Study of a mechanism responsible for potential antidepressant activity of EMD 386088, a 5-HT6 partial agonist in rats.
It was shown that 5-HT6 receptor agonists can exert pharmacological activity due to various modifications in monoamines' level and metabolism activity in rats' brain structures. This finding was correlated with antidepressant- or anxiolytic-like properties of these compounds. The study was designed to establish a possible mechanism of the antidepressant-like activity of the partial 5-HT6 receptor agonist EMD386088 (5-chloro-2-methyl-3-(1,2,3,6-tetrahydro-4-pyridinyl)-1H-indole hydrochloride) in rats. The concentrations of monoamines (dopamine (DA), noradrenaline (NA), and serotonin (5-HT)) and the rate of their metabolism were measured ex vivo in the brain structures (hippocampus, nucleus accumbens, striatum) using high-performance liquid chromatography (HPLC). The rats were killed after the forced swim test (FST); the collected tissue samples were used to ex vivo experiments. The potency of EMD386088 to blockade dopamine transporter (DAT) was tested in a functional in vitro study. FST was used to assess the involvement of D1- and D2-like receptor subfamilies in antidepressant-like properties of EMD386088. Neurochemical data from ex vivo experiments showed that antiimmobility activity of EMD386088 may be connected with the activation of dopaminergic system, while neither noradrenergic nor serotonergic ones are involved in its effect. EMD386088 also possesses a significant affinity for DAT which may be a mechanism in the abovementioned effect. Behavioral data seem to confirm the importance of dopaminergic system activation in antidepressant-like activity of EMD386088, since this effect, observed in the FST, was abolished by the preferential D1- and D2-like receptor subfamily antagonists SCH23390 and sulpiride, respectively. Dopaminergic system is involved in antidepressant-like activity of EMD386088.


Optimization and characterization of a novel internally-cooled radiofrequency ablation system with optimized pulsing algorithm in an ex-vivo bovine liver.
Purpose: To prospectively characterize and optimize radiofrequency energy deposition to determine ideal parameters for achieving large ablation zones. Materials and methods: An internally-cooled RF system was used to perform 214 ablations in 72 ex-vivo bovine livers. Tip exposure (1-5 cm), electrode current (400-2500 mA), and application duration (3-15 min) were systematically varied. A pulsing algorithm optimized efficiency of RF deposition, including initial automatic ramping followed by adjustment in current, in response to changes in tissue impedance. Following the procedure ablation diameter and length were measured, sphericity calculated, and correlated with parameters of energy deposition and tissue temperatures. Results: Increasing electrode exposure from 1-5 cm produced linear increases in ablation diameter from 1.4 ± 0.1 to 5.3 ± 0.1 cm (y = 1.1x-0.5; R2 = 0.93), and length (y = 1.18x + 0.34; R2 = 0.92). A sphericity index >0.85 was noted at optimal energy setting for electrode exposures of 1-4 cm. Maximum temperatures post-ablation increased with active tip length from 68.5 ± 4.9 °C to 91.3 ± 1.5 °C in a logarithmic (y = 0.94ln(x)-2.75; R2 = 0.90) or power relationship between temperature and the resultant ablation diameter (y = 0.27e0.0295x; R2 = 0.76). A tight exponential relationship (y = 0.28x0.38; R2 = 0.97) was also observed between total energy deposition and ablation diameter. Finally, a multifactor relationship of the diameter of ablation to electrode tip exposure and the time to first impedance rise was successfully modeled, with a root mean squared error of 1.9 mm and R2 = 0.95. Conclusion: Large, reproducible, and spherical ablation areas can be achieved with the novel system described, with efficient delivery of RF energy deposited into tissue. These findings may have important clinical relevance in regards to the clinical utility of RF ablation compared to other competitive forms of thermal tumor ablation.


Nanoscale imaging of electronic surface transport probed by atom movements induced by scanning tunneling microscope current.
The hopping movements of Cl atoms on a Si(111)-(7 x 7) surface that are enhanced by an electron injection from tips of a scanning tunneling microscope (STM) exhibit a spatial spread from the electron injection point with an anisotropic distribution. The enhanced hopping effect becomes greatest at a sample bias voltage being resonant with the Si-Cl antibonding states and also exhibits an oscillatory decay with the distance from the injection point characterized by the wavelength depending on the bias voltage. All of these facts can be interpreted in terms of the coherent expansion of the electron wave packets locally formed at the STM tip.


Effects of pH on Nanofibrillation of TEMPO-Oxidized Paper Mulberry Bast Fibers.
TEMPO oxidation was conducted as a pretreatment to achieve efficient nanofibrillation of long paper mulberry bast fibers (PMBFs). The pH dependency of nanofibrillation efficiency and the characteristics of the resulting cellulose nanofibrils (CNFs) were investigated. As the pH increased, the negative value of the zeta potential of TEMPO-oxidized fibers increased. The increase in electrostatic repulsion at pH values of greater than 9 prevented the entanglement of long PMBFs, which was a drawback for defibrillation at acidic pH. With increasing pH, the CNF production yield was increased. The crystallinity index of TEMPO-oxidized CNFs from PMBFs was 83.5%, which was higher than that of TEMPO-oxidized CNFs from softwood fibers in the same conditions. The tensile strength of nanopaper from TEMPO-oxidized PMBF CNFs was 110.18 MPa, which was approximately 30% higher than that (84.19 MPa) of the TEMPO-oxidized CNFs from softwood fibers.


Acute pulmonary edema associated with placement of waist-high, custom-fit compression stockings.
Compression stockings are a safe, noninvasive treatment for patients with symptomatic orthostatic hypotension due to autonomic nervous system dysfunction. In this report, we describe a 75-year-old man who had development of pulmonary edema approximately 45 minutes after placement of compression stockings on the first postoperative day following a carotid endarterectomy. No sudden changes were noted on an electrocardiogram or echocardiogram or in the cardiac isoenzymes associated with the pulmonary edema. The patient had a history of coronary artery disease, diabetes mellitus-induced autonomic nervous system dysfunction, and recent surgery near the carotid baroreceptor. All these factors may have limited his ability to compensate for a rapid increase in central blood volume. The temporal relationship of the patient's respiratory distress to the placement of the compression stockings, in the absence of laboratory findings of primary cardiac dysfunction, make stocking-related fluid shift the likely precipitating event in the formation of acute pulmonary edema. This case suggests that compression stockings should be used with caution in patients with limited cardiac reserve.


Cortical spreading depression induces propagating activation of the thalamus ventral posteromedial nucleus in awake mice.
As the relay centre for processing sensory information, the thalamus may involve in the abnormal sensory procedure caused by cortical spreading depression (CSD). However, few studies have focused on the transient response of thalamus during CSD. Our study aimed to investigate the neuronal activity of mouse thalamus ventral posteromedial nucleus (VPM) during CSD by in vivo micro-endoscopic fluorescence imaging of the genetic calcium probe GCaMP6s expressed in excitatory glutamatergic neurons. Thirty-four transgenic VGluT2-GCaMP6s mice were used in the experiments. An endoscope was inserted into the VPM for image acquisition. CSD was induced by KCl topically applied unilaterally on the cranial dura. Data were acquired in awake (ipsilateral or contralateral VPM, saline instead of KCl, MK-801 treatment) and anaesthetized (isoflurane, pentobarbital) states. Statistical analysis was performed using analysis of variance (ANOVA) by SPSS. We found that after CSD induced in ipsilateral motor cortex, the neuronal activity increased and propagated from the posterior-lateral to the anterior-medial part of the VPM with an average speed of 3.47 mm/min. When CSD was induced in visual cortex, the response propagated in opposite direction, from the anterior-medial to the posterior-lateral part of the VPM. Aanaesthetics resulted in the suppression of VPM activation induced by CSD. No significant VPM activation was detected when CSD was induced in contralateral cortex or KCl was replaced by saline. When 5 mM MK-801 was applied to the dura, the electrode failed to record the DC shift of CSD, and there was no significant VPM activation after KCl application. CSD induced propagating activation of the ipsilateral VPM in awake mice. The response might correlate to the cortical location where CSD was induced and might be affected by anaesthetics. No significant VPM activation was detected in saline and mk801 experiment results indicated that this VPM activation is due to CSD rather than mouse motion or direct effect of the KCl applying to the intact dura. This finding suggests the potential involvement of thalamus in the migraine auras.


Peritoneal cytology and its prognostic value in endometrial carcinoma.
There has been a controversy about the prognostic significance of positive peritoneal cytology in endometrial carcinoma. Peritoneal cytology was obtained at the time of surgery, including systematic retroperitoneal lymph node dissection, in 114 patients. The incidence of positive peritoneal cytology was 35.1%. The 5-year survival rates of the stage IIIA and IIIC (FIGO, 1988) cases were 82.8% and 58.3%, respectively. In pathological stage I (the disease was histologically confined to the uterine corpus), there was no significant difference in 5-year survival rates between patients with and without positive peritoneal cytology. Though the patients in stage IIIA who had only positive peritoneal cytology were given no postoperative therapy unless they had extrauterine disease, no patients developed recurrence. In stages IIIC and IV, the prognosis was significantly poorer for patients with positive peritoneal cytology than for those with negative cytology. Positive peritoneal cytology is not an adverse prognostic factor endometrial carcinoma if disease is limited to the uterus.


Molecular characterization of feline type I interferon receptor 2.
The cDNA sequence of feline interferon receptor 2 (feIFNAR2) was generated using RT-PCR method in present study. This gene included 1,572 bp and encoded a 523 aminoacid (aa) protein with a 35 aa signal peptide. The deduced protein shared 61% amino acid identity to the human IFNAR2. There were two fibronectin type III (FBN-III) domains of about 110 residues in the extracellular domain. Homology modeling of feIFNAR2 presented a similar structure with other IFN receptors. The ELISA and FACS experiments demonstrated that the protein could bind to feIFN-alpha or feIFN-omega. However, antiviral activity assay found that feIFN-omega had broader species spectrum compared with feIFN-alpha. To define the functional differences, several point mutations of feIFNAR2 were constructed and the relative affinities of feIFN-alpha or feIFN-omega for feIFNAR2 and mutants were evaluated. The results suggested that feIFN-alpha and feIFN-omega had different binding sites on feIFNAR2. T75 and M77 on feIFNAR2 were hotspots for binding to feIFN-alpha, but not to feIFN-omega. These findings suggested that the cloned feline IFNAR2 interacted with both feIFN-alpha and feIFN-omega, however, not sharing the same binding sites.


The Predictive Value of Stair Climbing Test on Postoperative Complications in Lung Cancer Patients with Limited Pulmonary Function.
To evaluate the predictive value of stair climbing test (SCT) on postoperative complications in lung cancer patients with limited pulmonary function. A total of 727 hospitalized lung cancer patients with limited pulmonary function were retrospectively reviewed. Included in the cohort were 424 patients who underwent SCT preoperatively. Patients were grouped according to general condition, past medical history, surgical approach, pulmonary function test, and SCT results. Comparison of the postoperative cardiopulmonary complication rates was made and independent risk factors were identified. A total of 89 cardiopulmonary-related complications occurred in 69 cases, accounting for 16.3% of the entire cohort. The postoperative cardiopulmonary complication rates were significantly different between groups stratified by smoking index, percentage of forced expiratory volume in one second, percentage of diffusion capacity for carbon monoxide, SCT results, excision extension, and anesthetic duration (p <0.05). Multivariate analysis showed that only height achieved (p <0.001), changes in heart rate (∆HR; p <0.001), and excision extension (p = 0.006) were independent risk factors for postoperative cardiopulmonary complications. The SCT could be used as a preoperative screening method for lung cancer patients with limited pulmonary function. For those patients who could only climb less than 6 floors or had ∆HR >30 bpm in the test, sublobar resection should be selected to reduce the postoperative cardiopulmonary complication rate.


Can we know what Terri Schiavo would have wanted?
What would Terri Schiavo have wanted? That remains an unanswered question for many who followed the media frenzy that attended the extraordinary court and legislative battles that preceded her death 13 days after her feeding tube was removed for the last time. What would she have directed her physicians to do if she had "miraculously" regained capacity and awareness of the consequences of her cardiac arrest that left her in a persistent vegetative state? Who would she have wanted to make that decision for her if she were unable to do so? How are we to understand the meaning of statements that she purportedly made about life-sustaining treatments approximately 20 years ago, and how can we apply them to the current situation? This article reflects on those questions from the perspective of two small exploratory studies. These studies considered the meanings and interpretation of statements by terminally ill patients concerning desire for hastened death and the relevance of previously made statements to their current clinical situation.


Topical 5-fluorouracil has no additional benefit in treating common warts with cryotherapy: a single-centre, double-blind, randomized, placebo-controlled trial.
The role of topical 5-fluorouracil (5-FU) in treating common warts is not well defined. We tried to evaluate the efficacy and adverse effects of combination cryotherapy and topical 5% 5-FU ointment in the treatment of common warts. The study was a single-centre, double-blind randomized placebo-controlled trial. In the study, 80 patients with common warts were randomized into two groups and underwent two 10-second freeze/thaw cycles of cryotherapy with liquid nitrogen once every three weeks for a maximum of five treatments. Between treatments, patients applied either topical 5% 5-FU ointment (group A) or placebo aqueous cream (group B) twice daily. The mean +/- SD reduction in wart area was 58.57 +/- 0.06% in group A and 65.29 +/- 0.06% in group B. In total, 19 patients in group A and 24 patients in group B had wart size reduced by 75% or more (P = 0.50), while 12 patients in group A and 17 patients in group B had clearance of their warts (P = 0.245). Logistic regression with age, sex, smoking status, immune status, site, duration and number of warty lesions, history of previous treatment, and treatment group found that only a history of previous treatment and acral lesions were significant adverse predictors of improvement. There was no significant difference in the number of adverse events between the two groups, although there was a trend towards more pain and blistering associated with topical 5-FU. We concluded that topical 5-FU has no added benefit in treating common warts with cryotherapy.


The perioperative morbidity of scheduled cesarean hysterectomy.
To compare the outcomes of patients undergoing scheduled cesarean hysterectomy with those of women treated with cesarean delivery and subsequent hysterectomy. Through a retrospective review of 43 patients, we investigated the morbidity associated with scheduled cesarean hysterectomy and compared these findings with the combined morbidity of scheduled cesarean delivery and subsequent abdominal hysterectomy in a control population. Controls were included only if the subsequent hysterectomy was performed within 3 years of the index cesarean delivery. Each study subject was assigned two controls matched for age, parity, number of previous cesarean deliveries, and indications for procedures. The incidence of the following major morbidity events was compared between the groups: transfusion, urinary tract injury, fistula formation, cellulitis or endometritis, postoperative abscess or hematoma formation, ileus, pneumonia, and wound complications requiring prolonged therapy (seroma, hematoma, infection). The number of women receiving transfusions after scheduled cesarean hysterectomy was greater than among controls (39.5 versus 15.1%; P < .05). The proportion of patients with major morbidity, exclusive of transfusion, was significantly greater in the control population (44%) than in women with scheduled cesarean hysterectomy (16%) (P < .05). The cumulative number of women with a major complication, such as transfusion or a morbid event, was 22 of 43 in the study group versus 44 of 86 in the control population, a nonsignificant difference. We found no significant difference in the cumulative perioperative complication rates in women undergoing scheduled cesarean hysterectomy compared with a population of similar patients treated with cesarean delivery and subsequent abdominal hysterectomy.


A3243G mitochondrial DNA mutation in Tunisian diabetic population.
An excess of maternal transmission of adult onset diabetes mellitus has been observed in the studied Tunisian patients, in fact, diabetic patients with affected mother are significantly more important than those with affected father (p< 10-6) There is increasing evidence that mtDNA mutations may be involved in this disease, since mitochondrial transmission offers a plausible explanation for a proportion of this maternal excess comparing to paternal transmission. The aim of the present study was to investigate the mitochondrial DNA involvement in the inheritance of diabetes in Tunisian population and to evaluate the frequency of substitution A3243G in these patients. In the current study we investigated for the first time, the 3243 mtDNA in 280 Tunisian diabetic patients. Results showed that the frequency of this substitution in tRNALeu is about 1.07%. This percentage is similar to those reported in Japanese, German and French populations.


The impact of a multidisciplinary approach on caring for ventilator-dependent patients.
To determine the clinical and financial outcomes of a highly structured multidisciplinary care model for patients in an intensive care unit (ICU) who require prolonged mechanical ventilation. The structured model outcomes (protocol group) are compared with the preprotocol outcomes. Descriptive study with financial analysis. A twelve-bed medical-surgical ICU in a non-teaching tertiary referral center in Ogden, Utah. During a 54 month period, 469 consecutive intensive care patients requiring mechanical ventilation for longer than 72 hours who did not meet exclusion criteria were studied. A multidisciplinary team was formed to coordinate the care of ventilator-dependent patients. Care was integrated by daily collaborative bedside rounds, monthly meetings, and implementation of numerous guidelines and protocols. Patients were followed from the time of ICU admission until the day of hospital discharge. Patients were assigned APACHE II scores on admission to the ICU, and were divided into eight diagnostic categories. ICU length of stay, hospital length of stay, costs, charges, reimbursement, and in-hospital mortality were measured. Mortality in the preprotocol and protocol group, after adjustment for APACHE II scores, remained statistically unchanged (21-23%). After we implemented the new care model, we demonstrated significant decreases in the mean survivor's ICU length of stay (19.8 days to 14.7 days, P= 0.001), hospital length of stay (34.6 days to 25.9 days, P=0.001), charges (US$102500 to US$78500, P=0.001), and costs (US$71900 to US$58000, P=0.001). Implementation of a structured multidisciplinary care model to care for a heterogeneous population of ventilator-dependent ICU patients was associated with significant reductions in ICU and hospital lengths of stay, charges, and costs. Mortality rates were unaffected.


Mammalian lignan formation in rats fed a wheat bran diet.
The dietary origin of lignan phytoestrogens is still poorly understood more than 20 years after their discovery in human urine. Their level in urine has been associated with the consumption of dietary fiber. This paper reports the study of the excretion of enterolactone, assayed by a time-resolved fluoroimmunoassay, in rats fed a diet supplemented with 15% wheat bran, one of the main sources of fiber in Western countries. Enterolactone excretion regularly increased during the two weeks of the diet to reach a value of 45 nmol/day. The level of excretion also increased upon preadaptation to ferulic acid, structurally related to secoisolariciresinol, an established precursor of enterolactone in flaxseeds, and decreased upon preadaptation to potato starch rich in fiber. These results show that the formation of lignans from wheat bran is influenced by the diet, possibly because of an adaptation of the colonic microflora.


Examination of the ability of the nasal administration route to confer a brain exposure advantage for three chemical inhibitors of P-glycoprotein.
The central nervous system (CNS), efficiently isolated from the systemic circulation by the blood-brain barrier (BBB), represents a challenging therapeutic target. For CNS-targeted agents, augmenting brain exposure by increasing blood drug concentrations often is prohibited by systemic toxicity. Therefore, a means for selectively increasing brain exposure, while minimizing systemic exposure, would be desirable. Limited evidence has indicated that nasally-administered compounds can penetrate into brain, although the selectivity of this approach is unclear. This study demonstrated a distinct, but compound-specific, advantage of the nasal administration route in conferring selective CNS delivery (defined as a brain exposure advantage; BEA). Brain and systemic concentrations of three P-glycoprotein-inhibiting agents were evaluated following single nasal or systemic doses to mice, and the influence of administration route on brain exposure (absolute BEA) and on brain-to-blood partitioning (relative BEA) was calculated. Relative and absolute BEA differed markedly among rifampin, quinidine, and GF120918, with relative BEA ranging between 1.53- and 809-fold and absolute BEA between 0.114- and 9.19-fold. Although substantial increases in brain exposure and partitioning in conjunction with nasal administration were demonstrated, the utility of this approach may be limited by inability to deliver a therapeutically relevant mass of drug to the brain.


Criteria for entrapped gas under a drop on an ultrahydrophobic surface.
Ultrahydrophobicity of a rough surface is mainly attributed to the entrapped gas under a drop. Two criteria were proposed for the entrapped gas: an intruding angle criterion and an intruding depth criterion. These two criteria are that the intruding angle must be less than the maximum asperity slope angle and the intruding depth must be less than the height of the asperities. The intruding angle is determined by the true contact angle, the surface geometry, and the drop size. The intruding angle is directly proportional to the true contact angle, and it increases with an increase of the fractional area of the liquid-gas interface under the drop and with a decrease of the linear dimension of the three-phase contact line on the asperities. The effect of the drop size on the intruding angle is induced by Laplace and hydrostatic pressures. The intruding depth increases with an increase of the intruding angle and the distance between the asperities. The proposed criteria were evaluated using experiment data from the literature. Comparison between the experiment and calculation results showed that the experiment data supported the theory.


The number and localisation of adenine nucleotide-binding sites in beef-heart mitochondrial ATPase (F1) determined by photolabelling with 8-azido-ATP and 8-azido-ADP.
1. When irradiated 8-azido-ATP becomes covalently bound (as the nitreno compound) to beef-heart mitochondrial ATPase (F1) as the triphosphate, either in the absence or presence of Mg2+, label covalently bound is not hydrolysed. 2. In the presence of Mg2+ the nitreno-ATP is bound to both the alpha and beta subunits, mainly (63%) to the alpha subunits. 3. After successive photolabelling of F1 with 8-azido-ATP (no Mg2+) and 8-azido-ADP (with Mg2+) 4 mol label is bound to F1, 2 mol to the alpha and 2 mol to the beta subunits. 4. When the order of photolabelling is reversed, much less 8-nitreno-ATP is bound to F1 previously labelled with 8-nitreno-ADP. It is concluded that binding to the alpha-subunits hinders binding to the beta subunits. 5. F1 that has been photolabelled with up to 4 mol label still contains 2 mol firmly bound adenine nucleotides per mol F1. 6. It is concluded that at least 6 sites for adenine nucleotides are present in isolated F1.


Empowering health care improvement: an operational model.
The shift from compliance to continuous improvement requires a bridge--empowerment--that enables the creation of an environment in which employees can contribute the full range of their expertise in service to the organization. Leaders are responsible for creating this empowering environment. An operational model that describes the nine aspects of empowerment (for example, mission and values, access to information, avenues of influence) can serve as a template for leaders to use to promote an atmosphere of empowerment in their organizations. The authors adapted this model as a survey instrument to uncover some hospital success stories that demonstrate several aspects of empowerment in action. The survey was administered at two hospitals. CASE STUDY 1: At Beth Israel Hospital (Boston, Mass), Nutrition Services rated the "mission and vision" aspect of empowerment in their department at an average of 3.5 out of a possible 4.0. The department attributes this high rating to their focus on specific goals that reinforce the larger mission and vision of the hospital as a whole. These goals form the basis for annual employee evaluations. For example, a 1992 patient-focused goal was to provide nutrition counseling and education. The department approached this goal by developing a daily format ("Eat Smart") for the daily hospital cafeteria menu that provided nutritional information. The rationale was to educate employees, who could then bring back information to their patients. Alignment of the department's efforts with the larger mission of the hospital enabled Nutrition Services to extend its influence beyond departmental and professional boundaries. Implementation of other plans at the hospital demonstrate a sustained process of vision development on the part of the executive leadership. CASE STUDY 2: At Robert Wood Johnson University Hospital (New Brunswick, NJ), Support Services gave the "environment of trust and respect" aspect an average rating of 3.7. The key to this success story lies in the upgraded role of the housekeepers as "hosts." The "Pro-Act" restructuring model for patient care delivery creates and upgrades positions to relieve nurses of their task loads. The three nursing services that participated in the survey gave their highest rating to the "access to the education and training" aspect of empowerment. The high quality of professional excellence is supported by tuition reimbursement, incentives for credentials, the extern program and scholarships provided to attract outstanding nursing students to their staff and encouragement of nursing staff at all levels for publication, presentations, and poster sessions. These case studies demonstrate how leaders created an environment in which collective learning has become a way of doing business. Given the multifaceted nature of the empowerment model, some may wonder where to begin to focus leadership efforts. It is important to emphasize that the constellation of empowerment aspects constitutes an integral system; the full potential for empowerment arises from synergy among all nine aspects.


The effect on protein and amino acid metabolism of an intravenous nutrition regimen providing seventy percent of nonprotein calories as lipid.
The purpose of this study was to determine which peripheral intravenous nutrition (IVN) regimen, one containing 70% nonprotein calories as lipid IVN or one containing all nonprotein calories as glucose IVN, was most effective at reversing some effects of surgery on protein metabolism. Twenty patients who required IVN after operation were randomized into two well-matched groups that received 36 kcal.kg-1.day-1 glucose IVN or 37 kcal.kg-1.day-1 lipid IVN. Both IVN regimens resulted in similar changes of nitrogen balance, plasma liver enzymes, blood urea, plasma albumin, and plasma prealbumin. Mean plasma transferrin rose significantly after glucose IVN (p less than 0.01), a change greater than that after lipid IVN (p less than 0.05). Lipid IVN resulted in continued net efflux of alanine from peripheral tissues at rates similar to pretreatment values; glucose IVN significantly reduced net alanine efflux (p less than 0.05). Most of the extra alanine produced by peripheral tissues during lipid IVN appeared to derive from an increased uptake of intramuscular glutamate and from an increased uptake of branched chain amino acids. The results of this study indicate that 38 kcal.kg-1.day-1 lipid IVN was equivalent to glucose IVN, except for the continuing gluconeogenesis of alanine and the delayed recovery of plasma transferrin concentration after surgery. A greater infusion rate of such a regimen may be necessary to provide sufficient glucose to suppress gluconeogenesis.


Trocar site closure: a new and easy technique.
The GORE (W.L. Gore & Associates, Phoenix, AZ) Suture Passer, a new reusable fascial closure device, is one of the best devices on the market for closure of laparoscopic trocar site wounds. The technique for closure of fascia using this device is described. It is a cost-effective, safe, and easy.


Perioperative dilutional coagulopathy treated with fresh frozen plasma and fibrinogen concentrate: a prospective randomized intervention trial.
Treatment of dilutional coagulopathy by transfusing fresh frozen plasma (FFP) remains sub-optimal. We hypothesized that partial replacement of transfused FFP by fibrinogen concentrate results in improved coagulant activity and haemostasis. This was tested in a controlled clinical intervention trial with patients experiencing massive bleeding during major surgery. Patients undergoing major elective surgery were treated according to current protocols. When transfusion with FFP was required, patients were randomized as follows: group A received 4 units FFP and group B received 2 units FFP plus 2 g fibrinogen concentrate. Blood samples were taken before and after the intervention. Analysts were blinded to the treatment type. Group A (B) consisted of 21 (22) patients, in 16 (17) of whom bleeding stopped after intervention. Plasma fibrinogen increased significantly more in group B (0·57 g/l) than in group A (0·05 g/l). However, levels of prothrombin and factors VIII, IX and X increased more in group A than in group B. Rotational thromboelastometry (ROTEM) of whole blood and plasma revealed improved fibrin clot formation in group B but not in group A. Thrombin generation [calibrated automated thrombogram (CAT)] in plasma increased more in group A. Principal parameters determining whole-blood thromboelastometry were the fibrinogen level and platelet count. In vitro addition of fibrinogen and prothrombin complex concentrate to pre-intervention samples restored both ROTEM and CAT parameters. Partial replacement of transfused FFP by fibrinogen increases fibrin clot formation at the expense of less improved thrombin generation. Coagulation factors other than fibrinogen alone are required for full restoration of haemostasis.


Do children need concurrent prompts in order to use lexical analogies in reading?
In two experiments, children were taught a word and then given reminders of that word while reading unfamiliar analogous words. Reminders of the pronunciation of taught words facilitated reading of rime analogous words (e.g. beak-peak) over head analogous words (e.g. beak-bean) in Experiment 1, and onset analogous words (e.g. stilt-stem), over medial analogous words (e.g. stilt-milk) in Experiment 2. Importantly, however, privileged improvement for rime and onset analogous words was not evident in the absence of these concurrent prompts. The findings suggest that the analogy model developed by Goswami (1993) may have limited applicability in naturalistic settings.


Reintegration of Brewers Spent Grains in the Food Chain: Nutritional, Functional and Sensorial Aspects.
Nowadays, the pandemic situation has encouraged the idea of sustainable healthy foods leading to new trends in food consumption. Brewers spent grain (BSG) represents a potential functional food rich in fiber, protein, lipids, mineral and phenols that needs to be further exploited. In this vein, five different BSG types were collected from local breweries and valorized in cookies manufacturing. Thus, proximate composition (protein, minerals, lipids, ash, crude fiber and carbohydrates) was analyzed using AACC (American Association of Cereal Chemists) methods, DPPH (2,2-Diphenyl-1-picrylhydrazyl), and Folin Ciocalteu methods were used to determined antioxidant activity and total phenols, while minerals and aroma volatile compounds were performed using inductively coupled plasma optical emission spectrometry (ICP-OES) and ITEX/GC-MS (in tube extraction gas chromatography-mass spectrometry) respectively. Color and physical characteristics, together with sensorial analysis, were also evaluated. The results highlighted a significant difference between BSG samples, mainly from the total phenols, antioxidant activity and aroma volatile compounds point of view. BSG volatiles compounds from the aldehydes group such as 2-methyl-propanal, 3-methyl-butanal and 2-methyl-butanal were identified also in the final baked goods, leading to a pleasant and appreciated consumers' taste and aroma. Furthermore, cookies sensorial analysis emphasized that the sample manufactured with BSG from light and dark malt mixture was more appreciated by consumers, attaining the highest hedonic scores.


Contact Forces between Single Metal Oxide Nanoparticles in Gas-Phase Applications and Processes.
In this work we present a comprehensive experimental study to determine the contact forces between individual metal oxide nanoparticles in the gas-phase using atomic force microscopy. In addition, we determined the amount of physisorbed water for each type of particle surface. By comparing our results with mathematical models of the interaction forces, we could demonstrate that classical continuum models of van der Waals and capillary forces alone cannot sufficiently describe the experimental findings. Rather, the discrete nature of the molecules has to be considered, which leads to ordering at the interface and the occurrence of solvation forces. We demonstrate that inclusion of solvation forces in the model leads to quantitative agreement with experimental data and that tuning of the molecular order by addition of isopropanol vapor allows us to control the interaction forces between the nanoparticles.


Anatomic distribution of the growth-associated protein GAP-43 in the developing human brainstem.
GAP-43 is a membrane phosphoprotein whose expression is high in neurons undergoing development or remodeling of axonal connections. This study used a monospecific antibody to GAP-43 to investigate the sequences of fiber tract elongation and synaptic development in the human brainstem. Immunocytochemistry was performed in 14 fetal and infant brainstems; two child and adult cases were also examined for comparison. At midgestation, GAP-43 immunostaining was moderately intense across nuclei and fiber tracts, except for the corticospinal tract, where levels were higher, and cranial nerve nucleus VII, superior olive, inferior colliculus, inferior olivary hilum, inferior cerebellar peduncle, medial lemniscus, and medial longitudinal fasciculus, where staining was nearly absent. By the end of the neonatal period, the relative distribution of GAP-43 immunostaining appeared well-established and similar, although not identical, to that in the child and adult brainstem. Immunostaining was absent or negligible in almost all the cranial nerve somato- and branchiomotor nuclei, auditory-relay nuclei, and vestibular nuclei, while remaining intense in visceral-related nuclei, reticular formation, cochlear nucleus, and periaqueductal gray. Staining was also virtually absent in all fiber tracts at birth, except for the corticospinal tract and central tegmental tract. Persistence of GAP-43 staining in the corticospinal tract past the fetal period suggests that this tract remains in a plastic state beyond initial axonal elongation. Intense immunostaining in visceral-related nuclei into adulthood suggests that these regions may continue to undergo synaptic reorganization. This study provides baseline information relevant to understanding developmental brainstem disorders in early human life.


The value of 5-aminosalicylic acid in inflammatory bowel disease for patients intolerant or allergic to sulphasalazine.
The use of sulphasalazine in inflammatory bowel disease is often limited by intolerance or allergy. In previous studies 5-aminosalicylic acid (5ASA), the active ingredient of sulphasalazine, coated with an acrylic resin, has been shown to be as effective as sulphasalazine in maintaining remission. In this study coated 5ASA was given to 37 patients intolerant or allergic to sulphasalazine. Thirty-three patients have tolerated 5ASA satisfactorily, and most experienced improved control of their disease. Four patients reported similar side effects with sulphasalazine and 5ASA.


Identifying the active site in nitrogen-doped graphene for the VO2+/VO2(+) redox reaction.
Nitrogen-doped graphene sheets (NGS), synthesized by annealing graphite oxide (GO) with urea at 700-1050 °C, were studied as positive electrodes in a vanadium redox flow battery. The NGS, in particular annealed at 900 °C, exhibited excellent catalytic performance in terms of electron transfer (ET) resistance (4.74 ± 0.51 and 7.27 ± 0.42 Ω for the anodic process and cathodic process, respectively) and reversibility (ΔE = 100 mV, Ipa/Ipc = 1.38 at a scan rate of 50 mV s(-1)). Detailed research confirms that not the nitrogen doping level but the nitrogen type in the graphene sheets determines the catalytic activity. Among four types of nitrogen species doped into the graphene lattice including pyridinic-N, pyrrolic-N, quaternary nitrogen, and oxidic-N, quaternary nitrogen is verified as a catalytic active center for the [VO](2+)/[VO2](+) couple reaction. A mechanism is proposed to explain the electrocatalytic performance of NGS for the [VO](2+)/[VO2](+) couple reaction. The possible formation of a N-V transitional bonding state, which facilitates the ET between the outer electrode and reactant ions, is a key step for its high catalytic activity.


[Molecular markers for the blood coagulation and fibrinolysis].
It is now possible to more precisely evaluate coagulation and fibrinolysis by laboratory tests in which various molecular markers can be measured. The determination of the plasma level of soluble fibrin may be the most important molecular marker for the diagnosis of thrombosis and disseminated intravascular coagulation. Combined with the results of the other molecular marker determinations, the measurement of the plasma level of soluble fibrin might provide useful information about the coagulation status of patients.


Thermal motions and function of bacteriorhodopsin in purple membranes: effects of temperature and hydration studied by neutron scattering.
The internal dynamics of bacteriorhodopsin, the light-driven proton pump in the purple membrane of Halobacterium halobium, has been studied by inelastic neutron scattering for various conditions of temperature and hydration. Light activation can take place when the membrane is vibrating harmonically. The ability of the protein to functionally relax and complete the photocycle initiated by the absorption of a photon, however, is strongly correlated with the onset of low-frequency, large-amplitude anharmonic atomic motions in the membrane. For a normally hydrated sample, this occurs at about 230 K, where a dynamical transition from a low-temperature harmonic regime is observed. In moderately dry samples, on the other hand, in which the photocycle is slowed down by several orders of magnitude, no transition is observed and protein motions remain approximately harmonic up to room temperature. These results support the hypothesis, made from previous neutron diffraction studies, that the "softness" of the membrane modulates the function of bacteriorhodopsin by allowing or not allowing large-amplitude motions in the protein.


The epigenetic clock as a predictor of disease and mortality risk: a systematic review and meta-analysis.
Ageing is one of the principal risk factors for many chronic diseases. However, there is considerable between-person variation in the rate of ageing and individual differences in their susceptibility to disease and death. Epigenetic mechanisms may play a role in human ageing, and DNA methylation age biomarkers may be good predictors of age-related diseases and mortality risk. The aims of this systematic review were to identify and synthesise the evidence for an association between peripherally measured DNA methylation age and longevity, age-related disease, and mortality risk. A systematic search was conducted in line with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Using relevant search terms, MEDLINE, Embase, Cochrane Central Register of Controlled Trials, and PsychINFO databases were searched to identify articles meeting the inclusion criteria. Studies were assessed for bias using Joanna Briggs Institute critical appraisal checklists. Data was extracted from studies measuring age acceleration as a predictor of age-related diseases, mortality or longevity, and the findings for similar outcomes compared. Using Review Manager 5.3 software, two meta-analyses (one per epigenetic clock) were conducted on studies measuring all-cause mortality. Twenty-three relevant articles were identified, including a total of 41,607 participants. Four studies focused on ageing and longevity, 11 on age-related disease (cancer, cardiovascular disease, and dementia), and 11 on mortality. There was some, although inconsistent, evidence for an association between increased DNA methylation age and risk of disease. Meta-analyses indicated that each 5-year increase in DNA methylation age was associated an 8 to 15% increased risk of mortality. Due to the small number of studies and heterogeneity in study design and outcomes, the association between DNA methylation age and age-related disease and longevity is inconclusive. Increased epigenetic age was associated with mortality risk, but positive publication bias needs to be considered. Further research is needed to determine the extent to which DNA methylation age can be used as a clinical biomarker.


Salt an Essential Nutrient: Advances in Understanding Salt Taste Detection Using Drosophila as a Model System.
Taste modalities are conserved in insects and mammals. Sweet gustatory signals evoke attractive behaviors while bitter gustatory information drive aversive behaviors. Salt (NaCl) is an essential nutrient required for various physiological processes, including electrolyte homeostasis, neuronal activity, nutrient absorption, and muscle contraction. Not only mammals, even in Drosophila melanogaster, the detection of NaCl induces two different behaviors: Low concentrations of NaCl act as an attractant, whereas high concentrations act as repellant. The fruit fly is an excellent model system for studying the underlying mechanisms of salt taste due to its relatively simple neuroanatomical organization of the brain and peripheral taste system, the availability of powerful genetic tools and transgenic strains. In this review, we have revisited the literature and the information provided by various laboratories using invertebrate model system Drosophila that has helped us to understand NaCl salt taste so far. We hope that this compiled information from Drosophila will be of general significance and interest for forthcoming studies of the structure, function, and behavioral role of NaCl-sensitive (low and high concentrations) gustatory circuitry for understanding NaCl salt taste in all animals.


The Range and Scientific Value of Randomized Trials.
The randomized, controlled trial (RCT) is the gold standard of scientific evidence for the attribution of clinical effects (benefits and harms) to medical interventions. Many different designs for RCTs have been developed in order to counter legitimate critical objections and to better adapt the trials to the continually changing challenges that face clinical research. The diversity and adaptability of randomized trial designs are presented and discussed on the basis of a selective literature review and specific illustrative examples. A wide range of RCT designs enables adaptation to special research tasks and clinical framework conditions. These include (among others) crossover trials, n=1 trials, factorial RCT designs, and cluster-randomized trials. In addition, adaptive designs such as modern platform trials and pragmatic RCTs with simplified clinical questions and less severely restricted patient groups make broad recruitment of patients possible even in routine clinical practice. Only the randomized allocation of subjects to the treatment and control groups, which is the defining property of RCTs, can adequately ensure that traits of the subjects which might disturb or bias a comparison of two or more medical interventions, will be evenly distributed across groups, regardless of whether these traits are known or unknown. The methodological variants and further elaborations of the RCT that are discussed here will help protect patients by enabling the assessment of the benefits and harms of medical methods and products on the basis of robust evidence even in the present era of rapid innovation.


Determinants of COVID-19 vaccine acceptance in the US.
The COVID-19 pandemic continues to adversely affect the U.S., which leads globally in total cases and deaths. As COVID-19 vaccines are under development, public health officials and policymakers need to create strategic vaccine-acceptance messaging to effectively control the pandemic and prevent thousands of additional deaths. Using an online platform, we surveyed the U.S. adult population in May 2020 to understand risk perceptions about the COVID-19 pandemic, acceptance of a COVID-19 vaccine, and trust in sources of information. These factors were compared across basic demographics. Of the 672 participants surveyed, 450 (67%) said they would accept a COVID-19 vaccine if it is recommended for them. Males (72%) compared to females, older adults (≥55 years; 78%) compared to younger adults, Asians (81%) compared to other racial and ethnic groups, and college and/or graduate degree holders (75%) compared to people with less than a college degree were more likely to accept the vaccine. When comparing reported influenza vaccine uptake to reported acceptance of the COVID-19 vaccine: 1) participants who did not complete high school had a very low influenza vaccine uptake (10%), while 60% of the same group said they would accept the COVID-19 vaccine; 2) unemployed participants reported lower influenza uptake and lower COVID-19 vaccine acceptance when compared to those employed or retired; and, 3) Black Americans reported lower influenza vaccine uptake and lower COVID-19 vaccine acceptance than all other racial groups reported in our study. Lastly, we identified geographic differences with Department of Health and Human Services (DHHS) regions 2 (New York) and 5 (Chicago) reporting less than 50 percent COVID-19 vaccine acceptance. Although our study found a 67% acceptance of a COVID-19 vaccine, there were noticeable demographic and geographical disparities in vaccine acceptance. Before a COVID-19 vaccine is introduced to the U.S., public health officials and policymakers must prioritize effective COVID-19 vaccine-acceptance messaging for all Americans, especially those who are most vulnerable.


A Critical Function for the Transcription Factors GLI1 and GLI2 in the Proliferation and Survival of Human Mast Cells.
Mast cell hyperactivity and accumulation in tissues are associated with allergy and other mast cell-related disorders. However, the molecular pathways regulating mast cell survival in homeostasis and disease are not completely understood. As glioma-associated oncogene (GLI) proteins are involved in both tissue homeostasis and in the hematopoietic system by regulating cell fate decisions, we sought to investigate the role for GLI proteins in the control of proliferation and survival of human mast cells. GLI1 transcripts were present in primary human mast cells and mast cell lines harboring or not activating mutations in the tyrosine kinase receptor KIT (HMC-1.1 and HMC-1.2, and LAD2 cells, respectively), while GLI2 transcripts were only present in HMC-1.1 and HMC-1.2 cells, suggesting a role for oncogenic KIT signaling in the regulation of GLI2. Reduction in GLI activity by small molecule inhibitors, or by shRNA-mediated knockdown of GLI1 or GLI2, led to increases in apoptotic cell death in both cultured human and murine mast cells, and reduced the number of peritoneal mast cells in mice. Although GLI proteins are typically activated via the hedgehog pathway, steady-state activation of GLI in mast cells occurred primarily via non-canonical pathways. Apoptosis induced by GLI silencing was associated with a downregulation in the expression of KIT and of genes that influence p53 stability and function including USP48, which promotes p53 degradation; and iASPP, which inhibits p53-induced transcription, thus leading to the induction of p53-regulated apoptotic genes. Furthermore, we found that GLI silencing inhibited the proliferation of neoplastic mast cell lines, an effect that was more pronounced in rapidly growing cells. Our findings support the conclusion that GLI1/2 transcription factors are critical regulators of mast cell survival and that their inhibition leads to a significant reduction in the number of mast cells in vitro and in vivo, even in cells with constitutively active KIT variants. This knowledge can potentially be applicable to reducing mast cell burden in mast cell-related diseases.


Studies on eye irritation caused by chemicals in rabbits--II. Structure-activity relationships and in vitro approach to primary eye irritation of salicylates in rabbits.
Structure-activity relationships and in vitro evaluation of eye irritation potential of salicylates in rabbits were studied. The primary eye irritation potential of ten salicylates was evaluated according to Draize method. The effects of chemicals on model protein and lipid were investigated in vitro. The effects of chemicals on the protein could be detected by the production of aggregates of human serum gamma-globulin (HSG) and a good correlation was obtained between the ability of salicylates to produce aggregation of HSG and the potential of corneal irritation. The effects on the lipid could be detected by the adhesion potential of chemicals on lipid membrane and a linear correlation was not obtained between the adhesionary effects of salicylates on lipid membrane and the potential eye irritation. The corneal irritation and protein aggregation potential of salicylates were correlated with the acid dissociation constant more closely than octanol/water partition coefficient. The destruction of alpha-helix of proteins in corneal surface by salicylates were observed from the nondestructive structural analysis of corneal surface by Fourier Transform (FT)-IR spectroscopy. These results suggest that eye irritation caused by salicylates are mainly the results of denaturation of proteins in ocular tissue and that the effects on protein depend on the dissociation potential of molecules.


No expression of human leukocyte antigen G (HLA-G) in colorectal cancer cells.
Although there is a specific antitumor immune response in the body, colorectal cancer cells progressively develop. This fact indicated that the cancer cells could have a variety of mechanisms to evade or escape the immune system. HLA-G is identified to inhibit the recognition of NK-cell in various kinds of cancers. This study investigated the expression of HLA-G in colorectal cancer. Eighty five specimens of colorectal cancer, carcinoma in situ and adenomatous polyp were examined by immunohistochemistry and RT-PCR for the detection of human leukocyte antigen (HLA)-G The expression of HLA-G was not found in all colorectal specimens (85/85) both protein level and transcription level, suggesting that the expression of HLA-G is not a possible immune evasion mechanism of colorectal cancer cell.


Total parenteral nutrition and tumor metastasis.
A major clinical concern with providing total parenteral nutrition to the tumor-bearing host is the potential to stimulate tumor growth. This study was performed to determine the effect of total parenteral nutrition on primary tumor growth and spontaneous pulmonary metastasis in Lobund rats with subcutaneous prostate adenocarcinoma (PA-III) implants. Significant acceleration of primary tumor growth and tumor metastasis occurred in animals receiving parenteral nutrition consisting of amino acids, dextrose, and lipid or standard enteral nutrition compared to control animals. This study represents the first report of stimulating tumor metastasis with specific parenteral nutrients and clearly indicates that both primary tumor growth and tumor metastasis can be altered by exogenous substrate administration.


QSAR Analysis of a Series of Hydantoin-based Androgen Receptor Modulators and Corresponding Binding Affinities.
Androgen receptor (AR), a member of the nuclear hormone receptor superfamily of intracellular ligand-dependent transcription factors, plays an indispensable role in normal male development through the regulation of androgen through the binding with endogenous androgens. Inappropriate amounts of androgens have a severe adverse effect on men. Excessive androgen may contribute to accelerate prostatic hypertrophy, even prostate cancer, while the absence of androgen may result in reduced muscle mass and strength, decreased bone mass, low energy, diminished sexual function and an increased risk of osteoporosis and fracture. In these cases, androgen receptor modulators are important to maintain the normal biological function of AR. So androgen receptor modulators are necessary for human being to improve their happy life index. To explore the relationships between molecular structures and corresponding binding abilities to aid the new AR modulator design, multiple linear regressions (MLR) are employed to analyze a series of hydantoin analogues, which can bind to androgen receptor acting as AR modulators. The obtained optimum model presents wonderful reliabilities and strong predictive abilities with R2 =0.858, QLOO2 =0.822, QLMO2 =0.813, QF12 =0.840, QF22 =0.807, QF32 =0.814, CCC=0.893, respectively. The derived model can be used to predict the binding abilities of unknown chemicals and may help to design novel molecules with better AR affinity activity.


Pathogen virulence genes--implications for vaccines and drug therapy.
The emergence and spread of bacteria resistant to antimicrobial drugs is a major public health problem with a growing number of infections becoming virtually untreatable. There is a need to develop interventions both to prevent and to treat diseases caused by multi-resistant microbes. We review some recently developed methods (including whole genome nucleotide sequencing projects) to study bacterial pathogenesis, and discuss how knowledge gained from understanding the molecular mechanisms of disease can be applied to combat the threat of infectious diseases.


Diverse clinical presentations of celiac disease in the same family.
We performed a family study to evaluate a total of 34 extended family members (8 siblings, 23 children and nephews, and 3 grandchildren) of an adult patient with celiac disease (CD), a 58- year-old male with severe neurologic involvement manifested as myoclonias. We found 3 other members affected with CD (a 44-year old sister, a 39-year old niece, and a 26-year old nephew). Two of them were completely asymptomatic and all had hypertransaminasemia. All exhibited a villous atrophy pattern of the duodenal mucosa (1 mild, 1 moderate, 1 severe). Overall family involvement was 11.8% (4/14). We wish to emphasize the need to perform extended family studies when diagnosing a case of CD, since risk is not restricted to only first-degree relatives.


DHEA protects against visceral obesity and muscle insulin resistance in rats fed a high-fat diet.
Visceral obesity is frequently associated with muscle insulin resistance. Rats fed a high-fat diet rapidly develop obesity and insulin resistance. Dehydroepiandrosterone (DHEA) has been reported to protect against the development of obesity. This study tested the hypothesis that DHEA protects against the increase in visceral fat and the development of muscle insulin resistance induced by a high-fat diet in rats. Feeding rats a diet providing 50% of the energy as fat for 4 wk resulted in a twofold greater visceral fat mass and a 50% lower rate of maximally insulin-stimulated muscle 2-deoxyglucose (2-DG) uptake compared with controls. Rats fed the high-fat diet plus 0.3% DHEA were largely protected against the increase in visceral fat (+ 11.3 g in high fat vs. + 2.9 g in high fat plus DHEA, compared with controls) and against the decrease in insulin-stimulated muscle 2-DG uptake (0.94 +/- 0.15 mumol.ml-1.20 min-1, controls; 0.46 +/- 0.06 mumol.ml-1.20 min-1, high-fat diet; 0.78 +/- 0.07 mumol.ml-1.20 min-1, high fat + DHEA). DHEA did not affect food intake. These results show that DHEA has a protective effect against accumulation of visceral fat and development of muscle insulin resistance in rats fed a high-fat diet.


The role of hydrogels with tethered acetylcholine functionality on the adhesion and viability of hippocampal neurons and glial cells.
In neural tissue engineering, designing materials with the right chemical cues is crucial in providing a permissive microenvironment to encourage and guide neuronal cell attachment and differentiation. Modifying synthetic hydrogels with biologically active molecules has become an increasingly important route in this field to provide a successful biomaterial and cell interaction. This study presents a strategy of using the monomer 2-methacryloxyethyl trimethylammonium chloride (MAETAC) to provide tethered neurotransmitter acetylcholine-like functionality with a complete 2-acetoxy-N,N,N-trimethylethanaminium segment, thereby modifying the properties of commonly used, non-adhesive PEG-based hydrogels. The effect of the functional monomer concentration on the physical properties of the hydrogels was systematically studied, and the resulting hydrogels were also evaluated for mice hippocampal neural cell attachment and growth. Results from this study showed that MAETAC in the hydrogels promotes neuronal cell attachment and differentiation in a concentration-dependent manner, different proportions of MAETAC monomer in the reaction mixture produce hydrogels with different porous structures, swollen states, and mechanical strengths. Growth of mice hippocampal cells cultured on the hydrogels showed differences in number, length of processes and exhibited different survival rates. Our results indicate that chemical composition of the biomaterials is a key factor in neural cell attachment and growth, and integration of the appropriate amount of tethered neurotransmitter functionalities can be a simple and effective way to optimize existing biomaterials for neuronal tissue engineering applications.


Development of an ostomy competency.
Staff educators and staff nurses developed an ostomy competency, with the guidance and expertise of the advanced practitioner and enterostomal nurse at a large teaching hospital. The competency improved the quality of care for surgical ostomy patients. Care was standardized and staff nurses' clinical knowledge was enhanced. Following the sessions, staff nurses verbalized increased confidence in working with patients with ostomies and demonstrated increased autonomy and problem-solving abilities. No variances in educational aspects of care were noted on clinical pathways.


Neuropsychological, Neurocognitive, Vestibular, and Neuroimaging Correlates of Exposure to Repetitive Low-Level Blast Waves: Evidence From Four Nonoverlapping Samples of Canadian Breachers.
We assessed the utility of a battery of neuropsychological, neurocognitive, physiological (balance, ataxia, postural tremor), and neuroimaging measures for studying the effects of blast waves in breachers-a population repeatedly exposed to low-level blast during military training and operations. Data were collected from four nonoverlapping samples, in the course of similarly structured 4-day breacher training exercises in successive years involving a combination of indoor and outdoor blast events. In all cases, self-report and neuropsychological measures were administered once at baseline (i.e., 1 day before the start of training). In years 1-2, neurocognitive and physiological measures were administered daily before and after training. In years 3-4, neurocognitive data were collected once at baseline. In Year 4, we introduced 3 modifications to our design. First, in addition to breachers, we also collected data from sex-and age-matched military controls at the same time points. Second, we assessed balance, ataxia, and postural tremor immediately following blast exposure "in the field," enabling us to quantify its acute effects. Third, structural magnetic resonance imaging (MRI) scans were acquired before and after the 4-day training exercise to explore differences between breachers and controls at baseline, as well as possible training-related changes using voxel-based morphometry. These design modifications were made to enable us to test additional hypotheses in the context of the same training exercise. At baseline, scores on the "Rivermead Post Concussion Symptoms Questionnaire," "RAND SF-36" (physical functioning, role limitation due to physical health, social functioning, energy/fatigue, general health), and "Short Musculoskeletal Function Questionnaire" distinguished breachers from controls. Also at baseline, the MRI data revealed that there was greater regional gray matter volume in controls compared to breachers in the right superior frontal gyrus. Balance, ataxia, and postural tremor did not exhibit sensitivity to the acute effects of blast in the field, nor did neurocognitive measures to its cumulative or daily effects. Our exploratory results suggest that self-report neuropsychological measures and structural MRI hold promise as sensitive measures for quantifying the long-term, cumulative effects of blast exposure in breachers. We discuss the limitations of our study and the need for prospective longitudinal data for drawing causal inferences regarding the impact of blast exposure on breachers' health and performance.


Communication and Cooperation Between the Medical Academy, Medical Association, and Local Government: Health Counseling Program After Recovery From Coronavirus Disease 2019 (COVID-19) in Daegu.
We are currently experiencing the disaster of the COVID-19 pandemic. Since the first case of Coronavirus disease 2019 (COVID-19) was confirmed in South Korea on January 20, the number of COVID-19 cases in South Korea has been rapidly increasing until early March due to a local spread in Daegu, which is one of the eight metropolitan cities in South Korea with a population of 2.5 million. As the medical academy has social accountability as professionals, Daegu-Gyeongbuk branch of the Korean Academy of Family Medicine (Daegu-Gyeongbuk branch) developed the health counseling program for discharged COVID-19 patients. The Daegu-Gyeongbuk branch communicated with Daegu Medical Association and Daegu city for this program and incorporated available resources and capabilities as a leader of this program. This newly developed counseling program consists of medical consultations, sending healthcare brochures and medical supplies, and the appraisal at the end of the program. Not only COVID-19 related symptoms but also other psychological problems are also dealt with during consultations. This program started on March 18, and over 1,700 recovered patients have been receiving counseling as of April 28. Communication and cooperation between the medical academy, medical association, and government are essential to overcome the COVID-19 pandemic. Besides, we expect to apply this health counseling program and our model of setting this program cooperating with medical association and government to different infectious pandemic crisis.


Angle-independent motion measurement by correlation of ultrasound signals assessed with a single circular-shaped transducer.
In medicine, pulsed ultrasound is a widespread noninvasive technique that measures motion in the direction of the ultrasound beam, i.e., axial motion. The magnitude of the actual motion can be determined only if the angle between the ultrasound beam and the direction of motion (transducer-to-motion angle) is known. For blood flow measurements, current pulsed ultrasound systems assume this angle to be equal to the angle between the ultrasound beam and the longitudinal direction of the vessel, as can be estimated from a two-dimensional brightness-mode (B-mode) image that is obtained prior to the blood flow measurement. For tissue motion measurements, current pulsed ultrasound systems are mostly unable to determine the transducer-to-motion angle. Recently, a model has been derived for the correlation of(analytic) radiofrequency (rf) signals, assessed with a circular-shaped ultrasound transducer along the same line of observation. In the present paper, this model is used to derive estimators, requiring only the calculation of a few correlation coefficients, for the motion components (axial, lateral and actual) and for some of the signal parameters (center frequency, bandwidth and signal-to-noise ratio) of the assessed rf signals. The transducer-to-motion angle can be derived from the estimated motion components. For the evaluation of the estimators, rf signals were acquired with a motion-controlled experimental arrangement. The results of the evaluation study show that the transducer-to-motion angle can be estimated with a mean standard deviation of less than 2 degrees.


Non-enzymatic protein modification by the Maillard reaction reduces the activities of scavenging enzymes in Vigna radiata.
The non-enzymatic modification of proteins through the Maillard reaction plays an important role in the loss of seed viability during seed storage. In the present study we examined whether the Maillard reaction reduces the activities of scavenging enzymes in Vigna radiata (mung bean) seeds during storage. Seeds were stored under various conditions for different duration. Maillard products were monitored by measuring protein fluorescence, and the activities of glutathione reductase (GR), superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and peroxidase (POX) were determined. The accumulation of Maillard products in seed axes increased during storage with increasing moisture content and temperature, and was correlated with the decline in seed vigour. The activities of GR, CAT and APX decreased in proportion to the increase in Maillard products at all the moisture contents and temperatures tested. These enzymatic changes were also correlated with seed vigour. However, the activities of SOD and POX remained unchanged and appeared to be less sensitive to the Maillard reaction.


Developmental and environmental signals induce distinct histone acetylation profiles on distal and proximal promoter elements of the C4-Pepc gene in maize.
The maize C(4)-Pepc gene is expressed in an organ- and cell-type-specific manner, inducible by light and modulated by nutrient availability and the metabolic state of the cell. We studied the contribution of histone acetylation at five lysine residues to the integration of these signals into a graduated promoter response. In roots and coleoptiles, where the gene is constitutively inactive, three of the five lysines were acetylated and the modifications showed unique patterns with respect to their distribution on the gene. A similar pattern was observed in etiolated leaves, where the gene is poised for activation by light. Here, illumination selectively induced the acetylation of histone H4 lysine 5 and histone H3 lysine 9 in both the promoter and the transcribed region, again with unique distribution patterns. Induction was independent of transcription and fully reversible in the dark. Nitrate and hexose availability modulated acetylation of all five lysines restricted to a distal promoter region, whereas proximal promoter acetylation was highly resistant to these stimuli. Our data suggest that light induction of acetylation is controlled by regulating HDAC activity, whereas metabolic signals regulate HAT activity. Acetylation turnover rates were high in the distal promoter and the transcribed regions, but low on the proximal promoter. On the basis of these results, we propose a model with three levels of stimulus-induced histone modifications that collectively adjust promoter activity. The results support a charge neutralization model for the distal promoter and a stimulus-mediated, but transcription-independent, histone acetylation pattern on the core promoter, which might be part of a more complex histone code.


Sulfation of Tyr1680 of human blood coagulation factor VIII is essential for the interaction of factor VIII with von Willebrand factor.
The acidic region of the Factor VIII light chain was studied with regard to structural requirements for the formation of a functional von Willebrand factor (vWF)-binding site. Factor VIII mutants lacking the B domain, with additional deletions and an amino acid replacement within the sequence 1649-1689 were constructed using site-directed mutagenesis and expressed in Cos-1 cells. These mutants, which were recovered as single-chain molecules with similar specific activities, were compared in their binding to immobilized vWF. Deletion of amino acids 741-1648 or 741-1668 did not affect the binding of Factor VIII to vWF. However, a mutant with a deletion of residues 741-1689 was no longer capable of interacting with vWF. This indicates a role for residues within the sequence 1669-1689 in the formation of a vWF-binding site. When recombinant Factor VIII was expressed in the presence of chlorate, an inhibitor of protein sulfation, the resulting Factor VIII displayed strongly reduced binding to vWF. vWF binding was completely abolished when within the sequence 1669-1689 the tyrosine residue Tyr1680, which is part of a consensus tyrosine sulfation sequence, was replaced by phenylalanine. The Factor VIII sequence 1673-1689 was identified as a high affinity substrate for tyrosylprotein sulfotransferase (Km = 57 microM) in cell-free sulfation studies. It is concluded that sulfation of Tyr1680 is required for the interaction of Factor VIII with vWF. Two synthetic peptides that represent the sequence 1673-1689, but differ with respect to sulfation of Tyr1680 are shown to have vWF binding affinity that is considerably lower than the Factor VIII protein. Several models to accommodate our findings are discussed.


Penetrating cations enhance uncoupling activity of anionic protonophores in mitochondria.
Protonophorous uncouplers causing a partial decrease in mitochondrial membrane potential are promising candidates for therapeutic applications. Here we showed that hydrophobic penetrating cations specifically targeted to mitochondria in a membrane potential-driven fashion increased proton-translocating activity of the anionic uncouplers 2,4-dinitrophenol (DNP) and carbonylcyanide-p-trifluorophenylhydrazone (FCCP). In planar bilayer lipid membranes (BLM) separating two compartments with different pH values, DNP-mediated diffusion potential of H(+) ions was enhanced in the presence of dodecyltriphenylphosphonium cation (C12TPP). The mitochondria-targeted penetrating cations strongly increased DNP- and carbonylcyanide m-chlorophenylhydrazone (CCCP)-mediated steady-state current through BLM when a transmembrane electrical potential difference was applied. Carboxyfluorescein efflux from liposomes initiated by the plastoquinone-containing penetrating cation SkQ1 was inhibited by both DNP and FCCP. Formation of complexes between the cation and CCCP was observed spectophotometrically. In contrast to the less hydrophobic tetraphenylphosphonium cation (TPP), SkQ1 and C12TPP promoted the uncoupling action of DNP and FCCP on isolated mitochondria. C12TPP and FCCP exhibited a synergistic effect decreasing the membrane potential of mitochondria in yeast cells. The stimulating action of penetrating cations on the protonophore-mediated uncoupling is assumed to be useful for medical applications of low (non-toxic) concentrations of protonophores.


Visualizing RAD51-mediated joint molecules: implications for recombination mechanism and the effect of sequence heterology.
The defining event in homologous recombination is the exchange of base-paired partners between a single-stranded (ss) DNA and a homologous duplex driven by recombinase proteins, such as human RAD51. To understand the mechanism of this essential genome maintenance event, we analyzed the structure of RAD51-DNA complexes representing strand exchange intermediates at nanometer resolution by scanning force microscopy. Joint molecules were formed between substrates with a defined ssDNA segment and homologous region on a double-stranded (ds) partner. We discovered and quantified several notable architectural features of RAD51 joint molecules. Each end of the RAD51-bound joints had a distinct structure. Using linear substrates, a 10-nt region of mispaired bases blocked extension of joint molecules in all examples observed, whereas 4 nt of heterology only partially blocked joint molecule extension. Joint molecules, including 10 nt of heterology, had paired DNA on either side of the heterologous substitution, indicating that pairing could initiate from the free 3'end of ssDNA or from a region adjacent to the ss-ds junction. RAD51 filaments covering joint ss-dsDNA regions were more stable to disassembly than filaments covering dsDNA. We discuss how distinct structural features of RAD51-bound DNA joints can play important roles as recognition sites for proteins that facilitate and control strand exchange.


Integrated analysis reveals that STAT3 is central to the crosstalk between HER/ErbB receptor signaling pathways in human mammary epithelial cells.
Human epidermal growth factor receptors (HER, also known as ErbB) drive cellular proliferation, pro-survival and stress responses by activating several downstream kinases, in particular ERK, p38 MAPK, JNK (SAPK), the PI3K/AKT, as well as various transcriptional regulators such as STAT3. When co-expressed, the first three members of HER family (HER1-3) can form homo- and hetero-dimers, and there is considerable evidence suggesting that the receptor dimers differentially activate intracellular signaling pathways. To better understand the interactions in this system, we pursued multi-factorial experiments where HER dimerization patterns and signaling pathways were rationally perturbed. We measured the activation of HER1-3 receptors and of the sentinel signaling proteins ERK, AKT, p38 MAPK, JNK, STAT3 as a function of time in a panel of human mammary epithelial (HME) cells expressing different levels of HER1-3 stimulated with various ligand combinations. We hypothesized that the HER dimerization pattern is a better predictor of downstream signaling than the total receptor activation levels. We validated this hypothesis using a combination of model-based analysis to quantify the HER dimerization patterns, and by clustering the activation data in multiple ways to confirm that the HER receptor dimer is a better predictor of the signaling through p38 MAPK, ERK and AKT pathways than the total HER receptor expression and activation levels. We then pursued combinatorial inhibition studies to identify the causal regulatory interactions between sentinel signaling proteins. Quantitative analysis of the collected data using the modular response analysis (MRA) and its Bayesian Variable Selection Algorithm (BVSA) version allowed us to obtain a consensus regulatory interaction model, which revealed that STAT3 occupies a central role in the crosstalk between the studied pathways in HME cells. Results of the BVSA/MRA and cluster analysis were in agreement with each other.


Diagnostic Benefit of High b-Value Computed Diffusion-Weighted Imaging in Patients with Hepatic Metastasis.
Diffusion-weighted imaging (DWI) has rapidly become an essential tool for the detection of malignant liver lesions. The aim of this study was to investigate the usefulness of high b-value computed DWI (c-DWI) in comparison to standard DWI in patients with hepatic metastases. In total, 92 patients with histopathologic confirmed primary tumors with hepatic metastasis were retrospectively analyzed by two readers. DWI was obtained with b-values of 50, 400 and 800 or 1000 s/mm2 on a 1.5 T magnetic resonance imaging (MRI) scanner. C-DWI was calculated with a monoexponential model with high b-values of 1000, 2000, 3000, 4000 and 5000 s/mm2. All c-DWI images with high b-values were compared to the acquired DWI sequence at a b-value of 800 or 1000 s/mm2 in terms of volume, lesion detectability and image quality. In the group of a b-value of 800 from a b-value of 2000 s/mm2, hepatic lesion sizes were significantly smaller than on acquired DWI (metastases lesion sizes b = 800 vs. b 2000 s/mm2: mean 25 cm3 (range 10-60 cm3) vs. mean 17.5 cm3 (range 5-35 cm3), p < 0.01). In the second group at a high b-value of 1500 s/mm2, liver metastases were larger than on c-DWI at higher b-values (b = 1500 vs. b 2000 s/mm2, mean 10 cm3 (range 4-24 cm3) vs. mean 9 cm3 (range 5-19 cm3), p < 0.01). In both groups, there was a clear reduction in lesion detectability at b = 2000 s/mm2, with hepatic metastases being less visible compared to c-DWI images at b = 1500 s/mm2 in at least 80% of all patients. Image quality dropped significantly starting from c-DWI at b = 3000 s/mm2. In both groups, almost all high b-values images at b = 4000 s/mm2 and 5000 s/mm2 were not diagnostic due to poor image quality. High c-DWI b-values up to b = 1500 s/mm2 offer comparable detectability for hepatic metastases compared to standard DWI. Higher b-value images over 2000 s/mm2 lead to a noticeable reduction in imaging quality, which could hamper diagnosis.


Effect of rest interval on neuromuscular and metabolic responses between children and adolescents.
The purpose of this study was to compare the effect of different rest intervals and contraction velocities on muscle recovery following resistance exercise. 18 children (11.1 ± 0.52 yrs) and 19 adolescents (15.8 ± 0.49 yrs) performed three sets of 10 isokinetic repetitions at 60°/s and 180°/s. The work-to-rest ratio (W/R) was 1:2 and 1:4 for 60°/s, and 1:6 and 1:12 for 180°/s. ANOVA revealed that children demonstrated no significant decline in PT from the first to third set with any rest interval, but there was a significant (p < .05) decline for adolescents when a W/R of 1:2, 1:4 and 1:6 were used. Adolescents demonstrated significantly greater blood lactate (BLa) concentrations than children after three sets of resistance exercise. The present study indicates that adolescents may require longer rest intervals to recover full PT when compared with children.


Age-dependent accumulation of oligomeric SNCA/α-synuclein from impaired degradation in mutant LRRK2 knockin mouse model of Parkinson disease: role for therapeutic activation of chaperone-mediated autophagy (CMA).
Parkinson disease (PD) is an age-related neurodegenerative disorder associated with misfolded SNCA/α-synuclein accumulation in brain. Impaired catabolism of SNCA potentiates formation of its toxic oligomers. LRRK2 (leucine-rich repeat kinase-2) mutations predispose to familial and sporadic PD. Mutant LRRK2 perturbs chaperone-mediated-autophagy (CMA) to degrade SNCA. We showed greater age-dependent accumulation of oligomeric SNCA in striatum and cortex of aged LRRK2R1441G knockin (KI) mice, compared to age-matched wildtype (WT) by 53% and 31%, respectively. Lysosomal clustering and accumulation of CMA-specific LAMP2A and HSPA8/HSC70 proteins were observed in aged mutant striatum along with increased GAPDH (CMA substrate) by immunohistochemistry of dorsal striatum and flow cytometry of ventral midbrain cells. Using our new reporter protein clearance assay, mutant mouse embryonic fibroblasts (MEFs) expressing either SNCA or CMA recognition 'KFERQ'-like motif conjugated with photoactivated-PAmCherry showed slower cellular clearance compared to WT by 28% and 34%, respectively. However, such difference was not observed after the 'KFERQ'-motif was mutated. LRRK2 mutant MEFs exhibited lower lysosomal degradation than WT indicating lysosomal dysfunction. LAMP2A-knockdown reduced total lysosomal activity and clearance of 'KFERQ'-substrate in WT but not in mutant MEFs, indicating impaired CMA in the latter. A CMA-specific activator, AR7, induced neuronal LAMP2A transcription and lysosomal activity in MEFs. AR7 also attenuated the progressive accumulation of both intracellular and extracellular SNCA oligomers in prolonged cultures of mutant cortical neurons (DIV21), indicating that oligomer accumulation can be suppressed by CMA activation. Activation of autophagic pathways to reduce aged-related accumulation of pathogenic SNCA oligomers is a viable disease-modifying therapeutic strategy for PD.Abbreviations: 3-MA: 3-methyladenine; AR7: 7-chloro-3-(4-methylphenyl)-2H-1,4-benzoxazine; CMA: chaperone-mediated autophagy; CQ: chloroquine; CSF: cerebrospinal fluid; DDM: n-dodecyl β-D-maltoside; DIV: days in vitro; ELISA: enzyme-linked immunosorbent assay; FACS: fluorescence-activated cell sorting; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GWAS: genome-wide association studies; HSPA8/HSC70: heat shock protein 8; KFERQ: CMA recognition pentapeptide; KI: knockin; LAMP1: lysosomal-associated membrane protein 1; LAMP2A: lysosomal-associated membrane protein 2A; LDH: lactate dehydrogenase; LRRK2: leucine-rich repeat kinase 2; MEF: mouse embryonic fibroblast; NDUFS4: NADH:ubiquinone oxidoreductase core subunit S4; NE: novel epitope; PD: Parkinson disease; RARA/RARα: retinoic acid receptor, alpha; SNCA: synuclein, alpha; TUBB3/TUJ1: tubulin, beta 3 class III; WT: wild-type.


Cancer of unknown primary site: a review of 28 cases and the efficacy of cisplatin/docetaxel therapy at a single institute in Japan.
We evaluated the efficacy and toxicity of cisplatin/docetaxel (CDDP/TXT) chemotherapy and identified prognostic factors in Japanese patients with cancer of unknown primary site (CUP). Twenty-eight consecutive patients seen at a single institute were reviewed retrospectively. Sixteen patients were treated with TXT 80 mg/m2, followed by CDDP 75 mg/m2. The overall response rate to CDDP/TXT treatment was 62.5%, with a median survival time (MST) of 22.7 months. Common adverse reactions were myelosuppression and hyponatremia. The MST of all 28 patients with CUP was 8.3 months, and the 1-year overall survival rate was 45.6%. Univariate analysis identified 5 prognostic factors:performance status, liver involvement, bone involvement, pleural involvement, and lymph node involvement. In conclusion, CDDP/TXT chemotherapy is effective with tolerable toxicity in patients with CUP. Japanese patients with CUP might be chemosensitive and may survive longer.


Activation of serotonin receptors promotes microglial injury-induced motility but attenuates phagocytic activity.
Microglia, the brain immune cell, express several neurotransmitter receptors which modulate microglial functions. In this project we studied the impact of serotonin receptor activation on distinct microglial properties as serotonin deficiency not only has been linked to a number of psychiatric disease like depression and anxiety but may also permeate from the periphery through blood-brain barrier openings seen in neurodegenerative disease. First, we tested the impact of serotonin on the microglial response to an insult caused by a laser lesion in the cortex of acute slices from Cx3Cr1-GFP-/+ mice. In the presence of serotonin the microglial processes moved more rapidly towards the laser lesion which is considered to be a chemotactic response to ATP. Similarly, the chemotactic response of cultured microglia to ATP was also enhanced by serotonin. Quantification of phagocytic activity by determining the uptake of microspheres showed that the amoeboid microglia in slices from early postnatal animals or microglia in culture respond to serotonin application with a decreased phagocytic activity whereas we could not detect any significant change in ramified microglia in situ. The presence of microglial serotonin receptors was confirmed by patch-clamp experiments in culture and amoeboid microglia and by qPCR analysis of RNA isolated from primary cultured and acutely isolated adult microglia. These data suggest that microglia express functional serotonin receptors linked to distinct microglial properties.


Species diversity and detection of pathogens in phlebotomine sand flies collected from forest management areas of Quintana Roo, Mexico.
Sand flies have expanded their areas of distribution, thereby increasing the risk of pathogen transmission in non-endemic areas. To establish efficient prevention and control strategies for the transmission of vector-borne pathogens, it is important to understand seasonal dynamics of their vectors. In Mexico, there are several areas where the contact between sand flies, hosts and reservoirs favours the transmission of the pathogen. We compared sand fly communities in a forest management area and a conserved area in Noh-Bec, Quintana Roo, Mexico. The analysis included species diversity, activity peaks and molecular detection of pathogens. Sand flies were collected from November to December 2021 and April to May 2022, during 84 night-traps. The conserved area showed higher numbers and greater species heterogeneity of sand flies as compared with the other sites. The β-diversity analysis revealed that sites disturbed by logging (S1, S2, S3) had greater similarity (90%) in their sand fly species composition than a conserved area (S4) (similarity = 36%). Although none of the specimens were infected with Leishmania, we detected Wolbachia (19.4%) in all four sites, as well as Bartonella (3.25%) only in the disturbed sites. Further studies on the dynamics of sand fly populations and their association with pathogens are necessary.


Structure of the propeptide of prothrombin containing the gamma-carboxylation recognition site determined by two-dimensional NMR spectroscopy.
The propeptides of the vitamin K dependent blood clotting and regulatory proteins contain a gamma-carboxylation recognition site that directs precursor forms of these proteins for posttranslational gamma-carboxylation. Peptides corresponding to the propeptide of prothrombin were synthesized and examined by circular dichroism (CD) and nuclear magnetic resonance spectroscopy (NMR). CD spectra indicate that these peptides have little or no secondary structure in aqueous solutions but that the addition of trifluoroethanol induces or stabilizes a structure containing alpha-helical character. The maximum helical content occurs at 35-40% trifluoroethanol. This trifluoroethanol-stabilized structure was solved by two-dimensional NMR spectroscopy. The NMR results demonstrate that residues -13 to -3 form an amphipathic alpha-helix. NMR spectra indicate that a similar structure is present at 5 degrees C, in the absence of trifluoroethanol. Of the residues previously implicated in defining the gamma-carboxylation recognition site, four residues (-18, -17, -16, and -15) are adjacent to the helical region and one residue (-10) is located within the helix. The potential role of the amphipathic alpha-helix in the gamma-carboxylation recognition site is discussed.


Subinhibitory arsenite concentrations lead to population dispersal in Thiomonas sp.
Biofilms represent the most common microbial lifestyle, allowing the survival of microbial populations exposed to harsh environmental conditions. Here, we show that the biofilm development of a bacterial species belonging to the Thiomonas genus, frequently found in arsenic polluted sites and playing a key role in arsenic natural remediation, is markedly modified when exposed to subinhibitory doses of this toxic element. Indeed, arsenite [As(III)] exposure led to a considerable impact on biofilm maturation by strongly increasing the extracellular matrix synthesis and by promoting significant cell death and lysis within microcolonies. These events were followed by the development of complex 3D-biofilm structures and subsequently by the dispersal of remobilized cells observed inside the previously formed hollow voids. Our results demonstrate that this biofilm community responds to arsenite stress in a multimodal way, enhancing both survival and dispersal. Addressing this complex bacterial response to As(III) stress, which might be used by other microorganisms under various adverse conditions, may be essential to understand how Thiomonas strains persist in extreme environments.


Spouses' attributions for helping: the effects of styles of help-seeking, self-serving bias, and sex.
This paper examines the effects of (a) how clearly the desire for help is presented, (b) self-serving bias, and (c) sex differences on the source of the cause of helping happening (or not) that spouses attribute to a potential helper. In two studies, a total of 138 couples imagined situations in which a person wanted his or her spouse to do a chore. How clearly the desire for help was expressed, whether help was given, and whether it was the participant or the spouse who was expected to help were manipulated. The participants reported their attributions for the cause of helping (or not) to themselves and the spouse on visual analog scales. There was strong support for a proposed interaction between how clearly the desire for help was expressed and whether help was given in predicting spouses' attributions. No support was obtained for a predicted self-serving bias or for a suggestion that men make more self-enhancing attributions than women, the findings were instead congruent with a relationship-enhancing bias. The findings are discussed in terms of the context of housework.


Occult transfixation of the sigmoid colon by suprapubic catheter.
A 75-year old man with severe cerebrovascular disease underwent a routine change of suprapubic catheter three months after first insertion. One day after the catheter was changed, he passed faeculent material in the catheter and became unwell with abdominal pain. The catheter tip was visible per rectum. A CT scan confirmed that the suprapubic catheter had passed into the sigmoid colon. He underwent laparotomy and repair of a colovesical fistula and sigmoid perforation. He made an uncomplicated recovery. Damage to the bowel is a rare but recognized complication of suprapubic catheter insertion. Our patient illustrates that the injury may not become apparent until a change of catheter, and clinicians should bear in mind the possibility of occult bowel damage if patients become unwell after a change of suprapubic catheter.


Activation of phosphatidylinositol 3-kinase, Akt, and mammalian target of rapamycin is necessary for hypoxia-induced pulmonary artery adventitial fibroblast proliferation.
In contrast to cell types in which exposure to hypoxia causes a general reduction of metabolic activity, a remarkable feature of pulmonary artery adventitial fibroblasts is their ability to proliferate in response to hypoxia. Previous studies have suggested that ERK1/2, phosphatidylinositol 3-kinase (PI3K), Akt, and mammalian target of rapamycin (mTOR) are activated by hypoxia and play a role in a variety of cell responses. However, the pathways involved in mediating hypoxia-induced proliferation are largely unknown. Using pharmacological inhibitors, we established that PI3K-Akt, mTOR-p70 ribosomal protein S6 kinase (p70S6K), and EKR1/2 signaling pathways play a critical role in hypoxia-induced adventitial fibroblast proliferation. We found that exposure of serum-starved fibroblasts to 3% O2 resulted in a time-dependent activation of PI3K and transient phosphorylation of Akt. However, activation of PI3K was not required for activation of ERK1/2, implying a parallel involvement of these pathways in the proliferative response of fibroblasts to hypoxia. We found that hypoxia induced significant increases in mTOR, p70S6K, 4E-BP1, and S6 ribosomal protein phosphorylation, as well as dramatic increases in p70S6K activity. The activation of p70S6K/S6 pathway was sensitive to inhibition by rapamycin and LY294002, indicating that mTOR and PI3K/Akt are upstream signaling regulators. However, the magnitude of hypoxia-induced p70S6K activity and phosphorylation suggests involvement of additional signaling pathways. Thus our data demonstrate that hypoxia-induced adventitial fibroblast proliferation requires activation and interaction of PI3K, Akt, mTOR, p70S6K, and ERK1/2 and provide evidence for hypoxic regulation of protein translational pathways in cells exhibiting the capability to proliferate under hypoxic conditions.


Similar Profile and Magnitude of Cognitive Impairments in Focal and Generalized Epilepsy: A Pilot Study.
Introduction: Cognitive impairments in epilepsy are not well-understood. In addition, long-term emotional, interpersonal, and social consequences of the underlying disturbances are important to evaluate. Purpose: To compare cognitive function including language in young adults with focal or generalized epilepsy. In addition, quality of life and self-esteem were investigated. Patients and Methods: Young adults with no primary intellectual disability, 17 with focal epilepsy and 11 with generalized epilepsy participated and were compared to 28 healthy controls. Groups were matched on age (mean = 26 years), sex, and education. Participants were administered a battery of neuropsychological tasks and carried out self-ratings of quality of life, self-esteem, and psychological problems. Results: Similar impairments regarding cognitive function were noted in focal and generalized epilepsy. The cognitive domains tested were episodic long-term memory, executive functions, attention, working memory, visuospatial functions, and language. Both epilepsy groups had lower results compared to controls (effect sizes 0.24-1.07). The total number of convulsive seizures was predictive of episodic long-term memory function. Participants with focal epilepsy reported lower quality of life than participants with generalized epilepsy. Lowered self-esteem values were seen in both epilepsy groups and particularly in those with focal epilepsy. Along with measures of cognitive speed and depression, the total number of seizures explained more than 50% of variation in quality of life. Conclusion: Interestingly, similarities rather than differences characterized the widespread cognitive deficits that were seen in focal and generalized epilepsy, ranging from mild to moderate. These similarities were modified by quality of life and self-esteem. This study confirms the notion that epilepsy is a network disorder.


Comparison of the effectiveness of wireless electronic tracking devices versus traditional paper systems to track victims in a large scale disaster.
We conducted an unblinded experimental comparative trial during a disaster drill involving DMATs using the WIISARD system and traditional paper tracking of casualties. We shadowed the paper work flow to collect data on 40 victims tracked by both systems. WIISARD captured patients as well as the paper system. However, WIISARDwas superior at tracking patient destinations and transporting units. WIISARD proved to be an effective victim tracking system.