, 2007). Ohki & Tateno (2004) described the increased expression of the bmr3 efflux transporter due to a double mutation at positions −18 and +4 from the transcription start site. Transcriptional lacZ reporter gene fusions with a region upstream of the bmrA SD sequence were constructed and integrated by double crossing

over into the amyE locus of the B. subtilis 168 chromosome. Measurements of β-galactosidase activity determined the putative promoter region (Fig. 2). Subsequently, primer extension was used to identify the transcription start downstream of a potential promoter (Fig. 2a). The wild-type promoter shows a nearly perfect −10 box with TATGAT, a 17-bp spacer, but a weak −35 box with CTGAAA. In mutant 8R, the C of the −35 box was altered to T, making it more similar to the consensus σA−35 box TTGACA (Fig. 2b). The second point mutation was located six bases downstream from the transcription start site (+6) altering PD0325901 supplier an A5 stretch to GAAAA (Fig. 1b). To dissect

the contribution of each single mutation on the elevated expression of bmrA, plasmids carrying transcriptional bmrA–lacZ fusions with fragments of different sizes were constructed designated pACMM (double mutant), pACWW (wild type), pACMW (−35 mutation) and pAWM (+6 mutation) (Fig. 1a). All pAC6 derivatives were integrated into the amyE locus PARP inhibition and the β-galactosidase activities measured (see Fig. 1a). Increased β-galactosidase activities compared with the wild type were found in the double mutant and in the single mutant affecting the −35 box, whereas only marginally different β-galactosidase activities were measured for the +6 mutation (3.5-fold increased). The 157-bp upstream region increased β-galactosidase 10–11-fold in both the double and the MW mutant compared with the wild type. To investigate the impact of the mutations

on bmrA http://www.selleck.co.jp/products/Abiraterone.html expression, total RNA from wild-type strain 168 and mutant 8R was isolated, DNase treated and assayed using real-time PCR. The amount of bmrA mRNA in mutant 8R with the −35 and +6 mutations was 135-fold increased, whereas in strain YH2M with the −35 mutation alone, the amount of bmrA mRNA was about 13-fold increased (Fig. 2(b). 8R-ind). Real-time PCR on total RNA isolated from B. subtilis 8R propagated in the presence of CmC (0.5 μM) corroborated the results of the Jault laboratory on the constitutive expression of bmrA (Steinfels et al., 2004). To analyze the binding of the RNAP to the bmrA promoter region, EMSAs were performed. The four 157-bp fragments used for the lacZ-reporter gene fusions were radioactively labelled and incubated with increasing concentrations of B. subtilis RNAP. As shown in Fig. 3a and b, the −35/+6 mutant MM and the single −35 mutant MW displayed a 30-fold increased affinity for RNAP. Interestingly, the single mutant WM carrying only the +6 mutation behaved like the wild type. The addition of heparin (Fig.

S1a), as described under ‘Materials and methods’. Topology models predicted that the N-terminal end of B. subtilis Chr3N was located in the periplasm, just about 12 residues Crenolanib chemical structure distal of TMS1 (Fig. S1b). Fusions were not constructed in this short hydrophilic region because Chr3N-PhoA recombinant proteins would remain in the cytoplasm by lacking a TMS that might translocate PhoA to the periplasm. The shortest Chr3N fusion, made in residue Gly24 (predicted to reside within TMS1, close to the cytoplasm), yielded high LacZ activity and no significant PhoA activity (Fig. 1a). Thus, the presence of TMS1 could not be clearly demonstrated, and we rely on the prediction of the topology models

to suggest that the N-terminal end of Chr3N is located in the periplasmic space (Fig. S1b). Fusions located in amino acids Asn37, Ile50, and Lys74 showed LacZ activity and null PhoA activity (Fig. 1a), indicating that this

region is situated in the cytoplasm; this location is in agreement with prediction models (Fig. S1b), which showed large hydrophilic (cytoplasmic) regions between residues 50 and 90. Fusions at residues His106, Leu137, Ile161, and Ser189 yielded alternating high and low PhoA activities (Fig. 1a), indicating that these regions have corresponding alternate periplasmic and cytoplasmic locations; this location was confirmed by the selleck products fact that these four fusions also yielded alternating low and high LacZ activities (Fig. 1a). The topology at this region, which spans the last four TMSs of Chr3N, is in complete agreement with prediction models (Fig. S1b). Together, these results suggested a topology of five TMSs for Chr3N, with the N-terminal end in the periplasm and the C-terminal end in the cytoplasm (Fig. 1b). Topology

models predicted that the N-terminal end of B. subtilis Chr3C was located in the cytoplasm (Fig. S1b). Accordingly, fusions located in amino acids Tyr36 and Met47 showed both high PhoA activity and low LacZ activity (Fig. 1c), indicating that this region was situated in the periplasm; a TMS should be present distal of Tyr36 to allow for this region to be translocated to the periplasm and to yield PhoA enzyme activity. These data confirmed that the N-terminal of Chr3C is located Chloroambucil in the cytoplasm. Topology models predicted a large hydrophilic (periplasmic) Chr3C region spanning residues 50 through 90 (Fig. S1b). However, fusions at Val66 and Ala70 displayed unexpectedly low and null PhoA activity, respectively (Fig. 1c); the Ala70 fusion showed low LacZ activity, indicating that it was not at the cytoplasm. As fusion at Gly109 showed significant LacZ activity, a TMS must be present between residues 70 and 109, as predicted (Fig. S1b); this means that the 66–70 upstream region must be located in the periplasm.

Clinical and virological outcomes in HIV-infected patients with chronic hepatitis B on long-term nucleos(t)ide analogues. AIDS 2011; 25: 73–79. 31  Puoti M, Cozzi-Lepri A, Arici C et al. Impact of lamivudine on the risk of liver related death in 2,041 HBsAg- and HIV-positive

individuals: results from an inter-cohort analysis. Antivir Ther 2006; 11: 567–574. 32  Marra F, Bruno R, Galastri S. gp120 induces directional migration of human hepatic stellate cells: a link between HIV infection and liver fibrogenesis. Hepatology 2007; 46: Abstract A125. 33  Tuyama AC, Hong F, Saiman Y et al. Human immunodeficiency virus (HIV)-1 infects human hepatic stellate cells and promotes collagen I and monocyte chemoattractant protein-1 expression: implications for the pathogenesis of HIV/hepatitis C virus–induced liver fibrosis. Hepatology 2010; 52: PLX4032 nmr 612–622. 34  Marchetti G, Tincati C, Silvestri G. Microbial translocation in the pathogenesis

of HIV infection and AIDS. Clin Microbiol Rev 2013; 26: 2–18. 35  Aoyama T, Paik YH, Seki E. Toll-like receptor signaling and liver fibrosis. Gastroenterol Res Pract 2010; Article ID 192543, 8 pages. 36  Yuen MF, Yuan HJ, Wong D et al. Prognostic determinants for chronic hepatitis B in Asians: therapeutic implications. Gut 2005; 54: 1610–1614. 37  Yuan HJ, Yuen MF, Wong D, Sablon E, Lai CL. The relationship between HBV-DNA levels and cirrhosis-related complications in Chinese Progesterone selleck compound with chronic hepatitis B. J Viral Hepat 2005; 12: 373–379. 38  Iloeje UH, Yang HI, Su J, Jen CL, You SL, Chen CJ for the Risk Evaluation of Viral Load Elevation and Associated Liver Disease/Cancer-In HBV (the REVEAL-HBV) Study Group. Predicting cirrhosis risk based on the level of circulating hepatitis B viral load. Gastroenterology 2006; 130: 678–686. 39  Gane EJ, Lim TH, Moyes C, Cunningham

C. 71 predictors of liver complications in childhood-acquired HBV infection in New Zealand Maori: results of 27 year longitudinal study. J Hepatol 2012; 56(Suppl 2): S31. 40  Liaw YF. Impact of therapy on the outcome of chronic hepatitis B. Liver Int 2013; 33(Suppl 1): 111–115. 41  Lacombe K, Rockstroh J. HIV and viral hepatitis coinfections: advances and challenges. Gut 2012; 61(Suppl 1): i47–i58. 42  Di Martino V, Thevenot T, Colin JF et al. Influence of HIV Infection on the response to interferon therapy and the long-term outcome of chronic hepatitis B. Gastroenterology 2002; 123: 1812–1822. 43  Johnson RM, Ristig MB, Overton ET, Lisker-Melman M, Cummings OW, Aberg JA. Safety and tolerability of sequential pegylated IFN-α2a and tenofovir for hepatitis B infection in HIV+ individuals. HIV Clin Trials 2007; 8: 173–181. 44  European Association for the Study of the Liver. EASL Clinical Practice Guidelines: Management of chronic hepatitis B virus infection. J Hepatol 2012; 57: 167–185. 45  Benhamou Y, Bochet M, Thibault V et al.

Clinical and virological outcomes in HIV-infected patients with chronic hepatitis B on long-term nucleos(t)ide analogues. AIDS 2011; 25: 73–79. 31  Puoti M, Cozzi-Lepri A, Arici C et al. Impact of lamivudine on the risk of liver related death in 2,041 HBsAg- and HIV-positive

individuals: results from an inter-cohort analysis. Antivir Ther 2006; 11: 567–574. 32  Marra F, Bruno R, Galastri S. gp120 induces directional migration of human hepatic stellate cells: a link between HIV infection and liver fibrogenesis. Hepatology 2007; 46: Abstract A125. 33  Tuyama AC, Hong F, Saiman Y et al. Human immunodeficiency virus (HIV)-1 infects human hepatic stellate cells and promotes collagen I and monocyte chemoattractant protein-1 expression: implications for the pathogenesis of HIV/hepatitis C virus–induced liver fibrosis. Hepatology 2010; 52: Silmitasertib cost 612–622. 34  Marchetti G, Tincati C, Silvestri G. Microbial translocation in the pathogenesis

of HIV infection and AIDS. Clin Microbiol Rev 2013; 26: 2–18. 35  Aoyama T, Paik YH, Seki E. Toll-like receptor signaling and liver fibrosis. Gastroenterol Res Pract 2010; Article ID 192543, 8 pages. 36  Yuen MF, Yuan HJ, Wong D et al. Prognostic determinants for chronic hepatitis B in Asians: therapeutic implications. Gut 2005; 54: 1610–1614. 37  Yuan HJ, Yuen MF, Wong D, Sablon E, Lai CL. The relationship between HBV-DNA levels and cirrhosis-related complications in Chinese 4-Aminobutyrate aminotransferase ABT-737 supplier with chronic hepatitis B. J Viral Hepat 2005; 12: 373–379. 38  Iloeje UH, Yang HI, Su J, Jen CL, You SL, Chen CJ for the Risk Evaluation of Viral Load Elevation and Associated Liver Disease/Cancer-In HBV (the REVEAL-HBV) Study Group. Predicting cirrhosis risk based on the level of circulating hepatitis B viral load. Gastroenterology 2006; 130: 678–686. 39  Gane EJ, Lim TH, Moyes C, Cunningham

C. 71 predictors of liver complications in childhood-acquired HBV infection in New Zealand Maori: results of 27 year longitudinal study. J Hepatol 2012; 56(Suppl 2): S31. 40  Liaw YF. Impact of therapy on the outcome of chronic hepatitis B. Liver Int 2013; 33(Suppl 1): 111–115. 41  Lacombe K, Rockstroh J. HIV and viral hepatitis coinfections: advances and challenges. Gut 2012; 61(Suppl 1): i47–i58. 42  Di Martino V, Thevenot T, Colin JF et al. Influence of HIV Infection on the response to interferon therapy and the long-term outcome of chronic hepatitis B. Gastroenterology 2002; 123: 1812–1822. 43  Johnson RM, Ristig MB, Overton ET, Lisker-Melman M, Cummings OW, Aberg JA. Safety and tolerability of sequential pegylated IFN-α2a and tenofovir for hepatitis B infection in HIV+ individuals. HIV Clin Trials 2007; 8: 173–181. 44  European Association for the Study of the Liver. EASL Clinical Practice Guidelines: Management of chronic hepatitis B virus infection. J Hepatol 2012; 57: 167–185. 45  Benhamou Y, Bochet M, Thibault V et al.

ananatis SC17(0). Deletion of the mentioned ORF (named gcd) from the P. ananatis SC17(0) genome led to the inability of mutant cells to accumulate gluconic acid in the media (Table 4) and to the abolition of GDH activity in their extracts (Table 2). Thus, it was confirmed that GU580893 indeed encoded GDH (likely membrane-bound GDH). Moreover, it could Pictilisib supplier be proposed that P. ananatis SC17(0) is able to oxidize glucose into gluconic acid by the fully active PQQ-mGDH

and that the corresponding genetic elements responsible for PQQ biosynthesis must be identified in the genome of this microorganism. A putative pqqABCDEF operon (GenBank accession number GU580892), structurally homologous to the similar genetic element in the Klebsiella pneumoniae chromosome (Meulenberg et al., 1992), was found in the genome of P. ananatis SC17(0) via a computer search. There was a high level of amino acid homology between putative polypeptides of P. ananatis and experimentally

confirmed proteins from K. pneumoniae: PqqB(84%), PqqC (91%), selleck inhibitor PqqD (75%), PqqE (84%) and PqqF(43%). For P. ananatis PqqA, differences were observed for two amino acid residues Thr6 and Val8. However, conservative Glu15 and Tyr19, which in the case of K. pneumoniae presumably appear as precursors of the PQQ molecule (Velterop et al., 1995), were at the same positions in the putative PqqA of P. ananatis. To determine whether the putative pqq operon was essential for PQQ biosynthesis in P. ananatis SC17(0), two types of strains were constructed; the

first lacked this genetic element and the second had an additional copy of the pqq operon in the chromosome. Deletion of the predicted P. EGFR antibody ananatis pqq operon led to the inability of mutant cells to accumulate gluconic (Table 4) acid and to the abolition of GDH activity in their extracts (Table 2) without exogenous PQQ in reaction in distinction from GDH extracted from P. ananatis SC17(0). Thus, it was confirmed that PQQ is indeed essential for the formation of active holoenzyme GDH and predicted pqq operon encoded genetic elements essential for PQQ biosynthesis. Construction of the strain SC17(0)-φ80attB-pqq was achieved by in vivo cloning of the pqq operon (see Supporting Information) and adaptation of ‘Dual In/Out’ Recombineering-driven strategy for the integration of DNA fragments into targeted points of the E. coli chromosome (Minaeva et al., 2008) for application in P. ananatis. The scheme applied in this work could be a useful instrument for the simple amplification of target genes/operons in the chromosome without preliminary amplification by PCR. The resulting strains, SC17(0)-Δpqq with the ΔpqqABCDEF operon and SC17(0)-φ80attB-pqq with two copies of this operon in the chromosome, were tested for their ability to accumulate PQQ in cultural medium. Inactivation of the putative pqq operon resulted in a decrease of PQQ in the medium to undetectable levels (<1 mg L−1).

ananatis SC17(0). Deletion of the mentioned ORF (named gcd) from the P. ananatis SC17(0) genome led to the inability of mutant cells to accumulate gluconic acid in the media (Table 4) and to the abolition of GDH activity in their extracts (Table 2). Thus, it was confirmed that GU580893 indeed encoded GDH (likely membrane-bound GDH). Moreover, it could 3-deazaneplanocin A manufacturer be proposed that P. ananatis SC17(0) is able to oxidize glucose into gluconic acid by the fully active PQQ-mGDH

and that the corresponding genetic elements responsible for PQQ biosynthesis must be identified in the genome of this microorganism. A putative pqqABCDEF operon (GenBank accession number GU580892), structurally homologous to the similar genetic element in the Klebsiella pneumoniae chromosome (Meulenberg et al., 1992), was found in the genome of P. ananatis SC17(0) via a computer search. There was a high level of amino acid homology between putative polypeptides of P. ananatis and experimentally

confirmed proteins from K. pneumoniae: PqqB(84%), PqqC (91%), see more PqqD (75%), PqqE (84%) and PqqF(43%). For P. ananatis PqqA, differences were observed for two amino acid residues Thr6 and Val8. However, conservative Glu15 and Tyr19, which in the case of K. pneumoniae presumably appear as precursors of the PQQ molecule (Velterop et al., 1995), were at the same positions in the putative PqqA of P. ananatis. To determine whether the putative pqq operon was essential for PQQ biosynthesis in P. ananatis SC17(0), two types of strains were constructed; the

first lacked this genetic element and the second had an additional copy of the pqq operon in the chromosome. Deletion of the predicted P. PD184352 (CI-1040) ananatis pqq operon led to the inability of mutant cells to accumulate gluconic (Table 4) acid and to the abolition of GDH activity in their extracts (Table 2) without exogenous PQQ in reaction in distinction from GDH extracted from P. ananatis SC17(0). Thus, it was confirmed that PQQ is indeed essential for the formation of active holoenzyme GDH and predicted pqq operon encoded genetic elements essential for PQQ biosynthesis. Construction of the strain SC17(0)-φ80attB-pqq was achieved by in vivo cloning of the pqq operon (see Supporting Information) and adaptation of ‘Dual In/Out’ Recombineering-driven strategy for the integration of DNA fragments into targeted points of the E. coli chromosome (Minaeva et al., 2008) for application in P. ananatis. The scheme applied in this work could be a useful instrument for the simple amplification of target genes/operons in the chromosome without preliminary amplification by PCR. The resulting strains, SC17(0)-Δpqq with the ΔpqqABCDEF operon and SC17(0)-φ80attB-pqq with two copies of this operon in the chromosome, were tested for their ability to accumulate PQQ in cultural medium. Inactivation of the putative pqq operon resulted in a decrease of PQQ in the medium to undetectable levels (<1 mg L−1).

While this is an important work, it does not fully explain the slow and incomplete transition towards patient-centred care. We wonder if pharmacists’ own mental barriers are a missing piece. In our comparison of two legislatively progressive jurisdictions, community pharmacists in Northern Ireland provided more patient-centred responses find more than community pharmacists in Alberta (P = 0.013), although both described product-focused roles in 39–45% of their responses. The product focus of pharmacists was also borne out in the word-cloud analyses, with very little use of patient-care terminology to describe what a pharmacist does. To our knowledge this is the first study to use short telephone

interviews which elicit a ‘top of mind’ or automatic response to compare how community pharmacists from Alberta and Northern Ireland describe what a pharmacist does. This approach engages certain

unconscious mental processes which affect and influence the judgements, feelings and behaviours of the person.[35] In the literature it has been reported that individuals’ automatic response does not usually match their self-reported attitudes.[36] The slight deception and restriction of response were intended to remove some of the effects of social desirability bias.[37] We think that our findings are generalisable to pharmacy practice in Alberta and Northern Ireland because the key demographic features of our samples are similar to regional averages (Table 3). A potential limitation of the present study relates to the fact that pharmacists’ responses were restricted LEE011 by the study question and our request for a brief response. If they had more time to think about their responses there is a chance that they would have been different. Nevertheless, the intention of using this methodology was to prevent pharmacists from thinking too much about their answer, thereby eliciting a ‘top of mind’ or automatic response and to avoid some of the effects of social desirability bias. Another potential limitation is the use of word clouding which represents a visualisation of Forskolin solubility dmso the frequency

of the reported words. This method may not take into account the context in which the words were used. Also the use of open questions has the potential to introduce recall bias as this approach assumes that if a term was not reported then that term is not relevant. The higher degree of patient-centred responses provided by Northern Ireland pharmacists might be explained by the differences in contracts and payment schemes between Northern Ireland and Alberta. In Northern Ireland community pharmacists are paid for offering certain patient-centred services such as smoking cessation and minor ailments management,[33] while in Alberta (and Canada in general) the current model of reimbursement provides pharmacists with dispensing fees only (as in the traditional system of practice).

The effects http://www.selleckchem.com/products/Roscovitine.html of a change of location were investigated for the day

prior to CoR (CoR−1), the CoR (CoR0, eg, travel day), and the first day at the new location (CoR+1). The fifth day after the change of residence (CoR+5) was used as a post-CoR reference value. Perceived travel strain was measured with a 4-point worded scale [“travel strain was very (4), rather (3), hardly (2), not at all (1) strenuous”]. To test for the adequacy of the given sample size, a statistical power calculation was conducted using the power calculator provided by our University, imputing the baseline and average response values. The statistical power of the three significant variables was 0.26/0.36/0.90 (systolic BP/diastolic BP/sleep), indicating a small power for detecting differences in BP, but a large power for detecting differences in sleep. To test for the feasibility of using a parametrical statistical approach, the normal distribution of all four dependent variables (diastolic BP, systolic BP, quality of sleep, and mood) during pre-travel baseline and on the four single days around the CoR was controlled for visually on the basis of histograms. All distributions were found to be adequate. To analyze the effect

of the CoR, a multivariate analysis of variance for repeated measures was Navitoclax calculated for the five time points BL, CoR−1, CoR0, CoR+1, and CoR+5, thereby comparing each of the days CoR−1 to CoR+5 with the baseline value HA-1077 clinical trial (BL) using so-called “simple contrasts.” Thus, four contrasts were calculated for every variable. The statistical significance of these comparisons (p values) is displayed in Table 2. All four outcome variables were analyzed simultaneously in the multivariate approach, thus following the suggestions of Drummond to use one global statistical test.[38] Also, this approach controlled for the multiple comparisons calculated. To test for possible differences between morning and evening

BP readings, average morning and evening BP responses (average of CoR−1, CoR0, CoR+1 − BL) were compared using t-tests for paired samples. To test the association of the responses to the CoR with variables describing the study participants, their medical condition and travel, the correlation of the response values (average of CoR−1, CoR0, CoR+1 − BL) of BP, sleep, and mood with these variables was calculated. Also, the inter-correlation of the average responses of the four outcome variables to the CoR was determined. To test the validity of the scales used, their correlation with standardized scales, clinical BP readings, or other external variables was calculated. All analyses were conducted using SPSS 15.0. The results illustrated in Figure 1 are based on means and confidence intervals.

The effects Compound Library concentration of a change of location were investigated for the day

prior to CoR (CoR−1), the CoR (CoR0, eg, travel day), and the first day at the new location (CoR+1). The fifth day after the change of residence (CoR+5) was used as a post-CoR reference value. Perceived travel strain was measured with a 4-point worded scale [“travel strain was very (4), rather (3), hardly (2), not at all (1) strenuous”]. To test for the adequacy of the given sample size, a statistical power calculation was conducted using the power calculator provided by our University, imputing the baseline and average response values. The statistical power of the three significant variables was 0.26/0.36/0.90 (systolic BP/diastolic BP/sleep), indicating a small power for detecting differences in BP, but a large power for detecting differences in sleep. To test for the feasibility of using a parametrical statistical approach, the normal distribution of all four dependent variables (diastolic BP, systolic BP, quality of sleep, and mood) during pre-travel baseline and on the four single days around the CoR was controlled for visually on the basis of histograms. All distributions were found to be adequate. To analyze the effect

of the CoR, a multivariate analysis of variance for repeated measures was LEE011 research buy calculated for the five time points BL, CoR−1, CoR0, CoR+1, and CoR+5, thereby comparing each of the days CoR−1 to CoR+5 with the baseline value Decitabine (BL) using so-called “simple contrasts.” Thus, four contrasts were calculated for every variable. The statistical significance of these comparisons (p values) is displayed in Table 2. All four outcome variables were analyzed simultaneously in the multivariate approach, thus following the suggestions of Drummond to use one global statistical test.[38] Also, this approach controlled for the multiple comparisons calculated. To test for possible differences between morning and evening

BP readings, average morning and evening BP responses (average of CoR−1, CoR0, CoR+1 − BL) were compared using t-tests for paired samples. To test the association of the responses to the CoR with variables describing the study participants, their medical condition and travel, the correlation of the response values (average of CoR−1, CoR0, CoR+1 − BL) of BP, sleep, and mood with these variables was calculated. Also, the inter-correlation of the average responses of the four outcome variables to the CoR was determined. To test the validity of the scales used, their correlation with standardized scales, clinical BP readings, or other external variables was calculated. All analyses were conducted using SPSS 15.0. The results illustrated in Figure 1 are based on means and confidence intervals.

Int this website J Cancer 2003; 103: 142–144. 18 Mocroft A, Kirk O, Clumeck N et al. The changing pattern of Kaposi sarcoma in patients with HIV, 1994–2003: the EuroSIDA Study. Cancer 2004; 100: 2644–2654. 19 Engels EA, Pfeiffer RM, Goedert JJ et al. Trends in cancer risk among people with AIDS in the United States 1980–2002. AIDS

2006; 20: 1645–1654. 20 Franceschi S, Maso LD, Rickenbach M et al. Kaposi sarcoma incidence in the Swiss HIV Cohort Study before and after highly active antiretroviral therapy. Br J Cancer 2008; 99: 800–804. 21 Guiguet M, Boué F, Cadranel J et al. Effect of immunodeficiency, HIV viral load, and antiretroviral therapy on the risk of individual malignancies (FHDH-ANRS CO4): a prospective cohort study. Lancet Oncol 2009; 10: 1152–1159. 22 Selik RM, Byers RH Jr, Dworkin MS. Trends in diseases reported on U.S. death certificates that mentioned HIV infection, 1987–1999. J Acquir Immune Defic Syndr 2002; 29: 378–387. 23 Simard EP, Pfeiffer RM, Engels EA. Cumulative incidence of cancer among individuals with acquired immunodeficiency syndrome in the United States. Cancer 2011; 117: 1089–1096. 24 Lodi S, Guiguet M, Costagliola D et al. Kaposi sarcoma incidence Caspase activity assay and survival among HIV-infected homosexual men

after HIV seroconversion. J Natl Cancer Inst 2010; 102: 784–792. 25 Pipkin S, Scheer S, Okeigwe I et al. The effect of HAART and calendar period on Kaposi’s sarcoma and non-Hodgkin lymphoma: results of a match between an AIDS and cancer registry. AIDS 2011; 25: 463–471. 26 Shiels MS, Pfeiffer RM, Gail MH et al. Cancer burden in the HIV-infected population in the United States. J Natl Cancer Inst 2011; 103: 753–762. 27 Sitas F, Carrara H, Beral V et al. Antibodies

against human herpesvirus 8 in black South African patients with cancer. N Engl J Med 1999; 340: 1863–1871. 28 Bassett MT, Chokunonga E, Mauchaza B et al. Cancer in the African population of Harare, Zimbabwe, 1990–1992. Int J Cancer 1995; 63: 29–36. 29 Wabinga HR, Parkin DM, Wabwire-Mangen F, Nambooze S. Trends in cancer incidence in Kyadondo County, Uganda, 1960–1997. Br J Cancer 2000; 82: 1585–1592. 30 Parkin DM, Sitas F, Chirenje M et al. Part I: Cancer DOCK10 in indigenous Africans–burden, distribution, and trends. Lancet Oncol 2008; 9: 683–692. 31 Mosam A, Carrara H, Shaik F et al. Increasing incidence of Kaposi’s sarcoma in black South Africans in KwaZulu-Natal, South Africa (1983–2006). Int J STD AIDS 2009; 20: 553–556. 32 Chokunonga E, Borok MZ, Chirenje ZM et al. Trends in the incidence of cancer in the black population of Harare, Zimbabwe 1991–2010. Int J Cancer 2013; 133: 721–729. 33 Mosam A, Uldrick TS, Shaik F et al. An evaluation of the early effects of a combination antiretroviral therapy programme on the management of AIDS-associated Kaposi’s sarcoma in KwaZulu-Natal, South Africa. Int J STD AIDS 2011; 22: 671–673. 34 Casper C.