Patients with a progressive course had a significantly (P = 0.017; OR = 18, 95% CI: 1.7–19.1) higher rate of brainstem atrophy than those with a non-progressive

http://www.selleckchem.com/products/Everolimus(RAD001).html course (monophasic or polyphasic). Presence of brainstem atrophy in the initial MRIs may predict a progressive course in patients with neuro-Behçet’s disease. “
“Background:  Family physicians measure serum levels of anti-streptolysin O antibodies (ASO) in the routine evaluation of patients with rheumatic conditions. Aim:  To evaluate the significance of elevated serum ASO titer in hospitalized patients with various clinical conditions. Patients and methods:  We retrieved the names of all patients in whom ASO serum titer was tested in our hospital during two successive years. We chose only those with titers of 1 : 160

or greater (cut-off level < 1 : 80) or with no titer. Their charts were reviewed and the causes for their hospitalization and the reasons buy MG-132 for requesting the tests were identified. We also measured the ASO serum titer in 60 healthy individuals. Results:  Six hundred and twenty-five patients were tested for ASO serum levels; 129 patients were negative. In 291 (44%) patients tests were positive at low titers (< 1 : 160). In 205 (33%) the serum titers of ASO were ≥ 1 : 160. We analyzed two groups: those with high ASO titers (≥ 1 : 160) (group 1) and those who were negative for this test (group 2). In group 1, streptococcal cultures were positive only in 14% of the patients with elevated ASO. There was no correlation between ASO serum levels and erythrocyte sedimentation rate, C-reactive protein or Amino acid rheumatoid factor. In only five individuals (8%) of the healthy cohort, was ASO significantly elevated. Conclusions: 

Elevated ASO titers can be found in various clinical conditions other than the typical post-streptococcal associated diseases. In these cases it is not necessarily accompanied by positive culture and does not correlate with inflammatory parameters. “
“The purpose of this study was to determine the prevalence of musculoskeletal complaints and rheumatic diseases in southeast of Iran. Subjects were selected based on a cluster sampling from 20 districts of urban areas in Zahedan, Iran. Subjects 15 years old and over were randomly selected and interviewed by trained interviewers in their houses. The Community Oriented Program for the Control of Rheumatic Disease (COPCORD) and Core Questionnaire (CCQ) were used in this study. The people with musculoskeletal complaints (pain, stiffness and swelling) were examined by the rheumatologist. Laboratory tests and radiographic exams were carried out when necessary to further categorize diagnoses. Data were collected from October 10, 2008 to September 15, 2009. Two thousand and one hundred subjects including 921 (43.9%) males and 1179 (56.1%) females were interviewed.

The primary outcome measure was HAART adherence during the previous month. Logistic regressions were performed to calculate the effect of each factor on adherence. All participants had HIV/AIDS and were on HAART at enrolment. Eight per cent of participants were female,

57% were African-American MK-8669 ic50 and 16% were Hispanic. Mean age was 58.1 years. Sixty-eight per cent were adherent to HAART during the last month. On univariate analysis, a preference for wanting choices, correct knowledge of recent HIV viral load level, and intention to adhere to HIV treatment were significantly associated with adherence. On multivariate analysis, only intention to adhere to HIV treatment remained statistically significant after adjusting for other factors (odds ratio 2.2; 95% confidence interval 1.1 to 4.3). Intention to adhere to HIV treatment was significantly associated with self-reported adherence to HAART. Interventions that bolster patients’ intentions to adhere to HIV treatment during clinical encounters may

improve adherence to HAART and HIV control. “
“The renal elimination of tenofovir (TFV) may be subject to renal drug−drug interactions that may increase the risk of kidney injury. Case reports indicated that diclofenac might increase TFV-associated nephrotoxicity via a drug−drug Selleckchem PD98059 interaction, leading to an increased intracellular TFV concentration in proximal tubular cells. A retrospective analysis of data for all patients from the Frankfurt HIV Cohort (FHC) who had diclofenac prescriptions between January 2008 and June 2012 was carried out. Among 89 patients with diclofenac use, 61 patients (68.5%) were treated with tenofovir disoproxil fumarate (TDF) and 28 patients (31.5%) were treated with TDF-sparing combination antiretroviral (-)-p-Bromotetramisole Oxalate therapy (cART). Thirteen patients (14.6%) developed acute kidney injury (AKI) shortly after initiating diclofenac treatment. AKI occurred exclusively in TDF-treated patients, although all had previously stable renal function. All cases were accompanied by new onset of at least

two parameters indicating proximal tubular damage, such as normoglycaemic-glucosuria and hypophosphataemia. TFV-associated nephrotoxicity was demonstrated by renal biopsy in four cases. Additionally, 11.5% of patients on TDF treatment developed new-onset proximal tubular damage, while having a preserved glomerular filtration rate. In contrast, diclofenac did not affect renal function in patients with TDF-sparing cART, as only one case of isolated hypophataemia was observed in these patients. In univariate analysis, risk factors for AKI were TDF-containing cART (P = 0.0076) and pre-existing hypophosphataemia (P = 0.0086). Drug−drug interaction caused by diclofenac could exacerbate TFV-associated nephrotoxicity. Diclofenac should be used with caution in patients on TDF therapy, especially in those with hypophosphataemia.

PCC 7120, it has indeed GSI-IX ic50 been shown that the amount of DNA in the two newborn daughter cells after cell division is not always identical, but can vary (Hu et al., 2007; Schneider et al., 2007). An additional advantage is gene redundancy,

which opens the possibility that under unfavorable conditions, mutations are induced in some genome copies, whereas the wildtype information is retained in others. It has indeed been shown that heterozygous cells of S. elongatus PCC 7942 and of Synechocystis PCC 6803 can be selected, at least under laboratory conditions (Labarre et al., 1989; Spence et al., 2004; Takahama et al., 2004; Nodop et al., 2008). Heterozygous strains have also been selected of two halophilic and methanogenic archaea, Haloferax volcanii and Methanococcus maripaludis. In both cases, it was shown that in the absence of selection gene conversion leads to the equalization of genomes and reappearance of homozygous cells (Hildenbrand et al., 2011; Lange et al., 2011). By analogy, we predict that gene conversion also operates in oligo- and

polyploid species of cyanobacteria. The higher efficiency of gene replacement with linear DNA compared with circular DNA in Synechocystis PCC 6803 indicates that this is really the case (Labarre et al., 1989). This work was supported by grant So264/16-1 of the German Research Council (Deutsche Forschungsgemeinschaft). We thank Annegret Wilde (University of Giessen, Germany) for the motile and the GT Synechocystis PCC 6803 strains, Wolfgang R. Hess for S. elongatus Selleckchem Regorafenib PCC 7942 and Synechococcus sp. WH7803, and both for very valuable advice concerning growth of cyanobacteria. We thank Enrico Schleiff for the possibility to grow cyanobacterial cultures in his light incubator. We are grateful to two reviewers who were patient with us as non-experts of cyanobacteria,

and gave us very good suggestions and literature references. “
“1-Aminocyclopropane-1-carboxylate (ACC) deaminase is commonly produced by plant growth-promoting rhizobacteria (PGPR) and has been suggested to facilitate the growth and stress tolerance of hosts via a reduction in levels of ethylene. However, the regulatory mechanism of ACC deaminase (AcdS) protein within host plant cells is largely unknown. Here, we demonstrated beneficial effects and post-translational modification of PGPR-originated AcdS triclocarban proteins in plants. Compared with the wild-type, transgenic Arabidopsis expressing the Pseudomonas fluorescens acdS (PfacdS) gene displayed increased root elongation and reduced sensitivity to 10 μM exogenous ACC, an ethylene precursor. Arabidopsis expressing PfacdS also showed increased tolerance to high salinity (150 mM NaCl). PfAcdS proteins accumulated in transgenic Arabidopsis were rapidly degraded, which was potentially mediated by the 26S proteasome pathway. The degradation of PfAcdS was alleviated in the presence of exogenous ACC.

7,8 Estimations of TD risk in Bangkok have traditionally been extrapolated from find more post-Thailand travel surveys and descriptions of contaminated food sources in Bangkok.6,9–11 Although food bacterial prevalence studies provide important information, contaminated retail food sources do not often translate to TD risk. Bacterial prevalence results from raw retail food samples in Bangkok are similar to equivalent studies performed in places considered “low risk” for TD such as Washington,

DC, Alberta, Canada, and Ireland.11–14 Inadequate public health practices is a more important risk than initial contamination of specific food and beverage items, and countries demonstrating decreased diarrhea rates over time have done so by the improvement of infrastructure and public health practices.15–18 Estimates place the population of Bangkok at 10 to 15 million people. Bangkok is an international business destination and rivals Hong Kong and Singapore as a modern Asian city. Bangkok

restaurants have not been examined as a source of bacterial pathogens, yet eating at restaurants, in multiple TD epidemiologic studies, is statistically associated with TD.17–19 This study seeks to ascertain a traveler’s risk of exposure to bacterial gastric pathogens, a rough measure of diarrhea acquisition risk, through the www.selleckchem.com/products/PD-0332991.html sampling of restaurant meals at the end point in the food preparation and serving process (ie, the customer’s plate) and to assess antibiotic resistance patterns of the identified pathogens. The most prevalent bacterial pathogens known to cause disease in the limited studies of TD etiology in Thailand, namely Campylobacter spp. and Salmonella spp., were assessed

along with Arcobacter, an emerging Campylobacter-like organism.6,20–23 Current guidance on antibiotic treatment for TD in Thailand focuses on azithromycin because of the high degree of ciprofloxacin resistant Campylobacter spp. However, azithromycin resistant Campylobacter spp. is present selleck inhibitor in Thailand and antibiotic resistance patterns should be further monitored.24,25 A cross-sectional tourist restaurant survey was conducted in March of 2009. A total of 121 Bangkok restaurants were identified from the top two selling Bangkok guide books on Amazon.com. A random sample of 35 restaurants were obtained and sampled. These 35 restaurants represent a good cross-section of downtown Bangkok visited by tourists (Figure 1). Restaurants were visited by study staff as customers over a 3-week time period in March 2009, which is during the dry season and a high tourist period. Two meals were ordered for sampling at each restaurant; one from the recommended meal in the guide book and another from a meal likely to contain a bacterial pathogen (raw meats, fresh salads, etc.).

Participants were given an example of think-aloud interview technique and then asked to verbalize their thoughts

as they answered each question in the questionnaire and to indicate the reasons for providing the answers. Prompts (calendars, maps, and festival dates) were provided and on completion of the interview all participants were administered 24 structured follow-up probe questions. Use of prompts was observed and recorded. Scripted probes were used; responses were recorded by the investigator and subsequently analyzed. Items from the cognitive interviews were refined and incorporated into the final version of the questionnaire. We were not able to find copies (printed or electronic) of any questionnaires used in published travel-related Roscovitine datasheet studies, and none of the travel studies reported a process of validation. Thirty-four pooled items were selected for inclusion in the pre- and post-travel questionnaires (version 2). Sixty-four travelers were recruited to the prospective cohort study and completed the pre-travel questionnaire; the pilot study included 23 who had returned to complete the post-travel questionnaires. The remaining 38 travelers had not returned from travel and 3 were lost to follow-up. Age of the participants

ranged from 16 to 71 (median: 36) years, 42% were male, and 27% were overseas born. Most (62.5%) were tourists. Item-specific and general problems were identified by steps 3 and 4. Item-specific FG-4592 problems were mainly related to suboptimal clarity and an inadequate number of response categories provided. Table 1 provides examples of the item-specific problems identified, classification within the QAS framework, and the final revised many items. In addition, feedback by travelers, together with observed and self-reported difficulties in the pilot study, resulted in an expansion of the draft questionnaire items from 34 to 39. Seven of 19 post-travel

questionnaire items and 7 of 15 pre-travel questionnaire items were revised. Participants’ difficulties included deciding which destinations were “rural” locations and selection of appropriate traveler type category: definitions were therefore provided in the questionnaires. Some problems applied to multiple items across the questionnaire relating to QAS-99 categories of knowledge and memory. It was recognized that complicated travel itineraries and longer travel durations would be difficult to recall and record despite follow-up consultation within 2 weeks of return from travel. Open-ended questions were not selected for the categories of accommodation type or travel activities, as it was judged too difficult a recall task for travelers with long travel durations or complicated itineraries. Instead, a list of response options was provided. Some travelers did not report destination countries or health episodes in their correct temporal order.

, 2008). Because

IL-1β represents a major pro-inflammatory cytokine involved in the induction of miR-146a (Taganov et al., 2006; Nakasa et al., 2008; Sheedy & O’Neill, 2008), it is possible that expression of miR-146a in astrocytes may represent an attempt to modulate the inflammatory response triggered by this cytokine. Accordingly, recent studies identify miR-146a as a key regulator in a feedback system whereby induction of nuclear factor kappa-B selleck chemical (NFkB) through a myeloid differentiation factor 88 (MyD88)-dependent pathway may upregulate the miR-146a, which in turn could downregulate the levels of two key adapter molecules, IL-1RI-associated protein kinases-1 (IRAK1) and -2, and TNF receptor-associated factor 6 (TRAF6) downstream of TLR and cytokine receptors, reducing the activity of this inflammatory pathway (Taganov et al., 2006; Hou et al., 2009). Navitoclax concentration These observations are particularly interesting considering the known proconvulsant action of IL-1β mediated by the IL-1 receptor type 1,

as well as the recently reported role of TLR-signalling pathways in epilepsy (Vezzani et al., 2008; Maroso et al., 2009), and suggest that miR-146a induction could function in fine-tuning the response to cytokines in TLE during epileptogenesis. The upregulation of miR-146a observed in the chronic epileptic phase in the post-SE model of TLE was confirmed in human HS specimens of patients undergoing surgery

for pharmacologically refractory TLE. In situ hybridization analysis of miR-146a in human control hippocampus and HS specimens demonstrated expression in neuronal cells. In contrast (as observed in the post-SE rat hippocampus), the expression in glial cells was detected only in tissue of patients with HS, particularly much in regions with prominent gliosis. Expression of the miR-146a was observed in neurons and in reactive astrocytes in HS tissue. Neurons constitute an additional source of pro-inflammatory cytokines (including IL-1β), potentially contributing to the inflammatory pathology observed in TLE (Ravizza et al., 2008). Thus, the neuronal expression of miR-146a may also represent an attempt to regulate this inflammatory pathway. A physiological mechanism of defence against activation of inflammatory pathways during epileptogenesis is represented by induction of inhibitory factors, such as CFH (Boon et al., 2009), an important repressor of inflammatory signalling. This factor inhibits excessive activation of the complement cascade, which is prominently activated in both experimental and human TLE (Aronica et al., 2007). Interestingly, CFH has been identified as a target of miR-146a. For instance, in AD brains, upregulation of miR-146a has been linked to downregulation of CFH (Lukiw et al., 2008).

, 2008). Because

IL-1β represents a major pro-inflammatory cytokine involved in the induction of miR-146a (Taganov et al., 2006; Nakasa et al., 2008; Sheedy & O’Neill, 2008), it is possible that expression of miR-146a in astrocytes may represent an attempt to modulate the inflammatory response triggered by this cytokine. Accordingly, recent studies identify miR-146a as a key regulator in a feedback system whereby induction of nuclear factor kappa-B MLN8237 (NFkB) through a myeloid differentiation factor 88 (MyD88)-dependent pathway may upregulate the miR-146a, which in turn could downregulate the levels of two key adapter molecules, IL-1RI-associated protein kinases-1 (IRAK1) and -2, and TNF receptor-associated factor 6 (TRAF6) downstream of TLR and cytokine receptors, reducing the activity of this inflammatory pathway (Taganov et al., 2006; Hou et al., 2009). OSI-906 clinical trial These observations are particularly interesting considering the known proconvulsant action of IL-1β mediated by the IL-1 receptor type 1,

as well as the recently reported role of TLR-signalling pathways in epilepsy (Vezzani et al., 2008; Maroso et al., 2009), and suggest that miR-146a induction could function in fine-tuning the response to cytokines in TLE during epileptogenesis. The upregulation of miR-146a observed in the chronic epileptic phase in the post-SE model of TLE was confirmed in human HS specimens of patients undergoing surgery

for pharmacologically refractory TLE. In situ hybridization analysis of miR-146a in human control hippocampus and HS specimens demonstrated expression in neuronal cells. In contrast (as observed in the post-SE rat hippocampus), the expression in glial cells was detected only in tissue of patients with HS, particularly Nintedanib (BIBF 1120) in regions with prominent gliosis. Expression of the miR-146a was observed in neurons and in reactive astrocytes in HS tissue. Neurons constitute an additional source of pro-inflammatory cytokines (including IL-1β), potentially contributing to the inflammatory pathology observed in TLE (Ravizza et al., 2008). Thus, the neuronal expression of miR-146a may also represent an attempt to regulate this inflammatory pathway. A physiological mechanism of defence against activation of inflammatory pathways during epileptogenesis is represented by induction of inhibitory factors, such as CFH (Boon et al., 2009), an important repressor of inflammatory signalling. This factor inhibits excessive activation of the complement cascade, which is prominently activated in both experimental and human TLE (Aronica et al., 2007). Interestingly, CFH has been identified as a target of miR-146a. For instance, in AD brains, upregulation of miR-146a has been linked to downregulation of CFH (Lukiw et al., 2008).

During period one

the patients injected the insulin bolus before the meal and, during period two, after the Estrogen antagonist meal. The variability of blood glucose (BG) was assessed by low BG indices (LBGI) and high BG indices (HBGI) – the measure of the variability of low and high BG readings. Their sum (LBGI + HBGI) gives the BG risk index (BGRI) – a measure of overall variability and deviations towards hypo- and hyperglycaemia. Six patients were on CSII and six on MDI. The number of meals, number of insulin injections and average BG were not different between the groups. LBGI and the number of hypoglycaemic events were not affected by the method of injection. BGRI were significantly higher for post-meal injection, mainly due to increased hyperglycaemia (p=0.003). The increased HBGI and BGRI were more prominent in CSII (p=0.05). These differences were found for the 72-hour variability but not when testing 2 hours post-prandially.

It was Saracatinib concluded that injecting insulin prior to the meal can reduce the overall glucose variability, and remains the preferred method of injection. Larger studies are needed in order to reinforce these results. Copyright © 2012 John Wiley & Sons. “
“Gestational diabetes mellitus (GDM) is common, with an average prevalence in England and Wales of approximately 3.5%. It is associated with a 70% lifetime risk of developing type 2 diabetes mellitus (T2DM) for the women in the long term. It is therefore important to continue lifelong monitoring for abnormalities of glucose metabolism. There is a lack of international consensus on the best postpartum screening test, its timing, and the frequency and duration of long-term follow up after GDM. In general, screening rates are suboptimal

across the globe with perhaps an optimistic trend in recent years with just over half of the women completing Cyclin-dependent kinase 3 postpartum screening. Postpartum diabetes screening may detect T2DM and enable early treatment of hyperglycaemia, reducing the risk of adverse fetal outcomes in subsequent pregnancies and maternal microvascular complications. Screening can also identify women who might benefit from diabetes prevention interventions. Metformin has been shown to reduce the rate of diabetes development following delivery by 50% and should be considered in all cases of GDM if tolerated. Copyright © 2010 John Wiley & Sons. “
“Appropriate management of diabetes during labor and delivery plays a significant role in ensuring the wellbeing of the mother and neonate. Maternal hyperglycemia is the major cause of neonatal hypoglycemia. The role of the physician during this period is to maintain maternal euglycemia in order to prevent ketoacidosis and reduce the risk of neonatal hypoglycemia. Management of diabetes during labor should follow an established protocol in a dedicated center with a neonatal care unit equipped and staffed to deliver the most sophisticated level of care.

This observation is consistent with previous work from our lab indicating that total intake, not the length of the self-administration history, is responsible for the neurochemical changes that occur following cocaine self-administration (Calipari et al., 2013). Because glucose utilization was assessed immediately after the final reinforcer in the 30-day self-administration group from Macey

et al. (2004) and rates were significantly lower than controls, it suggests that not only are the circuits depressed in the absence of cocaine, but also that find more cocaine failed to produce sufficient effects to ‘normalize’ circuits. Continued drug-taking in addicted individuals has been suggested to occur as compensatory behavior to ‘normalize’ a baseline dysregulated state (Koob & Le Moal, 1997; Koob, 2009). It is important to differentiate the effects of cocaine-induced alterations of neural networks while cocaine is present with those of cocaine self-administration on functioning in the absence of drug, as they have very different implications for the functioning of the brain at baseline. The mesocorticolimbic dopamine system mediates many of the reinforcing and rewarding effects of cocaine (Pierce & Kumaresan,

2006), and because neuroadaptations resulting from chronic drug exposure are often opposite from the acute effects, it is not surprising that there were reductions in the activity of these regions following cocaine self-administration. Previous work has demonstrated reduced http://www.selleckchem.com/products/Belinostat.html BCKDHA function of the striatal dopamine system at a similar time point following cessation of cocaine self-administration, as well as the development

of tolerance to the neurochemical effects of cocaine (Ferris et al., 2011). Furthermore, these functional impairments are present 18 h following the final cocaine self-administration session (Mateo et al., 2005; Ferris et al., 2011, 2012; Calipari et al., 2012), and persist for up to 2 weeks following cocaine exposure (Ferris et al., 2012). These reductions in dopamine function have been observed using both fast scan cyclic voltammetry and microdialysis where it was found that 18–24 h of withdrawal from cocaine self-administration resulted in reduced dopamine release and uptake, as well as reduced baseline dopamine overflow, respectively (Weiss et al., 1992; Maisonneuve et al., 1995; Ferris et al., 2011, 2012; Calipari et al., 2012, 2013; but see Hooks et al., 1994; Meil et al., 1995). The current data agree with these observations in that functional activity was significantly reduced in the terminal fields of the ventral tegmental area, namely the nucleus accumbens and caudate putamen (Koeltzow & White, 2003).

1, CU459141.1, and

CP001182.1). Random amplification of polymorphic DNA (RAPD) analysis was subsequently used to discriminate the A. baumannii strains. Primers Wil2 (Williams et al., 1993) and 1247 (Akopyanz et al., 1992) previously used for typing other bacteria were applied. Some other representatives of the genus of Acinetobacter such as A. lwoffii (six strains), A. anitratus (4), and A. calcoaceticus (3) and several other gram-negative microorganisms such as P. aeruginosa, Escherichia coli, Yersinia pseudotuberculosis, Yersinia enterocolitica, Klebsiella pneumoniae, Decitabine cost Klebsiella oxytoca, Enterobacter cloacae, Pasteurella multocida, and Salmonella Enteritidis (three strains of each species) were used in the research. All bacteria were grown in Luria–Bertani (LB) broth or nutrient agar (Himedia Laboratories Pvt. Limited, India) at 37 °C. Clinical materials and in-hospital environmental samples were used for phage isolation. Nonliquid samples were kept in 0.1 M Tris–HCl buffer, pH 7.0. The samples were cleared by low-speed centrifugation (7000 g for 30 min.) followed by filtration of the supernatants through 1.20- and 0.45-μm-pore-size membrane filters (Millipore) to remove bacterial debris. The purified filtrates were concentrated by ultracentrifugation at 85 000 g at 4 °C for 2 h (Beckman SW28 rotor). The spot test method as well as the plaque assay (Adams, 1959) was used to screen for the presence of lytic

phage activity http://www.selleckchem.com/products/ink128.html in the resultant concentrates using clinical A. baumannii strains of different RAPD groups. The plates were incubated overnight at 37 °C and examined for zones of lysis or plaques formation. Single plaque isolation was used to obtain pure phage stock. For that a single plaque formed on the A. baumannii lawn was picked

up in SM buffer (10 mM Tris–HCl, pH 7.5, 10 mM MgSO4 × 7 H2O, and 100 mM NaCl) and replated three times. Phage AP22 was propagated using liquid culture of identified A. baumannii clinical strain 1053 (OD600 nm of 0.3) at multiplicity of infection (MOI) of 0.1. The incubation was performed at 37 °C until complete lysis, only and then chloroform was added. Bacterial debris was pelleted by centrifugation at 7000 g for 30 min. The phage lysate was concentrated by ultracentrifugation at 85 000 g at 4 °C for 2 h (Beckman SW28 rotor). The resultant pellet was carefully mixed with SM buffer and centrifuged at 13 000 g. Supernatant was treated with DNase (1 μg mL−1) and RNase (1 μg mL−1) at 37 °C. The nucleases were removed with chloroform. The phage preparation with the titer of 1012–1013 PFU mL−1 was purified by cesium chloride equilibrium gradient centrifugation at 100 000 g (Beckman SW41 rotor) for 24 h (Sambrook et al., 1989). Host specificity of the phage was determined by double-layer method. Onto the surface of M9 medium (Sambrook et al., 1989) plates, 0.3 mL of liquid bacterial culture (108–109 PFU mL−1) and 4 mL of soft agar (LB broth supplemented with 0.