algeria, Morocco, and Tunisia and the post-colonial migrations. Therefore, French pilgrims arriving to Saudi Arabia may both present with long incubation communicable diseases, acquired in North Africa and short incubation infections acquired in France. In case of acquisition of communicable diseases during their stay in Saudi Arabia, French pilgrims will have the potential to spread infectious disease agents not only in France, but also in North Africa. This was particularly worrying during the Hajj of

2009 regarding the risk of spread of influenza A H1N1 09.6 Collaborative sentinel surveillance networks monitoring disease trends among travelers offer valuable tools for evaluating travel health issues. However, the major multinational sentinel networks addressing travel health issues globally (GeoSentinel and EuroTravNet) are mainly based in industrialized countries and do not include sites in North Africa.7,8 The EuroTravNet center in Marseille captures only few cases of Hajj-associated selleck infectious diseases in returned French

pilgrims, although cohort studies have demonstrated that most pilgrims Branched chain aminotransferase departing from Marseille get ill during their stay in Saudi Arabia.9 This is due in part to the mild nature of Hajj-associated diseases that are not likely to be seen at a specialized clinic, but also to the fact that a significant proportion of travelers may exhibit symptoms in North Africa rather than in Marseille. Therefore, surveillance of Hajj-associated infectious diseases in French pilgrims should be coordinated between France and North African countries. In this perspective, collaboration with EpiSouth network, a recently born network aiming to improve communicable disease surveillance in the Mediterranean area could be useful.10 Our study is limited to a small cohort of pilgrims from one large city in France and although it is a tradition in Muslim communities that many pilgrims travel after Hajj in the Middle East and Indian subcontinent,6 our results cannot be extrapolated to all pilgrims. Better linked surveillance for travelers, including pilgrims to the Hajj, is needed by health information system development such as real time electronic reporting, rapid data collection and post-event reporting using mobile phone technology and social networking, and rapid laboratory testing where possible to improve outbreak detection and control.

graminis or to P betae None showed close identity to P gramini

graminis or to P. betae. None showed close identity to P. graminis type II despite

this ribotype being present in both soils (Ward et al., 2005; Lyons et HSP inhibitor al., 2008). Although temperate ribotypes of P. graminis have been shown mainly to infect monocotyledonous plants, P. betae and tropical isolates of P. graminis have been shown to infect dicotyledonous plants (Barr, 1979; Ratna et al., 1991; Barr & Asher, 1992; Legrève et al., 2000). The observation of spores in the root hairs of the Arabidopsis ecotype Ler-0 plants is interesting as Polymyxa spp. are not routinely reported infecting root hairs, although this has been observed infrequently (M. Smith & M.J. Adams, unpublished data). Because this is a new and distinctive host, it is not unreasonable to expect that that the localization of Polymyxa within the plant or aspects of its morphology might differ. This could result for example from spatial constraints within the cells. There is support for this from anatomical studies of P. graminis infection in sorghum and wheat (Littlefield et al., 1997). Unfortunately, we cannot confirm absolutely that the structures observed in the roots of the Arabidopsis Crizotinib molecular weight plants correspond to the Polymyxa detected using molecular methods. In hindsight, we should have

selected infected root tissue before DNA extraction to provide additional support for this, but conclusive proof would require a technique such as laser capture microdissection (Day et al., 2005).

These techniques are technically challenging and have rarely been successfully used in these types of study. There are problems associated with the use of soil to infect the plants rather than G protein-coupled receptor kinase resting spores or zoospores from previously characterized Polymyxa isolates. There is a possibility of detection of Polymyxa from soil adhering to the root, which could confuse the issue of whether detection in the plant has occurred. However, the roots were washed thoroughly before use and this was facilitated by growth in a mixture of soil and sand (1 : 2), rather than soil alone. Also, from our previous experience of this system, we feel that it is unlikely that loosely attached Polymyxa spores would be responsible for the detection. Infection using Polymyxa-infected material would also have been superior in that it would have allowed a demonstration of Koch’s postulates. However, it is generally more difficult to infect plants using zoospores or resting spores, than using soil and we felt that, to establish the system, it would be better to bait plants with the mixture of ribotypes that are present in the soil, rather than test individually zoospores/resting spores from a wide range of different isolates, some of which may not be well adapted to the new host.

IL-13 inhibits Th17 cell development in dendritic cells via down-

IL-13 inhibits Th17 cell development in dendritic cells via down-regulation of Th17 stimulatory cytokines (IL-1, IL-6 and IL-23).[46] Despite the inhibitory effect of GATA-3 on Th17 development, it seems that GATA3 probably promotes Th17 development through inhibition of IL-2, STAT1 and suppressors of cytokine signaling 3 (SOCS3).[47] IL-2 is a T cell growth factor that is critical for Treg development. It effectively inhibits Th17 cell development. Two pivotal transcription factors that BAY 73-4506 cell line mediate IL-2 signaling are STAT5a/b. Therefore IL-2 or STAT5 deficiency is associated with

inhibitory effects of Tregs and expansion of Th17 cells.[48-51] The transcription factor Ets-1, which is a positive regulator of Th1 development, is another negative regulator for Th17 development. Ets-1 deficiency leads to increased Th17 differentiation and promotion of IL-22 and IL-23R messenger RNA (mRNA) levels in response to IL-6 and TGF-β1. It seems that the inhibitory effect of Ets-1 on Th17 cells is through enhancing IL-2 production.[52] In a recent report,

it has been shown that microRNA mir-326 can bind to and prevent translocation of Ets-1 mRNA. Thus, microRNAs can promote Th17 development through inhibition of the Th17 inhibitor, Ets-1.[11-58] It should be noted that the transcriptional repressor protein BCL-6 regulates T cell differentiation BGJ398 by repressing Th2 cells and enhancing follicular Th cells. It is proposed that BCL-6 enhances Th17 differentiation through suppression of Th2 differentiation.[54] Th17 cells are the dominant

pathogenic cellular component in autoimmune inflammatory diseases, including RA.[55] Although the importance of Th17 cells in animal models of arthritis is unquestionable, there are only limited data on the role of Th17 cells and related cytokines in human arthritic diseases. In addition, the characteristics of human Th17 cells have not been fully defined, and there seems to be substantial differences between human and mouse Th17 cells.[56] Functionally, Th17 cells contribute to host defense by having a role in protection against extracellular bacteria. However, their activities are also pivotal in the development of autoimmune diseases under pathologic conditions.[57] The identification of Th17 and IL-17 as a powerful pro-inflammatory cytokine, have Endonuclease focused attention on the role of Th17 cells in RA and other immune-mediated diseases, such as psoriasis, Crohn’s disease and multiple sclerosis.[5, 58] The hyperfunction of Th17 cells is associated with autoimmune diseases, due to the hypersecretion of the pro-inflammatory cytokine IL-17.[59] Studies in rodents, mammalian cell culture systems, as well as clinical settings, support a specific role for IL-17 in promoting RA.[60] Additional supporting evidence came from IL-17 knock-out animals that failed to develop collagen-induced arthritis (CIA).

To a great extent, the explanations of the participants about pos

To a great extent, the explanations of the participants about possible disease aetiology are focused on stress, immigration and psychological well-being. Although many participants perceived that their own efforts did not have much impact on their health status, our study revealed a large diversity in the responses of non-Western immigrants,

particularly regarding the importance of their own efforts on their health status. “
“Welcome to this first issue of 2014. The New Year is always a time to reflect and to think back on the past and to make resolutions for the future. As I write this, I look back at the year 2013 and see that once again we have had an increased rate of submission to IJPP, a real marker of the Luminespib mouse strength and success of our discipline. The downside for some, perhaps, is that the landscape is more competitive and on the Editorial side, we increasingly have to make Opaganib hard choices and we are not always able to accept strong and interesting papers especially if they do not quite match others for topicality or novelty. The IJPP provides a window on the state-of-the-art of practice, research and education in the field of pharmacy

and medicine use. The way in which these three move forward together is exemplified in the papers that we publish. For example, with respect to extended pharmacy practice, in this issue alone, we have articles on pharmacist prescribing, interactions with general

practitioners and improving the management of warfarin, and enhanced roles for pharmacy with patients such as providing motivational interviews for drug misusers, and considering what happens when increasingly potent medicines are made available over the counter. The articles in this issue also show how practice research is advancing with encouraging signs of increased sophistication in research methods including multidisciplinary working with other disciplines, such as psychology, and better use of rigorous Non-specific serine/threonine protein kinase research designs including mixed methods approaches and understanding how we evaluate pharmacy roles informed by the Medical Research Council framework for complex interventions. We are also strong on the education front. Papers illustrate how integral good training is to research studies which evaluate the outcomes of pharmacists undertaking new roles. Educational researchers are also looking critically at assessments of students to ensure that they are both challenging and discriminatory. So if we are doing so well what are our New Year resolutions for 2014? It goes without saying that that there is nothing that cannot be improved. It would be nice to think that our relatively small profession is currently punching above its weight, and we need to continue to do that in a time of continued financial constraints limiting resources for both research and service delivery.

The results demonstrate the potential of GFP labeling for protein

The results demonstrate the potential of GFP labeling for protein functional characterizations in Xac, and, in addition, the Xac mutant strain labeled at the septum constitutes a biological model for the exploration of antibacterial compounds able to inhibit cell division in this plant pathogen. Xanthomonas

citri ssp. citri (Schaad et al., 2005, 2006) (also known as Xanthomonas axonopodis pv. citri or Xac) is a Gram-negative, plant-pathogenic bacterium that affects most citrus species and is the causal agent of citrus canker, a very economically important disease of citrus plants worldwide. An effective control for this disease is inexistent, and a more detailed understanding of the biology of the etiological agent may contribute substantially toward the development of strategies to prevent and control infection. A major effort to accomplish http://www.selleckchem.com/products/jq1.html this task was the elucidation of the genome sequence of Xac (da Silva et al., 2002), which has stimulated a number of molecular studies using Xac as model microorganism,

and yet, little information is available regarding technical methods that could enhance its proteome exploration (Galvao-Botton et al., 2003; Mehta & Rosato, 2003; Alegria et al., 2004, 2005; Cernadas et al., 2008). Our main interest focused on the characterization of some essential biological processes of Xac, more specifically those Selleck Maraviroc involved with chromosome segregation and cell division. A common feature of such bacterial systems is that they are usually composed of proteins sharing little homology to their functional analogues in more derived eukaryotes; therefore, these proteins constitute ideal targets for antimicrobial drug development and pathogen control (e.g. Gitai et al., 2005; Pan et al., 2006; Haydon et al., buy Hydroxychloroquine 2008; Beuria et al., 2009; Kapoor & Panda, 2009). However, to undertake protein functional studies with/in Xac, we were limited

by the lack of biological tools developed and/or accessible for this purpose. Here, we describe a protein expression system dedicated to Xac, which can also be used for the subcellular localization of the green fluorescent protein (GFP)-labeled factors in this pathogen. We used the system to characterize a hypothetical protein of Xac that shares significant homology to the FtsZ-stabilizing factor ZapA, originally described in Bacillus subtilis (ZapABsu) (Gueiros-Filho & Losick, 2002). Furthermore, we show that the disruption of the α-amylase gene, the site of plasmid integration into the Xac chromosome, does not alter its pathogenesis. The Xanthomonas citri ssp. citri used was the sequenced strain (da Silva et al., 2002), formerly designated X. axonopodis pv. citri strain 306 (IBSBF 1594). Escherichia coli strain DH10B (Invitrogen) was used for cloning. Escherichia coli was cultivated at 37 °C in a Luria–Bertani (LB)/LB-agar medium (Sambrook et al.

The results demonstrate the potential of GFP labeling for protein

The results demonstrate the potential of GFP labeling for protein functional characterizations in Xac, and, in addition, the Xac mutant strain labeled at the septum constitutes a biological model for the exploration of antibacterial compounds able to inhibit cell division in this plant pathogen. Xanthomonas

citri ssp. citri (Schaad et al., 2005, 2006) (also known as Xanthomonas axonopodis pv. citri or Xac) is a Gram-negative, plant-pathogenic bacterium that affects most citrus species and is the causal agent of citrus canker, a very economically important disease of citrus plants worldwide. An effective control for this disease is inexistent, and a more detailed understanding of the biology of the etiological agent may contribute substantially toward the development of strategies to prevent and control infection. A major effort to accomplish selleckchem this task was the elucidation of the genome sequence of Xac (da Silva et al., 2002), which has stimulated a number of molecular studies using Xac as model microorganism,

and yet, little information is available regarding technical methods that could enhance its proteome exploration (Galvao-Botton et al., 2003; Mehta & Rosato, 2003; Alegria et al., 2004, 2005; Cernadas et al., 2008). Our main interest focused on the characterization of some essential biological processes of Xac, more specifically those PS-341 supplier involved with chromosome segregation and cell division. A common feature of such bacterial systems is that they are usually composed of proteins sharing little homology to their functional analogues in more derived eukaryotes; therefore, these proteins constitute ideal targets for antimicrobial drug development and pathogen control (e.g. Gitai et al., 2005; Pan et al., 2006; Haydon et al., Selleck Sunitinib 2008; Beuria et al., 2009; Kapoor & Panda, 2009). However, to undertake protein functional studies with/in Xac, we were limited

by the lack of biological tools developed and/or accessible for this purpose. Here, we describe a protein expression system dedicated to Xac, which can also be used for the subcellular localization of the green fluorescent protein (GFP)-labeled factors in this pathogen. We used the system to characterize a hypothetical protein of Xac that shares significant homology to the FtsZ-stabilizing factor ZapA, originally described in Bacillus subtilis (ZapABsu) (Gueiros-Filho & Losick, 2002). Furthermore, we show that the disruption of the α-amylase gene, the site of plasmid integration into the Xac chromosome, does not alter its pathogenesis. The Xanthomonas citri ssp. citri used was the sequenced strain (da Silva et al., 2002), formerly designated X. axonopodis pv. citri strain 306 (IBSBF 1594). Escherichia coli strain DH10B (Invitrogen) was used for cloning. Escherichia coli was cultivated at 37 °C in a Luria–Bertani (LB)/LB-agar medium (Sambrook et al.