[Effect of rapamycin in inducing naïve murine effector T cell convert to regulatory T cell].
To study the effect of rapamycin in inducing naïve murine effector T cell (Teff) convert to regulatory T cell (Treg) in vitro. The forkhead box protein 3 (FoxP3) negative Teff were isolated and purified from the spleen and lymph node of C57 BL/6 murines aged 6-8 weeks, then Teff were cultured in three groups with mature dendritic cells (mDC), B cells, and plate coated Anti-CD3. In addition, the control wells and the test wells were prepared in each group, rapamycin were not added in the control wells but added in the test wells with concentrations of 1, 10, 50, and 100 nmol/L. Percentages of FoxP3 positive Treg were examined by flow cytometry after 4 days in Anti-CD3 group and after 6 days in the other two groups. As shown by the flow cytometry, the percentages of FoxP3 positive Treg were as follows in three group: in the mDC group, it was 0.01% in the control well and 0.39%, 0.47%, 0.34%, and 0.26% in test wells; in B cell group, it was 0.01% in the control wells and 5.56%, 5.89%, 7.15%, and 4.72% in the test wells; in Anti-CD3 group, it was 0.93% in the control wells and 1.35%, 1.07%, 1.02%, and 1.19% in test wells. No significant difference was found between the test wells and control wells in the mDC group and Anti-CD3 group; however, the percentages of FoxP3 positive Treg was significantly different between the test wells and control wells in the B cell group (P < 0.01). When B cell is acted as the antigen-presenting cell, rapamycin can effectively induce Teff convert to Treg in vitro.


Structural characterization of Dictyostelium discoideum prespore-specific gene D19 and of its product, cell surface glycoprotein PsA.
The Dictyostelium discoideum cell surface antigen PsA is a glycoprotein which first appears in the multicellular stage soon after tip formation and is selectively expressed on prespore cells. The D19 gene encodes an mRNA sequence which is highly enriched in prespore over prestalk cells in the slug stage. We have determined 81 amino acid residues of N-terminal sequence from immunoaffinity-purified PsA protein and shown this sequence to be identical to the predicted sequence of the D19 gene. There are several short repeat elements close to the C terminus, and unequal crossing-over within these is proposed to account for the size polymorphism observed in PsA protein isolated from different D. discoideum strains. The repeats are proline rich and show similarity to the C-terminal region of the D. discoideum cell adhesion molecule, contact sites A. The extreme C terminus, which is also homologous to contact sites A, is characteristic of proteins attached to the plasma membrane via a glycosyl-phosphatidylinositol link. We have marked the PsA gene by insertion of an oligonucleotide encoding an epitope of the human c-myc protein. A construct containing this gene and 990 base pairs of 5'-flanking region directed correct temporal and spatial mRNA accumulation. We found the marked PsA protein, detected with the human c-myc antibody, to be correctly localized on the surface of cells.


Magnesium and cell energetics: At the junction of metabolism of adenylate and non-adenylate nucleotides.
Free magnesium (Mg2+) represents a powerful signal arising from interconversions of adenylates (ATP, ADP and AMP). This is a consequence of the involvement of adenylate kinase (AK) which equilibrates adenylates and uses defined species of Mg-complexed and Mg-free adenylates in both directions of its reaction. However, cells contain also other reversible Mg2+-dependent enzymes that equilibrate non-adenylate nucleotides (uridylates, cytidylates and guanylates), i.e. nucleoside monophosphate kinases (NMPKs) and nucleoside diphosphate kinase (NDPK). Here, we propose that AK activity is tightly coupled to activities of NMPK and NDPK, linking adenylate equilibrium to equilibria of other nucleotides, and with [Mg2+] controlling the ratios of Mg-chelated and Mg-free nucleotides. This coupling establishes main hubs for adenylate-driven equilibration of non-adenylate nucleotides, with [Mg2+] acting as signal arising from all nucleotides rather than adenylates only. Further consequences involve an overall adenylate control of UTP-, GTP- and CTP-dependent pathways and the availability of substrates for RNA and DNA synthesis.


[Changes of the triple flash electroretinogram after 360 degrees retinotomy and macular translocation for subfoveal choroidal neovascularisation in age-related macular degeneration].
The aim of the present study was to evaluate potential electrophysiological changes after 360 degrees retinotomy and macular translocation for subfoveal choroidal neovascularisation in patients with age-related macular degeneration. A consecutive series of 32 patients suffering from subfoveal choroidal neovascularisation secondary to age-related macular degeneration underwent 360 degrees retinotomy and macular translocation. The scotopic triple-flash ERG (TERG) served as the main parameter of the study and was recorded one day prior to the translocation surgery and no earlier than 4 weeks after the silicone oil removal. The TERG amplitudes were significantly reduced after translocation surgery. Depending on the applied flash luminance the mean amplitude reduction of the first b-wave varied between 67 % (0.2 cds/m (2)) and 74 % (0.03 cds/m (2)). The latency of the second b-wave of the preoperative TERG proved to have predictive power with reference to the postoperative visual acuity. A statistically significant correlation between the latency of the second b-wave for a flash intensity of 0.03 and 0.1 cds/m (2) and the postoperative visual acuity was shown (p = 0,016 and p = 0,049). In accordance with previous studies the present study indicates that a significant electrophysiological decrease is caused by surgical procedures associated with macular translocation. In future, due to predictive parameters of the TERG the preoperative selection of patients could be improved.


Validating the content of pediatric outpatient medical records by means of tape-recording doctor-patient encounters.
Information in 51 tape-recorded physician-patient encounters was compared with information written in the patients' medical records. Diagnoses, chief complaints, scheduled appointments, non-drug therapy, and diagnostic studies were uniformly well-recorded. Medication names were well-recorded but dosages were not. Characteristics of care such as levels of function, probable cause of illness, reason for follow-up, and compliance were recorded poorly. Patients were more likely to known about and understand their diagnosis, and names, dosage, and intended function of their medications when this information was written in the record than when it was not. These findings indicate a relationship between the quality of medical records and the effectiveness of care.


Facilitating Neuron-Specific Genetic Manipulations in Drosophila melanogaster Using a Split GAL4 Repressor.
Efforts to map neural circuits have been galvanized by the development of genetic technologies that permit the manipulation of targeted sets of neurons in the brains of freely behaving animals. The success of these efforts relies on the experimenter's ability to target arbitrarily small subsets of neurons for manipulation, but such specificity of targeting cannot routinely be achieved using existing methods. In Drosophila melanogaster, a widely-used technique for refined cell type-specific manipulation is the Split GAL4 system, which augments the targeting specificity of the binary GAL4-UAS (Upstream Activating Sequence) system by making GAL4 transcriptional activity contingent upon two enhancers, rather than one. To permit more refined targeting, we introduce here the "Killer Zipper" (KZip+), a suppressor that makes Split GAL4 targeting contingent upon a third enhancer. KZip+ acts by disrupting both the formation and activity of Split GAL4 heterodimers, and we show how this added layer of control can be used to selectively remove unwanted cells from a Split GAL4 expression pattern or to subtract neurons of interest from a pattern to determine their requirement in generating a given phenotype. To facilitate application of the KZip+ technology, we have developed a versatile set of LexAop-KZip+ fly lines that can be used directly with the large number of LexA driver lines with known expression patterns. KZip+ significantly sharpens the precision of neuronal genetic control available in Drosophila and may be extended to other organisms where Split GAL4-like systems are used.


Calcium in excitation--contraction coupling of frog skeletal muscle.
A principal step in the process leading to muscle contraction is the intracellular release of Ca2+. We have detected and compared some physical and chemical events that reflect Ca2+ release in contracting frog skeletal muscle cells, described the effects of some agents that are believed to alter intracellular Ca2+ release during contraction, and speculated about the role of Ca2+ release in influencing some of the mechanical properties of frog muscle. The specific physical features recorded were changes in striation spacing, myofibrillar orientation, and force development. The chemical feature was the relative change in intracellular [Ca2+] recorded as light emission from cells microinjected with the Ca2+-sensitive protein aequorin. The presence or absence of a correlation among these variables has been used (i) to evaluate the action of some agents thought to change intracellular Ca2+ release in excitation--contraction (E--C) coupling, (ii) to further substantiate the effects of cell length on Ca2+ release, and (iii) to examine some details of models for E--C coupling. The results showed that potentiating agents enhance and prolong intracellular Ca2+ release without changing the rate of Ca2+ removal during E--C coupling. This extra Ca2+ does not produce the same effect on contractions at all lengths. Contractility is inversely related to cell length, and Ca2+-induced activation is normally less than maximum not only at short lengths but also at optimal striation spacings.


False memories of the future: a critique of the applications of probabilistic reasoning to the study of cognitive processes.
The authors argue that the ways in which people-scientists and laymen-use probabilistic reasoning is predicated on a set of often questionable assumptions that are implicit and frequently go untested. They relate to the correspondence between the terms of a theory and the observations used to validate the theory and to the implicit understandings of intention and prior knowledge that arise between the conveyer and the receiver of information. The authors show several ways in which the use of probabilistic reasoning rests on a priori commitments to a partitioning of an outcome space and demonstrate that there are many more assumptions underlying the use of probabilistic reasoning than are usually acknowledged. They unfold these assumptions to show how several different interpretations of the same results in behavioral decision theory and cognitive psychology are equally well supported by "the facts." They then propose a more comprehensive approach to mapping cognitive processes than those currently used, one that is based on the analysis of all of the relevant alternative interpretations presented in the article.


Role of Lys-226 in the catalytic mechanism of Bacillus stearothermophilus serine hydroxymethyltransferase--crystal structure and kinetic studies.
Serine hydroxymethyltransferase (SHMT), a pyridoxal 5'-phosphate (PLP)-dependent enzyme catalyzes the reversible conversion of l-Ser and tetrahydropteroylglutamate (H(4)PteGlu) to Gly and 5,10-methylene tetrahydropteroylglutamate (CH(2)-H(4)PteGlu). Biochemical and structural studies on this enzyme have implicated several residues in the catalytic mechanism, one of them being the active site lysine, which anchors PLP. It has been proposed that this residue is crucial for product expulsion. However, in other PLP-dependent enzymes, the corresponding residue has been implicated in the proton abstraction step of catalysis. In the present investigation, Lys-226 of Bacillus stearothermophilus SHMT (bsSHMT) was mutated to Met and Gln to evaluate the role of this residue in catalysis. The mutant enzymes contained 1 mol of PLP per mol of subunit suggesting that Schiff base formation with lysine is not essential for PLP binding. The 3D structure of the mutant enzymes revealed that PLP was bound at the active site in an orientation different from that of the wild-type enzyme. In the presence of substrate, the PLP ring was in an orientation superimposable with that of the external aldimine complex of wild-type enzyme. However, the mutant enzymes were inactive, and the kinetic analysis of the different steps of catalysis revealed that there was a drastic reduction in the rate of formation of the quinonoid intermediate. Analysis of these results along with the crystal structures suggested that K-226 is responsible for flipping of PLP from one orientation to another which is crucial for H(4)PteGlu-dependent Calpha-Cbeta bond cleavage of l-Ser.


Melatonin, bakuchiol and ascorbyl tetraisopalmitate synergize to modulate gene expression and restore Hypoxia-Inducible Factor 1 signaling in UV-exposed skin.
Chronic exposure to solar ultraviolet (UV) radiation induces changes to the expression of hundreds of genes in the skin and modulates cellular signaling pathways that alter its structure, function and appearance. To counter these effects, we have developed a 3-in-1 night facial serum (3-in-1 NFS) comprising melatonin, bakuchiol and ascorbyl tetraisopalmitate that is designed to attenuate UV-generated free radicals and support new collagen synthesis. In order to better define its mechanism of action and gain insight into how it might influence the biology of photoaged skin, we performed a transcriptomic analysis of ex vivo skin explants that had been exposed to UV light and treated with 3-in-1 NFS each day for 4 consecutive days. Differentially expressed mRNAs and microRNAs (miRNA) were identified by RNA sequencing and a miRNA interactome was developed. Pathway enrichment analysis was performed to identify pathways likely modulated by 3-in-1 NFS. Our analysis revealed that the combination of active ingredients in 3-in-1 NFS exerted a synergistic effect on skin biology and modulated the expression of genes implicated in the regulation of collagen biosynthesis, angiogenesis, skin barrier function and cellular metabolism. Pathway analysis indicated that these events are driven by Hypoxia-Inducible Factor 1α (HIF-1α) whose expression in UV-exposed skin was partially restored upon 3-in-1 NFS treatment. To our knowledge, 3-in-1 NFS is the first non-drug demonstrated to act upon this pathway in the skin.


How the structure of the large subunit controls function in an oxygen-tolerant [NiFe]-hydrogenase.
Salmonella enterica is an opportunistic pathogen that produces a [NiFe]-hydrogenase under aerobic conditions. In the present study, genetic engineering approaches were used to facilitate isolation of this enzyme, termed Hyd-5. The crystal structure was determined to a resolution of 3.2 Å and the hydro-genase was observed to comprise associated large and small subunits. The structure indicated that His229 from the large subunit was close to the proximal [4Fe-3S] cluster in the small subunit. In addition, His229 was observed to lie close to a buried glutamic acid (Glu73), which is conserved in oxygen-tolerant hydrogenases. His229 and Glu73 of the Hyd-5 large subunit were found to be important in both hydrogen oxidation activity and the oxygen-tolerance mechanism. Substitution of His229 or Glu73 with alanine led to a loss in the ability of Hyd-5 to oxidize hydrogen in air. Furthermore, the H229A variant was found to have lost the overpotential requirement for activity that is always observed with oxygen-tolerant [NiFe]-hydrogenases. It is possible that His229 has a role in stabilizing the super-oxidized form of the proximal cluster in the presence of oxygen, and it is proposed that Glu73could play a supporting role in fine-tuning the chemistry of His229 to enable this function.


Circle quorum system-based non-stop network service model.
Rapid developments in network systems of business service have resulted in more reliance on distributed computing, typified by "subscriber/push" architectures. Unfortunately, frequent and unexpectable network failures were routine, and downtime was not in hours, but in days. High availability has become the most important factor decreasing business risk and improving Quality of Service. Cluster technology has solved the non-stop problem on Local Area Network. However, most technologies including cluster today fail to ensure the non-stop Internet service based on Routers. With good performance on high availability and fault tolerance, quorum systems are very suitable for application to distributed business service networks. In this work, we modeled and developed a non-stop Internet service system based on a new quorum system, circle quorum system, for Boston Mutual Fund Broker, US. With five protocols, it provided highly available data services for clients on Internet.


Prevalence of Dirofilaria immitis infection among shelter cats.
To determine prevalence of Dirofilaria immitis infection among shelter cats. Cross-sectional study. 239 cats euthanatized at an animal shelter in southeastern Michigan. A gross necropsy focusing on the thoracic cavity, heart, lungs, and pulmonary vessels was performed within 5 hours after cats were euthanatized. Blood was collected directly from the heart of cats found to be infected and tested, using a filter test for microfilariae. Serum was tested for D immitis antigens by use of 2 antigen detection kits and for D immitis-specific antibodies by use of 2 antibody detection kits. Cats ranged from approximately 4 months to 15 years old. Adult D immitis were found in 6 (2.5%) cats. Blood could not be recovered from 1 of the cats with heartworm infection. For the 5 other cats, results of the filter test were negative, and results of both antigen and both antibody tests were positive. Results suggest that cats living in an urban area in the northern part of the United States have a low prevalence of adult D immitis infection. However, this is likely to be an underestimate of the true prevalence of infection, because no attempts were made to identify cats infected with larval or juvenile stages of D immitis.


Thermogenesis, low-protein diets, and decreased development of AFB1-induced preneoplastic foci in rat liver.
The development of hepatocellular, putatively preneoplastic, gamma-glutamyl transpeptidase positive (GGT+) foci and tumors induced by aflatoxin B1 (AFB1) has been shown to be reduced in male F344 rats fed a diet containing 6% protein (as casein). This reduction occurs despite increased energy intake, when compared with animals fed a diet containing 22% protein. Among its many effects, low protein intake is known to increase the proportion of energy intake expended in the form of heat (thermogenesis); thus, this investigation examined the association between the development of GGT+ foci and alterations in indices of thermogenesis induced by feeding varying levels of dietary protein. Five days following the completion of AFB1 dosing, animals were assigned to groups fed 4%, 8%, 12%, 16%, or 22% dietary protein for 6 weeks. Foci development (% liver volume occupied) was markedly reduced in animals fed the low-protein diet (4%, 8%), yet calorie consumption per 100 g body wt was greater. A modest negative linear trend was observed in oxygen consumption with increasing levels of dietary protein intake. Urinary norepinephrine levels were elevated in the groups fed 4% and 8% protein; urinary dopamine and norepinephrine turnover rates in brown adipose tissue were highest in animals fed 4% protein. These results suggest that GGT+ foci development occurs when a "critical level" (approx 12%) of dietary protein intake is reached. Inhibition of foci development at lower levels of protein intake is associated with several indicators of increased thermogenesis.


Development of an international comorbidity education framework.
The increasing number of people living with multiple chronic conditions in addition to an index condition has become an international healthcare priority. Health education curricula have been developed alongside single condition frameworks in health service policy and practice and need redesigning to incorporate optimal management of multiple conditions. Our aims were to evaluate current teaching and learning about comorbidity care amongst the global population of healthcare students from different disciplines and to develop an International Comorbidity Education Framework (ICEF) for incorporating comorbidity concepts into health education. We surveyed nursing, medical and pharmacy students from England, India, Italy and Sweden to evaluate their understanding of comorbidity care. A list of core comorbidity content was constructed by an international group of higher education academics and clinicians from the same disciplines, by searching current curricula and analysing clinical frameworks and the student survey data. This list was used to develop the International Comorbidity Education Framework. The survey sample consisted of 917 students from England (42%), India (48%), Italy (8%) and Sweden (2%). The majority of students across all disciplines said that they lacked knowledge, training and confidence in comorbidity care and were unable to identify specific teaching on comorbidities. All student groups wanted further comorbidity training. The health education institution representatives found no specific references to comorbidity in current health education curricula. Current clinical frameworks were used to develop an agreed list of core comorbidity content and hence an International Comorbidity Education Framework. Based on consultation with academics and clinicians and on student feedback we developed an International Comorbidity Education Framework to promote the integration of comorbidity concepts into current healthcare curricula.


The content and clinical utility of psychiatric advance directives.
This paper provides the first systematic examination of the content and clinical utility of psychiatric advance directives, which are documents that specify treatment preferences in advance of periods of compromised decision making. Directives were completed by 106 community mental health center outpatients with at least two psychiatric hospitalizations or emergency department visits within two years. Participants used AD-Maker software in groups of up to six people led by peer trainers. Clinical utility was defined as the degree to which instructions are clinically feasible, useful, and consistent with standards of care. Fifty-five percent of participants were female, and 24 percent were nonwhite. Their mean+/-SD age was 42+/-9.1 years. Primary diagnoses included schizophrenia spectrum disorders (44 percent), bipolar disorders (27 percent), major depression (22 percent), and other disorders (7 percent). Eighty-one percent of participants listed preferred medications, most often antidepressants and second-generation antipsychotics, and 64 percent listed medications they would refuse, most commonly first-generation antipsychotics. Sixty-eight percent preferred hospital alternatives over hospitalization, 89 percent specified methods of de-escalating crises, and 72 percent indicated that they would refuse electroconvulsive therapy (ECT). Forty-six percent appointed a surrogate decision maker. Fifty-seven percent desired a directive that is irrevocable during periods of incapacity. Instructions were rated as feasible, useful, and consistent with practice standards for at least 95 percent of the advance directives, with the exception of instructions about the willingness to use medications not specifically listed in the directive. Results suggested that psychiatric advance directives provide a wealth of treatment preference information that is almost uniformly considered clinically useful. Although the utility of advance directives may vary depending on the circumstances of specific crisis episodes, the information provided can expedite and strengthen clinical care.


MiR-145 negatively regulates Warburg effect by silencing KLF4 and PTBP1 in bladder cancer cells.
The Warburg effect is a well-known feature in cancer-specific metabolism. We previously reported on the role of microRNA (miR)-145 as a tumor-suppressor in human bladder cancer (BC) cells. In this study, we reveal that miR-145 decreases the Warburg effect by silencing KLF4 in BC cells. The expression levels of miR-145 were significantly lower in clinical BC samples and BC cell lines compared to those in normal tissues and HUC cells. Luciferase assay results showed that miR-145 directly bound to 3'UTR of KLF4, which was shown to be overexpressed in the clinical BC samples using Western blot analysis and immunohistochemistry. Remarkable growth inhibition and apoptosis were induced by the ectopic expression of miR-145 or by the gene silencing of KLF4 (siR-KLF4). Also, Warburg effect-related genes such as PTBP1/PKMs were regulated by the transfection of BC cells with miR-145 or siR-KLF4. These results thus indicate that the miR-145/KLF4/PTBP1/PKMs axis is one of the critical pathways that maintain the Warburg effect in BC carcinogenesis. MiR-145 perturbed the Warburg effect by suppressing the KLF4/PTBP1/PKMs pathway in BC cells, resulting in significant cell growth inhibition.


Epigenetics and cell death: DNA hypermethylation in programmed retinal cell death.
Vertebrate genomes undergo epigenetic reprogramming during development and disease. Emerging evidence suggests that DNA methylation plays a key role in cell fate determination in the retina. Despite extensive studies of the programmed cell death that occurs during retinal development and degeneration, little is known about how DNA methylation might regulate neuronal cell death in the retina. The developing chicken retina and the rd1 and rhodopsin-GFP mouse models of retinal degeneration were used to investigate programmed cell death during retinal development and degeneration. Changes in DNA methylation were determined by immunohistochemistry using antibodies against 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC). Punctate patterns of hypermethylation paralleled patterns of caspase3-dependent apoptotic cell death previously reported to occur during development in the chicken retina. Degenerating rd1 mouse retinas, at time points corresponding to the peak of rod cell death, showed elevated signals for 5mC and 5hmC in photoreceptors throughout the retina, with the most intense staining observed in the peripheral retina. Hypermethylation of photoreceptors in rd1 mice was associated with TUNEL and PAR staining and appeared to be cCaspase3-independent. After peak rod degeneration, during the period of cone death, occasional hypermethylation was observed in the outer nuclear layer. The finding that cell-specific increases of 5mC and 5hmC immunostaining are associated with the death of retinal neurons during both development and degeneration suggests that changes in DNA methylation may play a role in modulating gene expression during the process of retinal degeneration. During retinal development, hypermethylation of retinal neurons associates with classical caspase-dependent apoptosis as well as caspase-3 independent cell death, while hypermethylation in the rd1 mouse photoreceptors is primarily associated with caspase-3 independent programmed cell death. These findings suggest a previously unrecognized role for epigenetic mechanisms in the onset and/or progression of programed cell death in the retina.


Sugar as a therapeutic target for the cognitive restoration following traumatic brain injury.
This review aims to discuss examples of changes in glucose (sugar) metabolism after traumatic brain injury (TBI). It will attempt to provide an understanding of what changes in glucose metabolism mean for the injured brain. It will further identify potential therapeutic target(s) emanating from our growing understanding of glucose pathways and their roles in TBI. Although a significant fraction of glucose is utilized for the energy production in the brain, a small fraction is utilized in other, often ignored pathways. Recent studies have unraveled unexpected biological effects of glucose through these pathways, including redox regulation, genetic and epigenetic regulation, glycation of proteins, nucleotide synthesis and amino acid synthesis. A number of regulatory players in minor glucose metabolic pathways, such as folate and chondroitin sulfate proteoglycans, have recently been identified as potential targets to restore cognitive functions. Targeting of these players should be combined with the supplementation of alternative energy substrates to achieve the maximal cognitive restoration after TBI. This multimodal therapeutic strategy deserves testing in various models of TBI. Supplemental digital video content 1: Video that demonstrates an effective therapeutic strategy for the cognitive restoration after TBI. http://links.lww.com/CONR/A46.


Smoking cessation and low-income women: theory, research, and interventions.
There is a growing body of evidence indicating women's experiences with smoking and cessation may be unique from those of men. Advanced practice nurses (APNs) in primary care are ideally suited to assess and intervene with this complex addiction. APNs must be competent in interventions that address cigarette smoking. This article will summarize the literature on smoking cessation interventions with an emphasis on women smokers.


Interactions between host immune response and antigenic variation that control Borrelia burgdorferi population dynamics.
The population dynamics of pathogens within hosts result from interactions between host immune responses and mechanisms of the pathogen to evade or resist immune responses. Vertebrate hosts have evolved adaptive immune responses to eliminate the infection, while many pathogens evade immune clearance through altering surface antigens. Such interactions can result in a characteristic pattern of pathogen population dynamics within hosts consisting of population growth after infection, rapid population decline following specific immune responses, followed by persistence at low densities during a chronic infection stage. Despite the medical importance of chronic infections, little is known about the conditions of the interactions between variable antigens and the adaptive immune system that cause the characteristic pathogen population dynamics. Using the vls antigenic variation system of the Lyme disease pathogen, Borrelia burgdorferi, as a model system, we investigated conditions of the interaction between the antigenic variation system and the adaptive immune response that can explain the within-host population dynamics of B. burgdorferi using mathematical modelling. This characteristic population dynamic pattern can be explained by models that assume a variable immune removal rate of antibody-bound B. burgdorferi. However, models with a constant immune removal rate could reproduce the rapid population decline of B. burgdorferi populations but not their long-term persistence within hosts using parameter values determined by fitting empirical data. The model predictions, along with the assumptions about the interactions between B. burgdorferi and the immune response, can be tested experimentally to estimate the likelihood that each mechanism affects B. burgdorferi population dynamics in real infections.


Association of markers of systemic inflammation, C reactive protein, serum amyloid A, and fibrinogen, with socioeconomic status.
Systemic inflammation may play an important part in the development of cardiovascular disease. It has also been shown that socioeconomic status predicts cardiovascular events independently of established risk factors. The aim of this study was to analyse the association of three sensitive markers of systemic inflammation: C reactive protein (CRP), serum amyloid A protein (SAA), and fibrinogen, with socioeconomic status. Cross sectional study. Eastern and southern Finland. 1503 men aged 45 to 74 years who participated in a cardiovascular risk factor survey in 1997. Based on the levels of education and family income, the men were classified to three socioeconomic groups. Mean concentrations of CRP (p for the trend <0.001), SAA (p for the trend 0.018), and fibrinogen (p for the trend <0.001) decreased substantially with increasing socioeconomic status. The trends in CRP and fibrinogen remained statistically significant after adjustment for smoking, waist to hip ratio, and prevalent longstanding diseases, and a non-significant trend was found for SAA (p for the trend 0.118). The inverse association between inflammation markers and socioeconomic status was particularly strong among the men below 60 years of age. Systemic inflammation is a potential mediator, especially among young and middle aged men, for the association between socioeconomic status and cardiovascular disease.


BPI-ANCA and long-term prognosis among 46 adult CF patients: a prospective 10-year follow-up study.
Anti-neutrophil cytoplasmic antibodies specific for bactericidal/permeability-increasing protein (BPI-ANCA) are frequent in CF patients and mainly develop in response to infection with Pseudomonas aeruginosa. It is not known to what extent BPI-ANCA correlates to prognosis. To evaluate the prognostic value of IgA-BPI-ANCA, measured at the beginning of the study, for transplantation-free survival. A cohort of 46 adult, nontransplanted CF patients was generated, 1995-1998, and characterized using Leeds criteria, lung function, and IgA-BPI-ANCA levels measured by ELISA. The cohort was followed until December 2009, using the combined endpoint of death or lung transplantation. Lung function and IgA-BPI-ANCA, but not Leeds criteria, were significantly associated with adverse outcome. No patient with normal lung function at baseline reached endpoint. Within 10 years 8/11 with high BPI-ANCA reached an endpoint compared to 3/17 ANCA-negative patients. A similar result was seen within the Leeds I group where 7 out of 9 BPI-ANCA-positive patients reached endpoint, compared to none of the 5 patients without BPI-ANCA. IgA-BPI-ANCA is associated with adverse outcome among Pseudomonas aeruginosa infected CF patients, suggesting that BPI-ANCA is a biomarker of an unfavourable host-pathogen interaction.


Enhancing the chelation capacity of rice to maximise iron and zinc concentrations under elevated atmospheric carbon dioxide.
Roughly half of the Earth's seven billion people rely on rice as their primary source of food. The milled grain of rice, often referred to as polished or white rice, serves as a rich source of energy but is low in protein and several essential micronutrients such as iron and zinc. As a result, billions of people in rice-based countries suffer the debilitating effects of protein-energy and micronutrient malnutrition with symptoms including iron-deficiency anaemia, growth retardation and blindness. By 2050, the Earth's atmospheric carbon dioxide concentration ([CO2]) is expected to reach 550μmolmol-1, representing a 70% increase from today's concentration of 392μmolmol-1. The impacts of elevated [CO2] on plant growth will likely include agronomically useful traits such as increased biomass, yield and water-use efficiency. However, increased plant productivity is likely to be accompanied by decreased protein and micronutrient mineral concentrations of cereal grain. This review focuses on the effects of carbon dioxide-enrichment on rice physiology and nutritional composition and proposes increased activity of the Strategy II iron uptake pathway as a promising method to maintain or increase iron and zinc concentrations in rice grain, and perhaps cereal grain in general, under elevated [CO2].


Safety and pharmacokinetics of ciprofloxacin dry powder for inhalation in cystic fibrosis: a phase I, randomized, single-dose, dose-escalation study.
Reliable, reproducible deposition to the lung is a major prerequisite for the clinical use of inhaled drugs. Ciprofloxacin dry powder for inhalation (ciprofloxacin DPI; Bayer HealthCare AG, Leverkusen, Germany) is an antibacterial therapy in development using Novartis' PulmoSphere™ technology (Novartis Pharma AG, Basel, Switzerland) for the targeted delivery of ciprofloxacin to the lung via a T-326 inhaler. This randomized, single-blind, placebo-controlled, dose-escalation study investigated the safety, tolerability, and pharmacokinetics of single-dose ciprofloxacin DPI (32.5 mg [n=6] or 65 mg [n=6]) and matching placebo (n=4) in adult patients with cystic fibrosis and stable pulmonary status (forced expiratory volume in 1 sec ≥30%) who were colonized with Pseudomonas aeruginosa. Peak sputum concentrations of 34.9 mg/L (range 2.03-229) and 376 mg/L (8.95-1283) for ciprofloxacin 32.5 mg and 65 mg, respectively, indicated targeting of ciprofloxacin DPI to the lung. This contrasted with low systemic exposure (peak plasma concentrations: 0.0790 mg/L [32.5 mg] and 0.182 mg/L [65 mg]). Single-dose ciprofloxacin DPI 32.5 mg or 65 mg was well tolerated with similar incidences of adverse events across all groups. No deaths, discontinuations, treatment-related serious adverse events, or clinically relevant changes in laboratory parameters, vital signs, or lung function tests were reported. Lung targeting with high pulmonary concentrations of ciprofloxacin combined with low systemic exposure was confirmed. These results support further study of ciprofloxacin DPI as a potentially more convenient alternative to nebulized antibiotic solutions for managing chronic lung infections.


[Changes in the hormonal function of the hypophysis-adrenal system by administration of cortisol in 'fixed' phases of the postnatal ontogenesis in rats].
Administration of cortisol on the 13th, 15th, and 17th days of postnatal ontogenesis was shown to exert a long-lasting modifying effect upon the hypophysis-adrenal system's (HAS) stress reactivity in the rat litter. The effect involved a longer duration of hormonal response to immobilisation stress: a longer-lasting elevation of the corticosterone level in the blood plasma suggesting a diminished sensitivity of the HAS to feed-back signal. In hippocampus, frontal cortex and hypothalamus of the experimental rats, a decrease in receptor binding of labeled corticosterone was revealed.


Transient receptor potential melastatin 2 protects mice against polymicrobial sepsis by enhancing bacterial clearance.
Recent studies suggest that the transient receptor potential melastatin 2 (TRPM2) channel plays an important role in inflammation and immune response. However, the role and mechanism of TRPM2 in polymicrobial sepsis remain unclear. The authors explored the effects of genetic disruption of TRPM2 on mortality (n = 15), bacterial clearance (n = 6), organ injury, and systemic inflammation during cecal ligation and puncture-induced sepsis. Electrophysiology, immunoblot, bacterial clearance experiment, and quantitative real-time polymerase chain reaction were used to explore the role and mechanism of TRPM2 in sepsis. After cecal ligation and puncture, Trpm2-knockout mice had increased mortality compared with wild-type mice (73.3 vs. 40%, P = 0.0289). The increased mortality was associated with increased bacterial burden, organ injury, and systemic inflammation. TRPM2-mediated Ca influx plays an important role in lipopolysaccharide or cecal ligation and puncture-induced heme oxygenase-1 (HO-1) expression in macrophage. HO-1 up-regulation decreased bacterial burden both in wild-type bone marrow-derived macrophages and in cecal ligation and puncture-induced septic wild-type mice. Disruption of TRPM2 decreased HO-1 expression and increased bacterial burden in bone marrow-derived macrophages. Pretreatment of Trpm2-knockout bone marrow-derived macrophages with HO-1 inducer markedly increased HO-1 expression and decreased bacterial burden. Pretreatment of Trpm2-knockout mice with HO-1 inducer reversed the susceptibility of Trpm2-knockout mice to sepsis by enhancing the bacterial clearance. In addition, septic patients with lower monocytic TRPM2 and HO-1 messenger RNA levels had a worse outcome compared with septic patients with normal monocytic TRPM2 and HO-1 messenger RNA levels. TRPM2 levels correlated with HO-1 levels in septic patients (r = 0.675, P = 0.001). The study data demonstrate a protective role of TRPM2 in controlling bacterial clearance during polymicrobial sepsis possibly by regulating HO-1 expression.


IFITMs from Mycobacteria Confer Resistance to Influenza Virus When Expressed in Human Cells.
Interferon induced transmembrane proteins (IFITMs) found in vertebrates restrict infections by specific viruses. IFITM3 is known to be essential for restriction of influenza virus infections in both mice and humans. Vertebrate IFITMs are hypothesized to have derived from a horizontal gene transfer from bacteria to a primitive unicellular eukaryote. Since bacterial IFITMs share minimal amino acid identity with human IFITM3, we hypothesized that examination of bacterial IFITMs in human cells would provide insight into the essential characteristics necessary for antiviral activity of IFITMs. We examined IFITMs from Mycobacterium avium and Mycobacterium abscessus for potential antiviral activity. Both of these IFITMs conferred a moderate level of resistance to influenza virus in human cells, identifying them as functional homologues of IFITM3. Analysis of sequence elements shared by bacterial IFITMs and IFITM3 identified two hydrophobic domains, putative S-palmitoylation sites, and conserved phenylalanine residues associated with IFITM3 interactions, which are all necessary for IFITM3 antiviral activity. We observed that, like IFITM3, bacterial IFITMs were S-palmitoylated, albeit to a lesser degree. We also demonstrated the ability of a bacterial IFITM to co-immunoprecipitate with IFITM3 suggesting formation of a complex, and also visualized strong co-localization of bacterial IFITMs with IFITM3. However, the mycobacterial IFITMs lack the endocytic-targeting motif conserved in vertebrate IFITM3. As such, these bacterial proteins, when expressed alone, had diminished colocalization with cathepsin B-positive endolysosomal compartments that are the primary site of IFITM3-dependent influenza virus restriction. Though the precise evolutionary origin of vertebrate IFITMs is not known, our results support a model whereby transfer of a bacterial IFITM gene to eukaryotic cells may have provided a selective advantage against viral infection that was refined through the course of vertebrate evolution to include more robust signals for S-palmitoylation and localization to sites of endocytic virus trafficking.


Spatially resolved confocal resonant Raman microscopic analysis of anode-grown Geobacter sulfurreducens biofilms.
When grown on the surface of an anode electrode, Geobacter sulfurreducens forms a multi-cell thick biofilm in which all cells appear to couple the oxidation of acetate with electron transport to the anode, which serves as the terminal metabolic electron acceptor. Just how electrons are transported through such a biofilm from cells to the underlying anode surface over distances that can exceed 20 microns remains unresolved. Current evidence suggests it may occur by electron hopping through a proposed network of redox cofactors composed of immobile outer membrane and/or extracellular multi-heme c-type cytochromes. In the present work, we perform a spatially resolved confocal resonant Raman (CRR) microscopic analysis to investigate anode-grown Geobacter biofilms. The results confirm the presence of an intra-biofilm redox gradient whereby the probability that a heme is in the reduced state increases with increasing distance from the anode surface. Such a gradient is required to drive electron transport toward the anode surface by electron hopping via cytochromes. The results also indicate that at open circuit, when electrons are expected to accumulate in redox cofactors involved in electron transport due to the inability of the anode to accept electrons, nearly all c-type cytochrome hemes detected in the biofilm are oxidized. The same outcome occurs when a comparable potential to that measured at open circuit (-0.30 V vs. SHE) is applied to the anode, whereas nearly all hemes are reduced when an exceedingly negative potential (-0.50 V vs. SHE) is applied to the anode. These results suggest that nearly all c-type cytochrome hemes detected in the biofilm can be electrochemically accessed by the electrode, but most have oxidation potentials too negative to transport electrons originating from acetate metabolism. The results also reveal a lateral heterogeneity (x-y dimensions) in the type of c-type cytochromes within the biofilm that may affect electron transport to the electrode.


Artificial neural networks as supervised techniques for FT-IR microspectroscopic imaging.
In this report the applicability of an improved method of image segmentation of infrared microspectroscopic data from histological specimens is demonstrated. Fourier transform infrared (FT-IR) microspectroscopy was used to record hyperspectral data sets from human colorectal adenocarcinomas and to build up a database of spatially resolved tissue spectra. This database of colon microspectra comprised 4120 high-quality FT-IR point spectra from 28 patient samples and 12 different histological structures. The spectral information contained in the database was employed to teach and validate multilayer perceptron artificial neural network (MLP-ANN) models. These classification models were then employed for database analysis and utilised to produce false colour images from complete tissue maps of FT-IR microspectra. An important aspect of this study was also to demonstrate how the diagnostic sensitivity and specificity can be specifically optimised. An example is given which shows that changes of the number of teaching patterns per class can be used to modify these two interrelated test parameters. The definition of ANN topology turned out to be crucial to achieve a high degree of correspondence between the gold standard of histopathology and IR spectroscopy. Particularly, a hierarchical scheme of ANN classification proved to be superior for the reliable classification of tissue spectra. It was found that unsupervised methods of clustering, specifically agglomerative hierarchical clustering (AHC), were helpful in the initial phases of model generation. Optimal classification results could be achieved if the class definitions for the ANNs were carried out by considering the classification information provided by cluster analysis.


[Effect of sympathectomy on the heart pump function in rats during postnatal ontogenesis].
The influence of guanetidine sympathectomy (30 mg/kg) on the heart pump function in rats during 3 weeks in postnatal ontogenesis has been investigated. Sympathectomy restrains age-dependent establishment of stroke volume, cardiac output and heart rate. The adaptation effects of regular physical training do not develop in the animals with sympathectomy, i.e. heart rate does not decrease and stroke volume does not increase. The initial stage of adaptation of the sympathectomized animals to physical training is accompanied by decrease in stroke volume and remarkable increase in heart rate which indicates the reduction of contractile activity in the myocardium.


Long-term survival analysis of the Canadian Lung Volume Reduction Surgery trial.
The Canadian Lung Volume Reduction Surgery (CLVRS) trial was a multicentered randomized controlled trial that concluded that lung volume reduction surgery improves functional status and health-related quality of life (for at least 2 years) in selected patients with advanced emphysema. This retrospective observational study assessed the long-term survival of patients enrolled in the CLVRS at 8 to 10 years after randomization. Vital statistics were gathered through telephone contact, physician records, and municipal obituaries. Survival analysis was undertaken using Kaplan-Meier and the Cox proportional hazards models. Fifty-two patients (84% of the initial 62 patients randomized) had a median survival time of 4.11 years. A 16-month survival advantage and a 20% reduction in mortality was observed in the LVRS group as compared with the best medical care group. Although clinically meaningful, these differences were not statistically significant. These findings echo those of other published reports and demonstrate the potential long-term benefit of LVRS in the treatment of end-stage emphysema.


Amino acid derangements in adults with severe falciparum malaria.
Amino acid derangements are common in severe falciparum malaria and have been associated with endothelial dysfunction (L-arginine), metabolic acidosis (alanine and lactate), and disease severity (phenylalanine and tryptophan metabolites). Whether these amino acid perturbations reflect isolated pathogenic mechanisms or if they are part of overall changes in amino acid metabolism is unclear. To investigate this, we prospectively simultaneously quantified a broad range of plasma free amino acids (PFAA) using HPLC-MRM-Mass spectrometry in relation to presenting symptoms in adults with severe malaria (n = 88), septicaemia (n = 88), uncomplicated malaria (n = 71), and healthy controls (n = 48) from Bangladesh. The total plasma concentration of measured amino acids was significantly reduced in each of the patient groups when compared to normal levels observed in healthy local controls: uncomplicated malaria -54%, severe malaria -23%, and sepsis -32%, (p = <0.001). Inspection of amino acid profiles revealed that in each group the majority of amino acids were below normal levels, except for phenylalanine. Among patients with severe malaria, L-lactate was strongly associated with an increase of the total amino acid concentration, likely because this reflects tissue hypoxia. Our data confirm previously described amino acid abnormalities, likely resulting from overall changes in the concentration of PFAA.


Applications of the experience sampling method (ESM) in paediatric healthcare: a systematic review.
With the Experience Sampling Method (ESM) participants are asked to provide self-reports of their symptoms, feelings, thoughts and behaviours in daily life. This preregistered systematic review assessed how ESM is being used to monitor emotional well-being, somatic health, fatigue and pain in children and adolescents with a chronic somatic illness. Databases were searched from inception. Studies were selected if they included children or adolescents aged 0-25 years with a chronic somatic illness and used ESM focussing on mental health or psychosocial wellbeing, biopsychosocial factors and/or somatic health. Two reviewers extracted data of the final 47 papers, describing 48 studies. Most studies evaluated what factors influence medical or psychological symptoms and how symptoms influence each other. Another common purpose was to study the feasibility of ESM or ESM as part of an app or intervention. Study methods were heterogeneous and most studies lack adequate reporting of ESM applications and results. While ESM holds great potential for providing results and feedback to patients and caregivers, little use is being made of this option. Future studies should consider what they report in their studies, conduct a priori power analyses and how ESM can be embedded in clinical practice. While ESM has many clinical applications, it is currently mostly used for research purposes. Current studies using ESM are heterogeneous and lack consistent, high-quality reporting. There is great potential in ESM for providing patients and parents with personalised feedback.


Local mitochondrial-endolysosomal microfusion cleaves voltage-dependent anion channel 1 to promote survival in hypoxia.
The oxygen-limiting (hypoxic) microenvironment of tumors induces metabolic reprogramming and cell survival, but the underlying mechanisms involving mitochondria remain poorly understood. We previously demonstrated that hypoxia-inducible factor 1 mediates the hyperfusion of mitochondria by inducing Bcl-2/adenovirus E1B 19-kDa interacting protein 3 and posttranslational truncation of the mitochondrial ATP transporter outer membrane voltage-dependent anion channel 1 in hypoxic cells. In addition, we showed that truncation is associated with increased resistance to drug-induced apoptosis and is indicative of increased patient chemoresistance. We now show that silencing of the tumor suppressor TP53 decreases truncation and increases drug-induced apoptosis. We also show that TP53 regulates truncation through induction of the mitochondrial protein Mieap. While we found that truncation was independent of mitophagy, we observed local microfusion between mitochondria and endolysosomes in hypoxic cells in culture and in patients' tumor tissues. Since we found that the endolysosomal asparagine endopeptidase was responsible for truncation, we propose that it is a readout of mitochondrial-endolysosomal microfusion in hypoxia. These novel findings provide the framework for a better understanding of hypoxic cell metabolism and cell survival through mitochondrial-endolysosomal microfusion regulated by hypoxia-inducible factor 1 and TP53.


Malignant Pleural Effusions-A Window Into Local Anti-Tumor T Cell Immunity?
The success of immunotherapy that targets inhibitory T cell receptors for the treatment of multiple cancers has seen the anti-tumor immune response re-emerge as a promising biomarker of response to therapy. Longitudinal characterization of T cells in the tumor microenvironment (TME) helps us understand how to promote effective anti-tumor immunity. However, serial analyses at the tumor site are rarely feasible in clinical practice. Malignant pleural effusions (MPE) associated with thoracic cancers are an abnormal accumulation of fluid in the pleural space that is routinely drained for patient symptom control. This fluid contains tumor cells and immune cells, including lymphocytes, macrophages and dendritic cells, providing a window into the local tumor microenvironment. Recurrent MPE is common, and provides an opportunity for longitudinal analysis of the tumor site in a clinical setting. Here, we review the phenotype of MPE-derived T cells, comparing them to tumor and blood T cells. We discuss the benefits and limitations of their use as potential dynamic biomarkers of response to therapy.


Association between sociodemographic, behavioral, and medical conditions and signs of temporomandibular disorders across gender: results of the study of health in Pomerania (SHIP-0).
To examine whether there is a gender-dependent risk profile for signs of temporomandibular disorders (TMD) in a population-based sample. Sociodemographic, behavioral, and medical factors were checked for associations with TMD in a cross-sectional study of 3,567 subjects aged 25 to 74 years in Germany. Data were collected from clinical examinations, interviews, and questionnaires. Logistic regression analyses were used to estimate factors associated with signs of TMD across gender. TMD signs included tenderness or pain on palpation of 3 or more masticatory muscles and tenderness or pain on palpation in 1 or both temporomandibular joints (TMJs). In women, muscle tenderness or pain was found to be significantly associated with general arthrosis/arthritis and lower back pain. In men, muscle tenderness or pain was significantly associated with school education > 11 years, various categories of loss of occlusal support, lip/tongue/cheek biting, and general arthrosis/arthritis. In women, TMJ tenderness or pain was associated with widowed status, bruxism, general arthrosis/arthritis, lower back pain, and sex-hormone replacement. In men, TMJ tenderness or pain was associated with multiple losses of posterior supporting zones, gout, and lower back pain. In women, there were inverse associations between loss of occlusal support in 3 posterior zones and muscle and TMJ tendernes. Except for some general health conditions and bruxism, the hypothesis of a gender-dependent risk profile for signs of TMD is partly supported. The results of this study indicate that TMD is a complex disorder associated with mixed etiologic factors between genders.


Reinforcement Learning in Spiking Neural Networks with Stochastic and Deterministic Synapses.
Though succeeding in solving various learning tasks, most existing reinforcement learning (RL) models have failed to take into account the complexity of synaptic plasticity in the neural system. Models implementing reinforcement learning with spiking neurons involve only a single plasticity mechanism. Here, we propose a neural realistic reinforcement learning model that coordinates the plasticities of two types of synapses: stochastic and deterministic. The plasticity of the stochastic synapse is achieved by the hedonistic rule through modulating the release probability of synaptic neurotransmitter, while the plasticity of the deterministic synapse is achieved by a variant of a reward-modulated spike-timing-dependent plasticity rule through modulating the synaptic strengths. We evaluate the proposed learning model on two benchmark tasks: learning a logic gate function and the 19-state random walk problem. Experimental results show that the coordination of diverse synaptic plasticities can make the RL model learn in a rapid and stable form.


Conserved polar residues in the transmembrane domain of the human tachykinin NK2 receptor: functional roles and structural implications.
We have studied the effects of agonist and antagonist binding, agonist-induced activation and agonist-induced desensitization of the human tachykinin NK2 receptor mutated at polar residues Asn-51 [in transmembrane helix 1 (TM1)], Asp-79 (TM2) and Asn-303 (TM7), which are highly conserved in the transmembrane domain in the rhodopsin family of G-protein-coupled receptors. Wild-type and mutant receptors were expressed in both COS-1 cells and Xenopus oocytes. The results show that the N51D mutation results in a receptor which, in contrast with the wild-type receptor, is desensitized by the application of a concentration of 1 microM of the partial agonist GR64349, indicating that the mutant is more sensitive to agonist activation than is the wild-type receptor. In addition, we show that, whereas the D79E mutant displayed activation properties similar to those of the wild-type receptor, the D79N and D79A mutants displayed a severely impaired ability to activate the calcium-dependent chloride current. This suggests that it is the negative charge at Asn-79, rather than the ability of this residue to hydrogen-bond, that is critical for the activity of the receptor. Interestingly, the placement of a negative charge at position 303 could compensate for the removal of the negative charge at position 79, since the double mutant D79N/N303D displayed activation properties similar to those of the wild-type receptor. This suggests that these two residues are functionally coupled, and may even be in close proximity in the three-dimensional structure of the human tachykinin NK2 receptor. A three-dimensional model of the receptor displaying this putative interaction is presented.


Secretory proteins induced in human fibroblasts under conditions used for the production of interferon beta.
Human fibroblast cells treated with a combination of inhibitors of protein and RNA synthesis [cycloheximide and actinomycin D as used to superinduce interferon beta (IFN-beta)] secrete two proteins with molecular masses of 22000 and 27000 kilodaltons (called 22-kDal and 27-kDal) that are precipitable with an antiserum raised against impure IFN-beta but are antigenically distinct from IFN-beta 1. Translation in vitro of mRNA extracted from human fibroblast cells induced for the production of IFN-beta leads to the synthesis of a 26-kDal protein that is structurally closely related to the 22- and 27-kDal proteins. This 26-kDal protein mRNA is relatively abundant and also appears in human fibroblasts induced only with cycloheximide. It has been partially purified by sucrose gradient centrifugation and more extensively by diazobenzyloxymethyl-cellulose hybridization to plasmid DNA from a bacterial cDNA clone. When translated in an in vitro reticulocyte system supplemented with dog pancreas microsomes, the 26-kDal protein and two other intermediates corresponding presumably to its signal-cleaved (19-kDal) and partially glycosylated (24-kDal) forms were observed. Crude, partially purified, and highly purified 26-kDal mRNA failed to program the synthesis of antiviral or ppp(A2'p5')nA synthetase-inducing activity when translated in Xenopus laevis oocytes. Moreover, partially purified 22-kDal and 27-kDal (i.e., the in vivo equivalents of the 26-kDal protein) are also devoid of antiviral or ppp(A2'p5')nA synthetase-inducing activity. Hence, this 26-kDal mRNA, although presumably identical to the human IFN-beta 2 mRNA described by Weissenbach et al. [Weissenbach, J., Chernajovsky, Y., Zeevi, M., Shulman, K., Soreq, H., Nir. U., Wallach, D., Perricaudet, M., Tiollais, P. & Revel, M. (1980) Proc. Natl. Acad. Sci. USA 77, 7152-7156], cannot be considered to be a fibroblast interferon mRNA.


LG839: anti-obesity effects and polymorphic gene correlates of reward deficiency syndrome.
This study systematically assessed the weight management effects of a novel experimental DNA-customized nutraceutical, LG839 (LifeGen, Inc., La Jolla, CA, USA). A total of 1058 subjects who participated in the overall D.I.E.T. study were genotyped and administered an LG839 variant based on polymorphic outcomes. A subset of 27 self-identified obese subjects of Dutch descent, having the same DNA pattern of four out of the five candidate genes tested (chi-square analysis) as the entire data set, was subsequently evaluated. Simple t tests comparing a number of weight management parameters before and after 80 days of treatment with LG839 were performed. Significant results were observed for weight loss, sugar craving reduction, appetite suppression, snack reduction, reduction of late night eating (all P<0.01), increased perception of overeating, enhanced quality of sleep, increased happiness (all P<0.05), and increased energy (P<0.001). Polymorphic correlates were obtained for a number of genes (LEP, PPAR-gamma2, MTHFR, 5-HT2A, and DRD2 genes) with positive clinical parameters tested in this study. Of all the outcomes and gene polymorphisms, only the DRD2 gene polymorphism (A1 allele) had a significant Pearson correlation with days on treatment (r=0.42, P=0.045). If these results are confirmed in additional rigorous, controlled studies, we carefully suggest that DNA-directed targeting of certain regulator genes, along with customized nutraceutical intervention, provides a unique framework and strategic modality to combat obesity.


Up- and down-regulation of granulocyte/macrophage-colony stimulating factor activity in murine skin increase susceptibility to skin carcinogenesis by independent mechanisms.
The role of granulocyte-macrophage colony-stimulating factor (GM-CSF) in tumorigenesis is complex. On the one hand, GM-CSF can promote tumor cell growth, survival, and even metastasis. On the other hand, it can stimulate tumor cell rejection. In skin, it is early expressed after topic application of tumor-promoting agents and therefore may be responsible for changes that correlate with skin tumor promotion (e.g., epidermal hyperproliferation and inflammation). To analyze GM-CSF function in skin tumorigenesis, we generated transgenic mice epidermally overexpressing either GM-CSF or a GM-CSF antagonist. Both types of transgenic mice exhibited significantly increased numbers of benign tumors in a two-step skin carcinogenesis experiment using 7',12'-dimethylbenz[a]anthracene (DMBA) as initiator and 12-O-tetradecanoylphorbol-CSF displayed a significantly elevated carcinoma burden following a single-step carcinogenesis protocol consisting of tumor initiation only. Therefore, endogenous promotion is responsible for elevated tumor development in GM-CSF-overexpressing mice. In antagonist transgenic animals, an increased tumorigenicity of modified B16 tumor cells after cutaneous transplantation as compared with nontransgenic or GM-CSF transgenic mice was observed. Thus, the antitumor activity leading to the repression of tumor cell growth in control mice is GM-CSF dependent and is compromised in mice expressing the antagonist. We suggest that both, up-regulation and down-regulation of GM-CSF activity in skin, increase the incidence and growth of tumors via two independent mechanisms: endogenous tumor promotion in the case of increased GM-CSF activity and compromised tumor cell rejection in the case of decreased GM-CSF activity.


Everyday and prospective memory deficits in ecstasy/polydrug users.
The impact of ecstasy/polydrug use on real-world memory (i.e. everyday memory, cognitive failures and prospective memory [PM]) was investigated in a sample of 42 ecstasy/polydrug users and 31 non-ecstasy users. Laboratory-based PM tasks were administered along with self-reported measures of PM to test whether any ecstasy/polydrug-related impairment on the different aspects of PM was present. Self-reported measures of everyday memory and cognitive failures were also administered. Ecstasy/polydrug associated deficits were observed on both laboratory and self-reported measures of PM and everyday memory. The present study extends previous research by demonstrating that deficits in PM are real and cannot be simply attributed to self-misperceptions. The deficits observed reflect some general capacity underpinning both time- and event-based PM contexts and are not task specific. Among this group of ecstasy/polydrug users recreational use of cocaine was also prominently associated with PM deficits. Further research might explore the differential effects of individual illicit drugs on real-world memory.


Programmable mechanically assisted geometric deformations of glassy two-stage reactive polymeric materials.
Thiol-isocyanate-methacrylate two-stage reactive network polymers were developed and used for fabrication of well-defined surface patterns as well as functional geometric shapes to demonstrate a new methodology for processing of "smart materials". The dynamic stage I networks were synthesized in base-catalyzed thiol-isocyanate cross-linking reactions to yield tough, glassy materials at ambient conditions. Methacrylate-rich stage I networks, incorporating photoinitiator and photoabsorber, were irradiated with UV light to generate stage II networks with intricate property gradients. Upon directional straining and subsequent temperature-dependent stress relief of the predefined gradient regions, the desired surface or bulk geometric transformations were achieved. Depending on the gradient extent in conjunction with photoorthogonal initiators, the introduced deformations were shown to be easily erasable by heat or permanently fixable by bulk polymerization.


Chemical ordering in Al(72)Ni(20)Co(8) decagonal quasicrystals.
First-principles total energy calculations of the 2-nm clusters seen in high-perfection Al (72)Ni(20)Co(8) decagonal quasicrystals demonstrate that chemical ordering between Al and transition metals in the central ring is energetically highly favorable. The chemical ordering introduces extensive structure relaxation and results in broken decagonal symmetry. Such broken symmetry is sufficient to enforce the perfect quasiperiodic tiling.


Sprouting of noradrenergic fibers in hippocampus after medial septal lesions: contributions of the central and peripheral nervous systems.
The neuronal sprouting of noradrenergic fibers was studied in the hippocampal formation. The extent and time course of lesion-induced plasticity of both central and peripheral noradrenergic neurons was determined by assaying norepinephrine (NE) concentrations and high-affinity [3H]NE uptake in the dentate gyrus at 2 to 16 weeks after medial septal lesions. Two weeks after a medial septal lesion there was a dramatic decrease in dentate NE. During the subsequent weeks normal concentrations of dentate NE were reestablished. The recovery or increase of NE with time reflected a contribution from both central and peripheral noradrenergic systems. Although both central and peripheral noradrenergic systems contributed to this recovery, they did so in very different ways. The time course of the central noradrenergic response was slower than that of the peripheral system and the final NE concentrations were quite different for the two systems. The central adrenergic system's sprouting response apparently stabilized when normal NE concentrations were attained, whereas the ingrowth of peripheral sympathetic fibers continued to concentrations that were well above normal unoperated levels. The findings have implications in relationship to the different possible controlling mechanisms governing neuronal plasticity of the central and peripheral noradrenergic systems.


Genetic diversity of Pneumocystis jirovecii isolates among Turkish population based on mitochondrial large subunit ribosomal RNA and dihydropteroate synthase gene typing.
Pneumocystis jirovecii (P. jirovecii) is an atypical fungus that can cause severe interstitial pneumonia in immunocompromised patients. In this study, mitochondrial large subunit ribosomal RNA (mtLSU-rRNA) and dihydropteroate synthase (DHPS) gene polymorphism in P. jirovecii isolates were investigated in Western Turkey's Izmir province and its surroundings. For this purpose, a total of 157 P. jirovecii isolates obtained from bronchoalveolar lavage samples of hospitalized cases and lung tissue samples of autopsy cases who died outside hospital were examined. Genotypes were identified by direct sequencing of mtLSU-rRNA restriction fragment length polymorphism analysis of the DHPS gene amplicons. The mtLSU-rRNA analysis revealed that genotype 2 was the most common genotype with 58%. The following genotypes were genotype 3 (13%), genotype 1 (11.6%) and genotype 4 (5.1%), while genotype 5 (0.7%) was detected in only one autopsy case. In addition, 16 (11.6%) cases had dual or triple different genotypes (mixed infection). It was observed that the genotype distribution was not affected by characteristics such as age, gender and immune status. However, the predominance of genotype 2 in solid organ tumors and the predominance of mixed infection in patients with chronic pulmonary disease were statistically significant. On the other hand, DHPS gene amplification was positive in 137 (87.3%) of 157 samples. While no mutation was observed in 135 samples, the association of wild-type and 57th codon mutation was detected in two hospitalized cases (1.5%). In this study, important epidemiological data on the distribution of mtLSU-rRNA genotypes were obtained. Also the existence of DHPS gene mutations associated with potential drug resistance in our community was shown for the first time. Further studies are needed to evaluate the possible effects of genotypes on the prognosis of the disease to help with the clinician's treatment decisions. Pneumocystis jirovecii (P. jirovecii) is an atypical fungus that can cause life-threatening pneumonia in immunocompromised patients. In this study, we investigated the mtLSU-rRNA and DHPS gene polymorphisms in P. jirovecii isolates from both hospital and autopsy cases.


Mechanical behavior of articular cartilage quantitative changes with enzymatic alteration of the proteoglycan fraction.
The in-vitro viscoelastic mechanical response of normal rabbit articular cartilage is strongly dependent on the quantity and integrity of the proteoglycan fraction of the tissue matrix. Experimental results demonstrate that specific functional relationships exist between shear moduli, retardation time spectra, and proteoglycan content. Quantitative enzymolysis of the proteoglycan fraction of the tissue alters the form of these relationships in a fashion consistent with the altered physiochemical make-up of the tissue. The observed changes in mechanical behavior with controlled enzymolysis are similar to those associated with the early stages of osteoarthritis, rheumatoid arthritis, joint sepsis, and synovitis in animal models.


Vitamin A and Retinoids as Mitochondrial Toxicants.
Vitamin A and its derivatives, the retinoids, are micronutrient necessary for the human diet in order to maintain several cellular functions from human development to adulthood and also through aging. Furthermore, vitamin A and retinoids are utilized pharmacologically in the treatment of some diseases, as, for instance, dermatological disturbances and some types of cancer. In spite of being an essential micronutrient with clinical application, vitamin A exerts several toxic effects regarding redox environment and mitochondrial function. Moreover, decreased life quality and increased mortality rates among vitamin A supplements users have been reported. However, the exact mechanism by which vitamin A elicits its deleterious effects is not clear yet. In this review, the role of mitochondrial dysfunction in the mechanism of vitamin A-induced toxicity is discussed.


The case for an ancestral genetic system involving simple analogues of the nucleotides.
The idea that the first living systems on earth were based on self-replicating RNA molecules has recently become popular as a result of the discovery of ribozymes. However, there are several major problems associated with the prebiotic synthesis of ribonucleotides. In addition, there is the newly recognized problem of enantiomeric cross-inhibition, whereby template-directed polymerization involving one enantiomer of RNA is inhibited strongly by the presence of the other enantiomer. Here we propose that RNA was preceded in the evolution of life by a polymer constructed from flexible, acyclic, probably prochiral nucleotide analogues that were synthesized readily on the primitive earth. Several potentially prebiotic nucleotide analogues are considered in this context, and some of the consequences of this proposal are discussed.


Factors that prompted families to file medical malpractice claims following perinatal injuries.
To identify self-reported reasons that prompt families to file malpractice claims following perinatal injuries. Families were interviewed by telephone using a questionnaire that contained structured and open-ended questions. Mothers of infants who had experienced permanent injuries or deaths and had closed malpractice claims in Florida between 1986 and August 1989 were interviewed. Questionnaires were completed by 127 (35%) of a total of 368 such families. Reasons prompting families to file and families' descriptions of medical events, advice from acquaintances, and the quality of physician-family communication. Families volunteered numerous reasons for filing: advised by knowledgeable acquaintances (33% of respondents), recognized cover-up (24%), needed money (24%), recognized that their child would have no future (23%), needed information (20%), and decided to seek revenge or protect others from harm (19%). Over one third of all families indicated that they were told by medical personnel prior to filing that the care provided had caused their children's injuries. Families expressed dissatisfaction with physician-patient communication. Families believed that physicians would not listen (13% of sample), would not talk openly (32%), attempted to mislead them (48%), or did not warn about long-term neurodevelopmental problems (70%). Families give many reasons for filing a claim. Obtaining money may not be the only goal for some families who file suit.


Harsh Parenting As a Potential Mediator of the Association Between Intimate Partner Violence and Child Disruptive Behavior in Families With Young Children.
Young children living with intimate partner violence (IPV) are often also exposed to harsh parenting. Both forms of violence increase children's risk for clinically significant disruptive behavior, which can place them on a developmental trajectory associated with serious psychological impairment later in life. Although it is hypothesized that IPV behaviors may spillover into harsh parenting, and thereby influence risk for disruptive behavior, relatively little is known about these processes in families with young children. The current study examines the overlap of the quality and frequency of psychological and physical forms of IPV and harsh parenting, and tests whether harsh parenting mediates the relationship between IPV and child disruptive behavior in a diverse cross-sectional sample of 81 children ages 4 to 6 years. Results suggest that mothers reporting a greater occurrence of psychologically aggressive IPV (e.g., yelling, name-calling) more often engage in psychological and physical aggression toward their children (odds ratios [ORs] = 4.6-9.9). Mothers reporting a greater occurrence of IPV in the form of physical assault more often engage in mild to more severe forms of physical punishment with potential harm to the child (ORs = 3.8-5.0). Psychological and physical forms of IPV and harsh parenting all significantly correlated with maternal reports of child disruptive behavior (r = .29-.40). Psychological harsh parenting partially mediated the association between psychological IPV and child disruptive behavior. However, a significant direct effect of psychological IPV on preschool children's disruptive behavior remained. Implications for child welfare policy and practice and intervention, including the need for increased awareness of the negative impact of psychological IPV on young children, are discussed.


Long-term data on the efficacy and tolerability of lamivudine plus dolutegravir as a switch strategy in a multi-centre cohort of HIV-1-infected, virologically suppressed patients.
Results from clinical trials and observational studies suggest that lamivudine plus dolutegravir (3TC+DTG) could be an effective and tolerated option for simplification in human immunodeficiency virus (HIV)-1-positive patients. This observational study enrolled HIV-1-infected, virologically suppressed patients switching to 3TC+DTG. Kaplan-Meyer survival analysis was performed to evaluate time to virological failure (VF; defined by a single HIV-RNA determination ≥1000 copies/mL or by two consecutive HIV-RNA determinations ≥50 copies/mL) and time to treatment discontinuation (TD; defined as interruption of either 3TC or DTG), Cox regression was performed to assess predictors, and linear mixed model was performed for repeated measures to measure changes in immunological and metabolic parameters. Five hundred and fifty-six patients were eligible for analysis. Their median CD4+ count at baseline was 668 cells/mm3 and median time of virological suppression was 88 months. Estimated probabilities of maintaining virological suppression at 96 and 144 weeks of follow-up were 97.5% [standard deviation (SD) 0.8] and 96.5% (SD 1.0), respectively. Years since HIV diagnosis was the only predictor of VF. In patients with time of virological suppression <88 months, the rate of VF was higher in the presence of the M184V mutation. Estimated probabilities of remaining on 3TC+DTG at 96 and 144 weeks of follow-up were 79.2% (SD 1.9) and 75.2% (SD 2.2), respectively. A significant increase in CD4 cell count (+44 cells/mm3, P=0.015), CD4/CD8 ratio (+0.10, P=0.002) and high-density lipoprotein cholesterol (+5.4 mg/dL, P=0.036) was found at 144 weeks of follow-up; meanwhile, total cholesterol (-9.1 mg/dL, P=0.007) and triglycerides (-2.7, P=0.009) decreased significantly. These findings confirm the efficacy and tolerability of 3TC+DTG in virologically suppressed patients.


Investigation of the Prevalence of Oral Mucositis and Associated Risk Factors in Chronic Obstructive Pulmonary Disease Patients: A Prospective Cross-Sectional Study.
This study was conducted to determine the prevalence of oral mucositis in chronic obstructive pulmonary disease patients and the risk factors likely to be effective in the development of oral mucositis. This prospective cross-sectional study was carried out with 147 patients with chronic obstructive pulmonary disease who were monitored for 7 days from their admittance to the chest diseases clinic of a university hospital between January 2021 and January 2022. Patient Information Form, Oral Assessment Guide, and World Health Organization Mucositis Grading Scale were used to obtain the study data. While 81% of the patients in the study were male, the mean age was 64.6 ± 10.0 years, and the duration of chronic obstructive pulmonary disease diagnosis was 8.58 ± 6.52 years. In the study, while 61.9% of the total 147 patients monitored for 7 days had findings in favor of oral mucositis, the severity of oral mucositis and the mean total oral mucosa score of the patients were found to increase significantly in these 7 days of the monitoring period (P <.001). As per the logistic regression findings, the duration of chronic obstructive pulmonary disease diagnosis (95% CI 1.05-1.49; P =.010), the presence of medication use containing corticosteroids (95% CI 6.05-72.17; P <.001), and the amount of oxygen administered to the patient (95% CI 1.07-3.44; P =.029) were found to be significant risk factors associated with the development of oral mucositis. The results indicate that the severity of oral mucositis has a tendency to increase in chronic obstructive pulmonary disease patients from the third day of treatment, and oral care is critically important in patients, who are with a longer diagnosis of chronic obstructive pulmonary disease, who are treated with medications containing corticosteroids and who need high-intensity oxygen therapy.


Differences in prefrontal cortex activity based on difficulty in a working memory task using near-infrared spectroscopy.
The Prefrontal cortex (PFC) has been highly related to executive functions such as working memory (WM). This study assesses the activity of the PFC in performing the Sternberg WM task (ST) with three levels of difficulty (easy, medium and hard) using the near-infrared spectroscopy (fNIRS) technique. Participants were 43 young and healthy right-handed women. Nine WM task blocks were pseudo randomly presented, three for each difficulty task. The results showed that the participant's performance was better in the easy trials than in the medium and hard trials. Performance in the medium trials was also better than in the hard ones. Bonferroni-corrected paired post-hoc t-tests indicated higher oxygenation in medium and hard tasks than in the easy ones for times between 13 and 42 s in the left lateral PFC and in both, medial and lateral, right PFC. Significant differences in Oxygenated hemoglobin (HbO), Total hemoglobin (HbT) and oxygenation (Oxy) changes depending on the Sternberg WM task were found. Unlike previous studies with fNIRS and WM, the current study uses a highly controlled WM task that differentiates between encoding, retention and retrieval phases, comparing different levels of task load.


Mechanism of action of inter-alpha-trypsin inhibitor.
Inter-alpha-trypsin inhibitor (I alpha I) is a unique proteinase inhibitor that can be proteolyzed by the same enzymes that are inhibited, to generate smaller inhibitors. This study examines the reactions of I alpha I with trypsin, chymotrypsin, plasmin, and leukocyte elastase. Complexes of I alpha I and proteinase were demonstrated by gel filtration chromatography. Complete digestion of I alpha I by each proteinase was not accompanied by a comparable loss of inhibition of that enzyme or a different enzyme. Following proteolysis, inhibitory activity was identified in I alpha I fragments of molecular weight 50,000-100,000 and less than 40,000. Addition of a second proteinase inhibitor prevented proteolysis. Both I alpha I and its complex with proteinase were susceptible to degradation. Kinetic parameters for both the inhibition and proteolysis reactions of I alpha I with four proteinases were measured under physiological conditions. On the basis of these results, a model for the mechanism of action of I alpha I is proposed: Proteinase can react with either of two independent sites on I alpha I to form an inhibitory complex or a complex that leads to proteolysis. Both reactions occur simultaneously, but the inhibitory capacity of I alpha I is not significantly affected by proteolysis since the product of proteolysis is also an inhibitor. For a given proteinase, the inhibition equilibrium constant and the Michaelis constant for proteolysis describe the relative stability of the inhibition and proteolysis complexes; the second-order rate constants for inhibition and proteolysis indicate the likelihood of either reaction. The incidence of inhibition or proteolysis reactions involving I alpha I in vivo cannot be assessed without knowledge of the exact concentrations of inhibitor and proteinases; however, analysis of inhibition rate constants suggests that I alpha I might be involved in plasmin inhibition.


Using Mouse and Drosophila Models to Investigate the Mechanistic Links between Diet, Obesity, Type II Diabetes, and Cancer.
Many of the links between diet and cancer are controversial and over simplified. To date, human epidemiological studies consistently reveal that patients who suffer diet-related obesity and/or type II diabetes have an increased risk of cancer, suffer more aggressive cancers, and respond poorly to current therapies. However, the underlying molecular mechanisms that increase cancer risk and decrease the response to cancer therapies in these patients remain largely unknown. Here, we review studies in mouse cancer models in which either dietary or genetic manipulation has been used to model obesity and/or type II diabetes. These studies demonstrate an emerging role for the conserved insulin and insulin-like growth factor signaling pathways as links between diet and cancer progression. However, these models are time consuming to develop and expensive to maintain. As the world faces an epidemic of obesity and type II diabetes we argue that the development of novel animal models is urgently required. We make the case for Drosophila as providing an unparalleled opportunity to combine dietary manipulation with models of human metabolic disease and cancer. Thus, combining diet and cancer models in Drosophila can rapidly and significantly advance our understanding of the conserved molecular mechanisms that link diet and diet-related metabolic disorders to poor cancer patient prognosis.


Development and initial examination of a brief intervention for heightened anxiety sensitivity among heroin users.
Anxiety sensitivity (AS) recently has been identified as a potential cognitive vulnerability underlying substance use problems, with some evidence specifically indicating its relevance to heroin. Focusing on the potential utility of interventions centered on increasing willingness to have anxiety-related sensations reduce vulnerability for relapse following substance use treatment, the current article describes the development of a brief (6 session) behavioral treatment for heightened AS among heroin users. The treatment consists of the following components: (a) psychoeducation about anxiety; (b) interoceptive exposure exercises; and (c) skills-training focused on heightening emotional acceptance, tolerance, and nonevaluative awareness (to facilitate willingness). Preliminary data on this treatment are provided in the form of a case study with a 46-year-old African American man in an inner-city residential substance use treatment facility. Results indicate reductions in AS (especially physical concerns), as well as corresponding decreases in heroin cravings and improvements in emotion regulation.


Multiple-probe thermography for estimating the postmortem interval: I. Continuous monitoring and data analysis of brain, liver, rectal and environmental temperatures in 117 forensic cases.
One hundred seventeen forensic postmortem cases have been studied under controlled conditions. In each case, temperatures of the brain, liver, rectum, and the environment were monitored over a period beginning shortly after death and ending up to 60 h postmortem. The four temperature measurements were recorded every 5 to 10 min using the Microwave Thermography System. Rectal and environmental temperatures were measured by electrical thermocouples while brain and liver temperatures were measured using microwave probes. Data acquisition, analogue-to-digital conversion (ADC), and data processing were provided by a microcomputer. The ADC technique is described and its problems are discussed. The data were then transferred to a mainframe computer for extensive curve-fitting and statistical analysis. The microcomputer-based ADC and data logging and acquisition were found to be accurate, fast, easy to implement, and useful for the field. The postmortem rate of human body cooling was found to be adequately represented by triple-exponential equations.


No Clinically Important Difference in Pain Scores After THA Between Periarticular Analgesic Injection and Placebo: A Randomized Trial.
Periarticular analgesic injection (PAI) is being used more commonly for pain relief after orthopaedic surgeries. However, there is conflicting evidence regarding the effectiveness of PAI for post-THA pain relief. In a double-blind, randomized, controlled trial among patients undergoing same-day bilateral THA, with each patient serving as his or her own control, we asked: (1) Did the pain score as measured on a 100-mm VAS differ between the hips that received PAI versus placebo? (2) Were there differences in complications between the treatment and control hips in these patients? Over a 1-year period at one center, 45 patients underwent same-day bilateral THA; three were excluded for prespecified reasons, and two declined participation in this randomized, controlled trial, leaving 40 patients (80 THAs) in the study. Patients randomly received PAI in one hip and placebo in the contralateral hip; patients, surgeons, and nurses were blinded in terms of which hip received the PAI and which hip received a placebo saline injection. The PAI solution included ropivacaine, morphine hydrochloride hydrate, methylprednisolone, ketoprofen, and epinephrine. The primary outcome was the VAS for pain at rest 24 hours after THA, measured using a 100-mm horizontal VAS. The VAS score was compared between two groups and assessed to reach the reported threshold values for the minimum clinically important difference (MCID) of 20 mm for the postoperative VAS score. No patients were lost to followup, and there were no missing data for the primary outcome. Complications that occurred during the trial were recorded prospectively with emphasis on infection, wound complications, nerve palsy and allergic reactions to the injections. There were no clinically important differences between hips treated with the PAI and those treated with the placebo injection at any point. The hips that received PAI had less pain than those receiving placebo 24 hours after THA (16 ± 17 mm versus 22 ± 20 mm; mean difference, 6 mm; 95% confidence interval [CI], 2-9 mm; p = 0.006), but this effect size was below the MCID of 20 mm and thus is unlikely to be clinically important. The hips that received PAI also had better VAS scores in the recovery room (38 ± 29 mm versus 52 ± 33 mm; mean difference 14 mm; 95% CI, 5-23 mm; p = 0.004) and 3 hours after THA than placebo controls (28 ± 22 mm versus 37 ± 24 mm; mean difference 9 mm; 95% CI, 2-16 mm; p = 0.010). Neither of these differences exceeded the MCID and likewise were unlikely to be clinically important. No complications, including surgical site infections, were observed in either group. Periarticular analgesic injection for pain control after THA did not result in a clinically important reduction in pain at any point examined. Given the expense associated with this PAI mixture and the lack of effectiveness outside this timeframe, we cannot recommend its use. Other mixtures or concentrations of drugs may be helpful in short-stay admissions for THA, but this will require further research. Level I, therapeutic study.


Cognitive Outcomes and Psychiatric Symptoms of Retinopathy-Positive Cerebral Malaria: Cohort Description and Baseline Results.
Cerebral malaria (CM) is a common cause of death and disability among children in sub-Saharan Africa. Many prior studies of neuropsychiatric morbidity have been limited by a cross-sectional design or a short duration of follow-up. Most have included subjects who may have presented with coma due to a disease process other than CM. No studies have assessed the relationship between magnetic resonance imaging (MRI) findings and long-term outcomes. The Cognitive Outcomes and Psychiatric symptoms of retinopathy-positive CM (COPS) cohort is the first large (N = 221) prospectively recruited cohort of stringently defined cases of CM and hospital-based, age-matched, non-CM controls in whom cognitive and psychiatric outcomes are assessed with standardized measures semi-annually for up to 5 years. We report baseline characteristics of the cohort and outcomes at 1 month. At enrollment, CM cases were more likely to come from families with fewer socioeconomic resources and to have health characteristics that increase risk for malaria. In children younger than 5 years, cases were delayed in motor, language, and social development by approximately 6 months, compared with controls. More significant delays occurred in those with MRI abnormalities at the 1-month follow-up visit. There were no differences between cases and controls in inhibitory self-control, nor in cognitive function in children ≥ 5 years of age. The latter finding may be related to the smaller sample size, case-control imbalance in socioeconomic status, or the use of cognitive and behavioral assessments that are less culturally appropriate to this population. Continued follow-up will help determine predictors of long-term outcomes.


Total concentration analysis of polycylic aromatic hydrocarbons in aqueous samples with high suspended particulate matter content.
The European water framework directive (WFD) requires priority pollutants to be measured in the whole water sample and not only in the dissolved phase. However, it does not give clear definitions on how to achieve this. To overcome this limitation, a new methodology of sample preparation procedure for the analysis of polycyclic aromatic hydrocarbons on the basis of extraction disks is introduced here. The automatable procedure includes a "one-step" extraction of the analytes both dissolved in the liquid phase of the sample and sorbed to suspended particulate matter. The latter is extracted concurrently with the solid-phase extraction (SPE) material within the elution step of the procedure. Separation, identification, and quantification of the analytes is performed by GC-MS. Results from surface water samples spiked with certified sediment up to 1000 mg/l are presented in this work and compared with results derived from liquid-liquid extraction (LLE). Most measured values are within or at least near certified uncertainty limits of the sediment. The SPE disk method shows much higher recoveries and better precision (relative standard deviations between 2% and 11%) than the standard LLE method. For all substances under investigation, the limits of quantification achieved range between 0.001 and 0.005 microg/l.


Carotid intima-media thickness, hypertension, and dyslipidemia in obese adolescents.
Obesity is a global health problem with growing prevalence in developing countries. Obesity causes chronic inflammation due to imbalances between pro- and anti-inflammatory cytokines. This causes metabolic complications such as dyslipidemia, hypertension, and cardiovascular disorder. Carotid intima-media thickness (CIMT) is a predictor of atherosclerosis which could be measured easily and non-invasively. Early detection of cardiovascular diseases in obese adolescents at risk is hoped to improve outcomes. This is a cross-sectional study on obese adolescents aged 13-16 year old at Pediatric Clinic of Dr. Soetomo General Hospital. Obesity is defined as Body mass index higher than 95th percentiles according to CDC (2000). Dyslipidemia is diagnosed when either an increase in cholesterol, LDL, triglyceride or a decrease in HDL level is found, as recommended by NCPE and American Academy of Pediatrics. Hypertension is defined as an increase of blood pressure > P95 according to age and gender. The differences of CIMT based on dyslipidemia, hypertension, and gender were analyzed with Wilcoxon Mann Whitney with significant p value (p < 0,005). This study included 59 obese adolescents, consisting of 32 (54.2%) male adolescents and 35 (59.3%) female adolescents. Dyslipidemia was found on 38 (64.4%) adolescents and hypertension was found on 35 (59.3%) adolescents. No difference of CIMT was found between obese adolescents with and without dyslipidemia and with and without hypertension based on gender (p > 0.05). No difference of CIMT based on gender between adolescents aged below 18. The high number of dyslipidemia and hypertension in obese adolescents need an early detection of cardiovascular complication.


[Characteristics of asparaginase from Escherichia coli at different stages of purification].
Asparaginase of Escherichia coli obtained according to the previously developed scheme (extraction, acidification, heating with ammonium sulphate, twice repeated precipitation with ethanol and DEAE-cellulose chromatography) was examined by the methods of polyacrylamide gel electrophoresis, isoelectric focusing and molecular weight assay. Asparaginase activity was measured by the Nessler method. With respect to the above characteristics, the asparaginase preparation was very similar to those described in the literature, e.g. Krasnitin (FRG) and Leunase (Japan).