Finding the right balance: a personal journey from individual proteins to membrane-embedded motors: based on a lecture delivered at the 36th FEBS Congress in Torino, Italy, June 2011.
It is now more than 20 years since the prophetic words of John Fenn, announcing his discovery, stated that 'Electrospray spectra have been obtained for biopolymers including oligonucleotides and proteins, the latter having molecular weights up to 130 000, with as yet no evidence of an upper limit' (Fenn JB, Mann M, Kai Meng C, Fu Wong S & Whitehouse CM (1989) Science246, 64-71). Today, with the mass spectra of intact ribosomes at 2.3 MDa becoming almost routine and the first electrospray spectra of membrane-embedded motors being recorded recently, new challenges are emerging. Knowledge of the intact mass of a protein or complex is only part of the MS information available. Data from the disruption of protein complexes in solution and gas phases are leading to subunit interaction maps and architectural models. Such models are enhanced by coupling with ion mobility in which the collision cross-section of a protein complex can be defined. Linking these attributes with knowledge of subunit dynamics and the role of post-translational modifications on the stability and interactions within complexes is increasing our understanding of the factors that stabilize and convert protein complexes between different quaternary states. From our earliest experiments, studying the folding of individual proteins, through to the characterization of membrane-embedded motors, it is clear that the full potential of electrospray in structural biology has yet to be realized. The present review offers a personal view of the transition from determining the mass of an individual protein to elucidating the structure and dynamics of heterogeneous assemblies in the megadalton mass range.


Concurrent use of cigarettes and smokeless tobacco among US males and females.
The current study describes concurrent use of cigarettes and smokeless tobacco (CiST) among males and females and evaluates factors associated with CiST use. Cross-sectional data were drawn from the 2010 Behavioral Risk Factor Surveillance System (BRFSS). Weighted stratified analyses were performed to find associations between CiST use and sociodemographic factors by gender. CiST users were compared to three different tobacco use groups: nonusers, exclusive smokers, and exclusive ST users. Younger age and heavy alcohol consumption were consistently associated with increased odds of CiST use among both males and females, and regardless of comparison group. Among males, education was inversely related to CiST use, and these findings were consistent in all three comparisons. Among women, those unable to work or out of work were more likely to be CiST users, which was consistent across comparisons. American Indian females had higher odds of CiST use than White females when nontobacco users or smokers were the comparison group. This study identified sociodemographic characteristics associated with CiST use, and differences in these associations among women and men. Additionally, this study highlights the need to carefully consider what comparison groups should be used to examine factors associated with CiST use.


Incomplete Ionization of a 110 meV Unintentional Donor in β-Ga2O3 and its Effect on Power Devices.
Understanding the origin of unintentional doping in Ga2O3 is key to increasing breakdown voltages of Ga2O3 based power devices. Therefore, transport and capacitance spectroscopy studies have been performed to better understand the origin of unintentional doping in Ga2O3. Previously unobserved unintentional donors in commercially available [Formula: see text] Ga2O3 substrates have been electrically characterized via temperature dependent Hall effect measurements up to 1000 K and found to have a donor energy of 110 meV. The existence of the unintentional donor is confirmed by temperature dependent admittance spectroscopy, with an activation energy of 131 meV determined via that technique, in agreement with Hall effect measurements. With the concentration of this donor determined to be in the mid to high 1016 cm-3 range, elimination of this donor from the drift layer of Ga2O3 power electronics devices will be key to pushing the limits of device performance. Indeed, analytical assessment of the specific on-resistance (Ronsp) and breakdown voltage of Schottky diodes containing the 110 meV donor indicates that incomplete ionization increases Ronsp and decreases breakdown voltage as compared to Ga2O3 Schottky diodes containing only the shallow donor. The reduced performance due to incomplete ionization occurs in addition to the usual tradeoff between Ronsp and breakdown voltage.


In vitro simultaneous mapping of the partial pressure of oxygen, pH and inorganic phosphate using electron paramagnetic resonance.
The partial pressure of oxygen (pO2) and the extracellular pH in the tumour microenvironment are essential parameters for understanding the physiological state of a solid tumour. Also, phosphate-containing metabolites are involved in energy metabolism, and interstitial inorganic phosphate (Pi) is an informative marker for tumour growth. This article describes the simultaneous mapping of pO2, pH and Pi using 750 MHz continuous-wave (CW) electron paramagnetic resonance (EPR) and a multifunctional probe, monophosphonated trityl radical p1TAM-D. The concept was demonstrated by acquiring three-dimensional (3D) maps of pO2, pH and Pi for multiple solution samples. This was made possible by combining a multifunctional radical probe, fast CW-EPR spectral acquisition, four-dimensional (4D) spectral-spatial image reconstruction, and spectral fitting. The experimental results of mapping pO2, pH and Pi suggest that the concept of simultaneous mapping using EPR is potentially applicable for the multifunctional measurements of a mouse tumour model.


Integrative multiomics analysis identifies a metastasis-related gene signature and the potential oncogenic role of EZR in breast cancer.
Distant metastasis is a major cause of increased mortality in breast cancer patients, but the mechanisms underlying breast cancer metastasis remain poorly understood. In this study, we aimed to identify a metastasis-related gene (MRG) signature for predicting progression in breast cancer. By screening using three regression analysis methods, a 9-gene signature (NOTCH1, PTP4A3, MMP13, MACC1, EZR, NEDD9, PIK3CA, F2RL1 and CCR7) was constructed based on an MRG set in the BRCA cohort from TCGA. This signature exhibited strong robustness, and its generalizability was verified in the Metabric and GEO cohorts. Of the nine MRGs, EZR is an oncogenic gene with a well-documented role in cell adhesion and cell migration, but it has rarely been investigated in breast cancer. Based on a search of different databases, EZR was found to be significantly more highly expressed in both breast cancer cells and breast cancer tissue. EZR knockdown significantly inhibited cell proliferation, invasion, chemoresistance and EMT in breast cancer. Mechanistically, RhoA activation assays confirmed that EZR knockdown inhibited the activity of RhoA, Rac1 and Cdc42. In summary, we identified a nine-MRG signature that can be used as an efficient prognostic indicator for breast cancer patients, and owing to its involvement in regulating breast cancer metastasis, EZR might serve as a therapeutic target.


A case of hyperzincemia with functional zinc depletion: a new disorder?
We report the case of an 1l-y-old boy with a plasma Zn concentration greater than 200 micromol/L, but with symptoms consistent with Zn deficiency. He has had hepatosplenomegaly, rashes, stunted growth (<3rd centile), anemia, and impaired immune function since infancy. He also has vasculitis and osteoporosis. A plasma Zn-binding protein has been separated and characterized by a combination of size exclusion and ion exchange chromatography and electrophoretic studies and by immunologic methods. Antibodies to the partially purified protein have been raised in rabbits. Size exclusion chromatography shows that Zn is bound to a protein with a mass 110000-300000 kD. Electrophoretic and mass spectrometry studies suggest that the protein may be composed of several subunits. One component of the isolated protein reacts with antiserum to alpha2-macroglobulin; immunoprecipitation studies confirm that the protein is not alpha2-macroglobulin or a histidine-rich glycoprotein. Kinetic studies of zinc metabolism in the patient and his mother with stable Zn isotopes show the presence of increased exchangeable Zn, with a rapid flux from plasma to a stable pool. Liver and muscle Zn and Cu concentrations are raised, but with no abnormal liver histology. Immunoreactive metallothionein in the liver is increased. We suggest that this boy may suffer from a previously unrecognized inborn error of Zn metabolism causing symptomatic zinc deficiency.


Underlying mechanisms in atherosclerosis.
A brief resume of advances in atherosclerosis research over the past 30 years is presented, emphasising the multidisciplinary origins of those advances. A somewhat more detailed review of progress in the lipoprotein field is presented as an example illustrative of the pace of research related to atherosclerosis. Finally, an attempt is made to identify some of the research areas that promise to yield valuable new insights in the next 30 years.


Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle.
A major constraint for the effective control and management of helminth parasites is the lack of rapid, high-throughput, routine diagnostic tests to assess the health status of individual animals and herds and to identify the parasite species responsible for these helminthoses. The capability of a multiplex platform for the simultaneous detection of three pasture associated parasite species was evaluated and compared to existing ELISAs. The recombinant antigens 14.2 kDa ES protein for Cooperia oncophora, major sperm protein for Dictyocaulus viviparus and Cathepsin L1 for Fasciola hepatica were recombinantly expressed either in Escherichia coli or Pichia pastoris. Antigens were covalently coupled onto magnetic beads. Optimal concentrations for coupling were determined following the examination of serum samples collected from experimentally mono-infected animals, before and after their infection with the target species. Absence of cross-reactivity was further determined with sera from calves mono-infected with Haemonchus contortus, Ostertagia ostertagi and Trichostrongylus colubriformis. Examination of negative serum samples was characterised by low median fluorescence intensity (MFI). Establishment of the optimal serum dilution of 1:200 was achieved for all three bead sets. Receiver Operating Characteristic analyses were performed to obtain cut-off MFI values for each parasite separately. Sensitivity and specificity at the chosen cut-off values were close to, or 100% for all bead sets. Examination of serum samples collected on different days post infection from different animals showed a high reproducibility of the assays. Serum samples were additionally examined with two already established ELISAs, an in-house ELISA using the recombinant MSP as an antigen and a DRG ELISA using Cathepsin L1 for liver fluke. The results between the assays were compared and kappa tests revealed an overall good agreement. A versatile bead-based assay using fluorescence detection (xMAP technology) was developed to simultaneously detect antibodies against C. oncophora, D. viviparus and F. hepatica in cattle serum samples. This platform provides rapid, high-throughput results and is highly sensitive and specific in comparison to existing serological as well as coproscopical diagnostic techniques.


Termite Attractant from Fungus-Infected Wood.
Field observations suggested that subterranean termites might follow a concentration gradient of attractive material to find decaying wood. Laboratory cultures of the brown rot fungus, Lenzites trabea Pers. ex. Fr., on pine blocks formed a material attractive to the eastern subterranean termites, Reticulitermes flavipes (Kol.) and R. virginicus Banks, and a Costa Rican termite, Nasutitermes columbicus (Holmgren). Such a potent termite attractant may be useful in termite surveys and control.


Study of Healthcare Personnel with Influenza and other Respiratory Viruses in Israel (SHIRI): study protocol.
The Study of Healthcare Personnel with Influenza and other Respiratory Viruses in Israel (SHIRI) prospectively follows a cohort of healthcare personnel (HCP) in two hospitals in Israel. SHIRI will describe the frequency of influenza virus infections among HCP, identify predictors of vaccine acceptance, examine how repeated influenza vaccination may modify immunogenicity, and evaluate influenza vaccine effectiveness in preventing influenza illness and missed work. Cohort enrollment began in October, 2016; a second year of the study and a second wave of cohort enrollment began in June 2017. The study will run for at least 3 years and will follow approximately 2000 HCP (who are both employees and members of Clalit Health Services [CHS]) with routine direct patient contact. Eligible HCP are recruited using a stratified sampling strategy. After informed consent, participants complete a brief enrollment survey with questions about occupational responsibilities and knowledge, attitudes, and practices about influenza vaccines. Blood samples are collected at enrollment and at the end of influenza season; HCP who choose to be vaccinated contribute additional blood one month after vaccination. During the influenza season, participants receive twice-weekly short message service (SMS) messages asking them if they have acute respiratory illness or febrile illness (ARFI) symptoms. Ill participants receive follow-up SMS messages to confirm illness symptoms and duration and are asked to self-collect a nasal swab. Information on socio-economic characteristics, current and past medical conditions, medical care utilization and vaccination history is extracted from the CHS database. Information about missed work due to illness is obtained by self-report and from employee records. Respiratory specimens from self-collected nasal swabs are tested for influenza A and B viruses, respiratory syncytial virus, human metapneumovirus, and coronaviruses using validated multiplex quantitative real-time reverse transcription polymerase chain reaction assays. The hemagglutination inhibition assay will be used to detect the presence of neutralizing influenza antibodies in serum. SHIRI will expand our knowledge of the burden of respiratory viral infections among HCP and the effectiveness of current and repeated annual influenza vaccination in preventing influenza illness, medical utilization, and missed workdays among HCP who are in direct contact with patients. NCT03331991 . Registered on November 6, 2017.


[Enzymatic hydrolysis of willow treated with a steam burst without preliminary water extraction with a high concentration of substrate].
A laboratory reactor equipped with a screw press was used for hydrolysis of steam-SO2 exploded willow Salix caprea by a composition of Trichoderma reesei and Aspergillus foetidus enzyme preparations at high substrate concentrations. Optimal conditions providing the maximal volume of hydrolysis syrup with maximal sugar concentrations were determined. Two different hydrolysis procedures were developed in order to exclude initial washing of steam-pretreated plant raw material by large volumes of water, which is necessary to eliminate the inhibitory effect of explosion by-products on enzymatic hydrolysis. The first procedure included a one-hour-long enzymatic prehydrolysis of the substrate, then separation of sugar syrup containing 40-60 g/l of glucose, 20-25 g/l of xylose, and up to 10% of disaccharides, as well as up to 35% of the initial enzymatic activity, then addition of a diluted acetate buffer (pH 4.5), and subsequent hydrolysis of the substrate by the adsorbed enzymes leading to the final accumulation of up to 140 g/l glucose and up to 15 g/l xylose. In the second scenario, the exploded willow was initially adjusted by alkali to pH 4.5 and then hydrolyzed directly by added enzymes for 24 hours. This procedure resulted in a nearly total polysaccharide hydrolysis and accumulation of up to 170 g/l glucose and 20 g/l xylose. The reasons of inhibition of enzymatic hydrolysis are discussed.


A Systematic Approach to Evaluate Sudden Unexplained Death in Children.
To evaluate in the Netherlands the national outcomes in providing cause of and insights into sudden and unexplained child deaths among children via the Postmortem Evaluation of Sudden Unexplained Death in Youth (PESUDY) procedure. Children aged 0-18 years in the Netherlands who died suddenly were included in the PESUDY procedure if their death was unexplained and their parents gave consent. The PESUDY procedure consists of pediatric and forensic examination, biochemical, and microbiological tests; radiologic imaging; autopsy; and multidisciplinary discussion. Data on history, modifiable factors, previous symptoms, performed diagnostics, and cause of death were collected between October 2016 and December 2021. In total, 212 cases (median age 11 months, 56% boys, 33% comorbidity) were included. Microbiological, toxicological, and metabolic testing was performed in 93%, 34%, and 32% of cases. In 95% a computed tomography scan or magnetic resonance imaging was done and in 62% an autopsy was performed. The cause of death was explained in 58% of cases and a plausible cause was identified in an additional 13%. Most children died from infectious diseases. Noninfectious cardiac causes were the second leading cause of death found. Modifiable factors were identified in 24% of non-sudden infant death syndrome/unclassified sudden infant death cases and mostly involved overlooked alarming symptoms. The PESUDY procedure is valuable and effective for determining the cause of death in children with sudden unexplained deaths and for providing answers to grieving parents and involved health care professionals.


Effects of physioball and conventional floor exercises on early phase adaptations in back and abdominal core stability and balance in women.
The purpose of this study was to compare the effects of 5 weeks of physioball core stability and balance exercises with conventional floor exercises in women. The experimental group (n = 15) performed curl-ups and back extensions on the physioball while the control group (n = 15) performed the same exercises on the floor. Baseline and post-training tests included electromyography (EMG) recordings of the rectus abdominus and erector spinae muscles; abdominal, back, and knee strength measurements with the Cybex Norm System; and 2 unilateral stance balance tests. The physioball group was found to have significantly greater mean change in EMG flexion and extension activity (p = 0.04 and p = 0.01, respectively) and greater balance scores (p < 0.01) than the floor exercise group. No significant changes (p > 0.05) were observed for heart rate or Cybex strength measurements. Early adaptations in a short-term core exercise program using the physioball resulted in greater gains in torso balance and EMG neuronal activity in previously untrained women when compared to performing exercises on the floor.


Chronic Kidney Disease as Oxidative Stress- and Inflammatory-Mediated Cardiovascular Disease.
Generating reactive oxygen species (ROS) is necessary for both physiology and pathology. An imbalance between endogenous oxidants and antioxidants causes oxidative stress, contributing to vascular dysfunction. The ROS-induced activation of transcription factors and proinflammatory genes increases inflammation. This phenomenon is of crucial importance in patients with chronic kidney disease (CKD), because atherosclerosis is one of the critical factors of their cardiovascular disease (CVD) and increased mortality. The effect of ROS disrupts the excretory function of each section of the nephron. It prevents the maintenance of intra-systemic homeostasis and leads to the accumulation of metabolic products. Renal regulatory mechanisms, such as tubular glomerular feedback, myogenic reflex in the supplying arteriole, and the renin-angiotensin-aldosterone system, are also affected. It makes it impossible for the kidney to compensate for water-electrolyte and acid-base disturbances, which progress further in the mechanism of positive feedback, leading to a further intensification of oxidative stress. As a result, the progression of CKD is observed, with a spectrum of complications such as malnutrition, calcium phosphate abnormalities, atherosclerosis, and anemia. This review aimed to show the role of oxidative stress and inflammation in renal impairment, with a particular emphasis on its influence on the most common disturbances that accompany CKD.


Inverted duplication-transposition event in mammalian cells at an illegitimate recombination join.
Illegitimate recombination events in mammalian cells often contain extraneous nucleotides or filler DNA at the recombinant joins. The polyomavirus-transformed cell line 7axB has previously been found to contain 37 base pairs (bp) of filler DNA at one virus-host join of the single insert of integrated viral DNA (A. Hayday, H. E. Ruley, and M. Fried, J. Virol. 44:67-77, 1982). By using a synthetic oligomer of these 37 bp as a probe, we demonstrated that this filler DNA is an inverted duplication of a single-copy rat sequence found 650 bp upstream from this virus-host join. The other virus-host join appears to be the result of a simple illegitimate recombination event between viral and host sequences. This is the first identification of filler DNA as a transposed copy of a chromosomal sequence. The relevance of the recombination events studied to cellular rearrangements and viral integration is discussed.


Medical Students' Response to the COVID-19 Pandemic: Experience and Recommendations from Five Countries.
The COVID-19 pandemic has provided medical students around the globe with unique challenges and opportunities. With formal medical school education and training interrupted, medical students sought innovative ways to contribute to their health-care systems and communities. Their responses could be organized into three categories: clinical (remote clinical care and triage, helping in COVID testing or treatment centers, and contact tracing), nonclinical (PPE acquisition, COVID-related policy and research, and supporting vulnerable groups in the community), and educational (creating materials to educate peers, the community, or community health workers). We present examples of responses developed by students from five countries: Brazil, Nepal, the Philippines, Rwanda, and the United States. We discuss the challenges, outcomes, and recommendations for each case. One critical opportunity for growth is strengthening international collaborations. We hope that these examples provide a framework for medical students to plan coordinated and effective responses to the next pandemic, and further medical student engagement in international collaboration.


Protective effect of Tualang honey against cadmium-induced morphological abnormalities and oxidative stress in the ovary of rats.
To investigate the protective effects of Tualang honey against the toxicity effects induced by cadmium (Cd) on the ovary. A total of 32 female Sprague Dawley rats were taken and randomly divided into four groups (n = 8). Throughout the experimental period of 6 weeks, negative control-NC (vehicle deionized water), positive control-CD (Cd at 5 mg/kg), Tualang honey followed by Cd exposure-TH (Tualang honey at 200 mg/kg and Cd at 5 mg/kg) and Tualang honey control-THC (Tualang honey at 200 mg/kg) groups, were administered orally on a daily basis. Rats exposed to Cd were significantly higher in ovarian weight, number of antral and atretic follicles as compared to the NC group. The disruptive effects of Cd on ovarian follicles were associated with a disruption in gonadotropin hormones and decreases in follicular stimulating hormone (FSH) and luteinizing hormone (LH). Moreover, a significant formation of oxidative stress in ovarian Cd-exposed rats has been proven by increasing the level of lipid peroxidation products (malondialdehyde) and decreasing the levels of enzymatic antioxidant (catalase). Interestingly, a daily supplementation of high antioxidant agents such as Tualang honey in these animals, caused significant improvements in the histological changes. Additionally, less atretic follicles were observed, restoring the normal level of LH and FSH (P < 0.001), and normalizing the ovarian malondialdehyde (P < 0.05) and catalase levels in comparison with CD group (P < 0.05). Tualang honey has protective effects against Cd-induced ovarian toxicity by reducing morphological abnormalities, restoring the normal levels of gonadotropin hormones and stabilizing equilibrium levels of lipid peroxidation and antioxidant enzyme in ovaries of rats.


Simultaneous echocardiographic phonocardiographic recordings at rest and during amyl nitrite administration in patients with mitral valve prolapse.
Simultaneous electrocardiograms, phonocardiograms, and echocardiograms were recorded in 21 patients with mitral valve prolapse. Four patients with holosystolic mitral valve prolapse on echocardiogram had smaller resting end-diastolic volumes than the remaining 17 patients with late systolic echocardiopraphy prolapse (p greater than 0.01). Thirteen of the 17 patients with late systolic prolapse had phonocardiograhically recorded auscultatory phenomena. The initial vibrations of the auscultatory phenomena occurred after the onset of echocardiographic prolapse, but prior to maximal echocardiographic mitral valve prolapse. Amyl nitrite was administered to all patients. Three of the 17 patients with late systolic prolapse developed holosystolic prolapse, while the remaining 14 retained the late systolic prolapse pattern during amyl nitrite inhalation. In these 14 patients, the onset of mitral prolapse occurred earlier in systole due to decrease in the duration of systole prior to onset of mitral valve prolapse (p greater than 0.001). This corresponded with the occurrence of auscultatory phenomena earlier in systole. Twelve patients had left ventricular volumes recorded during amyl nitrite inhalation and all showed a decrease in left ventricular volumes (greater than) 0.001). These findings confirm the temporal relationship of mitral valve prolapse and onset of auscultatory phenomena in these patients. It suggests that the movement of auscultatory phenomena earlier in systole during amyl nitrite inhalation is related to earlier prolapse of the mitral valve, and that a decrease in ventricular valume is a tenable explanation for the earlier onset of prolapse.


Influence of rabbit sire genetic origin, season of birth and parity order on doe and litter performance in an organic production system.
The aim of this study was to compare both the performance of litters derived from two sire genetic origins (SGO), Vienna Blue (VB) and Burgundy Fawn (BF), along successive seasons of birth (SB; winter, spring, summer and autumn), and doe reproductive performance in an organic production system. A total of fifty-eight does consisting of a mixture of crosses of several medium-large size breeds at different parity order (P, 1 = nulliparous; 2 = primiparous; ≥3 = multiparous) and twelve males (6 VB and 6 BF) were housed indoors at environmental conditions that followed seasonality. An extensive reproductive rhythm was used and kits were weaned at 46±6 d of age. Doe reproductive performance and the data of 105 litters (55 from VB and 50 from BF SGO) were recorded throughout the SB. No statistically significant differences related to SGO effect were observed. As regards parity order, multiparous does showed higher live weights (LW) (p<0.05), total born (p<0.01), total born alive (p<0.05) per delivery, and litter weight of born alive (p<0.05), but lower milk output at 21st d than primiparous does (p<0.05). The extensive reproductive rhythm mainly increased litter performance at birth in multiparous does but was not sufficient to permit a complete recovery of body reserves lost during lactation. Autumn SB negatively affected doe LW variation between deliveries. The number of pups born and born alive per delivery (p<0.05) and litter size at 21 d of age and at weaning (p<0.01) were lower during hot SB. Due to the lower litter size of pups born in summer and autumn, their individual weight at 21st d of age and daily individual growth rate 0 to 21 d were higher than those of pups born in winter (p<0.001). Litter performance at 21st d of age and individual pup pre-weaning growth rate were poorer for those born in spring than in other seasons due to the harmful effects of increased environmental temperatures. SB affected most of the performance traits of does and young rabbits reared under the organic farming system. The rabbits seemed better suited to organic rearing conditions during winter than in other seasons. The worst results overall were obtained in the spring SB, whereas the hot SB negatively affected both doe energy balance and prolificacy. In conclusion, the pups of the 2 SGO showed good pre-weaning performance and seemed suited to the organic rabbit production system.


Insight into skin cell-based osteogenesis: a review.
For decades, researchers have been fascinated by the strategy of using cell therapy for bone defects; some progress in the field has been made. Owing to its ample supply and easy access, skin, the largest organ in the body, has gained attention as a potential source of stem cells. Despite extensive applications in skin and nerve regeneration, an increasing number of reports indicate its potential use in bone tissue engineering and regeneration. Unfortunately, few review articles are available to outline current research efforts in skin-based osteogenesis. This review first summarizes the latest findings on stem cells or progenitors in skin and their niches and then discusses the strategies of skin cell-based osteogenesis. We hope this article elucidates this topic and generates new ideas for future studies.


Classification of myalgic encephalomyelitis/chronic fatigue syndrome by types of fatigue.
Persons with myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) often complain of fatigue states (eg, postexertional malaise, brain fog) that are qualitatively different than normal, daily fatigue. Given the heterogeneous nature of ME/CFS, it is likely that individuals with this illness experience these fatigue types differently in terms of severity and frequency. It is also possible that meaningful subgroups of patients exist that exhibit different patterns of the fatigue experience. The purpose of this study was to investigate whether individuals with ME/CFS can be classified in a meaningful way according to the different types of fatigue they experience. One hundred individuals with ME/CFS participated in the study. Individuals that met inclusion criteria were administered the Multiple Fatigue Types Questionnaire (MFTQ), a 5-factor instrument that distinguishes between different types of fatigue. A cluster analysis was used to classify patients into various clusters based on factor subscale scores. Using a 3-factor solution, individuals were classified according to illness severity (low, moderate, severe) across the different fatigue factors. However, a 5-cluster solution enabled participants with moderate to severe fatigue levels to fall into more differentiated clusters and demonstrate distinct fatigue state patterns. These results suggest that fatigue patterns of individuals with ME/CFS are heterogeneous, and that patients may be classified into meaningful subgroups.


Fluted serum relating to growth of both hepatocyte and hepatocellular carcinoma cells.
This study was undertaken to investigate a humoral substance contributing to both primary hepatocytes and hepatocellular carcinoma (HCC) cells in the primitive form. Peripheral blood from rat, 24 h after partial hepatectomy (PH) or sham-operation, was eluted into 50 fractions using an HPLC, and each fraction was bio-assayed. In vivo relationship between PH and tumor growth was also evaluated. Only one, approximately 100 kDa, of the 50 fractions from PH rat stimulated the growth of both types of cell, and the time-lag between PH and HCC injection also correlated significantly with tumor appearance and growth. Serum in PH rats contains certain humoral stimulator(s) contributing to growth of both hepatocyte and HCC cells.


Could the health decline of prehistoric California indians be related to exposure to polycyclic aromatic hydrocarbons (PAHs) from natural bitumen?
The negative health effects of polycyclic aromatic hydrocarbons (PAHs) are well established for modern human populations but have so far not been studied in prehistoric contexts. PAHs are the main component of fossil bitumen, a naturally occurring material used by past societies such as the Chumash Indians in California as an adhesive, as a waterproofing agent, and for medicinal purposes. The rich archaeological and ethnohistoric record of the coastal Chumash suggests that they were exposed to multiple uptake pathways of bituminous PAHs, including direct contact, fume inhalation, and oral uptake from contaminated water and seafood. We investigated the possibility that PAHs from natural bitumen compromised the health of the prehistoric Chumash Indians in California. Exposure of the ancient Chumash Indians to toxic PAHs appears to have gradually increased across a period of 7,500 years because of an increased use of bitumen in the Chumash technology, together with a dietary shift toward PAH-contaminated marine food. Skeletal analysis indicates a concurrent population health decline that may be related to PAH uptake. However, establishing such a connection is virtually impossible without knowing the actual exposure levels experienced by these populations. Future methodological research may provide techniques for determining PAH levels in ancient skeletal material, which would open new avenues for research on the health of prehistoric populations and on the long-term effects of human PAH exposure.


Immunology of progressive multifocal leukoencephalopathy.
The high prevalence of asymptomatic JC polyomavirus (JCV) infection in the general population indicates coexistence with the human host and efficient immune control in healthy individuals. For unknown reasons, kidney-resident archetypic JCV strains can turn into neurotropic JCV strains which in hereditary or acquired states of immunodeficiency cause opportunistic infection and cytolytic destruction of glial cells or granule cell neurons resulting in progressive multifocal demyelination in the central nervous system (CNS) or cerebellar atrophy, respectively. Immunomodulatory or immunosuppressive therapies with specific monoclonal antibodies including natalizumab, efalizumab, and rituximab have increased the risk of progressive multifocal leukoencephalopathy (PML) among treated patients, highlighting that symptomatic JCV infection of the CNS is associated with disturbances of adaptive immunity affecting B cells, antibodies, and CD4(+) and/or CD8(+) T cells. To date, no specific therapy to overcome PML is available and the only way to eliminate the virus from the CNS is to reconstitute global immune function. However, since the identification of JCV as the causative agent of PML 40 years ago, it is still not fully understood which components of the immune system prevent the development of PML and which immune mechanisms are involved in eliminating the virus from the CNS. This review gives an update about adaptive JCV-specific immune responses.


Effects of Sunday Liquor Sales Legalization on Alcohol Policy Attitudes and Alcohol Purchasing Behavior.
We examined how legalization of Sunday alcohol sales relates to attitudes towards Sunday sales, and how both attitudes and alcohol consumption patterns relate to Sunday alcohol purchasing. A total of 1,384 adults of legal drinking age completed a survey one year post-legalization of Sunday sales. A majority of respondents (51%) were supportive of Sunday sales legalization both before and after legalization. People were more likely to support Sunday sales legalization if they reported binge drinking (PR: 2.19; CI: 1.51 3.18). Following Sunday sales legalization, 59% of participants reported purchasing alcohol in Minnesota on Sunday. Binge drinking (PR: 1.39; CI: 1.27, 1.52) or supporting Sunday sales legalization (PR: 1.85; CI: 1.56, 2.17) were associated with higher likelihood of purchasing alcohol on Sunday. Legalizing Sunday sales may have increased access to alcohol for people with more unhealthy drinking behaviors.


Preoperative low muscle mass and early postoperative outcomes in children undergoing living donor liver transplantation: A retrospective study.
Low skeletal muscle mass may develop in children with end-stage liver disease, affecting postoperative outcomes. We retrospectively investigated whether preoperative low muscle mass was associated with early postoperative outcomes in pediatric patients undergoing living donor liver transplantation (LDLT). Electronic medical records of children (age below 12 y) who underwent LDLT between February 1, 2007, and January 31, 2018, were reviewed. The cross-sectional areas of psoas, quadratus lumborum, and erector spinae muscles at the level of fourth-fifth lumbar intervertebral disks were measured using abdominal CT images, divided by the square of the height and were added to obtain the total skeletal muscle index (TSMI). The patients were divided into two groups according to the median TSMI in the second quintile (1859.1 mm 2 /m 2 ). Complications in the early postoperative period (within 30 d after surgery) classified as Clavien-Dindo grade 3 or higher were considered major complications. Logistic regression analyses were performed to determine the association between preoperative low muscle mass and early postoperative outcomes. In the study population of 123 patients (median age, 14 mo; range, 8-38 mo) who underwent LDLT, 29% and 71% were classified in the low (mean TSMI, 1642.5 ± 187.0 mm 2 /m 2 ) and high (mean TSMI 2188.1 ± 273.5 mm 2 /m 2 ) muscle mass groups, respectively. The rates of major complications, mechanical ventilation >96 hours, intensive care unit stay >14 days, hospital stay >30 days, and in-hospital mortality were not significantly different between the 2 groups. Additionally, adverse outcomes according to pediatric end-stage liver disease scores and sex were not significantly different between the 2 groups. In conclusion, preoperative low muscle mass defined by TSMI was not associated with early postoperative outcomes in pediatric patients undergoing LDLT.


The dual-wave bolus feature in continuous subcutaneous insulin infusion pumps controls prolonged post-prandial hyperglycaemia better than standard bolus in Type 1 diabetes.
The dual-wave bolus delivers a combination of an immediate normal pre-meal insulin bolus (approximately 3 min) followed by an extended (or square-wave) bolus that is evenly delivered over several hr as programmed by the patient. The purpose of this study was to compare post-prandial glycaemic excursions following a high-fat meal after administration of insulin by normal vs dual-wave bolus. During this prospective, cross-over, repeated measures study, subjects with diabetes and treated with insulin pump therapy were evaluated using the continuous glucose monitoring system (CGMS) following three combinations of meal and bolus type. A control meal or a high-fat meal was given in place of the evening meal on three separate occasions and comparisons were made between: a) the control meal with normal insulin bolus delivery, b) the high-fat meal with normal insulin bolus delivery, and c) the high-fat meal with dual-wave insulin bolus delivery. Although mean baseline CGMS values were similar in each of the three combinations of meal and bolus type (p=0.54) and in the three hr immediately following the meal (p=0.64, p=0.83, p=1.0), when compared to the control meal/normal bolus and high-fat meal/dual-wave bolus combinations, CGMS profiles disclosed significantly elevated post-prandial glucose in hr 5 through 14 (p<0.05) following the high-fat/normal bolus combination. Prolonged post-prandial glycaemic excursions are identified using the CGMS. Treating post-prandial hyperglycaemia with dual-wave insulin delivery may help manage chronic hyperglycaemia in patients with diabetes.


Feeding response of Aedes aegypti and Anopheles dirus (Diptera: Culicidae) using out-of-date human blood in a membrane feeding apparatus.
The colonization of Aedes aegypti and Anopheles dirus was performed using out-of-date human blood from a blood bank as a nutritional supply dispensed from a common artificial feeder. Preserved human blood was collected and used for feeding on days 5, 15, and 25 after date of expiration and dispensed from a common artificial feeder to rear the mosquitoes. Ae. aegypti had a feeding rate of 78.7, 62, and 18% at the respective intervals while An. dirus had a rate of 80, 56.8, and 7.3% on the same respective days. Direct feeding on live hamsters resulted in a rate of 96 and 90% for Ae. aegypti and An. dirus, respectively. Although egg production rates decreased from the day 5 feeding to the day 25 feeding, all of the developmental stages resulting from An. dirus fed at day 5 and 15 showed insignificant differences when compared with direct feeding on the blood of a hamster.


Budd-Chiari syndrome due to giant hydatid cyst: a case report and brief literature review.
Budd-Chiari syndrome is an uncommon disorder characterized by the thrombotic or non-thrombotic obstruction of hepatic venous outflow anywhere along the venous course from the hepatic venules to the junction of the inferior vena cava and the right atrium. The etiology of Budd-Chiari syndrome is classified as primary, attributable to intrinsic intraluminal thrombosis or the development of venous webs; or secondary, caused by intraluminal invasion by a parasite or malignant tumor or extraluminal compression by an abscess, solid tumor, or cyst, such as a hydatid cyst. In this study, we present a case of a giant hydatid cyst manifesting Budd-Chiari syndrome symptoms and signs by compressing the inferior vena cava and hepatic veins. In brief, the case demonstrates that hydatid disease should be considered in the differential diagnosis of Budd-Chiari Syndrome in areas such as Turkey, where hydatid disease is endemic.


Family of Polynuclear Nickel Cagelike Phenylsilsesquioxanes; Features of Periodic Networks and Magnetic Properties.
A new family of bi-, tetra-, penta-, and hexanickel cagelike phenylsilsesquioxanes 1-6 was obtained by self-assembly and transmetalation procedures. Their crystal structures were established by single-crystal X-ray analysis, and features of crystal packing relevant to the network formation were studied by a topological analysis. Compounds 1, 2, and 4 are isolated architectures, while 3, 5, and 6 present extended 1D and 3D networks. The investigation of magnetic properties revealed the presence of ferro- (1 and 3-5) or antiferromagnetic (2 and 6) interactions between Ni(II) ions, giving rise in the most cases (1, 2, and 4-6) to the presence of a slow relaxation of the magnetization, which can originate from the spin frustration.


Inhibition of DNA synthesis by cytosine arabinoside: relation to response of acute non-lymphocytic leukemia to remission induction therapy and to stage of the disease.
The sensitivity of leukemic marrow cell DNA synthesis to cytosine arabinoside (araC) exposure in vitro was studied in specimens obtained from patients with acute non-lymphocytic leukemia. Cells from patients who had been treated with araC in the past were more likely to be resistant to the effect of araC on DNA synthesis than cells obtained from patients who had not been so-treated previously. Resistance to the effects of araC on DNA synthesis was associated with the failure of high-dose araC therapy to induce remissions in relapsed patients, whereas remission induction failure in previously untreated patients was not associated with araC resistance.


[Experiences with outpatient laparoscopic cholecystectomy].
To determine the feasibility and desirability of laparoscopic cholecystectomy in day care. Prospective, pilot study. Academic Medical Centre, Amsterdam, the Netherlands. Fifteen patients (ASA I/II according to the American Society of Anesthesiologists) with symptomatic gallstones underwent laparoscopic cholecystectomy with the intention to discharge them the same day. Data were collected about: pain (visual analogue scale); pain medication before and after discharge; (rc)admissions; consultations of general practitioner or day care centre; complications; satisfaction of the patients about this treatment modality. Fourteen patients could be discharged after 6.2 hour (standard deviation 1.2) of observation. The pain scores and medication use were low. Readmissions did not take place and there were no consultations of general practitioners or the day care centre. These patients had no complications. Thirteen of them preferred day care over admission in hospital. One patient was admitted because of colicky pains and a common bile duct stone was diagnosed, which was removed endoscopically. Laparoscopic cholecystectomy in day care appears feasible in patients with symptomatic gallstones and no functional incapacities if day care facilities are present.


Protein requirements of infants and children.
During the last 35 years there have been various published assessments of human protein needs, including those of infants and children. Most recently, the Institute of Medicine of the US National Academies has published its report on Dietary Reference Intakes (DRI) for Macronutrients, and WHO/FAO/UNU have convened a new Expert Consultation, which is due to be published soon. Although there have been a number of published studies on children's protein requirements determined by the nitrogen balance technique, the results of these studies in themselves are insufficient to derive requirement values for all ages. Instead, a meta-analysis of the data from a range of studies in children has been used to derive values for the requirement for maintenance (i.e. no growth), and for the efficiency of utilization of dietary protein for growth. These values were then combined with age-specific rates of protein deposition to calculate the average requirement at any age. New values for protein deposition in children have been derived from studies of potassium- 40 accumulation in infants and children, and these have been employed in new calculations of the average protein requirement. By combining values for the maintenance requirement, derived from adults, with the requirement for growth, estimated by analysis of the data for total-body potassium at different ages, the age-specific average protein requirement was determined. The safe level of intake is the amount of dietary protein that will provide the needs of almost all of the specified age group, and this is usually taken as the average requirement plus 2 times the standard deviation (SD) of the requirement. The SD of the children's requirement was determined by combining the SD for maintenance (from the adult data) with the SD for protein deposition, derived by combining data for rates of growth of body mass and data for the whole body potassium-40 content at different ages. The resulting values for the average protein requirement range from 1.12 g/kg/day at age 6 months to 0.74 g/kg/day at 10 years, with a small decline towards the adult value thereafter. The corresponding values for the safe level are 1.43 g/kg/day at 6 months and 0.91 g/kg/day at 10 years.


Shear-dependent changes in the three-dimensional structure of human von Willebrand factor.
The three-dimensional tertiary structure of human von Willebrand Factor (vWF) on a hydrophobic surface under aqueous conditions and different shear stress regimes was studied by atomic force microscopy (AFM). vWF was imaged by AFM at molecular level resolution under negligible shear stress, under a local applied shear force (7.4 to 19 nN) using the AFM probe in contact mode scanning, and after subjecting vWF to a range of shear stress (0 to 42.4 dyn/cm2) using a rotating disk system. The results demonstrate that vWF undergoes a shear stress-induced conformational transition from a globular state to an extended chain conformation with exposure of intra-molecular globular domains at a critical shear stress of 35 +/- 3.5 dyn/cm2. The globular vWF conformation (149 nm by 77 nm and height 3.8 nm) is representative of native vWF after simple diffusion to the hydrophobic surface, followed by adhesion and some spreading. In a shear stress field above the critical value, protein unfolding occurs and vWF is observed in extended chain conformations oriented in the direction of the shear stress field with molecular lengths ranging from 146 to 774 nm and 3.4 nm mean height. The shear stress-induced structural changes to vWF suggest a close conformation-function relationship in vWF properties for thrombogenesis in regions of high shear stress.


IDDM and autoimmune thyroid disease in the pediatric age group.
Diabetic patients have increased chances of developing autoimmune thyroid disease. Thyroid autoantibodies (Th-AAb) are more frequent in IDDM children than in the general population, ranging between 7 and 40%. As markers of thyroid autoimmunity, we assessed Th-AAb (MsA and TgA) cross-sectionally in 212 children and adolescents (93 girls and 119 boys) aged 1.2-21 years with IDDM from 0-18 years, and longitudinally in 90/212 (43 girls and 47 boys) at diagnosis and during a 3-10 year follow-up. In the cross-sectional study Th-AAb were found in 22/93 girls (23.7%) and 13/119 boys (10.9%). In the longitudinal study Th-AAb were observed at diagnosis in 6 patients, and during the follow-up in 9 girls. In 11/15 Th-AAb positive patients anti-nuclear antibodies were also present. Hormonal assessment revealed hypothyroidism in 3 girls (afterwards on replacement therapy), thyroid ultrasonography showed abnormal patterns in 5 girls, fine needle aspiration biopsy confirmed Hashimoto's thyroiditis in 9 (8 girls and 1 boy), with a higher frequency than that reported among healthy subjects (1-2%). Thyrotoxicosis also occurs with increased frequency in diabetic children than in the general population. We observed Graves' disease in only 1/212 IDDM patients, a 13 year-old boy in whom thyrotoxicosis developed 4 years after diabetes was diagnosed. The high prevalence of thyroid autoimmunity in our patients, particularly in females, suggests that diabetic children and adolescents should be screened for thyroid autoimmunity even if asymptomatic for hypo- or hyperthyroidism. Patients with IDDM and autoimmune thyroid disease should be evaluated for autoantibodies against other organs, such as adrenal glands and gastric mucosa. It is known that patients affected by type 1 (insulin-dependent) diabetes mellitus (IDDM) may have autoantibodies against different organs, such as thyroid, adrenal glands, gastric mucosa, parathyroid, with or without evident dysfunction of the target organ /1-8/. Among organ-specific disorders, autoimmune thyroid disease (ATD) is frequently associated with IDDM and the presence of thyroid autoantibodies (Th-AAb) has been considered a risk factor for the development of hypo- or hyperthyroidism /9/.


[Death versus feelings: a reality in the world of medical students].
The purpose of this investigation was to unveil the feelings of medical students while facing the phenomena of death, and also the meaning of these experiences on their lived-world and its implications to their academic training. The investigation was carried out with 10 students who are regularly registered in the Medical School of the Pontificia Universidade Católica do Rio Grande do Sul (PUCRS), Porto Alegre, Brazil, attending between the 3rd and the 6th year. The qualitative approach, with phenomenological basis, along with theoretical assumptions of Merleau-Ponty and the information analysis method proposed by Giorgi (1988) and Comiotto (1992), disclosed five phenomenological essences: The meanings of death; Conflict between life and death; Feelings experienced on the interfaces of death; Medical training; Personal transformation as a path to professional transformation. The findings of this study evidence the feelings and their implications to the process of personal and professional development of the students as primordial and undeniable phenomenon. Considering this relevance, a psycho-pedagogical proposal is presented to educate feelings.


Nanocrystalline diamond sensor targeted for selective CRP detection: an ATR-FTIR spectroscopy study.
Protein immobilization on functionalized fluorine-terminated nanocrystalline (NCD) films was studied by attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy using an immobilization protocol developed to specifically bind C-reactive protein (CRP). Using an ATR-FTIR spectroscopy method employing a force-controlled anvil-type configuration, three critical steps of the ex situ CRP immobilization were analyzed. First, the NCD surface was passivated by deposition of a copolymer layer consisting of polyethylene oxide and polypropylene oxide. Second, a synthetic modified polypeptide binder with high affinity to CRP was covalently attached to the polymeric film. Third, CRP dissolved in aqueous buffer in concentrations of 10-20 μg/mL was added on the functionalized NCD surface. Both the amide I and II bands, due to the polypeptide binder and CRP, were clearly observed in ATR-FTIR spectra. CRP amide I bands were extracted from difference spectra and yielded bands that agreed well with the reported amide I band of free (non-bonded) CRP in solution. Thus, our results show that CRP retains its secondary structure when it is attached to the polypeptide binders. Compared to previous IR studies of CRP in solution, about 200 times lower concentration was applied in the present study. Graphical Abstract Direct non-destructive ATR-FTIR analysis of C-reactive protein (CRP) selectively bound to functionalized nanocrystalline diamond (NCD) sensor surface.


Short-term muscle power and speed in preschoolers exhibit stronger tracking than physical activity.
The purpose of this study was to examine the tracking of short-term muscle power, speed, and physical activity over a 15-month period in a sample of healthy Canadian preschool-aged children. Seventeen preschoolers (age, 4.4 ± 0.8 years) completed exercise testing and physical activity monitoring at baseline and follow-up separated by 14.6 ± 4.1 months. Short-term muscle power was measured using a modified 10-s Wingate test with peak power and mean power normalized to body mass. Speed was assessed with a 25-m dash. Physical activity was measured by accelerometry (Actigraph GT1M) using a 3-s epoch over 7 consecutive days. Total physical activity and moderate-to-vigorous physical activity, expressed as a percentage of accelerometer wear time, were examined. Tracking of the variables between year 1 and year 2 was analyzed using Spearman rank order correlations and Kappa statistics. Paired t-tests were used to assess differences in performance and physical activity between year 1 and year 2. Total physical activity was not significantly different at year 2 (p > 0.05) and showed fair tracking (r = 0.51, p = 0.05; ĸ = 0.30). Moderate-to-vigorous physical activity was increased at year 2 (p = 0.03) and exhibited poor tracking (r = 0.29, p = 0.28; ĸ = 0.00). Short-term muscle power and speed was increased at year 2 (p < 0.0001) and exhibited significant tracking: peak power (r = 0.72, p = 0.001; ĸ = 0.46), mean power (r = 0.83, p = 0.00004; ĸ = 0.82), and 25-m dash (r = 0.82, p = 0.0001; ĸ = 0.47). Moderate-to-vigorous physical activity increased in this sample of boys and girls during the preschool years, and short-term muscle power and speed exhibited stronger tracking than physical activity.


The effect of induced blur on the Beery-Buktenica developmental test of visual-motor integration and its supplemental tests.
The Beery-Buktenica Test of Visual-Motor Integration (Beery VMI) is a commonly used standardized test of visual-motor integration. Performance on the test is related to academic achievement, but the effect of reduced visual acuity on test results is unknown. This study determined the visual acuity demand and the spacing of the test forms for the Beery VMI and its supplemental tests and investigated the effect of induced optical blur on test results in both adults and children. The overall size and critical detail size of each form and the distance between the center of each form and its adjacent crowding source were measured. The visual acuity demand and angular separation of forms were calculated. Two groups of participants (adults aged ≥18 years [n = 19] and children aged 7-12 years [n = 20]) completed four sessions in which they performed the Beery VMI and the supplemental tests under different blur conditions (habitual near correction, monocular spherical blur, binocular spherical blur and binocular astigmatic blur) in a randomized order. Stroke width, overall form size and box size remained constant throughout the Beery VMI, whereas these reduced with increasing difficulty for the supplemental tests. Reduced near visual acuity from simulated blur resulted in reduced mean scores for the Beery VMI and its supplemental tests, compared with habitual near vision in both adults and children. Binocular spherical blur had the most detrimental effect (p<0.001), followed by binocular astigmatic blur (p<0.001) then monocular spherical blur (p = 0.022). In individuals with uncorrected spherical or astigmatic ametropia, reduced scores on the Beery VMI and its supplemental tests may be due to impaired near visual acuity and not reflect reduced visual-motor abilities. This highlights the importance of excluding reduced near visual acuity as a cause of reduced performance before diagnosing impairment and initiating treatment strategies for visual-motor integration.


Using miscue analysis to assess comprehension in deaf college readers.
For over 30 years, teachers have used miscue analysis as a tool to assess and evaluate the reading abilities of hearing students in elementary and middle schools and to design effective literacy programs. More recently, teachers of deaf and hard-of-hearing students have also reported its usefulness for diagnosing word- and phrase-level reading difficulties and for planning instruction. To our knowledge, miscue analysis has not been used with older, college-age deaf students who might also be having difficulty decoding and understanding text at the word level. The goal of this study was to determine whether such an analysis would be helpful in identifying the source of college students' reading comprehension difficulties. After analyzing the miscues of 10 college-age readers and the results of other comprehension-related tasks, we concluded that comprehension of basic grade school-level passages depended on the ability to recognize and comprehend key words and phrases in these texts. We also concluded that these diagnostic procedures provided useful information about the reading abilities and strategies of each reader that had implications for designing more effective interventions.


Synthesis of α-functionalized ketones by visible-light promoted oxygenation of alkenes.
This article gives a short review on the visible-light promoted synthesis of α-functionalized ketones from alkenes. Various α-functionalized ketones, including α-halo, sulfinyl/sulfonyl, aryl, azido and thiocyanato ketones can be synthesized from the reaction of alkenes with different radical species generated by visible-light catalysis. These transformations typically use dioxygen as the terminal oxidant and do not require a transition metal to activate the double bond, which is quite different from the transition-metal catalysed reactions (Wacker oxidation or hydroformylation).


Determination of the components of analgesic mixtures using high-performance thin-layer chromatography.
A spectrodensitometric method is described for the determination of the components of two analgesic mixtures. For the first mixture (paracetamol-ascorbic acid-caffeine-phenylephrine), the pharmaceutically active components were separated from each other and closely related degradation products and impurities on high-performance thin-layer chromatography (F254) plates using methylene chloride-ethyl acetate-ethanol-formic acid (3.5 + 2 + 4 + 0.5) and methylene chloride-ethyl acetate-ethanol (5 + 5 + 1) as the developing systems. The other mixture (phenazone-phenacetin-caffeine) was separated efficiently from the degradation products using the same plates and acetonitrile-chloroform (1 + 1) as the mobile phase. The proposed method was used to determine these mixtures in commercial tablets.


Ontogenesis of the high affinity NADPH-dependent cytosolic 3,5,3'-triiodo-L-thyronine-binding protein in rat.
Studies were undertaken to define the concentration of the high affinity NADPH-dependent cytosolic 3,5,3'-triiodo-L-thyronine (T3)-binding protein (CTBP) in various tissues of adult rats and in those of developing rats. The maximal binding capacity (MBC) for T3 binding was calculated from the data obtained by Scatchard analysis of T3 binding to the charcoal-extracted cytosol in the presence of 50 microM NADPH. There were no significant differences in the affinity constant among various tissues, whereas the MBC was different among tissues in adult rats. When the levels of MBC were expressed by moles/DNA concentration, the order was as follows; kidney greater than heart greater than or equal to cerebrum greater than or equal to liver much greater than testis greater than cerebellum greater than spleen. The CTBP was not detected before birth in liver, heart, and spleen. The CTBP in these tissues emerged 5 days after the birth, and increased during 6 weeks after the birth. The CTBP in kidney was not detected before birth but was detected at the time of birth. The level of MBC in kidney increased during 3 weeks after the birth. In contrast, the CTBP was detected 5 days before birth, and the level of MBC increased at the time of birth both in cerebrum and cerebellum. The MBC in cerebrum gradually decreased after birth, but began to increase again 2 weeks after the birth. The level of MBC in cerebellum did not increase again. These results suggested that the target tissues of thyroid hormone are enriched with NADPH-dependent CTBP, and that the changes in the concentration of the CTBP are related to the growth of tissues. It was speculated that the CTBP has a specific function in cerebrum at the time of birth although the action of the CTBP is not certain.


Polycomb complexes and the propagation of the methylation mark at the Drosophila ubx gene.
Polycomb group proteins are transcriptional repressors that control many developmental genes. The Polycomb group protein Enhancer of Zeste has been shown in vitro to methylate specifically lysine 27 and lysine 9 of histone H3 but the role of this modification in Polycomb silencing is unknown. We show that H3 trimethylated at lysine 27 is found on the entire Ubx gene silenced by Polycomb. However, Enhancer of Zeste and other Polycomb group proteins stay primarily localized at their response elements, which appear to be the least methylated parts of the silenced gene. Our results suggest that, contrary to the prevailing view, the Polycomb group proteins and methyltransferase complexes are recruited to the Polycomb response elements independently of histone methylation and then loop over to scan the entire region, methylating all accessible nucleosomes. We propose that the Polycomb chromodomain is required for the looping mechanism that spreads methylation over a broad domain, which in turn is required for the stability of the Polycomb group protein complex. Both the spread of methylation from the Polycomb response elements, and the silencing effect can be blocked by the gypsy insulator.


[Effect of anabasine hydrochloride on embryogenesis in white rats and rabbits].
The author reports the results of testing the embryotropic activity of anabasineee hydrochloride in doses of 15, 5, and 3 mg/kg. It was found that oral administration of the drug to pregnant rats in doses exceeding 10-fold (5 mg/kg) and 20-fold (10 mg/kg) the therapeutic ones produced no adverse effect on the fetus. In a dose of 15 mg/kg the drug exerted a moderate embryotoxic action. Multiple oral administration of anabasin hydrochloride to rats in a dose of 3 mg/kg produced no embryotoxic effect. It is concluded that anabasin hydrochloride has no teratogenic properties.


Changes in the timing of departure and arrival of Irish migrant waterbirds.
There have been many recent reports across Europe and North America of a change in the timing of arrival and departure of a range of migrant bird species to their breeding grounds. These studies have focused primarily on passerine birds and climate warming has been found to be one of the main drivers of earlier arrival and departure in spring. In Ireland, rising spring temperature has been shown to result in the earlier arrival of sub-Saharan passerine species and the early departure of the Whooper Swan. In order to investigate changes in spring arrival and departure dates of waterbirds to Ireland, we extracted latest dates as an indicator of the timing of departure of winter visitors (24 species) and earliest dates as an indicator of the timing of arrival of spring/summer migrants (2 species) from BirdWatch Ireland's East Coast Bird reports (1980-2003). Three of the winter visitors showed evidence of later departure and one of earlier departure whereas one of the spring/summer visitors showed evidence of earlier arrival. In order to determine any influence of local temperature on these trends, we analysed data from two synoptic weather stations within the study area and found that spring (average February, March and April) air temperature significantly (P < 0.05) increased at a rate of 0.03 °C per year, which was strongly correlated with changes in latest and earliest records. We also tested the sensitivity of bird departure/arrival to temperature and found that Northern Pintail would leave 10 days earlier in response to a 1 °C increase in spring temperature. In addition, we investigated the impact of a large-scale circulation pattern, the North Atlantic Oscillation (NAO), on the timing of arrival and departure which correlated with both advances and delays in departure and arrival. We conclude that the impact of climate change on earliest and latest records of these birds is, as expected, species specific and that local temperature had less of an influence than large-scale circulation patterns.


A new regulatory function of the region proximal to the RGG box in the fragile X mental retardation protein.
Fragile X mental retardation protein (FMRP) is required for normal cognition. FMRP has two autosomal paralogs, which although similar to FMRP, cannot compensate for the loss of FMRP expression in brain. The arginine- and glycine-rich region of FMRP (the RGG box) is unique; it is the high-affinity RNA-binding motif in FMRP and is encoded by exon 15. Alternative splicing occurs in the 5' end of exon 15, which is predicted to affect the structure of the distally encoded RGG box. Here, we provide evidence that isoform 3, which removes 25 amino acids from the 5' end of exon 15, has an altered conformation that reduces binding of a specific antibody and renders the RGG box unable to efficiently associate with polyribosomes. Isoform 3 is also compromised in its ability to form granules and to associate with a key messenger ribonucleoprotein Yb1 (also known as p50, NSEP1 and YBX1). Significantly, these functions are similarly compromised when the RGG box is absent from FMRP, suggesting an important regulatory role of the N-terminal region encoded by exon 15.


Combined Cardiac and Respiratory Monitoring from a Single Signal: A Case Study Employing the Fantasia Database.
This study proposes a novel method for obtaining the electrocardiogram (ECG) derived respiration (EDR) from a single lead ECG and respiration-derived cardiogram (RDC) from a respiratory stretch sensor. The research aims to reconstruct the respiration waveform, determine the respiration rate from ECG QRS heartbeat complexes data, locate heartbeats, and calculate a heart rate (HR) using the respiration signal. The accuracy of both methods will be evaluated by comparing located QRS complexes and inspiration maxima to reference positions. The findings of this study will ultimately contribute to the development of new, more accurate, and efficient methods for identifying heartbeats in respiratory signals, leading to better diagnosis and management of cardiovascular diseases, particularly during sleep where respiration monitoring is paramount to detect apnoea and other respiratory dysfunctions linked to a decreased life quality and known cause of cardiovascular diseases. Additionally, this work could potentially assist in determining the feasibility of using simple, no-contact wearable devices for obtaining simultaneous cardiology and respiratory data from a single device.


Characterization and repeat analysis of the compact genome of the freshwater pufferfish Tetraodon nigroviridis.
Tetraodon nigroviridis is a freshwater pufferfish 20-30 million years distant from Fugu rubripes. The genome of both tetraodontiforms is compact, mostly because intergenic and intronic sequences are reduced in size compared to other vertebrate genomes. The previously uncharacterized Tetraodon genome is described here together with a detailed analysis of its repeat content and organization. We report the sequencing of 46 megabases of bacterial artificial chromosome (BAC) end sequences, which represents a random DNA sample equivalent to 13% of the genome. The sequence and location of rRNA gene clusters, centromeric and subtelocentric satellite sequences have been determined. Minisatellites and microsatellites have been cataloged and notable differences were observed in comparison with microsatellites from Fugu. The genome contains homologies to all known families of transposable elements, including Ty3-gypsy, Ty1-copia, Line retrotransposons, DNA transposons, and retroviruses, although their overall abundance is <1%. This structural analysis is an important prerequisite to sequencing the Tetraodon genome.


Purification and characterization of thyroid-hormone-binding protein from masu salmon serum. A homolog of higher-vertebrate transthyretin.
We purified a thyroid-hormone-binding protein (THBP) from serum of masu salmon at the stage of smoltification when the concentrations of endogenous thyroid hormones in plasma reach the highest levels. All steps of sequential column chromatography suggest that this THBP is responsible for most L-3,5,3'-triiodothyronine-binding activity in serum at this stage. The molecular mass of this protein was estimated to be 60 kDa by gel filtration but only 15 kDa by SDS/PAGE, which suggests that it is comprised of four identical subunits. The amino acid sequence of its N-terminal portion was highly similar to those of vertebrate transthyretins. These molecular features indicate that masu salmon THBP is a homolog of transthyretins from tetrapods. However, in contrast with mammalian transthyretins, the affinity of masu salmon transthyretin for L-3,5,3'-triiodothyronine was three times greater than for L-thyroxine. This rank order affinity is similar to that of avian and frog transthyretins. Scatchard analysis revealed that masu salmon transthyretin possesses a single class of binding site for L-3,5,3'-triiodothyronine, with a Kd of 13.8 nM at 0 degrees C. Taken together with the data reported by Chang et al. [Eur. J. Biochem. (1999) 259, 534-542], these results suggest that transthyretin has changed from a L-3,5, 3'-triiodothyronine-carrier protein to a L-thyroxine-carrier protein during mammalian evolution.


Pathogenicity and immunogenicity of temperature-sensitive mutants of murine cytomegalovirus.
Two temperature-sensitive (ts) mutants (ts 21 and ts 24), which could complement each other, were isolated from cell culture-passaged murine cytomegalovirus (CC-MCMV). Characterization in vivo revealed that ts 21 lacked pathogenicity for suckling mice, neurovirulence for weanling mice and productive chronic infection in the salivary gland, whereas ts 24 was positive only for the first two parameters; both mutants retained immunogenicity comparable to that of the wild-type CC-MCMV. The immunogenicity of ts 21 and ts 24 was assessed by mortality of the immunized mice after challenge with virulent, salivary gland-passaged MCMV (SG-MCMV) and by replication of the challenge virus in the immunized hosts. The protective immunity conferred by ts 21 was maintained over a period of 3 months without production of infectious viruses in the salivary gland.


A machine learning model of microscopic agglutination test for diagnosis of leptospirosis.
Leptospirosis is a zoonosis caused by the pathogenic bacterium Leptospira. The Microscopic Agglutination Test (MAT) is widely used as the gold standard for diagnosis of leptospirosis. In this method, diluted patient serum is mixed with serotype-determined Leptospires, and the presence or absence of aggregation is determined under a dark-field microscope to calculate the antibody titer. Problems of the current MAT method are 1) a requirement of examining many specimens per sample, and 2) a need of distinguishing contaminants from true aggregates to accurately identify positivity. Therefore, increasing efficiency and accuracy are the key to refine MAT. It is possible to achieve efficiency and standardize accuracy at the same time by automating the decision-making process. In this study, we built an automatic identification algorithm of MAT using a machine learning method to determine agglutination within microscopic images. The machine learned the features from 316 positive and 230 negative MAT images created with sera of Leptospira-infected (positive) and non-infected (negative) hamsters, respectively. In addition to the acquired original images, wavelet-transformed images were also considered as features. We utilized a support vector machine (SVM) as a proposed decision method. We validated the trained SVMs with 210 positive and 154 negative images. When the features were obtained from original or wavelet-transformed images, all negative images were misjudged as positive, and the classification performance was very low with sensitivity of 1 and specificity of 0. In contrast, when the histograms of wavelet coefficients were used as features, the performance was greatly improved with sensitivity of 0.99 and specificity of 0.99. We confirmed that the current algorithm judges the positive or negative of agglutinations in MAT images and gives the further possibility of automatizing MAT procedure.


An experimental analysis of sociocultural variables in sales of cigarettes to minors.
This study assessed the role of age, racial/ethnic group, and gender, as well as that of other sociocultural variables, in minors' access to tobacco. Thirty-six minors attempted to purchase cigarettes once in each of 72 stores (2592 purchase attempts). The minors represented equal numbers of girls and boys; 10-year-olds, 14-year-olds, and 16-year-olds; and Whites, Blacks, and Latinos. Equal numbers of stores were in Black, White, and Latino neighborhoods. Older children were more likely than younger ones to be sold cigarettes, and Latino children were more likely than Whites to be sold cigarettes. Older Black children (irrespective of gender) were the single most likely group to be sold cigarettes. Cigarettes were significantly more likely to be sold to children by male than female clerks and in specific sociocultural contexts. Interventions with retailers must address sociocultural variables to improve effectiveness in reducing minors' access to tobacco.


[Use of a preterm formula with long chain polyunsaturated fatty acids: acceptability and tolerance].
The majority of preterm formulas have included long chain polyunsaturated fatty acids (LCPUFAS) for the relationship with visual and cognoscitive brain development. This work tried to probe the tolerance and acceptability of a preterm formula from the point of view of palatability for the LCPUFAs origin. Infants under 37 weeks of gestational age at birth and birth weight below 2,500g with the possibility of being fed by suction and not receiving breast milk were included. Using a scale modified from "Pain Analog Scale," the nurse who was feeding the infant had to qualify the baby's facial expression at these moments. 50 infants with age 37.2 +/- days of life at least receiving the formula for 3 complete days were included. Ninety percent of them had weight gain each day and the third day was the most. There were no problems in number and frequency of stools. Thus, 782 evaluations by the nurses were made (15 for each patient); 66.8% of the time the infants accepted the formula with no problem; 21.1% were satisfactory; 3.7% were very satisfactory. Also, 91.8% of the nurses declared their satisfaction with the formula, and in 4 cases they declared dissatisfaction. They attributed it to the patient's pathology.


[Evidence-based nuclear medicine and experience-based nuclear medicine].
The basis and methods of Evidence-Based medicine (EBM) and its applications in Nuclear Medicine (NM) are described. EBM criteria are contrasted with clinical experience. Some NM Meta-analysis and Systematic Reviews are described. The methods of redefining the probability through diagnostic tests and the methods of Decision Analysis and of Cost-Efficiency Studies applied to NM are described. Finally a Series of proposals to improve the application of EBM to NM are presented.


Hypersensitive sites in the 5' and 3' flanking regions of the cysteine proteinase I gene of Dictyostelium discoideum.
The cysteine proteinase I gene of Dictyostelium discoideum is a developmentally regulated single copy gene. Specific sites in the 5' and the 3' flanking regions of the gene were cleaved by an endogenous nuclease when the gene was being transcribed. The majority of these sites were not cut when the gene was inactive. A dramatic change in the pattern of micrococcal nuclease and DNase I hypersensitive sites occurred in the 5' flanking region when transcription commenced at the 8 h stage of development. The major sites, doublets at -220/-300 bp and -670/-770 bp upstream of the transcription start site, corresponded to those cut by the endogenous nuclease. When transcription subsequently ceased the hypersensitive sites did not significantly change, indicating the gene remained in an activated state. The micrococcal nuclease hypersensitive sites in the 3' flanking region did not change significantly during development.


Seeking treatment for uncomplicated malaria: experiences from the Kintampo districts of Ghana.
Malaria accounts for many deaths and illnesses, mostly among young children and pregnant women in sub-Saharan Africa. An integrated approach is recommended to ensure effective malaria control. Socio-cultural factors continue to serve as determinants of malaria health-seeking behaviour. An INDEPTH effectiveness and safety study platform was established to unearth issues around the use of licensed and nationally recommended anti-malarials in real life settings. This study reports on treatment-seeking behaviour for uncomplicated malaria among community members. A qualitative study was conducted in the dry and rainy seasons in purposively selected communities in Kintampo north and south districts. This was based on distances to a health facility, ethnicity and availability of medicines at the sale outlets. Twenty-four focus group discussions were conducted among adult men, women care-takers of children less than 5 years and pregnant women. Ten INDEPTH interviews were also conducted among operators of medicine sale outlets and managers of health facilities. Fifty-one illnesses narrative interviews were conducted among adult men, women, women caretakers of children less than 5 years and pregnant women. Transcripts were transferred into Nvivo 8 software for data management and analysis. The artemisinin-based combinations that were commonly known and used were artesunate-amodiaquine and artemether-lumefantrine. Use of herbal preparation to treat diseases including uncomplicated malaria is rife in the communities. Drug stores were not the main source of artemisinin-based combination sales at time of the study. Monotherapies, pain killers and other medicines were purchased from these shops for malaria treatment. Dizziness, general body weakness and sleepiness were noted among respondents who used artemisinin-based combination therapy (ACT) in the past. There is no clear cut trajectory for management of uncomplicated malaria in the study area. Different approaches are adopted when treating malaria. There is need for community education to influence behaviour on the management of malaria to achieve real gains from ACT use.


Dynamic critical behavior of the XY model in small-world networks.
The critical behavior of the XY model on small-world network is investigated by means of dynamic Monte Carlo simulations. We use the short-time relaxation scheme, i.e., the critical behavior is studied from the nonequilibrium relaxation to equilibrium. Static and dynamic critical exponents are extracted through the use of the dynamic finite-size scaling analysis. It is concluded that the dynamic universality class at the transition is of the mean-field nature. We also confirm numerically that the value of dynamic critical exponent is independent of the rewiring probability P for P greater, similar 0.03.


Comparison of two intravitreal ranibizumab treatment schedules for neovascular age-related macular degeneration.
Ranibizumab (Lucentis), a humanised antibody fragment that inhibits vascular endothelial growth factor (VEGF)-A, is widely used for the treatment of neovascular age-related macular degeneration (NV-AMD). The objective of this study was to compare the outcomes of two different treatment protocols: loading dose (LD) and pro re nata (PRN (as needed)) dosing schedule from baseline. This retrospective chart review was conducted at King's College Hospital, London, UK. Consecutive patients were identified using the 'Ranibizumab in NV-AMD' database. These patients had treatment-naive choroidal neovascularisation (CNV) secondary to AMD, received ranibizumab therapy and had completed 12 months of follow-up. Baseline examination included visual acuity (Early Treatment Diabetic Retinopathy Study (ETDRS) letters), slit-lamp biomicroscopy, fluorescein angiography, and qualitative and quantitative assessment of central macular characteristics on optical coherent tomography (OCT). Intravitreal ranibizumab (0.5mg/0.05ml) was given to all patients at baseline. Patients on LD regimen received two further consecutive monthly intravitreal ranibizumab injections independent of clinical findings. Further injections were determined by the same re-treatment criteria as patients on PRN schedule from baseline. The main outcome variables in the two treatment groups were visual acuity and central macular thickness at different time points. The LD group contained 47 patients and the PRN group contained 31 patients. There were no significant differences between groups in the mean changes in visual acuity or central macular thickness. Visual acuity was similar in both groups at 6 months. However, twice as many patients improved visual acuity by 15 or more letters in the LD group (29.8% in the LD group compared with 12.9% in the PRN group (p=0.01)). This study showed that standard protocols used for OCT-guided retreatment achieved smaller mean gains in vision than those obtained with monthly ranibizumab administration. Further, loading doses of ranibizumab resulted in more visual gains than the PRN protocol.


Mutagenic fate of insecticide fenitrothion in the environment-mutagenicity increases both by anaerobic biodegradation and photodegradation.
In the present study, our objectives were (1) using the Ames assay, to evaluate the change in mutagenicity of a fenitrothion-containing solution during aerobic biodegradation, anaerobic biodegradation, and photodegradation, and (2) to identify possible mutagenic transformed products (TPs) that contributed substantially to any increase in mutagenicity. Mutagenicity of the fenitrothion-containing solution did not increase during aerobic biodegradation with any of the tested bacterial strains. In contrast, the mutagenicity increased for strain YG1029 during anaerobic biodegradation because of the generation of a strongly mutagenic TP, amino-fenitrothion. During photodegradation, mutagenicities increased slightly for YG1021 and YG1024, possibly owing to the production of a previously unreported mutagenic TP.


Reproduction Symposium: developmental programming of reproductive and metabolic health.
Inappropriate programming of the reproductive system by developmental exposure to excess steroid hormones is of concern. Sheep are well suited for investigating developmental origin of reproductive and metabolic disorders. The developmental time line of female sheep (approximately 5 mo gestation and approximately 7 mo to puberty) is ideal for conducting sequential studies of the progression of metabolic and/or reproductive disruption from the developmental insult to manifestation of adult consequences. Major benefits of using sheep include knowledge of established critical periods to target adult defects, a rich understanding of reproductive neuroendocrine regulation, availability of noninvasive approaches to monitor follicular dynamics, established surgical approaches to obtain hypophyseal portal blood for measurement of hypothalamic hormones, and the ability to perform studies in natural setting thereby keeping behavioral interactions intact. Of importance is the ability to chronically instrument fetus and mother for determining early endocrine perturbations. Prenatal exposure of the female to excess testosterone (T) leads to an array of adult reproductive disorders that include LH excess, functional hyperandrogenism, neuroendocrine defects, multifollicular ovarian morphology, and corpus luteum dysfunction culminating in early reproductive failure. At the neuroendocrine level, all 3 feedback systems are compromised. At the pituitary level, gonadotrope (LH secretion) sensitivity to GnRH is increased. Multifollicular ovarian morphology stems from persistence of follicles as well as enhanced follicular recruitment. These defects culminate in progressive loss of cyclicity and reduced fecundity. Prenatal T excess also leads to fetal growth retardation, an early marker of adult reproductive and metabolic diseases, insulin resistance, hypertension, and behavioral deficits. Collectively, the reproductive and metabolic deficits of prenatal T-treated sheep provide proof of concept for the developmental origin of fertility and metabolic disorders. Studies with the environmental endocrine disruptor bisphenol A (BPA) show that reproductive disruptions found in prenatal BPA-treated sheep are similar to those seen in prenatal T-treated sheep. The ubiquitous exposure to endocrine disrupting compounds with steroidogenic potential via the environment and food sources calls for studies addressing the impact of developmental exposure to environmental steroid mimics on reproductive function.


In vivo and in vitro biotransformation of the lithium salt of gamma-linolenic acid by three human carcinomas.
Lipid metabolism has been considered recently as a novel target for cancer therapy. In this field, lithium gamma-linolenate (LiGLA) is a promising experimental compound for use in the treatment of human tumours. In vivo and in vitro studies allowed us to assess the metabolism of radiolabelled LiGLA by tumour tissue and different organs of the host. In vitro studies demonstrated that human pancreatic (AsPC-1), prostatic (PC-3) and mammary carcinoma (ZR-75-1) cells were capable of elongating GLA from LiGLA to dihomo-gamma-linolenic acid (DGLA) and further desaturating it to arachidonic acid (AA). AsPC-1 cells showed the lowest delta5-desaturase activity on DGLA. In the in vivo studies, nude mice bearing the human carcinomas were given Li[1-(14)C]GLA (2.5 mg kg(-1)) by intravenous injection for 30 min. Mice were either sacrificed after infusion or left for up to 96 h recovery before sacrifice. In general, the organs showed a maximum uptake of radioactivity 30 min after the infusion started (t = 0). Thereafter, in major organs the percentage of injected radioactivity per g of tissue declined below 1% 96 h after infusion. In kidney, brain, testes/ovaries and all three tumour tissues, labelling remained constant throughout the experiment. The ratio of radioactivity in liver to tumour tissues ranged between 16- and 24-fold at t = 0 and between 3.1- and 3.7-fold at 96 h. All tissues showed a progressive increase in the proportion of radioactivity associated with AA with a concomitant decrease in radiolabelled GLA as the time after infusion increased. DGLA declined rapidly in liver and plasma, but at a much slower rate in brain and malignant tissue. Seventy-two hours after the infusion, GLA was only detected in plasma and tumour tissue. The sum of GLA + DGLA varied among tumour tissues, but it remained 2-4 times higher than in liver and plasma. In brain, DGLA is the major contributor to the sum of these fatty acids. Data showed that cytotoxic GLA and DGLA, the latter provided either by the host or by endogenous synthesis, remained in human tumours for at least 4 days.


Effect of trace elements on citric acid fermentation by Aspergillus niger.
Citric acid yields of 98.7% (sugar consumption basis) were reached in shaker flasks with mutant UV-ET-71-15 of Aspergillus niger in a resin-treated sucrose medium of the following composition (g/100 ml): sucrose, 14.0; NH(4)NO(3), 0.20; KH(2)PO(4), 0.10; MgSO(4).7H(2)O, 0.025; and (mg/liter): FeSO(4), 0.15 to 0.75; ZnSO(4), 0.10; and CuSO(4), 0.01. Yields of 75% were obtained in medium with resin-treated clarified syrup and 68% with ferrocyanide-treated blackstrap molasses. Optimal conditions included selection of appropriate pellets as inoculum at 3%, pH of 4.5, temperature at 30 C, agitation at 250 rev/min, and fermentation time of 8 days. The mutant tolerated high concentrations of trace elements.


Inflammatory biomarkers and oxidative stress measurements in patients with systemic lupus erythematosus with or without metabolic syndrome.
The aims of the present study were to report the frequency of metabolic syndrome in systemic lupus erythematosus (SLE); to verify differences in inflammatory biomarkers and oxidative stress in SLE patients with or without metabolic syndrome; and to assess which metabolic syndrome components are associated with oxidative stress and disease activity. The study included 58 SLE patients and 105 controls. SLE patients were divided in two groups, with and without metabolic syndrome. 41.4% patients met the criteria for metabolic syndrome compared with 10.5% controls. Patients with SLE and metabolic syndrome had significantly raised serum uric acid, C-reactive protein (CRP), lipid hydroperoxides, and protein oxidation when compared with patients with SLE without metabolic syndrome. Lipid hydroperoxides were correlated with CRP, whereas protein oxidation was associated with waist circumference and uric acid. There was a positive association between serum C3 and C4 and glucose and between C3 and CRP. SLE disease activity index (SLEDAI) scores were positively correlated with body mass index (BMI) and waist circumference (WC). In conclusion, SLE patients have a high prevalence of metabolic syndrome and this syndrome directly contributes to increase inflammatory status and oxidative stress. Inflammatory processes, being overweight/obese, and uric acid may favor oxidative stress increases in patients with SLE and metabolic syndrome. C3 and C4 may have a positive acute-phase protein behavior in patients with SLE.


The perturbation of thrombin binding to human platelets by anions.
Thrombin binds with high affinity to specific cell-surface receptors on washed human platelets. We present experiments indicating that thrombin binding correlates withe the release reaction when binding is perturbed by anions. Marked differences in the affinity of human 125I-thrombin for platelets wer observed in various isotonic buffers at pH 7.4. At low concentrations of thrombin (0.001-0.01 U/ml), binding was 5-fold greater in Tris-sodium acetate and 12-fold greater in Tris-sodium cacodylate than in Tris-sodium chloride. These anion-induced changes in 125I-thrombin binding paralleled changes in [14C] serotonin release when both parameters were measured in the same platelets. Thus, equivalent release occurred for equal amounts of thrombin bound in all buffers, even though the thrombin concentration varied by up to 30-fold. After approximately 100 molecules of thrombin bound per platelet, complete release occurred in all buffers in 2 min. The effect of anions was specific for the thrombin-receptor interaction as there was no corresponding effect on the binding of erythroagglutinating phytohemagglutinin (E-PHA) to platelets nor on E-PHA or collagen-induced serotonin release. The various anions did not alter platelet morphology as judged by electron microscopy. The anions had no effect on thrombin esterase catalytic activity. In addition, the total number of thrombin receptors per platelet was approximately the same in all buffers. Thus anions alter the affinity between platelet thrombin receptors and a site on thrombin distinct from the catalytic site. We conclude that the thrombin receptor is essential for thrombin-induced platelet reactions.


The clubfoot over the centuries.
The aim of this paper is to review all treatment methods of the clubfoot over the years through the documentation present in the literature and art with the aim of better understanding the pathoanatomy of the deformity, but to also clarify factors that allow a safe, logical approach to clubfoot management. The initial part of this paper traces the most representative iconographic representations of clubfoot in history to describe how his presence was witnessed since ancient times. Hippocrates, the father of medicine, was the first to present written references about treatment methods of clubfoot. After the dark period of the Middle Ages, during the Renaissance, there were more detailed studies on the disease, with the creation of the first orthopedic orthotics, designed to correct the deformity with the help of famous names in medicine's history. In the XVIII century, as we witness the birth of orthopedics as a distinct discipline in medicine, more and more brilliant minds developed complex orthoses and footwear with the aim of obtaining a proper correction of the deformity. In the last part of the paper, there is a description of the main surgical techniques developed over the years until the return to conservative treatment methods such as the Ponseti method, internationally recognized as the gold standard of treatment, despite the presence of some unresolved issues such as the possible recurrence of the disease.


Emerging directions in the study of the ecology and evolution of plant-animal mutualistic networks: a review.
The study of mutualistic plant and animal networks is an emerging field of ecological research. We reviewed progress in this field over the past 30 years. While earlier studies mostly focused on network structure, stability, and biodiversity maintenance, recent studies have investigated the conservation implications of mutualistic networks, specifically the influence of invasive species and how networks respond to habitat loss. Current research has also focused on evolutionary questions including phylogenetic signal in networks, impact of networks on the coevolution of interacting partners, and network influences on the evolution of interacting species. We outline some directions for future research, particularly the evolution of specialization in mutualistic networks, and provide concrete recommendations for environmental managers.


Adaptation of dynamic covalent systems of imine constituents to medium change by component redistribution under reversible phase separation.
A dynamic covalent library of interconverting imine constituents, dissolved in an acetonitrile/water mixture, undergoes constitutional reorganization upon phase separation induced by a physical stimulus (heat) or a chemical effector (inorganic salt, carbohydrate, organic solvent). The process has been made reversible, regenerating the initial library upon phase reunification. It represents the behavior of a dynamic covalent library upon reversible phase separation and its adaptation to a phase change, with up-regulation in each phase of the fittest constituents by component selection. Finally, the system exemplifies the splitting of a 2D (square) constitutional dynamic network into a 3D (cube) one.


Paratesticular Dedifferentiated Liposarcoma with Rhabdomyoblastic Differentiation: A Case Report and Review of the Literature.
Liposarcomas of the paratesticular tissue is a rare pathological entity. The symptoms are similar to inguinal hernias or hydroceles. We present the case of an 84-year-old man with a rare paratesticular liposarcoma that manifested as painless right hemiscrotal swelling. Testicular tumour markers were negative. Imaging revealed a heterogeneous mass with a fat component. He underwent a radical orchiectomy on the left side to remove the associated mass. This revealed dedifferentiated liposarcoma (DDLS) with rhabdomyoblastic differentiation and MDM2 amplification. The surgical margins were negative, and the patient had a metastatic workup that included magnetic resonance imaging (MRI) of the abdomen and pelvis. Because of the disease's rarity, there is no clear agreement on radiotherapy and chemotherapy roles.