Longitudinal application of the medical outcomes study 36-item short-form health survey with not-in-treatment crack-cocaine users.
The Medical Outcomes Study SF-36 has been used infrequently with substance abusers. The objectives of this study were to use the SF-36 to assess changes in the health status of crack-cocaine users over time, to determine the characteristics of users who change, to assess the influence of frequency of crack use on SF-36 health status, and to determine the utility of the SF-36 with crack users. A multilevel model was used to analyze data that were collected every 6 months over a 2-year period. The sample consisted of 439 not-in-treatment crack-cocaine users. The SF-36 was administered at baseline and follow-up interviews. Data on the frequency of crack-cocaine use were also collected. On average, SF-36 health status remained fairly static during the study although statistically significant increases in scores occurred on the social functioning and role-emotional subscales. Variation in subjects' health trajectories occurred but was not, for the most part, explained by individual characteristics. Frequency of crack use was negatively related to scores on the physical functioning, social functioning, and mental health subscales. Most subscales did not reflect what are commonly recognized to be the health-compromising effects of cocaine use, raising questions about the use of the SF-36 with illicit drug users. Changes in crack users' SF-36 health status were uncommon. Frequency of crack use was negatively associated with health status. Several SF-36 subscales may be helpful in studies involving illicit drug users in which limited assessments of health status are needed.


Common cancers share familial susceptibility: implications for cancer genetics and counselling.
It has been proposed that cancer is more common in some families than in others, but the hypothesis lacks population level support. We use a novel approach by studying any cancers in large three-generation families and thus are able to find risks even though penetrance is low. Individuals in the nation-wide Swedish Family-Cancer Database were organised in three generations and the relative risk (RR) of cancer was calculated to the persons in the third generation by the numbers of patients with cancer in generations 1, 2 and 3. The RRs for any cancer in generation 3 increased by the numbers of affected relatives, reaching 1.61 when at least seven relatives were diagnosed. The median patient had two affected relatives, and 7.0% had five or more affected relatives with an RR of 1.46, which translated to an absolute risk of 21.5% compared with 14.7% in population by age 65 years. For prostate cancer, the RR was 2.85 with four or more affected family members with any cancer, and it increased to 14.42 with four or more concordant cancers in family members. RRs for prostate cancer were approximately equal (2.70 vs 2.85) if a man had one relative with prostate cancer or four or more relatives diagnosed with any cancer. A strong family history of cancer, regardless of tumour type, increases cancer risk of family members and calls for mechanistic explanations. Our data provide tools for counselling of patients with cancer with both low and high familiar risks.


The effects of monocyte-conditioned medium and interleukin 1 on the synthesis of collagenous and non-collagenous proteins by mouse bone and human bone cells in vitro.
Cultured adherent human mononuclear cells produce factor(s) which stimulate the release of calcium from new-born mouse calvaria in organ culture. This stimulation of bone resorption is accompanied by an inhibition of the incorporation of [3H]proline into collagen which is independent of increased prostaglandin production by the bone. When human osteoblast-like cells are treated with conditioned medium from human mononuclear cells, collagen accounts for a decreased proportion of the protein synthesised. This effect on matrix synthesis is not accompanied by an inhibitory action of the monocyte-conditioned medium preparations on net cell proliferation. In human osteoblast-like cell cultures, partially purified human interleukin 1 also inhibits the production of the bone-specific protein osteocalcin in a dose-dependent fashion. These observations are consistent with the hypothesis that products of human monocytes similar to, or identical with, human interleukin 1 may be important regulators of bone metabolism and may contribute to the bone loss seen in diseases such as chronic rheumatoid arthritis.


Pax3 and regulation of the melanocyte-specific tyrosinase-related protein-1 promoter.
Previous work has established that the melanocyte-specific tyrosinase-related protein-1 (TRP-1) promoter is regulated positively by the microphthalmia-associated transcription factor Mitf, acting through the conserved M box and negatively by the T-box factor Tbx2, which can bind two "melanocyte-specific elements" termed the MSEu and MSEi. Both the MSEu and MSEi, which share a 6-base pair GTGTGA consensus, are also recognized by a previously unidentified melanocyte-specific factor, MSF. Here we show using a combination of DNA binding assays, proteolytic clipping, and anti-Pax3 antibodies that MSF is indistinguishable from Pax3, a paired homeodomain transcription factor implicated genetically in melanocyte development and the regulation of the Mitf promoter. Consistent with Pax3 being able to bind the TRP-1 promoter, Pax3 is expressed in melanocytes and melanomas, and TRP-1 promoter activity is up-regulated by Pax3. The results identify a novel role for Pax3 in the expression of TRP-1, and the potential role of Pax3 in the melanocyte lineage is discussed.


Inhibiting amyloid-β cytotoxicity through its interaction with the cell surface receptor LilrB2 by structure-based design.
Inhibiting the interaction between amyloid-β (Aβ) and a neuronal cell surface receptor, LilrB2, has been suggested as a potential route for treating Alzheimer's disease. Supporting this approach, Alzheimer's-like symptoms are reduced in mouse models following genetic depletion of the LilrB2 homologue. In its pathogenic, oligomeric state, Aβ binds to LilrB2, triggering a pathway to synaptic loss. Here we identify the LilrB2 binding moieties of Aβ (16KLVFFA21) and identify its binding site on LilrB2 from a crystal structure of LilrB2 immunoglobulin domains D1D2 complexed to small molecules that mimic phenylalanine residues. In this structure, we observed two pockets that can accommodate the phenylalanine side chains of KLVFFA. These pockets were confirmed to be 16KLVFFA21 binding sites by mutagenesis. Rosetta docking revealed a plausible geometry for the Aβ-LilrB2 complex and assisted with the structure-guided selection of small molecule inhibitors. These molecules inhibit Aβ-LilrB2 interactions in vitro and on the cell surface and reduce Aβ cytotoxicity, which suggests these inhibitors are potential therapeutic leads against Alzheimer's disease.


Localized changes in apoplastic and cytoplasmic pH are associated with root hair development in Arabidopsis thaliana.
Morphogenesis in plants is characterized by highly regulated cell enlargement. However, the mechanisms controlling and localizing regions of growth remain essentially unknown. Root hair formation involves the induction of a localized cell expansion in the lateral wall of a root epidermal cell. This expanded region then enters a second phase of localized growth called tip growth. Root hair formation therefore provides a model in which to study the cellular events involved in regulating localized growth in plants. Confocal ratio imaging of the pH of the cell wall revealed an acidification at the root hair initiation site. This acidification was present from the first morphological indications of localized growth, but not before, and was maintained to the point where the process of root hair initiation ceased and tip growth began. Preventing the wall acidification with pH buffers arrested the initiation process but growth resumed when the wall was returned to an acidic pH. Cytoplasmic pH was found to be elevated from approximately 7.3 to 7. 7 at the initiation site, and this elevation coincided with the acidification of the wall. Preventing the localized increase in cytoplasmic pH with 10 mM butyrate however did not inhibit either the wall acidification or the initiation process. In contrast, there was no detectable gradient in pH associated with the apex of tip growing root hairs, but both elevated apoplastic pH and butyrate treatment irreversibly inhibited the tip growth process. Thus the processes of tip growth and initiation of root hairs show differences in their pH requirements. These results highlight the role of localized control of apoplastic pH in the control of cell architecture and morphogenesis in plants.


The Medical and Economic Burden of Narcolepsy: Implications for Managed Care.
The neurologic disorder narcolepsy results from dysregulation of the sleep-wake cycle and is primarily characterized by chronic, severely excessive daytime sleepiness and cataplexy, an emotionally induced muscle weakness. The prevalence of narcolepsy is approximately 0.05%, and onset generally occurs during the first 2 decades of life. Narcolepsy is believed to be an autoimmune disorder with destruction of hypocretin-producing neurons in the lateral hypothalamus. To provide an enhanced understanding of narcolepsy and establish the need for early diagnosis and rapid initiation of effective treatment for patients with narcolepsy. Narcolepsy reduces daily functioning and is associated with a substantial medical and economic burden, with many patients being on full disability. The annual direct medical costs are approximately 2-fold higher in patients with narcolepsy than in matched controls without this condition ($11,702 vs $5261, respectively; P <.0001). Further contributing to the overall burden is a lack of recognition of the signs and symptoms of narcolepsy and an absence of easily measurable biomarkers, resulting in a diagnostic delay that often exceeds 10 years and may be associated with misdiagnosis and inappropriate resource utilization. Because narcolepsy generally has an onset in childhood or in adolescence, is often misdiagnosed, has no known cure, and requires lifelong treatment, it is an important disease from a managed care perspective. Clinical features, as well as objective testing, should be used to ensure the timely diagnosis and treatment of patients with narcolepsy. Policies for the diagnosis and treatment of narcolepsy should be based on the current treatment guidelines, but they should also encourage shared decisions between clinicians and patients to allow for individualized diagnostic and treatment choices, as suggested in best practice recommendations.


Similarities between pandemics and cancer in growth and risk models.
Cancer and pandemics are leading causes of death globally, with severe socioeconomic repercussions. To better understand these repercussions, we investigate similarities between pandemics and cancer and describe the limited growth in number of infections or cancer cells, using mathematical models. For a pandemic, the analysis shows that in most cases, the initial fast growth is followed by a slower decay in the recovery phase. The risk of infection increases due to the airborne virus contact crossing a risk-threshold. For cancers caused by carcinogens, the increasing risk with age and absorbed dose of toxins that cross a risk-threshold, may lead to the disease onset. The time scales are different for both causes of death: years for cancer development and days to weeks for contact with airborne viruses. Contamination by viruses is on a time scale of seconds or minutes. The risk-threshold to get ill and the number-threshold in cancer cells or viruses, may explain the large variability in the outcome. The number of infected persons per day is better represented in log-lin plots instead of the conventional lin-lin plots. Differences in therapies are discussed. Our mathematical investigation between cancer and pandemics reveals a multifactorial correlation between both fragilities and brings us one step closer to understand, timely predict and ultimately diminish the socioeconomic hurdle of both cancer and pandemics.


Type 2 diabetes among Circassians in Israel.
The Muslim Circassians in Israel represent a unique ethnic community, distinct from Jews and Arabs. This endogamous group has a limited genetic variability that allows studying risk factors associated with type 2 diabetes. To estimate the prevalence of type 2 diabetes among Israeli Circassians and its correlation to obesity and genetic susceptibility. Israeli Circassian women (n = 450) and men (n = 289) older than 35 were included in the study. They were classified as having or not having diabetes, and their risk factors (including hypertension, body mass index, family history of diabetes) and laboratory tests, were examined retrospectively. The age-adjusted prevalence of diabetes among the 739 participants was 12% (men 14.6%, women 10.7%). It was higher among those with BMI > 30 than in those with lower BMI and a family history of diabetes without high BMI. But the risk of diabetes with BMI > 30 plus a family history was three times higher than when these factors were missing (odds ratio 2.96, 95% confidence interval 1.30-6.6). Multivariate analysis, however, found familial history of diabetes to be the strongest risk factor, independent of obesity (OR 2.47, 95% CI 1.45-4.20). The results yielded by this homogeneous Circassian population, sharing the same environmental influences and having an endogamous pattern of marriage, suggest a role of genetic risk factors for diabetes. Israeli Circassians are suitable for additional genetic studies that may lead to the identification of susceptibility genes for type 2 diabetes.


Standoff two-color quantum ghost imaging through turbulence.
Recently, a two-color quantum ghost imaging configuration was proposed by Karmakar et al. [Phys. Rev. A81, 033845 (2010)]. By illuminating an object located far away from the source and detector, with a signal beam of long wavelength to avoid absorption of short wavelengths in the atmosphere while a reference beam of short wavelength is detected locally, this imaging configuration can be appropriate for standoff sensing. In practice, the signal beam must propagate through atmosphere in the presence of serious turbulence. We analyzed theoretically the performance of this ghost imaging configuration through turbulence. Based on the Gaussian state source model and extended Huygens-Fresnel integral, a formula is derived to depict the ghost image formed through turbulence of a standoff reflective object. Numerical calculations are also given according to the formula. The results show that the image quality will be degraded by the turbulence, but the resolution can be improved by means of optimizing the wavelengths of the reference and signal beams even when the turbulence is very serious.


How humans integrate the prospects of pain and reward during choice.
The maxim "no pain, no gain" summarizes scenarios in which an action leading to reward also entails a cost. Although we know a substantial amount about how the brain represents pain and reward separately, we know little about how they are integrated during goal-directed behavior. Two theoretical models might account for the integration of reward and pain. An additive model specifies that the disutility of costs is summed linearly with the utility of benefits, whereas an interactive model suggests that cost and benefit utilities interact so that the sensitivity to benefits is attenuated as costs become increasingly aversive. Using a novel task that required integration of physical pain and monetary reward, we examined the mechanism underlying cost-benefit integration in humans. We provide evidence in support of an interactive model in behavioral choice. Using functional neuroimaging, we identify a neural signature for this interaction such that, when the consequences of actions embody a mixture of reward and pain, there is an attenuation of a predictive reward signal in both ventral anterior cingulate cortex and ventral striatum. We conclude that these regions subserve integration of action costs and benefits in humans, a finding that suggests a cross-species similarity in neural substrates that implement this function and illuminates mechanisms that underlie altered decision making under aversive conditions.


Cow, farm, and herd management factors in the dry period associated with raised somatic cell counts in early lactation.
This study investigated cow characteristics, farm facilities, and herd management strategies during the dry period to examine their joint influence on somatic cell counts (SCC) in early lactation. Data from 52 commercial dairy farms throughout England and Wales were collected over a 2-yr period. For the purpose of analysis, cows were separated into those housed for the dry period (6,419 cow-dry periods) and those at pasture (7,425 cow-dry periods). Bayesian multilevel models were specified with 2 response variables: ln SCC (continuous) and SCC >199,000 cells/mL (binary), both within 30 d of calving. Cow factors associated with an increased SCC after calving were parity, an SCC >199,000 cells/mL in the 60 d before drying off, increasing milk yield 0 to 30 d before drying off, and reduced DIM after calving at the time of SCC estimation. Herd management factors associated with an increased SCC after calving included procedures at drying off, aspects of bedding management, stocking density, and method of pasture grazing. Posterior predictions were used for model assessment, and these indicated that model fit was generally good. The research demonstrated that specific dry-period management strategies have an important influence on SCC in early lactation.


A modular approach for multifunctional polymersomes with controlled adhesive properties.
The bottom-up approach in synthetic biology involves the engineering of synthetic cells by designing biological and chemical building blocks, which can be combined in order to mimic cellular functions. The first step for mimicking a living cell is the design of an appropriate compartment featuring a multifunctional membrane. This is of particular interest since it allows for the selective attachment of different groups or molecules to the membrane. In this context, we report on a modular approach for polymeric vesicles, so-called polymersomes, with a multifunctional surface, namely hydroxyl, alkyne and acrylate groups. We demonstrate that the surface of the polymersome can be functionalized to facilitate imaging, via fluorescent dyes, or to improve the specific adhesion to surfaces by using a biotin functionalization. This generally applicable multifunctionality allows for the covalent integration of various molecules in the membrane of a synthetic cell.


Ascorbic acid metabolites are involved in intraocular pressure control in the general population.
Elevated intraocular pressure (IOP) is an important risk factor for glaucoma. Mechanisms involved in its homeostasis are not well understood, but associations between metabolic factors and IOP have been reported. To investigate the relationship between levels of circulating metabolites and IOP, we performed a metabolome-wide association using a machine learning algorithm, and then employing Mendelian Randomization models to further explore the strength and directionality of effect of the metabolites on IOP. We show that O-methylascorbate, a circulating Vitamin C metabolite, has a significant IOP-lowering effect, consistent with previous knowledge of the anti-hypertensive and anti-oxidative role of ascorbate compounds. These results enhance understanding of IOP control and may potentially benefit future IOP treatment and reduce vision loss from glaucoma.


Allocentric memory impaired and egocentric memory intact as assessed by virtual reality in recent-onset schizophrenia.
Present evidence suggests that schizophrenia is associated with explicit memory deficits, whereas implicit memory seems to be largely preserved. Virtual reality studies on declarative allocentric memory in schizophrenia are rare, and studies on implicit egocentric memory in schizophrenia are lacking. However, virtual realities have a major advantage for the assessment of spatial navigation and memory formation, as computer-simulated first-person environments can simulate navigation in a large-scale space. Twenty-five subjects with recent-onset schizophrenia were compared with 25 healthy matched control subjects on two virtual reality tasks affording the navigation and learning of a virtual park (allocentric memory) and a virtual maze (egocentric memory). Compared with control subjects, schizophrenia subjects were significantly impaired in learning the virtual park. However, schizophrenia subjects were as able as control subjects to learn the virtual maze. Stronger disorganized symptoms of schizophrenia subjects were significantly related to more errors on the virtual maze. It is concluded that egocentric spatial learning adds to the many other implicit cognitive skills being largely preserved in schizophrenia. Possibly, the more global neural network supporting egocentric spatial learning is less affected than the declarative hippocampal memory system in early stages of schizophrenia and may offer opportunities for compensation in the presence of focal deficits.


Molecular Basis of Disease Resistance in Banana Progenitor Musa balbisiana against Xanthomonas campestris pv. musacearum.
Banana Xanthomonas wilt disease, caused by Xanthomonas campestris pv. musacearum (Xcm), is a major threat to banana production in east Africa. All cultivated varieties of banana are susceptible to Xcm and only the progenitor species Musa balbisiana was found to be resistant. The molecular basis of susceptibility and resistance of banana genotypes to Xcm is currently unknown. Transcriptome analysis of disease resistant genotype Musa balbisiana and highly susceptible banana cultivar Pisang Awak challenged with Xcm was performed to understand the disease response. The number of differentially expressed genes (DEGs) was higher in Musa balbisiana in comparison to Pisang Awak. Genes associated with response to biotic stress were up-regulated in Musa balbisiana. The DEGs were further mapped to the biotic stress pathways. Our results suggested activation of both PAMP-triggered basal defense and disease resistance (R) protein-mediated defense in Musa balbisiana as early response to Xcm infection. This study reports the first comparative transcriptome profile of the susceptible and resistant genotype of banana during early infection with Xcm and provide insights on the defense mechanism in Musa balbisiana, which can be used for genetic improvement of commonly cultivated banana varieties.


Biomechanics of the elbow and forearm.
The elbow joint functions as a link between the shoulder and wrist providing an exceptional amount of stability and motion. The elbow joint should not be considered a simple hinge joint; rather there are important out-of-plane motions that affect implant design and surgical reconstruction of the elbow. Forearm rotation, which occurs as the radius wraps around the ulna, guided by the soft tissues at the wrist and elbow joints, also contributes to upper extremity mobility. Force transmission in the forearm is a complex interaction of the radius, ulna, and interosseous membrane. Forces at the wrist affect the transmission of force through the forearm to the elbow joint. During some activities, forces at the elbow joint can reach several times body weight. These motion, stability, and force interactions allow for the forearm's function. It should be recognized that an event at the elbow joint affects the forearm and the wrist, and conversely injury or disease at the wrist joint can affect the elbow.


Increased tonic cannabinoid CB1R activity and brain region-specific desensitization of CB1R Gi/o signaling axis in mice with global genetic knockout of monoacylglycerol lipase.
In mammalian brain, monoacylglycerol lipase (MAGL) is the primary enzyme responsible for terminating signaling function of the endocannabinoid 2-arachidonoylglycerol (2-AG). Previous in vivo studies with mice indicate that both genetic and chronic pharmacological inactivation of MAGL result in 8-30-fold increase of 2-AG concentration in the brain, causing desensitization and downregulation of cannabinoid CB1 receptor (CB1R) activity, leading to functional and behavioral tolerance. However, direct evidence for reduced CB1R activity in the brain is lacking. In this study, we used functional autoradiography to assess basal and agonist-stimulated CB1R-dependent Gi/o protein activity in multiple brain regions of MAGL-KO mice in comparison to their wild-type (WT) littermates. In addition, the role of endogenous cannabinoids in basal CB1R signaling was assessed after comprehensive pharmacological blockade of 2-AG hydrolysis by determining the contents of endocannabinoids (eCBs) in WT and MAGL-KO brain tissues by LC/MS/MS technology. To show whether lack of MAGL cause compensatory alterations in the serine hydrolase activity, we compared serine hydrolase pattern of WT and MAGL-KO using activity-based protein profiling. Consistent with studies using chronic pharmacological MAGL inactivation in vivo, we observed a statistically significant decrease of CB1R-Gi/o signaling in most of the studied brain regions. In MAGL-KO brain sections, elevated 2-AG levels were mirrored to heightened basal CB1R-dependent Gi/o-activity, as well as, dampened agonist-evoked responses in several brain regions. The non-selective serine hydrolase inhibitor methylarachidonoylfluorophosphonate (MAFP) was able to significantly elevate 2-AG levels in brain sections of MAGL-KO mice, indicating that additional serine hydrolases possess 2-AG hydrolytic activity in MAGL-KO brain sections.


Development of Sendai virus vectors and their potential applications in gene therapy and regenerative medicine.
Gene delivery/expression vectors have been used as fundamental technologies in gene therapy since the 1980s. These technologies are also being applied in regenerative medicine as tools to reprogram cell genomes to a pluripotent state and to other cell lineages. Rapid progress in these new research areas and expectations for their translation into clinical applications have facilitated the development of more sophisticated gene delivery/expression technologies. Since its isolation in 1953 in Japan, Sendai virus (SeV) has been widely used as a research tool in cell biology and in industry, but the application of SeV as a recombinant viral vector has been investigated only recently. Recombinant SeV vectors have various unique characteristics, such as low pathogenicity, powerful capacity for gene expression and a wide host range. In addition, the cytoplasmic gene expression mediated by this vector is advantageous for applications, in that chromosomal integration of exogenous genes can be undesirable. In this review, we introduce a brief historical background on the development of recombinant SeV vectors and describe their current applications in gene therapy. We also describe the application of SeV vectors in advanced nuclear reprogramming and introduce a defective and persistent SeV vector (SeVdp) optimized for such reprogramming.


Dynamic prediction of risk of death using history of cancer recurrences in joint frailty models.
Evaluating the prognosis of patients according to their demographic, biological, or disease characteristics is a major issue, as it may be used for guiding treatment decisions. In cancer studies, typically, more than one endpoint can be observed before death. Patients may undergo several types of events, such as local recurrences and distant metastases, with death as the terminal event. Accuracy of clinical decisions may be improved when the history of these different events is considered. Thus, it may be useful to dynamically predict patients' risk of death using recurrence history. As previously applied within the framework of joint models for longitudinal and time to event data, we propose a dynamic prediction tool based on joint frailty models. Joint modeling accounts for the dependence between recurrent events and death, by the introduction of a random effect shared by the two processes. We estimate the probability of death between the prediction time t and a horizon t + w, conditional on information available at time t. Prediction can be updated with the occurrence of a new event. We proposed and compared three prediction settings, taking into account three different information levels. The proposed tools are applied to patients diagnosed with a primary invasive breast cancer and treated with breast-conserving surgery, followed for more than 10 years in a French comprehensive cancer center.


Impacts of ocean acidification on early life-history stages and settlement of the coral-eating sea star Acanthaster planci.
Coral reefs are marine biodiversity hotspots, but their existence is threatened by global change and local pressures such as land-runoff and overfishing. Population explosions of coral-eating crown of thorns sea stars (COTS) are a major contributor to recent decline in coral cover on the Great Barrier Reef. Here, we investigate how projected near-future ocean acidification (OA) conditions can affect early life history stages of COTS, by investigating important milestones including sperm motility, fertilisation rates, and larval development and settlement. OA (increased pCO2 to 900-1200 µatm pCO2) significantly reduced sperm motility and, to a lesser extent, velocity, which strongly reduced fertilization rates at environmentally relevant sperm concentrations. Normal development of 10 d old larvae was significantly lower under elevated pCO2 but larval size was not significantly different between treatments. Settlement of COTS larvae was significantly reduced on crustose coralline algae (known settlement inducers of COTS) that had been exposed to OA conditions for 85 d prior to settlement assays. Effect size analyses illustrated that reduced settlement may be the largest bottleneck for overall juvenile production. Results indicate that reductions in fertilisation and settlement success alone would reduce COTS population replenishment by over 50%. However, it is unlikely that this effect is sufficient to provide respite for corals from other negative anthropogenic impacts and direct stress from OA and warming on corals.


beta-Lactamase production by intestinal spirochaetes.
beta-Lactamase production was demonstrated in four of nineteen strains of intestinal spirochaetes isolated from human subjects. The enzyme was preferentially active against penicillins and was inhibited by clavulanic acid; it was membrane bound and non-inducible. No plasmids were detected in the intestinal spirochaetes and the beta-lactamase-production characteristic was not transferable to non-producing strains.


Preparation and Evaluation of Lipid-Based Liquid Crystalline Formulation of Fenofibrate.
Many drugs have poor water solubility and so the oral delivery of such drugs is usually associated with limitation of low bioavailability and lack of dose proportionality. Lipid-based liquid crystal (LC) systems are excellent potential formulations for increasing dissolution and bioavailability of drugs. The aim of the present study was to formulate lipid-based LC containing fenofibrate (FFB) as a hydrophobic drug. The studied variables included lipid and stabilizer concentrations and the type of stabilizer. The LC formation was identified by the polarized optical microscopic method. The effects of variables on formulation characteristics such as particle size, drug release, and rheological behavior were evaluated. The results showed that the prepared formulations had the particle size between 42 and 503 nm. The drug release profiles showed that FFB had the continuous release from the formulations and the highest dissolution efficiency was seen in formulation prepared by 1.5% of glyceryl monostearate and 0.5% of Pluronic F127 as the stabilizer. The change of stabilizer type from colloidal silica to Pluronic F127 increased the drug release, significantly. In the most formulations of FFB LCs, the DE% was more than the pure drug, and therefore, it seems that the liquid crystalline formulations can be effective for enhancing drug release. Furthermore, drug release rate depended on the stabilizer type so that the presence of colloidal silica caused slower drug release compared to Pluronic F127.


Evaluation of Effects of Radiofrequency Ablation of Ex vivo Liver Using the 1-Fr Wire Electrode.
Percutaneous and intraoperative radiofrequency ablation (RFA) has become a valued tool in the management of primary and secondary hepatic lesions. A recent FDA-approved endoscopic ultrasound (EUS)-guided RFA probe now offers promise to help manage such lesions. However, there are no data to determine the ideal power setting and duration of ablation needed to effectively treat hepatic masses. The aim of the study was to evaluate the macroscopic zone of hepatic injury for EUS-RFA using a variety of settings within a fresh porcine hepatic specimen. RFA was performed using the Habib EUS-RFA needle (EMcision Ltd, London, UK) which is a 1-Fr wire (0.33 mm, 0.013 inch) with a working length of 190 cm. A step by step approach to deliver radiofrequency energy at 5, 10, 15, 20, and 50 W of power and 10, 30, 60, 90, 120, and 300 seconds, respectively, was followed. Macroscopic and microscopic findings of the ablation zone were evaluated at each setting. The maximal zone (diameter, 8.2±0.14 mm; length, 20.85±0.21 mm) of coagulative necrosis was achieved using an ablation power of 10 W for duration of 90 seconds. Notably, increased power settings resulted in an unexpected and diminished effect on tissue destruction. Our findings support the use of 10 W power for 90 seconds for maximum ablation in the liver. Our data also provide initial guidance and alternate settings to be considered when performing EUS-RFA to adjust the ablation power and duration to match the lesion size, shape, and risk of injury to adjacent structures.


Preconditioning triggered by carbon monoxide (CO) provides neuronal protection following perinatal hypoxia-ischemia.
Perinatal hypoxia-ischemia is a major cause of acute mortality in newborns and cognitive and motor impairments in children. Cerebral hypoxia-ischemia leads to excitotoxicity and necrotic and apoptotic cell death, in which mitochondria play a major role. Increased resistance against major damage can be achieved by preconditioning triggered by subtle insults. CO, a toxic molecule that is also generated endogenously, may have a role in preconditioning as low doses can protect against inflammation and apoptosis. In this study, the role of CO-induced preconditioning on neurons was addressed in vitro and in vivo. The effect of 1 h of CO treatment on neuronal death (plasmatic membrane permeabilization and chromatin condensation) and bcl-2 expression was studied in cerebellar granule cells undergoing to glutamate-induced apoptosis. CO's role was studied in vivo in the Rice-Vannucci model of neonatal hypoxia-ischemia (common carotid artery ligature +75 min at 8% oxygen). Apoptotic cells, assessed by Nissl staining were counted with a stereological approach and cleaved caspase 3-positive profiles in the hippocampus were assessed. Apoptotic hallmarks were analyzed in hippocampal extracts by Western Blot. CO inhibited excitotoxicity-induced cell death and increased Bcl-2 mRNA in primary cultures of neurons. In vivo, CO prevented hypoxia-ischemia induced apoptosis in the hippocampus, limited cytochrome c released from mitochondria and reduced activation of caspase-3. Still, Bcl-2 protein levels were higher in hippocampus of CO pre-treated rat pups. Our results show that CO preconditioning elicits a molecular cascade that limits neuronal apoptosis. This could represent an innovative therapeutic strategy for high-risk cerebral hypoxia-ischemia patients, in particular neonates.


Fibrinogen Replacement Therapy for Traumatic Coagulopathy: Does the Fibrinogen Source Matter?
Fibrinogen is the first coagulation protein to reach critically low levels during traumatic haemorrhage. There have been no differential effects on clinical outcomes between the two main sources of fibrinogen replacement: cryoprecipitate and fibrinogen concentrate (Fg-C). However, the constituents of these sources are very different. The aim of this study was to determine whether these give rise to any differences in clot stability that may occur during trauma haemorrhage. Fibrinogen deficient plasma (FDP) was spiked with fibrinogen from cryoprecipitate or Fg-C. A panel of coagulation factors, rotational thromboelastography (ROTEM), thrombin generation (TG), clot lysis and confocal microscopy were performed to measure clot strength and stability. Increasing concentrations of fibrinogen from Fg-C or cryoprecipitate added to FDP strongly correlated with Clauss fibrinogen, demonstrating good recovery of fibrinogen (r2 = 0.99). A marked increase in Factor VIII, XIII and α2-antiplasmin was observed in cryoprecipitate (p < 0.05). Increasing concentrations of fibrinogen from both sources were strongly correlated with ROTEM parameters (r2 = 0.78-0.98). Cryoprecipitate therapy improved TG potential, increased fibrinolytic resistance and formed more homogeneous fibrin clots, compared to Fg-C. In summary, our data indicate that cryoprecipitate may be a superior source of fibrinogen to successfully control bleeding in trauma coagulopathy. However, these different products require evaluation in a clinical setting.


Applying the ROBINS-I tool to natural experiments: an example from public health.
A new tool to assess Risk of Bias In Non-randomised Studies of Interventions (ROBINS-I) was published in Autumn 2016. ROBINS-I uses the Cochrane-approved risk of bias (RoB) approach and focusses on internal validity. As such, ROBINS-I represents an important development for those conducting systematic reviews which include non-randomised studies (NRS), including public health researchers. We aimed to establish the applicability of ROBINS-I using a group of NRS which have evaluated non-clinical public health natural experiments. Five researchers, all experienced in critical appraisal of non-randomised studies, used ROBINS-I to independently assess risk of bias in five studies which had assessed the health impacts of a domestic energy efficiency intervention. ROBINS-I assessments for each study were entered into a database and checked for consensus across the group. Group discussions were used to identify reasons underpinning lack of consensus for specific questions and bias domains. ROBINS-I helped to systematically articulate sources of bias in NRS. However, the lack of consensus in assessments for all seven bias domains raised questions about ROBINS-I's reliability and applicability for natural experiment studies. The two RoB domains with least consensus were selection (Domain 2) and performance (Domain 4). Underlying the lack of consensus were difficulties in applying an intention to treat or per protocol effect of interest to the studies. This was linked to difficulties in determining whether the intervention status was classified retrospectively at follow-up, i.e. post hoc. The overall risk of bias ranged from moderate to critical; this was most closely linked to the assessment of confounders. The ROBINS-I tool is a conceptually rigorous tool which focusses on risk of bias due to the counterfactual. Difficulties in applying ROBINS-I may be due to poor design and reporting of evaluations of natural experiments. While the quality of reporting may improve in the future, improved guidance on applying ROBINS-I is needed to enable existing evidence from natural experiments to be assessed appropriately and consistently. We hope future refinements to ROBINS-I will address some of the issues raised here to allow wider use of the tool.


Mass-Action Expressions of Ion Exchange Applied to Ca, H, K, and Mg Sorption on Isolated Cells Walls of Leaves from Brassica oleracea.
The cation exchange properties of cell walls isolated from collard (Bassica oleracea var acephala D.C.) leaves were investigated. Cation sorption on cell walls was described by mass-action expressions of ion exchange, rather than by the traditional Donnan equilibrium. The mass-action expressions enable the selectivity of the wall for one cation over another to be determined unambiguously from ion exchange isotherms. We found that: (a) the cation composition of the wall varied as a function of the solution cation concentration, solution cation composition, and pH in a way predicted by mass action; (b) the affinity of the wall for divalent cations increased as the equivalent fraction of divalent cation on the wall increased, and as the concentration of divalent cations in solution increased; (c) the selectivity of the wall for any metal cation pair was not altered by the concentration of H(+) in solution or on the wall; (d) H(+) sorption on the wall may be treated as a cation exchange reaction making it possible to calculate the relative affinity of the wall for metal cation pairs from H(+)-metal (Me) titration curves; and (e) the relative affinity of the wall for the cations we studied was: H(+) >> (K(+) >/= Ca(2+)) > Mg(2+). A cation-exchange model including surface complexes is consistent with observed cation selectivity. We conclude that metal cations interact with the wall to minimize or eliminate long-range electrostatic interactions and suggest that this may be due to the formation of site-specific cation-wall surface complexes.


Scheimpflug and high-resolution magnetic resonance imaging of the anterior segment: a comparative study.
High-resolution imaging with a camera system built on the Scheimpflug principle has been used to characterize the geometry of the anterior segment of the adult human eye as a function of aging and accommodative state but is critically dependent on algorithms for correction of distortion. High-resolution magnetic resonance imaging (MRI), in contrast, provides lower-resolution information about the adult eye but is undistorted. To test the accuracy of the Scheimpflug correction methods used by Cook and Koretz [J. Opt. Soc. Am. A 15, 1473 (1998)]; [Appl. Opt. 30, 2088 (1991)], data on anterior chamber and segment lengths, as well as lens thickness and anterior and posterior curvatures, were compared with corresponding MRI data for adults aged 18-50 at 0 diopter accommodation. Excellent statistical agreement was found between the MRI and the Scheimpflug data sets with the exception of the posterior lens radius of curvature, which is less well defined than the other measurements in the Scheimpflug images. The considerable agreement between data obtained with MR and Scheimpflug imaging, two different yet complementary in vivo imaging techniques, validates the Scheimpflug correction algorithms of Cook and Koretz and suggests the capability of directly integrating information from both. A third, equivalent, data set obtained with a Scheimpflug-style camera system differs considerably from both Scheimpflug and MRI results in magnitude and age dependence, with negative implications for this alternative method and its correction procedures.


Identification of molecular species of glycerophospholipids and sphingomyelin using electrospray mass spectrometry.
This paper describes the use of positive and negative ion electrospray mass spectrometry (MS) and MS/MS (tandem mass spectrometry) to identify glycerophospholipid and ceramide headgroups and their alkyl, alkenyl and acyl constituents. Molecular ion adducts were the primary products formed by positive ionization, occurring as [M+H]+, [M+Na]+, [M+K]+, [M+formate]+, or [M+acetate]+, depending upon the class of glycerophospholipid and the presence or absence of these ionization-promoting species. Similar (negatively charged) ions corresponding to the loss of the groups listed above were formed in negative ion MS. Positive ion electrospray MS/MS provides information on the nature of the headgroup, with the formation of an ion corresponding to the headgroup itself, or the loss of the headgroup from the molecular ion H+ or Na+ adduct. Acyl constituents are identified during negative ion MS/MS from the formation of their RCOO- ions. The nature of alkyl or alkenyl substituents in glycerophosphoethanolamine (PE) molecular species can be identified from residual ions following the loss of ethanolamine plus loss of the acyl moiety in the sn-2 position, and cyclization of a phosphate oxygen with C-2 of glycerol. In glycerophosphoinositol (PI) species, it appears that an RCO- ion is formed during negative ion MS/MS, possibly to steric interference from the bulky phosphoinositol headgroup that prevents cyclization (and subsequent stabilization) of the ion described for PE species. Positive and negative ion electrospray MS spectra for molecular species of commercial preparations of PE, PI, phosphatidylserine (PS), glycerophosphocholine (PC) and sphingomyelin (SM) produced similar profiles. For phospholipids occurring as Na+ adducts, concentrations above ca. 1 ng/microliter produced significant quantities of both [M+H]+ and [M+Na]+ ions for those molecular species present in the largest quantities, complicating interpretation of the spectra. Complete profiles of molecular species were obtained from as little as 10 picograms of material. Major components of PE were identified from 0.1 picogram total lipid. Using single ion monitoring of the Na+ adduct of beta-acetyl-gamma-O-hexadecyl L-alpha-phosphatidylcholine, 10 femtograms of material was detected. A mixture of 1 nanogram each of PE, PI, PS, and PC was readily resolved into individual molecular species, with little apparent loss of resolution or preferential ionization. Electrospray MS did not provide information on the position (sn-1 or sn-2) of fatty acids, and was not capable of differentiating in all instances between alkyl-acyl and alkenyl-acyl substituents without prior separation of these lipid subclasses.(ABSTRACT TRUNCATED AT 400 WORDS)


Intravaginal brachytherapy in FIGO stage I low-risk endometrial cancer: a controlled randomized study.
The purpose of the study was to compare postoperative vaginal irradiation with surgery alone in low-risk International Federation of Gynecology and Obstetrics (FIGO) stage IA-IB endometrial carcinoma. The study was a prospective, randomized trial of 645 evaluable low-risk endometrial carcinoma patients from 6 European gynecologic cancer centers. All tumors were in FIGO stage IA-IB, of endometrioid histological type, and FIGO grade 1-2. High-dose-rate afterloading equipments (iridium [Ir] 192 or cobalt [co] 60) were used at 5 centers, and low-dose-rate (LDR) afterloading equipment (cesium [Cs] 137) at 1 center. Perspex vaginal applicators or ovoids were normally used, and the dose was specified at 5 mm from the surface of the applicator. Three to 6 fractions (3.0-8.0 Gy) were given, and the overall treatment time was 4 to 15 days. A total of 319 patients were treated with surgery plus vaginal irradiation (treatment group), and 326 patients with surgery alone (control group).Twenty-six recurrences (4.0%) were recorded in the complete series. The locoregional recurrence rate was 2.6%, whereas distant metastases occurred in 1.4%. The rate of vaginal recurrences was 1.2% in the treatment group versus 3.1% in the control group. The difference was not statistically significant (P = 0.114). Side effects were few and mild (grade 1-2). Dysuria, frequency, and incontinence were slightly more common after vaginal irradiation (2.8% vs 0.6%, respectively). Late intestinal problems were few and similar in the 2 groups. The conclusions were that the impact of postoperative brachytherapy on even the locoregional recurrence rate seems to be limited in patients with low-risk endometrial carcinoma. The overall recurrence rate and survival were similar in the 2 groups.


Temporally Distinct Regulation of Pathways Contributing to Cardiac Proteostasis During the Acute and Recovery Phases of Sepsis.
Cardiac dysfunction is a common manifestation of sepsis and is associated with early increases in inflammation and decreases in myocardial protein synthesis. However, little is known regarding the molecular mechanisms regulating protein homeostasis during the recovery phase after the removal of the septic nidus. Therefore, the purpose of this study was to investigate diverse signal transduction pathways that regulate myocardial protein synthesis and degradation. Adult male C57BL/6 mice were used to identify potential mechanisms mediating the acute (24 h) effect of cecal ligation and puncture as well as long-term changes that manifest during the chronic (10 days) recovery phase. Sepsis acutely decreased cardiac protein synthesis that was associated with reduced phosphorylation of S6K1/S6 but not 4E-BP1. Sepsis also decreased proteasome activity, although with no change in MuRF1 and atrogin-1 mRNA expression. Sepsis acutely increased apoptosis (increased caspase-3 and PARP cleavage), autophagosome formation (increased LC3B-II), and canonical inflammasome activity (increased NLRP3, TMS1, cleaved caspase-1). In contrast, during the recovery phase, independent of a difference in food consumption, global protein synthesis was increased, the early repression in proteasome activity was restored to basal levels, whereas stimulation of apoptosis, autophagosome formation, and the canonical inflammasome pathway had abated. However, during recovery there was a selective stimulation of the noncanonical inflammasome pathway as evidenced by activation of caspase-11 with cleavage of Gasdermin D. These data demonstrate a temporally distinct homeostatic shift in the cardiac proteostatic response to acute infection and recovery.


A facile supramolecular approach to fabricate multifunctional upconversion nanoparticles as a versatile platform for drug loading, in vivo delivery and tumor imaging.
Multifunctional upconversion nanoparticles (UCNPs) that can be implemented in theranostic applications are particularly attractive scaffolds for precise drug delivery. However, most of the current methods for drug formulation are technically complicated, thereby impeding their use in the clinic. Here, we report on the preparation of a lipophilic cytotoxic prodrug-integrated and polyethylene glycol (PEG)-cloaked UCNPs scaffold through a facile one-pot supramolecular approach. By choosing 7-ethyl-10-hydroxycamptothecin (SN38)-derived prodrug 1 as a model chemotherapeutic, we show that this lipophilic prodrug can be feasibly self-assembled onto the surface of UCNPs, which are cooperatively solubilized by PEGylated phospholipids. The resulting SN38 prodrug 1-encapsulated UCNPs (designated 1@pUCNPs) produce a stable colloidal system in aqueous solution, making it suitable for intravenous injection. The SN38 drug loading capacity in pUCNPs is as high as ∼12.3 wt%, and a sustained drug release profile is observed, indicating that the drug payloads can be transported to targeted tumor sites via the enhanced permeability and retention (EPR) effect. Upconversion luminescence (UCL) imaging, including in vivo and ex vivo imaging, suggests that the drug-loaded pUCNPs remain stable in tumors over a long time and preferentially accumulate in tumors presumably via the EPR effect. Furthermore, the 1@pUCNPs show superior therapeutic outcomes compared with the clinically approved SN38 prodrug CPT-11 in the Bcap-37 mouse model of breast cancer. Collectively, our results demonstrate that pUCNPs facilely constructed in a one-pot self-assembly manner may be used as a versatile platform, enabling synchronous in vivo delivery of poorly water-soluble drugs and tumor imaging.


Alcohol drinking patterns differentially affect central adiposity as measured by abdominal height in women and men.
Alcohol drinking in light-to-moderate amounts has been associated with reduced coronary heart disease (CHD) risk. However, there is evidence that the way people consume alcohol (drinking pattern) may affect risk. Central adiposity, a known CHD risk factor may be one mechanism in the pathway between alcohol consumption and CHD risk. Our study examined whether various drinking patterns differentially affect fat distribution, particularly abdominal fat in women and men. In a randomly selected population-based cohort (n = 2343), 35-79 y old, we assessed drinking pattern as reported for the past 30 d, including beverage type and amount, frequency of consumption, percentage of time drinking while eating and number of drinks consumed/drinking day. Central adiposity was determined using an abdominal caliper to measure supine height of the abdomen. Current drinkers tended to have smaller abdominal heights than nondrinkers (women, P < 0.0001; men, P = 0.0559). For drinking pattern, frequency was inversely associated, but drinking intensity (drinks/drinking day) was positively associated with central adiposity in women (P trend for frequency, 0.0007; intensity, 0.0010) and men (P trend for frequency, 0.0005; intensity, 0.0004), even when age, education, physical activity, smoking status and amount of alcohol (g) were included in the models. When frequency and intensity were considered together, daily drinkers of <1 drink/drinking day had the smallest mean abdominal height measures with the largest measures in less than weekly drinkers who consumed 4 or more drinks/drinking day. These results support the hypothesis that drinking pattern affects the distribution of body fat, an important CHD risk factor.


Pre-treatment psoas major volume is a predictor of poor prognosis for patients with epithelial ovarian cancer.
Low skeletal muscle mass (sarcopenia) is an important prognostic risk factor for the outcome of a variety of cancer types. The current study investigated whether skeletal muscle area (SMA), psoas area (PA) and psoas major volume (PV) are associated with progression-free survival (PFS) and overall survival (OS) in patients with epithelial ovarian cancer (OC). A total of 92 OC patients were enrolled in the present study. Pre-treatment with SMA and PA was assessed using computed tomography (CT) and PV was calculated using a three-dimensional-CT (3D-CT). The clinical factors associated with sarcopenia and prognosis were retrospectively evaluated. For all patients, the median PFS and OS were 19 and 32 months, respectively. Patients exhibiting lower PV (<195.6 cm3) had significantly poorer PFS and OS compared with patients exhibiting higher PV (≥195.6 cm3; P=0.018 and P=0.006), while those with low SMA (<92.92 cm2) had significantly worse OS than patients with higher SMA (≥92.92 cm2; P=0.030). PV was also demonstrated to be superior to SMA and PA in prognosis prediction. PV by 3D-CT can serve as an indicator of poor prognosis in patients with OC.


Association of child care providers breastfeeding support with breastfeeding duration at 6 months.
Many lactating mothers participate in the workforce and have their infants cared for outside of their home, yet little is known about their child care providers' (CCPs') support of breastfeeding. This study examines the association between CCPs' breastfeeding support as reported by mothers at 3 months and mother's breastfeeding at 6 months. Infant Feeding Practices Study II, a longitudinal study, followed mothers of infants via mail questionnaires almost monthly from late pregnancy throughout the first year. This study consisted of 183 mothers who breastfed and had their infant in child care at 3 months and answered 5 questions regarding CCPs' supports. Total number of CCPs' support was a summary of responses to individual items and categorized into 3 levels (0-2, 3-4, or 5 total supports). Multiple logistic regressions examined how each breastfeeding support and total number were associated with breastfeeding at 6 months. Breastfeeding at 6 months was significantly associated with CCP support to feed expressed breast milk (AOR = 4.55; 95% CI = 1.09, 18.95) and allow mothers to breastfeed at the child care place before or after work (AOR = 6.23; 95% CI = 1.33, 29.16). Compared to mothers who reported fewer than 3 total supports, mothers who reported 5 supports were 3 times as likely to be breastfeeding at 6 months (AOR = 3.00, 95% CI = 1.11, 8.13). Our findings suggest that CCPs' breastfeeding support at 3 months, particularly feeding expressed breast milk and allowing mothers to breastfeed before or after work, may help mothers maintain breastfeeding at 6 months.


Antiproliferative effects of 1,25-dihydroxyvitamin D3 on primary cultures of human prostatic cells.
Cultures of adult human prostatic epithelial and fibroblastic cells were established from normal, benign hyperplastic, and malignant tissues. Vitamin D receptors were detected by ligand binding of [3H]1,25-dihydroxyvitamin D3 [1,25(OH)2D3] in cytosolic extracts prepared from all types of cell cultures as well as from fresh prostatic tissues. Vitamin D receptor transcripts were demonstrated by Northern blot analysis. 1,25-(OH)2D3 inhibited the growth of epithelial cells with half-maximal inhibition at approximately 1 nM. The growth of fibroblasts was also inhibited by 1,25(OH)2D3 but to a lesser extent. This is consistent with the apparently lower level of vitamin D receptors in fibroblasts compared to epithelial cells determined by ligand binding and Northern analysis of RNA transcripts. The growth inhibition of epithelial cells by 1,25(OH)2D3 was irreversible even after a short 2-h exposure, but morphology and keratin expression were not appreciably altered by long-term exposure to the hormone. A physiological role for 1,25(OH)2D3 in the prostate is postulated, and the inhibitory effect of 1,25(OH)2D3 on cancer-derived prostate cells may provide a basis for new preventive or therapeutic strategies.


Effect of minimal listing criteria on waiting list registration for liver transplantation: a process-outcome analysis.
To determine the level of association between minimal listing criteria (MLC) recognition and outcomes associated with waiting list registration for liver transplantation (LT). A total of 147 patients and 201 patients were identified as first-time referrals for LT evaluation between January 1, 1997, and November 30, 1997 (cohort A), and December 1,1997, and December 31, 1998 (cohort B), respectively. Relevant demographic and clinical information was abstracted from medical records. Minimal listing criteria were defined as a Child-Turcotte-Pugh (CTP) score of 7 or higher. Patient age, sex, hepatic disease etiology, and mean CTP scores were similar between cohorts A and B. However, the proportion of registered patients in cohort B with CTP scores of 7 or higher increased significantly after formal MLC recognition (96% vs 82% for cohort A; P=.001). In cohort A, waiting list registration was based on patient age, male sex, nonalcohol-related hepatic disease, and a CTP score of 7 or higher in the absence of formal MLC. The rate of first-time patient referral was also increased in cohort B vs cohort A after formal MLC recognition (80% vs 69%, respectively; P=.002) despite similar clinical characteristics. Although the number of patients with a CTP score of 10 or higher was greater in cohort B vs cohort A, the proportion of patients with advanced end-stage liver disease was similar (29% vs 26%, respectively; P=.72). The explicit recognition of MLC was strongly associated with improvements in appropriate waiting list registration for LT.


Signaling from G protein-coupled receptors to ERK5/Big MAPK 1 involves Galpha q and Galpha 12/13 families of heterotrimeric G proteins. Evidence for the existence of a novel Ras AND Rho-independent pathway.
The regulation of gene expression by cell surface receptors often involves the stimulation of signaling pathways including one or more members of the MAPK superfamily of serine-threonine kinases. Upon their activation in the cytosol, MAPKs can translocate to the nucleus and affect the activity of a variety of transcription factors. Recently, it has been observed that a novel member of the MAPK superfamily, ERK5, can be potently activated by transforming G protein-coupled receptors (GPCRs) and that ERK5 participates in the regulation of c-jun expression through the activation of MEF2 transcription factors. How cell surface receptors, including GPCRs, stimulate ERK5 is still poorly understood. In this study, we have used transiently transfected COS-7 cells to begin delineating the biochemical route linking GPCRs to ERK5. We show that receptors that can couple to the G(q) and G(12/13) families of heterotrimeric G proteins, m1 and thrombin receptors, respectively, but not those coupled to G(i), such as m2 receptors, are able to regulate the activity of ERK5. To investigate which heterotrimeric G proteins signal to ERK5, we used a chimeric system by which Galpha(q)- and Galpha(13)-mediated signaling pathways can be conditionally activated upon ligand stimulation. Using this system, as well as the expression of activated forms of G protein subunits, we show that the Galpha(q) and Galpha(12/13) families of heterotrimeric G proteins, but not the Galpha(i), Galpha(s), and betagamma subunits, are able to regulate ERK5. Furthermore, we provide evidence that the stimulation of ERK5 by GPCRs involves a novel signaling pathway, which is distinct from those regulated by Ras and Rho GTPases.


Meeting highlights: international expert consensus on the primary therapy of early breast cancer 2005.
The ninth St Gallen (Switzerland) expert consensus meeting in January 2005 made a fundamental change in the algorithm for selection of adjuvant systemic therapy for early breast cancer. Rather than the earlier approach commencing with risk assessment, the Panel affirmed that the first consideration was endocrine responsiveness. Three categories were acknowledged: endocrine responsive, endocrine non-responsive and tumors of uncertain endocrine responsiveness. The three categories were further divided according to menopausal status. Only then did the Panel divide patients into low-, intermediate- and high-risk categories. It agreed that axillary lymph node involvement did not automatically define high risk. Intermediate risk included both node-negative disease (if some features of the primary tumor indicated elevated risk) and patients with one to three involved lymph nodes without additional high-risk features such as HER 2/neu gene overexpression. The Panel recommended that patients be offered chemotherapy for endocrine non-responsive disease; endocrine therapy as the primary therapy for endocrine responsive disease, adding chemotherapy for some intermediate- and all high-risk groups in this category; and both chemotherapy and endocrine therapy for all patients in the uncertain endocrine response category except those in the low-risk group.


Compartmentalization of HIV-1 in the central nervous system: role of the choroid plexus.
To determine the genetic and phenotypic composition of HIV-1 found in the choroid plexus (CPx) and its relationship to virus in the brain and peripheral lymphoid tissue. Phenotypic and molecular comparisons of HIV-1 found in autopsy brain, CPx, and spleen tissues. HIV-1 was co-cultured from matched postmortem brain (basal ganglia), CPx, and spleen tissues of AIDS patients with and without HIV-associated encephalitis and dementia. Viral phenotypes were determined by infection of monocyte-derived macrophages, MT-2 or co-receptor-specific cell lines. Viral env and pol sequences were determined from genomic DNA isolated directly from tissues or co-cultures, and phylogenetic comparisons were performed. CCR5-utilization was the most prevalent viral tropism found in all tissues, although spleen isolates also displayed CXCR4 usage. Viruses isolated from CPx consisted of both peripheral and brain-like virus, but were more related phenotypically and genetically to those found in the brain. Mutations found in the pol gene that could confer drug resistance to brain and CPx isolates were similar to those found in the periphery. The CPx contained replication-competent virus that was most similar, although distinct, from that found in the brain. It also contained some viruses with high similarity to those of peripheral origin. Compartmentalization of viral env and pol sequences indicated that differential selective pressures exist in each tissue examined. These studies suggest that the CPx may provide an environment that promotes the evolution of drug-resistant strains with central nervous system tropism, although it is unlikely to be a reservoir for archival HIV-1 variants.


VASP regulates leukocyte infiltration, polarization, and vascular repair after ischemia.
In ischemic vascular diseases, leukocyte recruitment and polarization are crucial for revascularization and tissue repair. We investigated the role of vasodilator-stimulated phosphoprotein (VASP) in vascular repair. After hindlimb ischemia induction, blood flow recovery, angiogenesis, arteriogenesis, and leukocyte infiltration into ischemic muscles in VASP-/- mice were accelerated. VASP deficiency also elevated the polarization of the macrophages through increased signal transducer and activator of transcription (STAT) signaling, which augmented the release of chemokines, cytokines, and growth factors to promote leukocyte recruitment and vascular repair. Importantly, VASP deletion in bone marrow-derived cells was sufficient to mimic the increased blood flow recovery of global VASP-/- mice. In chemotaxis experiments, VASP-/- neutrophils/monocytes were significantly more responsive to M1-related chemokines than wild-type controls. Mechanistically, VASP formed complexes with the chemokine receptor CCR2 and β-arrestin-2, and CCR2 receptor internalization was significantly reduced in VASP-/- leukocytes. Our data indicate that VASP is a major regulator of leukocyte recruitment and polarization in postischemic revascularization and support a novel role of VASP in chemokine receptor trafficking.


When cells become a drug. Endothelial progenitor cells for cardiovascular therapy: aims and reality.
The recently disclosed plasticity properties of adult-derived stem cells, their ability to be reprogrammed by defined factors into pluripotent stem cells and the comprehension of "epi"-genetic mechanisms underlying stem cells differentiation process has opened unexpected avenues to attempt regeneration of tissues affected by degenerative disorders and prompted the birth of the new "regenerative medicine" concept. Regeneration of the vascular and myocardial tissues is considered a primary endpoint to limit the consequences of acute and chronic ischemic heart disorders. Cellular therapy of the ischemic heart has been attempted in more than 1000 patients worldwide and the results of the first meta-analysis studies have been recently made available. In several cases, the results did not fulfill the expectations. In fact, they unpredictably indicated modest, yet significant, clinical benefits in patients compared to the outstanding results using stem cells in animal models of ischemic heart and peripheral disease. Several interpretations have been raised to explain these discrepancies. These include lifestyle and risk factor-associated modifications of the stem cell biological activity, but also procedural problems in the translation of cells from bench to bedside. The present review will cover light and shaded areas in the cardiovascular cellular therapy field, and will discuss about recent advances and related patents designed to enhance efficiency of stem cell therapy in patients with cardiovascular disease. These advancements will be discussed in the light of the most advanced issues that have been introduced worldwide by Regulatory Agencies.


Epidural analgesia use as a marker for physician approach to birth: implications for maternal and newborn outcomes.
Understanding the association between caregiver belief systems and practice patterns is an emerging area of research. We hypothesized an association between a maternity caregiver's belief system and his or her behavior. The study objective was to determine if a family physician's overall approach to maternity care, as measured by average use of epidural analgesia, was associated with maternal and fetal outcomes. Retrospective analysis was conducted of the births of three cohorts of 1992 nulliparous, low-risk women attended by 96 family physicians within an 18-month period in the department of family practice at the largest maternity hospital in Canada. Cohorts were based on the physicians' mean use of epidural analgesia for the women. Family physicians attending fewer than 5 births were excluded. The main outcome measures, by physician epidural utilization cohort, were maternal/newborn morbidity, procedure rates, consultation rates, and length of stay. Family physicians were separated into cohorts based on their mean use of epidural analgesia at rates of: low, 0-30 percent (15 physicians, 263 births); medium, 31-50 percent (55 physicians, 1323 births); and high, 51-100 percent (26 physicians, 406 births). After adjustment for maternal age and race, patients of low versus high epidural users were admitted at a later state of cervical dilation (mean 4.0 vs 3.1 cm), received less electronic fetal monitoring (76.4 vs 87.2%) and oxytocin augmentation (12.2 vs 29.8%), sustained fewer malpositions (occiput posterior or transverse) (23.2 vs 34.2%), had fewer cesarean sections (14.0 vs 24.4%), less obstetric consultation (47.9 vs 63.8%), and fewer newborn special care admissions (7.2 vs 12.8%). In our setting, high use of epidural analgesia is a marker for a style of practice characterized by malpositions leading to dysfunctional labors and higher intervention rates leading, in turn, to excess maternal/newborn morbidity.


Dose-dependent relationship between oocyte cytoplasmic volume and transformation of sperm nuclei to metaphase chromosomes.
We have studied the chromosome condensation activity of mouse oocytes that have been inseminated during meiotic maturation. These oocytes remain unactivated, and in those penetrated by up to three or four sperm, each sperm nucleus is transformed, without prior development of a pronucleus, into metaphase chromosomes. However, those penetrated by more than four sperm never transform any of the nuclei into metaphase chromosomes (Clarke, H. J., and Y. Masui, 1986, J. Cell Biol. 102:1039-1046). We report here that, when the cytoplasmic volume of oocytes was doubled or tripled by cell fusion, up to five or eight sperm nuclei, respectively, could be transformed into metaphase chromosomes. Conversely, when the cytoplasmic volume was reduced by bisection of oocytes after the germinal vesicle (GV) had broken down, no more than two sperm could be transformed into metaphase chromosomes. Thus, the capacity of the oocyte cytoplasm to transform sperm nuclei to metaphase chromosomes was proportional to its volume. The contribution of the nucleoplasm of the GV and the cytoplasm outside the GV to the chromosome condensation activity was investigated by bisecting oocytes that contained a GV and then inseminating the nucleate and anucleate fragments. The anucleate fragments never induced sperm chromosome formation, indicating that GV nucleoplasm is required for this activity. In the nucleate fragments, the capacity to induce sperm chromosome formation was reduced as compared with whole oocytes, in spite of the fact that the fragments contained the entire GV nucleoplasm. This implies that non-GV cytoplasmic material also was required for chromosome condensation activity. When inseminated oocytes were incubated in the presence of puromycin, the sperm nuclei were transformed into interphase-like nuclei, but no metaphase chromosomes developed. However, when protein synthesis resumed, the interphase nuclei were transformed to metaphase chromosomes. These results suggest that the transformation of sperm nuclei to metaphase chromosomes in the cytoplasm of mouse oocytes requires both the nucleoplasm of the GV and non-GV cytoplasmic substances, including proteins synthesized during maturation.


Primary infertility in nuclear industry employees: report from the nuclear industry family study.
To determine whether men and women who are occupationally exposed to ionising radiation are at increased risk of primary infertility. A cross sectional analysis of reproductive outcome was reported in an occupational cohort of nuclear industry workers from the Atomic Energy Authority, Atomic Weapons Establishment, and British Nuclear Fuels in the United Kingdom. Data on employment and radiation monitoring supplied by employers were linked to data obtained from self administered questionnaires sent to all current employees and a sample of past workers. A total of 5353 men and 603 women aged 40 years or more at the time of survey whose first pregnancy, or attempt at pregnancy, had occurred after first employment in the nuclear industry were recruited to the study. Primary infertility was defined as consultation with a doctor about the failure to achieve at least one viable (24 weeks gestational age or more) pregnancy. 2.6% Of men and 3.7% of women reported primary infertility. For men, this proportion did not differ by amount of radiation monitored or by dose received. The prevalence of infertility was higher among monitored women than non-monitored women, but not significantly so and the numbers were too small to draw any firm conclusions. No support was found for a hypothesis linking exposure to low level ionising radiation among men with primary infertility. There was weak evidence of an association in women, but the relatively few monitored women prevented detailed examination of these data.


Therapeutic Benefits of Laughter in Mental Health: A Theoretical Review.
In modern society, fierce competition and socioeconomic interaction stress the quality of life, causing a negative influence on a person's mental health. Laughter is a positive sensation, and seems to be a useful and healthy way to overcome stress. Laughter therapy is a kind of cognitive-behavioral therapies that could make physical, psychological, and social relationships healthy, ultimately improving the quality of life. Laughter therapy, as a non-pharmacological, alternative treatment, has a positive effect on the mental health and the immune system. In addition, laughter therapy does not require specialized preparations, such as suitable facilities and equipment, and it is easily accessible and acceptable. For these reasons, the medical community has taken notice and attempted to include laughter therapy to more traditional therapies. Decreasing stress-making hormones found in the blood, laughter can mitigate the effects of stress. Laughter decreases serum levels of cortisol, epinephrine, growth hormone, and 3,4-dihydrophenylacetic acid (a major dopamine catabolite), indicating a reversal of the stress response. Depression is a disease, where neurotransmitters in the brain, such as norepinephrine, dopamine, and serotonin, are reduced, and there is something wrong in the mood control circuit of the brain. Laughter can alter dopamine and serotonin activity. Furthermore, endorphins secreted by laughter can help when people are uncomfortable or in a depressed mood. Laughter therapy is a noninvasive and non-pharmacological alternative treatment for stress and depression, representative cases that have a negative influence on mental health. In conclusion, laughter therapy is effective and scientifically supported as a single or adjuvant therapy.


Personality disorder diagnostic thresholds correspond to different levels of latent pathology.
To meet a diagnosis for a particular personality disorder (PD), among other requirements, a person must exhibit a specified minimum number of the features outlined for that PD, otherwise referred to as the diagnostic threshold. Despite many years of research on the DSM PDs, there is little empirical basis for the chosen thresholds. The present study used mechanisms of item response theory (IRT) to link the diagnostic thresholds of six PDs to their corresponding level of latent PD pathology. Consistent with our hypothesis, analyses of the data from 41,227 participants revealed that PD diagnostic thresholds corresponded to a wide range of latent pathology. For example, the diagnostic threshold for schizoid PD corresponded to 1.54 SDs of pathology whereas the threshold for dependent PD corresponded to 2.72 SDs of pathology. The current analyses have demonstrated for the first time that the latent pathology associated with each PD threshold varies widely, and thus has quantified what others accurately have speculated. Implications and considerations for the future directions of PD classification are discussed.


Aripiprazole (intramuscular) for psychosis-induced aggression or agitation (rapid tranquillisation).
People experiencing psychosis may become aggressive. Antipsychotics, such as aripiprazole in intramuscular form, can be used in such situations. To evaluate the effects of intramuscular aripiprazole in the treatment of psychosis-induced aggression or agitation (rapid tranquillisation). On 11 December 2014 and 11 April 2017, we searched the Cochrane Schizophrenia Group's Study-based Register of Trials which is based on regular searches of CINAHL, BIOSIS, AMED, Embase, PubMed, MEDLINE, PsycINFO, and registries of clinical trials. All randomised controlled trials (RCTs) that randomised people with psychosis-induced aggression or agitation to receive either intramuscular aripiprazole or another intramuscular intervention. We independently inspected citations and, where possible, abstracts, ordered papers and re-inspected and quality assessed these. We included studies that met our selection criteria. At least two review authors independently extracted data from the included studies. We chose a fixed-effect model. We analysed dichotomous data using risk ratio (RR) and the 95% confidence intervals (CI). We analysed continuous data using mean differences (MD) and their CIs. We assessed risk of bias for included studies and used GRADE to create 'Summary of findings' tables. Searching found 63 records referring to 21 possible trials. We could only include three studies, all completed over the last decade, with 885 participants, of which 707 were included for quantitative analyses in this systematic review. Due to limited comparisons, small size of trials and a paucity of investigated and reported 'pragmatic' outcomes, evidence was mostly graded as low or very low quality. No trials reported useful data for one of our primary outcomes of tranquil or asleep by 30 minutes. Economic outcomes were also not reported in the trials.When compared with placebo, fewer people in the aripiprazole group needed additional injections compared to the placebo group (2 RCTs, n = 382, RR 0.69, 95% CI 0.56 to 0.85, very low-quality evidence). Clinically important improvement in agitation at two hours favoured the aripiprazole group (2 RCTs, n = 382, RR 1.50, 95% CI 1.17 to 1.92, very low-quality evidence). The numbers of non-responders after the first injection also favoured aripiprazole (1 RCT, n = 263, RR 0.49, 95% CI 0.34 to 0.71, low-quality evidence). Although no effect was found, more people in the aripiprazole compared to the placebo group experienced adverse effects (1 RCT, n = 117, RR 1.51, 95% CI 0.93 to 2.46, very low-quality evidence).Aripiprazole required more injections compared to haloperidol (2 RCTs, n = 477, RR 1.28, 95% CI 1.00 to 1.63, very low-quality evidence), with no significant difference in agitation (2 RCTs, n = 477, RR 0.94, 95% CI 0.80 to 1.11, very low-quality evidence), and similar non-responders after first injection (1 RCT, n = 360, RR 1.18, 95% CI 0.78 to 1.79, low-quality evidence). Aripiprazole and haloperidol did not differ when taking into account the overall number of people that experienced at least one adverse effect (1 RCT, n = 113, RR 0.91, 95% CI 0.61 to 1.35, very low-quality evidence).Compared to aripiprazole, olanzapine was better at reducing agitation (1 RCT, n = 80, RR 0.77, 95% CI 0.60 to 0.99, low-quality evidence) and had a more favourable effect on global state change scores (1 RCT, n = 80, MD 0.58, 95% CI 0.01 to 1.15, low-quality evidence), both at two hours. No differences were found in terms of experiencing at least one adverse effect during the 24 hours after treatment (1 RCT, n = 80, RR 0.75, 95% CI 0.45 to 1.24, very low-quality evidence). However, participants allocated to aripiprazole experienced less somnolence (1 RCT, n = 80, RR 0.25, 95% CI 0.08 to 0.82, low-quality evidence). The available evidence is of poor quality but there is some evidence aripiprazole is effective compared to placebo and haloperidol, but not when compared to olanzapine. However, considering that evidence comes from only three studies, caution is required in generalising these results to real-world practice. This review firmly highlights the need for more high-quality trials on intramuscular aripiprazole in the management of people with acute aggression or agitation.


A 371-nucleotide region between the herpes simplex virus type 1 (HSV-1) LAT promoter and the 2-kilobase LAT is not essential for efficient spontaneous reactivation of latent HSV-1.
The herpes simplex virus type 1 (HSV-1) latency-associated transcript (LAT) gene is essential for efficient spontaneous reactivation of HSV-1 from latency. However, neither the mechanism by which LAT carries out this function nor the region of LAT responsible for this function in known. LAT is transcribed as an unstable 8.3-kb RNA that gives rise to a very stable 2-kb LAT RNA that is readily detected in latently infected sensory neurons. We show here that 371 of the 662 nucleotides located between the start of LAT transcription and the 5' end of the 2-kb LAT RNA do not appear to be essential for wild-type levels of spontaneous reactivation in the rabbit ocular model of HSV-1 latency. We deleted LAT nucleotides 76 to 447 from both copies of the LAT gene (one in each viral long repeat) to produce the mutant dLAT371. Rabbits were ocularly infected with dLAT371, and spontaneous reactivation was measured in comparison with the marker-rescued virus dLAT371R. Both dLAT371 and dLAT371R had spontaneous reactivation rates of approximately 13 to 14%. This was consistent with the parental McKrae wild-type virus (11.7%; P = 0.49) and significantly higher than the LAT transcription-negative mutant dLAT2903 (2.4%; P < 0.0001). Southern analysis confirmed that the spontaneously reactivated dLAT371 virus retained the deletion in both copies of LAT. Therefore, it appeared that the function of LAT involved in efficient spontaneous reactivation mapped outside the 371-nucleotide region deleted from the LAT gene of dLAT371.


Antibodies to acetylcholine receptor in parous women with myasthenia: evidence for immunization by fetal antigen.
The weakness in myasthenia gravis (MG) is mediated by autoantibodies against adult muscle acetylcholine receptors (AChR) at the neuromuscular junction; most of these antibodies also bind to fetal AChR, which is present in the thymus. In rare cases, babies of mothers with MG, or even of asymptomatic mothers, develop a severe developmental condition, arthrogryposis multiplex congenita, caused by antibodies that inhibit the ion channel function of the fetal AChR while not affecting the adult AChR. Here we show that these fetal AChR inhibitory antibodies are significantly more common in females sampled after pregnancy than in those who present before pregnancy, suggesting that they may be induced by the fetus. Moreover, we were able to clone high-affinity combinatorial Fab antibodies from thymic cells of two mothers with MG who had babies with arthrogryposis multiplex congenita. These Fabs were highly specific for fetal AChR and did not bind the main immunogenic region that is common to fetal and adult AChR. The Fabs show strong biases to VH3 heavy chains and to a single Vkappa1 light chain in one mother. Nevertheless, they each show extensive intraclonal diversification from a highly mutated consensus sequence, consistent with antigen-driven selection in successive steps. Collectively, our results suggest that, in some cases of MG, initial immunization against fetal AChR is followed by diversification and expansion of B cells in the thymus; maternal autoimmunity will result if the immune response spreads to the main immunogenic region and other epitopes common to fetal and adult AChR.


Relationships between state and trait anxiety inventory and alcohol use disorder identification test scores among Korean twins and families: the healthy twin study.
We explored heritabilities of the State and Trait Anxiety Inventory (STAI) and the Alcohol Use Disorders Identification Test (AUDIT), and associations including genetic and environmental correlations between the phenotypes among Korean twins and their families. We analyzed the data of 1,748 participants (835 men, 913 women, 656 individuals of monozygotic twins, 173 individuals of same-sexed dizygotic twins, 919 non-twin family members, age 30-79 years) from the Healthy Twin study. Heritabilities and bivariate analyses were assessed using the SOLAR package software. In the methods of generalized estimation equations, women in the 4th quartile of state and trait scores were 17% and 15%, respectively more likely to be hazardous alcohol users compared to women in the lower three quartiles (P < .05). However, there were no significant associations between these phenotypes in men. After adjusting for age and squared age, the heritability estimates were 0.26 in men and 0.34 in women for the state score; for the trait score, 0.35 in men and 0.31 in women; for the AUDIT score, 0.32 in men and 0.37 in women (P < .001). After adjusting for age and squared age, there was a significant genetic correlation between the trait score and the AUDIT score, and a significant non-genetic correlation between the state score and the AUDIT score in women, while there were no significant genetic or non-genetic correlations between these phenotypes in men. The STAI and AUDIT scores are heritable in Koreans and the relationships between these phenotypes may be inconsistent by sex.


Metabolic models to investigate energy limited anaerobic ecosystems.
Anaerobic wastewater treatment is shifting from a philosophy of solely pollutants removal to a philosophy of combined resource recovery and waste treatment. Simultaneous wastewater treatment with energy recovery in the form of energy rich products, brings renewed interest to non-methanogenic anaerobic bioprocesses such as the anaerobic production of hydrogen, ethanol, solvents, VFAs, bioplastics and even electricity from microbial fuel cells. The existing kinetic-based modelling approaches, widely used in aerobic and methanogenic wastewater treatment processes, do not seem adequate in investigating such energy limited microbial ecosystems. The great diversity of similar microbial species, which share many of the fermentative reaction pathways, makes quantify microbial groups very difficult and causes identifiability problems. A modelling approach based on the consideration of metabolic reaction networks instead of on separated microbial groups is suggested as an alternative to describe anaerobic microbial ecosystems and in particular for the prediction of product formation as a function of environmental conditions imposed. The limited number of existing relevant fermentative pathways in conjunction with the fact that anaerobic reactions proceed very close to thermodynamic equilibrium reduces the complexity of such approach and the degrees of freedom in terms of product formation fluxes. In addition, energy limitation in these anaerobic microbial ecosystems makes plausible that selective forces associated with energy further define the system activity by favouring those conversions/microorganisms which provide the most energy for growth under the conditions imposed.


Different fluorophore labeling strategies and designs affect millisecond kinetics of DNA hairpins.
Changes in molecular conformations are one of the major driving forces of complex biological processes. Many studies based on single-molecule techniques have shed light on conformational dynamics and contributed to a better understanding of living matter. In particular, single-molecule FRET experiments have revealed unprecedented information at various time scales varying from milliseconds to seconds. The choice and the attachment of fluorophores is a pivotal requirement for single-molecule FRET experiments. One particularly well-studied millisecond conformational change is the opening and closing of DNA hairpin structures. In this study, we addressed the influence of base- and terminal-labeled fluorophores as well as the fluorophore DNA interactions on the extracted kinetic information of the DNA hairpin. Gibbs free energies varied from ∆G0 = -3.6 kJ/mol to ∆G0 = -0.2 kJ/mol for the identical DNA hairpin modifying only the labeling scheme and design of the DNA sample. In general, the base-labeled DNA hairpin is significantly destabilized compared to the terminal-labeled DNA hairpin and fluorophore DNA interactions additionally stabilize the closed state of the DNA hairpin. Careful controls and variations of fluorophore attachment chemistry are essential for a mostly undisturbed measurement of the underlying energy landscape of biomolecules.


Chronic morphine treatment reduces recovery from opioid desensitization.
Tolerance and dependence result from long-term exposure to opioids, and there is growing evidence linking acute receptor desensitization to these more long-term processes. Receptor desensitization encompasses a series of events leading to the loss of receptor function and internalization. This study examines the onset and recovery from desensitization in locus ceruleus neurons recorded in brain slices taken from animals that have been chronically treated with morphine. After chronic morphine treatment, desensitization was altered as follows. First, the rate of desensitization was increased. Second, recovery from desensitization was always incomplete, even after a brief (1-2 min) exposure to agonist. This contrasts with experiments in controls in which recovery from desensitization, after a brief exposure to agonist, was complete within 25 min. Finally, morphine-6-beta-D-glucuronide, a metabolite of morphine that was ineffective at causing desensitization in controls, induced significant desensitization in slices from morphine-treated animals. When brain slices from controls were treated with inhibitors of PKC or monensin, agents known to compromise G-protein-coupled receptor resensitization, desensitization was increased, and recovery was significantly reduced. These results indicate that receptor resensitization maintains signaling during periods of intense and sustained stimulation. After chronic morphine treatment, desensitization is potentiated, and receptor resensitization is compromised.


The role of stromal cells in inflammatory bone loss.
Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation, local and systemic bone loss and a lack of compensatory bone repair. Fibroblast-like synoviocytes (FLS) are the most abundant cells of the stroma and a key population in autoimmune diseases such as RA. An increasing body of evidence suggests that these cells play not only an important role in chronic inflammation and synovial hyperplasia, but also impact bone remodelling. Under inflammatory conditions FLS release inflammatory cytokines, regulate bone destruction and formation and communicate with immune cells to control bone homeostasis. Other stromal cells, such as osteoblasts and terminally differentiated osteoblasts, termed osteocytes, are also involved in the regulation of bone homeostasis and are dysregulated during inflammation. This review highlights our current understanding of how stromal cells influence the balance between bone formation and bone destruction. Increasing our understanding of these processes is critical to enable the development of novel therapeutic strategies with which to treat bone loss in RA.


Emotional functioning of individuals with borderline personality traits in a nonclinical population.
Relatively few studies have evaluated the emotional functioning of individuals with borderline personality disorder traits, especially in nonclinical populations. This study therefore sought to understand further the emotional skills and subjective emotional experiences of adults with borderline traits in a community sample. Adult volunteers (N=523) were recruited from community and student populations, and borderline personality was determined via three self-report measures. Close to one in six respondents (17.2%) self-reported borderline personality traits above the threshold on the three instruments. Poor skills in managing and understanding emotion were characteristic of these individuals. They also possessed significantly poorer subjective perception of emotion, management of their own emotions and management of others' emotions, relative to the non-borderline personality controls. Skills in managing and understanding emotion and the subjective experience of managing one's own emotions were significant multivariate predictors of borderline personality trait status. We conclude that persons with borderline personality traits have pronounced deficits in emotional understanding and management. Interventions targeting these deficits are needed, given the high prevalence of borderline traits in the community.


Ultrafast pump-probe spectroscopy of neutral FenOm clusters (n, m < 16).
Neutral iron oxide clusters (FenOm, n, m ≤ 16) are produced in a laser vaporization source using O2 gas seeded in He. The neutral clusters are ionized with a sequence of femtosecond laser pulses and detected using time-of-flight mass spectrometry. Small clusters are confirmed to be most prominent in the stoichiometric (n = m) distribution, with m = n + 1 clusters observed above n = 4. Pump-probe spectroscopy is employed to study the dynamics of an electron transfer from an oxygen orbital to an iron nonbonding orbital of iron oxide clusters that is driven by absorption of a 400 nm photon. A bifurcation of the initial wavepacket occurs, where a femtosecond component is attributed to electron relaxation assisted through internuclear vibrational relaxation and high density of states, and a slow relaxation shows the formation of a bound excited state. The lifetime and relative ratio of the two pathways depend on both the cluster size and iron oxidation state. The femtosecond lifetime decreases with increased cluster size until a saturation timescale is achieved at n > 5. The relative population of the long-lived excited state decreases with cluster size and suggests that the excited electron remains on the Fe atom for >20 ps.


D-alanylation of lipoteichoic acids confers resistance to cationic peptides in group B streptococcus by increasing the cell wall density.
Cationic antimicrobial peptides (CAMPs) serve as the first line of defense of the innate immune system against invading microbial pathogens. Gram-positive bacteria can resist CAMPs by modifying their anionic teichoic acids (TAs) with D-alanine, but the exact mechanism of resistance is not fully understood. Here, we utilized various functional and biophysical approaches to investigate the interactions of the human pathogen Group B Streptococcus (GBS) with a series of CAMPs having different properties. The data reveal that: (i) D-alanylation of lipoteichoic acids (LTAs) enhance GBS resistance only to a subset of CAMPs and there is a direct correlation between resistance and CAMPs length and charge density; (ii) resistance due to reduced anionic charge of LTAs is not attributed to decreased amounts of bound peptides to the bacteria; and (iii) D-alanylation most probably alters the conformation of LTAs which results in increasing the cell wall density, as seen by Transmission Electron Microscopy, and reduces the penetration of CAMPs through the cell wall. Furthermore, Atomic Force Microscopy reveals increased surface rigidity of the cell wall of the wild-type GBS strain to more than 20-fold that of the dltA mutant. We propose that D-alanylation of LTAs confers protection against linear CAMPs mainly by decreasing the flexibility and permeability of the cell wall, rather than by reducing the electrostatic interactions of the peptide with the cell surface. Overall, our findings uncover an important protective role of the cell wall against CAMPs and extend our understanding of mechanisms of bacterial resistance.


Relationship between obesity and health problems in help-seeking military veterans.
UK Armed Forces (UK AF) veterans may be particularly vulnerable to obesity and its comorbid physical and mental health problems. To examine the relationship between body mass index (BMI), physical health problems, mental health disorders and sociodemographic characteristics in UK AF veterans engaged in psychological treatment. Information regarding veteran BMI, demographic characteristics, physical health conditions and mental health problems was collected and analysed using univariate and multivariate regression analyses. Rates of veteran obesity were also compared with the UK general population. Of the 384 help-seeking veterans, 37.5% (n=151) were overweight (BMI 26-30) and 35.5% (n=143) were obese. Obesity in help-seeking male veterans was two to four times higher than that of the general population of UK. Higher scores on measures of anger and common mental health problems were significantly associated with greater BMI. Problems with physical systemic functioning and mobility were also significantly associated with greater BMI. The results indicate that treatment-seeking UK AF veterans exhibit higher levels of obesity compared with the general population, and clinically significant physical and mental comorbidities. The findings highlight a need for mental health services to offer treatments that effectively integrate physical and mental healthcare in the treatment of people with mental health problems.


Personality characteristics of staff in elderly care-a cross-cultural comparison.
Communication and interaction between carers and residents in elderly and dementia care can be challenging and demanding. The carer's personality, one factor shaping this interaction, seems to have been neglected in the literature. This article looks at cross-cultural comparisons of staff in elderly and dementia care with individuals from the general population matched by age and gender. Compared to individuals in the general population, elderly and dementia care staff are usually slower tempered, more stoic and reflective, tolerant to monotony, and more systematic. They also have more optimistic attitudes in situations that might worry most people, and more confidence in social situations and in the face of danger and uncertainty.