Optimization of Salmonella enteritidis recombinant heat shock protein 60 production.

The aim of the study was to optimize conditions for producing Salmonella Enteritidis recombinant heat shock protein 60 (rHsp60). Seven Escherichia coli host strains (Rosetta, Turner, C41, C43, Origami, BL21pLys, Rosetta pLys) were transformed by a recombinant plasmid containing Hsp60 gene from Salmonella Enteritidis, and then cultured and induced by isopropyl-beta-D-thiogalactopyranoside (IPTG). The highest S. Enteritidis rHsp60 yield was obtained using E. coli strain C41. Induction of this strain using IPTG allowed the yield 400 microg of S. Enteritidis Hsp60 protein/2L of culture, but by autoinduction the yield exceeded 800 microg/2L.

Experimental Studies and Theoretical Predictions for the H + D2 rarr > HD + D Reaction.

The H + H(2) exchange reaction constitutes an excellent benchmark with which to test dynamical theories against experiments. The H + D(2) (vibrational quantum number v = 0, rotational quantum number j = 0) reaction has been studied in crossed molecular beams at a collision energy of 1.28 electron volts, with the use of the technique of Rydberg atom time-of-flight spectroscopy. The experimental resolution achieved permits the determination of fully rovibrational state-resolved differential cross sections. The high-resolution data allow a detailed assessment of the applicability and quality of quasi-classical trajectory (QCT) and quantum mechanical (QM) calculations. The experimental results are in excellent agreement with the QM results and in slightly worse agreement with the QCT results. This theoretical reproduction of the experimental data was achieved without explicit consideration of geometric phase effects.

The relationship between concentrations of vitamin D-binding protein (DBP) in serum and colostrum of mares and in serum of their foals in the neonatal period.

Vitamin D-binding protein (DBP) participates in the actin scavenger system, it is a carrier of vitamin D and its derivatives, it manifests the capacity to bind mainly monounsaturated and saturated fatty acids, it binds to the surface of several cells and enhances chemotactic activity of C5a of the complement. The present study was aimed at answering the question whether serum DBP level in mares is related to levels of this protein in colostrum and in serum of its progeny. For this purpose, sera from 77 mares, colostra from 72 mares and sera from 69 Thoroughbred foals were collected. Mother's age, number of deliveries experienced in the past, month of delivery, feeding of foals with colostra were recorded. Blood of the foals was sampled from the umbilical vein during delivery (0h) and 36-48 h after delivery from the external jugular vein, colostra of the mares were obtained after delivery and blood of the mares was sampled 36-48 h after delivery. Concentration of DBP was estimated by a self-designed ELISA. In the present study, DBP concentrations in newborn's serum were found independent of their concentrations in mother's serum, her age and number of parities experienced in the past. Colostrum DBP level was found to be lower than that in the mare's serum and was not correlated to the concentration of this protein in mare's serum. There was no effect of colostrum feeding on DBP level in the foal serum. These results indicate that serum DBP concentration in newborn foals depends on factors which act directly on the foal. Because of the lack of correlation between plasma and colostrum concentrations of DBP, it can be assumed that DBP is synthesised in the mammary gland and/or specific transport mechanisms exist in the mammary gland.

Relationship Between the Skin Surface Temperature Changes During Sprint Interval Testing Protocol and the Aerobic Capacity in Well-Trained Cyclists.

The study investigated whether changes in body surface temperature in a sprint interval testing protocol (SITP) correlated with aerobic capacity in cyclists. The study involved 21 well-trained cyclists. Maximal aerobic power and maximal oxygen uptake relative to lean body mass (LBM-P(max) and LBM-VO(2max), respectively) were determined by incremental exercise testing on a cycle ergometer. SITP was administered 48 hours later and involved four 30-s maximal sprints interspersed with 90-s active recovery. Body surface temperature was recorded at the temple and arm and the delta difference between baseline temperature and temperature measured immediately after the first sprint (DeltaTt(1) and DeltaTa(1), respectively) and 80 seconds after the fourth sprint (DeltaTt(4) and DeltaTa(4)), respectively) was calculated. Significant correlations were found between DeltaTt4 and LBM-Pmax and LBM-VO(2max) (r=0.63 and r=0.75, respectively) with no significant change in DeltaTa(1) or DeltaTa(4). Body surface temperature, measured at the temple region, can be used to indirectly assess aerobic capacity during maximal sprint exercise.

Time of VO(2)max plateau and post-exercise oxygen consumption during incremental exercise testing in young mountain bike and road cyclists.

The purpose of this study was to compare markers of glycolytic metabolism in response to the Wingate test and the incremental test in road and mountain bike cyclists, who not different performance level and aerobic capacity. All cyclists executed the Wingate test and incremental test on a cycle ergometer. Maximal power and average power were determined during the Wingate test. During the incremental test the load was increased by 50 W every 3 min, until volitional exhaustion and maximal aerobic power (APmax), maximal oxygen uptake (VO2max), and time of VO(2)max plateau (Tplateau) were determined. Post-exercise measures of oxygen uptake (VO(2)post), carbon dioxide excretion, (VCO(2)post), and the ratio between VCO(2)/VO(2) (RERpost) were collected for 3 min immediately after incremental test completion. Arterialized capillary blood was drawn to measure lactate (La-) and hydrogen (H+) ion concentrations in 3 min after each test. The data demonstrated significant differences between mountain bike and road cyclists for Tplateau, VO(2)post, VCO(2)post, La- which was higher-, and RERpost which was lower-, in mountain bike cyclists compare with road cyclists. No differences were observed between mountain bike and road cyclists for APmax, VO(2)max, H(+) and parameters measured in the Wingate test. Increased time of VO2max plateau concomitant to larger post-exercise La- and VO(2) values suggests greater anaerobic contribution during incremental testing efforts by mountain bike cyclists compared with road cyclists.

Comparison of Mechanical Circulatory Support by the Use of Pulsatile Left Ventricular Assist Devices Polvad MEV and Continuous Flow Heart Ware and Heart Mate II in a Single-Center Experience.

BACKGROUND: Mechanical circulatory support is increasingly used in patients with heart failure as a bridge to transplant or recovery. Results of use the Polish POLVAD MEV pulsatile circulatory support system and its comparison with novel devices never was done. We compared the course of patients with left ventricular circulatory support (left ventricular assist device [LVAD]) supported by POLVAD MEV or continuous flow devices Heart Mate II (HM II) and Heart Ware (HW) in single-center cohort. METHODS: We retrospectively reviewed 44 patients who underwent Polvad Mev (group P; n = 24 [21M/3F]) or HW or HM II (group C; n = 20 [20M/0F]) implantation between April 2007 and February 2014. Patients were in INTERMACS 1 (6 in group P and 1 in group C) or 2. Preimplant demographics, and perioperative and postoperative clinical outcomes were reviewed between groups. We analyzed baseline signs of heart failure, comorbidities, complications, and the 30- and 90-day results. RESULTS: Among the groups, age, gender, weight, and cause of heart failure were comparable. Patients in group C suffered more frequently from hypercholesterolemia preoperatively. Patients in group P had more pulmonary complications (7 vs 0) after LVAD implantation and stay longer on intensive care unit than patients in group C (17.61 ± 16.96 vs 9.56 ± 9.42; P = .047). After exclusion, INTERMACS 1 patients it was not significant (14.8 ± 10.8 vs 9.8 ± 9.6 days; P = .065), the 30- and 90-day mortality was comparable. CONCLUSIONS: Implantation of pulsatile POLVAD MEV and continuous flow devices as LVAD support provides comparable results. A greater number of complications in group P can cause increased mortality over a longer observation period.

Effects of Left Ventricular Assist Device Support on End-Organ Function in Patients With Heart Failure: Comparison of Pulsatile- and Continuous-Flow Support in a Single-Center Experience.

BACKGROUND: Limited data exist about the effects of continuous-flow versus pulsatile-flow left ventricular assist devices (LVADs) on end-organ function. We hypothesized that a pulsatile Polvad MEV (PM) would result in outcomes similar to those of similarly ill patients implanted with a continuous-flow LVAD (Heartware [HW] or Heartmate II [HMII]). We aimed to compare renal, hepatic, and hematologic functions in the 1st 30 days of support. METHODS: We retrospectively reviewed patients with 24 PM (21 M, 3 F; group P) and 15 HW and 5 HMII (20 M, 0 F); group C LVAD implantations from April 2007 to February 2014. Creatinine, bilirubin, aspartate (AST) and alanine (ALT) transaminases, hematocrit, platelet count, international normalized ratio (INR), and activated partial thromboplastin time (APTT) parameters were analyzed before implantation and during 30 days of support. Demographic parameters were similar. RESULTS: No significant differences were found between the groups regarding baseline renal, hepatic, or hematologic function. Baseline INR and APTT were significantly higher in group P. Levels of creatinine were similar between groups. They increased from baseline to postoperative day (POD) 1 and then decreased. Bilirubin levels were insignificantly higher in group P. Transaminases were significantly higher in group P (AST in PODs 3-6, ALT in PODs 3-7). INR values were significantly higher at baseline and in POD 0. APTT values were insignificantly higher in group P. CONCLUSIONS: The use of LVAD improved renal and hepatic function in our series. Patients in group P had more decreased hepatic function and presented slower regeneration.

Right Ventricular Function in Patients With Left Ventricular Assist Device Support by Pulsatile Polvad MEV and Continuous-Flow Pumps Heartware and Heartmate II.

BACKGROUND: Left ventricular assist device (LVAD) support is increasingly used in patients with heart failure. The right ventricle (RV) plays a main role in LVAD support. Little is known about the effects of pulsatile Polvad MEV devices or continuouseconds flow pumps on RV function. We compared hemodynamic parameters of RV in patients after implantation of Polvad MEV (PM) and Heartware (HW) or Heartmate II (HMII) LVADs. METHODS: Forty-four patients were retrospectively reviewed after implantation of PM (group P; n = 24 [21 M, 3 F]) or HW or HMII (group C; n = 20 [20 M, 0 F]) LVADs from April 2007 to February 2014. Hemodynamic data-mean pulmonary pressure (mPAP), central venous pressure (CVP), cardiac output (CO), and cardiac index (CI)-were collected before surgery, after surgery, and every 2 hours in the intensive care unit, with the time points numbered from 1 to 120. Right ventricular work (RVW) was calculated according to the equation: RCW = CO × (mPAP - CVP) × 0.0144 (g·m). RESULTS: Baseline characteristic of the patients were similar. mPAP values were similar between groups. CVP values were higher in group P, significantly at time points 5 and 7-33. CO values were higher in group C, significantly from point 3 and almost all the time to point 43. CI reached significance at point 9, 12-14, 16-19, and 30-41. RCW was higher in group P before implantation. Post-implantation RCW values were higher in group C, significantly at time points 19, 20, 32-34, 51-53, and 55-57. CONCLUSIONS: Continuous-flow pumps more effectively optimize RV function than pulsatile LVADs, which can result in more effective prevention of RV failure or insuffiency in that group.

Contrasting denaturation maps of Xenopus laevis and Xenopus borealis mitochondrial DNAs.

Denaturation maps of mitochondrial DNAs of Xenopus laevis and Xenopus borealis are radically different from each other. This is in striking contrast to the invariant denaturation patterns previously recognized among mtDNAs of various Drosophila species, particularly, since the two toads may be even more closely related to each other than the Drosophila species.

Effects of the bradykinin B1 receptor antagonist des-Arg9[Leu8]bradykinin and genetic disruption of the B2 receptor on nociception in rats and mice.

The contributions of B1 and B2 bradykinin receptors to acute and chronic inflammatory hyperalgesia were examined using the peptide B1 receptor antagonist des-Arg9[Leu8]bradykinin and transgenic Bk2r-/- mice. In normal rats and mice, des-Arg9[Leu8]bradykinin (30 nmol/kg i.v. or s.c.) inhibited carrageenan-induced hyperalgesia and the late phase nociceptive response to formalin. The active dose range was narrow, suggesting partial agonist activity of this peptide. In rats with monoarthritis, des-Arg9[Leu8]bradykinin (up to 30 nmol/kg i.v.) failed to reduce the number of vocalisations elicited by gentle flexion and extension of the inflamed limb; however, hyperalgesia was exacerbated by administration of the B1 receptor agonist des-[Arg9]bradykinin (100 nmol/kg i.v.), consistent with other evidence for local induction of B1 receptors during adjuvant-induced arthritis. The nociceptive response to intraplantar injection of bradykinin (10 nmol) and hyperalgesia induced by carrageenan (0.6 mg) were absent in Bk2r-/- mice, indicating that stimulation of B2 receptors is an essential step in the initiation of some nociceptive and inflammatory reactions. However, the nociceptive response to formalin (2.5% intraplantar), including inhibition of the late phase by des-Arg9[Leu8]bradykinin (0.3 nmol), and induction of thermal hyperalgesia by Freund's adjuvant (0.1%) appeared intact in Bk2r-/- mice. These findings support other evidence for an involvement of B1 receptors in inflammatory hyperalgesia and suggest that B1 receptor antagonists may be clinically useful as anti-inflammatory and analgesic drugs.

Expression of B1 and B2 bradykinin receptor mRNA and their functional roles in sympathetic ganglia and sensory dorsal root ganglia neurones from wild-type and B2 receptor knockout mice.

Bradykinin has been implicated in nociception and inflammation. To examine the relative significance of B1 and B2 bradykinin receptor subtypes in sympathetic and sensory ganglia, the electrophysiological effects of bradykinin analogues and the expression of receptor subtype mRNA were examined in wild-type and "B2 knockout" mice from which the B2 receptor gene had been deleted. In wild-type mice the B2 receptor agonist bradykinin depolarized superior cervical ganglia (SCG) and activated inward currents in dorsal root ganglia (DRG) neurones. Responses to the B1 receptor agonist, [des-Arg10]-kallidin, were seen only in SCG that had been pre-treated with interleukins and the peptidase inhibitor captopril, but not in DRG neurones. The up-regulation of responses to [des-Arg10]-kallidin and substance P were blocked by indomethacin and, thus, were dependent upon cyclo-oxygenase activity. The effects of bradykinin were abolished in SCG and DRG's from B2 knockout mice and this was correlated with the absence of B2 receptor mRNA in ganglia from these animals. However, despite the presence of B1 receptor mRNA in interleukin treated SCG from B2 knockout mice, no depolarizing effects of the B1 receptor agonist [des-Arg10]-kallidin were observed. The successful elimination of bradykinin responses and B2 mRNA in sympathetic and sensory ganglia from B2 knockout mice, confirms that B2 receptors are the predominant functional bradykinin receptor subtype in these tissues and that B1 receptor mRNA is expressed in both sympathetic and sensory ganglia from these animals.

Experience with and comparision of methods of reduction mammaplasty.

Experinece with bilateral reduction mammaplasty in 213 patients has been reviewed. Complications, while not infrequent, are rarely of a serious or lasting nature. In the experience of the authors, the McKissock procedure has yielded the most consistently good results, although very good results were occasionally achieved with all of the other techniques employed.

The memory-facilitating effects of the competitive NMDA-receptor antagonist CGP 37849 are steroid-sensitive, whereas its memory-impairing effects are not.

The retention performance of mice in a passive-avoidance task was facilitated by low doses (0.3 mg/kg) of the competitive NMDA-receptor blocker CGP 37849, but impaired by high doses (30 mg/kg). The facilitatory effect was selectively suppressed by elevation of the plasma levels of aldosterone or corticosterone, or by blockade of steroid biosynthesis or the mineralocorticoid receptors. The impairment of memory, on the other hand, was not steroid sensitive. Accordingly, the data are in line with the hypothesis that drug induced memory facilitation is dependent on steroid sensitive processes.

Pretreatment with aldosterone or corticosterone blocks the memory-enhancing effects of nimodipine, captopril, CGP 37,849, and strychnine in mice.

Oral pretreatment with aldosterone or corticosterone blocked the memory-enhancing effects of the calcium antagonist nimodipine, the ACE inhibitor captopril, the NMDA blocker CGP 37,849, and the glycine antagonist strychnine in a passive-avoidance test in mice. The memory-disturbing effects of phenobarbitone, diazepam, CGP 37,849 and scopolamine were not influenced by the hormonal pretreatment. These findings could indicate the involvement of a steroid-sensitive mechanism in drug-induced improvement of memory. In the light of clinical observations showing elevated cortisol levels in Alzheimer patients, the results might also explain why only a limited number of these patients respond to therapy with memory enhancers.

The memory-enhancing effects of the piracetam-like nootropics are dependent on experimental parameters.

The effects of the nootropic agent piracetam and its congeners oxiracetam, pramiracetam and aniracetam on the retention performance of mice in a passive-avoidance situation are dependent on the intensity of the foot-shock applied. This phenomenon is observed upon both pre-trial and post-trial drug administration.

The GABAB receptor antagonist CGP 36,742 and the nootropic oxiracetam facilitate the formation of long-term memory.

The memory-enhancing effects of a single treatment with the GABAB antagonist CGP 36,742 (10 mg/kg) or the nootropic agent oxiracetam (100 mg/kg) given immediately after a learning experience ('post-trial') remain detectable for at least 4 months thereafter. This indicates that in all probability these substances facilitate the formation of the long-term memory trace.

Relation between age and clearance rate of nine investigational anticancer drugs from phase I pharmacokinetic data.

Aging influences the disposition and effects of several classes of drugs. Although drug clearance rate is correlated with toxicity for many anticancer drugs, few data have been published concerning the relationship of aging and clearance of chemotherapy. This study was performed to identify any relationship between age and clearance rate for anticancer drugs in phase I trials at the Johns Hopkins Oncology Center. In a retrospective study, we examined the clinical and pharmacokinetic data for 344 adults (aged 21-77 years) who received 9 phase I drugs with linear clearance in 13 clinical trials. We sought correlations between age and clearance for each drug and for the whole group. Data available for 9 of the 13 trials were used to compare age (< 65 or > 65 years) versus dose delivered [< the maximum tolerated dose (MTD) vs > or = the MTD] or toxicity (< grade 3 vs > or = grade 3). Of 344 patients, 81 (23.5%) were > 65 years old, 34 (9.9%) were > or = 70 years old, and 5 (1.5%) were > or = 75 years old. There was no significant correlation between drug clearance and age for individual drugs or the group as a whole. There was no significant difference between patients of the older and younger age groups with regard to dose or toxicity. Although only a small number of patients aged > or = 75 years were treated, our results suggest that the elderly do not experience greater toxicity even when treated at doses comparable with those given younger patients and should not be excluded from phase I trials on the basis of age. As the population of the United States ages, more elderly patients will be candidates for chemotherapy. A more thorough examination of the relationships between age, clearance rate, and toxicity can be accomplished as active drugs enter phase II/III studies.

How long does 'memory consolidation' take? New compounds can improve retention performance, even if administered up to 24 hours after the learning experience.

The 'nootropics' are a new class of psychoactive substances that improve learning and memory. Their almost exclusive effect on memory may indicate that they act on processes specifically involved in information storage. When administered after the learning trial, these substances improve subsequent retention performance in mice, even if an interval of 8 h has elapsed between learning and treatment. CGS 5649B, a highly active new substance, is effective even after an interval of 24 h. Although consonant with the 'consolidation' hypothesis, the results may challenge prevailing notions about the formation of memory traces.

Involvement of a steroidal component in the mechanism of action of piracetam-like nootropics.

Since adrenalectomy abolishes the memory-enhancing effects of piracetam and its derivatives, oxiracetam, aniracetam and pramiracetam, the question arises whether endogenous steroids play a role in their mechanism of action. We show that inhibition of steroid biosynthesis by aminoglutethimide and blockade of the aldosterone receptors by epoxymexrenone completely suppress the memory-improving effects of the nootropics. These results indicate that steroids, or, more precisely, activities mediated by the aldosterone receptors, might be involved in the mechanism of action of this class of nootropics. Blockade of aldosterone receptors, however, does not block the effects of cholinomimetics on memory, indicating the involvement of another mechanism of action.

Soluble selectin profiles associated with severe trauma.

Severe trauma acts as a trigger for the complex cascade of postinjury events leading to the release of different mediators and the development of generalized inflammation. Selectins are a family of adhesion proteins that are responsible for the adherence of polymorphonuclear neutrophils to the endothelium. This interaction plays an important role in the development of severe complications after multiple trauma. The aim of the present study is to follow the sequential alterations in circulating selectin levels after severe injury and to evaluate the clinical significance of these mediators in monitoring prognosis and outcome. Thirty four severely traumatized patients were entered into the study. Serum sE-selectin, plasma sP-selectin and sL-selectin concentrations were measured and an APACHE II score was calculated on admission to the intensive care unit and during the subsequent 5 days. The patients were divided into survivors and nonsurvivors. Initial soluble P- and E-selectin concentrations were significantly elevated in all trauma patients. The highest values of these adhesion molecules were measured in all the observed days in patients with poor prognosis and outcome. In survivors we found a systematic decrease in the sP-selectin concentrations. On admission, the sL-selectin concentrations in all trauma patients were decreased. There were stable, very low values in nonsurvivors and a slow increase in circulating L-selectin in patients who survived. The pattern of soluble selectins in patients with severe trauma is characterized by increased levels of P- and E-selectin and a decreased concentration of L-selectin. These findings suggest a widespread microvascular endothelial activation on injury in the early posttraumatic period, which may be associated with increased neutrophil-endothelial adhesion, neutrophil extravasation and migration. We suppose that these parameters of endothelial cell activation/injury may be useful as another early prognostic factor in severe trauma.