We recommend that all patients with AIDS-defining malignancies should start HAART (level

of evidence 1B) [13]. We suggest that all patients with non-AIDS-defining malignancies who are due to start chemotherapy or radiotherapy should be started on HAART unless contraindicated (level of evidence 2C) [13]. This is based on the well-documented decline in CD4 cell counts associated with chemotherapy and radiotherapy. Although guidelines suggest initiation of prophylaxis against opportunistic infections based on CD4 cell count, this differs in those with malignancies due to the possible profound immunosuppression associated with chemotherapy and radiotherapy. Prophylaxis against Pneumocystis jirovecii pneumonia (PCP) is recommended for those who have a CD4 count less than 200 cells/μL (level of evidence 1A) and should be considered Romidepsin research buy at higher levels in all patients starting chemotherapy

or radiotherapy (GPP) [14]. Chemotherapy and radiotherapy are associated with profound falls in CD4 cell counts even in patients on HAART and the degree of decline in CD4 cell count may be unpredictable [1–3]. The treatment of choice is cotrimoxazole, which may have additional benefits Selleckchem Sorafenib in reducing the incidence of bacterial infections (respiratory, gastrointestinal especially salmonella and possibly CNS infections) [15–18] and toxoplasmosis [19,20]. Alternative prophylaxis should be with dapsone or pentamidine via nebuliser. Prophylaxis against MAC is recommended for individuals with a CD4 cell count less than 50 cells/μL (level of evidence 1B) [14]. Individuals who have or are at risk of a CD4 cell count falling below this level should be considered for MAC prophylaxis. The treatment Thymidylate synthase of choice is azithromycin 1.25 g once per week or clarithromycin with rifabutin being considered as an alternative [21–24]. People living with HIV who have low CD4 cell counts are at risk of fungal infections, most commonly oral and oesophageal candida and cryptococcosis; whilst those with prolonged very low CD4 cell counts are also

at risk of pulmonary aspergillosis. In individuals with central venous catheters in situ and profound neutropenia, invasive fungal infections are a considerable cause of morbidity and mortality. A systematic review and meta-analysis of 31 trials of antifungal prophylaxis in cancer patients after chemotherapy or haematopoietic stem-cell transplantation (HSCT), showed that antifungal prophylaxis significantly decreases all-cause mortality (RR: 0.84, 95% CI: 0.84–0.95) and the effect estimates were greater in studies with more rigorous methodology [25]. Antifungal prophylaxis was also found to be of benefit in the secondary outcomes including risk of fungal-related death (RR: 0.55, 95% CI: 0.41–0.

Alternatively, the yet-unknown compound(s) in the soil might have been converted to

tryptophan or anthranilate by the bacterial activity. Because the soil extracts did not induce the andA promoter (Nishiyama et al., 2010), such compounds might be resistant to extraction and/or poorly soluble, like lignin or humic substances. It is unlikely that levels of these inducers would be very high, and this could explain the relatively low levels of expression in soil compared with those in the in vitro cultures (Fig. 3). The andA-mutant cells failed to proliferate at the initial stage of incubation in the soil (see Fig. 5, days 7 to day 15). These data clearly indicated that the anthranilate dioxygenase genes play pivotal roles Cyclopamine cell line in the proliferation of ATCC 17616 cells in the soil. It is puzzling that loss of anthranilate dioxygenase alone abolished the growth in soil. When wild-type and andA-mutant cells were grown in the M9 minimal medium supplemented with succinate and soil extracts,

the turbidities of the culture reached a higher level than when grown in the M9 minimal medium supplemented with succinate alone, indicating that energy sources are present in the soil and that andA mutant can utilize them. Although further investigations are needed, the important role of the anthranilate dioxygenase might be to remove anthranilate, which might accumulate and have adverse effects on cells growing in the soil. As anthranilate is known to be a precursor of the signal molecules mediating intercellular see more communication for some Gram-negative bacteria (Bredenbruch et al., 2005; Farrow & Pesci, 2007; Oglesby et al., 2008), the accumulation of anthranilate might confuse the quorum sensing. We thank Dr Paul B.

Rainey for providing the IVET plasmid pUIC3. We thank Dr T. Ohmori for providing the soil sample collected from the Ehime Agricultural Experiment Station. This work was supported by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan. “
“The model organism Streptomyces coelicolor A3(2) harbors a 356-kb linear plasmid, SCP1. We report here development of a recombinational cloning method for Niclosamide deleting large segment from one telomere of SCP1 followed by replacing with the telomere of pSLA2 and sequentially inserting with the overlapping cosmids in vivo. The procedure depends on homologous recombination coupled with cleavage at telomere termini by telomere terminal protein. Using this procedure, we cloned the 81-kb avermectin and the 76-kb spinosad biosynthetic gene clusters into SCP1. Heterologous expression of avermectin production in S. coelicolor was detected. These results demonstrate the utility of SCP1 for cloning large DNA segments such as antibiotic biosynthetic gene clusters.

Alternatively, the yet-unknown compound(s) in the soil might have been converted to

tryptophan or anthranilate by the bacterial activity. Because the soil extracts did not induce the andA promoter (Nishiyama et al., 2010), such compounds might be resistant to extraction and/or poorly soluble, like lignin or humic substances. It is unlikely that levels of these inducers would be very high, and this could explain the relatively low levels of expression in soil compared with those in the in vitro cultures (Fig. 3). The andA-mutant cells failed to proliferate at the initial stage of incubation in the soil (see Fig. 5, days 7 to day 15). These data clearly indicated that the anthranilate dioxygenase genes play pivotal roles selleck screening library in the proliferation of ATCC 17616 cells in the soil. It is puzzling that loss of anthranilate dioxygenase alone abolished the growth in soil. When wild-type and andA-mutant cells were grown in the M9 minimal medium supplemented with succinate and soil extracts,

the turbidities of the culture reached a higher level than when grown in the M9 minimal medium supplemented with succinate alone, indicating that energy sources are present in the soil and that andA mutant can utilize them. Although further investigations are needed, the important role of the anthranilate dioxygenase might be to remove anthranilate, which might accumulate and have adverse effects on cells growing in the soil. As anthranilate is known to be a precursor of the signal molecules mediating intercellular 5-FU ic50 communication for some Gram-negative bacteria (Bredenbruch et al., 2005; Farrow & Pesci, 2007; Oglesby et al., 2008), the accumulation of anthranilate might confuse the quorum sensing. We thank Dr Paul B.

Rainey for providing the IVET plasmid pUIC3. We thank Dr T. Ohmori for providing the soil sample collected from the Ehime Agricultural Experiment Station. This work was supported by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan. “
“The model organism Streptomyces coelicolor A3(2) harbors a 356-kb linear plasmid, SCP1. We report here development of a recombinational cloning method for Glycogen branching enzyme deleting large segment from one telomere of SCP1 followed by replacing with the telomere of pSLA2 and sequentially inserting with the overlapping cosmids in vivo. The procedure depends on homologous recombination coupled with cleavage at telomere termini by telomere terminal protein. Using this procedure, we cloned the 81-kb avermectin and the 76-kb spinosad biosynthetic gene clusters into SCP1. Heterologous expression of avermectin production in S. coelicolor was detected. These results demonstrate the utility of SCP1 for cloning large DNA segments such as antibiotic biosynthetic gene clusters.

13 Travelers with insulin-dependent diabetes (IDD) were defined as patients with diabetes mellitus requiring daily insulin treatment, with or without additional oral anti-diabetics. Travelers with non-insulin-dependent diabetes (NIDD) were defined as patients with diabetes mellitus requiring only oral anti-diabetics. A standard questionnaire was used to collect data on socio-demographics

learn more and medical history. Items asked for were: sex, age, country of birth, history of diabetes, an immune-disorder, or another medical diagnosis, and use of medication. Participants were asked to fill out a structured diary from the day they visited the travel clinic (up to 4 weeks before departure), until 2 weeks after return from travel. Recorded in the diary were travel itinerary; any episodes of fever, diarrhea, vomiting, rhinitis, cough, and signs of skin infection; consultation with a doctor; and use of antibiotics or other medication. Fever was defined as a self-measured body temperature of 38.5°C or more. Diarrhea was defined

as loose or watery stools. Rhinitis was defined as nasal discharge or congestion. Cough could Bioactive Compound Library be dry or productive. Signs of skin infection included redness or (itching) rash, swelling, tenderness, and/or pus-like drainage. An episode of a symptomatic infection was defined as an aforementioned symptom at one or more consecutive days. The study design was not able to differentiate between non-infectious and infectious

causes. Data were collected before departure to gain information about baseline symptoms, and for 2 weeks after return to encompass incubation periods of the most (acute) travel-related infectious diseases. In the Results section, the term “travel-related” refers to the period of travel itself and the 2 weeks thereafter. The diary also provided for recording non-infectious GNAT2 symptoms and signs, such as signs of metabolic dysregulation. However, regular testing of blood glucose levels was not part of the study protocol, and hypoglycemia and hyperglycemia were not defined. Both the questionnaire and the structured diary were specifically developed for this study. According to the Dutch national guidelines on travel advice, only the travelers with medication-dependent diabetes were prescribed ciprofloxacin (500 mg 2 times a day for 3 days), to be used as immediate self-treatment after the first passage of loose or watery stools.7 Controls were advised to see a doctor in case of diarrhea with fever, blood in stools, or diarrhea persisting for 3 days or more.7 Power-analysis showed that 70 pairs were needed to prove a diarrhea outcome ratio of 2 or more, with α = 0.05 and power = 80%. This study was approved by a medical ethics committee. All participants gave their informed consent. For non-independent, non-matched characteristics, McNemar’s statistic testing was performed (spss for Windows release 15.0, SPSS Inc., Chicago, IL, USA). A p-value <0.

Two thousand and forty patients newly diagnosed with HIV/AIDS from 10 provinces in China were selected

during 2009 to 2010. Serum samples obtained from each individual were screened for HBV and HCV serum markers [HBV surface antigen (HBsAg), HBV surface antibody (HBsAb), HBV envelope antigen (HBeAg), HBV envelope antibody (HBeAb), HBV core antibody (HBcAb) and HCV antibody (HCVAb)]; liver function tests were also performed. Demographics and medical histories were collected. Of the 2040 patients, 741 (36.3%) were positive for at least one HBV and HCV serum marker; 300 (14.71%) were HCVAb positive, and 248 (12.16%) were isolated HCVAb positive; 222 (10.9%) were positive for HBsAg; 19 (0.93%) were positive for both HBsAg and HCVAb. The highest prevalence Lumacaftor of HBsAg positivity was found in Guangxi (15.31%), followed by Guangdong (15.19%) and Shanghai (14.36%). The highest prevalence of HCVAb positivity was found in Xinjiang (43.18%), followed by Henan (39.06%) and Yunnan (27.36%). The proportion of patients with abnormal liver function in patients positive for HCVAb and/or HBsAg was significantly higher than that in those who

were negative for both HCVAb and HBsAg (P < 0.001). The seroprevalence of HBV and HCV among patients newly diagnosed with HIV/AIDS in China is high. HBsAg and HCVAb positivity prevalences were found to vary significantly in different provinces in China. Patients newly diagnosed Selleck Vorinostat with HIV/AIDS and coinfected with HBV and HCV are at higher risk of abnormal liver function. It is necessary to routinely screen for HBV and HCV infection among patients newly diagnosed with HIV/AIDS.

“
“The yield of screening for acute HIV infection among general medical patients in resource-scarce settings remains unclear. Our objective was to evaluate the strategy of using pooled HIV plasma RNA to diagnose acute HIV infection in patients with negative GNA12 or discordant rapid HIV antibody tests in Durban, South Africa. We prospectively enrolled patients with negative or discordant rapid HIV antibody tests from a routine HIV screening programme in an out-patient department in Durban with an HIV prevalence of 48%. Study participants underwent venipuncture for pooled qualitative HIV RNA, and, if this was positive, quantitative RNA, enzyme immunoassay and Western blot (WB). Patients with negative or indeterminate WB and positive quantitative HIV RNA were considered acutely infected. Those with chronic infection (positive RNA and WB) despite negative or discordant rapid HIV tests were considered to have had false negative rapid antibody tests. Nine hundred and ninety-four participants were enrolled with either negative (n=976) or discordant (n=18) rapid test results. Eleven [1.1%; 95% confidence interval (CI) 0.6–2.0%] had acute HIV infection, and an additional 20 (2.0%; 95% CI 1.3–3.1%) had chronic HIV infection (false negative rapid test).

The activity was measured by decreasing the A340 nm based on the transformation of vanillin to vanillic acid. Influence on latent absorbance by the production of NAD(P)H was adjusted by molar extinction coefficients. Calculations were based on molar extinction coefficients

of vanillin (10.6 × 103; pH 7) and NAD(P)H (6.30 × 103) at 340 nm. The reaction mixture comprised 100 mM KPB, 5 mM NAD(P)+, and 0.25 mM vanillin at the final concentration. The other method investigated the enzyme’s substrate specificity and the effect of pH or temperature on it. ABT-888 Enzyme activity was determined by quantification of corresponding oxidized products from aromatic aldehydes by HPLC as described above. The reaction mixture (1.0 mL) comprised 100 mM appropriate buffer, 5 mM NAD(P)+, 2.5 mM substrates, and 0.23 and

0.26 U of purified VDH from Micrococcus sp. TA1 and B. cepacia TM1, respectively. The reaction mixture was incubated for 20 min at an appropriate temperature, after which the reaction was stopped by adding 0.2 mL of GSK458 price 0.5 N HCl. The acid produced was quantified by HPLC. One unit of enzyme activity was defined as the amount of enzyme that catalyzes the transformation of 1 μmol of vanillin to vanillic acid. The enzyme from Micrococcus sp. TA1 that catalyzes the dehydrogenation of vanillin was purified from cells as follows: after cultivation on 2 g L−1 vanillin as a carbon source with each liquid medium as described in Materials and methods, cells were harvested by centrifugation (10 000 g, 10 min, 4 °C). The pellet was resuspended in 50 mM KPB (pH 7.5) and the suspension was treated many with an ultrasonicator (201 M sonicator; Kubota, Tokyo, Japan) at 9 KHz and 170 W for 15 min; the utmost care was taken to maintain the temperature below 5 °C. The lysate was then centrifuged at 16 000 g for 20 min. The resulting supernatant was used as the cell extract. The cell extract dialyzed against 10 mM Tris-HCl buffer (pH 8.5) was applied to a 100 mL DEAE-Sepharose FF column (GE Healthcare Bio-Science, Buckinghamshire, UK). The enzyme was eluted with a

linear gradient of increasing NaCl concentration (0–0.5 M, total volume 1.5 L). The fractions with VDH activity were pooled. NaCl (2 M) was added with gentle stirring to the pooled fraction, and this solution was applied to a 30-mL butyl-Sepharose column (GE Healthcare Bio-Science). The enzyme was eluted using a gradient of decreasing NaCl concentration (2–0 M, total volume 600 mL). The active fractions were dialyzed against 10 mM Tris-HCl buffer (pH 8.5). The dialyzed enzyme solution was applied to a 1-mL Resource Q column (GE Healthcare Bio-Science). The enzyme was eluted with a linear gradient of increasing NaCl concentration (0–0.5 M, total volume 100 mL). The enzyme from B. cepacia TM1 was purified from the cell extract as follows: the cell extract of B.

In general, inhibition of fatty acid biosynthesis by the addition of cerulenin to the medium caused an increase in the polyhydroxyalkanoates and glycogen content in cells. The mutants affected in triacylglycerol accumulation used in this study also produced increased amounts of glycogen and eventually of polyhydroxyalkanoates during cultivation on gluconate in comparison

with the wild type. This effect was more evident in the mutant PDM41 than in the atf1ΩKm mutant. When the biosynthesis of triacylglycerols is impaired by inhibition of the de novo fatty acid biosynthesis Ipilimumab mw pathway or the disruption of a gene involved in triacylglycerol accumulation, carbon distribution through metabolism changes and intermediates become more available for the synthesis of glycogen and polyhydroxyalkanoates in cells. These approaches contribute toward a better understanding of storage compound metabolism and the interaction of pathways in Rhodococcus species, which could be of interest for planning further metabolic manipulation of cells for biotechnological purposes. We are grateful to Dr Alexander Steinbüchel for providing R. opacus mutant PDM41.

This study was financially supported by the Agencia Nacional de Promoción Científica y Tecnológica, Argentina (Project PME no. 216) and SCyT of the University of Patagonia San Juan Bosco. H.M. Alvarez is a career investigator and M.A. Hernández a scholarship holder of the Consejo Nacional de Investigaciones Trichostatin A cell line Científicas y Técnicas (CONICET), Argentina. “
“Antisense oligonucleotides (AS-ODN) target genes in a sequence-specific manner inhibit gene function

and have potential use as antimicrobial agents. Cell barriers, such as peptidoglycan, cell surface proteins and lipopolysaccharide membranes, prevent delivery of AS-ODN into the bacterial cell, limiting their use as an effective treatment option. The β-lactam antibiotic penicillin was examined for its ability to deliver phosphorothioate oligodeoxyribonucleotides (PS-ODNs) and γ32 P-ODN into Streptococcus mutans OMZ175. Treatment of lag-phase S. mutans OMZ175 cells with penicillin and FBA (PS-ODN targeting the fructose-biphosphate aldolase gene), resulted in prolonged suppression of growth (> 24 h) and fba expression (656.9 ± 194.4-fold O-methylated flavonoid decrease at 5 h). Suppression of both cell growth and fba expression corresponded with a greater amount of γ32 P-ODN becoming cell associated, with a maximum γ32 P-ODN concentration per cell achieved 5 h after penicillin treatment (6.50 ± 1.39 × 108 molecules per CFU). This study confirms that for S. mutans OMZ175, the peptidoglycan layer acts as a major barrier preventing AS-ODN penetration and suggests that the use of agents such as penicillin that interfere with peptidoglycan integrity can significantly increase the uptake of PS-ODN by these cells.

5; CaCl2, 2; GSK458 nmr MgSO4, 1; NaH2PO4, 1.25; NaHCO3 26; and glucose, 20; bubbled with 95% O2 and 5% CO2. Bicuculline (10 μm) or picrotoxin (100 μm) was always added to block inhibitory synaptic transmission. The signals of membrane currents were filtered at 3 kHz and digitized at 20 kHz for recording evoked climbing fiber-mediated excitatory postsynaptic currents (CF-EPSCs) or at 10 kHz for recording postsynaptic AMPA receptor-mediated currents. On-line

data acquisition and off-line data analysis were performed using PULSE software (HEKA, Lambrecht/Pfalz, Germany). Climbing fibers were stimulated via the stimulation pipette placed in the granule cell layer. Stimuli (duration, 0.1 ms; amplitude, 0–90 V) were applied at 0.2 Hz. In the experiment for the I–V relationships of the postsynaptic AMPA receptor-mediated currents, spermine (100 μm) was added to the intracellular solution and cyclothiazide (100 μm) and tetrodotoxin (0.5 μm) were added to the external solution. All experiments were carried out at 31°C. To investigate the roles of TARP γ-2 and γ-7 in synaptic expression and function

of cerebellar AMPA receptors, we generated mice deficient in γ-2 or γ-7 on the C57BL/6N background (Fig. 1A–E). A previous study reported that, when backcrossed to the C57BL/6J background, mice carrying GDC-0449 mouse the stg mutation died before weaning (Letts et al., 2003). However, our γ-2-KO mice were viable after weaning and exhibited essentially the same phenotype as the original stg mouse, including ataxic gait and head-lifting behavior. In addition, γ-2-KO mice were small in size with 73% of the body weight of their WT littermates at 8–10 weeks of age, similarly Phosphatidylethanolamine N-methyltransferase to original stg mice. On the other hand, γ-7-KO mice were viable, fertile and indistinguishable from their WT littermates. Then we crossed the two mouse lines to obtain γ-2/γ-7 double-KO (DKO) mice, which had approximately 70% of the body weight of their WT littermates. DKO mice showed much more severe ataxia than γ-2-KO mice did, as they could not walk straight and displayed frequent tumbling

and rolling as appreciated from footprint patterns (Fig. 1F). The distribution of γ-2 and γ-7 at the protein level was examined in the cerebellar cortex by producing specific antibodies. The specificity was verified by the lack of immunoreacted bands in the corresponding KO cerebella (Fig. 1E). We further noted that cerebellar content of γ-7 was reduced in γ-2-KO cerebellum, while that of γ-2 was not altered in γ-7-KO cerebellum (Fig. 1E). By immunohistochemistry, γ-2 and γ-7 were distributed at the highest levels in the cerebellum (Fig. 2A and E), the specificity of which was verified by blank immunostaining in the corresponding KO brains (Fig. 2B and F). Within the cerebellum, γ-2 was detected as clustered staining in the granular layer (i.e., synaptic glomeruli) and as punctate staining in the molecular layer (Fig. 2C and D).

By contrast, in the SCZ of wild-type (WT) mice, only a few immature (but no mature) newly generated neurons were observed, suggesting that virtually all postnatally

generated immature neurons in the SCZ were eliminated by Bax-dependent PCD. Treatment of 2-month-old WT mice with a caspase inhibitor, or with the neurotrophic factor Quizartinib cost brain-derived neurotrophic factor, promoted the survival of adult-generated neurons, suggesting that it is the absence of sufficient neurotrophic signaling in WT SCZ that triggers the Bax-dependent, apoptotic PCD of newly generated SCZ neurons. Furthermore, following focal traumatic brain injury to the posterior brain, SCZ neurogenesis in WT mice was increased, and a subset of these newly generated neurons migrated toward the injury site. These data indicate that the adult SCZ maintains a neurogenic potential that could contribute to recovery in the brain in response to the injury-induced upregulation of neurotrophic signaling. “
“The subcortical projections to the marmoset frontal pole were mapped with the use of fluorescent tracer injections. The main thalamic PLX-4720 cell line projections, which originated in both the magnocellular and parvocellular subdivisions of the mediodorsal

nucleus, were topographically organized. Our results suggest the existence of a third, caudal subdivision of this nucleus, which is likely to be homologous to the macaque’s pars densocellularis. A substantial, but not topographically organized, projection to Brodmann’s area 10 originated in the medial part of the ventral anterior nucleus. Minor thalamic projections originated in the medial pulvinar nucleus and in the midline/intralaminar nuclei. Finally, the posterior thalamic group (including the limitans and suprageniculate nuclei) sent a small projection to rostral

area 10 that has not previously been documented in primates. The main extrathalamic projections stemmed from the claustrum, which contained as many as 50% of all subcortical labelled not neurons. Minor connections originated in the hypothalamus (mainly in the lateral anterior and lateral tuberal regions), dorsal periaqueductal grey matter, basal forebrain (nucleus basalis of Meynert and horizontal limb of the diagonal band of Broca), and amygdala (basal, accessory basal and lateral nuclei). The present results, combined with recent data on the cortical projections to area 10, reveal the frontal pole as a region that integrates information from multiple neural processing systems, including high-level sensory, limbic and working memory-related structures. Although the pattern of subcortical projections is similar to that previously described in the macaque, suggesting a homologous organization, the present data also suggest functional distinctions between medial and lateral sectors of area 10.

However, increasing antibiotic resistance is threatening to undermine the effective treatment of shigellosis. Berberine is a natural isoquinoline alkaloid found in medicinal herbs, such as Rhizoma coptidis (Huanglian). Berberine selleck chemical has demonstrated a number of biological activities, including antisecretory, anti-inflammatory, antibacterial, antimalarial, antitumor and anticholesterol activities, and is widely used in the treatment of bacterial diarrhea and intestinal parasite infections in many countries (Yamamoto

et al., 1993; Iwasa et al., 1998). It has been shown that berberine has the properties of A–T base-specific DNA partial intercalation and generating singlet oxygen as a functional photosensitizer (Pilch et al., 1997; Brezova et al., 2004). Accumulated evidence suggests several mechanisms that may explain the antimicrobial activity of berberine. It has been reported that berberine interferes with the adherence of streptococci (Sun et al., 1988). It has also been demonstrated that berberine directly inhibits some Vibrio cholerae and Escherichia coli enterotoxins (Sack & Froehlich, 1982). In Leishmania donovani,

berberine exhibited an inhibitory action on macromolecular biosynthesis (Ghosh et al., 1985). Although berberine has been used in the treatment of gastrointestinal disorders, especially shigelleosis, for some time in find more China (Chang, 1959), its effect on the causative agent of shigellosis is not yet well understood. Transcript profiling based on microarray technology enables us to investigate the response of the bacterial genome to antimicrobial agents, which provides useful clues to the mechanism of

action of the agents (Fu et al., 2007). In this study, whole-genome DNA microarray was used to examine transcriptional responses elicited by berberine in Shigella flexneri. Shigella flexneri 2a strain 301 (Sf301), our sequenced strain, was used in this study (Jin et al., 2002). The bacterium was grown at 37 °C with shaking (200 r.p.m.) on cation-adjusted Mueller–Hinton broth (caMHB), a medium recommended by the Clinical and Laboratory Standards Institute (CLSI) for susceptibility testing. Berberine chloride (BC) purchased from Sigma-Aldrich Liothyronine Sodium was resolved in dimethyl sulfoxide (DMSO) and diluted with caMHB. The minimal inhibitory concentration (MIC) of BC for Sf301 was determined according to the CLSI broth macrodilution methods for bacteria that grow aerobically (Clinical and Laboratory Standards Institute, 2006). Sf301, taken from a 24-h culture in caMHB, was inoculated in the same medium until reaching an OD600 nm of about 0.05. The cultures were then allowed to continue growing at 37 °C with shaking. When they were grown to early exponential growth phase (an OD600 nm of about 0.3), BC was added from the 400 × stock dissolved in DMSO into the cultures to give final concentrations of 160, 320, and 640 μg mL−1. A control with only DMSO was also included. The final DMSO concentration for all conditions was 1% v/v.