To assess the influence of the history and examination findings on antibiotic prescribing where LRTI is the principal diagnosis, and to explore the attitudes towards antibiotic prescribing through an understanding of the clinician and patient experience. Although hospitalised patients are unlikely to have as big an influence on choice of therapy as in general practice this adjunct to the main study will seek to elicit

the impact of the condition on daily life, the choice of antibiotic treatment versus no treatment and the potential problems of antibiotic treatment from the patient’s perspective. A mixed methodology study of adult hospitalised patients, with an observational cohort for the quantitative arm and a phenomenological study for the qualitative arm of the research. Data will selleckchem Metformin in vitro be collected from patients’ medical notes using a coding matrix developed as part of a pilot study. Data collected will include demographic details, symptoms and signs, diagnosis, diagnostic tests and results. Doctors will be invited to participate in interviews to discuss the reasons for

prescribing antibiotics in respiratory tract infection. A purposive sample of patients will be selected based on demographics and treatment received to participate in a short interview seeking their views on treatment versus no treatment in LRTI. Admissions data has been collected on 112 patients thus far with ages ranging Methamphetamine from 20 to 95, 64 males and 48 females. Preliminary quantitative data indicate that the diagnosis of LRTI and prescription of antibiotics is made on the recorded presence of a very small number of symptoms and signs, with 93% having shortness of breath, 78% having a respiratory rate >20/minute and 74% having purulent sputum. All patients had at least one X-ray. Interpretation of the films, prior to starting antibiotics, was by the admitting team. Laboratory investigations performed included blood culture in 20% to CRP in 39% of patients. 25% had a working diagnosis of pneumonia whilst 100% of patients

received one or more antibiotics. Ethical committee approval was received and all participants gave informed consent. The results indicate that the diagnosis of LRTI is made using very few recorded criteria. Easy access to radiology and pathology in hospital can assist the diagnosis and should ensure appropriate prescribing of antibiotics. However, whilst 100% of patients received an X-ray, pathology was less utilised. Pneumonia remains a disease with considerable morbidity and mortality worldwide and treatment with antibiotics is generally justified. However, with increasing concerns over antibiotic resistance, the rise in the incidence of healthcare-acquired infections and financial pressures on the medicines’ budget their use should be targeted at those for whom they are appropriate and whose benefit will be greatest.

, 2007). To compare relative expression levels of sas016 in wild type and mutant strains, overnight cultures were diluted to OD 0.05 in prewarmed LB broth and cultures grown to OD 1.5, except for the LCP triple mutant

that was sampled at OD 0.5 because of its severe growth defect. Uninduced culture samples were collected, and the remainder of the culture was induced with oxacillin (10 μg mL−1) for 30 min before induced samples were collected. Total RNA was extracted as described by Cheung et al. (1994). RNA samples (9 μg) were separated in a 1.5% agarose-20 mM guanidine thiocyanate gel in 1× TBE buffer (Goda & Minton, 1995). The sas016 digoxigenin (DIG)-labelled probe was amplified using the PCR DIG Probe synthesis kit (Roche) as previously described (Dengler et al., 2011). The transcriptional start site of sas016 was determined by primer extension, as previously described (McCallum et al., 2007), using primer Selleck EPZ015666 SAS016.PErev (Table 1) and 20 μg of RNA Selleckchem INK-128 harvested from a culture of S. aureus COL that had been grown to OD 0.5 and induced with 10 μg mL−1 of teicoplanin for 30 min. The promoter region of the vraSR operon was PCR amplified from S. aureus strain COL using primer pair vra.lucF and vra.lucR (Table 1). The PCR product was digested with Asp718 and NcoI and ligated directly upstream of the promoterless luciferase

(luc+) gene in the vector pSP-luc+ (Promega). Fragments containing the resulting promoter-luc+ translational fusions were then excised with Asp718 and EcoRI and cloned into the Escherichia coli – S. aureus shuttle vector pBUS1 (Table 1). The fusion plasmids pvrap-luc+ Montelukast Sodium and psas016p-luc+ (McCallum et al., 2011) were then electroporated into S. aureus RN4220, re-isolated and electroporated into S. aureus SA113, SA113ΔtarO, MSSA1112 and all LCP and VraR/LCP mutants. Predicted VraR-binding sites of luciferase fusion constructs were disrupted by amplifying each promoter as two fragments, using primers listed in Table 1. Complementary fragments were digested and ligated together, to create recombinant promoters in which 6–18-bp regions were replaced by restriction

sites. Promoters were then fused to the luciferase gene as described above, and the resulting plasmids were electroporated into RN4220. To measure luciferase activities, cultures were grown from overnight cultures inoculated to an OD 0.05 in prewarmed LB broth containing tetracycline. One-millilitre culture samples were harvested by centrifugation, and the pellets frozen at −20 °C. To determine relative light units (RLU), pellets were thawed briefly and resuspended in PBS (pH 7.4) to an OD of either 10 or 1, depending on induction levels. Aliquots of the cell suspensions were then mixed with equal aliquots of Luciferase Assay System substrate (Promega), and luminescence was measured for 15 s after a delay of 3 s on a Turner Designs TD-20/20 luminometer (Promega) as previously described (Dengler et al., 2011).

Thus, reduced mirror activity was not a direct consequence of a larger EMG burst from

the trained hemisphere causing increased IHI onto the non-trained hemisphere (Hinder et al., 2010). We can also exclude the possibility that the effects are related to attention as they were not influenced by the presence of feedback, which potentially influences the attentional resources. Finally, there was no correlation between the practice-related changes of EMG mirroring check details and corticospinal excitability of the trained hemisphere. We conclude that reduction of EMG mirroring is a process that is separate from improving performance of the trained hand and practice-related corticospinal plasticity of the trained hemisphere. As stated above, although there was no overall change of s- and l-IHI after training, the individual maximal level of s-IHI, but not the individual maximal level of l-IHI, prior to training correlated with the reduction in mirror activity that occurred

during training. Thus, the present results suggest that the motor training-related effects on the EMG mirroring are specific to one interhemispheric motor pathway, mediated by a population of GABAergic interneurons (Irlbacher et al., 2007), which are thought to play a predominant role in the suppression of EMG mirroring during fast finger movements (Duque et al., 2007; Hübers et al., 2008). We speculate that the excitability of s-IHI measured at rest is a measure of ‘resource’, that is, it gives an indication Dabrafenib of what level of IHI is available to the system to employ during voluntary movement. In fact, because IHI is directly related to the structural measures (magnetic resonance imaging fractional anisotropy) of the anatomy of the mid-portion of the corpus callosum (Wahl et al., 2007; Koerte et al., 2009), it may give an indication of the physical limits of IHI. Thus, individuals with greater s-IHI at rest will have a greater potential for controlling EMG mirror activity during training of intentional movement. In this scheme, motor practice does not reduce EMG mirroring by increasing the sensitivity of IHI. Instead,

EMG mirroring may decline because the motor command is better targeted at the task being performed. The more ‘resource’ that there Terminal deoxynucleotidyl transferase is available in s-IHI, the more efficiently this focussing can reduce EMG mirroring activity. Although recent studies have shown that there are similar structure–function relationships when examining GABA-A-mediated IHI, i.e. s-IHI (Wahl et al., 2007), as those found with l-IHI (Fling & Seidler, 2012), the present results confirm that only s-IHI has a functional role in the suppression of the EMG mirroring during fast finger movements (Duque et al., 2007; Cincotta & Ziemann, 2008; Hübers et al., 2008). In previous studies it has been shown that IHI from the trained to the untrained motor cortex can show plastic changes, mainly seen as a reduction of IHI (Shim et al., 2005; Perez et al., 2007; Camus et al., 2009; Hortobágyi et al.

putida PCL1445 (data not shown). The stability of the plasmids and the transposon integration was

tested by subculturing in nonselective media (without antibiotic selection pressure) for approximately 30 generations. Samples of the subcultures were plated and colonies were screened for the expression of mcherry by fluorescence microscopy. Strain PCL1481 carrying miniTn7∷mcherry did not show any loss of integration. No loss of plasmid was observed for PCL1479 carrying pMP7604, whereas 3% of the colonies of strain PCL1480 carrying pMP7605 had lost fluorescence at day 3 (data not shown). A qualitative and quantitative analysis for mCherry production in P. putida PCL1445 tagged with pMP7604, pMP7605 and pMP7607 was performed in order to evaluate the resulting brightness of the different AZD6244 solubility dmso constructs. Cells of overnight cultures were visualized using fluorescence and light microscopy (Fig. 3a) and fluorescence was quantified using fluorometry (Fig. 2b). mcherry expression was detected at the single-cell level for all tagged strains. Microscopic and fluorometric analyses showed that strain PCL1480 (harboring pMP7605) produced the highest amount of mCherry and strain PCL1481 (containing miniTn7-mcherry)

produced the lowest amount (Fig. 3a and b). The strains PCL1479, Bcl-2 inhibitor PCL1480 and PCL1481 produced mCherry in a ratio of 15 : 95 : 1, respectively. No significant fluorescence was detected for P. putida PCL1445 cells and strains PCL1477 and PCL1478 containing the cloning vectors pME6031 and pBBR1MCS-5 (Fig. 2b). To evaluate the applicability of the mCherry marker vectors for tagging Gram-negative bacteria, several other Gram-negative spp., such as P. fluorescens WCS365 (an efficient root colonizer), P. aeruginosa PAO1 (a model strain for cystic fibrosis research)

and E. tarda FL6-60 (a fish pathogen and model for zebrafish immunology), were transformed with pMP7604 and pMP7605. This yielded PCL1700, PCL1701, PCA0241, PCA0242, PCA0239 and PCA0240, respectively. Fluorescence microscopy analysis showed the production of mCherry for all transformed strains (data not shown). Single colonies were isolated and overnight cultures were grown for quantitative analysis of mCherry production and comparison with P. putida PCL1445 (Fig. 4). Strains containing pMP7605 showed the highest mCherry Bay 11-7085 production. Comparable mCherry production levels were observed among the four strains tested, except for the one carrying pMP7605, which showed a lower level of expression in E. tarda FL6-60. To analyze the applicability of the mcherry-expressing constructs pMP7604, pMP7605 and pMP7607 in established test systems, which are not suitable for efficient application of antibiotic pressure, P. putida PCL1445-tagged strains were allowed to form biofilms on glass (in vitro biofilm assay) and on tomato roots (in vivo assay used to study root colonization). Using CLSM, the tagged strains were visualized at the single-cell level in both assays (Fig.

2). CDD analysis (Marchler-Bauer et al., 2011) (data not shown) revealed that the predicted gene product of each contains the conserved PhaC N-terminus domain (pfam07167) and the expected α/β hydrolase fold (pfam00561) (Rehm, 2003). Phylogenetic analysis, presented in the Supporting Information (Figs S1 and S2), reinforced that these genes are homologous to, but substantially JNK inhibitor different

from, known PHA synthesis genes. In clone pCX92, phaC is within a cluster of genes with an organization similar to a segment of the genome of Novosphingobium aromaticivorans, a member of the Alphaproteobacteria. The %GC of the pCX92 sequence, at 65.7, is very similar to the %GC of the corresponding region of the N. aromaticivorans genome, at 64.8. For each of the genes, the corresponding

N. aromaticivorans gene is the highest match, ranging from 51% to 89% amino acid sequence identity, with the phaC exhibiting 66% amino acid sequence identity. In an arrangement similar to that found in the N. aromaticivorans genome, this clone also contains a putative phasin-encoding gene immediately adjacent to the phaC gene. The clone does not contain any other polyhydroxyalkanaote cycle genes, but this is not unusual, as a broad diversity in genomic organization of polyhydroxyalkanaote synthesis genes has been long recognized (Rehm & Steinbüchel, 1999). The sequence of the pCX9M4 subclone pMS2 revealed the phaC gene to share 56% amino acid sequence identity with a phaC gene from Thauera sp. MZ1T, a member of the Betaproteobacteria, Obeticholic Acid mw and to be adjacent to a phaB gene. The %GC of

the pMS2 sequence, at Rho 66.5, is very similar to the %GC of the corresponding region of the Thauera sp. MZ1T genome, at 66.0%. Curiously, maximum-likelihood phylogenetic analysis (Figs S1 and S2) clusters the pMS2 phaC sequence with the MZ1T phaC sequence at the amino acid level only, not at the DNA level, despite the very similar %GC. Because the complete sequence of pCX9M4 has not yet been determined, we do not know whether additional polyhydroxyalkanaote cycle genes are present on the clone, but the MZ1T genome has a phaR repressor gene further downstream of phaC-phaB. The sequence of the pCX9M5 subclone pMS3 indicated a phaC gene with 61% amino acid sequence identity to the well-studied phaC gene of Cupriavidus necator H16, also from the Betaproteobacteria. The best-matching genomic fragment, however, was with another member of the Betaproteobacteria, Burkholderia sp. 383, despite differences in %GC, 59.4 for pMS3 compared with 66.9 for Burkholderia sp. 383. The phaC gene is located adjacent to a phaA. Like pCX9M4, the complete sequence of pCX9M5 has not yet been completed, and so we do not know whether other polyhydroxyalkanaote cycle genes are present on this clone. However, Burkholderia sp. 383 has the typical genomic organization of a class I operon (phaCABR).

“
“Trichomonas vaginalis is a parasite that resides in the human urogenital tract and causes trichomonosis, the most prevalent nonviral sexually transmitted disease. Nucleoside triphosphate diphosphohydrolase (NTPDase), which hydrolyzes extracellular di- and triphosphate nucleotides, and ecto-5′-nucleotidase, which hydrolyzes AMP, have been characterized in T. vaginalis. The aim of this study was to characterize the adenosine Rapamycin mouse deaminase (ADA) activity in intact trophozoites of T. vaginalis. A strong inhibition in adenosine deamination was observed in the presence of calcium and magnesium, which was prevented by

EDTA. The apparent KM value for adenosine was 1.13 ± 0.07 mM. The calculated Vmax was 2.61 ± 0.054 nmol NH3 min−1 mg−1 protein. Adenosine deamination was inhibited in the presence of erythro-9-(2-hydroxy-3-nonyl)adenine. Semi-quantitative reverse transcriptase-PCR experiments were performed and both ADA-related genes ada(125) and ada(231) mRNA were expressed, although ada(231) in higher quantity when compared with the ada(125) : α-tubulin ratio. Furthermore, a phylogenetic analysis showed that the T. vaginalis sequences formed a clade with Entamoeba histolytica and Dictyostelium discoideum sequences, and it strongly suggests homologous functions in the T. vaginalis genome. The presence selleck of ADA activity in T. vaginalis may be important to modulate the

adenosine/inosine levels during infection and, consequently, to

maintain the anti-inflammatory properties through different nucleoside-signalling mechanisms. Trichomonas vaginalis is a protozoan parasite that causes trichomonosis, the most prevalent nonviral sexually transmitted selleck chemical disease worldwide (WHO, 2001). In women, the infection is clinically characterized by vaginitis and cervicitis (Petrin et al., 1998; Lehker & Alderete, 2000). The pathogen has been associated with serious health consequences including adverse pregnancy outcomes (Klebanoff et al., 2001), infertility (Grodstein et al., 1993), predisposition to cervical cancer (Viikki et al., 2000) and pelvic inflammatory disease (Cherpes et al., 2006), and it is a cofactor in HIV transmission and acquisition (Sorvillo et al., 2001; Van Der Pol et al., 2008). At the infection sites, tissue stress or injury takes place and intracellular ATP can be released into the extracellular environment. Extracellular nucleotides such as ATP play a role as danger-associated molecular patterns (DAMPs) or ‘alarmins’ by acting as signalling molecules that contribute to inflammation and immune responses (Hanley et al., 2004; Bours et al., 2006). The crucial factors in purinergic signalling are the stimulation of nucleotide release, their metabolism by enzymes acting in an extracellular manner and the presence of receptors that selectively bind the resulting products and mediate signal transduction (Gounaris & Selkirk, 2005).

(2008). The Antarctic continent has been frequently cited as a pristine place, with a rather limited diversity of plants and animals, but with a highly diverse microbial community (Tindall, 2004). In particular, it Selleckchem Panobinostat was reported that aquatic environments (sea, sea ice, lakes from freshwater

to highly saline) were more diverse compared with soils. However, recent applications of molecular methods have revealed a very wide diversity of microbial taxa in soil, many of which are uncultured and taxonomically unique (Cary et al., 2010; Margesin & Miteva, 2010). Thus, this continent could be considered to be of great importance for several reasons, among them because it might be regarded as a reservoir for novel genetic resources that ALK inhibitor could be of use in the development of new biotechnological products. In addition, Antarctica might be considered a natural laboratory to understand the genetic and structural basis of adaptation of eukaryotic and prokaryotic cells to extreme conditions. This review presents recent

information about the genomic elements that have been found to act in the evolution of the Antarctic prokaryotic genomes and their potential for biotechnological exploitation. Currently, it is accepted that the notion that the Antarctic continent is a pristine environment is misleading because of the input of airborne microorganisms and the anthropogenic transport and dissemination of microorganisms, as an inevitable consequence of human presence and activity. These ‘alien’ microorganisms

do indeed influence the microbial why diversity, giving insight into the complexity of the balance between evolution, extinction, and colonization of microorganisms in this extreme environment (Vincent, 2000; Pearce et al., 2009; Cowan et al., 2011). The continent is continuously seeded by nonindigenous microorganisms including mesophilic species that, although they will probably not establish viable populations, they contribute to the environmental pool of DNA available through one of the major forces in the evolution of the prokaryotic genome, horizontal gene transfer (HGT). In addition to the intromission of ‘alien’ microorganisms, climate changes like global warming strongly affect microbial Antarctic communities. Yergeau et al. (2012) showed an increase in the abundance of fungi and bacteria and in the ratio of Alphaproteobacteria to Acidobacteria in response to experimental field warming, which might result in an increase in soil respiration. On the other hand, Jung et al. (2011) reported diminished fungal and archaeal communities in response to warming temperatures. But, whether there is an increase or decline in a group of microorganisms, the shift in Antarctic microbial communities is not in doubt.

(2008). The Antarctic continent has been frequently cited as a pristine place, with a rather limited diversity of plants and animals, but with a highly diverse microbial community (Tindall, 2004). In particular, it Obeticholic Acid supplier was reported that aquatic environments (sea, sea ice, lakes from freshwater

to highly saline) were more diverse compared with soils. However, recent applications of molecular methods have revealed a very wide diversity of microbial taxa in soil, many of which are uncultured and taxonomically unique (Cary et al., 2010; Margesin & Miteva, 2010). Thus, this continent could be considered to be of great importance for several reasons, among them because it might be regarded as a reservoir for novel genetic resources that www.selleckchem.com/products/epz015666.html could be of use in the development of new biotechnological products. In addition, Antarctica might be considered a natural laboratory to understand the genetic and structural basis of adaptation of eukaryotic and prokaryotic cells to extreme conditions. This review presents recent

information about the genomic elements that have been found to act in the evolution of the Antarctic prokaryotic genomes and their potential for biotechnological exploitation. Currently, it is accepted that the notion that the Antarctic continent is a pristine environment is misleading because of the input of airborne microorganisms and the anthropogenic transport and dissemination of microorganisms, as an inevitable consequence of human presence and activity. These ‘alien’ microorganisms

do indeed influence the microbial click here diversity, giving insight into the complexity of the balance between evolution, extinction, and colonization of microorganisms in this extreme environment (Vincent, 2000; Pearce et al., 2009; Cowan et al., 2011). The continent is continuously seeded by nonindigenous microorganisms including mesophilic species that, although they will probably not establish viable populations, they contribute to the environmental pool of DNA available through one of the major forces in the evolution of the prokaryotic genome, horizontal gene transfer (HGT). In addition to the intromission of ‘alien’ microorganisms, climate changes like global warming strongly affect microbial Antarctic communities. Yergeau et al. (2012) showed an increase in the abundance of fungi and bacteria and in the ratio of Alphaproteobacteria to Acidobacteria in response to experimental field warming, which might result in an increase in soil respiration. On the other hand, Jung et al. (2011) reported diminished fungal and archaeal communities in response to warming temperatures. But, whether there is an increase or decline in a group of microorganisms, the shift in Antarctic microbial communities is not in doubt.

, 2002). PCR products were electrophoresed in a 1% agarose gel and purified with the kit GenElute PCR Clean-up (Sigma) following the manufacturer’s instructions. The purified products were cloned in pGEM-T

Easy Vector System II kit (Promega) or directly used for sequencing. Sequencing was accomplished using the kit BigDye Terminator v3.1 Cycle Sequencing (Applied Biosystems) and an ABI Prism 3130 DNA Sequencer (Applied Biosystems). The sequences were analyzed using chromaslite v2.01 and seqmanii (DNASTAR) programs and subjected to blast searches to retrieve the most closely related sequences. The presence of tRNA genes was determined using tRNAscan-SE 1.21 software (Lowe & Eddy, 1997). Previously reported 16S rRNA gene and ISR sequences from T. soleae and related species, retrieved from GenBank database (http://www.ncbi.nlm.nih.gov/genbank/) and those obtained in this study, were aligned by using the program clustalw (http://www.ebi.ac.uk/Tools/msa/clustalw2/) learn more and examined for areas of similarity and variability between different species and strains. On the basis of the alignments, two variable regions were chosen and a pair of primers was designed by using the primer3 program BEZ235 research buy (http://frodo.wi.mit.edu/; Rozen & Skaletsky, 2000). Primers were synthesized by Thermo Scientific (Ulm, Germany). The PCR amplifications were carried out using the commercial

kit RedTaq ReadyMix (Sigma), which included all necessary reagents except the primers and DNA template. The PCR mixture consisted of reaction buffer (10 mmol L−1 Tris–HCl pH 8.3, 50 mmol L−1 KCl, 1.5 mmol L−1 MgCl2), 200 μmol L−1 triclocarban of each dNTP, 200 nmol L−1 of each primer,

3 U of Taq DNA polymerase, template DNA, and double-distilled water up to a final volume of 50 μL. The amplification was performed in a Mastercycler gradient (Eppendorf) as follows: an initial denaturation at 94 °C for 5 min followed by 45 amplification cycles (denaturation at 94 °C for 1 min, annealing at 57 °C for 45 s, and extension at 72 °C for 1 min), and a final elongation at 72 °C for 5 min. DNA from strain T. soleae a47 was included as a positive control and distilled water as a negative control. PCR products were electrophoresed on a 1% agarose TBE gel stained with SYBR Safe DNA Gel Stain (Invitrogen); a 1-kb DNA ladder (Biotools) was included as a molecular weight marker. To test the specificity of the primers in the PCR procedure, nine T. soleae strains, isolated from three different hosts and including the type strain, and 81 strains of other species, most of them taxonomically and/or ecologically related, were used as positive and negative controls, respectively (Table 1). For PCR amplification, 100 ng DNA template was used for each strain. The detection limit was evaluated using 10-fold serially diluted DNA, isolated from strain T. soleae a47, over the range 100 ng to 100 fg. Large amounts of DNA (0.5–3 μg) were also assayed.

Similar to the reactivity observed for mAbs, most polyclonal antisera

showed a lower reactivity with LaiMut, except some antisera against members of serogroups Icterohaemorrhagiae, Canicola, and Sarmin, which were increased (Table 1). The reaction of the mAbs and polyclonal antisera suggested a substantial change in the profile of the immunoreactive epitopes present on the LaiMut lipopolysaccharide as compared with the parent LaiWT strain. Proteinase K-treated whole-cell lysates of LaiWT and LaiMut were resolved by discontinuous SDS-PAGE and stained. The profiles of both LaiWT and LaiMut were similar, with the exception of a reproducible reduction in the molecular mass of the upper band in the lipopolysaccharide

profile find more GDC-0941 datasheet of LaiMut by approximately 3 kDa in comparison with the wild-type strain (Fig. 1). The reactivity of the F70C7 mAb with LaiMut and LaiWT was evaluated by Western blot analysis. The mAb F70C7 reacted with LaiWT lipopolysaccharide, showing a smear over a range corresponding to 22–60 kDa, a profile consistent with the previously reported reactivity of other anti-lipopolysaccharide mAbs (Jost et al., 1988) (Fig. 2). By contrast, no reaction with the LaiMut lipopolysaccharide was detected; this result is consistent with the low MAT titre measured for F70C7 against LaiMut (Fig. 2). A series of oligonucleotide primers for sequencing the lipopolysaccharide biosynthesis locus was designed using the L. interrogans serovar Lai genome sequence as a reference (Ren et al., 2003). The sequence for both LaiMut and LaiWT was determined for the coding regions LA1626–LA1667 (chromosome 1: bases 1 621 069–1 667 280). In this region, only a Farnesyltransferase single nucleotide difference was identified between the LaiMut and the LaiWT sequence; with reference to the Lai genome sequence, the difference was a T to A base change at base 1 645 132 (Fig.

3). This change is located within LA1647, a 1263 base ORF encoding a putative undecaprenyl-galactosyltransferase. The single base change resulted in an internal inframe stop at amino acid 135 of 420. This report describes a leptospiral lipopolysaccharide mutant from serovar Lai. The approach of using a mAb directed at a determinant found on the lipopolysaccharide to select for escape mutant differs from previous studies that have used polyclonal sera recognizing multiple epitopes for selection. The serological characterization of the LaiMut strain revealed that its mAb-binding profile had altered substantially from the parent strain, with several mAbs no longer able to bind. Furthermore, there was a substantial decrease in MAT titres to the polyclonal antisera not only against the same serogroup but also with cross-reactive antisera against related serogroups (Table 1).