Injured travelers as well as medical tourists are directly concer

Injured travelers as well as medical tourists are directly concerned by this strategy. This article has been kindly proofread by Amy Whereat, Medical English Consultant. The authors state they have no conflicts of interest to declare. “
“A 34-year-old Nigerian man presented with nephrotic syndrome. Renal biopsy revealed chronic membranous glomerulopathy with focal segmental sclerosis. Blood Giemsa smear contained rare Plasmodium sp. trophozoites and small subunit

ribosomal RNA polymerase chain reaction amplification confirmed the presence of Plasmodium malariae. This case highlights the importance of obtaining even remote travel histories from ill immigrants and considering occult quartan malaria in patients from endemic locations with nephrotic syndrome. Although quartan ABT-737 nmr malaria comprises only a small portion of the global disease burden from malaria, Plasmodium this website malariae is unique among the plasmodia in which subclinical parasitemia may persist for decades with illness occurring more than 40 years after the last possible exposure.1 Additionally, chronic P malariae infection was linked to nephrotic syndrome in children in the 1960s and subsequently attributed to immune complex basement membrane nephropathy.2,3 We describe a case of P malariae-associated chronic membranous glomerulopathy and nephrotic

syndrome in a US Navy sailor 14 years after his last possible exposure to the risk of malaria. This case highlights the importance of obtaining remote travel histories from Phospholipase D1 immigrants presenting with illness, even decades after emigration from their country of origin. A 34-year-old US-born African American Navy sailor, who moved to Nigeria at the age of 1, migrated back to the United

States at the age of 21 and had not traveled home or to any malaria endemic locations during the ensuing 14 years. While at sea, he presented to his ship’s medical doctor with a 4-month history of bilateral lower extremity pitting edema and swelling of his face and a 5-month history of frothy urine. He was notably hypertensive with hyperlipidemia (total cholesterol 390 mg/dL, low density lipoprotein 305 mg/dL, triglycerides 230 mg/dL) and was placed on hydrochlorothiazide and simvastatin. Upon return to port, the patient was referred to Internal Medicine for suspected nephrotic syndrome. His past medical history was significant for sickle trait, treated latent tuberculosis, and childhood malaria. He denied a family or personal history of kidney disease. Laboratory studies were significant for a spot protein/creatinine ratio of 22.6, consistent with nephrotic syndrome. Additional abnormal laboratory findings included low serum albumin (1.8 g/dL), high serum creatinine (6.2 mg/dL), and a low glomerular filtration rate (14 mL/min).

This indicates that a classifier trained only on pictures of sepa

This indicates that a classifier trained only on pictures of separately presented faces and places may not be the most optimal way of decoding object-based visual attention. Concluding, we have shown that real-time fMRI allows for online prediction of attention to objects belonging to different object categories. Prediction is based on distributed patterns of activity in multiple brain regions. The outlined methodology not only allows us to probe object-based attention in an online setting Selleckchem OSI 906 but also illustrates the potential to develop BCIs that are driven

by modulations of high-level cognitive states. The authors gratefully acknowledge the support of the BrainGain Smart Mix Programme of the Netherlands Ministry of Economic Affairs and the Netherlands Ministry of Education, Culture and Science. The first

author was supported by a UTS grant from the University of Twente. We thank Paul Gaalman for his technical support during the experimental setup and development of the real-time fMRI pipeline. We are very grateful to the editors and the anonymous reviewers for their encouraging and constructive comments on our manuscript. Abbreviations aMTG anterior medial temporal gyrus BCI brain–computer interface BOLD blood oxygen level-dependent FFA fusiform face area fMRI functional magnetic resonance imaging GLM general linear model MoCo motion-corrected MVA-C cluster-wise learn more multivariate analysis MVA-G GLM-restricted multivariate analysis MVA-T mean timeseries multivariate analysis MVA-W whole-brain multivariate analysis MVPA multivoxel pattern analysis OFA occipital face area PACE prospective acquisition correction TR repetition time

Fig. S1. A basis set of 15 face-place pairs used in decoding phase. Each pair was used twice in each condition, once with the face picture set as target and the other time with the place picture set as target. Note: Copyrighted pictures used in the original experiment have been replaced in the above graphic by their non-copyrighted look-alikes. Fig. S2. Graph-based visual saliency algorithm was used to select the face-place pairs. Saliency of the 50/50 hybrid and each of its constituents were Resveratrol observed and only those pairs were selected for which the 50/50 hybrid had an equal number of salient points for the face and place picture. Fig. S3. Stimulus timeline. (A) Example of an attend-face trial in non-feedback condition. (B) Example of an attend-place trial in feedback condition. After cues have been presented, the face-place hybrid image was updated every TR depending on classification result of the preceding TR. Fig. S4. List of all brain regions from which voxels were selected by the MVA-W classifier for training. Only regions that were activated across three or more subjects were used for further analyses. Fig. S5.

For mixed-strain competitions, hatchlings were exposed to an inoc

For mixed-strain competitions, hatchlings were exposed to an inoculum containing an ∼1 : 1 ratio of wild type and mutant. At 48-h postinoculation,

individual squid were homogenized and dilution plated on LBS. The resulting colonies were patched onto LBS with added trimethoprim to determine the ratio of strains in each animal. Inocula were similarly plated and patched to determine the starting ratio. The relative competitiveness index (RCI) was determined by dividing the mutant to wild-type ratio in each animal by the ratio of these strains in the inoculum. The Ibrutinib mean RCI was calculated from log-transformed data. blast searches (Altschul et al., 1990) of the V. fischeri ES114 genome revealed the similarity of ORFs VF1308 and VF1309 to the N and C termini of E. coli FNR, respectively (Fig. 1a). We MAPK Inhibitor Library nmr suspected that a sequencing error had led

to the misannotation of fnr as two genes, and we therefore cloned and sequenced the region spanning VF1308 and VF1309. We found five errors in the genome database, leading to an erroneously predicted truncation of VF1308, which we corrected in GenBank (Mandel et al., 2008). In the revised sequence, VF1308 encodes a protein that is the same length as, and shares 84% identity with, E. coli FNR. This ES114 FNR is identical to the previously deposited V. fischeri MJ1 FNR (accession no. CAE47558). Importantly, the residues necessary for interactions with RNA Molecular motor polymerase (Williams et al., 1997; Lonetto et al., 1998; Blake et al., 2002; Lamberg et al., 2002), 4Fe–4S center assembly (Spiro & Guest, 1988; Kiley & Beinert, 1998), and DNA recognition (Spiro et al., 1990) in E. coli are conserved in V. fischeri FNR. Using TransTermHP (Kingsford et al., 2007), we also found a likely Rho-independent transcriptional terminator downstream of fnr (Fig. 1a and b). Given the 142-bp spacing and strong putative terminator between fnr and VF1310 (Fig. 1b), it seems likely that these are expressed on separate transcripts. Using quantitative RT-PCR, we found that the fnr∷tmpR allele in mutants described

below did not affect the transcript levels for VF1310. We next generated mutants disrupted in the putative fnr in V. fischeri ES114 and MJ1. We did not observe any attenuation of these strains under aerobic growth conditions, consistent with the role of FNR in other bacteria. Escherichia coli fnr mutants do not grow anaerobically with nitrate or fumarate as an electron acceptor (Lambden & Guest, 1976), and we found that V. fischeri fnr mutants were similarly attenuated. Specifically, when grown with minimal medium under anaerobic conditions, ES114 and MJ1 displayed nitrate- or fumarate-dependent growth on a nonfermentable carbon source (glycerol) that was lacking in the fnr mutants (e.g. Fig. 1c).

Surprisingly, commercial sex workers and clients

Surprisingly, commercial sex workers and clients selleckchem of commercial sex workers were not less likely to have their source tested than the rest of the study population. The difference between heterosexual and homosexual subjects could not be explained by differences in frequency of anonymous contacts, as one

might have expected. However, it is possible that the definition of anonymous contacts did not encompass the same realities in the two groups, as many anonymous MSM contacts occurred in bathhouses with truly untraceable contacts. Testing the source also allowed us to detect 11 undiagnosed HIV infections. The HIV prevalence of the source population of unknown HIV status was therefore 3.7%, a proportion 10 times higher than that reported in the general population in Switzerland [27]. When source subjects that were reported to be HIV MEK inhibitor positive were included, the prevalence increased to 24%,

which is consistent with other reports [13,17]. Sixty-two per cent of those for whom information was available were not treated and 69% had a detectable viral load. These data underscore the risk of undiagnosed and untreated HIV infection in the population of source subjects and therefore support the prescription of nPEP in cases of exposure to persons of unknown HIV status belonging to high-risk groups. However, in this study, a significant proportion (58%) of subjects reporting heterosexual contact with an anonymous or a casual partner were prescribed nPEP, although the source was not reported to belong to any risk group for HIV infection. Although this practice is not endorsed by our national guidelines, antiretroviral prophylaxis was provided in these cases because the source

was reported to have multiple sexual partners and believed to be at risk for HIV infection. We observed two seroconversions. Neither was linked to nPEP failure, as infection occurred after ongoing risk behaviour. The fact that one of the two patients was not offered prophylaxis at the time of consultation does not call into question Pregnenolone our policy to withhold nPEP when the source is tested negative. Indeed, fourth-generation tests have recently been shown in percutaneous occupational exposures to detect p24 antigen during acute HIV infection when antibodies are still undetectable [28]. The absence of nPEP failure, however, cannot be considered proof of its efficacy as the sample size was too small to allow assessment of such a rare phenomenon. A major limitation of our study was the high drop-out rate throughout the follow-up period. Overall, 16% of patients for whom nPEP was initiated never came back for assessment of regimen completion and drug toxicity and 49% of all participants never had a second HIV test at 3 months.

However, clinicians must decide whether the attributed benefits a

However, clinicians must decide whether the attributed benefits are clinically significant, considering the costs and potential risks of GH axis treatments. A limitation of this study is the small number of studies available of each GH axis drug class. Disorders of body fat metabolism and associated metabolic buy MK0683 alterations are common in patients infected with HIV [1]. While the definition is not standardized, a diagnosis of HIV-associated lipodystrophy describes metabolic derangements including insulin resistance and changes in lipid metabolism, which result in lipoatrophy (peripheral adipose wasting) and lipohypertrophy (visceral adipose accumulation)

[1]. The pathogenesis of HIV-associated lipodystrophy is multifactorial and includes genetic predisposition, effects of antiretroviral agents, HIV infection itself, and other host factors [2]. Highly active antiretroviral therapy (HAART) is comprised of potent antiretroviral agents, which have dramatically improved clinical outcomes in patients with HIV infection. Unfortunately, HAART is often associated with the

onset, or exacerbation, LDK378 cost of HIV-associated lipodystrophy [1]. The prevalence of HIV-associated lipodystrophy is 4% in untreated patients and 13–40% in patients on HAART [3]. The associated fat redistribution syndrome can lead to negative social, psychological and medical consequences

[4]. Cosmetic changes in body shape may result in decreased compliance with HAART, which can result in increased viral replication and associated morbidities and mortality [4]. The metabolic derangements in HIV-associated lipodystrophy are difficult to reverse. A number of treatments for this condition have been explored, including metformin, thiazolidinediones (TZDs), testosterone and growth hormone (GH) axis drugs. Metformin has been shown to reduce visceral adipose tissue (VAT) mass but accelerates peripheral adipose tissue loss. TZDs and testosterone are not effective in reducing triclocarban VAT. However, some studies have shown that GH axis drugs can both decrease VAT and help to maintain or improve peripheral adipose tissue mass [5]. Although the underlying mechanism for the development of HIV-associated lipodystrophy and related disorders such as metabolic syndrome is not fully understood, evidence suggests that neurohormonal dysregulation plays a role in causing these debilitating conditions [6–8]. GH is a polypeptide hormone secreted episodically by the adenohypophysis that affects protein, carbohydrate and lipid metabolism. There is also evidence that it plays a role in skeletal and visceral growth. Specifically, GH affects the metabolism of fats by causing cells to switch from using carbohydrates for fuel to burning fats for energy.

Thus, it is suspected that augmenting the GH axis in patients wit

Thus, it is suspected that augmenting the GH axis in patients with HIV-associated lipodystrophy results in improved utilization of fat stores and subsequent redistribution of adipose tissue [9]. GH axis drugs investigated for the treatment of HIV-associated lipodystrophy include recombinant growth hormone (GH), growth FK506 cost hormone releasing hormone (GHRH), tesamorelin, also known as growth hormone releasing factor (GHRF), and insulin-like growth factor-1 (IGF-1). There are some concerns with this class of drug. GH, the most studied GH axis drug, costs approximately $52 per milligram, and is estimated to cost approximately US$10 000–US$30 000

per year of treatment [10]. Significant treatment-associated side effects of these drugs include arthralgias, myalgias, peripheral oedema,

insulin resistance and diabetes [11]. Considering the expense and side effects associated with these drugs, it is important to evaluate the evidence regarding the efficacies of these treatments to allow the patient and health care provider to make informed decisions. In the present systematic review, we evaluate randomized controlled trials comparing the effects of GH axis treatments with those of placebo in changing VAT, subcutaneous adipose tissue (SAT) and lean body mass (LBM) in patients with GW-572016 in vitro HIV-associated lipodystrophy. A detailed protocol was written prior to conducting the review. The protocol and documentation of all changes made after construction of the protocol are available upon request. Inclusion criteria were as follows (all were required to be met): (1) the study design must be a randomized controlled trial; (2) study participants were adult patients with HIV-associated lipodystrophy;

(3) the intervention was a GH axis drug (GH, GHRH, tesamorelin or IGF-1); (4) the comparison group was treated with placebo; and (5) the study included one of the primary outcomes. There were no exclusion criteria. Our primary outcomes of interest included changes in VAT mass, SAT mass or LBM. The secondary these outcomes included changes in extremity fat, levels of fasting plasma glucose, high-density lipoprotein (HDL) cholesterol and triglycerides, and waist circumference. Potential harms of treatment were also evaluated. The following databases were searched for studies: OVID MEDLINE (1996 to present; accessed 6 June 2010), The Cochrane Library [Cochrane Central Register of Controlled Trials and Cochrane Database of Randomized Controlled Trials (CENTRAL); accessed 11 October 2009], Web of Science (accessed 11 October 2009), Summons (accessed 13 October 2009), Google Scholar (accessed 11 October 2009) and PubMed (accessed 5 June 2010). Search terms included: HIV, AIDS, growth hormone, Serostim, GH releasing hormone, tesamorelin, IGF-1, HIV-associated lipodystrophy, adipose tissue and body composition. A comprehensive list of all search terms is available upon request.

A traditional defining characteristic of members of the Shewanell

A traditional defining characteristic of members of the Shewanella genus is the inability to use glucose as a substrate for growth. Shewanella spp. isolates, however, have the common ability to use a diverse array of substrates, allowing them to survive in a range of environments (Hau & Gralnick, 2007; Fredrickson et al.,

2008). Members of the Shewanella genus show great flexibility with regard to alteration of growth strategy and metabolism based on the availability of different carbon sources (Tang et al., 2009). In keeping with this flexibility, the traditional view of inability to use glucose has changed as many Shewanella spp. isolates have since been found to use glucose (Bowman et al., 1997; Nogi et al., 1998; Leonardo et al., 1999; Brettar et al., 2002; Gao et al., 2006; Zhao et al., 2006; Xiao et al., 2007; Rodionov et al., 2010). To this end, Shewanella oneidensis selleckchem MR-1, isolated from sediment in Lake Oneida, NY, has not been yet observed to use glucose as a fermentation substrate, under short-term growth experiments without initial glucose exposure in a rich growth medium (Myers & Nealson, 1988; Venkateswaran et al., 1999; Rodionov et al., 2010). In microbial fuel cells (MFCs) with complex

growth media, however, S. oneidensis Tamoxifen in vitro has been found to generate current upon extended glucose addition (Biffinger et al., 2008, 2009). This response to glucose addition was slow, suggesting that S. oneidensis may be able to use glucose, given ample time to induce the appropriate genetic mechanisms, for a mutator population to proliferate (Chao & Cox, 1983; Giraud et al., 2001a, b) and/or

develop a ‘growth advantage in stationary phase’ (GASP) mutant (Finkel & Kolter, 1999). Mutator bacteria contain mutations that inactivate mutation-avoidance genes yielding higher spontaneous mutation rates, which in turn yield an increased evolutionary pace (Chao & Cox, 1983; Mao et al., 1997; Giraud et al., 2001a, b). GASP refers to the genetic alterations (not physiological adaptations) that occur in cells incubated in long-term batch cultures that confer a competitive advantage to these cells over younger ‘naïve’ cultures (Finkel, 2006). Given that the S. oneidensis genome suggests the ability to use glucose as a Liothyronine Sodium fermentation substrate via the Entner–Doudoroff pathway, the current study seeks to show whether S. oneidensis can indeed utilize glucose as a sole carbon source given an initial glucose exposure, as suggested by previous MFC studies (Biffinger et al., 2008, 2009). Shewanella oneidensis MR-1 (Venkateswaran et al., 1999), obtained from the American Type Culture Collection (ATCC#700550) and stored at −80 °C, was grown up in Luria–Bertani (LB) broth for 24 h, shaking (100 r.p.m.) at 25 °C. From the LB culture, S. oneidensis MR-1 was serially passed every 24 h for 96 h into LB broth amended with 10 mM glucose.

More recently, C vulgaris NJ-7, a strain isolated from the Antar

More recently, C. vulgaris NJ-7, a strain isolated from the Antarctic, was used to investigate the adaptation of eukaryotic microbes to permanently cold environments (Hu et al., 2008; Li et al., 2009). The strain NJ-7 possesses the same 18S rRNA gene sequence as that of UTEX259, a strain isolated

from the temperate region, but shows a significantly intensified freezing tolerance (5- to  1000-fold higher viability) than the temperate strain. Comparative studies of the two C. vulgaris strains provide opportunities to understand how intra-species learn more evolution is undertaken in eukaryotic microbes to adapt to the Antarctic or other extreme environments. HIC6 is a group-3 late embryogenesis abundant (LEA) protein found in C. vulgaris. Together with HIC12, it was first identified by 2D-HPLC and SDS-PAGE to be hardening (cold treatment)-induced in the strain C-27 (Honjoh et al., 1995). Its cDNA was also identified by differential screening of a cDNA library (Joh et al., 1995) or suppression subtractive hybridization (Machida et al., 2008). LEA Rapamycin mouse proteins were initially found at the late stage of embryogenesis in cotton (Galau et al., 1986) and were subsequently found in algae (such as C. vulgaris), cyanobacteria (Close & Lammers, 1993), nematodes (Browne

et al., 2002) and fungi (Abba’ et al., 2006). The proteins can be divided into different groups on the basis of similarities in amino acid sequences (Colmenero-Flores et al., 1997; Cuming, 2005; Battaglia et al., 2008). Like many other LEA proteins, HIC6 remained soluble under boiling conditions and showed in vitro cryoprotective activities on lactate dehydrogenase (LDH) (Honjoh et al., 2000). Overexpression of HIC6 in plant or yeast could enhance their freezing tolerance (Honjoh et al., 1999, 2001), and

in the transgenic plant, HIC6 was localized to mitochondria (Honjoh et al., 2001). In strains NJ-7 and UTEX259, the encoding gene hiC6 was also induced upon exposure to cold, and the expression was intensified in strain NJ-7 in comparison with UTEX259 (Li et al., 2009). These results suggest that the enhanced expression of hiC6 is probably involved in the development of freezing tolerance in C. vulgaris. The intensified expression of hiC6 in NJ-7 could be due to gene duplication, increased transcription or post-transcriptional regulation. Obatoclax Mesylate (GX15-070) In our previous study, only one hiC6 gene was identified in each of the two Chlorella strains, NJ-7 and UTEX259 (Li et al., 2009). In the present study, however, sequencing of that chromosomal region revealed that multiple hiC6 genes are organized in tandem in both strains. The tandem-arrayed genes encode different HIC6 isoforms and are differentially expressed. Chlorella vulgaris strains were grown in BG11 (Stanier et al., 1971) in the light of 50 μE m−2 s−1 at 20 °C with aeration. Cells grown at 20 °C were cooled to 4 °C in a water bath and transferred to a 4 °C refrigerator with aeration and illumination (50 μE m−2 s−1) for different periods of time.

1b and c) The noncovalent inhibitors including benzamidine, leup

1b and c). The noncovalent inhibitors including benzamidine, leupeptin and

nafamostat mesylate also showed weak inhibition of HsaD (Fig. 1b) compared to PMSF and DCI. MGL like HsaD catalyses the turnover of highly hydrophobic substrates: as such the inhibitors LY294002 mw that have been identified tend to be insoluble (e.g. pristimerin and NAM). Although pristimerin is the most active noncovalent inhibitor tested (35% inhibition at 50 μM – Fig. 2a), further investigation was hampered by its poor aqueous solubility under conditions that are required for HsaD to remain active. NAM and JZL184 are covalent inhibitors: JZL184 like DCI and PMSF modifies the catalytic serine of MGL (Long et al., 2009), while NAM modifies a cysteine in the active site of MGL (Saario et al., 2005). Consistent http://www.selleckchem.com/products/Staurosporine.html with the lack of a cysteine residue in the active site of HsaD, NAM does not significantly inhibit HsaD (Fig. 2a). JZL 184 proved a better inhibitor (Fig. 2a) but was difficult to work with due to its hydrophobic nature and hence poor solubility. A series of specific acetylcholinesterase inhibitors were tested for inhibition of HsaD (Fig. 2b). These included eserine, edrophonium, tacrine, neostigmine, pyridostigmine and trichlorfon. After incubation with HsaD, trichlorfon inhibited poorly. Eserine and neostigmine show better inhibition, but still not as strong as was observed with DCI (c. 30% inhibition at 1 mM). The other

acetylcholinesterase inhibitors did not significantly inhibit HsaD. Two mechanisms have been proposed for the hydrolysis of substrates by MCP hydrolases. The first is based on the mechanism known to occur in serine proteases and proceeds via an acyl enzyme and tetrahedral intermediate (Ruzzini et al., 2012). The second requires a keto-enol tautomerization resulting in a gem-diol intermediate (Horsman et al., 2007). Recent mutagenesis experiments combined with structural studies resulted in trapping of the acyl enzyme intermediate of HOPDA hydrolysis, by another member of the C-C bond hydrolase family, BphD (Ruzzini et al., 2012) strongly supporting the first mechanism. Inhibition by PMSF and

DCI is also consistent with this mechanism as PMSF and DCI act as tetrahedral and acyl enzyme intermediate analogues, respectively, when they modify the active Oxalosuccinic acid site serine. The most successful inhibitors were those that covalently modify HsaD (e.g. DCI). The primary issue with DCI and other covalent inhibitors tends to be their broad specificity profile making them poor starting points for inhibitor design. To help understand the specificity observed among the covalent inhibitors, the structure of HsaD modified with PMSF was solved (Fig. 3). Although density was observed for the sulphonate group covalently linked to Ser114, there was insufficient density to accommodate the phenylmethyl group of PMSF. A lack of electron density for PMSF in the structure with HsaD might suggest that PMSF acts reversibly.

1b and c) The noncovalent inhibitors including benzamidine, leup

1b and c). The noncovalent inhibitors including benzamidine, leupeptin and

nafamostat mesylate also showed weak inhibition of HsaD (Fig. 1b) compared to PMSF and DCI. MGL like HsaD catalyses the turnover of highly hydrophobic substrates: as such the inhibitors find protocol that have been identified tend to be insoluble (e.g. pristimerin and NAM). Although pristimerin is the most active noncovalent inhibitor tested (35% inhibition at 50 μM – Fig. 2a), further investigation was hampered by its poor aqueous solubility under conditions that are required for HsaD to remain active. NAM and JZL184 are covalent inhibitors: JZL184 like DCI and PMSF modifies the catalytic serine of MGL (Long et al., 2009), while NAM modifies a cysteine in the active site of MGL (Saario et al., 2005). Consistent Selleck LY2157299 with the lack of a cysteine residue in the active site of HsaD, NAM does not significantly inhibit HsaD (Fig. 2a). JZL 184 proved a better inhibitor (Fig. 2a) but was difficult to work with due to its hydrophobic nature and hence poor solubility. A series of specific acetylcholinesterase inhibitors were tested for inhibition of HsaD (Fig. 2b). These included eserine, edrophonium, tacrine, neostigmine, pyridostigmine and trichlorfon. After incubation with HsaD, trichlorfon inhibited poorly. Eserine and neostigmine show better inhibition, but still not as strong as was observed with DCI (c. 30% inhibition at 1 mM). The other

acetylcholinesterase inhibitors did not significantly inhibit HsaD. Two mechanisms have been proposed for the hydrolysis of substrates by MCP hydrolases. The first is based on the mechanism known to occur in serine proteases and proceeds via an acyl enzyme and tetrahedral intermediate (Ruzzini et al., 2012). The second requires a keto-enol tautomerization resulting in a gem-diol intermediate (Horsman et al., 2007). Recent mutagenesis experiments combined with structural studies resulted in trapping of the acyl enzyme intermediate of HOPDA hydrolysis, by another member of the C-C bond hydrolase family, BphD (Ruzzini et al., 2012) strongly supporting the first mechanism. Inhibition by PMSF and

DCI is also consistent with this mechanism as PMSF and DCI act as tetrahedral and acyl enzyme intermediate analogues, respectively, when they modify the active Resminostat site serine. The most successful inhibitors were those that covalently modify HsaD (e.g. DCI). The primary issue with DCI and other covalent inhibitors tends to be their broad specificity profile making them poor starting points for inhibitor design. To help understand the specificity observed among the covalent inhibitors, the structure of HsaD modified with PMSF was solved (Fig. 3). Although density was observed for the sulphonate group covalently linked to Ser114, there was insufficient density to accommodate the phenylmethyl group of PMSF. A lack of electron density for PMSF in the structure with HsaD might suggest that PMSF acts reversibly.