The CO2 responsiveness and ventilatory response to leg and arm exercise in female swimmers.
The primary purpose of this study was to compare the ventilatory response to CO2 during rebreathing (delta VI/delta PCO2) among 8 competitive synchronized swimmers (SS), 8 competitive speed swimmers (CS) and 8 recreation swimmers (RS) who acted as controls. A secondary purpose was to study the relationship between delta VI/delta PCO2 and the ventilatory equivalent to the metabolic CO2 output (delta VE/delta VCO2) during graded exercise consisting of treadmill walking and arm cranking. No significant difference was observed amongst the groups in either the intercept or slope of the ventilatory response to increasing alveolar PCO2 was 1.48 +/- 0.33, 2.04 +/- 0.13 and 1.87 +/- 0.18 (mean +/- SEM) in the SS, CS and RS, respectively. The delta VI/delta PCO2 during rebreathing; delta VI/delta PCO2 during rebreathing was not significantly related to the delta VE/delta VCO2 obtained during either treadmill walking or arm cranking. The delta VE/delta VCO2 achieved during treadmill walking was similar for the 3 groups. However, during arm cranking, the delta VE/delta VCO2 was similar for the SS and CS, but both values were significantly lower than the delta VE/delta VCO2 achieved by the RS. It appears that the training regime experienced by the SS and CS did not alter their responsiveness to CO2 during rebreathing. The reduced delta VE/delta VCO2 observed during exercise with the trained limbs (arms) in the SS and CS compared to RS cannot be explained by altered responsiveness to CO2 secondary to training.


In vitro inhibitory activity of citreoviridin against HIV-1 and an HIV-associated opportunist: Candida albicans.
Citreoviridin, a mycotoxin produced by some molds of the genera Penicillium and Aspergillus, inhibits the growth of bacteria of the Bacillus genus. Since significant information was not available on the effects of citreoviridin on eukaryotic cells and viruses, this molecule was tested on CD4+ T-lymphoid cell lines, on HIV-1 and on Candida albicans, which sometimes complicates HIV-infection. Antiviral activity was detected in H9 HTLV IIIB cells, a clone chronically infected by HIV-1. Citreoviridin reduced p24 in the supernatant of H9 HTLV IIIB in a dose-dependent manner with a pharmacological selectivity index of 2.6. In C. albicans, the effects of the mold-derivative were evaluated on some parameters associated with pathogenicity and virulence: cellular proliferation, germ tube production, expression of heat shock mannoproteins, release of proteases and phospholipases. At a 12.5 microM dose, citreoviridin showed a marked inhibitory effect on all parameters analyzed. As regards the mechanism of action, it is possible to hypothesize that the effects of citreoviridin may be due to a reduction of protein synthesis, since it inhibited the replication of HIV-1 at post-integrational stages and reduced the intracellular RNA and protein content in C. albicans.


The Next Frontier: Translational Development of Ubiquitination, SUMOylation, and NEDDylation in Cancer.
Post-translational modifications of proteins ensure optimized cellular processes, including proteostasis, regulated signaling, cell survival, and stress adaptation to maintain a balanced homeostatic state. Abnormal post-translational modifications are associated with cellular dysfunction and the occurrence of life-threatening diseases, such as cancer and neurodegenerative diseases. Therefore, some of the frequently seen protein modifications have been used as disease markers, while others are targeted for developing specific therapies. The ubiquitin and ubiquitin-like post-translational modifiers, namely, small ubiquitin-like modifier (SUMO) and neuronal precursor cell-expressed developmentally down-regulated protein 8 (NEDD8), share several features, such as protein structures, enzymatic cascades mediating the conjugation process, and targeted amino acid residues. Alterations in the regulatory mechanisms lead to aberrations in biological processes during tumorigenesis, including the regulation of tumor metabolism, immunological modulation of the tumor microenvironment, and cancer stem cell stemness, besides many more. Novel insights into ubiquitin and ubiquitin-like pathways involved in cancer biology reveal a potential interplay between ubiquitination, SUMOylation, and NEDDylation. This review outlines the current understandings of the regulatory mechanisms and assay capabilities of ubiquitination, SUMOylation, and NEDDylation. It will further highlight the role of ubiquitination, SUMOylation, and NEDDylation in tumorigenesis.


Oral Disintegration Tablets of Stavudine for HIV Management: A New Technological Approach.
Stavudine oral disintegration tablets were formulated to minimize the bitter taste and to reduce the first-pass hepatic metabolism. The various precompression parameters like the angle of repose, bulk density, compressibility index and Hausner's ratio were determined for the powder blend. In this study, 14 formulations of stavudine oral disintegration tablet were prepared by direct compression method. The tablets were evaluated for weight variation, percentage friability, disintegration time, hardness, wetting time and water absorption ratio. The in vitro dissolution study results of the batch S1 (stavudine+crospovidone+sodium starch glycollate) are encouraging as highest dissolution rate (99.2% in 100 min) and lowest time of disintegration (56 s) was achieved. The in vivo drug release studies were carried out in rabbits and the relative bioavailability of formulation S1 was found to be 2.83 times greater than that of conventional tablets.


Helping children cope with parental mental illness through use of children's literature.
Children of mentally ill parents are often vulnerable, partly because they may not receive adequate nurturing and partly because they feel stigmatized by their parents' disabilities. The author recommends use in treatment of several children's books that show realistic, positive role models of children coping with parental mental illness.


Histologic abnormalities are common in protocol liver allograft biopsies from patients with normal liver function tests.
The utility of protocol liver allograft biopsies remains controversial, particularly in patients with normal liver function tests (LFTs). However, histologic evaluation of these biopsies provides an opportunity to examine the types and severity of liver diseases that can occur in livers with normal clinical and biochemical function. We studied 165 protocol allograft biopsies taken from 100 liver transplant patients at the time of normal LFTs and normal clinical function at 3 to 8 months (n=36), 1 year (n=52), 2 to 3 years (n=54), and 4 to 5 years (n=23). Biopsies were classified as normal, minimal changes (eg, nonaggressive portal or lobular mononuclear inflammation, steatosis <10%), fatty liver disease, recurrent primary liver disease, and transplant-related disease (portal-based rejection or central venulitis, an inflammatory pattern that encompasses perivenular hepatocyte dropout, mononuclear inflammation, pigment-laden macrophages, and variable zone 3 fibrosis). Among these 100 patients, a total of 394 protocol biopsies were performed, and 165 (42%) were taken at the time of normal LFTs and normal clinical function. One hundred twenty-one (73%) were normal or showed minimal/nonspecific changes. Forty-four (27%) showed histologic abnormalities that included fatty liver disease (n=19, nonalcoholic in 18 cases; 13 with mild steatosis, 6 with moderate steatosis, 7 with grade 1/3 steatohepatitic activity, and 2 with stage 1/4 steatohepatitic fibrosis), recurrent primary biliary cirrhosis (n=9; all stage 1/4), recurrent hepatitis C infection (n=6; grade 0/4 in 1, grade 1/4 in 5, stage 0/4 in 4, stage 1/4 in 1, and stage 2/4 in 1), recurrent sarcoidosis (n=1), Ito cell hyperplasia (n=4; marked in 2 and mild in 2), central venulitis (n=10; 5 with mild zone 3 fibrosis or central vein obliteration and 1 with central-portal bridging fibrosis), and mild acute portal rejection (n=2). We judged the histologic changes to be of clinical significance in 19 (11.5%) cases. These results indicate that even at the time of normal clinical and laboratory function, a significant fraction of protocol allograft biopsies harbor histologic (27%) and clinically significant (11.5%) abnormalities. These most commonly include fatty liver disease, low-grade/low-stage recurrent hepatitis C and primary biliary cirrhosis, and central venulitis (including some cases with subsequent fibrosis progression). The data support performance of protocol biopsies to assess allograft status, and provide insight into the types and severity of liver diseases that can smolder in transplanted (and by extension, probably also in native) livers with apparent normal function.


Perfluoroalkyl substances and changes in bone mineral density: A prospective analysis in the POUNDS-LOST study.
Recent studies suggested an inverse association between exposures to perfluoroalkyl substances (PFASs) and bone mineral density (BMD). Whether exposures to PFASs are also associated with changes in BMD has not been examined. Five major PFASs (perfluorooctanesulfonic acid, PFOS; perfluorooctanoic acid, PFOA; perfluorohexanesulfonic acid, PFHxS; perfluorononanoic acid, PFNA; perfluorodecanoic acid, PFDA) and BMD (g/cm2) at six bone sites (spine, total hip, femoral neck, hip intertrochanteric area, hip trochanter, and hip Ward's triangle area) were measured at baseline among 294 participants in the POUNDS-LOST study, a weight-loss trial, of whom a total of 175 participants had BMD measured at both baseline and year 2. Linear regression was used to model the differences or changes in BMD for each SD increment of PFAS concentrations. In a secondary analysis, interactions between PFASs and baseline body mass index (BMI), as well as a BMI-related genetic risk score (GRS) derived from 97 BMI-predicting SNPs were examined in relation to changes in BMD. At baseline, both PFOS and PFOA were significantly associated with lower BMD at several sites. For each SD increase of PFOS, the βs (95% CIs) for BMD were -0.020(-0.037, -0.003) for spine, -0.013(-0.026, 0.001) for total hip, -0.014(-0.028, 0.000) for femoral neck, and -0.013(-0.026, 0.000) for hip trochanter. For PFOA, the corresponding figures were -0.021(-0.038, -0.004) for spine, -0.015(-0.029, -0.001) for total hip, and -0.015(-0.029, -0.002) for femoral neck. After adjusting for baseline covariates and 2-year weight change, higher baseline plasma concentrations of PFOS, PFNA, and PFDA were associated with greater reduction in BMD in the hip; the βs (95% CIs) were -0.005(-0.009, -0.001), -0.006(-0.010, -0.001), and -0.005(-0.009, -0.001), respectively. Similar associations were found in hip intertrochanteric area for all PFASs except PFHxS, with βs ranging from -0.006 for PFOA to -0.008 for PFOS and PFNA. Participants with a higher GRS tended to have less PFAS-related BMD decline in total hip (Pinteraction = 0.005) and the hip intertrochanteric area (Pinteraction = 0.021). There were similar PFAS-related BMD changes by baseline BMI levels, although the interactions did not achieve statistical significance. This study demonstrated that higher plasma PFAS concentrations were not only associated with a lower BMD at baseline, but also a faster BMD loss in a weight-loss trial setting. Genetic predisposition to larger body size may somewhat attenuate the deleterious effects of PFASs on BMD. Further exploration of the possible impact of PFAS exposures on bone density is warranted.


Health Care for People with Disabilities in the Unified Health System in Brazil: A Scoping Review.
People with disabilities have greater need for healthcare on average, but often face barriers when accessing these services. The Brazilian government launched the National Health Policy for People with Disabilities (PNSPD) in 2002 to address this inequality. PNSPD has six areas of focus: quality of life, impairment prevention, comprehensive health care, organization and functioning of health services, information mechanisms, and training of human resources. The aim of this article was to undertake a scoping review to assess the evidence on the experience of people with disabilities in Brazil with respect to the six themes of the PNSPD. The scoping review included articles published between 2002 and 2019, from four electronic databases: PUBMED/MEDLINE, LILACS, Science Direct, and Scielo. In total, 8076 articles were identified, and after review of titles, abstracts, and full texts by two independent reviewers, 98 were deemed eligible for inclusion. The evidence was relatively limited in availability and scope. However, it consistently showed large gaps in delivery of healthcare to people with disabilities across the six dimensions considered. There was lack of actions aimed at promoting quality of life; insufficient professional training about disability; little evidence on the health profile of people with disabilities; large gaps in the availability of care due to widespread physical, informational, and attitudinal barriers; and poor distribution of the supply and integration of services. In conclusion, the policy framework in Brazil is supportive of the inclusion of people with disabilities in health services; however, large inequalities remain due to poor implementation of the policy into practice.


Safety and benefit of ambulation within 24 hours in elderly patients undergoing lumbar fusion: propensity score matching study of 882 patients.
Elderly patients are less likely to recover from lumbar spine fusion (LSF) as rapidly compared with younger patients. However, there is still a lack of research on the effect of early ambulation on elderly patients undergoing LSF surgery for lumbar degenerative disorders. To evaluate the safety and benefit of ambulation within 24 hours in elderly patients who underwent LSF. A retrospective study. Consecutive patients (aged 65 and older) who underwent elective transforaminal lumbar interbody fusion surgery for degenerative disorders from January 2019 to October 2022. Outcome measures included postoperative complications, postoperative drainage (ml), laboratory test data, length of hospital stay [LOS], readmission and reoperation within 3 months. Early ambulation patients (ambulation within 24 h after surgery) were propensity-score matched 1:1 to a delayed ambulation patients (ambulation at a minimum of 48 h postoperatively) based on age, intraoperative blood loss, and number of fused segments. The incidence of postoperative adverse events (AEs, including rates of complications, readmission, and prolonged LOS) and the average LOS were used to assess the safety and benefit of early ambulation, respectively. Multivariable regression analysis was performed to assess the association between early ambulation and postoperative AEs. The risk factors for delayed ambulation were also determined using multivariable logistic analyses. A total of 998 patients with LSF surgery were reviewed in this study. After excluding 116 patients for various reasons, 882 patients (<24 hours: N = 350, 24-48 hours: N = 230, and >48 hours: N= 302) were included in the final analysis. After matching, gender, BMI, preoperative comorbidities, laboratory test data and surgery-related variables were comparable between the groups. The incidence of postoperative AEs was significantly lower in the EA group (44444.3% vs. 64.0%, p < .001). The average postoperative LOS of the EA group was 2 days shorter than the DA group (6 .5d vs. 8.5d, p <.001). Patients in the EA group had a significantly lower rate of prolonged LOS compared to the DA group (33535.1% vs. 55.3%, p <.001). There was no significant difference in postoperative drainage volumes between the two groups. Multivariable analysis identified older age (OR 1.07, p <.001), increased intraoperative EBL (OR 1.002, p = 0.001), and higher international normalization ratio (OR 10.57, p = 0.032) as significant independent risk factors for delayed ambulation. Ambulation within 24 hours after lumbar spine fusion surgery is independently associated fewer adverse events and shorter hospital stays in elderly patients. Implementing the goal of ambulation within 24 hours after LSF surgery into enhanced recovery after surgery protocols for elderly patients seems appropriate. Older age, increased intraoperative blood loss and worse coagulation function are associated with delayed ambulation.


Septal flash and septal rebound stretch have different underlying mechanisms.
Abnormal left-right motion of the interventricular septum in early systole, known as septal flash (SF), is frequently observed in patients with left bundle branch block (LBBB). Transseptal pressure gradient and early active septal contraction have been proposed as explanations for SF. Similarities in timing (early systole) and location (septum) suggest that SF may be related to septal systolic rebound stretch (SRSsept). We aimed to clarify the mechanisms generating SF and SRSsept. The CircAdapt computer model was used to isolate the effects of timing of activation of the left ventricular free wall (LVFW), right ventricular free wall (RVFW), and septum on SF and SRSsept. LVFW and septal activation times were varied by ±80 ms relative to RVFW activation time. M-mode-derived wall motions and septal strains were computed and used to quantify SF and SRSsept, respectively. SF depended on early activation of the RVFW relative to the LVFW. SF and SRSsept occurred in LBBB-like simulations and against a rising transseptal pressure gradient. When the septum was activated before both LVFW and RVFW, no SF occurred despite the presence of SRSsept. Computer simulations therefore indicate that SF and SRSsept have different underlying mechanisms, even though both can occur in LBBB. The mechanism of leftward motion during SF is early RVFW contraction pulling on and straightening the septum when unopposed by the LVFW. SRSsept is caused by late LVFW contraction following early contraction of the septum. Changes in transseptal pressure gradient are not the main cause of SF in LBBB.


Fabrication and properties of free-standing c60 membranes.
Van der Waals forces that bind C(60) molecular solids are found to be sufficiently strong to allow the reproducible fabrication of free-standing C(60) membranes on (100) silicon wafers. Membranes, 2000 to 6000 angstroms thick, were fabricated by a modified silicon micro-machining process and were found to be smooth, flat, and mechanically robust. An important aspect of the silicon-compatible fabrication procedure is the demonstration that C(60) films can be uniformly and nondestructively thinned in a CF(4) plasma. Young's modulus and fracture strength measurements were made on membranes with areas larger than 6 millimeters by 6 millimeters. It may be possible to use C(60), membranes for physical property measurements and applications.


Evaluating if an Advance Care Planning Intervention Promotes Do-Not-Resuscitate Orders by Facilitating Accurate Prognostic Awareness.
Issuing do-not-resuscitate (DNR) orders has seldom been an outcome in randomized clinical trials of advance care planning (ACP) interventions. The aim of this study was to examine whether an ACP intervention facilitating accurate prognostic awareness (PA) for patients with advanced cancer was associated with earlier use of DNR orders. Participants (n=460) were randomly assigned 1:1 to the experimental and control arms, with 392 deceased participants constituting the final sample of this secondary analysis study. Participants in the intervention and control arms had each received an intervention tailored to their readiness for ACP/prognostic information and symptom-management education, respectively. Effectiveness in promoting a DNR order by facilitating accurate PA was determined by intention-to-treat analysis using multivariate logistic regression with hierarchical linear modeling. At enrollment in the ACP intervention and before death, 9 (4.6%) and 8 (4.1%) participants and 168 (85.7%) and 164 (83.7%) participants in the experimental and control arms, respectively, had issued a DNR order, without significant between-arm differences. However, participants in the experimental arm with accurate PA were significantly more likely than participants in the control arm without accurate PA to have issued a DNR order before death (adjusted odds ratio, 2.264; 95% CI, 1.036-4.951; P=.041). Specifically, participants in the experimental arm who first reported accurate PA 31 to 90 days before death were significantly more likely than their counterparts in the control arm who reported accurate PA to have issued a DNR order in the next wave of assessment (adjusted odds ratio, 13.365; 95% CI, 1.989-89.786; P=.008). Both arms issued DNR orders close to death (median, 5-6 days before death). Our ACP intervention did not promote the overall presence of a DNR order. However, our intervention facilitated the issuance of NDR orders before death among patients with accurate PA, especially those who reported accurate PA 31 to 90 days before death, but it did not facilitate the issuance of DNR orders earlier than their counterparts in the control arm.ClinicalTrial.gov Identification: NCT01912846.


[Leadership in critical care settings: reflections and challenges for the Brazilian Nursing].
The objective of this study was to reflect on the leadership, especially in the intensive care nursing, based on the prospect of nurses, teachers, nursing students and in the literature on the subject. The leadership of the nurse in intensive care units faces challenges and paradoxes and is present in the planning of care, in the moments of ethical and bioethical conflicts, in the supervision of care, in capacity of making decisions quickly and safety, in knowing deal with the issues involving the process of living and dying.


[Evaluation of functional brain condition by EEG biopotential power asymmetry].
The fluctuations of functional brain condition level are evaluated by interhemispheric asymmetry of bioelectric activity power that is a consequence of both hemispheres interplay and involvement of nonspecific brain system in this process. Using neurological examination and electroencephalography, functional CNS condition analysis in patients with chronic brain ischemia and healthy controls was carried out. The results were compared to the indices of interhemispheric asymmetry of biopotential power (IHABP). Different functional CNS condition of both the patients and controls was characterized by different ratio of IHABP for fore- and back brain cortical regions. Controls exhibited a predominance of IHABP for frontal neocortex areas comparing to back ones. In the patients, the identical IHABP indices were detected for both neocortex areas.


Graft-induced dyskinesias in Parkinson's disease: High striatal serotonin/dopamine transporter ratio.
Graft-induced dyskinesias are a serious complication after neural transplantation in Parkinson's disease. One patient with Parkinson's disease, treated with fetal grafts 14 years ago and deep brain stimulation 6 years ago, showed marked improvement of motor symptoms but continued to suffer from OFF-medication graft-induced dyskinesias. The patient received a series of clinical and imaging assessments. Positron emission tomography and single-photon emission computed tomography 14 years posttransplantation revealed an elevated serotonin/dopamine transporter ratio in the grafted striatum compatible with serotonergic hyperinnervation. Inhibition of serotonin neuron activity by systemic administration of a 5-HT(1A) agonist suppressed graft-induced dyskinesias. Our data provide further evidence that serotonergic neurons mediate graft-induced dyskinesias in Parkinson's disease. Achieving a normal striatal serotonin/dopamine transporter ratio following transplantation of fetal tissue or stem cells should be necessary to avoid the development of graft-induced dyskinesias.


End-of-life Decision-making for People in a Minimally Conscious State: A Review of the Application of the Mental Capacity Act 2005.
The last 15 years has seen clarification of the terminology used to describe prolonged disorders of consciousness within the United Kingdom leading to the emergence of a new diagnosis - minimally conscious state (MCS) in 2002. MCS is distinct from vegetative states, in that a person demonstrates wakefulness with some degree of minimal awareness. The Mental Capacity Act (MCA) 2005 in England and Wales provides a legal framework for assessing an individuals' capacity to make decisions for themselves. The Act also authorizes others to make decisions on behalf of an individual who is assessed as lacking capacity in their best interests. The Act has an accompanying Code of Practice which provides guidance and a best interests "test" to be applied when assessing best interests. Since the advent of the Act, approximately two cases each year go to the Court of Protection for final decisions regarding end-of-life care in people in an MCS. Currently, any decision involving the withdrawal of clinically assisted nutrition and hydration (CANH) for people in an MCS must be referred to the court. In each case, the courts analyze the application of the Act which has become central in the court's decision-making process, particularly when assessing best interests. This article provides an overview of key MCA sections applied in such end-of-life MCS cases and reviews seminal cases elucidating how the Act has been applied. It further describes the evolution of how courts have interpreted the doctrine of best interests when considering withholding or withdrawing CANH and other life-sustaining treatments.


Leuprorelin depot injection: patient considerations in the management of prostatic cancer.
Hormone therapy is well established for treating patients with prostate cancer and remains the mainstay of the treatment of metastatic and locally advanced disease, this article reviews the rationale for its use, its different forms, and complications and controversies still surrounding some of its modalities. Availability of long-acting synthetic luteinizing hormone-releasing hormone (LHRH) agonists revolutionized the hormonal treatment of prostate cancer, and helped to avoid the emotional and psychological effects related to surgical castration. The depot formula has gained wide acceptance from both patients and physicians. This review emphasizes the newer, long-acting formula, leuprorelin (leuprolide acetate), especially the 6-month formula, its advantage over over shorter-acting depot products, and its potential to become a standard of care for patients eligible for androgen deprivation therapy.


Suppression of matrix metalloproteinases inhibits establishment of ectopic lesions by human endometrium in nude mice.
Matrix metalloproteinases of the stromelysin family are expressed in the human endometrium as a consequence of cellular events during the menstrual cycle that require extracellular matrix remodeling. We have recently documented the presence of these enzymes in lesions of endometriosis, a benign disease that presents as persistent ectopic sites of endometrial tissue, usually within the peritoneal cavity. Endometriosis can develop after retrograde menstruation of endometrial tissue fragments, and establishment of ectopic sites within the peritoneal cavity requires breakdown of extracellular matrix. To examine whether matrix metalloproteinases might contribute to the steroid-dependent epidemiology and cellular pathophysiology of endometriosis, we have developed an experimental model of endometriosis using athymic nude mice as recipients of human endometrial tissue. Our results demonstrate that estrogen treatment of human endometrial tissue in organ culture maintains secretion of matrix metalloproteinases, and promotes establishment of ectopic peritoneal lesions when injected into recipient animals. In contrast, suppressing metalloproteinase secretion in vitro with progesterone treatment, or blocking enzyme activity with a natural inhibitor of metalloproteinases, inhibits the formation of ectopic lesions in this experimental model.


Photoreceptor processing improves salience facilitating small target detection in cluttered scenes.
Target detection amidst clutter is a challenging task for both natural and artificial vision, yet one solved at the level of neurons in the 3rd optic ganglion of insects. These neurons are capable of responding to the motion of small objects, even against complex moving backgrounds. While the basic physiology has been investigated, little is known about how these cells are able to reject background motion while robustly responding to such small stimuli. By recording intracellularly from fly photoreceptors stimulated with natural image sequences containing a target viewed against a complex moving background, we show that the process of target detection begins at the earliest stages of vision. The temporal processing by photoreceptors alone, in the absence of any spatial interactions, improved the discrimination of targets (essentially a spatial task) by around 70%. This enhancement of target salience can be explained by elaborate models of photoreceptor temporal non-linear dynamics. The application of the functional principals outlined in this work could be utilized in areas such as robotics and surveillance, medical imaging, or astronomy, anywhere it is necessary to detect a small item from a cluttered surround.


Increase in histidine decarboxylase activity in tissues of mice bearing Colon-26 tumor cells.
The changes in histidine decarboxylase (HDC) activity, histamine and tele-methylhistamine contents were examined in tissues of mice after the inoculation of Colon-26 tumor cells subcutaneously into the lower back. The HDC activity in the spleen of mice increased significantly 14 days after the inoculation of Colon-26 and the increase in HDC activity continued for up to 28 days. However, the histamine content in the spleen of tumor-bearing mice was not changed significantly during the observation period. In the following experiments, two subclones of the Colon-26 cell line, cachexia-inducing clone-20 and non cachexia-inducing clone-5, were used and the induction of HDC activity in mice was examined in four tissues, spleen, lung, liver and kidney. Both clone-20 and clone-5 induced the increase in HDC activity to the same extent in the spleen and lung, but not in the liver and kidney. As observed using the Colon-26 original cell line, the histamine contents in the four tissues of tumor-bearing mice were not different from those in the control mice. In contrast, the levels of tele-methylhistamine, one of the major catabolites of histamine, in the tumor-bearing mice increased significantly compared with the control mice in all four tissues examined. There was a correlation between the increase in tele-methylhistamine level and the increase in HDC activity in the tissues. A histological study indicated that the tissue mast cells were not increased in spleen and lung of tumor-bearing mice. These findings indicated that the increase in HDC activity in the spleen and lung occurred in parallel with the growth of inoculated tumor cells in mice and suggested that the cells other than mast cells may be involved in the increase in HDC activity. The tumor-bearing state produced histamine with a high turnover rate in the mouse tissues, especially in the spleen and lung.


Quantitation of brown adipose tissue perfusion in transgenic mice using near-infrared fluorescence imaging.
Brown adipose tissue (BAT; brown fat) is the principal site of adaptive thermogenesis in the human newborn and other small mammals. Of paramount importance for thermogenesis is vascular perfusion, which controls the flow of cool blood in, and warmed blood out, of BAT. We have developed an optical method for the quantitative imaging of BAT perfusion in the living, intact animal using the heptamethine indocyanine IR-786 and near-infrared (NIR) fluorescent light. We present a detailed analysis of the physical, chemical, and cellular properties of IR-786, its biodistribution and pharmacokinetics, and its uptake into BAT. Using transgenic animals with homozygous deletion of Type II iodiothyronine deiodinase, or homozygous deletion of uncoupling proteins (UCPs) 1 and 2, we demonstrate that BAT perfusion can be measured noninvasively, accurately, and reproducibly. Using these techniques, we show that UCP -1/-2 knockout animals, when compared to wild-type animals, have a higher baseline perfusion of BAT but a similar maximal response to beta 3-receptor agonist. These results suggest that compensation for UCP deletion is mediated, in part, by the control of BAT perfusion. Taken together, BAT perfusion can now be measured noninvasively using NIR fluorescent light, and pharmacological modulators of thermogenesis can be screened at relatively high throughput in living animals.


Structure of Arabidopsis thaliana Rubisco activase.
The CO2-fixing enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is inactivated by the formation of dead-end complexes with inhibitory sugar phosphates. In plants and green algae, the ATP-dependent motor protein Rubisco activase restores catalytic competence by facilitating conformational changes in Rubisco that promote the release of the inhibitory compounds from the active site. Here, the crystal structure of Rubisco activase from Arabidopsis thaliana is presented at 2.9 Å resolution. The structure reveals an AAA+ two-domain structure. More than 100 residues in the protein were not visible in the electron-density map owing to conformational disorder, but were verified to be present in the crystal by mass spectrometry. Two sulfate ions were found in the structure. One was bound in the loop formed by the Walker A motif at the interface of the domains. A second sulfate ion was bound at the N-terminal end of the first helix of the C-terminal domain. The protein packs in a helical fashion in the crystal, as observed previously for Rubisco activase, but differences in the helical pitch indicate flexibility in the packing of the protein.


Recent Developments in Semiconducting Polymer Dots for Analytical Detection and NIR-II Fluorescence Imaging.
In recent years, semiconducting polymer dots (Pdots) have attracted enormous attention in applications from fundamental analytical detection to advanced deep-tissue bioimaging due to their ultrahigh fluorescence brightness with excellent photostability and minimal cytotoxicity. Pdots have therefore been widely adopted for a variety types of molecular sensing for analytical detection. More importantly, the recent development of Pdots for use in the optical window between 1000 and 1700 nm, popularly known as the "second near-infrared window" (NIR-II), has emerged as a class of optical transparent imaging technology in the living body. The advantages of the NIR-II region over the traditional NIR-I (700-900 nm) window in fluorescence imaging originate from the reduced autofluorescence, minimal absorption and scattering of light, and improved penetration depths to yield high spatiotemporal images for biological tissues. Herein, we discuss and summarize the recent developments of Pdots employed for analytical detection and NIR-II fluorescence imaging. Starting with their preparation, the recent developments for targeting various analytes are then highlighted. After that, the importance of and latest progress in NIR-II fluorescence imaging using Pdots are reported. Finally, perspectives and challenges associated with the emergence of Pdots in different fields are given.


Moesin, like ezrin, colocalizes with actin in the cortical cytoskeleton in cultured cells, but its expression is more variable.
The band 4.1 superfamily of proteins show approx. 30% sequence identity in their amino-terminal region to the membrane binding domain of erythrocyte band 4.1. Within this superfamily are three members, ezrin, radixin and moesin, that show approx. 75% overall sequence identity. A comparison of the domain structure and intracellular localization of ezrin and moesin in cultured cells is reported here. Limited proteolytic digestion of ezrin or moesin yields a relatively stable 32 kDa domain derived from the amino-terminal region that is homologous to the protease-resistant membrane binding domain of erythrocyte band 4.1. The remaining regions of the two proteins give rise to very different fragments, suggesting that the secondary/tertiary structures of the two proteins are different in these regions. We have generated polyclonal antibodies that discriminate between ezrin and moesin, and do not react with radixin. All cultured cell lines investigated contain ezrin, whereas moesin is variably expressed. Cells that contain both ezrin and moesin show a very similar pattern: both proteins are enriched and colocalize with actin in cell surface structures. Ezrin is also detected in the cytoplasm. In cells with few or no surface structures, both proteins show a patchy distribution in regions of the cell that contain fine networks of actin filaments. No staining of focal contacts or adherens junctions was observed. These results, together with those of others, lead to the conclusion that, of the members of this protein family, only radixin is an authentic component of adherens junctions and focal contacts. Ezrin and moesin are both found in cell surface structures after treatment of human A431 cells with epidermal growth factor, and ezrin, but not moesin, becomes phosphorylated on tyrosine. This study shows that ezrin and moesin have a similar subcellular distribution in cultured cells, yet are distinguishable in their expression, structure and ability to serve as a kinase substrate.


Radiographic evaluation of pulmonary abnormalities after Aelurostrongylus abstrusus inoculation in cats.
Bronchial and focal alveolar lung disease was present in cats 3 to 6 weeks after Aelurostrongylus inoculation. More generalized alveolar lung disease was seen during first-stage larval production 5 to 21 weeks after inoculation. Bronchial and interstitial disease entities were seen on radiographs after partial resolution of alveolar disease. These changes were observed 17 to 40 weeks after inoculation.


Characterization of the ecological interactions of Roundup Ready 2 Yield® soybean, MON 89788, for use in ecological risk assessment.
As part of an ecological risk assessment, Roundup Ready 2 Yield® soybean (MON 89788) was compared to a conventional control soybean variety, A3244, for disease and arthropod damage, plant response to abiotic stress and cold, effects on succeeding plant growth (allelopathic effects), plant response to a bacterial symbiont, and effects on the ability of seed to survive and volunteer in a subsequent growing season. Statistically significant differences between MON 89788 and A3244 were considered in the context of the genetic variation known to occur in soybean and were assessed for their potential impact on plant pest (weed) potential and adverse environmental impact. The results of these studies revealed no effects of the genetic modification that would result in increased pest potential or adverse environmental impact of MON 89788 compared with A3244. This paper illustrates how such characterization studies conducted in a range of environments where the crop is grown are used in an ecological risk assessment of the genetically modified (GM) crop. Furthermore, risk assessors and decision makers use this information when deciding whether to approve a GM crop for cultivation in-or grain import into-their country.


Large symptomatic carotid body tumor resection aided by preoperative embolization and mandibular subluxation.
Carotid body tumors (CBT) are rare and usually benign neoplasms (60%-90%), originating from the mesoderm and neural ectoderm. In view of the extensive and unrelenting growth of unresected CBT, encasing vital neurovascular structures, and the significant incidence of malignancy (> or = 10%), surgical excision is the standard treatment of choice. Despite progress in CBT imaging and surgical technique, cranial nerve deficit, stroke, and death continue to affect 10% to 40% of patients undergoing curative surgical resection, particularly in large tumors proximal to the skull base. In such cases, CBT shrinkage by preoperative embolization, improved surgical access utilizing mandibular subluxation, and electroencephalographic monitoring combined with meticulous surgical technique may enable curative tumor resection, without prohibitive morbidity. In light of associated disability, preoperative acknowledgment of the ever-present substantial risk of cranial nerve injury cannot be overemphasized. We report on a patient with a large symptomatic CBT treated surgically with the aid of mandibular subluxation and preoperative embolization.


Protective Cerium Oxide Coating Promoted Ce-Doping and Reconstruction of High-Valence NiFe Sulfide toward Robust Overall Water Splitting.
Active and stable electrocatalysts toward oxygen evolution reaction (OER) are essential for alkaline water splitting. Herein, an efficient and durable high-valence NiFe-based OER electrocatalyst is developed, featuring a protective CeO2- x coating to prevent the corrosion of carbon substrates during oxidative OER operation, ensuring excellent catalyst stability. The incorporation of a CeO2- x coating also leads to the formation of a Ce-doped NiFe sulfide catalyst. The Ce modulator enables the dynamic transformation of NiFe sulfide into highly active (oxy)hydroxide species with high-valence Ni sites and enhanced Ni─O covalency, thereby improving its OER catalytic activity. Accordingly, the prepared NiFeS2 /CeO2- x /CC catalyst achieves enhanced OER activity with an overpotential of 260 mV at 100 mA cm-2 in 1.0 m KOH. Moreover, the catalyst achieves 100 mA cm-2 current density at an overpotential of 187 mV for the hydrogen evolution reaction. The anion exchange membrane water electrolyzer reached 500 mA cm-2 at 1.73 V cell voltage with excellent stability for 500 h continuous operation. This study demonstrates a promising approach for the fabrication of robust water-splitting electrocatalysts.


Anti-influenza active and low toxic N-phenyl-substituted β-amidoamidines structurally related to natural antibiotic amidinomycin.
A set of racemic N-phenyl-substituted β-amidoamidines hydrochlorides 4, which are structurally related to natural antiviral agent amidinomycin (1), was synthesized in four steps starting from methacryloyl anilide (5). In the final step of the synthetic route, an uncommon monoacylation of β-aminoamidine 8 at the less reactive β-phenylamino-group took place. To rationalize this result, a mechanism which involves initial acylation at the more active amidine-function followed by intramolecular acyl-group transfer to β-phenylamino-group was suggested. All three β-amidoamidines 4d-f bearing long linear aliphatic chain (from n-C8H17 to n-C12H25) revealed significant in vitro activity against influenza A virus (H3N2) and modest cytotoxicity. The in vitro antiviral potency of 4d,e is 6-20 times greater than that of commercial rimantadine with lower EC50 values and higher therapeutic index. The non-toxic in vivo compounds 4d-f showed a beneficial protective effect in influenza A (H3N2) infected mice.


Cross-species genetics converge to TLL2 for mouse avoidance behavior and human bipolar disorder.
Interspecies genetic analysis of neurobehavioral traits is critical for identifying neurobiological mechanisms underlying psychiatric disorders, and for developing models for translational research. Recently, after screening a chromosome substitution strain panel in an automated home cage environment, chromosomes 15 and 19 were identified in female mice for carrying genetic loci that contribute to increased avoidance behavior (sheltering preference). Furthermore, we showed that the quantitative trait locus (QTL) for baseline avoidance behavior on chromosome 15 is homologous with a human linkage region for bipolar disorder (8q24). Similarly, we now performed comparative analysis on the QTL for avoidance behavior found on chromosome 19 and correspondingly revealed an overlap of the mouse interval and human homologous region 10q23-24, which has been previously linked to bipolar disorders. By means of a comparative genetic strategy within the human homologous region, we describe an association for TLL2 with bipolar disorder using the genome-wide association study (GWAS) data set generated by the Wellcome Trust Case Control Consortium (WTCCC). On the basis of genetic homology and mood stabilizer sensitivity, our data indicate the intriguing possibility that mouse home cage avoidance behavior may translate to a common biochemical mechanisms underlying bipolar disorder susceptibility. These findings pave new roads for the identification of the molecular mechanisms and novel treatment possibilities for this psychiatric disorder, as well as for the validity of translational research of associated psychiatric endophenotypes.


Tissue culture-specific expression of a naturally occurring tobacco feedback-insensitive anthranilate synthase.
A cDNA and corresponding promoter region for a naturally occurring, feedback-insensitive anthranilate synthase (AS) alpha-subunit gene, ASA2, has been isolated from an unselected, but 5-methyl-tryptophan-resistant (5MTr), tobacco (Nicotiana tabacum) cell line (AB15-12-1). The ASA2 cDNA contains a putative transit peptide sequence, and Southern hybridization shows that more than one closely related sequence is present in the tobacco genome. The ASA2 cDNA complemented a trpE nonsense mutant Escherichia coli strain, allowing growth on 300 microm 5MT-containing minimal medium without tryptophan, and cell extracts contained feedback-insensitive AS activity. The 5MTr was lost when the E. coli strain was transformed with an ASA2 site-directed mutant (phenylalanine-107-arginine-108 --> serine-107-glutamine-108). Identical nucleotide sequences encoding the phenylalanine-107-arginine-108 region have been found in polymerase chain reaction-amplified 326-bp ASA2 genomic fragments of wild-type (5-methyl-tryptophan-sensitive [5MTs]) tobacco and a progenitor species. High-level ASA2 transcriptional expression was detected only in 5MTr-cultured cells, not in 5MTs cells or in plants. Promoter studies indicate that tissue specificity of ASA2 is controlled by the promoter region between -2252 and -607. Since the ASA2 promoter sequences are not substantially different in the 5MTr and 5MTs lines, the increased levels of ASA2 mRNA in the 5MTr lines are most likely due to changes in a regulatory gene affecting ASA2 expression.


Backbones of folded proteins reveal novel invariant amino acid neighborhoods.
Folding of naturally occurring proteins has eluded a universal molecular level explanation till date. Rather, there is an abundance of diverse views on dominant factors governing protein folding. Through rigorous analyses of several thousand crystal structures, we observe that backbones of folded proteins display some remarkable invariant features. Folded proteins are characterized by spatially well-defined, distance dependent, and universal, neighborhoods of amino acids which defy any of the conventionally prevalent views. These findings present a compelling case for a newer view of protein folding which takes into account solvent mediated and amino acid shape and size assisted optimization of the tertiary structure of the polypeptide chain to make a functional protein.


Self-control or social control - what determines sleep hygiene in bed-sharing couples?
To investigate intimate partners' impact on sleep hygiene with focus on the temporal dimension and differential predictors of sleep hygiene in co-sleepers and individual sleepers. Habitual co-sleepers and individual sleepers (n=102) completed a cross-sectional, self-report, in-lab, digital survey on sleep hygiene, habitual sleeping arrangement, self-control, depressiveness, and sociodemographic parameters. The relationship between sleeping arrangement and sleep hygiene in co-sleepers was time-dependent with an initial steep incline and a subsequent plateau at approximately one year of co-sleeping routine. Co-sleepers with more than one year of unaltered sleeping arrangement had significantly better sleep hygiene than co-sleepers with less than one-year or individual sleepers. More than one-year continuity of the sleeping arrangement moreover robustly predicted sleep hygiene in co-sleepers whereas self-control was the dominant predictor in individual sleepers. Amongst others, our findings support the idea that insomnia treatment could be improved by becoming sensitive to the habitual sleeping arrangement.


Funding the Educational Mission in Neurology.
Although it is self-evident that education in neurology is important and necessary, how to fund the educational mission is a frequent challenge for neurology departments and clinicians. Department chairs often resort to a piecemeal approach, cobbling together funding for educators from various sources, but frequently falling short. Here, we review the various sources available to fund the educational mission in neurology, understanding that not every department will have access to every source. We describe the multiple different teaching models and formats used by the modern student and educator and their associated costs, some of which are exorbitant. We discuss possible nonfinancial incentives, including pathways to promotion, educational research, and other awards and recognition. Neurological education is commonly underfunded, and departments and institutions must be nimble and creative in finding ways to fund the time and effort of educators.


Non-diffractive Bessel-Gauss beams for the detection of rotating object free of obstructions.
Bessel-Gauss beams carrying orbital angular momentum are widely known for their non-diffractive or self-reconstructing performance, and have been applied in lots of domains. Here we demonstrate that, by illuminating a rotating object with high-order Bessel-Gauss beams, a frequency shift proportional to the rotating speed and the topological charge is observed. Moreover, the frequency shift is still present once an obstacle exists in the path, in spite of the decreasing of received signals. Our work indicates the feasibility of detecting rotating objects free of obstructions, and has potential as obstruction-immune rotation sensors in engine monitoring, aerological sounding, and so on.


Transzonular Triamcinolone-Moxifloxacin Versus Topical Drops for the Prophylaxis of Postoperative Inflammation After Cataract Surgery.
Purpose: To compare the safety and efficacy of an intraoperative transzonular injection of triamcinolone-moxifloxacin (Imprimis' Tri-Moxi 15 mg/1 mg/mL) to topical drops in patients undergoing cataract surgery. Methods: Between January 2017 and October 2017, patients undergoing phacoemulsification cataract surgery by a single surgeon at the Penn State Eye Center were offered a single intraoperative injection of transzonular triamcinolone-moxifloxacin in lieu of perioperative drops. Between November 2017 and July 2018, the transzonular injection was not offered, and all patients who underwent surgery by the same surgeon received a drop regimen consisting of polymyxin b/trimethoprim and prednisolone acetate 1%. Patients were seen 1 day, 1 week, and 6-9 weeks postoperatively. Outcome measures included incidence of intraoperative complications, postoperative breakthrough inflammation, cystoid macular edema, and infectious sequela. Results: Of the 198 eyes, 99 from 73 patients received the injection and 99 from 82 patients received topical drops. One (1%) intraoperative posterior capsule tear occurred in each group. Eleven (11.1%) eyes in the injection group and 3 (3%) in the drop group experienced symptomatic breakthrough inflammation necessitating treatment (P = 0.0488). One (1%) eye in the injection group and zero (0%) in the drop group developed clinically significant macular edema (P = 1.0). No instances of elevated intraocular pressure or infectious sequela occurred in either group (P = 1.0). Conclusions: Transzonular injection of triamcinolone-moxifloxacin may be associated with an increased incidence of breakthrough inflammation compared to topical drops.


Dysregulation of Subcutaneous White Adipose Tissue Inflammatory Environment Modelling in Non-Insulin Resistant Obesity and Responses to Omega-3 Fatty Acids - A Double Blind, Randomised Clinical Trial.
Obesity is associated with enhanced lipid accumulation and the expansion of adipose tissue accompanied by hypoxia and inflammatory signalling. Investigation in human subcutaneous white adipose tissue (scWAT) in people living with obesity in which metabolic complications such as insulin resistance are yet to manifest is limited, and the mechanisms by which these processes are dysregulated are not well elucidated. Long chain omega-3 polyunsaturated fatty acids (LC n-3 PUFAs) have been shown to modulate the expression of genes associated with lipid accumulation and collagen deposition and reduce the number of inflammatory macrophages in adipose tissue from individuals with insulin resistance. Therefore, these lipids may have positive actions on obesity associated scWAT hypertrophy and inflammation. To evaluate obesity-associated tissue remodelling and responses to LC n-3 PUFAs, abdominal scWAT biopsies were collected from normal weight individuals and those living with obesity prior to and following 12-week intervention with marine LC n-3 PUFAs (1.1 g EPA + 0.8 g DHA daily). RNA sequencing, qRT-PCR, and histochemical staining were used to assess remodelling- and inflammatory-associated gene expression, tissue morphology and macrophage infiltration. Obesity was associated with scWAT hypertrophy (P < 0.001), hypoxia, remodelling, and inflammatory macrophage infiltration (P = 0.023). Furthermore, we highlight the novel dysregulation of Wnt signalling in scWAT in non-insulin resistant obesity. LC n-3 PUFAs beneficially modulated the scWAT environment through downregulating the expression of genes associated with inflammatory and remodelling pathways (P <0.001), but there were altered outcomes in individuals living with obesity in comparison to normal weight individuals. Our data identify dysregulation of Wnt signalling, hypoxia, and hypertrophy, and enhanced macrophage infiltration in scWAT in non-insulin resistant obesity. LC n-3 PUFAs modulate some of these processes, especially in normal weight individuals which may be preventative and limit the development of restrictive and inflammatory scWAT in the development of obesity. We conclude that a higher dose or longer duration of LC n-3 PUFA intervention may be needed to reduce obesity-associated scWAT inflammation and promote tissue homeostasis. www.isrctn.com, identifier ISRCTN96712688.


Simultaneous Dual Encoding of Three-Dimensional Structures by Light-Induced Modular Ligation.
A highly efficient strategy for the simultaneous dual surface encoding of 2D and 3D microscaffolds is reported. The combination of an oligo(ethylene glycol)-based network with two novel and readily synthesized monomers with photoreactive side chains yields two new photoresists, which can be used for the fabrication of microstructures (by two-photon polymerization) that exhibit a dual-photoreactive surface. By combining both functional photoresists into one scaffold, a dual functionalization pattern can be obtained by a single irradiation step in the presence of adequate reaction partners based on a self-sorting mechanism. The versatility of the approach is shown by the dual patterning of halogenated and fluorescent markers as well as proteins. Furthermore, we introduce a new ToF-SIMS mode ("delayed extraction") for the characterization of the obtained microstructures that combines high mass resolution with improved lateral resolution.


Valproic acid-carbapenem interaction: report of six cases and a review of the literature.
To evaluate the drug interactions between valproic acid (VPA) and carbapenem antibiotics. The effects of concurrent use of VPA and carbapenem antibiotics were evaluated in a retrospective observational study of hospitalized adults. Patients receiving both VPA and a carbapenem with at least two plasma VPA concentrations serially measured prior to, during, and/or after this combined treatment were included. Six critically ill VPA-treated patients were identified who concurrently received meropenem (n=4), imipenem (n=1), or ertapenem (n=1). As compared with values obtained while not receiving treatment with the carbapenem, mean plasma VPA trough concentrations decreased by 58% (from 51.7 [95% confidence interval {CI} 28.0-75.4] to 21.8 [95% CI 11.1-32.5] mg/L; p = 0.025). Estimated mean VPA clearance increased by 191% (from 0.0158 [95% CI 0.0041-0.0275] to 0.0302 [95% CI 0.0169-0.0591] L/h/kg; p = 0.007). All VPA concentrations measured during concurrent VPA-carbapenem treatment were below the lower boundary of the usual therapeutic range. Five patients (83%) experienced generalized seizures during concurrent VPA-carbapenem treatment, including two with no prior history of seizures or epilepsy. All recipients showed evidence of a complex pharmacokinetic and pharmacodynamic drug interaction between VPA and a carbapenem. Concurrent use of these medications should be avoided.


Characteristics of worry in GAD patients, social phobics, and controls.
Phenomenological features of worry such as thought content, subjective experience of worry, and efforts to control were investigated in the present interview study, as well as retrospective information about possible origins. To examine the clinical specificity of worrying in Generalized Anxiety Disorder (GAD), 36 GAD patients were compared to a normal control group (N = 30) and to a clinical control group (N = 22 social phobics). GAD patients differed from both groups in having higher frequency of worry, higher number of different worry topics, lower subjective controllability, more accompanying bodily symptoms, and more distress during worry. Thus, in general, our data confirm the central and specific role of worrying in GAD. Furthermore, in contrast to other topics, worrying about daily hassles was specific to GAD patients, which represents a lower threshold for starting to worry.


[Effect of 2-phenyl-3-methyl-3-hydroxypentanoic and 2,3-diphenyl-3-hydroxypentanoic acids on the lipolytic activity of rat adipose tissue].
Aromatic derivatives of mevalonic acid (2-phenyl-3-methyl-3-hydroxy pentanic and 2,3-diphenyl-3-hydroxy pentanic acids) decreased the yield of unesterified fatty acids from adipose tissue by about 28-36 percent after subcutaneous administration into rats within one or two weeks and under conditions of incubation of the adipose tissue in vitro. Distinct decrease in the yield of glycerol was cause by 2,3-diphenyl-3-hydroxy pentanic acid (1 with 10-minus 3 = 5 with 10-minus 3 M) under conditions of lipolysis stimulated by adrenaline. Antilipolytic effect of the preparation was more pronounced than that of well-known hypolipidemic drug clofibrate (p-chlorophenhydroxy isobutyrate).


Conversational Eyebrow Frowns Facilitate Question Identification: An Online Study Using Virtual Avatars.
Conversation is a time-pressured environment. Recognizing a social action (the ''speech act,'' such as a question requesting information) early is crucial in conversation to quickly understand the intended message and plan a timely response. Fast turns between interlocutors are especially relevant for responses to questions since a long gap may be meaningful by itself. Human language is multimodal, involving speech as well as visual signals from the body, including the face. But little is known about how conversational facial signals contribute to the communication of social actions. Some of the most prominent facial signals in conversation are eyebrow movements. Previous studies found links between eyebrow movements and questions, suggesting that these facial signals could contribute to the rapid recognition of questions. Therefore, we aimed to investigate whether early eyebrow movements (eyebrow frown or raise vs. no eyebrow movement) facilitate question identification. Participants were instructed to view videos of avatars where the presence of eyebrow movements accompanying questions was manipulated. Their task was to indicate whether the utterance was a question or a statement as accurately and quickly as possible. Data were collected using the online testing platform Gorilla. Results showed higher accuracies and faster response times for questions with eyebrow frowns, suggesting a facilitative role of eyebrow frowns for question identification. This means that facial signals can critically contribute to the communication of social actions in conversation by signaling social action-specific visual information and providing visual cues to speakers' intentions.


Informal and formal care among persons with dementia immediately before nursing home admission.
Dementia is a care intensive disease, especially in the later stages, implying in many cases a substantial carer burden. This study assesses the use of formal and informal care resources among persons with dementia during the last month before nursing home admission. It also describes main providers of informal care and assesses the extent of informal care rendered by the extended social network. In this cross-sectional study, we collected data about persons with dementia that were newly admitted to a nursing home in Norway. Information about the amount of formal and informal care during the last 4 weeks preceding nursing home admission was collected from the primary caregivers. Clinical data were collected by examining the patients, while sociodemographic data was collected from the patients' files. A total of 395 persons with dementia were included. The amount of informal care provided by the family caregiver was 141.9 h per month SD = 227.4. Co-resident patients received five times more informal care than non-co-residents. Informal care from the extended social network was provided to 212 patients (53.7%) with a mean of 5.6 (SD = 11.2) hours per month and represented 3.8% of the total informal care rendered to the patients. Formal care was provided to 52.7% of the patients with a mean of 18.0 (SD = 50.1) hours per month. Co-residency was significantly associated with more informal care, and the associations varied with respect to age, relation to the caregiver, and the caregiver's working situation. Good/excellent general health was associated with less formal care. Persons with dementia on the verge of admission to a nursing home are mainly supported by the family caregiver, and the use of informal care is particularly high among co-residents. In order to delay nursing home admission, future research should explore the unrealized care potential in extended social networks, as well as the potential for increasing the number of recipients of formal care services.


Electrophysiological correlates of anticipatory and poststimulus components of task switching.
Task-switching paradigms can shed light on cognitive and neural processes underlying attentional control mechanisms. An alternating runs task-switching paradigm (R. D. Rogers & S. Monsell, 1995) is used to identify ERP components associated with anticipatory and poststimulus components of task-switching processes. Subjects alternated between two tasks in a predictable series (AABB). Reaction time (RT) switch cost reduced with increasing response-stimulus (R-S) interval and a residual switch cost remained at the longest R-S interval. A switch-related positivity (D-Pos) developed in the R-S interval. D-Pos was time-locked to response onset, peaked around 400 ms post-response onset, and was unaffected by task-set interference. A switch-related negativity (D-Neg) emerged after stimulus onset. D-Neg peaked earlier with increasing R-S interval and its amplitude and latency were affected by task-set interference. D-Pos and D-Neg were interpreted within current models of task-switching.


3H-3-N-methylspiperone labels D2 dopamine receptors in basal ganglia and S2 serotonin receptors in cerebral cortex.
Detailed studies of the properties of 3H-3-N-methylspiperone (NMSP) binding in rat and human brain homogenates were performed at 37 degrees C. In homogenates of rat striatum and frontal cortex and human caudate and frontal cortex tissues, the specific binding was found to be saturable. Rat caudate contained 33.2 pmol/gm wet-weight tissue and displayed an equilibrium dissociation constant (Kd) of 8.7 X 10(-11) M; rat frontal cortex contained 18.5 pmol/gm wet-weight tissue and displayed a Kd of 1.5 X 10(-10) M. Human caudate contained 8.96 pmol/gm wet-weight tissue and displayed a Kd of 1.1 X 10(-10) M; human frontal cortex possessed 9.8 pmol/gm wet-weight tissue and a Kd of 4.4 X 10(-10) M. Kinetic studies revealed a very rapid rate of association in all the tissues studied. The rate of dissociation was relatively slow in all 4 tissue preparations; the dissociation rate was somewhat slower in rat striatum and human caudate relative to rat and human frontal cortex. This was consistent with the somewhat higher affinity, relative to frontal cortex, displayed by 3H-NMSP in rat striatal and human caudate tissue. The pharmacological properties of the specific binding in rat striatal and human caudate tissues were very similar and indicated the presence of brain D2 dopamine receptors. In rat and human frontal cortex tissue homogenates, the pharmacological characteristics of the specific binding indicated the presence of 5-HT2 receptors.


Overexpression of cyclooxygenase-2 correlates with advanced stages of colorectal cancer.
We aimed to investigate the associations of cyclooxygenase-2 (COX-2) with pathological features and survival in patients with colorectal cancer. The expression of COX-2 was examined by immunohistochemistry in 112 primary colorectal cancers, with 64 samples from the corresponding normal mucosa and 16 metastases in the regional lymph nodes of patients with colorectal cancer. The associations of COX-2 expression with clinicopathological features, including survival, were analyzed. The frequency and intensity of COX-2 staining were remarkably increased from the normal samples (17%) to the primary tumors (72%) and to the metastases (100%). Expressions of COX-2 were 25%, 74%, 78%, and 67% in Dukes' A, B, C, and D tumors, respectively (p = 0.005), and positively related to proliferative activity (p = 0.003). COX-2 expressions were 80% in colonic tumors and 60% in rectal tumors (p = 0.03). The expression of COX-2 was positively related to the better differentiated tumors in the colon (p = 0.04). We were unable to find any relationship of COX-2 with patient age, sex, tumor growth pattern, apoptosis, and patient survival (p > 0.05). We found that the expression of COX-2 was upregulated from normal cells to primary tumors and to metastases, and related to proliferative activity, tumor location, Dukes' stage, and differentiation. These results further support the evidence that COX-2 may be involved in tumorigenesis and development of colorectal cancer.


Bursting strength with various methods of renal artery ligation and potential mechanisms of failure.
Vascular control is crucial during laparoscopic ligation of the renal vessels. Intracorporeal suture ligation has been replaced by easier methods, such as specialized clip appliers and automatic stapling devices; nevertheless, the optimal application and margin of safety of such devices have yet to be determined. We sought to address this question by measuring the bursting strength of arteries ligated with several standard devices. One end of an adult porcine artery (3-7-mm diameter) was occluded with a titanium clip, self-locking polymer clip, or laparoscopic linear cutting stapler. Comparisons were made with one or two clips and with different distal cuff lengths (i.e., flush or 2 mm). The open end was secured to a pulsatile infusion pump. Leak/failure pressures were measured using a digital barometer. The mean bursting pressures for the clips were above physiologic arterial pressures (1220-1500 mm Hg). However, the vessels closed with the stapler leaked at a lower mean pressure (262 mm Hg). Failure of titanium or self-locking polymer clips was the result of vessel retraction into and behind the clip, while staple- line leakage occurred between individual staples. Bursting pressures with the titanium and self-locking polymer clips was unaffected by the number of clips or length of vascular cuff. All tested methods of vascular control performed well at physiologic pressures, suggesting that safety is not increased with traditional maneuvers such as additional clips or longer cuff length.


[Pre-operative staging of bronchial carcinomas: a comparison of computed tomography and conventional radiography (author's transl)].
Pre-operative staging was carried out in 52 out of 112 patients suspected of having bronchial carcinomas, using computed tomography and conventional radiology. CT was superior in demonstrating tumour infiltration of the mediastinum, of the thoracic wall and of metastases in mediastinal lymph nodes. Conventional radiology was better for demonstrating intrabronchial tumour and metastases of bronchopulmonary lymph nodes. By extending CT to the upper abdomen, it was possible to show metastases in 18% of patients.


CD14 is a member of the family of leucine-rich proteins and is encoded by a gene syntenic with multiple receptor genes.
We have isolated and characterized genomic and cDNA clones encoding the murine homolog of the human monocyte/granulocyte cell surface glycoprotein, CD14. As in man, the expression of murine CD14 is limited to the myeloid lineage. The murine and human CD14 genes are highly conserved in their intron-exon organization and nucleotide sequence. Their deduced protein sequences show 66% amino acid identity. In both mouse and man, the CD14 protein contains a repeating (10 times) leucine-rich motif (LXXLXLX) that is also found in a group of heterogeneous proteins from phylogenetically distant species. The CD14 gene has been mapped to mouse chromosome 18 which also contains at least five genes encoding receptors (Pdgfr, Adrb2r, li, Grl-1, Fms). Thus CD14 and the receptor genes form a conserved syntenic group localized on mouse chromosome 18 and human chromosome 5. The inclusion of CD14 in the family of leucine-rich proteins, its expression profile and the murine chromosomal localization support the hypothesis that CD14 may function as a receptor.


Treatment of impulsivity and aggression in a patient with vascular dementia.
This case illustrates how treatment with divalproex sodium, an agent approved for use as an anticonvulsant and mood stabilizer, led to a reduction in overt aggression, diminished impulsivity, and improved functional status in a patient with vascular dementia. Improved receptivity to care and reduced hostility also led to an increased quality of life for the family caregivers. The daughter also benefited greatly from participation in a family support group.


Solving satisfiability with less searching.
A new technique, complement searching, is given for reducing the amount of searching required to solve satisfiability (constraint satisfaction) problems. Search trees for these problems often contain subtrees that have approximately the same shape. When this occurs, knowledge that the first subtree does not have a solution can be used to reduce the searching in the second subtree. Only the part of the second subtree which is different from the first needs to be searched. The pure literal rule of the Davis-Putnam procedure is a special case of complement searching. The new technique greatly reduces the amount of searching required to solve conjunctive normal form predicates that contain almost pure literals (literals with a small number of occurrences).


Portable Device for Measuring Isometric Plantar-Flexion Force after Open Achilles Repair: Retrospective Cohort of 30 Recreational Athletes with One-Year Minimum Follow-Up.
Plantar flexion force in recreational athletes after repair of Achilles tendon tears has rarely been reported, due to the lack of a widely available and fast measurement method. Knowledge of this parameter would allow optimisation of muscular and sports recovery. The main objective of this study was to measure the isometric force of the triceps surae at least 1 year after unilateral Achilles-tendon repair, comparatively to the unaffected side, using a portable device. The secondary objectives were to compare Achilles tendon and calf-muscle trophicity, dorsiflexion, and the single-leg heel-rise test versus the normal side and to assess functional scores at last follow-up. (i) Plantar flexion force does not differ significantly between the operated and contralateral sides. (ii) Calf-muscle trophicity, dorsiflexion, and the single-leg heel-rise test do not differ significantly between the operated and contralateral sides. This single-centre retrospective study included patients aged 18 to 65 years with a history of open repair of an acute unilateral Achilles tear at least 1 year earlier. Plantar flexion force, calf-muscle trophicity, dorsiflexion, and the heel-to-floor distance were measured on both sides and compared. The ATRS, VISA-A score, and EFAS score were determined. All complications were recorded. The study included 30 patients with a mean follow-up of 20.3 months (range, 12-28 months). The operated side had significantly lower values for isometric triceps surae force, calf circumference, and heel-to-floor distance compared to the contralateral side (p<0.0001). Tendon width was significantly greater on the operated side (p<0.0001). Dorsiflexion did not differ significantly between sides (p=0.106). Mean functional score values were 91.5±6 for the ATRS (maximum, 100), 85±12 for the VISA-A score (maximum, 100), and 19.5±4 for the EFAS score (maximum, 24). Isometric triceps-surae force measured using a portable device at least 1 year after open surgical Achilles-tendon repair was significantly decreased compared to the contralateral side. Calf circumference and heel-to-floor distance were also significantly lower. These data indicate a need for improved monitoring of triceps surae recovery to optimise rehabilitation and sports resumption. IV, retrospective comparative cohort study.


Vaccine presentations and delivery technologies--what does the future hold?
There is an urgent need to change the presentations and delivery technologies of current vaccines. Until recently, these factors had not been key criteria in the selection of vaccines for program use. Recent and current changes in the field of vaccines and their delivery lead the authors to postulate that a major paradigm shift will take place over the next decade to revolutionize vaccine presentation and delivery in national immunization programs. The programmatic needs for certain vaccine presentations will increasingly dictate elements of vaccine development and manufacture. Over the next decade, an inexorable drift towards firstly, single-dose preparations, and secondly, delivery technologies other than the conventional needle and syringes is anticipated. A unified system capable of delivering multiple antigens as a single dose is urgently needed; however, changing the status quo of vaccine manufacture is not easy. The market predominantly produces vaccines delivered by needle and syringe. Profits for manufacturers from sales to developing countries are marginal at best, and there is little financial incentive to change. Global leaders will need to take bold decisions and begin demanding vaccines which have a presentation that lends them to safer, more practical delivery systems. If a strong enough case can be made to restructure the vaccine manufacturing industry, either through market forces, global bodies, such as the World Health Organization and the United Nations Children's Fund, or both, a dramatic change could be brought about that will make vaccine delivery simpler and safer. A globally coordinated approach to funding research and the introduction of a multiple-antigen, single-dose delivery system is urgently needed. The needs are clear, and this review argues that if the case is presented strongly enough, the resources will be found.


Resistance of South American opossums to vWF-binding venom C-type lectins.
Opossums in the clade Didelphini are well known to be resistant to snake venom due to endogenous circulating inhibitors which target metalloproteinases and phospholipases. However, the mechanisms through which these opossums cope with a variety of other damaging venom proteins are unknown. A protein involved in blood clotting (von Willebrand Factor) has been found to have undergone rapid adaptive evolution in venom-resistant opossums. This protein is a known target for a subset of snake venom C-type lectins (CTLs), which bind it and then induce it to bind platelets, causing hemostatic disruption. Several amino acid changes in vWF unique to these opossums could explain their resistance; however, experimental evidence that these changes disrupt venom CTL binding was lacking. We used platelet aggregation assays to quantify resistance to a venom-induced platelet response in two species of venom-resistant opossums (Didelphis virginiana, Didelphis aurita), and one venom-sensitive opossum (Monodelphis domestica). We found that all three species have lost nearly all their aggregation response to the venom CTLs tested. Using washed platelet assays we showed that this loss of aggregation response is not due to inhibitors in the plasma, but rather to the failure of either vWF or platelets (or both) to respond to venom CTLs. These results demonstrate the potential adaptive function of a trait previously shown to be evolving under positive selection. Surprisingly, these findings also expand the list of potentially venom tolerant species to include Monodelphis domestica and suggest that an ecological relationship between opossums and vipers may be a broader driver of adaptive evolution across South American marsupials than previously thought.


Androgen-induced programs for prostate epithelial growth and invasion arise in embryogenesis and are reactivated in cancer.
Cancer cells differentiate along specific lineages that largely determine their clinical and biologic behavior. Distinct cancer phenotypes from different cells and organs likely result from unique gene expression repertoires established in the embryo and maintained after malignant transformation. We used comprehensive gene expression analysis to examine this concept in the prostate, an organ with a tractable developmental program and a high propensity for cancer. We focused on gene expression in the murine prostate rudiment at three time points during the first 48 h of exposure to androgen, which initiates proliferation and invasion of prostate epithelial buds into surrounding urogenital sinus mesenchyme. Here, we show that androgen exposure regulates genes previously implicated in prostate carcinogenesis comprising pathways for the phosphatase and tensin homolog (PTEN), fibroblast growth factor (FGF)/mitogen-activated protein kinase (MAPK), and Wnt signaling along with cellular programs regulating such 'hallmarks' of cancer as angiogenesis, apoptosis, migration and proliferation. We found statistically significant evidence for novel androgen-induced gene regulation events that establish and/or maintain prostate cell fate. These include modulation of gene expression through microRNAs, expression of specific transcription factors, and regulation of their predicted targets. By querying public gene expression databases from other tissues, we found that rather than generally characterizing androgen exposure or epithelial budding, the early prostate development program more closely resembles the program for human prostate cancer. Most importantly, early androgen-regulated genes and functional themes associated with prostate development were highly enriched in contrasts between increasingly lethal forms of prostate cancer, confirming a 'reactivation' of embryonic pathways for proliferation and invasion in prostate cancer progression. Among the genes with the most significant links to the development and cancer, we highlight coordinate induction of the transcription factor Sox9 and suppression of the proapoptotic phospholipid-binding protein Annexin A1 that link early prostate development to early prostate carcinogenesis. These results credential early prostate development as a reliable and valid model system for the investigation of genes and pathways that drive prostate cancer.


Adventure in the outdoors. Its importance in the development of an adolescent boy.
This exploration of how the analyst works uses a 15-year-old boy's description of an adventure in the outdoors as the starting point for a sequence of associations which led first to a passage from a Wordsworth poem and then to memories from the analyst's own childhood. The related themes of rite of passage and adolescent mourning are elaborated.


The EDIT Survey: Identifying Emergency Department Information Technology Knowledge and Training Gaps.
To review Emergency Department internet connectivity, cyber risk factors, perception of risks and preparedness, security policies, training and mitigation strategies. A validated targeted survey was sent to Canadian ED physicians and nurses between March 5, 2019 and April 28, 2019. There were 349 responses, with physicians making up 84% of the respondents (59% urban teaching, 35% community teaching, 6% community non-teaching hospitals). All had multiple passwords, 93% had more than 1 user account, over 90% had to log repeatedly each workday, 52% had to change their passwords every 3 months, 75% had multiple methods of authentication and 53% reported using a terminal where someone else was already logged in. Passwords were used to review laboratory and radiology data, access medical records and manage patient flow. Majority of the respondents (51%) did not know if they worked with internet linked devices. Only 7% identified an 'air gapped' computer in their facility and 76% used personal devices for patient care, with less than a third of those allowing the IT department to review their device. A total of 26 respondents received no cyber security training. This paper revealed significant computer-human interface dysfunctionality and readiness gaps in the event of an IT failure. These stemmed from poor system design, poor planning and lack of training. The paper identified areas with technical or training solutions and suggested mitigation strategies.


Neurophysiologic exploration: a reliable tool in HIV-1 encephalopathy diagnosis in children.
HIV-1 related encephalopathy has a bad prognostic meaning in the course of AIDS disease, but the early association of different drugs can modify its course. For this reason it is very important to recognize CNS involvement as soon as possible. As shown in the literature, at least in adult studies, EEG and Evoked Potentials (EP) are good tools in evaluating CNS alterations. In children data are rare. A ten-year prospective study of 44 infected children out of 142 born from HIV-1 positive mothers has been done. The children have been submitted to EEG recording every six months in the first 18 months of life and then every year, to multimodal EP every six months. A total of 357 EEG, 47 P-VEP, 62 F-VEP and 98 BAEP have been performed. EEG: we found no pathologic results in patients belonging to category A; results were pathologic in 17.7% in category B, in 47.7% in C and in 77% of encephalopathic patients. It seems that EEG alterations are parallel to disease progression, with a relative risk of developing encephalopathy (R.R. = 1.15) and of death (R.R. = 2.33) for patients belonging to category C. We obtained a statistically significant lengthening in BAEP interpeak latency of left ear in all groups. For patients in category C the risk of developing encephalopathy is statistically significant (p = 0.045; R.R. = 6.75) and risk of death is high (R.R. = 4). Neurophysiologic exams are a reliable tool for the diagnosis of encephalopathy, in addition to clinical evidence.


Modulation of apoptosis by cancer chemopreventive agents.
A review of almost 2000 studies showed that the large majority of 39 putative cancer chemopreventive agents induced "spontaneous" apoptosis. Inhibition of the programmed cell death triggered by a variety of stimuli was consistently reported only with ascorbic acid, alpha-tocopherol, and N-acetylcysteine (NAC). We performed experimental studies in rodents exposed to cigarette smoke, either mainstream (MCS) or environmental (ECS), and UV-A/B-containing light. The nonsteroidal anti-inflammatory drug sulindac did not affect the apoptotic process in the skin of light-exposed mice and in the lungs of ECS-exposed mice. Likewise, 5,6-benzoflavone, indole-3-carbinol, 1,2-dithiole-3-thione and oltipraz failed to modulate apoptosis in the respiratory tract of ECS-exposed rats. Phenethyl isothiocyanate further enhanced the frequency of apoptosis in pulmonary alveolar macrophages and bronchial epithelial cells, and upregulated several genes in the lung of ECS-exposed rats. Both individually and in combination with oltipraz, NAC inhibited apoptosis in the respiratory tract of rats exposed either to MCS or ECS. Moreover, NAC attenuated the ECS-related overexpression of proapoptotic genes and normalized the levels of proapoptotic proteins in rat lung. The transplacental administration of NAC to mice considerably attenuated gene overexpression in the liver of fetuses exposed to ECS throughout pregnancy. Inhibition of apoptosis by chemopreventive agents reflects their ability to counteract certain upstream signals, such as genotoxic damage, redox imbalances, and other forms of cellular stress that trigger apoptosis. On the other hand, enhancement of apoptosis is a double-edged sword, since it represents a protective mechanism in carcinogenesis but may contribute to the pathogenesis of other degenerative diseases. We suggest that stimulation of apoptosis by so many chemopreventive agents, as reported in the literature, may often reflect the occurrence of toxic effects at high doses.


Chaos synchronization communication using extremely unsymmetrical bidirectional injections.
Chaos synchronization and message transmission between two semiconductor lasers with extremely unsymmetrical bidirectional injections (EUBIs) are discussed. By using EUBIs, synchronization is realized through injection locking. Numerical results show that if the laser subjected to strong injection serves as the receiver, chaos pass filtering (CPF) of the system is similar to that of unidirectional coupled systems. Moreover, if the other laser serves as the receiver, a stronger CPF can be obtained. Finally, we demonstrate that messages can be extracted successfully from either of the two transmission directions of the system.


Segmental expression of the EphA4 (Sek-1) receptor tyrosine kinase in the hindbrain is under direct transcriptional control of Krox-20.
Segmentation of the vertebrate hindbrain leads to the formation of a series of rhombomeres (r) with distinct identities. Recent studies have uncovered regulatory links between transcription factors governing this process, but little is known of how these relate to molecules mediating cell-cell signalling. The Eph receptor tyrosine kinase gene EphA4 (Sek-1) is expressed in r3 and r5, and function-blocking experiments suggest that it is involved in restricting intermingling of cells between odd- and even-numbered rhombomeres. We have analysed the cis-acting regulatory sequences of the EphA4 gene in transgenic mice and identified a 470 bp enhancer element that drives specific expression in r3 and r5. Within this element, we have identified eight binding sites for the Krox-20 transcription factor that is also expressed in r3 and r5. Mutation of these binding sites abolishes r3/r5 enhancer activity and ectopic expression of Krox-20 leads to ectopic activation of the enhancer. These data indicate that Krox-20 is a direct transcriptional activator of EphA4. Together with evidence that Krox-20 regulates Hox gene expression, our findings reveal a mechanism by which the identity and movement of cells are coupled such that sharply restricted segmental domains are generated.


Changes in behavior and salivary cortisol after targeted cognitive training in typical 12-month-old infants.
Previous research has suggested that early development may be an optimal period to implement cognitive training interventions, particularly those relating to attention control, a basic ability that is essential for the development of other cognitive skills. In the present study, we administered gaze-contingent training (95 min across 2 weeks) targeted at voluntary attention control to a cohort of typical 12-month-old children (N = 24) and sham training to a control group (N = 24). We assessed training effects on (a) tasks involving nontrained aspects of attention control: visual sustained attention, habituation speed, visual recognition memory, sequence learning, and reversal learning; (b) general attentiveness (on-task behaviors during testing); and (c) salivary cortisol levels. Assessments were administered immediately after the cessation of training and at a 6-week follow-up. On the immediate posttest infants showed significantly more sustained visual attention, faster habituation, and improved sequence learning. Significant effects were also found for increased general attentiveness and decreased salivary cortisol. Some of these effects were still evident at the 6-week follow-up (significantly improved sequence learning and marginally improved sustained attention). These findings extend the emerging literature showing that attention training is possible in infancy. (PsycINFO Database Record


Pulmonary Hypertension-Associated Right Ventricular Cardiomyocyte Remodelling Reduces Treprostinil Function.
(1) Pulmonary hypertension (PH)-associated right ventricular (RV) failure is linked to a reduction in pulmonary vasodilators. Treprostinil has shown effectiveness in PAH patients with cardiac decompensation, hinting at potential cardiac benefits. We investigated treprostinil's synergy with isoprenaline in RV and LV cardiomyocytes. We hypothesised that disease-related RV structural changes in cardiomyocytes would reduce contractile responses and cAMP/PKA signalling activity. (2) We induced PH in male Sprague Dawley rats using monocrotaline and isolated their ventricular cardiomyocytes. The effect of in vitro treprostinil and isoprenaline stimulation on contraction was assessed. FRET microscopy was used to study PKA activity associated with treprostinil stimulation in AKAR3-NES FRET-based biosensor-expressing cells. (3) RV cells exhibited maladaptive remodelling with hypertrophy, impaired contractility, and calcium transients compared to control and LV cardiomyocytes. Combining treprostinil and isoprenaline failed to enhance inotropy in PH RV cardiomyocytes. PH RV cardiomyocytes displayed an aberrant contractile behaviour, which the combination treatment could not rectify. Finally, we observed decreased PKA activity in treprostinil-treated PH RV cardiomyocytes. (4) PH-associated RV cardiomyocyte remodelling reduced treprostinil sensitivity, inotropic support, and impaired relaxation. Overall, this study highlights the complexity of RV dysfunction in advanced PH and suggests the need for alternative therapeutic strategies.


Effect and mechanism of nociceptin/orphanin FQ reversing multi-drug resistance in K562/ADM cell.
To investigate the effect and mechanism of nociceptin/orphanin FQ (OFQ) reversing multi-drug resistance of K562/ADM cells in vitro. MTT assay, Wright staining, flow cytometry, transmission electron microscope and gel electrophoresis were used to evaluate the effect and mechanism of OFQ in reversing multi-drug resistance of K562/ADM cells. OFQ could time-dependently reverse the ADM resistance of K562/ADM cell. After treatment with OFQ (1 x 10(-7) mol x L(-1)), K562/ADM cells were cultured for 24, 48 and 72 h. The reversal index (RI) was 1.33, 1.42 and 1.53, respectively. Furthermore, OFQ significantly increased the intracellular accumulation of ADM in K562/ADM cells and percentage apoptosis in K562/ADM cells. OFQ down-regulated the level of P-gp time-dependently, while the level of Fas and FasL were up-regulated. There were evidently significant differences compared with the control (P < 0.01). After treating K562/ADM cells with OFQ (1 x 10(-7) mol x L(-1)) and ADM (20 microg x ml(-1)) for 48 hours, the cells showed apoptotic nuclear fragmentation, which was characterized by the appearance of a DNA ladder pattern in genomic DNA gel electrophoresis. OFQ can reverse the ADM resistance of K562/ADM cells. The mechanism involves OFQ up-regulating the expression of Fas/FasL, down-regulating the level of P-gp, and decreasing the intracellular level of calcium in K562/ADM cells.


Analysis of 1-aminopyrene-3,6,8-trisulfonate-derivatized oligosaccharides by capillary electrophoresis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), has been used for structure characterization of 1-aminopyrene-3,6,8-trisulfonate (APTS)-derivatized oligosaccharides previously separated by capillary electrophoresis (CE). The resolved components were first isolated by employing an automated high-resolution fraction collector. Using on-probe sample cleanup with a cation-exchange resin and a matrix mixture of (1:1) 6-hydroxypicolinic acid and 3-hydroxypicolinic acid, APTS-labeled oligosaccharides were successfully detected by MALDI-TOF MS in the negative ionization mode. Each APTS-labeled oligosaccharide produced single [M-H]-1 peaks. Detection limits for standard APTS-derivatized oligosaccharide (maltoheptaose) were down to 30 fmol. APTS labeled maltooligosaccharides and various standard carbohydrates were separated and collected by CE, followed by molecular weight determination with MALDI-TOF MS. The mass spectral characterization enhances the power of the CE analysis of oligosaccharides.


[Influence of atropine on the chronotropic effects of stimulating the parasympathetic innervation of the heart].
Changes of the positive chronotropic effect in the frog atrium during stimulation of the extracardiac pathways or of the central end of intracardiac innervation after application of atropin (10(-4)--10(-5) on the sinus node, were studied. The positive chronotropic effects were abolished by the atropin application. The postganglionic impulse activity of the intracardiac nervous pathways did not change. Atropin had no influence on the sympathetic acceleration of the heart rate due to stimulation of sympathetic pathways. Positive chronotropic effects induced by stimulation of the parasympathetic extra or intracardiac pathways, are of cholinergic nature.


Yolk proteolysis in rainbow trout oocytes after serum-free culture: evidence for a novel biochemical mechanism of atresia in oviparous vertebrates.
Our recent studies show little evidence for increased granulosa cell apoptosis during atresia in teleost follicles, in direct contrast to the mammalian model. Histological evidence suggests that atresia in many oviparous vertebrates involves proteolytic degradation of the energy-rich yolk storage proteins within the oocyte. This study tests the hypothesis that physiological conditions that promote atresia (hormone withdrawal) lead to increased lysosomal protease activity in rainbow trout oocytes. We subjected rainbow trout ovarian follicles to conditions that promote atresia (serum-free culture) for up to 72 hr, and measured the activity of lysosomal proteases using routine enzymatic assays. Furthermore, we used high performance liquid chromatography to quantify the increase in free amino acids resulting from proteolysis of yolk proteins. Concomitantly, we evaluated the extent of follicular apoptosis during prolonged serum-free culture, using caspase-3-like activity and DNA fragmentation as indicators of apoptosis. Our results show a significant, time-dependent increase in cathepsin L-like, but not cathepsin D-like, activity levels during culture in serum-free medium; increased cathepsin L-like activity is confirmed by a significant increase in oocyte free amino acid content after 72 hr culture. In contrast, we detected only a transient increase in apoptosis during prolonged serum-free culture, as revealed through both radioactive 3'end-labeling of oligonucleosomal DNA fragments, and caspase-3-like activity. The results of this study provide the first evidence for a novel mechanism of follicular atresia in teleosts involving cathepsin-mediated yolk proteolysis.


Ecophysiology of polyphosphate-accumulating organisms and glycogen-accumulating organisms in a continuously aerated enhanced biological phosphorus removal process.
To investigate the ecophysiology of populations of polyphosphate-accumulating organisms (PAO) and glycogen-accumulating organisms (GAO) in communities of a novel acetate fed process removing phosphate from wastewater. Attempts were made to see if acetate could be replaced by an alternative carbon source which did not support the growth of the GAO. A continuously aerated sequencing batch reactor was operated with different acetate feed levels. Fluorescence in situ hybridization (FISH) showed that Defluviicoccus GAO numbers increased at lower acetate feed levels. With FISH/microautoradiography (MAR) both detected morphotypes of Defluviicoccus assimilated a wider range of substrates aerobically than Accumulibacter PAO. Their uptake profile differed from that reported for the same phylotype in full scale anaerobic : aerobic EBPR plants. This suggests that replacing acetate with another substrate is unlikely to provide Accumulibacter with a selective advantage in this process. Why Defluviicoccus appeared to out-compete Accumulibacter at lower acetate concentrations was not clear. Data suggest physiological and morphological diversity may exist within a single Defluviicoccus phylotype. This study implies that the current FISH probes for Defluviicoccus GAO may not reveal the full extent of their biodiversity, and that more information is required before strategies for their control can be devised.