Preventing Delirium and Promoting Long-Term Brain Health: A Clinical Trial Design for the Perioperative Cognitive Enhancement (PROTECT) Trial.
Perioperative neurocognitive disorders (PND), including postoperative delirium (POD), are common in older adults and, for many, precipitate functional decline and/or dementia. In this protocol, we describe a novel multidisciplinary, multicomponent perioperative intervention that seeks to prevent or reduce POD and associated cognitive decline. We will conduct a prospective, single-blind, pragmatic, randomized-controlled trial to compare our tailored multi-disciplinary perioperative pathway against current standard of care practices. We will recruit a total of 692 elective surgical patients aged 65 years or more and randomize them in a 1:1 design. Our perioperative intervention targets delirium risk reduction strategies by emphasizing the importance of early mobilization, nutrition, hydration, cognitive orientation, sensory aids, and avoiding polypharmacy. To promote healthy behavior change, we will provide a tailored psychoeducation program both pre- and postoperatively, focusing on cardiovascular and psychosocial risks for cognitive and functional decline. Our primary outcome is the incidence of any PND (encapsulating POD and mild or major postoperative neurocognitive disorder) at three months postoperative. Secondary outcomes include any incidence of POD or neurocognitive disorder at 12 months. A specialized delirium screening instrument, the Confusion Assessment Method (3D-CAM), and a neuropsychological test battery, will inform our primary and secondary outcomes. Delirium is a common and debilitating postoperative complication that contributes to the cognitive and functional decline of older adults. By adopting a multicomponent, multidisciplinary approach to perioperative delirium prevention, we seek to reduce the burden of delirium and subsequent dementia in older adults.


Oral immunotherapy in hen's egg-allergic children increases a hypo-proliferative subset of CD4+ T cells that could constitute a marker of tolerance achievement.
Food allergy affects a significant number of children and its prevalence, and persistence is undergoing an important increase in the last years. Specific oral tolerance induction (SOTI) is a promising therapy for food allergy. However, little is known about the immune mechanisms implicated in the desensitization to allergens. Our purpose was to study which immune parameters are modified during the process of tolerance achievement with the goal of identifying markers of tolerance induction. We performed an extensive immune analysis in 19 allergic children following SOTI with hen's egg before and after the immunotherapy. Changes in lymphocyte subpopulations and serum cytokines were identified in children with desensitization achievement. Sixteen children achieved complete tolerance to egg, and the immune analysis reveals that desensitization was accompanied in all the cases by a significant decrease in the percentage and absolute counts of effector-memory CD4+ T cells (T(EM) ) and a marked increase in the absolute counts of a subset of CD4(+) CD38(+) CD45RO(-) cells. Additionally, we also observed a marked reduction in the plasma levels of different Th1 and Th2 cytokines after tolerance achievement. Acquisition of tolerance in children after oral immunotherapy is accompanied by a decrease in the T(EM) population and the increase in a particular subset of CD4+ T cells with a hypo-proliferative and non-reactive phenotype. This hypo-proliferative subset of cells could constitute a marker of the development of oral tolerance, and the study of this subset could contribute to the better understanding of the immune responses in allergic subjects.


Regulation of ubiquitin-proteasome and autophagy pathways after acute LPS and epoxomicin administration in mice.
The ubiquitin-proteasome pathway (UPP) is a major protein degradation pathway that is activated during sepsis and has been proposed as a therapeutic target for preventing skeletal muscle loss due to cachexia. Although several studies have investigated the modulation of proteasome activity in response to LPS administration, none have characterized the overall UPP response to LPS administration in the fate of proteasome inhibition. Here, we determined the modulation pattern of the main key components of the UPP in the gastrocnemius (GAS) of mice during the acute phase of lipopolysaccharide (LPS)-mediated endotoxemia (7.5 mg/kg - 8 h) by measuring all three β1, β2 and β5 activites of the 20S and 26S proteasomes, the levels of steady state polyubiquitinated proteins, mRNA levels of muscle ligases, as well as signaling pathways regulating the UPP. Another goal was to assess the effects of administration of a specific proteasome inhibitor (epoxomicin, 0.5 mg/kg) on UPP response to sepsis. The acute phase of LPS-induced endotoxemia lowered GAS/body weight ratio and increased MuRF1 and MAFbx mRNA concomitantly to an activation of the pathways known to regulate their expression. Unexpectedly, we observed a decrease in all 20S and 26S proteasome activities measured in GAS, which might be related to oxidative stress, as oxidized proteins (carbonyl levels) increase with LPS. While significantly inhibiting 20S and 26S proteasome β5 activities in heart and liver, epoxomicin did not lower proteasome activity in GAS. However, the increase in mRNA expression of the muscle ligases MuRF1 and MAFbx were partially rescued without affecting the other investigated signaling pathways. LPS also strongly activated autophagy, which could explain the observed GAS atrophy with LPS-induced reduction of proteasome activity. Our results highlight an opposite regulation of UPP in the early hours of LPS-induced muscle atrophy by showing reduced proteasome activities and increased mRNA expression of muscle specific ligases. Furthermore, our data do not support any preventive effect of epoxomicin in muscle atrophy due to acute cachexia since proteasome activities are not further repressed.


Developing cultural competence in the baccalaureate nursing student: a population-based project with the Hutterites.
The purpose of this article is to articulate the process of educating baccalaureate students to apply culturally competent, population-based nursing care to the Hutterites. The Hutterites are a German dialect-speaking, ethnoreligious population. Campinha-Bacote's culturally competent model guided the student learning process. Groups of students developed culture-specific, population-based assessment models to identify the health-related needs of the Hutterites. One example of a Hutterite health assessment model that was derived from Dever's epidemiologic model and Purnell and Paulanka's model for cultural competence is presented. Priority health needs of Hutterite colonies are described, and examples of health programs are given. Recommendations for working with the Hutterite population as part of an undergraduate public health nursing course are put forth. The culturally congruent process of teaching and learning for nursing students can be applied to other health-related professions and adapted to other cultural contexts.


Clinical evaluation of polyacid-modified resin composite posterior restorations: one-year results.
The aim of this investigation was to evaluate the clinical performance of a new compomer restorative system, Dyract AP, placed in combination with Non-Rinse Conditioner and Prime & Bond NT in permanent posterior teeth. Fifty Class II and 41 Class I restorations were placed in 39 patients by 1 dentist. The restorations were evaluated directly, with modified US Public Health Service criteria, and indirectly, with color slides and polyvinyl siloxane impressions, at baseline and 6 months and 1 year after placement. Preoperative and 1-year postoperative bitewing radiographs were also taken. All 82 restorations available for 1-year evaluation were in situ. No postoperative sensitivity or pulpal problems were reported. Four Class II restorations (4.9%) failed because of partial fracture or recurrent caries. The percentages of Alfa score for each criterion were color match, 95.1%; marginal discoloration, 57.3%; marginal integrity, 35.4%; anatomic form, 98.8%; and surface texture, 91.5%. The average wear rate of Dyract AP was low (18.5 +/- 11.7 microns at 6 months and 35.7 +/- 13.6 microns at 12 months). The excellent handling characteristics, the good clinical performance, and the improved wear resistance suggest that this compomer will provide reliable direct tooth-colored restorations in stress-bearing areas.


Analytical toxicology of the semi-synthetic cannabinoid hexahydrocannabinol studied in human samples, pooled human liver S9 fraction, rat samples and drug products using HPLC-HRMS-MS.
Hexahydrocannabinol (HHC) is an emerging semi-synthetic cannabinoid, which is obtained from cyclization of cannabidiol and subsequent hydrogenation. As a potentially legal alternative of ∆9-tetrahydrocannabinol (∆9-THC), it is increasingly seized in the USA and Europe. The aims of this study were to investigate the metabolism of HHC in pooled human liver S9 fraction (pHLS9), rat and human samples. Additionally, a locally obtained low-THC cannabis product was investigated, which was advertised with an elevated concentration of HHC. Overall, HHC formed an 11-hydroxy (HO) metabolite, as well as a carboxy metabolite. While only the parent compound was detected in rat urine and feces, the hydroxy metabolite was additionally detected in pHLS9 and human plasma. The carboxy metabolite was only detectable in human plasma. The metabolism corresponded well to that of ∆9-THC, although glucuronidation and the formation of an 8-HO metabolite were not observed. Detectability of HHC and its carboxy metabolite in rat urine was investigated using gas chromatography-mass spectrometry, but neither the parent compound nor the metabolite were detectable. The investigated low-THC cannabis product appeared to be an actual cannabis product since, in addition to HHC, cannabinol, cannabidiol and ∆9-THC were detected after qualitative analysis. Estimation of its content revealed not only 30.6% of HHC but also 4% of ∆9-THC.


MEFIB-Index and MAST-Score in the assessment of hepatic decompensation in metabolic dysfunction-associated steatosis liver disease-Individual participant data meta-analyses.
There are limited data regarding the longitudinal association between MEFIB-Index (MRE combined with FIB-4) versus MAST-Score (MRI-aspartate aminotransferase) and hepatic decompensation. To examine the longitudinal association between MEFIB-Index versus MAST-Score in predicting hepatic decompensation in patients with metabolic dysfunction-associated steatotic liver disease (MASLD). This was a longitudinal, retrospective analysis of subjects from United States, Japan, and Turkey who underwent a baseline MRE and MRI-PDFF and were followed for hepatic decompensation. Cox-proportional hazard analyses were used to assess the association between MEFIB-Index versus MAST-Score with a composite primary outcome (hepatic decompensation) defined as ascites, hepatic encephalopathy, and varices needing treatment. This meta-analysis of individual participants (IPDMA) included 454 patients (58% women) with a mean (±SD) age of 56.0 (±13.5) years. The MEFIB-Index (MRE ≥3.3 kPa + FIB 4 ≥1.6) and MAST-Score (>0.242) were positive for 34% and 9% of the sample, respectively. At baseline, 23 patients met criteria for hepatic decompensation. Among 297 patients with available longitudinal data with a median (IQR) of 4.2 (5.0) years of follow-up, 25 incident cases met criteria for hepatic decompensation. A positive MEFIB-Index [HR = 49.22 (95% CI: 6.23-388.64, p < 0.001)] and a positive MAST-Score [HR = 3.86 (95% CI: 1.46-10.17, p < 0.001)] were statistically significant predictors of the incident hepatic decompensation. MEFIB-Index (c-statistic: 0.89, standard error (SE) = 0.02) was statistically superior to the MAST-Score (c-statistic: 0.81, SE = 0.03) (p < 0.0001) in predicting hepatic decompensation. A combination of MRI-based biomarker and blood tests, MEFIB-Index and MAST-Score can predict the risk of hepatic decompensation in patients with MASLD.


Entire calcaneus reconstruction with pedicled composite fibular growth plate flap in a pediatric patient.
Traumatic calcanectomy is a complicated rare condition that results in chronic pain and loss of limb function as the calcaneus is crucial for bearing weight. For an adult, possible treatment options are limited to below knee amputation, bone grafts, and 3D prosthesis; however, for the pediatric population, therapeutic data are scarce. In this case report, the authors describe a technique that provides the possibility for hindfoot reconstruction while maintaining growth potential without traumatizing other anatomic regions. A 5-year-old patient presented in the emergency department after he accidentally slipped under a lawnmower, which resulted in full calcaneal amputation with large bone and plantar soft tissue defects from the medial to lateral malleoli. Four days later, reconstructive surgery was performed using a composite vascularized pedicle growth plate flap. The flap was based on the retrograde anterior tibial artery and consisted of the peroneus longus muscle and fasciocutaneous perforator flap to maintain soft tissue coverage. The calcaneus was reconstructed by a double barrel fibular transplant localizing the growth plate in the distal part of the neocalcaneus. Full consolidation of the fibular barrels was achieved and the growth plate was still open at the 8-year follow-up. The fibular transplant has acquired a similar shape to a calcaneus with symmetric flattening and opposite side angulation. The patient's lower extremity functional score was 78/80 points. Reconstruction of an amputated calcaneal bone is rare and complicated, especially for pediatric patients. Authors demonstrate treatment with a complex vascularized flap for sustained growth and good functional outcome in the long term.


Infertility in the Global South: Raising awareness and generating insights for policy and practice.
Infertility is a highly prevalent reproductive health condition in the global South, which often has a devastating impact on the people concerned. Yet, thus far it hardly received any attention from policy makers, Non-Governmental Organizations (NGOs) or donors working in the field of Sexual and Reproductive Health and Rights (SRHR). For this reason we have set up a project to increase knowledge and awareness about infertility and childlessness among those stakeholders and organizations and to generate insight into (possible) interventions in this field. The project received a grant by Share-Net International (the Knowledge Platform in the field of SRHR, funded by the Dutch Ministry of Foreign Affairs) and is a unique collaboration between universities, fertility clinics, fertility support groups and the Walking Egg Foundation. The project consists of multimethods studies in Ghana and Kenya as well as dissemination workshops and meetings in these countries and the Netherlands. The first workshops in Kenya have already taken place with successful feedback from stakeholders. In this commentary we provide insight into the project and the main points and recommendations discussed in the Workshops in Kenya.


Novel EDTA-ligands containing an integral perylene bisimide (PBI) core as an optical reporter unit.
The synthesis, characterization and metal complexation of a new class of perylene bisimides (PBIs) as an integral part of ethylenediaminetetraacetic acid (EDTA) are reported. The simplest representative, namely derivative 1a, was synthesized both by a convergent as well as a direct approach while the elongated derivatives, 1b and 1c, were obtained only via a convergent synthetic pathway. All these new prototypes of water-soluble perylenes are bolaamphiphiles and were fully characterized by (1)H- and (13)C-NMR spectroscopy, matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry and IR spectroscopy. In order to acquaint ourselves with the behaviour in solution of our PBIs bearing dendritic wedges, the simplest derivative, 1a, was chosen and tested by means of UV/Vis and fluorescence spectroscopy as well as by zeta-potential measurements. A photoexcitation induced intramolecular photo-electron transfer (PET) can be observed in these molecules. Therefore their potential applications as sensors can be imagined. Model compound 1a efficiently coordinates trivalent metal cations both in water and dimethyl sulfoxide (DMSO). Significantly, the effects of the complexation strongly depend on the aggregation state of the PBI molecules in solution. As a matter of fact, in water, the presence of M(3+) ions triggers the formation of light emitting supramolecular aggregates (excimers). On the other hand, in DMSO-rich solutions metal complexation leads to the suppression of the PET and leads to a strong fluorescence enhancement.


Deformable image registration for the use of magnetic resonance spectroscopy in prostate treatment planning.
There is now convincing evidence that prostate cancer cells lack the ability to produce and accumulate citrate. Using magnetic resonance spectroscopy imaging (MRSI), regions of absent or low citrate concentration in the prostate can be visualized at a resolution of a few mm. This new advancement provides not only a tool for early diagnosis and screening but also the opportunity for preferential targeting of radiation to regions of high tumor burden in the prostate. The differences in the shape and location of the prostate between MRSI imaging and treatment have been the major obstacle in integrating MRSI in radiation therapy treatment planning. The purpose of this study is to develop a reliable method for deforming the prostate and surrounding regions from the geometry of MRSI imaging to the geometry of treatment planning, so that the regions of high tumor burden identified by the MRSI study can be faithfully transferred to the images used for treatment planning. Magnetic resonance spectroscopy imaging studies have been performed on 2 prostate cancer patients using a commercial MRSI system with an endorectal coil and coupling balloon. At the end of each study, we also acquired the MRI of the pelvic region at both the deformed state where the prostate is distorted by the endorectal balloon and the resting state with the endorectal balloon deflated and removed. The task is to find a three-dimensional matrix of transformation vectors for all volume elements that links the two image sets. We have implemented an optimization method to iteratively optimize the transformation vectors using a Newton-Ralphson algorithm. The objective function is based on the mutual information. The distorted images using the transformation vectors are compared with the images acquired at the resting conditions. The algorithm is capable of performing the registration automatically without the need for intervention. It does not require manual contouring of the organs. By applying the algorithm to multiple image sets of different patients, we found a good agreement between the images transformed from those acquired at the deformed state and those acquired at resting conditions. The computation time required for achieving the registration is in the range of a half-hour (for image size: 256 pixels x 256 pixels x 25 slices). However, the space of registration can be restricted to speed up the process. In this article, we described a three-dimensional deformable image registration method to automatically transform images from the deformed imaging state to resting state. Our examples show that this method is feasible and useful to the treatment planning system.


Anatomical distribution and biochemical characterization of the novel RFamide peptide 26RFa in the human hypothalamus and spinal cord.
26RFa is a novel RFamide peptide originally isolated in the amphibian brain. The 26RFa precursor has been subsequently characterized in various mammalian species but, until now, the anatomical distribution and the molecular forms of 26RFa produced in the CNS of mammals, in particular in human, are unknown. In the present study, we have investigated the localization and the biochemical characteristics of 26RFa-like immunoreactivity (LI) in two regions of the human CNS--the hypothalamus and the spinal cord. Immunohistochemical labeling using specific antibodies against human 26RFa and in situ hybridization histochemistry revealed that in the human hypothalamus 26RFa-expressing neurons are located in the paraventricular and ventromedial nuclei. In the spinal cord, 26RFa-expressing neurons were observed in the dorsal and lateral horns. Characterization of 26RFa-related peptides showed that two distinct molecular forms of 26RFa are present in the human hypothalamus and spinal cord, i.e. 26RFa and an N-terminally elongated form of 43 amino acids designated 43RFa. These data provide the first evidence that 26RFa and 43RFa are actually produced in the human CNS. The distribution of 26RF-LI suggests that 26RFa and/or 43RFa may modulate feeding, sexual behavior and transmission of nociceptive stimuli.


The cardiac correlates of feeling safe in everyday life: A Bayesian replication study.
Feeling safe and secure has been proposed to dampen autonomic arousal and buffer threat responses. In a previous study, we could show that momentary ratings of subjective safety were associated with elevated heart rate variability (specifically, root mean square of successive differences; RMSSD) and lower heart rate in everyday life, thus suggesting a health-protective role of feeling safe. This study aimed to replicate this effect in a sample of N = 79 adults, applying Bayesian statistics with prior effects of the original study. Using an ecological momentary assessment (EMA) across three days we could replicate the effect of lower heart rate and higher RMSSD in moments when participants felt more safe. In accordance with the original study, we could also show that the effect on heart rate were independent of RMSSD, thus suggesting a contribution of sympathetic activity to this effect. The findings confirm the connection between momentary feelings of safety and cardiac regulation, thus substantiating research on the health-protective role of psychological safety.


[The application of ultrasonic bone curette in laminoplasty of spinal canal after resection of intraspinal tumors].
To explore the practicability and safety of ultrasonic bone curette in the laminoplasty of spinal canal after resection of intraspinal tumors. The clinical data of 17 patients with thoracolumbar intraspinal tumors treated with ultrasonic bone curette after resection of intraspinal tumors between December 2015 and April 2017 were retrospectively analyzed. All patients were male, aged 42-73 years with an average of 57.4 years. The disease duration was 2-47 months with an average of 21.1 months. Among them, there were 4 cases of thoracic intrathoracic tumors (T 10 in 1, T 12 in 3) and 13 cases of lumbar intrathoracic tumors (L 1 in 5, L 2 in 4, L 3 in 2, and L 4 in 2). Postoperative pathological diagnosis showed that 8 cases were schwannoma, 4 cases were meningioma, 2 cases were neurofibroma, 2 cases were dermoid cyst, and 1 case was ependymoma. Spinal nerve function was evaluated preoperatively according to Frankel classification criteria, with 2 cases of grade B, 7 cases of grade C, and 8 cases of grade D. During the operation, the time of single segmental vertebral canal posterior wall incision, the overall operation time, intraoperative blood loss, intraoperative dural injury, and cerebrospinal fluid leakage, spinal cord and nerve root injury were recorded. At 3-6 months after operation, the tumor and bone healing were observed according to MRI and CT three-dimensional reconstruction, and the spinal nerve function was evaluated by Frankel classification. The time of ultrasonic osteotomy for the posterior wall of a single segmental vertebral canal was 3.4-5.7 minutes, with an average of 4.1 minutes. The overall operation time was 135-182 minutes, with an average of 157.3 minutes. The intraoperative blood loss was 300-500 mL, with an average of 342.6 mL. There was no accidental dural injury, and cerebrospinal fluid leakage, nerve root injury, or spinal cord injury. The incision healed by first intention after operation. All the 17 patients were followed up 9-18 months, with an average of 12.7 months. MRI examination showed no tumor recurrence, and CT three-dimensional reconstruction showed good bone healing in all patients. During the follow-up, there was no loosening or rupture of the internal fixator and there was no re-compression of the spinal cord. At last follow-up, according to Frankel classification, there were 1 case as grade B, 5 cases as grade C, 7 cases as grade D, and 4 cases as grade E. The application of ultrasonic bone curette in laminoplasty of spinal canal after resection of intraspinal tumors can preserve the integrity of the bone ligament structure of posterior column, maintain the volume of vertebral canal, and has high safety, practicability, and good postoperative effectiveness.


Diet intervention improves cardiovascular profile in patients with rheumatoid arthritis: results from the randomized controlled cross-over trial ADIRA.
The chronic inflammation in patients with rheumatoid arthritis (RA) increases the risk for cardiovascular diseases (CVD). The contribution of diet as a risk factor for CVD among these patients is however not fully understood. The aim of this study is to investigate if a proposed anti-inflammatory diet improves cardiovascular profile in weight stable patients with RA. Patients (n = 50) with RA were included in a cross-over trial. They were randomized to either a diet rich in whole grain, fatty fish, nuts, vegetables and fruit and supplemented with probiotics, or a control diet resembling average nutritional intake in Sweden, for ten weeks. After a 4-month washout they switched diet. Participants received food bags and dietary guidelines. Primary outcome was triglyceride (TG) concentration. Secondary outcomes were total-, high density lipoprotein- (HDL) and low density lipoprotein- (LDL) cholesterol, Apolipoprotein-B100 and -A1, lipoprotein composition, plasma phospholipid fatty acids and blood pressure. Forty-seven patients completed at least one period and they remained weight stable. There was a significant between-dietary treatment effect in TG and HDL-cholesterol concentration in favor of intervention (p = 0.007 and p = 0.049, respectively). Likewise, Apolipoprotein-B100/A1 ratio shifted toward a less atherogenic profile in favor of the intervention (p = 0.007). Plasma fatty acids increased in polyunsaturated- and decreased in monounsaturated- and saturated fatty acids between diet periods in favor of the intervention period. Blood lipid profile improved indicating cardioprotective effects from an anti-inflammatory dietary intervention in patients with RA. This trial is registered at ClinicalTrials.gov as NCT02941055 .


HIV-1 pol gene variation is sufficient for reconstruction of transmissions in the era of antiretroviral therapy.
We wished to assess the potential of using HIV-1 pol gene for the identification of transmissions events by phylogenetic means in the era of antiretroviral drug selective pressure. The relatedness of the viruses within a large database of pol sequences generated from HIV-1 infected individuals from the UK was reconstructed by phylogenetic analyses. A total of 140 pol sequences were selected out of the 2500 database entries, on the basis of a pairwise genetic distance higher than 95%. Neighbour Joining and Maximum Likelihood trees were implemented. Trees were reconstructed after exclusion of codon positions associated with drug resistance from the original pol alignment. Trees based on the corresponding env and gag genes were implemented to confirm the linkages. Up to 23 transmission clusters were identified, supported by high bootstrap values (> 99), congruent epidemiological data and/or similar drug resistance motifs. The topology of the tree was consistent after exclusion of the drug resistance associated codons. Identical topologies were obtained in trees implemented from gag and env genes alignments. Despite its genetic conservation, the HIV-1 pol gene holds sufficient variability to permit the phylogenetic reconstruction of transmissions. Identical clusters were obtained whichever of the three principal genes is considered and no bias was induced by the presence of drug resistance mutations. These findings demonstrate the important epidemiological information inherent within routinely collected laboratory data, which can assist in estimating rates of recent HIV-1 transmission within a population.


Functional analysis of monocyte MHC class II compartments.
Circulating monocytes, as dendritic cell and macrophage precursors, exhibit several functions usually associated with antigen-presenting cells, such as phagocytosis and presence of endosomal/lysosomal degradative compartments particularly enriched in Lamp-1, MHC class II molecules, and other proteins related to antigen processing and MHC class II loading [MHC class II compartments (MIICs)]. Ultrastructural analysis of these organelles indicates that, differently from the multivesicular bodies present in dendritic cells, in monocytes the MIICs are characterized by a single perimetral membrane surrounding an electron-dense core. Analysis of their content reveals enrichment in myeloperoxidase, an enzyme classically associated with azurophilic granules in granulocytes and mast cell secretory lysosomes. Elevation in intracellular free calcium levels in monocytes induced secretion of beta-hexosaminidase, cathepsins, and myeloperoxidase in the extracellular milieu; surface up-regulation of MHC class II molecules; and appearance of lysosomal resident proteins. The Ca(2+)-regulated surface transport mechanism of MHC class II molecules observed in monocytes is different from the tubulovesicular organization of the multivesicular bodies previously reported in dendritic cells and macrophages. Hence, in monocytes, MHC class II-enriched organelles combine degradative functions typical of lysosomes and regulated secretion typical of secretory lysosomes. More important, Ca(2+)-mediated up-regulation of surface MHC class II molecules is accompanied by extracellular release of lysosomal resident enzymes.


Psychotic reactivity to daily life stress and the dopamine system: a study combining experience sampling and [18F]fallypride positron emission tomography.
Stressful life events increase the risk for psychosis, and the subjective experience of stress related to daily life activities drives moment-to-moment variation in psychotic intensity. Positron emission tomography (PET) studies suggest that dopaminergic (DAergic) activity mediates the behavioral response to an experimental stressor. However, it is not known how alterations in this DAergic stress response relate to the subjective experience of stress in real life situations assessed in momentary assessment studies. This study combined [18F]fallypride PET with an Experience Sampling ambulatory assessment approach to examine the association between the prefrontal DAergic response to experimentally induced stress and real life psychotic reactivity to the subjective experience of stress in daily life. Healthy first-degree relatives of individuals with a psychotic disorder (N = 14) and healthy controls (N = 11) participated in (a) a psychosocial [18F]fallypride PET stress paradigm and (b) an experience sampling study, using a structured diary approach. Mixed multilevel random intercept models revealed that stress-induced [18F]fallypride displacement, indicative of DAergic activity, in ventromedial prefrontal cortex (VMPFC) was associated with psychotic reactivity to daily life stress in the entire sample. Lower levels of [18F]fallypride displacement to stress predicted increased psychotic reactivity to daily life stress. This study combined PET neuroimaging with real life behavioral assessments in the investigation of psychotic symptoms; we showed decreased [18F]fallypride displacement to stress in VMPFC to be associated with increased psychotic reactivity to daily life stress. The preliminary evidence in this study demonstrates that it is possible to acquire a grasp on how brain function is associated with contextualized experience, which has relevance for neuroimaging studies in general.


Differential conditioning of associative synaptic enhancement in hippocampal brain slices.
An electrophysiological stimulation paradigm similar to one that produces Pavlovian conditioning was applied to synaptic inputs to pyramidal neurons of hippocampal brain slices. Persistent synaptic enhancement was induced in one of two weak synaptic inputs by pairing high-frequency electrical stimulation of the weak input with stimulation of a third, stronger input to the same region. Forward (temporally overlapping) but not backward (temporally separate) pairings caused this enhancement. Thus hippocampal synapses in vitro can undergo the conditional and selective type of associative modification that could provide the substrate for some of the mnemonic functions in which the hippocampus is thought to participate.


Self-rated health and adverse events in CKD.
Little is known about the utility of self-rated general health assessments in persons with moderate-to-severe CKD. This study examined the ability of a single self-rated health measure to predict all-cause mortality and kidney disease progression in a cohort of 443 patients with stages 3-4 CKD, recruited between 2005 and 2011, and followed until the end of 2012. The performance of models incorporating self-rated health measures was compared with previously published predictive models and more complex models comprising a multibiomarker panel. Participants were asked "In general, would you say your health is excellent, very good, good, fair, or poor?" Outcomes examined were time to all-cause mortality, kidney disease progression (initiation of RRT or 30% loss of eGFR), and a composite of these events. Model performances were compared using a nonparametric area under the curve (AUC) analysis. Over a median follow-up of 3.3 years, 118 (27%) participants died and 138 (31%) had progression of kidney disease. Fair-to-poor self-rated health status was associated with significantly greater risks of mortality (fully adjusted hazard ratio [HR] for relative to good-to-excellent self-rated health, 2.76; 95% confidence interval [95% CI], 1.28 to 5.89), kidney disease progression (HR, 1.94; 95% CI, 1.49 to 2.56), and the combined end point (HR, 2.21; 95% CI, 1.66 to 2.96). For 3-year mortality prediction, the self-rated health model (AUC, 0.80; 95% CI, 0.76 to 0.85) had significantly higher AUCs than the base model (AUC, 0.71; 95% CI, 0.66 to 0.76) and the multibiomarker panel model (AUC, 0.74; 95% CI, 0.68 to 0.80) (P=0.03 and P=0.04, respectively). A single, easily obtained measure of self-rated health helps identify patients with CKD at high risk of mortality and kidney disease progression. Routine evaluation of self-rated health may help target individuals who might benefit from more intensive monitoring strategies.


[Generalized calcinosis of the internal organs associated with multiple endocrine adenomatosis].
A rare observation of associated universal calcinosis of the internal organs and multiple adenomatosis is described. The patient was diagnosed to have renal cystadenoma with a high content of neutral lipids in the cells; medullary thyroid carcinoma; nodular adrenal hyperplasia; microadenoma of eosinophilic cells and basophilic cell hyperplasia of the anterior hypophysis; microgangliocytoma of the hypophyseal peduncle; atrophy of the parathyroid gland. Symptoms of hypercalcemia, hypercorticoidism, and some features of the carcinoid syndrome were clinically observed. It is suggested that the clinical symptoms were due to the increased production of vitamin D3 active metabolites by the renal tumour.


Recent advancements in optofluidics-based single-cell analysis: optical on-chip cellular manipulation, treatment, and property detection.
Cellular analysis plays important roles in various biological applications, such as cell biology, drug development, and disease diagnosis. Conventional cellular analysis usually measures the average response from a whole cell group. However, bulk measurements may cause misleading interpretations due to cell heterogeneity. Another problem is that current cellular analysis may not be able to differentiate various subsets of cell populations, each exhibiting a different behavior than the others. Single-cell analysis techniques are developed to analyze cellular properties, conditions, or functional responses in a large cell population at the individual cell level. Integrating optics with microfluidic platforms provides a well-controlled microenvironment to precisely control single cell conditions and perform non-invasive high-throughput analysis. This paper reviews recent developments in optofluidic technologies for various optics-based single-cell analyses, which involve single cell manipulation, treatment, and property detection. Finally, we provide our views on the future development of integrated optics with microfluidics for single-cell analysis and discuss potential challenges and opportunities of this emerging research field in biological applications.


Changing landscape of paediatric refugee health in South Western Sydney, Australia: a retrospective observational study.
To examine the changing health needs of refugee children and young people (CYP) entering Australia, in relation to key government policy changes. Retrospective analysis of health service use data over 11 years. Paediatric refugee clinics in South Western Sydney (SWS), the Australian region with the largest annual resettlement of refugees. Refugee CYP (≤25 years) attending the SWS paediatric refugee clinics for their first visit between 2009 and 2019. Clinician defined health conditions categorised as communicable and non-communicable disease (NCD). Data were analysed for 359 CYP, mean age 9.3 years; 212 male (59.1%). Most CYP (n=331, 92.2%) had health problems identified; 292 (81.3%) had ≥1 NCD and 24 (6.7%) had ≥1 communicable disease. The most frequent individual NCDs were dental disease (n=128, 35.7%) and vitamin D deficiency (n=72, 20.1%). Trend analysis showed increased odds of identifying an NCD from 2013 onwards (crude OR 1.77, 95% CI 1.06 to 2.96). Neurodevelopmental problems, especially Global Developmental Delay (n=31, 8.6%), emerged as more prevalent issues in the latter half of the decade. There were significantly increased odds of identifying a neurodevelopmental problem in 2016-2019, especially in 2016-2017 (adjusted OR 2.93, 95% CI 1.34 to 6.40). Key policy changes during this period included acceptance of refugees with disabilities from 2012, additional Australian Humanitarian Programme intake from the Eastern Mediterranean region and mandatory offshore processing for those seeking asylum by boat from 2013. In response to the changing needs, local health services adopted nurse-led primary healthcare screening, early childhood services, youth and disability clinics. Refugee CYP in Australia are presenting with a growing burden of NCDs, with neurodevelopmental problems contributing. Government policy changes affect the sociodemographics of resettled populations, influencing health profiles. Paediatric refugee health services need to be responsive to the changing needs of these populations to optimise well-being.


Injection of chemotherapeutic microspheres and glioma. III: Parameters to optimize efficacy.
Injectable microspheres may provide a means of providing local, sustained exposure of glioma to chemotherapeutics to improve patient survival. Using a rodent model of surgically resected glioma, we previously demonstrated that direct injections of chemotherapeutic microspheres into the tissue surrounding a resection cavity provide superior survival effects over injections of the same microspheres directly into the surgical cavity. The present experiments extended this novel observation by exploring several parameters related to the use of intraparenchymal injections of chemotherapeutic microspheres to treat glioma. Using a rat model of resected glioma, several principles regarding the use of local sustained release carboplatin microspheres were established. First, an inverted U dose-response was observed, wherein further dose escalation beyond the optimal dose was not efficacious and indeed produced significant local toxicity. Second, it was necessary to expose approximately 40% of the tumor margin to sustained release carboplatin in order to increase survival in this model. Survival was not enhanced when the proportion of the tumor margin exposed to carboplatin was only 20%. Third, the distribution of the chemotherapeutic microsphere injections along the tumor perimeter was shown to be important, requiring that the entire perimeter be proportionately exposed to the chemotherapeutic agent. Together, these data continue to support the development of chemotherapeutic microspheres for treating glioma. However, they also caution that a number of fundamental parameters can profoundly influence the efficacy that might be expected from local sustained delivery. Careful attention to these principles is not only required if chemotherapeutic microspheres are to be used efficaciously, but these principles should provide a foundation to further optimize the potential of this and other polymeric delivery systems under development for local, intraparenchymal drug delivery to glioma.


[Comparative morphological study of thyroid gland in mammalian animals and man].
The objective of this study was to examine the morphological peculiarities of the thyroid gland in animals of different mammalian orders and in man at both macroscopic and microscopic level. 304 specimens of the thyroid gland obtained from the animals of 13 species and 5 mammalian orders and man, were studied. At the macroscopic level, the changes of lobar shape, presence and structural peculiarities of the isthmus, and its relative size were analyzed. At the microscopic level, the following parameters were determined in comparative aspect: the volume fractions of major structural components, the indices of activity, sclerosis and colloid accumulation, the standardized follicular profile shape and size, follicular epithelium height. With the increase of size of the species studied, a tendency was found for the increase of thyroid lobe width and the appearance of parenchymal isthmus, connecting the lobes. At the microscopic level, the variability of follicular profile shape and size was found to increase from small rodents to artiodactyls and man. According to morphological indices, the activity of the thyroid gland in artiodactyls and man was decreased as compared to that one in the other mammalian species.


Neck circumference as a potential indicator of pre-sarcopenic obesity in a cohort of community-based individuals.
ESPEN/EASO advocates screening for sarcopenic obesity based on the concomitant presence of an elevated body mass index (BMI) or waist circumference. Neck circumference (NC) is another simple and reliable anthropometric measurement for estimating obesity; however, its ability to detect sarcopenic obesity has not yet been established. The aim of the present study was to explore the association between NC and sarcopenic obesity in a Shanghai community population. The study included 1542 participants (622 men and 920 women) with a mean age of 58 years who underwent an examination for the detection of obesity at baseline in 2013-2014 and received a re-examination in 2015-2016. An automatic bioelectric impedance analyzer was used to estimate body composition, and magnetic resonance imaging was used to measure abdominal fat distribution. The definition of pre-sarcopenic obesity combined low skeletal muscle mass adjusted by weight (SMM/W) with obesity which defined according to overall adiposity or fat distribution as BMI ≥25 kg/m2, fat percentage (fat%) ≥ 25% in men and 30% in women, or visceral fat area (VFA) ≥ 80 cm2, respectively. In both men and women, subjects with low SMM/W had a higher level of NC than those without (both P < 0.01). In turn, participants with elevated NC had a higher proportion of pre-sarcopenic obesity in both men and women, regardless of adiposity status assessed by BMI, fat%, or VFA (all P < 0.01). During an average follow up of 2.1 years, for each 1 cm increase in NC, multivariable-adjusted hazard ratios of pre-sarcopenic obesity in which adiposity status assessed by high BMI were 1.40 (1.11-1.76) in men and 1.32 (1.13-1.56) in women; in addition, such association remained between NC and pre-sarcopenic obesity assessed by high fat% or high VFA. NC is closely associated with the incidence of sarcopenic obesity, suggesting that it could be helpful for screening sarcopenic obesity in a community-based population.


Recombinant leptin for weight loss in obese and lean adults: a randomized, controlled, dose-escalation trial.
The protein hormone leptin is important to the homeostatic regulation of body weight. Treatment with exogenous leptin may affect weight loss. To determine the relationship between increasing doses of exogenous leptin administration and weight loss in both lean and obese adults. A randomized, double-blind, placebo-controlled, multicenter, escalating dose cohort trial conducted from April 1997 to October 1998. Four university nutrition and obesity clinics and 2 contract clinical research clinics. Fifty-four lean (body mass index, 20.0-27.5 kg/m2; mean [SD] body weight, 72.0 [9.7] kg) and 73 obese (body mass index, 27.6-36.0 kg/m2; mean [SD] body weight, 89.8 [11.4] kg) predominantly white (80%) men (n = 67) and women (n = 60) with mean (SD) age of 39 (10.3) years. Recombinant methionyl human leptin self-administered by daily morning subcutaneous injection (0 [placebo], 0.01, 0.03, 0.10, or 0.30 mg/kg). In part A, lean and obese subjects were treated for 4 weeks; in part B, obese subjects were treated for an additional 20 weeks. Lean subjects consumed a eucaloric diet to maintain body weight at the current value, and obese subjects were prescribed a diet that reduced their daily energy intake by 2100 kJ/d (500-kcal/d) from the amount needed to maintain a stable weight. Body weight, body fat, and incidence of adverse events. Weight loss from baseline increased with increasing dose of leptin among all subjects at 4 weeks (P = .02) and among obese subjects at 24 weeks (P = .01) of treatment. Mean (SD) weight changes at 4 weeks ranged from -0.4 (2.0) kg for placebo (n = 36) to -1.9 kg (1.6) kg for the 0.1 mg/kg dose (n = 29). Mean (SD) weight changes at 24 weeks ranged from -0.7 (5.4) kg for the 0.01 mg/kg dose (n = 6) to -7.1 (8.5) kg for the 0.30 mg/kg dose (n = 8). Fat mass declined from baseline as dose increased among all subjects at 4 weeks (P = .002) and among obese subjects at 24 weeks of treatment (P = .004); more than 95% of weight loss was fat loss in the 2 highest dose cohorts at 24 weeks. Baseline serum leptin concentrations were not related to weight loss at week 4 (P = .88) or at week 24 (P = .76). No clinically significant adverse effects were observed on major organ systems. Mild-to-moderate reactions at the injection site were the most commonly reported adverse effects. A dose-response relationship with weight and fat loss was observed with subcutaneous recombinant leptin injections in both lean and obese subjects. Based on this study, administration of exogenous leptin appears to induce weight loss in some obese subjects with elevated endogenous serum leptin concentrations. Additional research into the potential role for leptin and related hormones in the treatment of human obesity is warranted.


Exercise plasma metabolomics and xenometabolomics in obese, sedentary, insulin-resistant women: impact of a fitness and weight loss intervention.
Insulin resistance has wide-ranging effects on metabolism, but there are knowledge gaps regarding the tissue origins of systemic metabolite patterns and how patterns are altered by fitness and metabolic health. To address these questions, plasma metabolite patterns were determined every 5 min during exercise (30 min, ∼45% of V̇o2peak, ∼63 W) and recovery in overnight-fasted sedentary, obese, insulin-resistant women under controlled conditions of diet and physical activity. We hypothesized that improved fitness and insulin sensitivity following a ∼14-wk training and weight loss intervention would lead to fixed workload plasma metabolomics signatures reflective of metabolic health and muscle metabolism. Pattern analysis over the first 15 min of exercise, regardless of pre- versus postintervention status, highlighted anticipated increases in fatty acid tissue uptake and oxidation (e.g., reduced long-chain fatty acids), diminution of nonoxidative fates of glucose [e.g., lowered sorbitol-pathway metabolites and glycerol-3-galactoside (possible glycerolipid synthesis metabolite)], and enhanced tissue amino acid use (e.g., drops in amino acids; modest increase in urea). A novel observation was that exercise significantly increased several xenometabolites ("non-self" molecules, from microbes or foods), including benzoic acid-salicylic acid-salicylaldehyde, hexadecanol-octadecanol-dodecanol, and chlorogenic acid. In addition, many nonannotated metabolites changed with exercise. Although exercise itself strongly impacted the global metabolome, there were surprisingly few intervention-associated differences despite marked improvements in insulin sensitivity, fitness, and adiposity. These results and previously reported plasma acylcarnitine profiles support the principle that most metabolic changes during submaximal aerobic exercise are closely tethered to absolute ATP turnover rate (workload), regardless of fitness or metabolic health status.


Structure-function analysis of USP1: insights into the role of Ser313 phosphorylation site and the effect of cancer-associated mutations on autocleavage.
In complex with its cofactor UAF1, the USP1 deubiquitinase plays an important role in cellular processes related to cancer, including the response to DNA damage. The USP1/UAF1 complex is emerging as a novel target in cancer therapy, but several aspects of its function and regulation remain to be further clarified. These include the role of the serine 313 phosphorylation site, the relative contribution of different USP1 sequence motifs to UAF1 binding, and the potential effect of cancer-associated mutations on USP1 regulation by autocleavage. We have generated a large set of USP1 structural variants, including a catalytically inactive form (C90S), non-phosphorylatable (S313A) and phosphomimetic (S313D) mutants, deletion mutants lacking potential UAF1 binding sites, a mutant (GG/AA) unable to undergo autocleavage at the well-characterized G670/G671 diglycine motif, and four USP1 mutants identified in tumor samples that cluster around this cleavage site (G667A, L669P, K673T and A676T). Using cell-based assays, we have determined the ability of these mutants to bind UAF1, to reverse DNA damage-induced monoubiquitination of PCNA, and to undergo autocleavage. A non-phosphorylatable S313A mutant of USP1 retained the ability to bind UAF1 and to reverse PCNA ubiquitination in cell-based assays. Regardless of the presence of a phosphomimetic S313D mutation, deletion of USP1 fragment 420-520 disrupted UAF1 binding, as determined using a nuclear relocation assay. The UAF1 binding site in a second UAF1-interacting DUB, USP46, was mapped to a region homologous to USP1(420-520). Regarding USP1 autocleavage, co-expression of the C90S and GG/AA mutants did not result in cleavage, while the cancer-associated mutation L669P was found to reduce cleavage efficiency. USP1 phosphorylation at S313 is not critical for PCNA deubiquitination, neither for binding to UAF1 in a cellular environment. In this context, USP1 amino acid motif 420-520 is necessary and sufficient for UAF1 binding. This motif, and a homologous amino acid segment that mediates USP46 binding to UAF1, map to the Fingers sub-domain of these DUBs. On the other hand, our results support the view that USP1 autocleavage may occur in cis, and can be altered by a cancer-associated mutation.


Expression, purification, crystallization and preliminary crystallographic analysis of spermidine acetyltransferase from Escherichia coli.
The spermidine acetyltransferase (SAT) from Escherichia coli catalyses the transfer of acetyl groups from acetyl-CoA to spermidine. SAT has been expressed and purified from E. coli. SAT was crystallized by the sitting-drop vapour-diffusion method to obtain a more detailed insight into the molecular mechanism. Preliminary X-ray diffraction studies revealed that the crystals diffracted to 2.5 Å resolution and belonged to the cubic space group P23, with unit-cell parameters a = b = c = 148.7 Å. They contained four molecules per asymmetric unit.


Molecular Infectious Disease Epidemiology: Survival Analysis and Algorithms Linking Phylogenies to Transmission Trees.
Recent work has attempted to use whole-genome sequence data from pathogens to reconstruct the transmission trees linking infectors and infectees in outbreaks. However, transmission trees from one outbreak do not generalize to future outbreaks. Reconstruction of transmission trees is most useful to public health if it leads to generalizable scientific insights about disease transmission. In a survival analysis framework, estimation of transmission parameters is based on sums or averages over the possible transmission trees. A phylogeny can increase the precision of these estimates by providing partial information about who infected whom. The leaves of the phylogeny represent sampled pathogens, which have known hosts. The interior nodes represent common ancestors of sampled pathogens, which have unknown hosts. Starting from assumptions about disease biology and epidemiologic study design, we prove that there is a one-to-one correspondence between the possible assignments of interior node hosts and the transmission trees simultaneously consistent with the phylogeny and the epidemiologic data on person, place, and time. We develop algorithms to enumerate these transmission trees and show these can be used to calculate likelihoods that incorporate both epidemiologic data and a phylogeny. A simulation study confirms that this leads to more efficient estimates of hazard ratios for infectiousness and baseline hazards of infectious contact, and we use these methods to analyze data from a foot-and-mouth disease virus outbreak in the United Kingdom in 2001. These results demonstrate the importance of data on individuals who escape infection, which is often overlooked. The combination of survival analysis and algorithms linking phylogenies to transmission trees is a rigorous but flexible statistical foundation for molecular infectious disease epidemiology.


An experimental model for calcium carbonate urolithiasis in goats.
Calcium carbonate is a common urolith type in small ruminants with no high-yield experimental model to evaluate animal susceptibility or preventative measure response. That novel plastic winged implants would allow accumulation and quantification of calcium carbonate calculus formation in goats on a high-calcium diet and identify individual variation between goats in the mass of calculi produced. Eight nonpregnant 3- and 4-year-old Boer-cross does, weighing 22.3-39.5 kg, determined to be healthy based on physical examination, were used in these experiments. Prospective cohort study for in vivo experimental model development. Implants were placed into the urinary bladder lumen in 8 goats over 2 evaluation periods. The alfalfa-based ration had a total ration Ca : P of 3.29 and 3.84 : 1, respectively. Urine was collected at 0, 28, 56, and 84 days in the 1st experiment; blood and urine at those timepoints in the 2nd experiment. For each evaluation period, the implants were removed 84 days after implantation and weighed. Accumulated calculi mass was calculated and compared between goats and was analyzed for composition. Implant retention was 100% and 86% in the 2 studies. All goats with retained implants accumulated calcium carbonate at a mean implant gain per day across studies ranging from 0.44 to 57.45 mg. Two goats accumulated (0.44-7.65 mg/day and 33.64 & 57.45 mg/day) significantly more urolith material than the cohort across both studies (P = .047). No routine analytes on blood or urine were found to be explanatory for the difference observed. These findings form a basis for implant and diet selection for use in future studies of urolithiasis development and for studies regarding individual susceptibility to urolithiasis.


Surgical Outcome of Reduction and Instrumented Fusion in Lumbar Degenerative Spondylolisthesis.
Lumbar degenerative spondylolisthesis (LDS) is a degenerative slippage of the lumbar vertebrae. We aimed to evaluate the surgical outcome of degenerative spondylolisthesis with neural decompression, pedicular screw fixation, reduction, and posterolateral fusion. This before-after study was carried out on 45 patients (37 female and 8 male) with LDS operated from August 2008 to January 2011. The patients' pain and disability were assessed by visual analogue scale (VAS) and Oswestry disability index (ODI) questionnaire. In surgery, we applied distraction force to facilitate slip reduction. All the intra- and postoperative complications were recorded. The paired t-test and Pearson correlation coefficient were used for statistical analysis. The mean age of patients and mean follow-up period were 58.3±3.5 years and 31.2±4.8 months, respectively. The mean slip correction rate was 52.2% with a mean correction loss of 4.8%. Preoperative VAS and ODI improved from 8.8 and 71.6 to postoperative 2.1 and 28.7, respectively. Clinical improvement was more prominent in more reduced patients, but Pearson coefficient could not find a significant correlation. Although spinal decompression with fusion and posterior instrumentation in surgical treatment of the patients with LDS result in satisfactory outcome, vertebral reduction cannot significantly enhance the clinical improvement.


[Simulation of the quality of life in patients with chronic obstructive pulmonary disease].
The statistically significant formulas calculated on the basis of statistical processing of the data of the Saint George test and physical, functional, laboratory, and radiation studies in 117 patients with chronic obstructive pulmonary disease (COPD) are given. Based on the external function test, the study has provided highly significant statistical formulas for calculating the subjective estimate of the severity of COPD, the degree of its associated psychological problems and overall distress due to COPD in the presence of moist rales and chronic cor pulmonale or in its absence.


Self-injurious behaviour and body site preference.
Self-injury by people with intellectual disabilities is a highly problematic and damaging behaviour with profound implications for quality of life. To date, very little detailed descriptive information has been available on the distribution and location of body sites that are injured. This study presents preliminary information on the locations of the self-injury body sites of 29 school-age individuals with developmental and intellectual disabilities who self-injured daily. Teaching staff in school-based special education programmes independently recorded the body locations of where their student's self-injured. Approximately 80% of the reported self-injury was directed disproportionately toward the head and hands. Three-quarters of head-directed self-injury was located on the front of the head, and 83% of hand-directed self-injury was located on the back of the hands. Furthermore, 32% of the body sites toward which self-injury was directed were located on stimulation-produced analgesia body sites. The implications of these findings are discussed in relation to the opioid hypothesis of self-injurious behaviour, and with regard to future research investigating functional diagnostic strategies considering both social and biological variables.


Anticancer effect of superoxide dismutase on canine mammary gland tumour in vitro.
Reactive oxygen species (ROS) have been shown to promote tumour growth and metastasis in human cell lines. The superoxide anion (•O2 - ) is produced during ROS formation and is involved in tumour cell signalling. Superoxide dismutase (SOD) has been applied to canine mammary gland tumours to investigate its antitumour effects in vitro. Cell proliferation, cell cycle cell migration assays, reverse transcription-quantitative polymerase chain reaction, and western blot analysis were performed to determine the effects of SOD on canine mammary tumour cell line. SOD treatment resulted in anti-proliferative effects and mediated cell cycle arrest in the canine mammary gland tumour cell lines (CIPp and CIPm). It also downregulated the expression of N-cadherin and Vimentin. The results confirmed that SOD inhibits tumour cell proliferation and migration, thus supporting the potential applications of SOD as a chemotherapeutic agent for canine mammary gland tumours.


The role of high-mobility group box-1 in renal ischemia and reperfusion injury and the effect of ethyl pyruvate.
High mobility group box-1(HMGB1) was identified as a DNA-binding protein that functions as a cofactor for proper transcriptional regulation in somatic cells. Extracellular HMGB1 acts as a potent proinflammatory cytokine that contributes to the pathogenesis of diverse inflammatory and infectious disorders. Ethyl pyruvate (EP), a stable aliphatic ester derived from pyruvic acid, was first described as a pharmacological inhibitor of HMGB1 secretion. We designed this study to identify changes in HMGB1 expression in rat kidney tissues after ischemia reperfusion injury and effects of EP on the expression of HMGB1. Sprague-Dawley rats (200-300 g) were subjected to 40 minutes of renal warm ischemia. The animals were divided into 3 groups: sham group without warm ischemia, EP group (EP given before ischemia), and ischemic control group. Kidneys were harvested and serum creatinine and TNF-alpha measured at 6 hours, 1 day, 3 days, and 5 days after reperfusion. We performed immunohistochemical staining of HMGB1. Serum creatinine and TNF-alpha level were elevated in the ischemic control group and the EP injection group. In the EP injection group, serum creatinine and TNF-alpha levels were lower than the ischemic control group. In the 40-minute ischemia-reperfusion model, HMGB1 expression increased at 6 hours after reperfusion and decreased gradually at 1, 3, and 5 days after reperfusion. HMGB1 expression was more distinct at the outer medullary area. intraperitoneal EP injection had no effect on HMGB1 expression. From these results, we deduced that the preventive effect of EP on rat kidney ischemia-reperfusion injury was not due to the decreased expression of HMGB1 but the prevention of HMGB1 release.


Epidemiologic studies of eosinophilic meningitis in southern Taiwan.
A study of the epidemiologic characteristics of 125 cases of eosinophilic meningitis or meningoencephalitis, probably caused by Angiostrongylus cantonensis, which occurred in southern Taiwan in 1968 and 1969 revealed a close association of the disease with the rainy season. In contrast to findings in other geographic areas, most such cases in this study occurred among children. A higher attack rate was observed among aborigines than among descendants of mainland Chinese. Most patients had eaten the giant African snail, Achatina fulica, prior to their illness and this mollusc was commonly found infected with third-stage larvae of A. cantonensis. However, in almost all instances the snail was eaten after it had been thoroughly cooked and examination of cooked snail meat revealed only dead larvae. It is suspected that patients became infected by inadvertently ingesting A. cantonensis larvae liberated when the snails were prepared for consumption.


Potential Impact of Sex and BMI on Response to Therapy in Psoriatic Arthritis: Post Hoc Analysis of Results From the SEAM-PsA Trial.
In this post hoc analysis, we examined the potential impact of sex and BMI on response in the Study of Etanercept and Methotrexate in Combination or as Monotherapy in Subjects with Psoriatic Arthritis (SEAM-PsA) trial (NCT02376790), a 48-week, phase III, randomized controlled trial that compared outcomes with methotrexate (MTX) monotherapy, etanercept (ETN) monotherapy, and MTX+ETN combination therapy in patients with psoriatic arthritis (PsA) who were naïve to MTX and biologics. We evaluated key outcomes at week 24 stratified by sex (male vs female) and BMI (kg/m2; ≤ 30 vs > 30), including the American College of Rheumatology 20 (ACR20) criteria, minimal disease activity (MDA), very low disease activity (VLDA), and Psoriatic Arthritis Disease Activity Score (PASDAS). We analyzed data using descriptive statistics, normal approximation, logistic model, and analysis of covariance. A total of 851 patients completed the SEAM-PsA trial. Higher proportions of men than women who received MTX+ETN combination therapy achieved ACR20 (71.5% vs 58.3%; P = 0.02), MDA (45.8% vs 25.2%; P = 0.0003), and VLDA (19.1% vs 9.5%; P = 0.03), and men achieved better PASDAS (-3.0 vs -2.3; P = 0.0004). Patients with BMI ≤ 30 generally had better outcomes than those with BMI > 30 in some treatment arms for ACR20, MDA, VLDA, and PASDAS; however, there was no consistent pattern regarding the treatment arm in which the difference occurred. Improved outcomes were observed more in men than in women for MDA and PASDAS with MTX+ETN combination therapy. Patients with BMI ≤ 30 had better outcomes than those with BMI > 30, with no clear pattern regarding treatment received. These findings suggest that contextual factors such as sex and BMI may affect response to PsA therapy.


Suicide rates in the world: 1950-2004.
The cross-country differences and the trends of suicide rates in 71 countries from 1950 to 2004 are described. The data are from the World Health Organization's Mortality Database. It shows that suicide rates vary greatly across countries, even within the same region or at similar levels of development. Random-effect models were used to examine the between-country and within-country stabilities in suicide rates. The results show that more than 90% of the variance in suicide rates is due to between-country differences, suggesting suicide rates display a strong temporal stability.


Coronary tone modulation: formation and actions of prostaglandins, endoperoxides, and thromboxanes.
Exogenous prostaglandin (PGE2) contracts bovine and human coronary arteries but its precursor, arachidonic acid, relaxes them. The endoperoxides PGH2 and PGH3 relax bovine coronary strips, but PGH1 produces contraction. The primary prostaglandins exert opposite effects to their own endoperoxide precursors, thus, PGE2 and PGE3 contract, and PGE1 relaxes the bovine coronary arteries. The paradoxical coronary dilation produced by the arachidonate or the PGH2 suggest that little if any coronary isomerase which converts endoperoxide into PGE2 exists, or that a novel, potent, PG-like substance is produced by the isolated coronary arteries. Although the coronaries do not possess thromboxane A2 synthetase activity, the vessels are profoundly contracted by exogenous thromboxane A2. Thromboxane A2 can be synthesized and released by circulating platelets when they are aggregated by endothelial injury or thrombin. Thus, coronary tone, and possible spasm, in ischemic myocardial zones may be influenced markedly by interplay between prostaglandins, endoperoxides, and thromboxane formed by platelets on the one hand, and endoperoxide products synthesized endogenously in the coronary arteries on the other.


Effect of prophylactic amiodarone on mortality after acute myocardial infarction and in congestive heart failure: meta-analysis of individual data from 6500 patients in randomised trials. Amiodarone Trials Meta-Analysis Investigators.
There have been 13 randomised controlled trials of prophylactic amiodarone in patients with recent myocardial infarction (MI) or congestive heart failure (CHF). None of these was powered to detect a mortality reduction of about 20%. We undertook a meta-analysis, based on data from individual patients, to provide a more sensitive and accurate assessment of the benefits and risks of prophylactic amiodarone. Individual data from the studies were abstracted according to a predefined protocol. The summary odds ratios were calculated according to standard methods. There were eight post-MI and five CHF trials; nine trials were double-blind and placebo-controlled, and four compared amiodarone with usual care. 6553 patients were randomly assigned treatment, of which 78% were in post-MI trials and 22% in CHF trials. 89% had had previous MI. The mean left-ventricular ejection fraction was 31%, and median frequency of ventricular premature depolarisation 18 per h. Total mortality was reduced by 13% (odds ratio 0.87 [95% CI 0.78-0.99], p = 0.030) based on classic fixed-effects meta-analysis and by 15% (0.85 [0.71-1.02], p = 0.081) with the more conservative random-effects approach. Arrhythmic/sudden death was reduced by 29% (0.71 [0.59-0.85], p = 0.0003). There was no effect on non-arrhythmic deaths (1.02 [0.87-1.19], p = 0.84). There was no difference in treatment effect between post-MI and CHF studies. The risk of arrhythmic/sudden death in control-group patients was higher in CHF than in post-MI studies (10.7 vs 4.1%), and the best single predictor of risk of arrhythmic/sudden death among all patients was symptomatic CHF. The excess (amiodarone minus control) risk of pulmonary toxicity was 1% per year. Prophylactic amiodarone reduces the rate of arrhythmic/sudden death in high-risk patients with recent MI or CHF and this effect results in an overall reduction of 13% in total mortality.


LabPatch, an acquisition and analysis program for patch-clamp electrophysiology.
An acquisition and analysis program, "LabPatch," has been developed for use in patch-clamp research. LabPatch controls any patch-clamp amplifier, acquires and records data, runs voltage protocols, plots and analyzes data, and connects to spreadsheet and database programs. Controls within LabPatch are grouped by function on one screen, much like an oscilloscope front panel. The software is mouse driven, so that the user need only point and click. Finally, the ability to copy data to other programs running in Windows 95/98, and the ability to keep track of experiments using a database, make LabPatch extremely versatile. The system requirements include Windows 95/98, at least a 100-MHz processor and 16 MB RAM, a data acquisition card, digital-to-analog converter, and a patch-clamp amplifier. LabPatch is available free of charge at http://www.fhs.mcmaster.ca/huizinga/.


Use of personal protective equipment and operating room behaviors in four surgical subspecialties: personal protective equipment and behaviors in surgery.
To evaluate Universal Precautions (UP) compliance in the operating room (OR). Prospective observational cohort. Trained observers recorded information about (1) personal protective equipment used by OR staff; (2) eyewear, glove, or gown breaks; (3) the nature of sharps transfers; (4) risk-taking behaviors of the OR staff; and (5) needlestick injuries and other blood and body-fluid exposures. Barnes-Jewish Hospital, a 1,000-bed, tertiary-care hospital affiliated with Washington University School of Medicine, St Louis, Missouri. OR personnel in four surgical specialties (gynecologic, orthopedic, cardiothoracic, and general). Procedures eligible for the study were selected randomly. Hand surgery and procedures requiring no or a very small incision (eg, arthroscopy, laparoscopy) were excluded. A total of 597 healthcare workers' procedures were observed in 76 surgical cases (200 hours). Of the 597 healthcare workers, 32% wore regular glasses, and 24% used no eye protection. Scrub nurses and medical students were more likely than other healthcare workers to wear goggles. Only 28% of healthcare workers double gloved, with orthopedic surgery personnel being the most compliant. Sharps passages were not announced in 91% of the surgical procedures. In 65 cases (86%), sharps were adjusted manually. Three percutaneous and 14 cutaneous exposures occurred, for a total exposure rate of 22%. OR personnel had poor compliance with UP. Although there was significant variation in use of personal protective equipment between groups, the total exposure rate was high (22%), indicating the need for further training and reinforcement of UP to reduce occupational exposures.


Prevalence of Patent Foramen Ovale in the Japanese Population- Autopsy Study.
Patent foramen ovale (PFO) can cause ischemic stroke because of paradoxical embolism. Autopsy studies have shown that the prevalence of PFO is 25% in whites or blacks. However, there is a paucity of data on the prevalence of PFO in Asians. The aim of this study was to clarify the prevalence of PFO in the Japanese population. We reviewed 52,717 autopsy reports, which were collected and edited by the Japanese Society of Pathology from 2009 to 2012. Next, we inspected consecutive 103 formalin-fixed specimens that had already been examined by certified pathologists from 2009 to 2013 to find PFO and atrial septal aneurysm (ASA). ASA was defined as ≥10 mm protrusion of the septum into the left or the right atrium. In the database of the Japanese Society of Pathology, the incidence of PFO was 0.08% (43/52,717). Inspection of heart specimens disclosed that the prevalence of PFO was 13.6% (14/103). None of the PFO cases was reported at the original autopsy. PFO was more frequently found in the subjects with ASA (50%) than in those without ASA (9.7%) (P=0.004). PFO is under-reported in autopsy reports. Re-evaluation of heart specimens disclosed that the prevalence of PFO was 13.6%. The prevalence was lower than reported in the past.


[Variation of CAG repeats in coding region of ATXN2 gene in different ethnic groups].
Toinvestigate CAG repeats variation of ATXN2 gene coding region in six ethnic groups that live in comparatively different environments, to evaluate whether these variations are under positive selection, and to find factors driving selection effects, 291 unrelated healthy individuals were collected from six ethnic groups and their STR geneotyping was performed. The frequencies of alleles and genotypes were counted and thereby Slatkin's linearized Fst values were calculated. The UPGMA tree against this gene was constructed. The MDS analysis among these groups was carried out as well. The results from the linearized Fst values indicated that there were significant evolutionary differences of the STR in ATXN2 gene between Hui and Yi groups, but not among the other 4 groups. Further analysis was performed by combining our data with published data obtained from other groups. These results indicated that there were significant differences between Japanese and other groups including Hui, Hani, Yunnan Mongolian, and Inner Mongolian. Both Hui and Mongolian from Inner Mongolia were significantly different from Han. In conclusion, the six ethnic groups had their own distribution characterizations of allelic frequencies of ATXN2 STR, and the potential cause of frequency changes in rare alleles could be the consequence of positive selection.


Phosphonylmethoxyalkyl purine and pyrimidine derivatives for treatment of opportunistic cytomegalovirus and herpes simplex virus infections in murine AIDS.
Murine acquired immunodeficiency syndrome (MAIDS) was induced in C57BL/6 mice following infection with the LP-BM5 retrovirus complex. Infected mice developed splenomegaly, lymphadenopathy and loss of B- and T-cell functions 100 days after virus inoculation. Mice with AIDS were highly susceptible to opportunistic murine cytomegalovirus (MCMV) and herpes simplex virus (HSV-1) infections. The therapeutic activities of two phosphonylmethoxyalkyl derivatives, 9-(2-phosphonylmethoxyethyl)adenine (PMEA) and (S)-1-(3-hydroxy-2-phosphonylmethoxy-propyl)cytosine (HPMPC), were evaluated in MAIDS immunosuppressed mice infected with MCMV or HSV-1. MCMV infection resulted in extensive viral replication in lung, liver and spleen and death occurred five to twelve days post-infection. Treatment with either HPMPC or ganciclovir (DHPG) reduced mortality and viral replication in target organs; however, HPMPC was as effective as DHPG at one-fifth the DHPG dose. Moreover, when a single dose (100 mg/kg) of HPMPC was administered 24 h prior to MCMV infection, it suppressed virus replication at seven and 14 days post-infection, thus resulting in a significant prolongation of life. PMEA was effective against opportunistic HSV-1 infections, but appeared to be less effective than HPMPC against MCMV infections. These results indicate that MAIDS can be used as a model for evaluating antivirals in an immunocompromised host, and suggest that both PMEA and HPMPC may be useful in the treatment of opportunistic CMV and HSV-1 infections.


Ablation of Matrix Metalloproteinase-9 Prevents Cardiomyocytes Contractile Dysfunction in Diabetics.
Elevated expression and activity of matrix metalloproteinase-9 (MMP9) and decreased contractility of cardiomyocytes are documented in diabetic hearts. However, it is unclear whether MMP is involved in the regulation of contractility of cardiomyocytes in diabetic hearts. In the present study, we tested the hypothesis that MMP9 regulates contractility of cardiomyocytes in diabetic hearts, and ablation of MMP9 prevents impaired contractility of cardiomyocytes in diabetic hearts. To determine the specific role of MMP9 in cardiomyocyte contractility, we used 12-14 week male WT (normoglycemic sibling of Akita), Akita, and Ins(2+∕-)/MMP9(-∕-) (DKO) mice. DKO mice were generated by cross-breeding male Ins2(+∕-) Akita (T1D) with female MMP9 knockout (MMP9(-∕-)) mice. We isolated cardiomyocytes from the heart of the above three groups of mice and measured their contractility and calcium transients. Moreover, we determined mRNA and protein levels of sarco-endoplasmic reticulum calcium ATPase-2a (SERCA-2a), which is involved in calcium handling during contractility of cardiomyocytes in WT, Akita, and DKO hearts using QPCR, Western blotting and immunoprecipitation, respectively. Our results revealed that in Akita hearts where increased expression and activity of MMP9 is reported, the rates of shortening and re-lengthening (±dL/dt) of cardiomyocytes were decreased, time to 90% peak height and baseline during contractility was increased, rate of calcium decay was increased, and calcium transient was decreased as compared to WT cardiomyocytes. However, these changes in Akita were blunted in DKO cardiomyocytes. The molecular analyses of SERCA-2a in the hearts showed that it was downregulated in Akita as compared to WT but was comparatively upregulated in DKO. These results suggest that abrogation of MMP9 gene prevents contractility of cardiomyocytes, possibly by increasing SERCA-2a and calcium transients. We conclude that MMP9 plays a crucial role in the regulation of contractility of cardiomyocytes in diabetic hearts.


Association of HMGCR polymorphism with late-onset Alzheimer's disease in Han Chinese.
The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) acts as a potential genetic modifier for Alzheimer's disease (AD). Previous reports identified that HMGCR rs3846662 polymorphism is associated with biosynthesis of cholesterol in AD pathology. In order to assess the involvement of the HMGCR polymorphism in the risk of late-onset AD (LOAD) in northern Han Chinese, we performed a case-control study of 2334 unrelated subjects (984 cases and 1350 age- and gender-matched controls) to evaluate the genotype and allele distributions of the HMGCR rs3846662 with LOAD. The genotype distribution (GG, AG, AA) of rs3846662 was significantly different between LOAD patients and controls (P = 0.003), but the allele distribution did not reach a significant difference (P = 0.614). After adjusting for age, gender and the APOE ε4 status, the minor A allele of rs3846662 was validated as a protective factor for LOAD in dominant model (OR = 0.796, P = 0.02, 95% CI = 0.657-0.965). Interestingly, we observed rs3846662 polymorphism was only significantly associated with LOAD in APOE ε4 non-carriers (OR = 0.735, P = 0.005, 95% CI = [0.593, 0.912]). In conclusion, our study demonstrates A allele of HMGCR rs3846662 acts as a protective factor for LOAD in northern Han Chinese.


High-speed high-resolution imaging of intercellular immune synapses using optical tweezers.
Imaging in any plane other than horizontal in a microscope typically requires a reconstruction from multiple optical slices that significantly decreases the spatial and temporal resolution that can be achieved. This can limit the precision with which molecular events can be detected, for example, at intercellular contacts. This has been a major issue for the imaging of immune synapses between live cells, which has generally required the reconstruction of en face intercellular synapses, yielding spatial resolution significantly above the diffraction limit and updating at only a few frames per minute. Strategies to address this issue have usually involved using artificial activating substrates such as antibody-coated slides or supported planar lipid bilayers, but synapses with these surrogate stimuli may not wholly resemble immune synapses between two cells. Here, we combine optical tweezers and confocal microscopy to realize generally applicable, high-speed, high-resolution imaging of almost any arbitrary plane of interest. Applied to imaging immune synapses in live-cell conjugates, this has enabled the characterization of complex behavior of highly dynamic clusters of T cell receptors at the T cell/antigen-presenting cell intercellular immune synapse and revealed the presence of numerous, highly dynamic long receptor-rich filopodial structures within inhibitory Natural Killer cell immune synapses.


PSD95 regulates morphological development of adult-born granule neurons in the mouse hippocampus.
In the adult hippocampus new neurons are generated in the dentate gyrus from neural progenitor cells. Adult-born neurons integrate into the hippocampal circuitry and contribute to hippocampal function. PSD95 is a major postsynaptic scaffold protein that is crucial for morphological maturation and synaptic development of hippocampal neurons. Here we study the function of PSD95 in adult hippocampal neurogenesis by downregulating PSD95 expression in newborn cells using retroviral-mediated RNA interference. Retroviruses coding for a control shRNA or an shRNA targeting PSD95 (shPSD95) were stereotaxically injected into the dorsal dentate gyrus of 2-month-old C57BL/6 mice. PSD95 knockdown did not affect neuronal differentiation of newborn cells into neurons, or migration of newborn neurons into the granule cell layer. Morphological analysis revealed that newborn neurons expressing shPSD95 showed increased dendritic length and increased number of high-order dendrites. Concomitantly, dendrites from shPSD95-expressing newborn granule neurons showed a reduction in the density of dendritic spines. These results suggest that PSD95 is required for proper dendritic and spine maturation of adult-born neurons, but not for early stages of neurogenesis in the hippocampus.


Vector expression of adenovirus type 5 E1a proteins: evidence for E1a autoregulation.
We transiently expressed adenovirus type C E1a proteins in wild-type or mutant form from plasmid vectors which have different combinations of E1a and simian virus 40 enhancer elements and which contain the DNA replication origin of SV40 and can replicate in COS 7 cells. We measured the levels of E1a mRNA encoded by the vectors and the transition regulation properties of the protein products. Three vectors encoded equivalent levels of E1a mRNA in COS 7 cells: (i) a plasmid encoding the wt 289-amino acid E1a protein (this complemented the E1a deletion mutant dl312 for early region E2a expression under both replicative and nonreplicative conditions); (ii) a vector for the wt 243-amino acid E1a protein (this complemented dl312 weakly and only under conditions of high multiplicities of dl312); (iii) a mutant, pSVXL105, in which amino acid residues-38 through 44 of the 289-amino acid E1a protein (which includes two highly conserved residues) are replaced by 3 novel amino acids (this also complemented dl312 efficiently). A fourth vector, mutant pSVXL3 with which linker substitution shifts the reading frame to encode a truncated 70-amino acid fragment from the amino terminus of the 289-amino acid protein, was unable to complement dl312. Surprisingly, pSVXL3 overexpressed E1a mRNA approximately 30-fold in COS 7 cells in comparison with the other vectors. The pSVXL3 overexpression could be reversed by cotransfection with a wt E1a vector. We suggest that wt E1a proteins regulate the levels of their own mRNAs through the recently described transcription repression functions of the 289- and 243-amino acid E1a protein products and that pSVXL3 fails to autoregulate negatively.


Allied health professionals' views on important outcomes of children's elective lower limb orthopaedic surgery: a qualitative interview study to inform a core outcome set.
A common methodological limitation of research that guides surgical procedure selection for children's elective lower limb orthopaedic surgeries is inconsistent outcome selection. Improving outcome consistency can be achieved through the development of a core outcome set (COS). The aim of this study is to identify which outcomes are considered important for children's elective lower limb orthopaedic surgeries by allied health professionals (AHPs) and explore why they select these outcomes, to inform a COS development project. Online semi-structured interviews were conducted with relevant AHPs. Participants were selected using maximum variation purposive sampling; selection was based on profession and inpatient/outpatient role. The data set was analysed using an inductive and deductive approach to thematic analysis. Four physiotherapists, three orthotists, three prosthetists, and two occupational therapists were interviewed. Most identified outcomes of importance related to "activities and participation". From the data, we conceptualised that AHPs with effective multidisciplinary communication focused on child-centred outcomes, while clinicians with limited multidisciplinary teamwork focused on role-based outcomes. There is concurrence between outcomes identified as important in this study, and other qualitative studies in similar populations. These important outcomes were seldom measured in previous studies or in routine clinical practice.Implications for rehabilitationAllied health professionals (AHPs) prioritise activity and participation outcomes after children's elective lower limb orthopaedic surgery.It is important to the rehabilitation of children after elective lower limb orthopaedic surgery that all involved AHPs collaborate with the wider multidisciplinary team.Multidisciplinary team communication encourages collaborative outcome identification, and discourages role defined outcome focus.


Genetic regulation of the qa gene cluster of Neurospora crassa: induction of qa messenger ribonucleic acid and dependency on qa-1 function.
An in vitro protein-synthesizing system (rabbit reticulocyte) was programmed with total polyadenylated messenger ribonucleic acid from wild type and various mutants in the qa gene cluster of Neurospora crassa. The products of two of the qa genes, quinate dehydrogenase (qa-3+) and dehydroshikimate dehydratase (qa-4+), were identified by specific immunoprecipitation and sodium dodecyl sulfate-slab gel electrophoresis. The results indicated that for both genes induction of a specific enzyme activity by quinic acid depends on the de novo synthesis of a specific polypeptide and on the de novo appearance of specific messenger ribonucleic acid detectable by the in vitro translation assay. Furthermore, the results indicated that the appearance of this messenger ribonucleic acid is under the control of the qa-1 gene. The simplest interpretation of these results appears to be that induction of enzyme activity in the qa system is mediated by events at the transcriptional level.


Treatments for spasticity and pain in multiple sclerosis: a systematic review.
To identify the drug treatments currently available for the management of spasticity and pain in multiple sclerosis (MS), and to evaluate their clinical and cost-effectiveness. Electronic bibliographic databases, National Research Register, MRC Clinical Trials Register and the US National Institutes of Health Clinical Trials Register. Systematic searches identified 15 interventions for the treatment of spasticity and 15 interventions for treatment of pain. The quality and outcomes of the studies were evaluated. Reviews of the treatment of spasticity and pain when due to other aetiologies were also sought. There is limited evidence of the effectiveness of four oral drugs for spasticity: baclofen, dantrolene, diazepam and tizanidine. Tizanidine appears to be no more effective than comparator drugs such as baclofen and has a slightly different side-effects profile. Despite claims that it causes less muscle weakness, there was very little evidence that tizanidine performed any better in this respect than other drugs, although it is more expensive. The findings of this review are consistent with reviews of the same treatments for spasticity derived from other aetiologies. There is good evidence that both botulinum toxin (BT) and intrathecal baclofen are effective in reducing spasticity, and both are associated with functional benefit. However, they are invasive, and substantially more expensive. None of the studies included in the review of pain were designed specifically to evaluate the alleviation of pain in patients with MS and there was no consistency regarding the use of validated outcome measures. It was suggested that, although expensive, the use of intrathecal baclofen may be associated with significant savings in hospitalisation costs in relation to bed-bound patients who are at risk of developing pressure sores, thus enhancing its cost-effectiveness. No studies of cost-effectiveness were identified in the review of pain. There is evidence, albeit limited, of the clinical effectiveness of baclofen, dantrolene, diazepam, tizanidine, intrathecal baclofen and BT and of the potential cost-effectiveness of intrathecal baclofen in the treatment of spasticity in MS. Many of the interventions identified are not licensed for the alleviation of pain or spasticity in MS and the lack of evidence relating to their effectiveness may also limit their widespread use. Indeed, forthcoming information relating to the use of cannabinoids in MS may result in there being better evidence of the effectiveness of new treatments than of any of the currently used drugs. It may therefore be of value to carry out double-blind randomised controlled trials of interventions used in current practice, where outcomes could include functional benefit and impact on quality of life. Further research into the development and validation of outcomes measures for pain and spasticity may also be useful, as perhaps would cost-utility studies.


Oxidants, antioxidants and cardiovascular disease.
Basic and clinical studies in the past decade suggest an involvement of oxygen-derived free radicals in some cardiovascular diseases including atherosclerosis and hypertension. In atherogenesis evidence indicates that low-density lipoprotein/cholesterol must be oxidized before it can be taken up by the monocytes/macrophages to form foam cells which contribute to the characteristic fatty streak. Free radicals are considered responsible for this oxidation. Population studies reveal that hypertensive patients generally have a lower intake of ascorbic acid and possibly other antioxidants. Ascorbic acid deficiency may lead to defective vasodilation and increased blood pressure due to destruction of certain endothelium-dependent relaxing factors by free radicals. Further studies in this area appear justified.


Chloride intracellular channel 4 is critical for the epithelial morphogenesis of RPE cells and retinal attachment.
Retinal detachment is a sight-threatening condition. The molecular mechanism underlying the adhesion between the RPE and photoreceptors is poorly understood because the intimate interactions between these two cell types are impossible to model and study in vitro. In this article, we show that chloride intracellular channel 4 (CLIC4) is enriched at apical RPE microvilli, which are interdigitated with the photoreceptor outer segment. We used a novel plasmid-based transfection method to cell-autonomously suppress CLIC4 in RPE in situ. CLIC4 silenced RPE cells exhibited a significant loss of apical microvilli and basal infoldings, reduced retinal adhesion, and epithelial-mesenchymal transition. Ectopically expressing ezrin failed to rescue the morphological changes exerted by CLIC4 silencing. Neural retinas adjacent to the CLIC4-suppressed RPE cells display severe dysplasia. Finally, a high level of aquaporin 1 unexpectedly appeared at the apical surfaces of CLIC4-suppressed RPE cells, together with a concomitant loss of basal surface expression of monocarboxylate transporter MCT3. Our results suggested that CLIC4 plays an important role in RPE-photoreceptor adhesion, perhaps by modulating the activity of cell surface channels/transporters. We propose that these changes may be attributable to subretinal fluid accumulation in our novel retinal detachment animal model.


Characterization of the role of RILP in cell migration.
Rab-interacting lysosomal protein (RILP) is a regulator of late stages of endocytosis. Recent work proved that depletion of RILP promotes migration of breast cancer cells in wound healing assay, whereas its overexpression influences re-arrangements of actin cytoskeleton. Here, we further characterized the role of RILP in cell migration by analyzing several aspects of this process. We showed that RILP is fundamental also for migration of lung cancer cells regulating cell velocity. RILP silencing did not affect Golgi apparatus nor microtubules reorientation during migration. However, both RILP over-expression and expression of its mutated form, RILP-C33, impair cell adhesion and spreading. In conclusion, our results demonstrate that RILP has important regulatory roles in cell motility affecting migration velocity but also in cell adhesion and cell spreading.


Is enough attention given to climate change in health service planning? An Australian perspective.
Within an Australian context, the medium to long-term health impacts of climate change are likely to be wide, varied and amplify many existing disorders and health inequities. How the health system responds to these challenges will be best considered in the context of existing health facilities and services. This paper provides a snapshot of the understanding that Australian health planners have of the potential health impacts of climate change. The first author interviewed (n=16) health service planners from five Australian states and territories using an interpretivist paradigm. All interviews were digitally recorded, key components transcribed and thematically analysed. Results indicate that the majority of participants were aware of climate change but not of its potential health impacts. Despite this, most planners were of the opinion that they would need to plan for the health impacts of climate change on the community. With the best available evidence pointing towards there being significant health impacts as a result of climate change, now is the time to undertake proactive service planning that address market failures within the health system. If considered planning is not undertaken then Australian health system can only deal with climate change in an expensive ad hoc, crisis management manner. Without meeting the challenges of climate change to the health system head on, Australia will remain unprepared for the health impacts of climate change with negative consequences for the health of the Australian population.


Cancer incidence among Nordic airline cabin crew.
Airline cabin crew are occupationally exposed to cosmic radiation and jet lag with potential disruption of circadian rhythms. This study assesses the influence of work-related factors in cancer incidence of cabin crew members. A cohort of 8,507 female and 1,559 male airline cabin attendants from Finland, Iceland, Norway and Sweden was followed for cancer incidence for a mean follow-up time of 23.6 years through the national cancer registries. Standardized incidence ratios (SIRs) were defined as ratios of observed and expected numbers of cases. A case-control study nested in the cohort (excluding Norway) was conducted to assess the relation between the estimated cumulative cosmic radiation dose and cumulative number of flights crossing six time zones (indicator of circadian disruption) and cancer risk. Analysis of breast cancer was adjusted for parity and age at first live birth. Among female cabin crew, a significantly increased incidence was observed for breast cancer [SIR 1.50, 95% confidence interval (95% CI) 1.32-1.69], leukemia (1.89, 95% CI 1.03-3.17) and skin melanoma (1.85, 95% CI 1.41-2.38). Among men, significant excesses in skin melanoma (3.00, 95% CI 1.78-4.74), nonmelanoma skin cancer (2.47, 95% CI 1.18-4.53), Kaposi sarcoma (86.0, 95% CI 41.2-158) and alcohol-related cancers (combined SIR 3.12, 95% CI 1.95-4.72) were found. This large study with complete follow-up and comprehensive cancer incidence data shows an increased incidence of several cancers, but according to the case-control analysis, excesses appear not to be related to the cosmic radiation or circadian disruptions from crossing multiple time zones.