However, they should have high encapsulation efficiency with sustained and prolonged intracellular antibiotic release. For example, our core–shell nanostructures can incorporate up to 25% by weight of gentamicin, and about 25–30% of the gentamicin is released over 24 h in phosphate buffer saline at pH 7.4 (Ranjan et al., 2010a ,b). But, as the gentamicin begins to leave the complex, the net anionic character of the complexes increases. As this occurs, greater electrostatic attraction between the polymer and gentamicin slows or

completely prevent further release. Therefore, nanocarrier needs to be modified such that they degrade slowly to release 100% of the encapsulate drug. We recently reported

biodegradable silica xerogel nanocarrier for complete drug release (Seleem et al., 2009a ,b). Xerogel nanostructures are prepared by a sol–gel process. This involves formation A-769662 in vivo of a colloidal suspension (sol) that acts as a precursor for globally connected integrated solid matrix (gel) that can be dried to form xerogel (Quintanar-Guerrero et al., 2009). The xerogels can be fabricated and tuned at low temperatures to carry biologically active agents like gentamicin (Xue et al., 2006). Silica xerogels nanostructures prepared by our technique can incorporate 17% gentamicin by weight and releases 90% of gentamicin in 30 h in vitro. Gentamicin release from these nanostructures Mitomycin C nmr is biphasic. A total of 20–25% of drug is initially released at a burst rate followed by a slower and steady state. Biphasic release may be problematic in vivo because burst release can result in encapsulated

drug acting similar to its free form. This is reflected all in an incomplete in vivo clearance of intracellular Salmonella in the livers (1.15 log reduction in CFU) and spleen (0.41 log reduction in CFU). Therefore, although the results are encouraging, careful engineering and chemical principles are required in particle synthesis to address these issues before further clinical application. This review summarized the recent findings on targeting of intracellular pathogens especially Salmonella. As discussed, incorporation of antimicrobials in a nanocarrier provides a novel method for intracellular drug delivery and enhancing their killing effect. However, complete eradication of intracellular pathogens using this methodology is yet to be realized. Targeted drug delivery and their intracellular bioactivity are two separate issues. In our opinion, antibacterial nanomedicine in its true sense is the delivery of targeted drug to the subcellular niche where a bacterium resides. Currently available technologies deliver drugs to the cell endosome or cytoplasm and hence may not be fully targeted. Endosomal or cytoplasmic delivery exposes drug initially to the cellular microenvironment prior to their interaction with the bacteria.

However, they should have high encapsulation efficiency with sustained and prolonged intracellular antibiotic release. For example, our core–shell nanostructures can incorporate up to 25% by weight of gentamicin, and about 25–30% of the gentamicin is released over 24 h in phosphate buffer saline at pH 7.4 (Ranjan et al., 2010a ,b). But, as the gentamicin begins to leave the complex, the net anionic character of the complexes increases. As this occurs, greater electrostatic attraction between the polymer and gentamicin slows or

completely prevent further release. Therefore, nanocarrier needs to be modified such that they degrade slowly to release 100% of the encapsulate drug. We recently reported

biodegradable silica xerogel nanocarrier for complete drug release (Seleem et al., 2009a ,b). Xerogel nanostructures are prepared by a sol–gel process. This involves formation GDC-0980 concentration of a colloidal suspension (sol) that acts as a precursor for globally connected integrated solid matrix (gel) that can be dried to form xerogel (Quintanar-Guerrero et al., 2009). The xerogels can be fabricated and tuned at low temperatures to carry biologically active agents like gentamicin (Xue et al., 2006). Silica xerogels nanostructures prepared by our technique can incorporate 17% gentamicin by weight and releases 90% of gentamicin in 30 h in vitro. Gentamicin release from these nanostructures Akt inhibitor is biphasic. A total of 20–25% of drug is initially released at a burst rate followed by a slower and steady state. Biphasic release may be problematic in vivo because burst release can result in encapsulated

drug acting similar to its free form. This is reflected Ketotifen in an incomplete in vivo clearance of intracellular Salmonella in the livers (1.15 log reduction in CFU) and spleen (0.41 log reduction in CFU). Therefore, although the results are encouraging, careful engineering and chemical principles are required in particle synthesis to address these issues before further clinical application. This review summarized the recent findings on targeting of intracellular pathogens especially Salmonella. As discussed, incorporation of antimicrobials in a nanocarrier provides a novel method for intracellular drug delivery and enhancing their killing effect. However, complete eradication of intracellular pathogens using this methodology is yet to be realized. Targeted drug delivery and their intracellular bioactivity are two separate issues. In our opinion, antibacterial nanomedicine in its true sense is the delivery of targeted drug to the subcellular niche where a bacterium resides. Currently available technologies deliver drugs to the cell endosome or cytoplasm and hence may not be fully targeted. Endosomal or cytoplasmic delivery exposes drug initially to the cellular microenvironment prior to their interaction with the bacteria.

5′-Nucleotidase activity has been described in bacteria, plant cells and in various vertebrate tissues (Zimmermann, 1992). Little information is available about ecto-5′-nucleotidase and extracellular free adenosine in the pathogenic processes of fungi. In this work, we identified some biochemical properties of C. parapsilosis ecto-5′-nucleotidase that could be involved in the release of free adenosine into extracellular

medium. The detection of cell surface-located AMP hydrolysis was confirmed and 5′-nucleotidase activity in supernatant was <20% of that found in intact cells (Fig. 1). In all conditions used during the incubation periods, the cells were viable, suggesting that the low 5′-AMP hydrolysis observed in the supernatant could be attributed to secreted enzymes. A phosphatase inhibitor, sodium orthovanadate (de Almeida-Amaral et Trichostatin A al., 2006; Kiffer-Moreira et selleck chemical al., 2007a; Leite et al., 2007; Amazonas et al., 2009), inhibited ectophosphatase

on the surface of C. parapsilosis; however, no inhibitory effect was seen in the ecto-5′-nucleotidase activity (Fig. 4). The optimum pH for this nucleotidase enzyme is in the acidic range, with maximal activity at a pH of 4.5 (Fig. 3b). Interestingly, this result is different from that observed in T. vaginalis strains, in which the optimum pH was in the neutral range (Tasca et al., 2003), and in mammalian ecto-5′-nucleotidase, not in which maximal enzyme activity was obtained in the alkaline pH range of 7–8 (Zimmermann, 1992). This assay also rules out the possibility of 5′-AMP hydrolysis due to the action of ecto-ATPase because the activity of ecto-ATPase is primarily in the alkaline pH range (Kiffer-Moreira et al., 2010). Candida parapsilosis ecto-5′-nucleotidase activity is independent of divalent cations, but it can be activated by Ca2+ and Mg2+ (Fig. 3a). These same characteristics

were observed for 5′-nucleotidase activity in T. vaginalis (Tasca et al., 2003). The enzyme also showed a high sensitivity to ammonium molybdate, a classical nucleotidase inhibitor (Gottlieb & Dwyer, 1983; Borges et al., 2007), in which concentrations above 0.5 mM abolished the enzyme activity altogether (Fig. 5). Intact cells of C. parapsilosis were able to hydrolyze all substrate monophosphates, except 3′-AMP. As described in other cells, 5′-nucleotidases hydrolyze exclusively nucleoside 5′-monophosphates, showing no activity for 3′-monophosphates. 5′-AMP is commonly known as the most hydrolyzable nucleotide by 5′-nucleotidase (Zimmermann, 1992, 1996; Borges et al., 2007). Nevertheless, C. parapsilosis ecto-5′-nucleotidase activity seems to exhibit no significant difference in hydrolyzing 5′-AMP, 5′-UMP and 5′-IMP as substrates (Fig. 2).

5′-Nucleotidase activity has been described in bacteria, plant cells and in various vertebrate tissues (Zimmermann, 1992). Little information is available about ecto-5′-nucleotidase and extracellular free adenosine in the pathogenic processes of fungi. In this work, we identified some biochemical properties of C. parapsilosis ecto-5′-nucleotidase that could be involved in the release of free adenosine into extracellular

medium. The detection of cell surface-located AMP hydrolysis was confirmed and 5′-nucleotidase activity in supernatant was <20% of that found in intact cells (Fig. 1). In all conditions used during the incubation periods, the cells were viable, suggesting that the low 5′-AMP hydrolysis observed in the supernatant could be attributed to secreted enzymes. A phosphatase inhibitor, sodium orthovanadate (de Almeida-Amaral et selleck chemicals llc al., 2006; Kiffer-Moreira et Small molecule library al., 2007a; Leite et al., 2007; Amazonas et al., 2009), inhibited ectophosphatase

on the surface of C. parapsilosis; however, no inhibitory effect was seen in the ecto-5′-nucleotidase activity (Fig. 4). The optimum pH for this nucleotidase enzyme is in the acidic range, with maximal activity at a pH of 4.5 (Fig. 3b). Interestingly, this result is different from that observed in T. vaginalis strains, in which the optimum pH was in the neutral range (Tasca et al., 2003), and in mammalian ecto-5′-nucleotidase, DNA ligase in which maximal enzyme activity was obtained in the alkaline pH range of 7–8 (Zimmermann, 1992). This assay also rules out the possibility of 5′-AMP hydrolysis due to the action of ecto-ATPase because the activity of ecto-ATPase is primarily in the alkaline pH range (Kiffer-Moreira et al., 2010). Candida parapsilosis ecto-5′-nucleotidase activity is independent of divalent cations, but it can be activated by Ca2+ and Mg2+ (Fig. 3a). These same characteristics

were observed for 5′-nucleotidase activity in T. vaginalis (Tasca et al., 2003). The enzyme also showed a high sensitivity to ammonium molybdate, a classical nucleotidase inhibitor (Gottlieb & Dwyer, 1983; Borges et al., 2007), in which concentrations above 0.5 mM abolished the enzyme activity altogether (Fig. 5). Intact cells of C. parapsilosis were able to hydrolyze all substrate monophosphates, except 3′-AMP. As described in other cells, 5′-nucleotidases hydrolyze exclusively nucleoside 5′-monophosphates, showing no activity for 3′-monophosphates. 5′-AMP is commonly known as the most hydrolyzable nucleotide by 5′-nucleotidase (Zimmermann, 1992, 1996; Borges et al., 2007). Nevertheless, C. parapsilosis ecto-5′-nucleotidase activity seems to exhibit no significant difference in hydrolyzing 5′-AMP, 5′-UMP and 5′-IMP as substrates (Fig. 2).

An avidity index of < 80% is reported as indicative of recently acquired HIV infection with an estimated time of crossing this threshold of 155 days [5]. National reporting of the proportion of recently infected among newly diagnosed persons by the HPA takes into account available clinical data

in order to reduce misclassification: patients with advanced infection (a CD4 count < 200 cells/μL or an AIDS-defining illness) or on antiretroviral therapy (including pre-exposure or post-exposure prophylaxis treatment) are assigned to ‘long-standing’ regardless of the avidity index. The laboratory result, which is available at clinic level, does not take into account these additional data and it is therefore the responsibility of the clinician to include clinical and behavioural factors during discussions with patients. Details of the RITA programme Selleckchem HDAC inhibitor and caveats in interpreting the results are provided to clinics as information sheets and are made available on the HPA website [3]. Over the past decade,

tests of recent HIV infection have been applied in epidemiological studies to estimate HIV incidence in defined populations using a single aliquot. More recently, Selumetinib cell line TRI have been conducted on aliquots of newly diagnosed persons as part of routine public health monitoring in parts of the USA, France and E&NI [2, 6, 7]. While results have previously been discussed with patients in the context of partner notification in local pilots in the USA [8], E&NI are currently the only countries where results are available Methocarbamol to patients at the clinician’s discretion. We conducted a survey among HIV clinicians and health advisors to investigate the role of RITA in clinical practice and during contact tracing efforts, and to report any patient adverse events. An online questionnaire

using Surveymonkey® (Palo Alto, California) was distributed to clinicians and health advisors via the British HIV Association (BHIVA) membership email list in February 2011. Before the launch the survey was piloted among 15 HIV specialists. BHIVA members were given 1 month to respond to the survey. Questions covered processing of TRI at individual centres, interpretation of results in the clinical setting, patients’ responses during consultations and the role of RITA results when tracing sexual contacts of patients. Some exploratory questions allowed more than one answer. Survey results were collected online and analysed using Microsoft Excel. Results were stratified by centre and job title of respondents. Forty-two HIV specialists replied to the survey from 32 HIV centres (response rate 32 of 90; 36%), which provide the RITA service in E&NI. Respondents constituted 30 consultants or associate specialists (71%), nine junior doctors (21%), one nurse consultant, one health advisor and one virologist (2% each).

An avidity index of < 80% is reported as indicative of recently acquired HIV infection with an estimated time of crossing this threshold of 155 days [5]. National reporting of the proportion of recently infected among newly diagnosed persons by the HPA takes into account available clinical data

in order to reduce misclassification: patients with advanced infection (a CD4 count < 200 cells/μL or an AIDS-defining illness) or on antiretroviral therapy (including pre-exposure or post-exposure prophylaxis treatment) are assigned to ‘long-standing’ regardless of the avidity index. The laboratory result, which is available at clinic level, does not take into account these additional data and it is therefore the responsibility of the clinician to include clinical and behavioural factors during discussions with patients. Details of the RITA programme PCI-32765 and caveats in interpreting the results are provided to clinics as information sheets and are made available on the HPA website [3]. Over the past decade,

tests of recent HIV infection have been applied in epidemiological studies to estimate HIV incidence in defined populations using a single aliquot. More recently, selleck inhibitor TRI have been conducted on aliquots of newly diagnosed persons as part of routine public health monitoring in parts of the USA, France and E&NI [2, 6, 7]. While results have previously been discussed with patients in the context of partner notification in local pilots in the USA [8], E&NI are currently the only countries where results are available Rebamipide to patients at the clinician’s discretion. We conducted a survey among HIV clinicians and health advisors to investigate the role of RITA in clinical practice and during contact tracing efforts, and to report any patient adverse events. An online questionnaire

using Surveymonkey® (Palo Alto, California) was distributed to clinicians and health advisors via the British HIV Association (BHIVA) membership email list in February 2011. Before the launch the survey was piloted among 15 HIV specialists. BHIVA members were given 1 month to respond to the survey. Questions covered processing of TRI at individual centres, interpretation of results in the clinical setting, patients’ responses during consultations and the role of RITA results when tracing sexual contacts of patients. Some exploratory questions allowed more than one answer. Survey results were collected online and analysed using Microsoft Excel. Results were stratified by centre and job title of respondents. Forty-two HIV specialists replied to the survey from 32 HIV centres (response rate 32 of 90; 36%), which provide the RITA service in E&NI. Respondents constituted 30 consultants or associate specialists (71%), nine junior doctors (21%), one nurse consultant, one health advisor and one virologist (2% each).

At present there are no specific interventions known to improve these pathogenic mechanisms; however, there is evidence to suggest that the organisation

of care could have an impact on the onset of foot disease. These patients therefore should have regular foot surveillance, education, and support from appropriate specialists to manage their foot care. All professionals involved in the care of patients with diabetes and renal disease should be aware of the LDK378 mw extent to which the patient’s feet are at risk. Copyright © 2012 John Wiley & Sons. “
“There have been few studies investigating the use of GLP-1 agonists in patients with insulin-treated type 2 diabetes and none looking at the costing. We compared the efficacy and relative cost of adding exenatide treatment to patients with type 2 diabetes receiving either oral hypoglycaemic agents (OHAs) or insulin. Patients were recruited from West Suffolk Hospital Diabetes Centre. Data were acquired retrospectively from 207 patients completing six months of treatment. Of 207 patients, 188 demonstrated good clinical progress with a mean HbA1c reduction of 1.6% (p<0.0001) and weight loss of 6.9kg (p<0.0001). Nineteen patients discontinued exenatide as HbA1c reduction did not achieve the NICE target (0.4%; p=0.29), but they did achieve significant weight loss (5.6kg; p<0.0001). The 188 patients continuing

on exenatide were sub-divided XL765 chemical structure into insulin-treated (n=88) or tablet-treated (n=100). At six months, tablet-treated patients achieved an HbA1c reduction of 1.6% (p<0.0001) and weight loss of 6.5kg (p<0.0001). Insulin-treated patients achieved similar results: HbA1c reduction 1.6% (p<0.0001), weight loss 7.3kg (p<0.0001). After six months of exenatide treatment, the mean reduction in daily insulin dose was 48 units/person in the insulin-treated group. In the tablet-treated group, the cost of diabetic medication (per person/month) after six months was £54.90 above baseline, whereas in the insulin-treated group this was only £36.20 above baseline, because the reduction Unoprostone in insulin dose offset the cost of exenatide. It was concluded that exenatide is clinically

effective in both insulin-treated and tablet-treated type 2 diabetes, but is more cost effective in those originally treated with insulin. Copyright © 2011 John Wiley & Sons. “
“The aims of this study were to determine whether the introduction of a diabetes management e-module can increase junior doctors’ confidence in managing inpatients with diabetes and contribute to improvements in patient care. A diabetes e-module was introduced at Barnet and Chase Farm Hospitals NHS Trust in October 2010. Junior doctors completed it and undertook an online exam at the end. Junior doctors were surveyed once, six to eight months after completing the e-module, and retrospectively ranked their confidence and knowledge levels in managing inpatients with diabetes before and after completing the e-module.

The practitioner’s recommendation was highly important Alvelestat order for 63% of travelers. Overall, fewer travelers in the At+Pro group (17%—14/84) compared to the Dxy group (34%—24/70) reported side effects. The majority of travelers in the Mfl group (55%—17/31) reported side effects. The most frequent side effects were gastrointestinal irritation, which occurred with all three antimalarials

and neuropsychiatric events relating to changes in mood and behavior—nearly all of which occurred in the Mfl group. Photosensitivity was also reported in 13% of the Dxy group. The primary aim of this study was to assess traveler’s perceptions of, and self-reported adherence to, antimalarial medication in those traveling to crPF zones from the UK. It is recognized that self-reported

adherence level are often higher than adherence measured by objective methods, as has been clearly demonstrated in a study examining mefloquine chemoprophylaxis.13 Although the data concerning the mefloquine group has been included, the failure to achieve sufficient recruitment into this arm is perhaps a reflection of the lack of popularity of this agent in the UK and that only shorter term travelers were included. A clear finding was that travelers prescribed At+Pro were more adherent to their medication than those prescribed Dxy. The situation with Mfl was less clear, partly due to the lack of statistical power, and partly due to the difference in results for median percentage and categorical measures of adherence. The latter was probably NVP-AUY922 mouse due to the once-weekly regime for Mfl which would mean that misreporting of tablet numbers would have a proportionately greater impact on results. The results suggest that overall adherence to Mfl was either similar to or better than Dxy and similar to At+Pro for categorical adherence, but further research would be needed to establish this. Self-reported adherence was high both pre- and during the period of travel and the main period for reported non-adherence appears to be in the post-travel period. Absolute compliance with

no missed doses was reported by 70%, compared to a recent study,14 where 89% of participants claimed complete adherence. The study Morin Hydrate did not compare adherence to any other antimalarial and was assessed by telephone interview. That self-reported adherence might be higher than actual adherence, has been shown in other studies.15 An investigation into prophylaxis to Mfl13 has also indicated that much of the poor adherence can be attributed to the post-travel period. The results from the present study showed that travelers on At+Pro were more than twice as likely to report taking all or at least 80% of their post-travel medication than travelers on Dxy. The most likely reason for this is the shorter post-travel dosing period for At+Pro, 1 week compared with four weeks for Dxy.

Two millilitre of venous blood was collected from each subject, using disposable syringes, and promptly transferred to a lidded glass vial. Before clotting could occur, the reagent ethylene

diamine tetra acetic acid, which binds to lead in blood and facilitates its separation at the next stage, was added in equal volume to the blood and the mixture was shaken for 2 min10. To prevent sample contamination with exogenous lead, all laboratory glassware was cleansed with detergent and double-distilled water; they were then immersed in a 2-m HNO3 overnight and washed several times with double-distilled water before a final rinse with deionized Selleck PD-332991 water1. Each tooth was cleaned and soaked in a 3% solution of hydrogen peroxide to remove organic material, after which it was washed several times with double-distilled water and deionized water, air dried and weighed. The tooth was then dissolved in 3 mL of 70% HNO3 and 1 mL of 70% perchloric acid (HClO4) in a 50-mL beaker. The mixture was heated slowly until a clear,

colourless solution was obtained, which was then evaporated until dry. The selleck products digest was then rinsed with distilled water, filtered if cloudy, made up to 10 mL and shaken1. The lead concentration in the final digested solution was determined by using Flame Atomic Absorption Spectrophotometer (AAS) with electrothermal atomization (Varian Inc., Palo Alto, CA, USA). The specifications of the instrument were: lamp current 9.0 mA, wavelength 217.0 nm, band pass 0.5–1.0 nm, ash temperature 800°C and atomization 2300°C without temperature

control1. The blood sample was mixed thoroughly by inverting the sample container 15 times. A 3-mL aliquot of the blood sample was immediately dispensed into a centrifuge tube. Ammonium Pyrrolidine Dithio Carbamate solution (0.5 mL) was P-type ATPase added to the tube, and the tube was capped and inverted 15 times. The tube was then allowed to stand for 5 min, after which 3 mL of n-butyl acetate was added to the tube. The tube was capped again and shaken for a minimum 3 min at a rate sufficient to ensure mixing of the organic layer and blood. The tube was then centrifuged at 3000 revolutions/min for 2 min. The organic layer was aspirated into the flame of the AAS and absorbance was recorded10,11. The values obtained were subjected to statistical analysis using the Statistical Package for Social Sciences (SPSS-15) software for windows. Group comparison between males and females was carried out by using the Student’s t-test. Analysis of variance was used to assess group comparison for tooth type, age, and village. A critical value of P < 0.05 was considered statistically significant. The present study was carried out to determine and correlate the lead levels in blood and teeth of 100 children, all residents of villages located in the vicinity of a zinc–lead smelter.

Amyloid fibrils are rich in β-sheet and can be observed with thioflavin

Alisertib cell line T (ThT) assay or by staining with Congo red, indicating that they contain a hydrophobic region. Although these fibrillar amyloids were previously considered to be the primary factor in the induction of pathology in these protein conformational diseases, recent studies indicate that small oligomers or protofibrils, rather than amyloid fibrils, may play an important role in cytotoxicity (Lesnéet al., 2006; Haataja et al., 2008). In this study, we compared TDH and TRH to investigate whether membrane toxicity by the toxins is induced by amyloidogenicity upon heating or small oligomerized tetrameric structures. TRH showed less amyloidogenicity compared with that of TDH. However, the hemolytic activity of TRH was similar to that of TDH. These data indicate that membrane disruption by the TDH family is mediated by tetrameric structures and not by the amyloidogenicity. We also compared

the circular dichroism (CD) spectra of TDH and TRH in the heat-denatured state and found that an incorrect CHIR-99021 order refolding process resulted in loss of the Arrhenius effect of TRH. Purification of recombinant TDH was performed as described previously (Naim et al., 2001). N-terminal signal peptide-deleted (1–24 amino acids) trh1 (GenBank accession no. AB112353) was inserted into the expression vector pET-28a (Novagen). For the expression of recombinant TRH, we transformed a plasmid vector pET28-a harboring trh1 gene into Escherichia coli JM109 (DE3) cells (Promega). The transformant was cultured in Luria–Bertani broth (1% Bacto tryptone, 0.5% yeast extract, and 1% NaCl) containing 100 μg mL−1 of kanamycin at 30 °C for 30 h with rotary shaking, and then centrifuged at 6000 g for 30 min. We added ammonium sulfate (55% saturation) to the supernatant and allowed it to stir overnight, followed by centrifugation at 10 000 g for 1 h. The pellet was Vorinostat cell line dissolved in 10 mM phosphate buffer

(pH 7.4) and dialyzed against the same buffer. We applied this solution to a series of columns: Cellulofine Hap (hydroxyapatite) (Seikagaku-Kogyo, Tokyo, Japan), Toyopearl DEAE-650M (Tosoh, Tokyo, Japan), Resource-Phe (Amersham Pharmacia Biotech AB, Uppsala, Sweden), and Superose 6 (GE Healthcare, Uppsala, Sweden). Hemolytic activities were measured as described previously (Fukui et al., 2005). Far-UV CD spectra were recorded with a J-720W spectropolarimeter (Jasco, Tokyo, Japan) equipped with a thermoelectric temperature controller. Data were analyzed with the software provided by Jasco. Measurements were taken in a quartz cuvette with a path length of 2 mm, scanned in steps of 0.2 nm at a rate of 50 nm min−1. Samples of 0.2 mg mL−1 TRH in 10 mM phosphate buffer (pH 7.4) were heated up from 37 to 90 °C at a heating rate of 0.1 °C min−1. After heat treatment at 90 °C, the temperature was decreased rapidly by 30 °C min−1 or slowly by 1 °C min−1 decrements to 37 °C.