, 1999). An in vivo study showed that 6 weeks after the administration of a garlic extract, H. pylori-induced Selleckchem Autophagy inhibitor gastritis in Mongolian gerbils was decreased in a dose-dependent manner compared with the control group, even though the number of viable H. pylori was not changed (Iimuro et al., 2002). Several epidemiological studies suggested that a decreased risk of gastric cancer is associated with an increasing consumption of allium vegetables (You et al., 1989), possibly due to an effect on H. pylori, as this organism is associated

with gastric cancer. Thus, it is very important to study the antibacterial mode of action of garlic constituents because of the high incidences of H. pylori-related diseases throughout the world. Although previous studies have revealed that the antimicrobial effect of garlic is mainly due to its chemical reaction with thiol groups of various enzymes, such as alcohol dehydrogenase

and thioredoxin reductase (Ankri & Mirelman, 1999), a detailed analysis is lacking of the global molecular responses induced by garlic and its derivatives, and a proteomic strategy is required to globally profile the cellular MS-275 ic50 responses at the protein level under defined conditions. Allitridi, whose chemical constituent is diallyl trisulfide (DATS), is a proprietary garlic derivative and has been successfully used to treat both systemic fungal and bacterial infections in China (Shen et al., 1996). In the present investigation, to obtain a comprehensive picture of the antibacterial mode of action of allitridi in H. pylori, proteomic analysis was used to study the global protein alterations induced by allitridi treatment. In addition, the effects of allitridi on virulence factor production by H. pylori were also investigated at subinhibitory concentrations. Helicobacter pylori

26695 was kindly provided by Dr Zhang Jianzhong from the Chinese Disease Control and Prevention Center. Allitridi was obtained from Jinan Limin Pharmaceutical Co., Ltd (Shandong, China). The bacteria were cultured to the early exponential phase in Brucella broth containing 10% fetal bovine serum with 120 r.p.m. shaking in a microaerobic environment (5% O2, 10% CO2 and 85% N2) at 37 °C. Aliquots of 10 mL of the cell cultures were supplemented with a series of concentrations of allitridi Metformin mouse to examine its inhibitory effect. To assay viability at each time point (0, 6, 12 and 24 h), the number of CFU was determined by plating serial dilutions of cultures in duplicate on Skirrow agar plates with 5% (v/v) sheep blood. Each assay was replicated at least three times. Exponentially growing H. pylori supplemented with or without 1 μg mL−1 allitridi for 6 h were harvested by centrifugation and resuspended in lysis buffer containing 8 M urea, 2 M thiourea, 4% CHAPS, 1% dithiothreitol, 1% pharmalyte (pH range 3–10), 1% protease inhibitor and 1% nuclease mix (Amersham Biosciences).

In the UK, the virological failure rate on current first-line regimens in 2008–2009 was approximately 10% at 1 year [2]. The options for switch depend on the most recent and past ARV treatments as well as current and archived resistance results. As genotypic testing in ARV-naïve patients is now performed routinely and is recommended practice, detection of resistance at virological failure is rarely a result of transmitted drug resistance and failure to

adapt first-line treatment [3, 4]. The general principles for the management of patients experiencing virological failure are outlined in Boxes 1 and 2 as GPPs. Details of typical patterns of HIV drug resistance found in patients with a history of or presenting with virological failure are outlined in Box 3. For guidance on HIV VL, drug selleck compound resistance and tropism testing, the reader should consult the BHIVA routine investigation and monitoring guidelines [1]. Factors affecting adherence and drug exposure, including tolerability/toxicity issues, DDIs/food interactions, ARV potency, significant renal/liver disease and mental health/drug dependency problems are evaluated. Resistance testing is performed while on failing therapy or within 4 weeks of discontinuation. Past ART and resistance tests are reviewed for archived mutations. Tropism testing is performed if MVC is being considered. Intensification with an additional

active ARV is not recommended. Once virological failure is confirmed and a resistance result available, this website the regimen is changed as soon as possible to avoid accumulation of resistance mutations. The choice of the new ART regimen will primarily depend on the results of resistance testing and the patient’s preference. Montelukast Sodium Additional considerations include the results of tropism and HLA-B*57 testing, DDIs/food interactions, co-morbidities and future therapy options. The goal of the new combination is to re-establish a VL <50 copies/mL. In patients with ongoing viraemia and with few options to construct

a fully suppressive regimen, referral for specialist advice and/or discussion in a multidisciplinary team ‘virtual’ clinic. Include at least two and preferably three fully active agents with at least one active PI/r (e.g. DRV/r) and one agent with a novel mechanism of action (CCR5 antagonist/integrase or fusion inhibitor). Treatment interruption is not recommended. No resistance (WT virus). 3TC/FTC resistance (M184V/I) following any first-line therapy, including TDF/FTC or ABC/3TC. NNRTI resistance (e.g. K103N or Y181C/I/V) and/or 3TC/FTC resistance (following first-line therapy with NNRTI-based regimen, including TDF/FTC or ABC/3TC). INI resistance (e.g. Q148 or N155H) and/or 3TC/FTC resistance (following first-line therapy with RAL-based regimen, including TDF/FTC or ABC/3TC). Extended RT resistance (e.g. K65R/L74V or thymidine) (following suboptimal regimens/patients with more extensive drug history associated with virological failure).

“
“The polysaccharide capsule of Streptococcus pneumoniae is the main virulence GSK126 factor making the bacterium resistant to phagocytosis. The galU gene of S. pneumoniae encodes a UDP-glucose pyrophosphorylase absolutely required

for capsule biosynthesis. In silico analyses indicated that the galU gene is co-transcribed with the gpdA gene, and four putative promoter regions located upstream of gpdA were predicted. One of them behaved as a functional promoter in a promoter reporter system. It is conceivable that the sequence responsible for initiating transcription of gpdA-galU operon is an extended −10 site TATGATA(T/G)AAT. Semi-quantitative real-time reverse transcription PCR experiments indicated that galU was expressed mainly in the exponential phase of growth. Streptococcus pneumoniae is a leading human pathogen causing both mucosal (such as otitis media and pneumonia) and systemic diseases (including septicemia and meningitis). To date, 93 different pneumococcal see more capsular types have been described (Henrichsen, 1995; Park et al.,

2007; Bratcher et al., 2010; Calix & Nahm, 2010). This remarkable phenotypic variability appears to be present also at the genetic level (Bentley et al., 2006). Early studies showed that uridine diphosphoglucose (UDP-Glc) is a key component in the Liothyronine Sodium biosynthetic pathway of pneumococcal capsular polysaccharides containing glucose, galactose, and/or UDP-glucuronic or UDP-galacturonic acids (Mills & Smith, 1965).

At least one of these sugars is a component of every capsular polysaccharide of S. pneumoniae (Kamerling, 2000). The enzyme UTP-Glc-1-phosphate uridylyltransferase (UDP-Glc pyrophosphorylase; EC 2.7.7.9) is encoded by the galU gene. This enzyme catalyzes the formation of UDP-Glc, which is the substrate for the synthesis of UDP-glucuronic acid. Also, UDP-Glc is also required for the interconversion of galactose and glucose by way of the Leloir pathway (Frey, 1996). Previously, the galU gene was cloned and overexpressed, and the gene product was biochemically characterized (Mollerach et al., 1998; Bonofiglio et al., 2005). In addition, knockout galU mutants of type 1 and type 3 pneumococci are unable to synthesize a detectable capsular polysaccharide. Southern blot hybridization experiments using DNAs prepared from pneumococcal isolates belonging to different types showed that every strain tested contained a galU homologue (Mollerach et al., 1998). Thus, the UDP-Glc pyrophosphorylase, which is directly involved in the synthesis of the capsular polysaccharide in S. pneumoniae, might represent a suitable target in the search for inhibitors to control the biosynthesis of the main pneumococcal virulence factor.

These programmes can Dabrafenib chemical structure range from 540 to 2145 contact hours (24–87 weeks), with a median of 940 h. The ASHP requires that programmes have minimum of 600 contact hours and a minimum duration of 15 weeks to apply for accreditation, and, as of January 2009, 147 technician

training programmes have sought such accreditation.[17] These accredited programmes will have trained an estimated 12 000 technicians in 2009, with greater than half of all graduates representing the three largest retail drug stores in the country. It should be noted that there are also numerous unaccredited online programmes that exist for the training of pharmacy technicians, but which lack any uniform educational or practical training components.[17] The Model Curriculum for Pharmacy Technician Training, a curriculum developed by the ASHP in conjunction with the APhA, the American Association of Pharmacy Technicians, the Pharmacy Technician Educators Council, the American Association of Colleges of Pharmacy and the National Cyclopamine concentration Association of Chain Drug Stores has been a positive step towards a standardized model of training.[22] The first edition, based on a task analysis performed by the PTCB, was created in 1997 and updated in 2001.[35] There

have been significant changes in areas dealing with the technician’s role in safe medication use, assisting with immunizations and the institutional use of the ‘tech-check-tech’ system, in which pharmacy technicians, rather than the pharmacist, are responsible for validating the Silibinin work of other technicians and serve as the final check in the filling process.[30] The goal for the curriculum is to provide a menu of possible learning outcomes and it provides suggestions for competencies that one would expect a pharmacy technician to be well-versed in (e.g. anatomy and physiology, basic therapeutics, pharmacology). It does not make any recommendations regarding length of training.[36] Development of standardized education would not eliminate the need for on-the-job training or education

pertaining to local policies and procedures.[10] Two types of accreditation currently exist. The first is programmatic, or specialized, accreditation, which focuses on individual programmes. Initially, nearly all accredited programmes were hospital-based.[20] Currently only three technician training programmes are hospital-based, and 90% of programmes are located at vocational, technical or community colleges.[10] The second type of accreditation, institutional, evaluates the institution as a whole. Four agencies carry out this type of accreditation. None have a formal national affiliation with the profession of pharmacy.[10] Completion of an accredited programme is not usually a requirement for employment, registration or certification of pharmacy technicians.

The ligated RNAs were size-fractionated on a 15% TBE–urea polyacrylamide gel and the RNA fragments of c. 41–76 nt in length were isolated. The SRA 3′-adapter (Illumina) ligation was then performed followed by a second size-fractionation using TSA HDAC manufacturer the same gel conditions as described above. The

RNA fragments of c. 64–99 nt in length were isolated through gel elution and ethanol precipitation. The ligated RNA fragments were reverse transcribed to single-stranded cDNAs using M-MLV reverse transcriptase (Invitrogen, Carlsbad, CA) with RT primers recommended by Illumina. The cDNAs were amplified with pfx DNA polymerase (Invitrogen) in 20 cycles of PCR using Illumina’s sRNA primers set. PCR products were run on a 12% TBE polyacrylamide gel and a slice of gel containing fragments of c. 80–115 bp in length was excised. This fraction

was eluted and the recovered cDNAs were precipitated and quantified using a Nanodrop (Thermo Scientific, Rockford, IL) and a TBS-380 mini-fluorometer (Turner Biosystems, Sunnyvale, CA) using Picogreen dsDNA quantization reagent (Invitrogen). The sample concentration was adjusted to c. 10 nM and a total of 10 μL was used in the sequencing reaction. The purified cDNA library was used for cluster generation on Illumina’s Cluster Station and then sequenced on Illumina Genome Analyzer IIx following the supplier’s instructions for running the instrument. Raw sequences were processed using Illumina’s Pipeline software and selleckchem then subjected to a series of data filtration steps to analyse sequencing data using the ACGT101-miR software package (V3.5; LC Sciences). The reference database of S. mutans UA159 (http://www.ncbi.nlm.nih.gov/nuccore/AE014133) and Rfam (http://rfam.sanger.ac.uk) was used for msRNA mapping. Hairpin RNA structures were predicted from the adjacent 60–80 nt sequences in either

direction using mfold software (Zuker, 2003). Real-time quantitative RT-PCR (qRT-PCR) was performed to verify the presence of several selected candidates within the fraction of purified cellular RNAs. The total RNA (50 ng) was reverse transcribed using a TaqMan microRNA Reverse Transcription PAK5 kit. From the 15 μL of RT mixture, 2 μL was used for real-time PCR. qRT-PCR was performed with TaqMan universal master mix (Applied Biosystems, Foster City, CA). Seventeen msRNAs were selected and specific primer sets and TaqMan probes were designed by Applied Biosystems. Ten out of 17 custom-designed TaqMan probes and primer sets failed, which may be due to the small size or structure of verified RNA species. PCR was carried out in 96-well plates using the 7500 Real-Time PCR system (Applied Biosystems). The expression of each msRNA gene was determined from three replicates in a single qRT-PCR experiment. The total RNA (20 μg) was separated on a 15% urea-acrylamide gel and blotted onto nylon N+ membrane (Invitrogen).

, 2000). A higher number of replicates could have allowed the isolation of numerous other bacterial taxa as suggested by DGGE that explored the biodiversity of bacterial communities present on AMF spores collected Selleckchem NU7441 from a field. Moreover, Scheublin et al. (2010) reported that bacteria from the Oxalobacteraceae family are abundant and adhere to AMF hyphae. Other reports hypothesized that Oxalobacteraceae may specifically interact with mycorrhizal fungi (Pivato et al., 2009). Lioussanne et al. (2010) have found different Pseudomonas

spp., Herbaspirilium sp., Acidobacterium sp., Bacillus spp. and Verrucomicrobium sp. specifically associated with G. irregulare or Glomus mosseae. However, although we isolated this website three members of the Bacillus genus in our study, they were less abundant and represent only six morphotypes in contrast to V. paradoxus, which represents 13 morphotypes. Using DGGE, we assessed the bacterial biodiversity of washed spores of G. irregulare isolated from soil (Fig. 4). DGGE patterns from three field-collected spores were markedly different in the number

of bands formed, but mainly in their migration positions, indicating a widely different community structure between spores. The number of bands ranged from 17 to 24, and although 29–41% of the band positions were common to all spores, 28–38% were unique to each one. The markedly variable banding pattern seen on the DGGE clearly shows that a much higher number of bacterial taxa were associated with the spores than the number suggested by

isolation. Soils may contain noncultivable taxa or taxa with specific nutritional needs that were not met with the isolation protocol used in this study. Some of the bacterial Progesterone isolates recovered from the spores were also analyzed in DGGE. However, our DGGE analyses confirmed that we isolated only a very small proportion of bacterial taxa living on the surface of AMF spores. In addition, bacterial community structure varies considerably among spores. It should be remembered that we cannot be certain whether these spores originate from the same location because we mixed six samples taken from six sites spread along a 50-m distance to reduce the bias due to variation in the local composition of the soil. Soil biotic and abiotic conditions where each AM spore used was taken could then differ markedly and change the bacterial pattern associated with AMF spore. When we monitored the interactions between the isolated bacteria and the mycelium of G. irregulare based on morphological growth and adherence, we found that all bacterial taxa except for B. cereus grew and adhered on the surface of hyphae and spores (Table S1 and Fig. 5). The growth rates and patterns were, however, different between taxa. For example, B. cereus was rarely detected and grew very slowly. In contrast, B.

02). In the HIV-positive group, prior or current AIDS-defining events were reported for 30% of patients, 9% and 30% had CD4 counts of <200 and 200–500 cells/μL, respectively,

and 95% had HIV-1 RNA <50 copies/mL. Pneumonia (9%vs. 25%, respectively, in the HIV-positive and HIV-negative groups; P=0.01) and respiratory failure (9%vs. 21%, respectively; P=0.04) were less common in the HIV-positive group. Oseltamivir BIBW2992 datasheet (95%vs. 71% in the HIV-positive and HIV-negative groups, respectively; P=0.003) was administered more often in HIV-positive patients. Three patients (all HIV-negative) died. In the HIV-positive group, CD4 cell count and plasma HIV-1 RNA did Selisistat not differ before and 4–6 weeks after influenza A H1N1 diagnosis (P>0.05). HIV infection did not increase the severity of influenza A H1N1 infection, and influenza A H1N1 infection did not have a major effect on HIV infection.

Influenza is a common cause of acute respiratory illness in HIV-infected adults [1,2]. Before the widespread use of effective combination therapy, small case series and anecdotal reports suggested that low CD4 cell counts or concomitant respiratory or cardiovascular comorbidities were associated with a higher risk for complications [3–8]. It is unclear to what extent effective antiretroviral therapy may have affected the risk for severe or complicated influenza, but HIV-infected patients are still considered to be Tyrosine-protein kinase BLK at a higher risk [9] and for that reason they are preferentially targeted for influenza vaccination [10–12]. Human infections with a novel A H1N1 influenza virus were first identified in April 2009 [13,14] and they were increasingly reported throughout the world in the following weeks [15]. The rapid spread of the infection and the extensive reporting of associated deaths occupied the attention of the media and contributed to increased awareness

in the general population [16,17]. Data from the beginning of the epidemics suggested that many influenza A H1N1 infections were not necessarily severe [18] and that HIV-infected patients were not overrepresented among those hospitalized or severely ill [14,19–21]. Nevertheless, health authorities considered that HIV-infected patients were at a higher risk for influenza A H1N1 complications, as they were for seasonal influenza, and this assumption remains unchanged [22–24]. With open access to combination antiretroviral therapy, many HIV-infected adults show sustained suppression of HIV replication in plasma, resulting in immunological and clinical benefits [25]. In Spain, health care for chronic conditions such as HIV infection and also for acute conditions and emergencies is provided free of charge by the public health care system [26].

Conidia are ellipsoidal to ovoid or subcylindrical, thin and smooth-walled, hyaline, aseptate to septate, extremely variable in size [(5) 5.5–9.5 (10) μm (x=7.05, SD=1.18, n=30) × (3) 3.5–4.5 (5) μm (x=4.26, SD=0.64, n=30)] and rarely guttulate (Fig. 1). Collectively, these morphological features strongly support the placement of the present isolate as a species of Phoma Sacc. emend. Boerema & G.J. Bollen (Fig. 1). Furthermore,

ITS sequence data showed that the endophyte is a strain of the genus Phoma (Fig. 2). The ITS 5.8S ribosomal Navitoclax gene showed a maximum homology of 99.2% with Phoma herbarum strain BLE15 and Phoma sp. strain 11360. The endophyte also exhibited 99% sequence homology with Phoma medicaginis strain CBS 533, Phoma macrostoma, Ascochyta rabiei (Phoma rabiei) strain CBS 237.37 and Didymella phacae CBS strain 184.55, as presented in the distance matrix chart (Fig. 2). No Phoma sp. previously has been reported

from this plant either as an endophyte or as a pathogen. The genus Phoma sp., as typified by P. herbarum (Boerema 1964), is a complex and heterogeneous assemblage of more than 3000 infrageneric taxa (Monte et al., 1991). It has been considered to be one of the largest fungal genera, consisting of taxa inhabiting soil, organic debris and water, as well as species that parasitize Ivacaftor datasheet other fungi, lichens, insects and vertebrates. In addition, a substantial proportion of the taxa are associated with plant material as primary pathogens. In the case of isolate Ut-1, it appears that the fungus can exist in the host plant as both an endophyte and a pathogen under some circumstances. It was possible to show pathogenicity of

the organism on inoculated leaves of the host, yielding necrotic spots. Also, subsequently it was possible to successfully reisolate the causal agent using standard procedures followed by identification of the organism on the basis of its morphological features (Fig. 1). When Phoma sp. was grown on PDA for 10–12 days and the headspace was examined for VOC content the most significant observation was that at least 15 compounds appeared whose mass was 204 and Avelestat (AZD9668) whose chemical assignment was that of a sesquiterpene, with α-humulene (or α-caryophyllene) being the most predominant VOC (Table 1). Furthermore, trans-caryophyllene is also present in the fungal VOC headspace and it too is a major VOC in the volatiles of L. tridentata (G. Strobel, unpublished data). Also of interest is the presence of a number of reduced naphthalene derivatives such as those with retention times of 15.06, 15.12, 16.31 and 18.68 min (Table 1). Reduced naphthalene compounds of this type have been reported from M. albus (Strobel et al., 2001). GC/MS analyses of diesel fuel from all parts of the world have revealed the presence of reduced and sometimes derivatized naphthalenes of the general type produced by Phoma sp. (Adams & Richmond, 1951; G. Strobel, unpublished data).

Overall, 180 additional NNRTI mutations were found to have accumulated over 295 years [1 new/1.6 years; 95% confidence interval (CI) 1.5–1.8]. The rate of accumulation was faster ICG-001 cell line in the first 6 months from VF (1 new/1.1 years), and slower in patients exposed to nevirapine vs. those receiving efavirenz [relative risk (RR) 0.66; 95% CI 0.46–0.95; P=0.03]. There is an initial phase of rapid accumulation of NNRTI mutations close to the time of VF followed by a phase of slower accumulation. We predict that it should take approximately one year of exposure to a virologically failing first-generation NNRTI-based cART regimen to reduce

etravirine activity from fully susceptible to intermediate resistant, and possibly longer in patients kept on a failing nevirapine-containing regimen. Global access to antiretroviral drugs has increased dramatically in recent years [1], and concerns regarding the development of drug resistance remain in both resource-rich and resource-limited settings [2,3]. In resource-limited settings, NNRTIs are a fixed component of first-line combination antiretroviral therapy (cART) [3], but HIV-infected populations typically have little access to virological

monitoring and/or genotypic resistance testing, which is likely to result in the accumulation of NNRTI resistance. An improved access to NNRTI drugs for preventing APO866 price mother-to-child transmission has further complicated this issue. A previous analysis of patients in EuroSIDA focused on the estimation of the rate of accumulation of thymidine analogue mutations (TAMs) in patients kept on zidovudine or stavudine despite

Cytidine deaminase a viral load of >500 HIV-1 RNA copies/mL [4,5]. NNRTI resistance accumulation could compromise the efficacy of second-generation NNRTIs (e.g. etravirine [6]) if they ever become available in these settings. Indeed, etravirine has already been used in some resource-limited settings as a component of second-line regimens in patients who could not tolerate protease inhibitors (PIs) [7]. Data on etravirine resistance in patients already exposed to first-generation NNRTIs show that, among 17 mutations in the reverse transcriptase gene, at least three must be present simultaneously in order to reduce etravirine activity, although just two mutations can greatly decrease susceptibility in some cases [7–9]. In addition, this activity is likely to diminish to zero as NNRTI-associated resistance mutations further accumulate. Our analysis is based on data for patients enrolled in clinics in Europe. However, while there are differences in the prevalence of HIV subtypes, some infections and in access to health care between resource-rich and resource-limited settings, there is otherwise generally little evidence of differences between these settings in the damage caused by HIV or the effect of ART [10–12].

Pyrimethamine is a folate antagonist and should be prescribed with folinic acid. Alternative options are clindamycin (B) with pyrimethamine (C) or atovaquone (C). Secondary prophylaxis should be as for the non-pregnant. All pregnant women should have T. gondii serological status checked. In the non-immunocompromised host, transmission of T. gondii to the foetus usually only occurs during acute infection. However, there have been case

reports of transmission following reactivation in HIV-infected women with severe immunosuppression [21], although this is rare. Where there is evidence of acute infection or symptomatic reactivation in the mother, the foetus should be screened for evidence of perinatal transmission. Studies following up immunocompetent women with acute toxoplasmosis www.selleckchem.com/products/sch772984.html in pregnancy have not shown any conclusive evidence

for the effectiveness of spiramycin, or sulphadiazine with pyrimethamine, to prevent congenital foetal infection [41,42]. For systemic disease systemic therapy will be required. However, for patients with single site retinal disease, consideration may be given to providing local intravitreal therapy or implants to reduce foetal exposure to antivirals. All the available antiviral agents, ganciclovir (C), valganciclovir (C), foscarnet (C) and cidofovir (C), are associated with congenital anomalies in rats and rabbits [43,44]. Ganciclovir is embryotoxic learn more in rabbits and mice and teratogenic in rabbits. There is no published experience of valganciclovir

in pregnancy, but the same concerns exist as for ganciclovir. Foscarnet is associated with an increased risk of skeletal anomalies in rats and rabbits, but there is no experience of its use in early human pregnancy. Due to the potential for renal toxicity, careful monitoring of amniotic fluid should be undertaken, very especially in the second and third trimester, for oligohydramnios. Cidofovir also has shown evidence of embryotoxicity and teratogenicity in rats and rabbits, and there is no experience of using this drug in pregnancy. Therefore, the most experience in clinical practice has been with intravenous ganciclovir, and either this agent or oral valganciclovir should be considered first line treatment for CMV disease in pregnancy [45,46]. Infants born to mothers with evidence of active CMV disease should be examined for evidence of congenital infection [18]. Oral aciclovir (B) for either acute attacks or prophylaxis is indicated [47]. No adverse outcomes have been reported to the infant after in utero exposure to this drug [48,49]. There are fewer registry data available for famciclovir (B) or valaciclovir (B), and the manufacturers recommend their use only when potential benefits outweigh the risk [50]. HIV infection and tuberculosis are closely linked; HIV infection increases the risk of reactivation of latent TB by at least 20 fold [51,52].