间接测热法和反向Fick氏法测定心脏手术病人术后氧耗量的比较

目的 采用反向Fick氏法和间接测热法测定心脏手术后病人的全身氧耗,并比较两种测量方法的相关性和准确性。方法8例心脏手术术后病人,分别在入ICU后2h和6h时同时采用反向Fick氏和间接测热法测定病人的全身氧耗量。结果 间接测热法和反向Fick法测得氧耗分别是(16±30)ml·min-1·m-2和(127±23)ml·min-1·m-2,前者的测定结果显著高于后者(P<0.01)。相关分析显示两者有较好的相关性(r=0.92,P<0.01)。采用Bland和Altman统计分析提示两种测定结果的平均偏离值为(35.5±13.4)ml·min-1·m-2,其95％的分布范围是(9～62)ml·min-1·m-2,提示两种测量结果间的一致性较差。结论 两种方法测定心脏病人术后全身氧耗的结果有明显差异,其中反向Fick氏法的测定结果误差大、准确性差,而间接测热法是较好的临床选择。     

Indomethacin-Dipalmitoylphosphatidylcholine Interaction. A Calorimetric Study of Drug Release from Poly(Lactide-co-glycolide) Microspheres into Multilamellar Vesicles

A comparative study of indomethacin controlled release from poly(lactide-co-glycolide) (50:50, molecular weight 3000) (PLGA) microspheres loaded with two different amounts of drug (10.9 ± 1%, and 34.1 ± 1% w/w) and pure free indomethacin, considering the effects exerted by the drug on the thermotropic behavior of dipalmitoylphosphatidylcholine multilamellar vesicles, was carried out by differential scanning calorimetry (DSC). The release was monitored by comparing the effect exerted by the free indomethacin on lipid thermotropic behavior with that of the drug released by the microspheres and relating these effects to a lipid aqueous dispersion containing the molar ratio of drug able to cause it. By DSC measurements, the pure free indomethacin was found to be able to have a fluidifying effect on the model membrane, causing a shift toward lower values of the transitional temperature (T_m), characteristic of phospholipid liposomes, without variations in the enthalpic changes (ΔH). This shift was found to be modulated by the drug molar fraction with respect to the lipid concentration in the aqueous dispersion. Successively, calorimetric measurements were performed on suspensions of blank liposomes added to weighed amounts of unloaded and indometha-cin-loaded microspheres as well as free powdered indomethacin, and the T_m shifts of the lipid bilayer caused by the drug released from the polymeric system, as well as by the free drug, were compared with that caused by free drug increasing molar fractions dispersed directly on the membrane, employed as a calibration curve to obtain the fraction of drug released. This drug release model could be employed to determine the different kinetics involved in the drug transfer from the microspheres to a membrane. This in vitro study suggests that the kinetic process involved in drug release is influenced by the amount of drug loaded in the microspheres. This calorimetric study shows that the PLGA microspheres are a good delivery system able to sustain drug release. Moreover, the DSC technique applied to the drug interaction with biomembranes constitutes a good tool for determining the drug release representing an innovative alternative in vitro model.     

Behavioral and physiologic responses to caloric restriction in mice

The purpose of the review is to highlight the influences of ambient temperature (T(a)) and caloric restriction (CR) on metabolism, cardiovascular function and behavior in mice. Standard vivarium ambient temperatures (T(a)?23 degrees C) are a mild cold stress for mice requiring elevated metabolic rate and food intake. Increasing T(a) into the zone of thermoneutrality (TMN?29-33 degrees C) markedly reduces food intake, metabolic rate, heart rate (HR) and blood pressure in mice. Mice are members of a diverse, yet unique group of homeothermic animals that respond to thermal and energetic challenges by allowing body temperature (T(b)) to fall to less than 31 degrees C, a condition known as torpor. In mice housed at standard T(a), torpor is induced by a single night of fasting or a few days of CR. The mechanisms responsible for initiating torpor are related to reduced caloric availability, but do not require leptin. Mice housed at TMN and subjected to CR exhibit physiologic reductions in metabolic rate and HR, but do not appear to enter torpor. Finally, mice exhibit differential locomotor activity responses during CR that depends on T(a). At standard T(a), mice display increased light-phase home-cage activity with CR. This response is virtually eliminated when CR is performed at TMN. We suggest that researchers using mice to investigate energy homeostasis and cardiovascular physiology carefully consider the influence of T(a) on physiology and behavior.     

Real-time assessment of bacteriophage T3-derived antimicrobial activity against planktonic and biofilm-embedded Escherichia coli by isothermal microcalorimetry

Bacterial biofilms, highly resistant to the conventional antimicrobial therapy, remain an unresolved challenge pressing the medical community to investigate new and alternative strategies to fight chronic implant-associated infections. Recently, strictly lytic bacteriophages have been revalued as powerful agents to kill antibiotic-resistant bacteria even in biofilm. Here, the interaction of T3 bacteriophage and planktonic and biofilm Escherichia coli TG1, respectively, was evaluated using isothermal microcalorimetry. Microcalorimetry is a non-invasive and highly sensitive technique measuring growth-related heat production of microorganisms in real-time. Planktonic and biofilm E. coli TG1 were exposed to different titers of T3 bacteriophage, ranging from 10² to 10⁷ PFU/ml. The incubation of T3 with E. coli TG1 showed a strong inhibition of heat production both in planktonic and biofilm already at lower bacteriophage titers (10³ PFU/ml). This method could be used to screen and evaluate the antimicrobial potential of different bacteriophages, alone and in combination with antibiotics in order to improve the treatment success of biofilm-associated infections.     

Investigation of Solid Dosage Form Components Interaction Using Orthogonal Techniques- Discovery of New Forms of Sodium Stearyl Fumarate

Physical or chemical interactions between drug product (DP) components can occur during manufacturing and/or upon storage; and may alter DP shelf life and performance. In this work a new Powder X-ray Diffraction (PXRD) peak was observed in DP under accelerated storage conditions. Due to the complex drug product matrix (including API, polymer, fillers, super disintegrant and lubricant), it was challenging to pinpoint the component(s) responsible for the new peak. In addition to PXRD, other orthogonal techniques including Differential Scanning Calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor sorption (DVS), Solid State Nuclear Magnetic Resonance (SSNMR) and Infrared (IR) spectroscopy were employed in this investigation to understand the root cause mechanistically. Specifically, multi nuclei SSNMR (¹H, ²³Na, ¹³C) was instrumental in delineating the components of the matrix. We identified the root cause to be an acid base reaction occurring in the DP, whereby sodium ion in sodium stearyl fumarate (SSF) is replaced by proton leading to SSF form conversion. We also identified commercially available SSF to be a hydrate that can dehydrate to an anhydrous form upon heating. In general, the same techniques can be used to investigate interactions of any multi component solid dosage forms.     

New Insight Into Thermodynamical Stability of Carbamazepine

Carbamazepine (CBZ)—an antiepileptic drug—belongs to Biopharmaceutics Classification System II Class. It has low solubility and consequently limited bioavailability. One of the ways to improve drugs solubility is amorphization of their structure. Herein, cooling CBZ—at different cooling rates—was investigated as a way to obtain glassy, better soluble form. During preliminary differential scanning calorimetry experiments, some peculiar behavior of the examined material, different from those stated in the literature, was observed. Further investigations using differential scanning calorimetry, thermogravimetric analysis, and polarizing optical microscope revealed that decomposition temperature of CBZ is about 30°C lower than previously assumed. Moreover, high-resolution thermogravimetric measurements indicate that some decomposition processes could start even below the temperature reported as the melting point of the form I of CBZ.     

Comparative assessment of in vitro release kinetics of calcitonin polypeptide from biodegradable microspheres

The objective of our study was to compare the in vitro release kinetics of a sustained-release injectable microsphere formulation of the polypeptide drug, calcitonin (CT), to optimize the characteristics of drug release from poly-(lactide-co-glycolide) (PLGA) copolymer biodegradable microspheres. A modified solvent evaporation and double emulsion technique was used to prepare the microspheres. Release kinetic studies were carried out in silanized tubes and dialysis bags, whereby microspheres were suspended and incubated in phosphate buffered saline, sampled at fixed intervals, and analyzed for drug content using a modified Lowry protein assay procedure. An initial burst was observed whereby about 50% of the total dose of the drug was released from the microspheres within 24 hr and 75% within 3 days. This was followed by a period of slow release over a period of 3 weeks in which another 10-15% of drug was released. Drug release from the dialysis bags was more gradual, and 50% CT was released only after 4 days and 75% after 12 days of release. Scanning electron micrographs revealed spherical particles with channel-like structures and a porous surface after being suspended in an aqueous solution for 5 days. Differential scanning calorimetric studies revealed that CT was present as a mix of amorphous and crystalline forms within the microspheres. Overall, these studies demonstrated that sustained release of CT from PLGA microspheres over a 3-week period is feasible and that release of drug from dialysis bags was more predictable than from tubes.     

Solid-state transformation of a leukotriene antagonist

SK&F 104353, a potent leukotriene antagonist, exists in two physically distinct species as a disodium salt. Type I melts with decomposition, showing a single endotherm in the region of 250–270°C in differential scanning calorimetry (DSC) thermograms, as well as exhibiting a weak powder X-ray diffraction (XRD) pattern. Type II contains a single DSC melting endotherm in the same region, but in contrast has additional, smaller endotherms in the region of 70–85°C; the new transitions remain upon cooling and reheating. The powder XRD pattern of II is very similar to that of I with respect to peak positions, but II's bands are sharper, suggestive of increased crystallinity. Types I and II can also be distinguished using solution calorimetry, manifesting different heats of solution in an ethanol/water mixture. Apparent irreversible conversion of I to II can readily occur, and has a significant impact with respect to physical stability of potential suspension aerosol formulations. The results are explained in terms of a solid-state transformation, i.e., from a metastable, partially amorphous form of low crystallinity (I) to a similar but more physically stable form of higher crystallinity (II); the conversion is facilitated by the presence of water, although true ‘hydration’ is not involved. Temperature-dependent Raman spectroscopy has been applied to characterize the transformation in more detail. The unusual solid-state behavior of this compound may reflect more general structural characteristics of leukotriene antagonists.     

The role of extracellular sodium on heart muscle energetics

A study has been made of changing external sodium concentration [Na]_e, over the range 75 to 200 mmol · l^–1, on contractile parameters and heat production in isolated, arterially perfused, interventricular rabbit septa.-The observed changes in maximum rate of contraction with [Na]_e, either in the presence of a constant external Ca concentration [Ca]_e or in the presence of a constant [Na] _e ² /[Ca]_e ratio, paralleled those observed for tension development (T). On the other hand the maximal rate of relaxation and the ratio increased. While the ratio between active heat production and developed tension remained unaltered (0.111±0.003 mJ · mN^–1 · g^–1 dry weight), resting heat production increased with [Na] _e ² with a slope of 95±18 mW · g^–1 · mol^–2 · l². Under resting conditions, a decrease in [Na]_e of 50 mmol · l^–1 induced a fall in⁴²K uptake of about 16 nmol · s^–1 · g^–1 without changes in⁴²K efflux, suggesting that such an intervention depresses K influx. If the depressed K influx, induced by a decrease in [Na]_e of 50 mmol · l^–1 is associated with a decrease in Na–K pump activity, a fall in resting heat production of about 0.64 mW · g^–1 would be expected. This represent 56% of the calculated change in the resting heat production, 1.14±0.22 mW · g^–1 (mean ± one confidence interval), suggesting that some process in addition to a depressed Na-K pump activity may be altered by changes in [Na]_e.Supported by the Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET), Argentina     

A calorimetric study on diflunisal release from poly(lactide-co-glycolide) microspheres by monitoring the drug effect on dipalmitoylphosphatidylcholine liposomes: temperature and drug loading influence

Diflunisal release from poly-Lactide-co-Glycolide (50:50, 34,000 MW) microspheres loaded with two different amounts of drug (2.5 +/- 0.5% and 10 +/- 0.5% w/w) was monitored by following the effects exerted by the drug on the thermotropic behavior of dipalmitoylphosphatidylcholine unilamellar vesicles at different temperatures. The effects of the drug released from the microspheres on the thermotropic behavior of lipid aqueous dispersion containing different molar ratios of drug was detected by differential scanning calorimetry and was compared with the effects exerted by the free Diflunisal. Diflunisal affects mainly the temperature (T_m) of the transition characteristic of phospholipid vesicles as model biomembrane, causing a shift toward lower values. This shift was modulated by the drug molar fraction with respect to the lipid concentration in the aqueous dispersion. Afterward, calorimetric measurements were performed on suspensions of blank liposomes added to weighed amounts of unloaded and differently Diflunisal-loaded microspheres as well as free powdered Diflunisal after incubation for increasing times at three different temperatures (25, 37, and 50°C). The T_m shifts of the lipid bilayer, caused by the drug released from polymeric system as well as by the free drug during incubation periods, were compared with that caused by free drug increasing molar fractions dispersed directly on the membrane, employed as a calibration curve to obtain the fraction of drug released. This in vitro study suggests that the kinetic process involved in drug release is influenced by the amount of drug loaded in the microspheres as well as by the temperature acting on drug solubility and membrane disorder. This drug release model, monitored by the calorimetric technique shows that a) the poly-Lactide-co-Glycolide microspheres are a good delivery system able to sustain the drug release; b) the differential scanning calorimetry technique applied on the drug interaction with biomembranes constitutes a good tool to follow the drug release; 3) this model, representing an innovative alternative in vitro model, should be used to determine the different kinetics involved in the drug transfer from a drug delivery system to a membrane as uptake site.