Trapping by amylose of the aliphatic chain grafted onto chlorogenic acid: importance of the graft position.

5-Caffeoylquinic acid (chlorogenic acid), is classified in acid-phenols family and as polyphenolic compounds it possesses antioxidant activity. The oxydative modification of chlorogenic acid in foods may lead to alteration of their qualities; to counteract these degradation effects, molecular encapsulation was used to protect chlorogenic acid. Amylose can interact strongly with a number of small molecules, including lipids. In order to enable chlorogenic acid complexation by amylose, a C16 aliphatic chain was previously grafted onto the cycle of quinic acid. This work showed that for the two lipophilic derivatives of chlorogenic acid: hexadecyl chlorogenate obtained by alkylation and 3-O-palmitoyl chlorogenic acid obtained by acylation; only the 3-O-palmitoyl chlorogenic acid complexed amylose. The chlorogenic acid derivatives were studied by X-ray diffraction, differential scanning calorimetry and NMR to elucidate the interaction. By comparing the results with previous work on the complexation of amylose by 4-O-palmitoyl chlorogenic acid, the importance of the aliphatic chain position on the cycle of the quinic acid is clearly highlighted. A study in molecular modeling helped to understand the difference in behavior relative to amylose of these three derivatives of chlorogenic acid.

Cerebral haemodynamic physiology during steep Trendelenburg position and CO(2) pneumoperitoneum.

BACKGROUND: The steep (40°) Trendelenburg position optimizes surgical exposure during robotic prostatectomy. The goal of the current study was to elucidate the influence of this patient positioning on cerebral blood flow and zero flow pressure (ZFP), and to assess the validity of different methods of evaluating ZFP. METHODS: In 21 consecutive patients who underwent robotic endoscopic radical prostatectomy under general anaesthesia, transcranial Doppler flow velocity waveforms and invasive arterial and central venous pressure (CVP) waveforms suitable for analysis were recorded throughout the whole operative procedure in 14. The ZFP was determined by regression analysis of the pressure-flow plot and by different simplified formulas. The effective cerebral perfusion pressure (eCPP), pulsatility index (PI), and resistance index (RI) were determined. RESULTS: While patients were in the Trendelenburg position, the ZFP increased in parallel with the CVP. The PI, RI, gradient between the ZFP and CVP, and the gradient between the CPP and the eCPP did not increase significantly (P<0.05) after 3 h of the steep Trendelenburg position. Using the formula described by Czosnyka and colleagues, the ZFP correlated closely with that calculated by linear regression throughout the course of the operation. CONCLUSIONS: Prolonged steep Trendelenburg positioning and CO(2) pneumoperitoneum does not compromise cerebral perfusion. ZFP and eCPP are reliable variables for assessing brain perfusion during prolonged steep Trendelenburg positioning.

Coasting: worth the effort?

A new anesthesia machine incorporates a "coasting mode", but the extent to which a coasting technique can maintain anesthesia at the end of a procedure under optimal conditions (closed circuit anesthesia) remains unknown. Sixty-nine patients undergoing peripheral or abdominal surgery were assigned to 1 of 9 groups, depending on when desflurane coasting (in O2/air) was started (after 4, 9, 16, 25, 36, 49, 64, 81, or 100 min). The end-expired desflurane concentration was maintained at 4.5% in O2/air prior to coasting with a conventional anesthesia machine. After initiating coasting (using a closed-circuit technique), we examined when the end-expired desflurane concentration reached 70, 60, 50, and 40% of its value during maintenance (= 30, 40, 50 and 60% decrement times, respectively). Decrement times increased with increasing duration of anesthesia, and varied widely. After 64 min of maintenance anesthesia, the end-expired desflurane concentration remained at or above 70, 60, 50, and 40% of its maintenance value during 10.3 +/- 2.3, 16.0 +/- 3.5, 25.0 +/- 5.9, and 45.4 +/- 19.3 min, respectively (average +/- standard deviation). Coasting can briefly maintain anesthesia towards the end of a procedure. While savings with an automated coasting mode are likely to be modest per patient, they may become substantial when multiplied by the number of procedures per day per operating room with no increase in the clinical workload of the anesthesia provider.

Effect of the mode of administration of inhaled anaesthetics on the interpretation of the F(A)/F(I) curve--a GasMan simulation.

The effects of blood solubility, cardiac output and ventilation on the rise of the alveolar towards the inspired concentration, the F(A)/F(I) curve, of an inhaled anaesthetic are often thought to reflect how these factors affect wash-in of the central nervous system compartment and, therefore, speed of induction because F(A) is the partial pressure ultimately attained in the central nervous system (F(VRG)). These classical F(A)/F(I) curves assumed a constant F(I). We used GasMan to examine whether changes in solubility, cardiac output and ventilation affect the relationship between the F(A)/F(I) curve and F(VRG) differently while either F(I) or F(A) are kept constant. Using GasMan, we studied the effects of solubility (desflurane vs isoflurane), cardiac output (5 vs. 10 l x min(-1)) and minute ventilation (4 vs. 8 l x min(-1)) on F(A), F(I), F(A)/F(I) and F(VRG) with either F(I) kept constant or F(A) kept constant (at 1 minimum alveolar concentration). High fresh gas flows were used to avoid rebreathing, so that the delivered concentration matched F(I). Despite similar effects on the F(A)/F(I) curve, the effects on F(VRG) differed. With constant F(I), lower solubility or higher ventilation results in a higher F(VRG) and a higher cardiac output results in a lower F(VRG). With constant F(A), solubility has only a minimal effect on F(VRG); an increase in cardiac output hastens the rise of F(VRG) to the same plateau value; and a change in ventilation has minimal effect on F(VRG). Despite similar effects on the F(A)/F(I) curve, the effects of solubility, cardiac output and ventilation on the F(VRG) are different when either F(I) or F(A) are kept constant. With the F(I) kept constant, induction of anaesthesia is slower with a higher cardiac output, but with F(A) kept constant, induction of anaesthesia is faster with a higher cardiac output. The introduction of an end-expired closed-loop feedback administration of inhaled anaesthetics makes this distinction clinically relevant.

Involvement of T-cell receptor-beta alterations in the development of otosclerosis linked to OTSC2.

Otosclerosis is a common form of hearing loss, characterized by disordered bone remodeling in the otic capsule. Within the otosclerotic foci, several immunocompetent cells and immune-modulating factors can be found. Different etiological theories involving the immune system have been suggested. However, a genetic component is clearly present. In large otosclerosis families, seven autosomal-dominant loci have been found, but none of the disease-causing genes has been identified. This study focused on the exploration of the second otosclerosis locus on chromosome 7q34-36 (OTSC2), holding the T-cell receptor beta locus (TRB locus). A significantly lower T-cell receptor-beta (TCR-beta) mRNA expression and percentage of blood circulating TCR-alphabeta(+) T cells was detected in OTSC2 patients compared with controls and patients with the complex form of the disease. Further analysis illustrated more significant disturbances in specific T-cell subsets, including an increased CD28(null) cell population, suggesting a disturbed T-cell development and ageing in OTSC2 patients. These disturbances could be associated with otosclerotic bone remodeling, given the known effects of immunocompetent cells on bone physiology. These data implicate the TRB locus as the causative gene in the OTSC2 region and represent an important finding in the elucidation of the disease pathology.

Influence of steep Trendelenburg position and CO(2) pneumoperitoneum on cardiovascular, cerebrovascular, and respiratory homeostasis during robotic prostatectomy.

BACKGROUND: The steep (40 degrees ) Trendelenburg position optimizes surgical exposure during robotic prostatectomy. The goal of the current study was to investigate the combined effect of this position and CO(2) pneumoperitoneum on cardiovascular, cerebrovascular, and respiratory homeostasis during these procedures. METHODS: Physiological data were recorded during the whole surgical procedure in 31 consecutive patients who underwent robotic endoscopic radical prostatectomy under general anaesthesia. Heart rate, mean arterial pressure, central venous pressure, Sp(o(2)), Pe'(co(2)), P(Plat), tidal volume, compliance, and minute ventilation were monitored and recorded. Arterial samples were obtained to determine the arterial-to-end-tidal CO(2) tension gradient. Continuous regional cerebral tissue oxygen saturation (Sct(o(2))) was determined by near-infrared spectroscopy. RESULTS: Although patients were in the Trendelenburg position, all variables investigated remained within a clinically acceptable range. Cerebral perfusion pressure (CPP) decreased from 77 mm Hg at baseline to 71 mm Hg (P=0.07), and Sct(o(2)) increased from 70% to 73% (P<0.001). Pe'(co(2)) increased from 4.12 to 4.79 kPa (P<0.001) and the arterial-to-Pe'(co(2)) tension difference increased from 1.06 kPa in the normal position to a maximum of 1.41 kPa (P<0.001) after 2 h in the Trendelenburg position. CONCLUSIONS: The combination of the prolonged steep Trendelenburg position and CO(2) pneumoperitoneum was well tolerated. Haemodynamic and pulmonary variables remained within safe limits. Regional cerebral oxygenation was well preserved and CPP remained within the limits between which cerebral blood flow is usually considered to be maintained by cerebral autoregulation.

Derivation and prospective testing of a two-step sevoflurane-O2-N2O low fresh gas flow sequence.

Simple vaporiser setting (F(D)) and fresh gas flow (FGF) sequences make the practice of low-flow anaesthesia not only possible but also easy to achieve. We sought to derive a sevoflurane F(D) sequence that maintains the end-expired sevoflurane concentration (F(A)sevo) at 1.3% using the fewest possible number of F(D) adjustments with a previously described O2-N2O FGF sequence that allows early FGF reduction to 0.7 l min(-1). In 18 ASA physical status I to IH patients, F(D) was determined to maintain F(A)sevo at 1.3% with 2 l min(-1) O2 and 4 l min(-1) N2O FGF for three minutes, and with 0.3 and 0.4 l min(-1) thereafter. Using the same FGF sequence, the F(D) schedule that approached the 1.3% F(A)sevo pattern with the fewest possible adjustments was prospectively tested in another 18 patients. The following F(D) sequence approximated the F(D) course well: 2% from zero to three minutes, 2.6% from three to 15 minutes and 2.0% after 15 minutes. When prospectively tested, median (25th; 75th percentile) performance error was 0.8 (-2.9; 5.9)%, absolute performance error 6.7 (3.3; 10.6)%, divergence 18.2 (-5.6; 27.4)%.h(-1) and wobble 4.4 (1.7; 8.1) %. In one patient, FGF had to be temporarily increased for four minutes. One O2/N2O rotameter FGF setting change from 6 to 0.7 l min(-1) at three minutes and two sevoflurane F(D) changes at three and 15 minutes maintained predictable anaesthetic gas concentrations during the first 45 minutes in all but one patient in our study.

Desflurane consumption with the Zeus during automated closed circuit versus low flow anesthesia.

INTRODUCTION: During automated closed-circuit anesthesia (CCA), the Zeus (Dräger, Lübeck, Germany) uses a high initial fresh gas flow (FGF) to rapidly attain the desired agent and carrier gas concentrations, resulting in a desflurane consumption well above patient uptake. Because both FGF and carrier gas composition can affect consumption, we determined the Zeus' agent consumption with automated CCA and with automated low flow anesthesia (LFA) (= maintenance FGF of 0.7 L min(-1)) with 3 different carrier gases. METHODS: After IRB approval, 65 ASA PS I or II patients undergoing general surgery received desflurane in either O2, O2/air, or O2/N2O, with the Zeus to maintain the end-expired concentration (FA) at 6, 6, and 4% and the F1O2 at 1.0, 0.6, and 0.4, respectively. In addition, patients were assigned to either automated CCA (O2 n = 11; O2/air n = 11; O2/N2O n = 11) or automated LFA (selected FGF 0.7 L min(-1)) (O2 n = 12; O2/air n = 11; O2/N2O n = 9). Demographics and desflurane consumption at 2, 4, 6, 8, 10, 20, 30, 40 and 50 min were compared. RESULTS: With the same carrier gas, desflurane consumption was lower with the CCA mode than with LFA mode after 4 min in the O2 groups, 6 min in the O2/air groups, and 30 min in the O2/N2O groups. Within each mode, desflurane consumption in the O2 and O2/air groups was identical at all times. Despite the use of a lower FA in the N2O groups, initial desflurane consumption was higher than in the O2 and O2/air groups, but it was lower later (> or = 15 min) only with LFA. DISCUSSION: After 50 min, desflurane consumption with automated CCA is lower than with automated LFA. However, initial agent consumption is complex, and N2O in particular may increase initial desflurane consumption (though ultimately resulting in lower desflurane usage because of its MAC sparing effect) because initial FGF is increased to rapidly reach the target concentrations. Differences in desflurane consumption only become apparent after FGF has stabilized to the target FGF.

Pulmonary gas exchange is well preserved during robot assisted surgery in steep Trendelenburg position.

INTRODUCTION: During robot assisted hysterectomies and prostatectomies, surgical exposure demands the application of a CO2 pneumoperitoneum with a very steep Trendelenburg position (40 degrees). The extent to which oxygenation and ventilation might be compromised intra-operatively remains poorly documented. METHODS: Dead-space ventilation and venous admixture were determined in 18 patients undergoing robot assisted hysterectomy (n = 6) or prostatectomy (n = 12). Anesthesia was maintained with desflurane in O2 or O2/air, with the inspired O2 fraction left at the discretion of the attending anesthesiologist. Controlled mechanical ventilation was used, but 15 min after assuming the Trendelenburg position and up until resuming the supine position pressure controlled ventilation was used. Dead-space ventilation and venous admixture were determined using Bohr's formula and Nunn's iso-shunt diagram, respectively, at the following 7 stages of the procedure: 15 min after induction; 5 min after applying the CO2 pneumoperitoneum (intra-abdominal pressure 12 mm Hg) but while still supine; 5, 60, and 120 min after assuming the Trendelenburg positioning; and 5 and 15 min after reassuming the supine position. RESULTS: Venous admixture did not change. Dead-space ventilation increased after Trendelenburg positioning, and returned to baseline values after resuming the supine position. However, individual patterns varied widely. DISCUSSION: The lung has a remarkable yet incompletely understood capacity to withstand the effects of a CO2 pneumoperitoneum and steep Trendelenburg position during general anesthesia. While individual responses vary and should be monitored, effects on dead-space ventilation and venous admixture are small and should not be an obstacle to provide optimal surgical exposure during robot assisted prostatectomy or hysterectomy.

Special aspects of pharmacokinetics of inhalation anesthesia.

Recent interest in the use of low-flow or closed circuit anesthesia has rekindled interest in the pharmacokinetics of inhaled anesthetics. The kinetic properties of inhaled anesthetics are most often modeled by physiologic models because of the abundant information that is available on tissue solubilities and organ perfusion. These models are intuitively attractive because they can be easily understood in terms of the underlying anatomy and physiology. The use of classical compartment modeling, on the other hand, allows modeling of data that are routinely available to the anesthesiologist, and eliminates the need to account for every possible confounding factor at each step of the partial pressure cascade of potent inhaled agents. Concepts used to describe IV kinetics can readily be applied to inhaled agents (e.g., context-sensitive half-time and effect site concentrations). The interpretation of the F(A)/F(I) vs time curve is expanded by reintroducing the concept of the general anesthetic equation-the focus is shifted from "how F(A) approaches F(I)" to "what combination of delivered concentration and fresh gas flow (FGF) can be used to attain the desired F(A)." When the desired F(A) is maintained with a FGF that is lower than minute ventilation, rebreathing causes a discrepancy between the concentration delivered by the anesthesia machine (=selected by the anesthesiologist on the vaporizer, F(D)) and that inspired by the patient. This F(D)-F(I) discrepancy may be perceived as "lack of control" and has been the rationale to use a high FGF to ensure the delivered matched the inspired concentration. Also, with low FGF there is larger variability in F(D) because of interpatient variability in uptake. The F(D)-F(I) discrepancy increases with lower FGF because of more rebreathing, and as a consequence the uptake pattern seems to be more reflected in the F(D) required to keep F(A) constant. The clinical implication for the anesthesiologist is that with high FGF few F(D) adjustments have to be made, while with a low FGF F(D) has to be adjusted according to a pattern that follows the decreasing uptake pattern in the body. The ability to model and predict the uptake pattern of the individual patient and the resulting kinetics in a circle system could therefore help guide the anesthesiologist in the use of low-flow anesthesia with conventional anesthesia machines. Several authors have developed model-based low FGF administration schedules, but biologic variability limits the performance of any model, and therefore end-expired gas analysis is obligatory. Because some fine-tuning based on end-expired gas analysis will always be needed, some clinicians may not be inclined to use very low FGF in a busy operating room, considering the perceived increase in complexity. This practice may be facilitated by the development of anesthesia machines that use closed circuit anesthesia (CCA) with end-expired feedback control--they "black box" these issues (see Chapter 21). In this chapter, we first explore how and why the kinetic properties of intravenous and inhaled anesthetics have been modeled differently. Next, we will review the method most commonly used to describe the kinetics of inhaled agents, the F(A)/F(I) vs time curve that describes how the alveolar (F(A)) approaches the inspired (F(I)) fraction (in the gas phase, either "fraction," "concentration," or "partial pressure" can be used). Finally, we will reintroduce the concept of the general anesthetic equation to explain why the use of low-flow or closed circuit anesthesia has rekindled interest in the modeling of pharmacokinetics of inhaled anesthetics. Clinical applications of some of these models are reviewed. A basic understanding of the circle system is required, and will be provided in the introduction.

A new, easy, and rapid high-throughput detection method for the common GJB2 (CX26), 35delG mutation.

GJB2 (Gap Junction protein beta type 2; Connexin 26, CX26) is known for its contribution to nonsyndromic recessive deafness (NSRD). One particular mutation, 35delG, a deletion of one guanine from a stretch of six leading to a frame shift early in the gene, has a high prevalence in populations from European descent. 35delG testing therefore has become a standard test in genetic diagnostic laboratories. Most of the currently available methods for the detection of 35delG are relatively time consuming, and not suited for high-throughput diagnostic testing. Within this paper we present a real-time PCR genotyping assay based on melting curve analysis, requiring only a single preparation step before the actual analysis. The assay was optimized on a panel of 48 samples with known 35delG genotypes and subsequently tested using a large Belgian population (N = 460) with unknown 35delG status. For the latter set of samples, real-time PCR results were validated with SNAPShot, an assay used in our laboratory for diagnostic purposes. The real-time PCR genotyping method has proven to be highly reliable, rapid, cost-effective, and suitable for high-throughput screening. We believe that this genetic test for 35delG will find widespread applications in the DNA diagnostic field.

Contribution of the N-acetyltransferase 2 polymorphism NAT2*6A to age-related hearing impairment.

BACKGROUND: Age-related hearing impairment (ARHI) is the most common sensory impairment in older people, affecting 50% of those aged 80 years. The proportion of older people is increasing in the general population, and as a consequence, the number of people affected with ARHI is growing. ARHI is a complex disorder, with both environmental and genetic factors contributing to the disease. The first studies to elucidate these genetic factors were recently performed, resulting in the identification of the first two susceptibility genes for ARHI, NAT2 and KCNQ4. METHODS: In the present study, the association between ARHI and polymorphisms in genes that contribute to the defence against reactive oxygen species, including GSTT1, GSTM1 and NAT2, was tested. Samples originated from seven different countries and were combined into two test population samples, the general European population and the Finnish population. Two distinct phenotypes for ARHI were studied, Z(low) and Z(high), representing hearing in the low and high frequencies, respectively. Statistical analysis was performed for single polymorphisms (GSTM1, GSTT1, NAT2*5A, NAT2*6A, and NAT2*7A), haplotypes, and gene-environment and gene-gene interactions. RESULTS: We found an association between ARHI and GSTT1 and GSTM1 in the Finnish population sample, and with NAT2*6A in the general European population sample. The latter finding replicates previously published data. CONCLUSION: As replication is considered the ultimate proof of true associations in the study of complex disorders, this study provides further support for the involvement of NAT2*6A in ARHI.

Prevalence of tinnitus and audiometric shape.

OBJECTIVES: Studies of tinnitus are often conducted on patient populations presenting for treatment. It is, however, difficult to generalise prevalence numbers and aetiological results from these studies to a healthy, elderly population. The first aim of our study was to determine the prevalence of tinnitus in an otologically screened population between 55 and 65 years old. Secondly, both prevalence and the specific characteristics of tinnitus were compared in subjects with either a flat audiogram, a high-frequency gently sloping audiogram or a high-frequency steeply sloping audiogram. METHODS: 1147 subjects (549 males and 598 females) were recruited through population registers and underwent thorough clinical and audiological examinations. Subjects who reported tinnitus in the general questionnaire about medical history and environmental exposure were invited to complete an additional questionnaire on tinnitus history. RESULTS: The prevalence of tinnitus was 19.3% according to the general questionnaire on medical health and environmental exposure and 11.8% according to the additional detailed tinnitus-specific questionnaire. Furthermore, our results indicate that gender has a significant effect (tinnitus is more common in males than in females), as does audiometric configuration (tinnitus is more common in subjects with a high-frequency steeply sloping audiogram than in subjects with a flat audiogram). Both effects were significant in noise-/solvent-exposed subjects, as well as in non-exposed subjects. Finally, comparison of "tinnitus characteristics" in subjects categorised by audiogram configuration revealed significant differences in loudness, pitch, temporal variability and family history of tinnitus.

Familial aggregation of tinnitus: a European multicentre study.

INTRODUCTION AND AIM: Tinnitus is a common condition affecting approximately 20% of the older population. There is increasing evidence that changes in the central auditory system following cochlear malfunctioning are responsible for tinnitus. To date, few investigators have studied the influence of genetic factors on tinnitus. The present report investigates the presence of a familial effect in tinnitus subjects. METHODS: In a European multicentre study, 198 families were recruited in seven European countries. Each family had at least 3 siblings. Subjects were screened for causes of hearing loss other than presbyacusis by clinical examination and a questionnaire. The presence of tinnitus was evaluated with the question "Nowadays, do you ever get noises in your head or ear (tinnitus) which usually last longer than five minutes". Familial aggregation was tested using three methods: a mixed model approach, calculating familial correlations, and estimating the risk of a subject having tinnitus if the disorder is present in another family member. RESULTS: All methods demonstrated a significant familial effect for tinnitus. The effect persisted after correction for the effect of other risk factors such as hearing loss, gender and age. The size of the familial effect is smaller than that for age-related hearing impairment, with a familial correlation of 0.15. CONCLUSION: The presence of a familial effect for tinnitus opens the door to specific studies that can determine whether this effect is due to a shared familial environment or the involvement of genetic factors. Subsequent association studies may result in the identification of the factors responsible. In addition, more emphasis should be placed on the effect of role models in the treatment of tinnitus.

The absorption, metabolism, and excretion of the novel neuromodulator RWJ-333369 (1,2-ethanediol, [1-2-chlorophenyl]-, 2-carbamate, [S]-) in humans.

RWJ-333369 (1,2-ethanediol, [1-2-chlorophenyl]-, 2-carbamate, [S]-; CAS Registry Number 194085-75-1) is a novel neuromodulator in clinical development for the treatment of epilepsy. To study the disposition of RWJ-333369, eight healthy male subjects received a single oral dose of 500 mg of (14)C-RWJ-333369. Urine, feces, and plasma were collected for analysis for up to 1 week after dosing. Radioactivity was mainly excreted in urine (93.8 +/- 6.6%) and much less in feces (2.5 +/- 1.6%). RWJ-333369 was extensively metabolized in humans, since only low amounts of parent drug were excreted in urine (1.7% on average) and feces (trace amounts). The major biotransformation pathways were direct O-glucuronidation (44% of the dose), and hydrolysis of the carbamate ester followed by oxidation to 2-chloromandelic acid, which was subsequently metabolized in parallel to 2-chlorophenyl glycine and 2-chlorobenzoic acid (mean percentage of the dose for the three acids together was 36%). Other routes were chiral inversion followed by O-glucuronidation (11%), and aromatic hydroxylation in combination with sulfate conjugation (5%). In plasma, unchanged drug accounted for 76.5% of the total radioactivity, with the R-enantiomer and the O-glucuronide of the parent drug as the only measurable plasma metabolites. With the use of very sensitive liquid chromatography-tandem mass spectrometry techniques, only traces of aromatic (pre)mercapturic acid conjugates were detected in urine (each <0.3% of the dose), suggesting a low potential for reactive metabolite formation. In conclusion, the disposition of RWJ-333369 in humans is characterized by virtually complete absorption, extensive metabolism, and unchanged drug as the only significant circulating species.