Furthermore, as all sequence reads were delivered in FASTA format (SRA050786), the present study also provides a substantial genomic basis for A. subrufescens and, more generally, contributes to basidiomycete resources. Our results confirmed that the main limitation to SSR marker development now lies with the effectiveness of screening tests for marker validation rather than on the isolation process (Gardner et al., 2011; Malausa et al., 2011). This was particularly relevant in fungal species for which polymorphism was lower than in other phylogroups (Dutech et al., 2007). We have demonstrated the usefulness of the present set of microsatellite loci for A. subrufescens

for subsequent genetic diversity, fingerprinting and mapping analyses. This also may provide a valuable tool to resolve the taxonomic controversy associated with this species (Wasser et al., 2002, 2005; Kerrigan, 2005, 2007; Wisitrassameewong et al., 2012). In addition, www.selleckchem.com/products/obeticholic-acid.html the transferability to closely related species offers opportunities to study speciation process and phylogenetic relationships within the Agaricus section Arvenses.

This research is a part of a research project funded by a bilateral cooperation between Mexico (project 115790 CONACYT) and France (project ‘AgaSub’ AG-014699 clinical trial ANR-09-BLAN-0391). We would like to thank Sylvie Richard-Cervera for her technical assistance in microsatellite genotyping. We gratefully thank D. Royse, L.A. Parra, M. Verfaillie, D.C. Zied, R.L. Zhao, and all the other mycologists for providing useful fungal material. Special thanks go to R.W. Kerrigan who provided strains and also helpful suggestions to improve this manuscript. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“In Saccharomyces cerevisiae, genes involved in thiamin pyrophosphate (TPP) synthesis (THI genes) and the pyruvate decarboxylase structural gene PDC5 are transcriptionally induced in response

to thiamin starvation. Three positive regulatory factors (Thi2p, Thi3p, and Pdc2p) are involved in the expression of THI genes, whereas only Pdc2p is required for the expression of PDC5. Thi2p and Pdc2p serve as transcriptional activators Casein kinase 1 and each factor can interact with Thi3p. The target consensus DNA sequence of Thi2p has been deduced. When TPP is not bound to Thi3p, the interactions between the regulatory factors are increased and THI gene expression is upregulated. In this study, we demonstrated that Pdc2p interacts with the upstream region of THI genes and PDC5. The association of Pdc2p or Thi2p with THI gene promoters was enhanced by thiamin starvation, suggesting that Pdc2p and Thi2p assist each other in their recruitment to the THI promoters via interaction with Thi3p.

As DAPT solubility dmso a result the method was adapted such that different amounts of RNA (10, 20, 50, 100, and 150 ng of the normally used 200 ng RNA) were used in the reverse transcription reaction. Subsequently identical volumes of these reactions were used as template in real-time experiments. The standard curves for the three genes used (Uf-CON1, Uf-CON2, and Uf-TBB1) are depicted in Fig. 2a. The slopes of the three standard curves are almost identical. However, the standard curve for Uf-TBB1 is markedly shifted to higher Ct values, reflecting lower levels of transcript abundance of Uf-TBB1 compared with

the two other genes (Uf-CON1 and Uf-CON2). For the quantification of haustoria, three genes (Uf-HXT1, Uf-RTP1, and Uf-THI1) were used, which have been shown to be haustorium-specifically expressed (Hahn & Mendgen, 1997; Voegele et al., 2001). Again slopes of the standard curves are almost identical (Fig. 2b). The low CT numbers indicate high levels of transcript abundance. Indeed, all three genes have been shown to be among the most highly expressed genes in haustoria, representing between 0.7% and 2.8% of the total cDNA each (Hahn & Mendgen, 1997; Voegele et al., 2001). These standard curves were then used to perform an absolute quantification of U. fabae in planta. Figure 3a–c depicts the fraction of the constitutively expressed genes Uf-CON1 (a), Uf-CON2

(b), and Uf-TBB1(c) of the total RNA of samples from infected leaves as a function of

disease progression. These results mirror those obtained with dot plot analysis. It appears that there is a lag phase in HDAC inhibitor mechanism the early days after inoculation, where hardly any fungus is detectable. Between 4 and 8 dpi, there is an exponential increase of the proportion of RNA made up by the fungus. Thereafter, the fungal fraction seemed to reach a steady-state level of around 50% of the total RNA. Results from these analyses correlated so well that data for the different genes could be integrated into a single graph (Fig. 4a). The fact that the proportion of fungal RNA does not seem to increase continuously might reflect the specific need of obligate biotrophic pathogens PAK6 to keep their host plants alive in order to assure propagation. Nine days post inoculation an equilibrium seems to be established enabling further pathogen development and proliferation without damaging the host plant to a point where it ceases growth. The proportion of about 50% fungal RNA is considerably higher than the amount of 20% fungal DNA reported for a compatible interaction of the poplar rust Melampsora medusae with its host (Boyle et al., 2005). This discrepancy might either be due to the problems associated with using DNA for quantification of rust fungi mentioned above, or to different levels of pathogen present in different host–parasite interactions. Jakupovic et al.

1.2 We recommend EFV in combination with TDF and FTC as first-line ART in TB/HIV coinfection. 1C   We recommend that when rifampicin is used with EFV in patients over 60 kg, the EFV dose is increased to 800 mg daily. Standard Doramapimod clinical trial doses of EFV are recommended

if the patient weighs <60 kg. 1C   We recommend that rifampicin is not used with either NVP or a PI/r. 1C   We recommend that where effective ART necessitates the use of PI/r that rifabutin is used instead of rifampicin. 1C CD4 cell count (cells/μL) HBV requiring treatmenta HBV not requiring treatment HCV with immediate plan to start HCV treatmenta HCV with no immediate plan to start HCV treatment a See BHIVA Guidelines for the management of coinfection with HIV-1 and hepatitis B or C virus [1] for indications CHIR99021 to treat hepatitis B and C. Start ART in some patients (2C) (Include TDF and FTC) Start ART (1B) (Include TDF and FTC) Start ART (1B) (Include TDF and FTC) 350–500 Start ART after HCV treatment commenced (1C) <350 Start ART before HCV treatment (1B) Discuss with HIV and viral hepatitis specialist 8.2.2.1 ● We

recommend patients with HIV and hepatitis B virus coinfection who have a CD4 cell count <500 cells/μL are treated with fully suppressive ART inclusive of anti-HBV active antivirals. 1B   ● We recommend patients with HIV and HBV coinfection who have a CD4 cell count ≥500 cells/μL and who have an HBV-DNA ≥2000 IU/mL and/or evidence of more than minimal fibrosis (Metavir ≥F2) are treated with fully suppressive ART inclusive of anti-HBV active antivirals. 1C 8.2.2.2 ● We recommend

TDF/FTC as part of a fully suppressive ART combination should be given to all patients where HBV treatment is deemed necessary. 1C   ● We recommend neither 3TC nor FTC be used as the sole active drug against HBV in ART due to the rapid emergence of HBV resistant to these agents. 1B   ● We recommend 3TC/FTC may be omitted from the ART regimen and tenofovir be given ADP ribosylation factor as the sole anti-HBV active agent if there is clinical or genotypic evidence of 3TC/FTC-resistant HBV or HIV. 1D 8.2.3.1 ● We recommend all patients with HIV and hepatitis C virus coinfection be assessed for HCV treatment. GPP   ● We suggest commencing ART when the CD4 cell count is greater than 500 cells/μL in all patients who are not to commence HCV treatment immediately. 2D   ● We recommend commencing ART when the CD4 cell count is less than 500 cells/μL in all patients who are not to commence anti-HCV treatment immediately. 1B   ● We recommend commencing ART to optimize immune status before anti-HCV therapy is initiated when the CD4 cell count is between 350 and 500 cells/μL unless there is an urgent indication for anti-HCV treatment when ART should be commenced as soon as the patient has been stabilized on HCV therapy.

Travel destinations were grouped as follows: Africa, Asia, Europe, Latin America/Caribbean, and the USA. A “Multiple/other” category was used for cases that traveled to more than one destination group or that traveled to other parts of the world (eg, Australia). INK 128 manufacturer The data variables used for the analysis included age; gender; onset date; disease; symptoms

(abdominal pain, abdominal bloating, chills, dehydration, diarrhea, bloody diarrhea, greasy diarrhea, chronic diarrhea, dizziness, fatigue, fever, headache, loss of appetite, weight loss, muscle soreness, nausea, vomiting, and weakness, depending on the disease); hospitalization; recovery date; travel destination; resort accommodation; mode of transportation; and departure and return dates. Where data were available, disease duration was computed as recovery

minus onset dates, and travel duration as return minus departure dates. The TRC were broken down by illness and then further described by hospitalization, symptoms, disease duration, age, gender, travel duration, travel destination, mode of transportation, and resort accommodation. When available, onset, travel departure, and return dates were plotted by month. For each of these three dates, a Poisson regression model of the monthly count was used to test the differences LDK378 between both years and months at the same time. The year was defined as a successive 12-month period starting from June and ending in May the following year; eg, 2005 to 2006 encompassed June 2005 to May 2006 Pazopanib datasheet inclusively.

Multiple correspondence analysis (MCA) was used to probe the existence of travelers’ subgroups within the data and, if these existed, to explore any association between these subgroups and illness. MCA is a descriptive statistical technique designed to explore and visualize the relationships between three or more categorical variables (see Appendix 1 for more details).23 MCA was conducted on age (<5, 5–14, 15–24, 25–39, 40–59, and 60+ y), gender, travel duration (<8, 8–28, and 29+ d), travel destination, and accommodation in resort. In addition, disease was the supplementary variable used to assess any relationship between the subgroups identified and the illnesses. Finally, TRC and DC were overall described by, and tested for differences in, gender, age, illness distribution, and hospitalization. For every disease with at least 30 TRC, TRC and DC were compared for gender, age, hospitalization, disease duration, the various symptoms relevant to the disease, and the delay of reporting (difference between onset and report dates). Campylobacter species and Salmonella serotypes were also compared. The Kruskal–Wallis test was used for continuous data and the bilateral Fisher exact test or the Chi-square test for categorical data. The p-value threshold was set at 0.01 for all statistical tests.

The peak amplitude of RON was larger to voice as compared to music deviants over Small molecule library midline (F1,34 = 8.78, P < 0.01, ηp2 = 0.205) and mid-lateral (F1,34 = 7.508,

P = 0.01, ηp2 = 0.181) sites, with a trend in the same direction over lateral sites (F1,34 = 3.102, P = 0.087, ηp2 = 0.084), pointing to a greater ease at overcoming distraction when deviants were vocal as compared with musical in nature. Lastly, the mean amplitude of RON was significantly larger over the right as compared with the left hemisphere in lateral sites (F1,34 = 21.238, P < 0.01, ηp2 = 0.384), with a trend in the same direction in mid-lateral sites (F1,34 = 3.683, P = 0.063, ηp2 = 0.098). To determine whether an enhanced N1 is correlated with behavioral measures Selleckchem Everolimus of musical expertise, we examined a connection between the N1 peak amplitude and the following measures: onset of musical training, years of musical training, MAP scores, self-rated musical proficiency and the number of hours listening to music per week. The N1 peak amplitude averages were calculated for midline, mid-lateral and lateral sites for

each participant. Separate regression analyses were performed between each of the above behavioral measures and the N1 average for each scalp area. Because the amplitude of N1 was significantly smaller in response to standards compared with deviants [probably due to the refractoriness of the neurons responding to repeating standard sounds (Näätänen & Picton, 1987)], we conducted separate regression analyses on N1 to standards and on N1 to deviants. All reported P values are two-tailed. In the NAT condition, individuals with higher self-rated musical proficiency had a significantly larger N1 peak amplitude to both music and voice deviants over the mid-lateral (music deviants, r = 0.371, ADAMTS5 P = 0.026; voice deviants, r = 0.338, P = 0.044) and midline (music deviants, r = 0.351, P = 0.036; voice deviants, r = 0.342, P = 0.041) sites, with a trend in the same direction over the lateral sites (music deviants, r = 0.315, P = 0.061; voice deviants, r = 0.281, P = 0.097).

Additionally, the N1 elicited by music deviants was larger over the lateral sites (r = 0.357, P = 0.032) in individuals with higher MAP scores. A relationship between the N1 peak amplitude to voice deviants and MAP scores showed a similar trend (r = 0.291, P = 0.085). None of the results in the ROT condition reached significance. In the NAT condition, individuals with higher self-rated musical proficiency had a significantly larger N1 peak amplitude to both music and voice standards over the midline (music standards, r = 0.335, P = 0.046; voice standards, r = 0.402, P = 0.015) and mid-lateral (music standards, r = 0.331, P = 0.049; voice standards r = 0.385, P = 0.02) sites. Individuals with higher MAP scores had a larger N1 to voice standards (r = 0.342, P = 0.041) and a marginally larger N1 to music standards (r = 0.295, P = 0.081) over the lateral sites.

0002 μmol N2O L−1). However, after 10 days, the O2 concentrations had declined to a mean of 5.6% v/v in all the treatments (Fig. 2a) due to O2 dissolution. N2O concentrations at day 10 in the Protein Tyrosine Kinase inhibitor headspace of the three fungal treatments were 0.0117±0.00015 μmol N2O L−1 (P. involutus), 0.0114±0.0003 μmol N2O L−1 (T. fibrillosa) and 0.0114±0.00043 μmol N2O L−1 (F. lichenicola); there was no difference in the headspace N2O concentrations between the three species. No N2O was detected in the control flasks, which indicates a fungal source for N2O production. N2O production can contribute to cell growth (e.g. Shoun

& Tanimoto, 1991; Zhou et al., 2001); however, the selleck compound energy yield varies between species (Usuda et al., 1995), and further work is required to determine whether N2O production is an energy-yielding process for symbiotic ectomycorrhizal fungi compared with free-living fungi. The CO2 concentrations increased in all the fungal treatments (Fig. 2b), and were significantly higher (P<0.05) in the ectomycorrhizal fungal treatments by day 10. There was a significant decline in the nitrate concentrations over 10 days for P. involutus (Fig. 2c),

resulting in the recovery of 0.006% of the original medium nitrate-N as N2O-N by day 10. Although the final media pH differed significantly between treatments (Fig. 2d), there was no significant change over the incubation period within treatments; hence, it is unlikely that N2O detection can be attributed to abiotic production from nitrite. Caution must be exercised when making direct comparisons with data from other studies, due to the very different culture conditions and growth periods, as our results are below the range of N2O fluxes published thus far, which may range from <100 to 1000 μmol N2O over shorter growth periods PIK-5 than presented here. For example, the maximum N2O production by F.

lichenicola was ∼600 μmol N2O [20 mM NaNO3 with ammonium after 48 h (Fig. 1b; Watsuji et al., 2003)] and F. oxysporum produced ∼800 μmol N2O [after 96 h; 10 mM NaNO3 (Fig. 1a; Shoun & Tanimoto, 1991)]. Further investigation is required to determine ectomycorrhizal fungal N2O production under a wider range of O2, N and C conditions. In our experiment, N2O production was detected when the O2 concentrations were <6% v/v O2 (day 10), suggesting that the two ectomycorrhizal fungi we examined here may possess the ability to reduce nitrate as an alternative respiratory mechanism. This may be of important environmental relevance in situations where CO2 concentrations are particularly high due to, for example, increased microbial activity within the fungal hyphosphere (e.g. Baschien et al., 2009).

When E. coli was used as donor, no transfer of pKJK10 was detected to any of the individual 15 soil isolates, but P. putida was observed to transfer pKJK10 to Stenotrophomonas rhizophila. The plasmid transfer frequency from P. putida buy VX-770 to S. rhizophila

was higher when the filters were placed on TSA medium (1.07 ± 3.05 × 10−1) compared with R2A medium (0.33 ± 2.32 × 10−2, Table 2), supporting the fact that the metabolic state of the cells may in some cases influence conjugation frequencies (van Elsas & Bailey, 2002). These results reflect the fact that the host range of plasmids depends on the identity of the donor strain (De Gelder et al., 2005). 1.07 ± 3.05 × 10−1 0.33 ± 2.32 × 10−2c In contrast to the results observed when transferring pKJK10 to individual isolates, no plasmid transfer events were observed from P. putida to the mixed community consisting of the same 15 strains applied individually above. Transconjugants were, however, obtained when applying E. coli as donor of pKJK10. The green fluorescent transconjugant cells were sorted by FACS and cultured on TSA agar plates. By sequence analysis of the 16S rRNA gene from four colonies from each replicate, the selected transconjugants were shown all to be identical and identified

as Ochrobactrum rhizosphaerae. YAP-TEAD Inhibitor 1 clinical trial This does not exclude the possibility that other isolates may also have received the plasmid, but it does show that O. rhizosphaerae

in fact did so and that it was the most dominant strain among the Dynein plasmid recipients. Interestingly, O. rhizosphaerae was not able to receive the plasmid in the individual mating experiment, indicating that the plasmid permissibility does not only depend on the abilities of the plasmid, host and recipient strains, but also on the surrounding microbial community, which may reduce or enhance plasmid transfer. Both of these scenarios were observed in this study; transfer of pKJK10 from P. putida to S. rhizophila was observed in diparental mating experiments, but not in a mixed community, possibly caused by reduced survival/competition ability of the strains or by the fact that the donor and this specific recipient populations had less opportunity for interaction in the mixed community. In contrast, the presence of a mixed community induced pKJK10 transfer from E. coli to O. rhizosphaerae, which may be due to altered physical cell–cell interaction or the presence of one or several intermediate plasmid host(s). These ‘plasmid step-stones’ may facilitate plasmid transfer from E. coli to O. rhizosphaerae, but are unable to establish and stabilize the plasmid in their own population. Because it was not possible to isolate the strains individually after growth in the community, the fraction of O. rhizosphaerae herein could not be determined; It is possible that O.

Such effects of porin alterations on cephalosporin resistance levels in β-lactamase-producing enterobacteria have been well documented (Martínez-Martínez, 2008). In the other pair of isolates of the PFGE subtype B1, C-S isolate P2/I177971 and C-NS isolate P2/I168905, a general increase in β-lactam MICs was also observed. However, it had another nature and there were also significant differences across these two related pairs of isolates, namely between the two C-S isolates (P3/C154247 and P2/I177971) and the two C-NS isolates (P3/A18867 and P2/I168905) in the levels of resistance

to different β-lactams. These observations suggest that other unidentified mechanisms have been accumulating in particular K. pneumoniae strain variants as it was also indicated in other reports (Gröbner Selleckchem HM781-36B et al., 2009). This work PD0325901 clinical trial contributes to the growing number of reports on C-NS Enterobacteriaceae strains due to ESBL and/or AmpC expression combined with porin alterations (Livermore & Woodford, 2006; Lee et al., 2007; Martínez-Martínez, 2008; Gröbner et al., 2009; Wang et al., 2009). Despite the recent dissemination of organisms with various types of carbapenemases, this mechanism remains an important

source of resistance to carbapenems in enterobacteria. The study reported here was financed by the research project grant MSMT 2E08003 from the Ministry of Education, and the project grant NS9717-4/2008 from the Ministry of Health, Czech Republic. The authors would like to thank to V.J. Benedí for kindly providing the polyclonal antibodies

against OmpK35 and OmpK36 porins. “
“Pseudomonas fluorescens 2P24 is an effective biological control agent of a number of soilborne plant diseases caused by pathogenic microorganisms. Among a range of secondary metabolites produced by strain 2P24, the antibiotic 2,4-diacetylphloroglucinol (2,4-DAPG) is the major determinant of its disease-suppressive capacity. In this study, we performed random mutagenesis using mini-Tn5 in order to screen for the transcriptional regulators of the phlA gene, a biosynthase gene responsible for 2,4-DAPG production. The mutant PMphlA23 with significantly decreased phlA gene expression was identified from ∼10 000 insertion colonies. The protein Metalloexopeptidase sequence of the interrupted gene has 84% identity to Hfq, a key regulator important for stress resistance and virulence in Pseudomonas aeruginosa. Genetic inactivation of hfq resulted in decreased expression of phlA and reduced production of 2,4-DAPG. Furthermore, the hfq gene was also required for the expression of pcoI, a synthase gene for the LuxI-type quorum-sensing signaling molecule N-acyl-homoserine lactone. Additionally, the hfq mutation drastically reduced biofilm formation and impaired the colonization ability of strain 2P24 on wheat rhizospheres. Based on these results, we propose that Hfq functions as an important regulatory element in the complex network controlling environmental adaption in P. fluorescens 2P24.

, 2009). We first noticed that the yicJI mutant formed smaller colonies than the wild type and the Δfrz strains on LB-agar plates. We then compared the growth of the wild-type strain, the Δfrz mutant, and the ΔJI mutant during MDV3100 order agitated and static cultures in LB-medium. Whereas the growth curves of the wild-type and of the Δfrz mutants were similar under both conditions,

the ΔJI mutant was affected in its ability of adaptation to the stationary phase of growth (OD600 nm of the ΔIJ mutant culture is 1 or 0.7 U lower than that of the wild-type strain after 72 h of agitated or static growth, respectively; Fig. 4). We reported previously that the frz operon is involved in the survival mechanism of BEN2908 during the late stationary growth phase in LB medium and in serum. Indeed, during co-cultures under oxygen-restricted conditions (static cultures), the wild-type strain BEN2908 outcompeted Anti-infection Compound Library purchase the BEN2908Δfrz strain during the late stationary growth phase, but not during the

exponential growth phase. This phenotype is strongly affected by oxygenation, as it is not revealed when the co-cultures are agitated (Rouquet et al., 2009). We thus tested the survival ability of the ΔJI mutant under these co-culture conditions, and we found that its fitness is strongly affected during the late stationary phase of growth, even when the co-cultures are highly agitated (Fig. 5, A–C). As the effect of the Frz system on the survival ability of the bacteria during the late stationary phase of growth was found to depend on the composition

of the culture medium, we analyzed the survival ability of the ΔJI mutant during static co-cultures with the wild-type strains in minimal media in which the fitness of the Δfrz mutant is not or only slightly affected (d-glucose, d-fructose, d-sorbose, and d-psicose). In contrast to the Δfrz mutant, the survival ability of the ΔJI mutant is strongly affected during the late stationary phase of growth in all these minimal media (Fig. 5, D–G). As isoprimeverose was found to be a substrate of YicI, we also tested the survival ability of the ΔJI and the Δfrz mutants during static co-cultures Ergoloid with the wild-type strain in a minimal medium containing this sugar as a sole carbon source. Again, the fitness of the ΔJI mutant was strongly affected during the late stationary phase of growth (6.2 ± 1.0% of mutant in the population after 7 days of co-culture), whereas that of the Δfrz mutant was not (53.7 ± 1.6% of mutant in the population after 7 days of co-culture). In conclusion, although the phenotypes of the Δfrz and the ΔJI mutants are not completely similar, both frz and yicJI metabolic operons are involved in the fitness of the bacteria and are cotranscribed through molecular mechanisms that could involve the FrzR activator and phosphoryl group transfer.

Other variables assessed included antiretroviral treatment experience and HIV practice size. To determine antiretroviral treatment experience prior to target medication initiation, we identified veterans who had received any VHA nontarget antiretroviral medication >7 days prior to their first prescription of the target medication. Those with no such prior antiretroviral prescriptions were defined as antiretroviral naïve. We compared the proportion of veterans who were antiretroviral naïve who started on each target medication in the first two quarters post-approval with the proportion who were antiretroviral naïve who started on each target medication in the subsequent quarters post-approval. HIV practice size was

categorized as small (≤100 patients), medium (101–300), large (301–600) and very large (>600) based http://www.selleckchem.com/products/EX-527.html on the mean number of HIV-positive patients in care in each quarter from 1 April 2003 until 31 December 2007. χ2 tests were used to evaluate differences in target medication uptake by period and between regions. Data were analysed using sas version 8.2 (SAS Institute, Cary, NC, USA). This protocol was approved by the VA Palo Alto Health Care System Office of Research Administration, the Stanford University Institutional Review Board

and the VHA Clinical Case Registry Research Committee. Data are presented for 128 distinct reporting Selleckchem Staurosporine VHA facilities with a median of 141 HIV-infected veterans in care per year until per facility between 2003 and 2007 (range 1–1147). Geographically, there were 27 facilities in the Northcentral, 23 in the Northeast, 48 in the South, and 30 in the West. In any quarter, there were between 3500 and 4000 providers who prescribed antiretrovirals. During the study period, 5667 HIV-infected veterans started atazanavir (5618 through 18 complete quarters), 559 started darunavir (542 through

six quarters), 325 started tipranavir (322 through 10 quarters) and 7844 started lopinavir/ritonavir (5927 through 18 quarters). The number of new prescriptions for each target medication in each quarter post-approval is shown in Figure 1. The number of new prescriptions for atazanavir per quarter was generally consistent (approximately 400 prescriptions per quarter) until the tenth quarter post-approval when uptake began to decline steadily. Atazanavir uptake closely mimicked the uptake pattern of lopinavir/ritonavir. Darunavir uptake remained steady across the early quarters until the sixth quarter when new prescriptions substantially increased. Tipranavir uptake declined considerably after the second quarter post-approval; new prescriptions decreased from the seventies to the teens within three quarters. This antiretroviral had the largest decrease in uptake over time. In terms of provider type, for all medications, in period 1 the majority of new prescriptions were written by physicians with approximately 20–30% written by physician extenders (Fig. 2).