Trephination of donor corneal buttons: a scanning electron microscopic study.
Corneal buttons were cut from cat and human donor eyes by anterior trephination (with or without scissor completion) and from excised corneoscleral rims by posterior trephination (with or without a press). Scanning electron microscopy was used to evaluate the slope and quality of the cut edge, the diameter of the section of Descemet's membrane that was actually cut, and the loss of endothelial cells around the cut edge. With anterior trephination, the diameter of the section of Descemet's membrane that is cut was found to be less than that of the trephine because the cut edge slopes inward. Posterior trephination with or without a press consistently produced circular buttons with accurately cut outward-sloping edges. This resulted in a diameter of Descemet's membrane which was greater than in the anteriorly trephinated buttons. There also was less damage to peripheral Descemet's membrane and endothelium with the posterior method, especially when compared to scissor cut buttons. A 360 degree peripheral ring of Descemet's membrnae (0.05 to 0.28 mm in width) was denuded of endothelial cells with all techniques.


Detailed characterization of proanthocyanidins in skin, seeds, and wine of Shiraz and Cabernet Sauvignon wine grapes (Vitis vinifera).
The distribution of proanthocyanidin (PA) polymer lengths, proanthocyanidin concentration at each polymer length, and polymer composition were determined in the seed, skin, and wine of Shiraz and Cabernet Sauvignon grape berries grown in southeast Australia. PA was fractionated by semipreparative high performance liquid chromatography (HPLC) and analyzed by phloroglucinolysis and HPLC to report the degree of polymerization (DP), concentration, and composition at 11 DP values in seed and wine and 21 DP values in skin. In skin, the highest PA concentration was observed at a DP of 31 in Shiraz and 29 in Cabernet Sauvignon representing 15% of the total PA in both varieties. The distribution of seed PA had the highest concentration at a DP of 7 in Shiraz and 6 in Cabernet Sauvignon representing around 30% of the total PA. In the wine PA distribution, the highest concentration was observed at a DP of 11 in Shiraz and 9 in Cabernet Sauvignon representing around 26 and 32% of the distribution, respectively. A second peak in wine PA concentration was observed at the largest DP of 18 in Shiraz and 15 in Cabernet Sauvignon representing around 20% of the distribution. The composition in wine did not vary at different DP, but the proportion of epicatechin gallate varied in seed PA less than 4 DP. The proportion of epigallocatechin increased with increasing DP in skin PA. Wine PA had a DP range and composition similar to the distribution of skin PA between DP 4 and 18 suggesting that larger skin PAs are not extracted into wine. This study provides information that could be used to target the important PA fractions in grapes that need to be measured to understand (or predict) PA extraction into wine and eventual mouthfeel.


Effects of triiodothyronine and propylthiouracil on regeneration of catecholaminergic nerve terminals in the paraventricular hypothalamic nucleus of the adult rat.
Effects of triiodothyronine and the antithyroid drug, propylthiouracil, on regeneration of catecholaminergic nerve terminals in the paraventricular hypothalamic nucleus of adult rats were studied. Lesions were produced by 6-hydroxydopamine neurotoxin and then the animals were treated with triiodothyronine or propylthiouracil inducing hyperthyroidism or hypothyroidism, respectively, as determined by radioimmunoassay. Although catecholaminergic varicosities increased with time in the paraventricular hypothalamic nucleus of rats after lesion, fluorescent microscopic quantitation showed no statistical difference in their number between rats treated with triiodothyronine and the vehicle for as long as 56 days. Furthermore, electron microscopic quantitation at 56 days postlesion showed no significant difference between the triiodothyronine-treated and control rats in terms of the density, proportion, size, types of synapses, and synaptic frequency of catecholaminergic nerve terminals. There were growth cones in the paraventricular hypothalamic nucleus, suggesting growth activity after lesion. However, we found that exogenous administration of large doses of triiodothyronine at 25 micrograms/kg had little effect on the enhancement of regeneration of central catecholaminergic terminals after their destruction by 6-hydroxydopamine.


Phospholipid transfer protein in hemodialysis patients.
Phospholipid transfer protein (PLTP) is mainly involved in high-density lipoprotein (HDL) metabolism. The role of PLTP in atherogenesis is still controversial. We aimed to investigate PLTP activity in hemodialysis (HD) patients, a population which has an increased risk for the development of atherosclerosis. PLTP activity and other markers were analyzed in blood samples from 68 HD patients and in a matched group of 68 healthy controls. Serum PLTP activity was nearly doubled in HD patients in comparison to healthy controls (median 43.0 vs. 22.4 pmol/mul/h, p < 0.001). In HD patients, PLTP activity correlated with HDL-C (r = 0.342, p = 0.004), but not with CRP (r = -0.057, p = 0.644) or leukocyte count (r = 0.116, p = 0.345). After a follow-up of 2 years, 26 HD patients had died. Kaplan-Meier analyses showed that low CRP (p = 0.047) but neither high HDL-C (p = 0.071) nor low PLTP activity (p = 0.853) were relevantly related to survival of HD patients. An elevated PLTP activity in HD patients may be considered as a further aspect of uremic dyslipidemia in HD patients. However, PLTP activity was not related to markers of inflammation or to survival of HD patients, even though it correlated with HDL-C. Thus, we conclude that PLTP does not influence the prognostically relevant inflammatory process in HD patients although it does influence the composition of HDL particles.


Anomaly of the femoral artery passage through the substance of sartorius muscle. Clinical consequences.
The passage of the femoral artery through the substance of the sartorius muscle and its clinical consequences are presented. In the series of cadavers studied, the incidence of occurrence of this condition is probably less than 0.001% in both Caucasians and black Africans. It is suggested that this anomaly which rarely occurs is due to the angioblastic column of cells forming the femoral artery passing through the myoblastic column of cells forming the sartorius muscle.


[Circulatory disorders of the retina].
Cardiovascular diseases lead to systemic vascular regression, which, among other consequences, may result in an impairment of retinal perfusion. In view of the high level of comorbidity observed in patients with circulatory problems affecting the retina, the early recognition of mild forms of vascular retinopathy is of considerable importance. A major prerequisite for an improvement in outcome is the recognition of the fact that most circulatory disorders of the retina are due to systemic vascular disease, and the eye is an important prognostic indicator of cardiovascular morbidity and mortality. As a result, effective communication between family physician/internist and ophthalmologist is basic to the successful treatment of these diseases of the eye.


Steroid excretion during the ovarian cycle in captive and wild muriquis, Brachyteles arachnoides.
Urine, feces, and copulation frequency were collected from two captive muriqui females, Brachyteles arachnoides, at the Centro de Primatologia do Rio de Janeiro following the resumption of postpartum ovarian cycles. Fecal steroid profiles from seven wild muriqui females at the Estação Biologica de Caratinga, Minas Gerais, Brazil, were compared to the captive females to determine the approximate patterns of steroid excretion relative to the urinary LH peak. Hormonal profiles from one of the captive female muriquis revealed a discrete urinary LH peak. For this female, fecal progesterone increased on the same day as the urinary LH peak, while fecal estradiol increased 6 days later and urinary steroids increased 5 days later. For both captive females, the onset of fecal progesterone increase was preceded by the onset of copulations, which occurred during at least a 5-day period. The complete fecal hormonal profiles of the one captive female for which continuos data were available were similar to those found in wild muriqui monkeys, with the onset of an increase in sustained progesterone levels occurring several days prior to the onset of sustained estradiol increase. These patterns suggest that fecal progesterone may be excreted rapidly in this species. The onset of sustained increase in fecal progesterone levels, together with the consistent delay in the onset of the sustained increase in estradiol, may provide the best indicators of the periovulatory period for muriqui females.


A minimal 3D model of mosquito flight behaviour around the human baited bed net.
Advances in digitized video-tracking and behavioural analysis have enabled accurate recording and quantification of mosquito flight and host-seeking behaviours, facilitating development of individual (agent) based models at much finer spatial scales than previously possible. Quantified behavioural parameters were used to create a novel virtual testing model, capable of accurately simulating indoor flight behaviour by a virtual population of host-seeking mosquitoes as they interact with and respond to simulated stimuli from a human-occupied bed net. The model is described, including base mosquito behaviour, state transitions, environmental representation and host stimulus representation. In the absence of a bed net and human host bait, flight distribution of the model population was relatively uniform throughout the arena. Introducing an unbaited untreated bed net induced a change in distribution with an increase in landing events on the net surface, predominantly on the sides of the net. Adding the presence of a simulated human bait dramatically impacted flight distribution patterns, exploratory foraging and, the number and distribution of landing positions on the net, which were determined largely by the orientation of the human within. The model replicates experimental results with free-flying living mosquitoes at human-occupied bed nets, where contact occurs predominantly on the top surface of the net. This accuracy is important as it quantifies exposure to the lethal insecticide residues that may be unique to the net roof (or theoretically any other surface). Number of net contacts and height of contacts decreased with increasing attractant dispersal noise. Results generated by the model are an accurate representation of actual mosquito behaviour recorded at and around a human-occupied bed net in untreated and insecticide-treated nets. This fine-grained model is highly flexible and has significant potential for in silico screening of novel bed net designs, potentially reducing time and cost and accelerating the deployment of new and more effective tools for protecting against malaria in sub-Saharan Africa.


Time course and interrelationship of dysrhythmias in patients with surgically repaired atrial septal defect.
Atrial fibrillation (AF) and other supraventricular tachycardias (SVTs) are known complications after surgical repair of atrial septal defect (ASD), but sinus node dysfunction (SND) and complete atrioventricular conduction block (cAVB) may also occur. The aim of this study was to examine time course and interrelationship of various dysrhythmias in patients with ASD. Adult patients (N = 95) with surgically repaired secundum ASD (n = 40), partial atrioventricular septal defect (n = 37) or sinus venosus defect (n = 18), and documented SND, cAVB, AF, and/or other SVT were included. The median age at repair was 13 years (interquartile range [IQR] 6-45 years), and patients were followed for 26 years (IQR 15-37 years) after ASD repair. SND was observed in 34 patients (36%), cAVB in 14 (14%), AF in 48 (49%), and SVT in 44 (45%); 37 patients (39%) had ≥2 dysrhythmias. All dysrhythmias presented most often after ASD repair (P < .01), with a median duration of 12 years (IQR 17 days - 32 years) to 16 years (IQR 4 - 28 years) between repair and onset. Development of SND and cAVB late after ASD repair was not related to a redo procedure in 100% and 60% of patients, respectively. SND preceded atrial tachyarrhythmias in 50% (P = .31) and SVT preceded AF in 68% (P = .09) of patients with both dysrhythmias. A substantial number of dysrhythmias presented (very) late after ASD repair. In most patients, development of late SND and cAVB was not related to redo procedures. In patients with multiple dysrhythmias, a specific order of appearance was not observed.


Review and comparison of current trends in the postoperative management of tendon repair.
The precision of the Evans/Burkhalter protocol and the work by Silverman and associates exemplify one of the most valuable of all current trends in rehabilitation of the healing tendon. Knowledge of tendon excursion at each level and throughout the range of motion in each joint gives us safe parameters for tendon mobilization. Hand rehabilitation is becoming more of a science while remaining an art. Research into tendon healing, nutrition, anatomy, biomechanics, and physiology gives us a solid basis for our treatment techniques. We now need to replicate studies already performed and quantify more precisely the data we have. Many questions remain unanswered. There is a wide variety in the position of splinting for flexor tendon mobilization under current protocols: What joint positions are optimal and why? The number and frequency of repetitions in early mobilization protocols varies greatly: What number and frequency is more appropriate for which patients? How much tendon excursion will control adhesions, promote healing, and avoid gap formation or elongation of the repair? How much force should we apply passively to maintain or increase joint motion? How soon should we start active motion, and how can we control the strength of those early muscle contractions? Do "place-hold" exercises truly place less tension on the repair site? How soon should we begin resisted exercise, and how much resistance are we applying with each type of exercise? Should blocking exercises be considered resistive? How should tendon management protocols be adapted in the presence of associated injuries? Lack of space has prevented discussion here of recent and needed research in a number of areas, such as the effectiveness and appropriate precautions for the use of ultrasound, iontophoresis, and neuromuscular electrical stimulation in tendon management. The evidence is growing, but we have a long way to go. To improve our clinical results, the trend toward precision must continue and grow.


A life-breaking event: early experiences of the consequences of a hip fracture for elderly people.
To explore and describe the consequences of an acute hip fracture as experienced by home-dwelling elderly people shortly after discharge from hospital. Semi-structured interviews using the phenomenographic method. Eighteen subjects were interviewed in their own homes one month after discharge. The interviewees described experiences of changes in their relation to the body, themselves, to others and to their whole life situation. These experiences were described as being limited in movement, having lost confidence in the body, becoming humble and grateful, respecting oneself and one's own needs, becoming more dependent on others, gaining more human contact and being treated in a friendly way by others, being secluded and trapped at home, feeling old, closer to death and having lost their zest for life, and taking one day at a time and being uncertain about the future. Experienced consequences of a hip fracture were multidimensional and involved dramatic changes in the interviewees' life situation, including existential thoughts and reappraisal of the years of life that remained. The results indicate that the fracture seemed not only to break the bone but also to cause social and existential cracks, as experienced in the early phase after the injury.


Ectopic positioning of pacemaker electrode.
The present case reports on a patient presenting the cardiac form of Chagas disease, with left ventricular dysfunction and second-degree atrioventricular block Mobitz type II, associated with several syncope episodes. The patient underwent a double-chamber definitive artificial pacemaker implant. One year after the implant, the displacement of the atrial electrode was diagnosed and the patient was submitted to re-implantation of the atrial electrode. Two years after the first surgical procedure, the patient presented dyspnea on exertion. The physical evaluation included an echocardiogram, which detected the presence of a foreign body with metallic characteristics in the left cardiac chambers, consistent with that of an ectopic pacemaker electrode.


Post-Transcriptional Control in the Regulation of Polyhydroxyalkanoates Synthesis.
The large production of non-degradable petrol-based plastics has become a major global issue due to its environmental pollution. Biopolymers produced by microorganisms such as polyhydroxyalkanoates (PHAs) are gaining potential as a sustainable alternative, but the high cost associated with their industrial production has been a limiting factor. Post-transcriptional regulation is a key step to control gene expression in changing environments and has been reported to play a major role in numerous cellular processes. However, limited reports are available concerning the regulation of PHA accumulation in bacteria, and many essential regulatory factors still need to be identified. Here, we review studies where the synthesis of PHA has been reported to be regulated at the post-transcriptional level, and we analyze the RNA-mediated networks involved. Finally, we discuss the forthcoming research on riboregulation, synthetic, and metabolic engineering which could lead to improved strategies for PHAs synthesis in industrial production, thereby reducing the costs currently associated with this procedure.


Surface thermodynamics of bacterial adhesion.
The adhesion of five strains of bacteria, i.e., Staphylococcus aureus (strain 049), Staphylococcus epidermidis (strain 047), Escherichia coli (strains 055 and 2627), and Listeria monocytogenes, to various polymeric surfaces was studied. The design of the experimental protocol was dictated by thermodynamic considerations. From the thermodynamic model for the adhesion of small particles from a suspension onto a solid substratum, it follows that the extent of adhesion is determined by the surface properties of all three phases involved, i.e., the surface tensions of the adhering particles, of the substrate, and of the suspending liquid medium. In essence, adhesion is more extensive to hydrophilic substrata (i.e., substrata of relatively high surface tension) than to hydrophobic substrata, when the surface tension of the bacteria is larger than that of the suspending medium. When the surface tension of the suspending liquid is larger than that of the bacteria, the opposite pattern of behavior prevails. Suspensions of bacteria at a concentration of 10(8) microorganisms per ml were brought into contact with several polymeric surfaces (Teflon, polyethylene, polystyrene, and acetal and sulfonated polystyrene) for 30 min at 20 degrees C. After rinsing, the number of bacteria adhering per unit surface area was determined by image analysis. The surface tension of the suspending medium. Hanks balanced salt solution, was modified through the addition of various amounts of dimethyl sulfoxide. It was found that the number of bacteria adhering per unit surface area correlates well with the thermodynamic predictions and that these data may be used to determine the surface tension of the different bacterial species. The surface tensions of the bacteria obtained in this fashion are in excellent agreement with those obtained by other methods.


Different mechanisms of serum complement activation in the plasma of common (Chelydra serpentina) and alligator (Macrochelys temminckii) snapping turtles.
Reptiles are declining worldwide yet our understanding of their immune function lags far behind other taxa. The innate immune system is the primary mode of defense in reptiles, and the serum complement cascade is its major component. We assessed serum complement activity of plasma in two closely related aquatic turtle species, the common snapping turtle (CST; Chelydra serpentina) and alligator snapping turtle (AST; Macrochelys temminckii). We used a sheep red blood cell (SRBC) hemolysis assay to assess serum complement activity. Although the antibacterial activities of the plasma of these turtle species are similar, the hemolytic activity was much stronger in CST than AST. Treatment with inhibitors of the serum complement cascade indicated differences in the mechanisms of complement activation between the turtle species. We subjected plasma from both turtle species to mannan affinity chromatography and analyzed the eluate with SDS-PAGE, which revealed that plasma from the CSTs contained only small amounts of one C-type lectin protein while the AST plasma contained high concentrations of two C-type lectins (31.0 and 35.9 kDa). Edman degradation analyses confirmed that the two AST proteins contained identical N-terminal sequences. Thus, the CST appears to rely more heavily on the alternative mechanism of serum complement activation, while the AST appears to rely more on the lectin-mediated pathway, which is a pattern recognition response to prokaryotes not activated by the SRBCs. These results are unique in that the use of serum complement pathways are generally assumed to be conserved within clades.


Effect of platelet-rich plasma on peripheral nerve regeneration.
Activated platelets release various growth factors, some of which are recognized to improve nerve regeneration. This study evaluated the effect of platelet-rich plasma (PRP) in end-to-end neurorrhaphy. A total of 45 Wistar rats were used, with the initial five used for PRP preparation. The right hind limbs were used as experimental, with the left as control. The animals were treated in five groups. Group A (n = 4): The right sciatic nerve was dissected only from the sciatic notch to the bifurcation. In all other groups, the nerve was sharply transected and repaired with: group B (n = 8): two sutures; group C (n = 8): six sutures; group D (n = 10): two sutures and PRP; and group E (n = 10): six sutures and PRP. Groups D and E were compared with groups B and C, respectively. Group E had a shorter latency time in electromyography ( P < 0.01) and a thicker myelin layer in the histological evaluation ( P < 0.003) in comparison with group C. These positive effects of PRP were not detected in the nerves were repaired with two sutures. In this animal model, the application of PRP to the repair site helped to improve remyelinization of the sciatic nerve in rats when the epineural repair was done with six sutures.


Compensating time delays with neural predictions: are predictions sensory or motor?
Neural delays are a general property of computations carried out by neural circuits. Delays are a natural consequence of temporal summation and coding used by the nervous system to integrate information from multiple resources. For adaptive behaviour, however, these delays must be compensated. In order to sense and interact with moving objects, for example, the visual system must predict the future position of the object to compensate for delays. In this paper, we address two critical questions concerning the implementation of the compensation mechanisms in the brain, namely, where does compensation occur and how is it realized. We present evidence showing that compensation can happen in both the motor and sensory systems, and that compensation using 'diagonal neural pathways' is a suitable strategy for implementing compensation in the visual system. In this strategy, neural signals in the early stage of information processing are sent to the future cortical positions that correspond to the distance the object will travel in the period of transmission delay. We propose a computational model to elucidate this using the retinal visual information pathway.


Autoimmune diabetes onset results from qualitative rather than quantitative age-dependent changes in pathogenic T-cells.
Diabetogenic T-cells can be detected in pre-diabetic nonobese diabetic (NOD) mice after transfer in NOD-SCID recipients. Here we demonstrate that 6-week-old pre-diabetic NOD mice, >2 months before disease onset, already harbor pathogenic T-cells in equal numbers to overtly diabetic animals. The delay in diabetes appearance is explained by the presence of regulatory CD4+ CD25+ T-cells that control diabetogenic effectors and that are, in our hands, transforming growth factor (TGF)-beta-dependent. Our present results suggest, however, that diabetes onset is only partly explained by a decline in this regulatory T-cell activity. Another major factor appears to be the progressive resistance of diabetogenic cells to TGF-beta-dependent mediated inhibition. We propose that progression to overt disease correlates with the pathogenic T-cell's escape from TGF-beta-dependent T-cell-mediated regulation.


Randomized study on the efficacy of cognitive-behavioral therapy for insomnia secondary to breast cancer, part I: Sleep and psychological effects.
Chronic insomnia is highly prevalent in cancer patients. Cognitive-behavioral therapy (CBT) is considered the treatment of choice for chronic primary insomnia. However, no randomized controlled study has been conducted on its efficacy for insomnia secondary to cancer. Using a randomized controlled design, this study conducted among breast cancer survivors evaluated the effect of CBT on sleep, assessed both subjectively and objectively, and on hypnotic medication use, psychological distress, and quality of life. Fifty-seven women with insomnia caused or aggravated by breast cancer were randomly assigned to CBT (n = 27) or a waiting-list control condition (n = 30). The treatment consisted of eight weekly sessions administered in a group and combined the use of stimulus control, sleep restriction, cognitive therapy, sleep hygiene, and fatigue management. Follow-up evaluations were carried out 3, 6, and 12 months after the treatment. Participants who received the insomnia treatment had significantly better subjective sleep indices (daily sleep diary, Insomnia Severity Index), a lower frequency of medicated nights, lower levels of depression and anxiety, and greater global quality of life at post-treatment compared with participants of the control group after their waiting period. Results were more equivocal on polysomnographic indices. Therapeutic effects were well maintained up to 12 months after the intervention and generally were clinically significant. This study supports the efficacy of CBT for insomnia secondary to breast cancer.


Radiolabeling of Mycobacterium leprae lipids within schwannoma cells, a potential drug screening system.
This study describes a novel method which could be developed into a test system of evaluating the efficacy of antileprosy drugs. The method estimates incorporation of [14C]acetate into lipids of Mycobacterium leprae maintained within the 33B Schwannoma cell line. Schwannoma cell-resident M. leprae cells incorporated significant levels of radiolabel within their lipids during 12 days of incubation in vitro. This incorporation was markedly reduced by 5 micrograms of rifampin per ml (decrease, 81.62%); this decrease was observed within 24 h of addition of the drug. Dapsone also reduced the radiolabel incorporation into the lipids, but to a lesser extent (decrease, 27.58%). This system was also able to differentiate between rifampin-sensitive and -resistant strains of mycobacteria. It is suggested that since the effect of bacteriostatic (dapsone) and bactericidal (rifampin) drugs could be detected by using this technique, it may prove useful in screening novel drugs acting against M. leprae.


Identifying Severe Stroke Patients Likely to Benefit From Thrombectomy Despite Delays of up to a Day.
Selected patients with large vessel occlusions (LVO) can benefit from thrombectomy up to 24 hours after onset. Identifying patients who might benefit from late intervention after transfer from community hospitals to thrombectomy-capable centers would be valuable. We searched for presentation biomarkers to identify such patients. Frequent MR imaging over 2 days of 38 untreated LVO patients revealed logarithmic growth of the ischemic infarct core. In 24 patients with terminal internal carotid artery or the proximal middle cerebral artery occlusions we found that an infarct core growth rate (IGR) <4.1 ml/hr and initial infarct core volumes (ICV) <19.9 ml had accuracies >89% for identifying patients who would still have a core of <50 ml 24 hours after stroke onset, a core size that should predict favorable outcomes with thrombectomy. Published reports indicate that up to half of all LVO stroke patients have an IGR <4.1 ml/hr. Other potentially useful biomarkers include the NIHSS and the perfusion measurements MTT and Tmax. We conclude that many LVO patients have a stroke physiology that is favorable for late intervention, and that there are biomarkers that can accurately identify them at early time points as suitable for transfer for intervention.


Pharmacological characterization of AZD5069, a slowly reversible CXC chemokine receptor 2 antagonist.
In normal physiologic responses to injury and infection, inflammatory cells enter tissue and sites of inflammation through a chemotactic process regulated by several families of proteins, including inflammatory chemokines, a family of small inducible cytokines. In neutrophils, chemokines chemokine (CXC motif) ligand 1 (CXCL1) and CXCL8 are potent chemoattractants and activate G protein-coupled receptors CXC chemokine receptor 1 (CXCR1) and CXCR2. Several small-molecule antagonists of CXCR2 have been developed to inhibit the inflammatory responses mediated by this receptor. Here, we present the data describing the pharmacology of AZD5069 [N-(2-(2,3-difluorobenzylthio)-6-((2R,3S)-3,4-dihydroxybutan-2-yloxy)[2,4,5,6-(13)C4, 1,3-(15)N2]pyrimidin-4-yl)azetidine-1-sulfonamide,[(15)N2,(13)C4]N-(2-(2,3-difluoro-6-[3H]-benzylthio)-6-((2R,3S)-3,4-dihydroxybutan-2-yloxy)pyrimidin-4-yl)azetidine-1-sulfonamide], a novel antagonist of CXCR2. AZD5069 was shown to inhibit binding of radiolabeled CXCL8 to human CXCR2 with a pIC50 value of 9.1. Furthermore, AZD5069 inhibited neutrophil chemotaxis, with a pA2 of approximately 9.6, and adhesion molecule expression, with a pA2 of 6.9, in response to CXCL1. AZD5069 was a slowly reversible antagonist of CXCR2 with effects of time and temperature evident on the pharmacology and binding kinetics. With short incubation times, AZD5069 appeared to have an antagonist profile with insurmountable antagonism of calcium response curves. This behavior was also observed in vivo in an acute lipopolysaccharide-induced lung inflammation model. Altogether, the data presented here show that AZD5069 represents a novel, potent, and selective CXCR2 antagonist with potential as a therapeutic agent in inflammatory conditions.


Sensitivity analyses of exposure estimates from a quantitative job-exposure matrix (SYN-JEM) for use in community-based studies.
We describe the elaboration and sensitivity analyses of a quantitative job-exposure matrix (SYN-JEM) for respirable crystalline silica (RCS). The aim was to gain insight into the robustness of the SYN-JEM RCS estimates based on critical decisions taken in the elaboration process. SYN-JEM for RCS exposure consists of three axes (job, region, and year) based on estimates derived from a previously developed statistical model. To elaborate SYN-JEM, several decisions were taken: i.e. the application of (i) a single time trend; (ii) region-specific adjustments in RCS exposure; and (iii) a prior job-specific exposure level (by the semi-quantitative DOM-JEM), with an override of 0 mg/m(3) for jobs a priori defined as non-exposed. Furthermore, we assumed that exposure levels reached a ceiling in 1960 and remained constant prior to this date. We applied SYN-JEM to the occupational histories of subjects from a large international pooled community-based case-control study. Cumulative exposure levels derived with SYN-JEM were compared with those from alternative models, described by Pearson correlation ((Rp)) and differences in unit of exposure (mg/m(3)-year). Alternative models concerned changes in application of job- and region-specific estimates and exposure ceiling, and omitting the a priori exposure ranking. Cumulative exposure levels for the study subjects ranged from 0.01 to 60 mg/m(3)-years, with a median of 1.76 mg/m(3)-years. Exposure levels derived from SYN-JEM and alternative models were overall highly correlated (R(p) > 0.90), although somewhat lower when omitting the region estimate ((Rp) = 0.80) or not taking into account the assigned semi-quantitative exposure level (R(p) = 0.65). Modification of the time trend (i.e. exposure ceiling at 1950 or 1970, or assuming a decline before 1960) caused the largest changes in absolute exposure levels (26-33% difference), but without changing the relative ranking ((Rp) = 0.99). Exposure estimates derived from SYN-JEM appeared to be plausible compared with (historical) levels described in the literature. Decisions taken in the development of SYN-JEM did not critically change the cumulative exposure levels. The influence of region-specific estimates needs to be explored in future risk analyses.


Age-associated bidirectional modulation of gene expression in single identified R15 neuron of Aplysia.
Despite the advances in our understanding of aging-associated behavioral decline, relatively little is known about how aging affects neural circuits that regulate specific behaviors, particularly the expression of genes in specific neural circuits during aging. We have addressed this by exploring a peptidergic neuron R15, an identified neuron of the marine snail Aplysia californica. R15 is implicated in reproduction and osmoregulation and responds to neurotransmitters such as acetylcholine, serotonin and glutamate and is characterized by its action potential bursts. We examined changes in gene expression in R15 neurons during aging by microarray analyses of RNAs from two different age groups, mature and old animals. Specifically we find that 1083 ESTs are differentially regulated in mature and old R15 neurons. Bioinformatics analyses of these genes have identified specific biological pathways that are up or downregulated in mature and old neurons. Comparison with human signaling networks using pathway analyses have identified three major networks [(1) cell signaling, cell morphology, and skeletal muscular system development (2) cell death and survival, cellular function maintenance and embryonic development and (3) neurological diseases, developmental and hereditary disorders] altered in old R15 neurons. Furthermore, qPCR analysis of single R15 neurons to quantify expression levels of candidate regulators involved in transcription (CREB1) and translation (S6K) showed that aging is associated with a decrease in expression of these regulators, and similar analysis in three other neurons (L7, L11 and R2) showed that gene expression change during aging could be bidirectional. We find that aging is associated with bidirectional changes in gene expression. Detailed bioinformatics analyses and human homolog searches have identified specific biological processes and human-relevant signaling pathways in R15 that are affected during aging. Evaluation of gene expression changes in different neurons suggests specific transcriptomic signature of single neurons during aging.


Metabolic impact of anti-angiogenic agents on U87 glioma cells.
Glioma cells not only secrete high levels of vascular endothelial growth factor (VEGF) but also express VEGF receptors (VEGFR), supporting the existence of an autocrine loop. The direct impact on glioma cells metabolism of drugs targeting the VEGF pathway, such as Bevacizumab (Bev) or VEGFR Tyrosine Kinase Inhibitor (TKI), is poorly known. U87 cells were treated with Bev or SU1498, a selective VEGFR2 TKI. VEGFR expression was checked with FACS flow cytometry and Quantitative Real-Time PCR. VEGF secretion into the medium was assessed with an ELISA kit. Metabolomic studies on cells were performed using High Resolution Magic Angle Spinning Spectroscopy (HR-MAS). U87 cells secreted VEGF and expressed low level of VEGFR2, but no detectable VEGFR1. Exposure to SU1498, but not Bev, significantly impacted cell proliferation and apoptosis. Metabolomic studies with HR MAS showed that Bev had no significant effect on cell metabolism, while SU1498 induced a marked increase in lipids and a decrease in glycerophosphocholine. Accordingly, accumulation of lipid droplets was seen in the cytoplasm of SU1498-treated U87 cells. Although both drugs target the VEGF pathway, only SU1498 showed a clear impact on cell proliferation, cell morphology and metabolism. Bevacizumab is thus less likely to modify glioma cells phenotype due to a direct therapeutic pressure on the VEGF autocrine loop. In patients treated with VEGFR TKI, monitoring lipids with magnetic resonance spectroscopic (MRS) might be a valuable marker to assess drug cytotoxicity.


Cigarette Smoke or Motor Vehicle Exhaust Exposure Induces PD-L1 Upregulation in Lung Epithelial Cells in COPD Model Rats.
A high smoking-independent chronic obstructive pulmonary disease (COPD) prevalence is observed in lung cancer patients. However, the underlying connection between these two diseases still remains unclear. Cigarette smoking and ambient air pollution are common risk factors for COPD and lung cancer. In this study, we established rat COPD model through exposure to cigarette smoke (CS) or motor vehicle exhaust (MVE). The model rats developed COPD-like phenotypes, manifested as lung functions decline, lung inflammation, emphysema-like alveolar enlargement and airway remodeling. The programmed death-ligand 1 (PD-L1), a factor contributing to immune escape of tumor cells, was overexpressed in lungs from COPD model rats, though more severe COPD phenotypes did not bring with further PD-L1 overexpression in lung. The upregulations of proinflammatory cytokines and PD-L1 were also observed in cultured human bronchial epithelial cells BEAS-2B upon treatment with cigarette smoke extract (CSE) or diesel-related particulate matter 2.5 (PM2.5, SEM1650b). The inflammatory cytokines produced in BEAS-2B cells reflected the PD-L1 levels. Furthermore, ERK1/2, a kinase mediating PD-L1 upregulation in premalignant bronchial cells or NSCLC cells, and STAT1/3, which was reportedly associated with PD-L1 expression in lung tumors, were activated in COPD rats' lungs or in BEAS-2B cells treated with CSE or PM2.5. Therefore, we proposed that inflammation associated PD-L1 overexpression in airway epithelial cells could be the underlying factor facilitating lung cancer incidence in COPD.


Estimation of compressed breast thickness during mammography.
To estimate radiation dose during mammography the breast thickness must be known. We present a new method for estimating the thickness of a compressed breast using only the breast image as projected onto a mammogram, calibration data such as the mAs value and image processing techniques. The method proves to be of high accuracy (+/- 0.2 cm for craniocaudal mammograms) and has the advantage over other methods of allowing retrospective estimation of thickness.


The development of a palliative care program.
Palliative care, which neither hastens death nor prolongs life, provides comfort through the final stages of life. The focus is on comfort for the patient and quality of life for both patient and family. A three-month pilot project used the team approach, involving a physician, nurse, and social worker, to assess how the palliative care needs of the terminal cancer patient were being met at the Cleveland Clinic. Data are presented to demonstrate that such needs exist and are not currently being met by existing services, as perceived by the patient population surveyed. During the pilot project, the palliative care team was in compliance with the Joint Commission on Accreditation of Hospitals hospice standards.


The geographic and demographic scope of shared sanitation: an analysis of national survey data from low- and middle-income countries.
A large and growing proportion of the world's population rely on shared sanitation facilities that have historically been excluded from international targets due to concerns about acceptability, hygiene and access. In connection with a proposed change in such policy, we undertook this study to describe the prevalence and scope of households that report relying on shared sanitation and to characterise them in terms of selected socio-economic and demographic covariates. We extracted data from the most recent national household surveys of 84 low- and middle-income countries from Demographic and Health Surveys and Multiple Indicator Cluster Surveys. We describe the prevalence of shared sanitation and explore associations between specified covariates and reliance on shared sanitation using log-binomial regression. While household reliance on any type of shared sanitation is relatively rare in Europe (2.5%) and the Eastern Mediterranean (7.7%), it is not uncommon in the Americas (14.2%), Western Pacific (16.4%) and South-East Asia (31.3%), and it is most prevalent in Africa (44.6%) where many shared facilities do not meet the definition of 'improved' even if they were not shared (17.7%). Overall, shared sanitation is more common in urban (28.6%) than in rural settings (25.9%), even after adjusting for wealth. While results vary geographically, people who rely on shared sanitation tend to be poorer, reside in urban areas and live in households with more young children and headed by people with no formal education. Data from 21 countries suggest that most sharing is with neighbours and other acquaintances (82.0%) rather than the public. The determinants of shared sanitation identified from these data suggest potential confounders that may explain the apparent increased health risk from sharing and should be considered in any policy recommendation. Both geographic and demographic heterogeneity indicate the need for further research to support a change in policies.


Progesterone inhibition of Xenopus oocyte adenylate cyclase is not mediated via the Bordetella pertussis toxin substrate.
Preincubation of intact Xenopus laevis oocytes with islet-activating protein (IAP) stimulates plasma membrane adenylate cyclase activity measured in the presence of either GTP or 5'-guanylyl imidodiphosphate. The toxin appears to act by catalyzing the ADP-ribosylation of a 41,000-D protein in the oocyte plasma membrane that is distinct from the 45,000- and 52,000-D proteins labeled by cholera toxin. Preincubation of intact oocytes with IAP for 2.5 hr inhibits 80% of the ADP-ribosylation of oocyte membranes subsequently prepared and treated in vitro with IAP in the presence of [alpha-32P]NAD. The abilities of progesterone and the P site agonist, 2',5'-dideoxyadenosine, to inhibit plasma membrane adenylate cyclase are not inhibited by IAP treatment, but IAP pretreatment abolishes inhibition by acetylcholine. In addition, IAP pretreatment of oocytes slows the time course of germinal vesicle breakdown induced by suboptimal concentrations of progesterone. This effect of toxin upon oocyte maturation is apparently due to its effect upon basal cyclase activity, since IAP has no effect upon the time course of maturation induced by 10 microM progesterone. These results indicate that, even though the oocyte adenylate cyclase does contain a functional Ni subunit, inhibition of Xenopus oocyte adenylate cyclase by progesterone or 2',5'-dideoxyadenosine is not mediated via the Bordetella pertussis toxin substrate.


Phylogenomic Reconstruction of the Oomycete Phylogeny Derived from 37 Genomes.
The oomycetes are a class of microscopic, filamentous eukaryotes within the Stramenopiles-Alveolata-Rhizaria (SAR) supergroup which includes ecologically significant animal and plant pathogens, most infamously the causative agent of potato blight Phytophthora infestans. Single-gene and concatenated phylogenetic studies both of individual oomycete genera and of members of the larger class have resulted in conflicting conclusions concerning species phylogenies within the oomycetes, particularly for the large Phytophthora genus. Genome-scale phylogenetic studies have successfully resolved many eukaryotic relationships by using supertree methods, which combine large numbers of potentially disparate trees to determine evolutionary relationships that cannot be inferred from individual phylogenies alone. With a sufficient amount of genomic data now available, we have undertaken the first whole-genome phylogenetic analysis of the oomycetes using data from 37 oomycete species and 6 SAR species. In our analysis, we used established supertree methods to generate phylogenies from 8,355 homologous oomycete and SAR gene families and have complemented those analyses with both phylogenomic network and concatenated supermatrix analyses. Our results show that a genome-scale approach to oomycete phylogeny resolves oomycete classes and individual clades within the problematic Phytophthora genus. Support for the resolution of the inferred relationships between individual Phytophthora clades varies depending on the methodology used. Our analysis represents an important first step in large-scale phylogenomic analysis of the oomycetes. IMPORTANCE The oomycetes are a class of eukaryotes and include ecologically significant animal and plant pathogens. Single-gene and multigene phylogenetic studies of individual oomycete genera and of members of the larger classes have resulted in conflicting conclusions concerning interspecies relationships among these species, particularly for the Phytophthora genus. The onset of next-generation sequencing techniques now means that a wealth of oomycete genomic data is available. For the first time, we have used genome-scale phylogenetic methods to resolve oomycete phylogenetic relationships. We used supertree methods to generate single-gene and multigene species phylogenies. Overall, our supertree analyses utilized phylogenetic data from 8,355 oomycete gene families. We have also complemented our analyses with superalignment phylogenies derived from 131 single-copy ubiquitous gene families. Our results show that a genome-scale approach to oomycete phylogeny resolves oomycete classes and clades. Our analysis represents an important first step in large-scale phylogenomic analysis of the oomycetes.


Transcriptome and metabolite profiling reveals that prolonged drought modulates the phenylpropanoid and terpenoid pathway in white grapes (Vitis vinifera L.).
Secondary metabolism contributes to the adaptation of a plant to its environment. In wine grapes, fruit secondary metabolism largely determines wine quality. Climate change is predicted to exacerbate drought events in several viticultural areas, potentially affecting the wine quality. In red grapes, water deficit modulates flavonoid accumulation, leading to major quantitative and compositional changes in the profile of the anthocyanin pigments; in white grapes, the effect of water deficit on secondary metabolism is still largely unknown. In this study we investigated the impact of water deficit on the secondary metabolism of white grapes using a large scale metabolite and transcript profiling approach in a season characterized by prolonged drought. Irrigated grapevines were compared to non-irrigated grapevines that suffered from water deficit from early stages of berry development to harvest. A large effect of water deficit on fruit secondary metabolism was observed. Increased concentrations of phenylpropanoids, monoterpenes, and tocopherols were detected, while carotenoid and flavonoid accumulations were differentially modulated by water deficit according to the berry developmental stage. The RNA-sequencing analysis carried out on berries collected at three developmental stages-before, at the onset, and at late ripening-indicated that water deficit affected the expression of 4,889 genes. The Gene Ontology category secondary metabolic process was overrepresented within up-regulated genes at all the stages of fruit development considered, and within down-regulated genes before ripening. Eighteen phenylpropanoid, 16 flavonoid, 9 carotenoid, and 16 terpenoid structural genes were modulated by water deficit, indicating the transcriptional regulation of these metabolic pathways in fruit exposed to water deficit. An integrated network and promoter analyses identified a transcriptional regulatory module that encompasses terpenoid genes, transcription factors, and enriched drought-responsive elements in the promoter regions of those genes as part of the grapes response to drought. Our study reveals that grapevine berries respond to drought by modulating several secondary metabolic pathways, and particularly, by stimulating the production of phenylpropanoids, the carotenoid zeaxanthin, and of volatile organic compounds such as monoterpenes, with potential effects on grape and wine antioxidant potential, composition, and sensory features.


The role of weather in mediating the effect of mercury exposure on reproductive success in tree swallows.
Mercury is a heavy metal that has contaminated countless ecosystems throughout the world. A large body of literature has documented reproductive, physiological, and behavioral impairments associated with mercury exposure in laboratory settings, but whether and how such effects are manifest in free-living populations remains poorly understood. The purpose of this study was to evaluate whether tree swallow (Tachycineta bicolor) breeding success at a site with high mercury exposure varied with ambient temperature or precipitation at various points in the breeding cycle. Tree swallows nesting along the South River had significantly elevated blood total mercury (mean ± SE: 3.03 ± 0.15 μg/g) compared to swallows breeding on reference sites (mean ± SE: 0.16 ± 0.005 μg/g). These high levels of mercury were associated with reduced hatching and fledging success, and contaminated birds produced approximately one less fledgling per nest than their reference counterparts. The magnitude of this difference was weather-dependent: unusually high ambient temperatures encountered early in the nestling period were associated with reduced reproductive output in contaminated, but not in reference, birds. In contrast, little effect of mercury on success of nestlings was observed when temperatures were cooler, and precipitation also had no detectable interaction with mercury. These results provide insight into mechanisms underlying reproductive effects of mercury. In addition, these findings underscore the importance of considering variable environmental conditions when assessing effects of contaminants on free-living wildlife. In particular, projections about the effects of global climate change on ecotoxicological impacts must take into account the kinds of weather-mediated effect demonstrated here.


Different mechanisms define the antiadhesive function of high molecular weight kininogen in integrin- and urokinase receptor-dependent interactions.
Proteolytic cleavage of single-chain high molecular weight kininogen (HK) by kallikrein releases the short-lived vasodilator bradykinin and leaves behind 2-chain high molecular weight kininogen (HKa) that has been previously reported to exert antiadhesive properties as well as to bind to the urokinase receptor (uPAR) on endothelial cells. In this study we defined the molecular mechanisms for the antiadhesive effects of HKa related to disruption of integrin- and uPAR-mediated cellular interactions. Vitronectin (VN) but not fibrinogen or fibronectin-dependent alphavbeta(3) integrin-mediated adhesion of endothelial cells was blocked by HKa or its isolated domain 5. In a purified system, HKa but not HK competed for the interaction of VN with alphavbeta(3) integrin, because HKa and the isolated domain 5 but not HK bound to both multimeric and native VN in a Zn(2+)-dependent manner. The interaction between HKa or domain 5 with VN was prevented by heparin, plasminogen activator inhibitor-1, and a recombinant glutathione-S-transferase (GST)-fusion peptide GST-VN (1-77) consisting of the amino terminal portion of VN (amino acids 1-77), but not by a cyclic arginyl-glycyl-aspartyl peptide, indicating that HKa interacts with the amino terminal portion of VN ("somatomedin B region"). Furthermore, we have confirmed that HKa but not HK bound to uPAR and to the truncated 2-domain form of uPAR lacking domain 1 in a Zn(2+)-dependent manner. Through these interactions, HKa or its recombinant His-Gly-Lys-rich domain 5 completely inhibited the uPAR-dependent adhesion of myelomonocytic U937 cells and uPAR-transfected BAF-3 cells to VN and thereby promoted cell detachment. By immunogold electron microscopy, both VN and HK/HKa were found to be colocalized in sections from human atherosclerotic coronary artery, indicating that the described interactions are likely to take place in vivo. Taken together, HK and HKa inhibit different VN-responsive adhesion receptor systems and may thereby influence endothelial cell- or leukocyte-related interactions in the vasculature, particularly under inflammatory conditions. (Blood. 2000;96:514-522)


Adhesion of polyelectrolyte microcapsules through biotin-streptavidin specific interaction.
Adhesion of PAH/PSS and PDADMAC/PSS capsules through electrostatic and specific interactions has been investigated using reflective interference contrast microscopy (RICM). Adhesion of capsules via electrostatic interactions was found to be spontaneous and strong. Capsules functionalized with poly(l-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) did not exhibit significant adhesion (as determined by the adhesion area) to streptavidin-coated substrates, whereas capsules functionalized with biotinylated PLL-g-PEG showed a significantly larger adhesion area. Using continuum mechanical models, the total adhesion energies for these cases were calculated and were found to correspond to several tens of individual biotin-streptavidin pairs. The application of specific interactions such as the biotin-streptavidin system for controlled capsule adhesion has been demonstrated in this study.


Microcalcifications in breast cancer: an active phenomenon mediated by epithelial cells with mesenchymal characteristics.
Mammary microcalcifications have a crucial role in breast cancer detection, but the processes that induce their formation are unknown. Moreover, recent studies have described the occurrence of the epithelial-mesenchymal transition (EMT) in breast cancer, but its role is not defined. In this study, we hypothesized that epithelial cells acquire mesenchymal characteristics and become capable of producing breast microcalcifications. Breast sample biopsies with microcalcifications underwent energy dispersive X-ray microanalysis to better define the elemental composition of the microcalcifications. Breast sample biopsies without microcalcifications were used as controls. The ultrastructural phenotype of breast cells near to calcium deposits was also investigated to verify EMT in relation to breast microcalcifications. The mesenchymal phenotype and tissue mineralization were studied by immunostaining for vimentin, BMP-2, β2-microglobulin, β-catenin and osteopontin (OPN). The complex formation of calcium hydroxyapatite was strictly associated with malignant lesions whereas calcium-oxalate is mainly reported in benign lesions. Notably, for the first time, we observed the presence of magnesium-substituted hydroxyapatite, which was frequently noted in breast cancer but never found in benign lesions. Morphological studies demonstrated that epithelial cells with mesenchymal characteristics were significantly increased in infiltrating carcinomas with microcalcifications and in cells with ultrastructural features typical of osteoblasts close to microcalcifications. These data were strengthened by the rate of cells expressing molecules typically involved during physiological mineralization (i.e. BMP-2, OPN) that discriminated infiltrating carcinomas with microcalcifications from those without microcalcifications. We found significant differences in the elemental composition of calcifications between benign and malignant lesions. Observations of cell phenotype led us to hypothesize that under specific stimuli, mammary cells, which despite retaining a minimal epithelial phenotype (confirmed by cytokeratin expression), may acquire some mesenchymal characteristics transforming themselves into cells with an osteoblast-like phenotype, and are able to contribute to the production of breast microcalcifications.


Monocyte-mediated serum-independent damage to hyphal and pseudohyphal forms of Candida albicans in vitro.
Human peripheral blood monocytes attached to Candida albicans hyphae in the absence of serum and damaged the hyphae without completely ingesting them. Attachment and damage was not augmented by the addition of serum. Damage to hyphae was quantitated by a previously developed metabolic assay that measured leukocyte-induced reduction in uptake of [(14)C]cytosine by the hyphae. Use of cells from patients with hereditary disorders of leukocyte function, chronic granulomatous disease, and myeloperoxidase deficiency indicated that myeloperoxidase-independent and nonoxidative mechanisms could sometimes damage hyphae where oxidative mechanisms were impaired. Damage to hyphae by normal monocytes was inhibited by concentrations of sodium azide and sodium cyanide that primarily affect myeloperoxidase activity, as well as by halide-free conditions, catalase, and putative antagonists of hypochlorous acid or singlet oxygen. Iodination of hyphae, a myeloperoxidase and hydrogen peroxide-dependent process of monocytes, was similarly inhibited by sodium azide, sodium cyanide, and catalase. Under anaerobic conditions, damage to hyphae was reduced by 64.0-68.4%. In contrast, inhibitors of potential nonoxidative antifungal mechanisms, iron salts to saturate iron chelators, and polyanionic amino acid polymers to neutralize cationic proteins did not block damage to hyphae by monocytes. Preparations rich in lysosomal granules from fractionated normal monocytes also did not damage hyphae. Overall, it appeared that oxidative mechanisms were most important for damage to hyphae by normal monocytes. Electron microscopy confirmed that Candida hyphae were damaged and probably killed by monocytes, but monocytes appeared to sustain significant damage in the process. In the absence of serum, monocyte cell membranes became closely approximated to Candida cell walls. It appeared that some Candida could escape this partial engulfment, as they were seen floating free with vesicular trilaminar membrane remnants covering hyphal surfaces. In general, monocytes appeared to be damaged by interactions with Candida hyphae more than neutrophils had been in previous studies.


NKG2D Ligand Shedding in Response to Stress: Role of ADAM10.
NKG2D is an activating receptor expressed by NK cells and some subsets of T cells and represents a major recognition receptor for detection and elimination of cancer cells. The ligands of NKG2D are stress-induced self-proteins that can be secreted as soluble molecules by protease-mediated cleavage. The release of NKG2D ligands in the extracellular milieu is considered a mode of finely controlling their surface expression levels and represents a relevant immune evasion mechanism employed by cancer cells to elude NKG2D-mediated immune surveillance. A disintegrin and metalloproteinase 10 (ADAM10), a catalytically active member of the ADAM family of proteases, is involved in the cleavage of some NKG2D ligands in various types of cancer cells either in steady state conditions and in response to an ample variety of stress stimuli. Appealing immunotherapeutic strategies devoted to promoting NK cell-mediated recognition and elimination of cancer cells are based on the upregulation of NK cell activating ligands. In particular, activation of DNA damage response (DDR) and the induction of cellular senescence by chemotherapeutic agents are associated with increased expression of NKG2D ligands on cancer cell surface. Herein, we will review advances on the protease-mediated cleavage of NKG2D ligands in response to chemotherapy-induced stress focusing on: (i) the role played by ADAM10 in this process and (ii) the implications of NKG2D ligand shedding in the course of cancer therapy and in senescent cells.


Dynamics of nuclear lamina assembly and disassembly.
What can be concluded about lamin dynamics? a. While the nuclear lamina forms a tight network of proteins, individual lamina members, such as the lamin C proteins, are only partially bound to the lamina. b. A prominent pool of nucleoplasmic lamins exists in most cells, which interacts with intranuclear structures (DNA? Histones? Replication and/or transcription complexes?) in a dynamic fashion. c. During mitosis lamins do not play a key role in the initial reformation of the nuclear envelope. However, they are important for the correct functioning of the nucleus immediately after mitosis. Many questions remain unanswered, although nuclear lamins have been studied at different levels for over 20 years. We have only just begun to understand their crucial role in several cellular processes. Some of the important questions that still remain are: a. The function of intranuclear lamin foci. Are these native nuclear structures (nuclear channels/tubules) and do they play a role in replication and transcription? b. The function of the nucleoplasmic veil of lamins. Are these molecules only temporarily bound to intranuclear structures as a result of affinity to chromosomes or nuclear proteins, or is there a functional interaction with these molecules? c. The extent to which lamin molecules add to nuclear membrane organisation. Do these molecules play a key role in keeping the membrane intact and the nucleus functional, or are they only one of many supportive component of the nuclear membrane?


Evidence for specific secretion rather than autolysis in the release of some Helicobacter pylori proteins.
We investigated whether Helicobacter pylori cells actively secrete proteins such as the urease subunits UreA and UreB and the GroES and GroEL homologs HspA and HspB or whether these proteins were present in the extracellular compartment as a consequence of autolysis. Using a subcellular fractionation approach associated with quantitative Western blot analyses, we showed that the supernatant protein profiles were very different from those of the cell pellets, even for bacteria harvested in the late growth phase; this suggests that the release process is selective. A typical cytoplasmic protein, a beta-galactosidase homolog, was found exclusively associated with the pellet of whole-cell extracts, and no traces were found in the supernatant. In contrast, UreA, UreB, HspA, and HspB were mostly found in the pellet but significant amounts were also present in the supernatant. HspA and UreB were released into the supernatant at the same rate throughout the growth phase (3%), whereas large portions of HspB and UreA were released during the stationary phase (over 30 and 20%, respectively) rather than during the early growth phase (20% and 6, respectively). The profiles of protein obtained after water extraction of the bacteria with those of the proteins naturally released within the liquid culture supernatants demonstrated that water extraction led to the release of a large amount of protein due to artifactual lysis. Our data support the conclusion that a specific and selective mechanism(s) is involved in the secretion of some H. pylori antigens. A programmed autolysis process does not seem to make a major contribution.


Hepatocellular Carcinoma Patients May Benefit From Postoperative Huaier Aqueous Extract After Liver Transplantation.
Liver transplantation has been the first choice for most early- or intermediate-stage hepatocellular carcinoma (HCC) cases. However, postoperative anti-HCC therapies remain controversial. In this study, we aimed to evaluate the safety and efficacy of Huaier aqueous extract (Jinke), when used as an adjuvant postoperative anti-HCC therapy. We retrospectively collected the clinical and follow-up data of HCC patients who underwent liver transplantation at our center. We divided them into 2 groups: a control liver transplantation group and a Huaier treatment group. The baseline characteristics, tumor characteristics, intraoperative data, postoperative recovery, long-term overall survival rate, and tumor-free survival rate were compared between the 2 groups. Fifty-three patients were included in our study, including 28 patients who underwent postoperative Huaier therapy and 25 patients who underwent liver transplantation without postoperative Huaier therapy. The baseline and tumor characteristics were similar between the 2 groups. None of the patients in the Huaier group experienced any severe adverse events. The long-term predictive overall survival was similar between the 2 groups (P = .202). However, the Huaier group had a higher predictive tumor-free survival rate than the control group (P = .029). And the 10- and 30-month predictive tumor recurrence rates were 17.9% and 35.7% in the Huaier group, which were significantly lower than those in the control group (60% and 64%; P < .05). HCC patients may benefit from Huaier therapy after liver transplantation, but a longer follow-up time and larger cohort study may be necessary to be sure.


Apolipoprotein E peptide stimulation of rat ovarian theca cell androgen synthesis is mediated by members of the low density lipoprotein receptor superfamily.
Ovarian androgen production is rate limiting for follicular maturation and can induce follicular atresia. Thus, it is important to define the actions of the intraovarian agents, such as apolipoprotein (apo) E, that modulate theca cell androgen production. Theca cell androgen production is stimulated at low concentrations and inhibited at higher concentrations of native apo E. The apo E peptide, acetyl-Y(LRKLRKRLLRDADDL)(2)C or acetyl-Y(141-155)(2)C, has low density lipoprotein (LDL) receptor and LDL receptor-related protein-binding activity, and it mimics the activity of native apo E in the theca-interstitial cell system. To define the role of members of the LDL receptor superfamily in the apo E peptide-mediated responses, we found that receptor-associated protein prevented the stimulation without altering the inhibition of androstenedione production. The apo E peptide (129-162), which has no LDL receptor-binding activity, did not stimulate androstenedione production. The apo E peptide acetyl-Y(141-155)(2)C did not stimulate androstenedione production when cell surface heparan sulfate proteoglycans were degraded with heparinase. The apo E peptide acetyl-Y(141-155)(2)C bound to heparin, a property of LDL receptor ligands, and in this complex the peptide had no effect on androstenedione production. These observations support the conclusion that apo E-mediated stimulation, but not inhibition, of ovarian theca cell androstenedione production was mediated by members of the LDL receptor superfamily.


Effective Model Update for Adaptive Classification of Text Streams in a Distributed Learning Environment.
In this study, we propose dynamic model update methods for the adaptive classification model of text streams in a distributed learning environment. In particular, we present two model update strategies: (1) the entire model update and (2) the partial model update. The former aims to maximize the model accuracy by periodically rebuilding the model based on the accumulated datasets including recent datasets. Its learning time incrementally increases as the datasets increase, but we alleviate the learning overhead by the distributed learning of the model. The latter fine-tunes the model only with a limited number of recent datasets, noting that the data streams are dependent on a recent event. Therefore, it accelerates the learning speed while maintaining a certain level of accuracy. To verify the proposed update strategies, we extensively apply them to not only fully trainable language models based on CNN, RNN, and Bi-LSTM, but also a pre-trained embedding model based on BERT. Through extensive experiments using two real tweet streaming datasets, we show that the entire model update improves the classification accuracy of the pre-trained offline model; the partial model update also improves it, which shows comparable accuracy with the entire model update, while significantly increasing the learning speed. We also validate the scalability of the proposed distributed learning architecture by showing that the model learning and inference time decrease as the number of worker nodes increases.


Endocannabinoid system in food intake and metabolic regulation.
As the incidence of obesity and the metabolic syndrome has increased, research has focused on the importance of the endocannabinoid system in the brain and peripheral tissues. Rimonabant, an inverse agonist of the CB1 receptor is being used therapeutically. This review presents recent advances in endocannabinoid physiology. The endocannabinoid system interacts with other anorexigenic and orexigenic pathways to regulate food intake in the hypothalamus, and the hedonistic value of food in the mesolimbic system. Endocannabinoid system overactivity contributes to hepatic steatosis, increased adipose tissue inflammation, dysregulated insulin signalling in the pancreas and disturbed oxidative pathways in skeletal muscle. The breakdown pathways for anandamide and 2-arachidonoylglycerol, the endocannabinoid receptor ligands, are reviewed, and the recent discoveries of endocannabinoid receptor polymorphisms and their relationship to obesity and metabolic disease noted. The favourable effect of rimonabant on fat mass glycaemic control, lipid metabolism and overall cardiovascular risk must be tempered by adverse effects on mood. The ubiquitous role of the endocannabinoid system in food intake and energy metabolism is now established. Drugs that manipulate different aspects of this system may benefit subjects with the metabolic and cachectic syndromes.


Exploring the impact of patient-specific clinical features on osimertinib effectiveness in a real-world cohort of patients with EGFR mutated non-small cell lung cancer.
Osimertinib is prescribed to patients with metastatic non-small cell lung cancer (NSCLC) and a sensitizing EGFR mutation. Limited data exists on the impact of patient characteristics or osimertinib exposure on effectiveness outcomes. This was a Dutch, multicenter cohort study. Eligible patients were ≥18 years, with metastatic EGFRm+ NSCLC, receiving osimertinib. Primary endpoint was progression-free survival (PFS). Secondary endpoints included overall survival (OS) and safety. Kaplan-Meier analyses and multivariate Cox proportional hazard models were performed. In total, 294 patients were included. Primary EGFR-mutations were mainly exon 19 deletions (54%) and p.L858R point mutations (30%). Osimertinib was given in first-line (40%), second-line (46%) or beyond (14%), with median PFS 14.4 (95% CI: 9.4-19.3), 13.9 (95% CI: 11.3-16.1) and 8.7 months (95% CI: 4.6-12.7), respectively. Patients with low BMI (<20.0 kg/m2 ) had significantly shorter PFS/OS compared to all other subgroups. Patients with a high plasma trough concentration in steady state (Cmin,SS ; >271 ng/mL) had shorter PFS compared to a low Cmin,SS (<163 ng/mL; aHR 2.29; 95% CI: 1.13-4.63). A significant longer PFS was seen in females (aHR = 0.61, 95% CI: 0.45-0.82) and patients with the exon 19 deletion (aHR = 0.58, 95% CI: 0.36-0.92). A trend towards longer PFS was seen for TP53 wild-type patients, while age did not impact PFS. Patients with a primary EGFR exon 19 deletion had longer PFS, while a low BMI, male sex and a high Cmin,SS were indicative for shorter PFS and/or OS. Age was not associated with effectiveness outcomes of osimertinib.


Esophageal function, radiography, and dysphagia in Sjögren's syndrome.
Esophageal function and anatomy were investigated with manometry, acid perfusion test, acid clearing test, and x-ray in 11 patients with primary Sjögren's syndrome (SS) and in 11 with secondary SS. The manometric investigation revealed minor motor differences in the SS patients as compared to 16 controls, ie, shorter peristaltic contraction time of the whole esophagus, and faster peristaltic velocity preferably in the distal part of the esophagus, while the results from the reflux tests did not differ between patients and controls. Radiographic examination revealed upper esophageal webs in 10% (2/20), and hiatal hernia in 25% (5/20). The dysphagia as reported by 73% of the patients cannot be explained by webs or impaired motor function and is regarded to be secondary to lack of saliva, making the solid bolus passage difficult.


Preparation and evaluation of silica-based ionic liquid-modified stationary phase for HPLC.
A new high-performance liquid chromatography (HPLC) stationary phase was firstly synthesized based on the ionic liquid 2-ethyl-4-methylimidazole chloride. And the performances of the resulting stationary phase were determined by Fourier transform infrared (FT-IR), thermogravimetric analysis (TGA), and (13)C nuclear magnetic resonance (NMR) spectroscopy. The retention characteristics of the resulting stationary phase were evaluated by normal phase HPLC conditions to separate tanshinone I and tanshinone IIA in Danshen herb. The results demonstrated that the synthesized ionic liquid was bond-linked to the silica surface successfully and that IL-modified silica is more effective than unmodified silica when separating tanshinone I and tanshinone IIA in Danshen herb.


Long-term consequences of obesity on female fertility and the health of the offspring.
Obesity has reached near epidemic levels among reproductive age women with a myriad of consequences. Obesity adversely affects the maternal milieu by creating conditions that decrease fertility and increase the risk of gestational diabetes, hypertensive disease in pregnancy, fetal growth abnormalities and congenital anomalies. The effects of obesity are not limited to pregnancy. Indeed, beyond the immediate postpartum period, obese women maintain a higher prevalence of insulin resistance and cardiovascular disease. In this article, we will review the pathophysiology underlying the effects of obesity on fertility, pregnancy outcome and health status of offspring. The purpose of this review is to outline proposed models responsible for the short-term and long-term consequences of obesity on fertility and offspring development, and identify knowledge gaps where additional research is needed. Maternal over or under nutrition adversely affect maternal reproductive capacity and pregnancy success. Separate from effects on maternal reproductive function, maternal over or under nutrition may also 'program' fetal pathophysiology through inheritance mechanisms that suggest epigenetic modification of DNA, differential RNA translation and protein expression, or modification of the fetal hypothalamic-pituitary axis function through programmed adverse effects on the developing hypothalamic circuitry. The concept of maternal health modifying the risk of developing noncommunicable diseases in the offspring is based on Developmental Origins of Health and Disease hypothesis. Of importance, the long-term effects of obesity are not limited to maternal health, but also programs pathophysiology in their offspring. Children of obese gravida are at increased risk for the development of cardiometabolic disease in childhood and throughout adulthood. Future studies directly interrogating mechanisms underlying the risks associated with obesity will allow us to develop interventions and therapies to decrease short-term and long-term morbidities associated with maternal obesity.


PTEN is essential for cell migration but not for fate determination and tumourigenesis in the cerebellum.
PTEN is a tumour suppressor gene involved in cell cycle control, apoptosis and mediation of adhesion and migration signalling. Germline mutations of PTEN in humans are associated with familial tumour syndromes, among them Cowden disease. Glioblastomas, highly malignant glial tumours of the central nervous system frequently show loss of PTEN. Recent reports have outlined some aspects of PTEN function in central nervous system development. Using a conditional gene disruption approach, we inactivated Pten in mice early during embryogenesis locally in a region specific fashion and later during postnatal development in a cell-specific manner, to study the role of PTEN in differentiation, migration and neoplastic transformation. We show that PTEN is required for the realisation of normal cerebellar architecture, for regulation of cell and organ size, and for proper neuronal and glial migration. However, PTEN is not required for cell differentiation and lack of PTEN is not sufficient to induce neoplastic transformation of neuronal or glial cells


[Sheltering: the perception of women on the trajectory of childbearing].
The goal of the study was to know the expectation and perception of women about hospital care during childbirth. It is a qualitative research, of the descriptive-exploratory kind; it was developed on a teaching hospital, having as participants fourteen puerperal women. To the data collection a semi-structured interviews was used; to analyze and interpretation the information we used the Method of Qualitative Analysis of a Situated Phenomenon. The results indicated as one of the themes Perceiving the need of sheltering and welcoming on the trajectory of childbearing which is presented on this article. The obtained information reveal that women expect hospitalar sheltering and welcoming at the institutional, interpersonal and technical dimensions.


Time-resolved luminescent biosensing based on inorganic lanthanide-doped nanoprobes.
Time-resolved (TR) photoluminescence (PL) biosensing has been widely adopted in many research and medical institutions. However, commercial molecular TRPL bioprobes like lanthanide (Ln(3+))-chelates suffer from poor photochemical stability and long-term toxicity. Inorganic Ln(3+)-doped nanocrystals (NCs), owing to their superior physicochemical properties over Ln(3+)-chelates, are regarded as a new generation of luminescent nanoprobes for TRPL biosensing. The long-lived PL of Ln(3+)-doped NCs combined with the TRPL technique is able to completely suppress the interference of the short-lived background, resulting in a background-free signal and therefore a remarkable sensitivity for biosensing. In this feature article, we summarize the latest advancements in inorganic Ln(3+)-doped NCs as TRPL nano-bioprobes from their fundamental optical properties to their potential applications for ultrasensitive biodetection and high-resolution bioimaging. Future efforts towards the commercialization of these nanoprobes are also proposed.


Nature, nurture, and ethnocentrism in the Minnesota twin study.
The preponderance of research on the study of ethnocentrism has primarily attributed such attitudes to learned behavior. The research here advances the argument that both socialization and genetic inheritance contribute to the development of ethnocentric attitudes and behavior. This analysis employs the Minnesota Twins Political Survey data consisting of 596 complete twin pairs. Using the classical twin design, we employed structural equation modeling to model the covariance of twins in regards to additive genetic effects, shared environmental effects, and unique environmental effects (i.e., the classic ACE model). The findings reveal that genetic inheritance is significant in explaining the variance in genetic attitudes. Specifically, genetic inheritance explains 18% of the variance, with the overwhelming 82% being explained by the unique environment.


Influence of crystal shape on the tableting performance of L-lysine monohydrochloride dihydrate.
The purpose of this study is to understand the influence of crystal shape on the tableting performance of L-lysine monohydrochloride (LMH) dihydrate, using the method of data analysis developed by Joiris E et al. 1998. Pharm Res 15:1122-1130. Phase-pure crystals of LMH dihydrate, prism-shaped (S) and plate-shaped (T), were prepared by adjusting the composition of the crystallization solvent. At the same compaction pressure, T always gives stronger tablets than S, (i.e.; the tabletability of T is greater). The porosity of tablets from T crystals is always greater than that of S crystals when compressed at the same pressure, (i.e.; the compressibility of T is lower). The tensile strength of T tablets, at the same porosity, is greater than that of S tablets, (i.e.; the compactibility of T is greater). Therefore, the greater tabletability of T is a result of its better compactibility that overcomes the negative effects by its lower compressibility. The greater compactibility of T is related to favorable orientation of the slip planes in the tablet, corresponding to greater plasticity under load. The yield strengths of T and S crystals are essentially the same (20 MPa). Therefore, the crystal shape influences the tableting performance but does not, in principle, affect the yield strength of LMH dihydrate.


Neuromuscular versus quadriceps strengthening exercise in patients with medial knee osteoarthritis and varus malalignment: a randomized controlled trial.
To compare the effects of neuromuscular exercise (NEXA) and quadriceps strengthening (QS) on the knee adduction moment (an indicator of mediolateral distribution of knee load), pain, and physical function in patients with medial knee joint osteoarthritis (OA) and varus malalignment. One hundred patients with medial knee pain, mostly moderate-to-severe radiographic medial knee OA, and varus malalignment were randomly allocated to one of two 12-week exercise programs. Each program involved 14 individually supervised exercise sessions with a physiotherapist plus a home exercise component. Primary outcomes were peak external knee adduction moment (3-dimensional gait analysis), pain (visual analog scale), and self-reported physical function (Western Ontario and McMaster Universities Osteoarthritis Index). Eighty-two patients (38 [76%] of 50 in the NEXA group and 44 [88%] of 50 in the QS group) completed the trial. There was no significant between-group difference in the change in the peak knee adduction moment (mean difference 0.13 Nm/[body weight × height]% [95% confidence interval (95% CI) -0.08, 0.33]), pain (mean difference 2.4 mm [95% CI -6.0, 10.8]), or physical function (mean difference -0.8 units [95% CI -4.0, 2.4]). Neither group showed a change in knee moments following exercise, whereas both groups showed similar significant reductions in pain and improvement in physical function. Although comparable improvements in clinical outcomes were observed with both neuromuscular and quadriceps strengthening exercise in patients with moderate varus malalignment and mostly moderate-to-severe medial knee OA, these forms of exercise did not affect the knee adduction moment, a key predictor of structural disease progression.


COVID-19 Mortality Rates Among Nursing Home Residents Declined From March To November 2020.
Improved therapeutics and supportive care in hospitals have helped reduce mortality from COVID-19. However, there is limited evidence as to whether nursing home residents, who account for a disproportionate share of COVID-19 deaths and are often managed conservatively in the nursing home instead of being admitted to the hospital, have experienced similar mortality reductions. In this study we examined changes in thirty-day mortality rates between March and November 2020 among 12,271 nursing home residents with COVID-19. We found that adjusted mortality rates significantly declined from a high of 20.9 percent in early April to 11.2 percent in early November. Mortality risk declined for residents with both symptomatic and asymptomatic infections and for residents with both high and low clinical complexity. The mechanisms driving these trends are not entirely understood, but they may include improved clinical management within nursing homes, improved personal protective equipment supply and use, and genetic changes in the virus.


The effect of the addition of colloidal iridium oxide into sol-gel obtained titanium and ruthenium oxide coatings on titanium on their electrochemical properties.
Electrochemical properties of sol-gel processed Ti(0.6)Ir(0.4)O(2) and Ti(0.6)Ru(0.3)Ir(0.1)O(2) coatings on titanium substrate were investigated using cyclic voltammetry, polarization measurements and electrochemical impedance spectroscopy and compared to the properties of Ti(0.6)Ru(0.4)O(2) coating. The role of iridium oxide in the improvement of the electrocatalytic, capacitive and stability properties of titanium anodes activated by a RuO(2)-TiO(2) coating is discussed. The oxide sols were prepared by forced hydrolysis of the metal chlorides. The characterization by dynamic light scattering and X-ray diffraction showed that polydisperse oxide sols were obtained with the particles tending to form agglomerates. The presence of IrO(2) causes a suppression of the X-ray diffraction peaks of TiO(2) and RuO(2) in the sol-gel prepared Ti(0.6)Ir(0.4)O(2) and Ti(0.6)Ru(0.3)Ir(0.1)O(2) coatings. The IrO(2)-containing coatings had an enhanced charge storage ability and activity for the oxygen evolution reaction (OER) in comparison to Ti(0.6)Ru(0.4)O(2) coating. The voltammogram of the Ti(0.6)Ir(0.4)O(2)/Ti electrode showed well-resolved peaks related to Ir redox transitions, which are responsible for the enhanced charge storage ability of IrO(2)-containing coatings. Redox transitions of Ir were also registered in the high-frequency domain of the ac impedance spectra of the coatings as a semicircle with characteristics insensitive to the electrolyte composition and to the electrode potential prior to OER. However, the semicircle characteristics were different for the two IrO(2)-containing coatings, as well as at potentials outside the OER in comparison to those at which the OER occurs.


Changes in attitudes to vaccination as a result of the COVID-19 pandemic: A longitudinal study of older adults in the UK.
The rapid development of COVID-19 vaccines has brought an unprecedented focus on public attitudes to vaccines, with intention to accept a COVID-19 vaccine fluctuating during the pandemic. However, it is unclear how the pandemic may influence attitudes and behaviour in relation to vaccines in general. The aim of the current study is to examine older adults' changes in vaccination attitudes and behaviour over the first year of the pandemic. In February-March 2020 (before the first COVID-19 national lockdown in the UK), 372 older adults (aged 65+) provided sociodemographic information, self-reported influenza vaccine uptake, and completed two measures of vaccination attitudes: the 5C scale and the Vaccination Attitudes Examination Scale. One-year later, following rollout of COVID-19 vaccines to older adults, participants provided information on their COVID-19 and influenza vaccine uptake in the previous 12 months, and completed the 5C and VAX scales again. Paired samples t-tests were used to examine changes in vaccination attitudes over time. Almost all participants (98.7%) had received at least one dose of a COVID-19 vaccine, and a significant increase in influenza uptake was identified (83.6% in 2020 to 91.6% in 2021). Complacency, mistrust of vaccine benefit, concerns about commercial profiteering, and constraints to vaccination had significantly decreased between Time 1 and Time 2, and collective responsibility had significant increased. However, calculation and worries about unforeseen future effects had increased, indicating that participants now perceived higher risks related to vaccination and were taking a more deliberative information-seeking approach. The results show significant changes in vaccination attitudes across the pandemic. These changes suggest that while older adults became less complacent about the importance of vaccines, concerns about potential risks associated with vaccination increased. It will be important for public health communication to address these concerns for all vaccines offered to this group.


Coupling of oxytocin receptor to G proteins in rat myometrium during labor: Gi receptor interaction.
Occupancy of oxytocin receptor (OTR) binding sites in pregnant rat myometrial membranes with iodinated oxytocin antagonist (OTA), followed by detergent solubilization and size selection, showed that radioactivity eluted in two distinct peaks: one corresponding in size to the isolated receptor (approximately 60 kDa) and the other ranging from 240 to 320 kDa. The unliganded 240- to 320-kDa fraction contained OTRs coupled to G proteins, as the addition of oxytocin (OT) increased guanosine 35S-labeled 5'-O-(3-thiotriphosphate) binding up to twofold in a dose-dependent manner. The effects of OT were blocked by coincubation with OTA. G protein alpha-subunits associated with OTRs in the 240- to 320-kDa peak were identified by immunoadsorption. Significant amounts of both G alpha q/11 and G alpha i3 were associated with the OTR; a lesser amount of G alpha s was complexed. Using the same approach but with antibodies to effector enzymes, we observed that phospholipase C beta 1 (PLC beta 1) and PLA2 were also associated with the OTR. The results corroborate the well-established interaction of OTR with Gq and further show that Gi coupling might be an important component of OTR signal transduction. To further investigate the interaction of Gi with the OTR, we showed that OT stimulation of guanosine 5'-triphosphatase activity in intact myometrial membranes was inhibited by pertussis toxin. Pertussis toxin-stimulated ADP ribosylation of G alpha i in myometrial membranes was also decreased by OT treatment. These findings with pertussis toxin strongly indicate that OTR is coupled to Gi in rat myometrial membranes. The 60-kDa OTR peak (noncoupled receptor) was demonstrable in the myometrium only before the end of gestation and after parturition and accounted for about one-half the 125I-OTA binding activity. At term, there was about a fivefold increase in binding and almost a complete shift to the 240- to 320-kDa-size complex. Thus the established increased sensitivity of the myometrium to OT at term could be the result of both upregulation of OTRs and an increase in the fraction of receptors coupled to signal transduction components, one of which is Gi.


Cortical theta wanes for language.
The role of low frequency oscillations in language areas is not yet understood. Using ECoG in six human subjects, we studied whether different language regions show prominent power changes in a specific rhythm, in similar manner as the alpha rhythm shows the most prominent power changes in visual areas. Broca's area and temporal language areas were localized in individual subjects using fMRI. In these areas, the theta rhythm showed the most pronounced power changes and theta power decreased significantly during verb generation. To better understand the role of this language-related theta decrease, we then studied the interaction between low frequencies and local neuronal activity reflected in high frequencies. Amplitude-amplitude correlations showed that theta power correlated negatively with high frequency activity, specifically across verb generation trials. Phase-amplitude coupling showed that during control trials, high frequency power was coupled to theta phase, but this coupling decreased significantly during verb generation trials. These results suggest a dynamic interaction between the neuronal mechanisms underlying the theta rhythm and local neuronal activity in language areas. As visual areas show a pronounced alpha rhythm that may reflect pulsed inhibition, language regions show a pronounced theta rhythm with highly similar features.


N-alkanes v. ytterbium/faecal index as two methods for estimating herbage intake of dairy cows fed on diets differing in the herbage: maize silage ratio and feeding level.
The aim of this study was to compare the n-alkanes and the ytterbium (Yb)/faecal index techniques as two methods for estimating the herbage intake of dairy cows fed indoors on different herbage : supplement ratios and feeding levels. The supplement was a mixture of maize silage and soyabean meal (ratio of 87 : 13 on a dry matter (DM) basis). In all, four treatments were studied. The herbage : supplement ratio in the diet was 25 : 75, 50 : 50, 75 : 25 and 50 : 50 for treatments 1, 2, 3 and 4, respectively. Animals were offered for treatments 1, 2 and 3, 100% of ad libitum intake measured before the experiment and 70% of ad libitum intake for treatment 4. Cows were fed herbage in the morning and supplement in the evening. A total of six lactating Holstein dairy cows were used in a 4 × 4 Latin square with four 14-day periods. Herbage and supplement intakes, faecal output (FO), in vivo organic matter (OM) digestibility and faecal recovery of markers were measured on the last 5 days of each period. Intake was estimated with the two methods and from two faecal sampling techniques, that is, total faecal collection v. grab sampling during milking. Mean herbage intake as fed, or estimated from n-alkanes or from the Yb/faecal index was 7.7, 8.1 and 10.2 kg DM, respectively. The mean prediction error, expressed as a fraction of actual herbage intake, was 0.10 and 0.50 for the n-alkanes and Yb/faecal index methods, respectively. The n-alkanes method clearly showed much better accuracy than the Yb/faecal index method for estimating intake, irrespective of the faecal sampling method, herbage : silage proportion or feeding level. For the n-alkanes method, herbage intake was slightly overestimated (7%) when herbage proportion in the diet was high, due to a ratio of faecal C33 : C32 recovery >1. The high bias for the Yb/faecal index was due to the cumulative effect of overestimation of FO (mean recovery of Yb = 0.92) and underestimation of the diet indigestible fraction (-8%). Between-treatment variations of FO were on average well estimated by Yb. Between-treatment variations of OM digestibility estimated using the faecal index technique were lower than those observed in vivo. It is concluded that intake of grazing dairy cows receiving high levels of maize silage supplement should be estimated using the n-alkanes method.


Continual skin peeling syndrome. An electron microscopic study.
We encountered a patient with continual skin peeling syndrome, a rare disorder in which generalized, noninflammatory exfoliation of the stratum corneum occurs. Although scaling occurred spontaneously in our patient, he was also able to manually peel sheets of skin without bleeding or pain. Histologically, there was separation of corneocytes above the granular cell layer. Ultrastructural examination revealed an unusual type of intracellular cleavage, in which the plasma membrane of the "peeling" cell remained firmly adherent to the underlying cell while the upper part of the cell exfoliated. Unique intercellular electron-dense globular deposits were localized to the stratum corneum.


Effect of endothelin-1 on human platelet shape change: reversal of activation by naftidrofuryl.
Naftidrofuryl (Praxilene; NAF) significantly improves claudication distance in patients with peripheral vascular disease (PVD). Endothelin-1 (ET-1) is a powerful endogenous vasoconstrictor and the circulating levels of ET-1 are elevated in patients with vascular disease. Platelet rich plasma (PRP) was prepared from healthy volunteers. NAF at concentrations similar to therapeutic levels (3.5-14 micromol/l), inhibited (P < 0.02) platelet activation (as indicated by a fall in median platelet volume, MPV) induced by ET-1 (0.4 micromol/l) alone. NAF also inhibited (P <0.0001) shape change (PSC; an early phase of platelet activation, characterised by an increase in MPV) induced by ET-1 (0.4 micromol/l) in combination with ADP (0.05-0.15 micromol/l) or serotonin (0.03-0.13 micromol/ l). We assessed the effect of ET(A) (BQ123, 50 nmol/l) or ET(B) (BQ788, 50 nmol/l) receptor antagonists on PSC induced by ET-1 alone. Both antagonists significantly inhibited PSC. We conclude that ET-1 activates human platelets. Both ET(A) and ET(B) receptors probably contribute to this response by a complex mechanism that requires further elucidation. NAF antagonises the action of ET-1 on human platelets. These actions may contribute to the beneficial effects of NAF in PVD.


Propranolol for recalcitrant ulcerated hemangioma of infancy.
Hemangioma of infancy (HOI) is a common, self-resolving, benign tumor that frequently requires no intervention. Ulceration is one of the main complications of HOI, and active treatment is usually required to manage pain, potential scarring, and occasionally bleeding and infection. Since the discovery that oral propranolol is an effective treatment for complicated HOI, it has been replacing systemic corticosteroids as first-line therapy, and early recommendations for treatment protocols have been devised. We report the successful treatment of six children with recalcitrant ulcerated HOI using propranolol. In all cases, complete, rapid healing occurred. Two children experienced recurrence of ulceration after ceasing propranolol, but this was rapidly controlled by restarting treatment. No adverse effects were encountered. We propose that ulceration in HOI is an indication for propranolol treatment if more conservative therapies have failed.