A mitochondrial Hsp70 orthologue in Vairimorpha necatrix: molecular evidence that microsporidia once contained mitochondria.
Microsporidia are small (1-20 micron) obligate intracellular parasites of a variety of eukaryotes, and they are serious opportunistic pathogens of immunocompromised patients [1]. Microsporidia are often assigned to the first branch in gene trees of eukaryotes [2,3], and are reported to lack mitochondria [2,4]. Like diplomonads and trichomonads, microsporidia are hypothesised to have diverged from the main eukaryotic stock prior to the event that led to the mitochondrion endosymbiosis [2,4]. They have thus assumed importance as putative relics of premitochondrion eukaryote evolution. Recent data have now revealed that diplomonads and trichomonads contain genes that probably originated from the mitochondrion endosymbiont [5-9], leaving microsporidia as chief candidates for an extant primitively amitochondriate eukaryote group. We have now identified a gene in the microsporidium Vairimorpha necatrix that appears to be orthologous to the eukaryotic (symbiont-derived) Hsp70 gene, the protein product of which normally functions in mitochondria. The simplest interpretation of our data is that microporidia have lost mitochondria while retaining genetic evidence of their past presence. This strongly suggests that microsporidia are not primitively amitochondriate and makes feasible an evolutionary scenario whereby all extant eukaryotes share a common ancestor which contained mitochondria.


The Combination of Computational and Biosensing Technologies for Selecting Aptamer against Prostate Specific Antigen.
Herein, we report a method of combining bioinformatics and biosensing technologies to select aptamers against prostate specific antigen (PSA). The main objective of this study is to select DNA aptamers with higher binding affinity for PSA by using the proposed method. Based on the five known sequences of PSA-binding aptamers, we adopted the functions of reproduction and crossover in the genetic algorithm to produce next-generation sequences for the computational and experimental analysis. RNAfold web server was utilized to analyze the secondary structures, and the 3-dimensional molecular models of aptamer sequences were generated by using RNAComposer web server. ZRANK scoring function was used to rerank the docking predictions from ZDOCK. The biosensors, the quartz crystal microbalance (QCM) and a surface plasmon resonance (SPR) instrument, were used to verify the binding ability of selected aptamer for PSA. By carrying out the simulations and experiments after two generations, we obtain one aptamer that can have the highest binding affinity with PSA, which generates almost 2-fold and 3-fold greater measured signals than the responses produced by the best known DNA sequence in the QCM and SPR experiments, respectively.


Integration, capitation, and managed care: transformation of nursing for 21st century health care.
Market-driven reforms and the growth of managed care will transform the nursing profession in the next 5 to 10 years. Managed care will pressure RN-level nursing to justify its costs and advanced training. New roles for nursing will include (a) advanced practice nurses substituting for physicians, (b) nurse case managers coordinating care and aggressively managing patients along care paths, and (c) nurse team leaders organizing patient care and delegating less complex nurse tasks to less trained workers.


Red blood cell as glucose carrier: significance for placental and cerebral glucose transfer.
At plasma glucose values of 5 mM (90 mg/100 ml) the maximum glucose transport capacity of the human red cell membrane is 12,000 times the rate of glucose utilization by the red blood cell. Mammals, other than primates, that have been tested have a comparable high-capacity system during fetal life, which is lost soon after birth. It has been suggested that the availability of the water space of the red blood cell for distribution of glucose facilitates transfer across the placenta during fetal life in all mammals and across the blood-brain barrier in adult primates. Though plausible, more comparative studies of glucose transport in red blood cells of other species and direct experimental evaluations of the contribution of the red blood cell to glucose transfer across the placenta and the blood-brain barrier are needed.


[Epidemiological enquiries in two Q fever outbreaks in a community of Baden-Württemberg during 2008 and 2009].
In 2008 and 2009, two consecutive outbreaks of Q fever in humans were recorded in the district of Freudenstadt, northern Black Forrest, Baden-Württemberg, Germany. In 2008, a total of 41 persons from a single local community fell ill and were found infected with Coxiella burnetii. Although comprehensive diagnostic and epidemiological outbreak investigations were conducted and control measures taken which included vaccination of ruminants at risk in three parts of the affected community, re-occurrence of the disease in 2009 with further 29 confirmed human Q fever cases could not be prevented. While the origin of infection of the first outbreak was probably a flock of 550 sheep moved in the surrounding of the affected villages, the source of infection for the consecutive outbreak in 2009 could not be identified. It seems possible that meadows contaminated with infectious placenta or birth fluids represented the sources of infection.


Effects of mobile phone electromagnetic fields on brain waves in healthy volunteers.
The interaction between biological tissue and electromagnetic fields (EMF) is a topic of increasing interest due to the rising prevalence of background EMF in the past decades. Previous studies have attempted to measure the effects of EMF on brainwaves using EEG recordings, but are typically hampered by experimental and environmental factors. In this study, we present a framework for measuring the impact of EMF on EEG while controlling for these factors. A Bayesian statistical approach is employed to provide robust statistical evidence of the observed EMF effects. This study included 32 healthy participants in a double-blinded crossover counterbalanced design. EEG recordings were taken from 63 electrodes across 6 brain regions. Participants underwent a measurement protocol comprising two 18-min sessions with alternating blocks of eyes open (EO) and eyes closed (EC) conditions. Group 1 (n = 16) had EMF during the first session and sham during the second session; group 2 (n = 16) had the opposite. Power spectral density plots were generated for all sessions and brain regions. The Bayesian analysis provided statistical evidence for the presence of an EMF effect in the alpha band power density in the EO condition. This measurement protocol holds potential for future research on the impact of novel transmission protocols.


Electrical, thermal and noise properties of platinum-carbon free-standing nanowires designed as nanoscale resistive thermal devices.
Platinum-carbon (PtC) composite nanowires were fabricated using focused electron beam induced deposition (FEBID) and postprocessed, and their performance as a nanoscale resistive thermal device (RTD) was evaluated. Nanowires were free-standing and deposited on a dedicated substrate to eliminate the influence of the substrate itself and of the halo effect on the results. The PtC free-standing nanowires were postprocessed to lower their electrical resistance using electron beam irradiation and thermal annealing using Joule heat both separately and combined. Postprocessed PtC free-standing nanowires were characterized to evaluate their noise figure (NF) and thermal coefficients at the temperature range from 30°C to 80°C. The thermal sensitivity of RTD was lowered with the reduced resistance but simultaneously the NF improved, especially with electron-beam irradiation. The temperature measurement resolution achievable with the PtC free-standing nanowires was 0.1 K in 1 kHz bandwidth.


Food security in HIV/AIDS response: insights from Homa Bay, Kenya.
This paper examines the viability and effectiveness of a pilot farming initiative in reversing impacts of HIV/AIDS on the most affected households in Homa Bay, Kenya. The paper argues that once patients are stable, they can effectively be engaged in farming with minimal financial and technical support, resulting in enhanced food security of the affected households. More importantly, it helps to reduce HIV/AIDS-related stigma and improve the individual's self-esteem. Some of the key challenges of the pilot initiative were the limited number of agricultural extension workers and absence of facilities to enable them to deliver services to the farmers, the high cost of farm inputs, the unavailability of farm inputs when they were needed, poorly developed agricultural markets, and the absence of irrigation facilities. The paper recommends the sensitive scaling-up of this approach. However, farming initiatives by HIV/AIDS service NGOs should be linked to at least three key aspects: (a) treatment, care and support to HIV/AIDS affected households; (b) micro grant schemes or subsidies to enable farmers to purchase farming tools and farm inputs; and (c) comprehensive on-farm training support. To ensure effectiveness and wider reach, government needs to view agriculture through an HIV lens and promote a multisectoral approach that recognises the relationship between HIV/AIDS and food security. A number of immediate actions are required to strengthen this relationship, such as increased public investment to augment extension services, subsidise farm inputs, and develop infrastructure including agricultural markets.


Glucocorticoid administration in athletes: Performance, metabolism and detection.
It is generally acknowledged in the sporting world that glucocorticoid (GC) use enhances physical performance. This pharmacological class is therefore banned by the World Anti-Doping Agency (WADA) in in-competition samples after systemic but not local (defined as any route other than oral, intravenous, intramuscular or rectal) administration, which thus allows athletes to use GCs for therapeutic purposes. According to the 2016 WADA list, the urine reporting level for all GCs is set at 30ng/ml to distinguish between the authorized and banned routes of administration. The actual data on the ergogenic effects of GC intake are nevertheless fairly recent, with the first study showing improved physical performance with systemic GC administration dating back only to 2007. Moreover, the studies over the last decade coupling ergogenic and metabolic investigations in humans during and after GC intake have shown discrepant results. Similarly, urine discrimination between banned and authorized GC use remains complex, but it seems likely to be improved thanks to new analytical studies and the inclusion of the authorized GC uses (local routes of administration and out-of-competition samples) in the WADA monitoring program. In this review, we first summarize the current knowledge on the ergogenic and metabolic GC effects in humans during various types of exercise. We then present the antidoping legislation and methods of analysis currently used to detect GC abuse and conclude with some practical considerations and perspectives.


Infant mortality among singletons and twins in Japan during 1999-2008 on the basis of risk factors.
The infant mortality rate (IMR) among single and twin births from 1999 to 2008 was analyzed using Japanese Vital Statistics. The IMR was 5.3-fold higher in twins than in singletons in 1999 and decreased to 3.9-fold in 2008. The reduced risk of infant mortality in twins relative to singletons may be related, partially, to survival rates, which improved after fetoscopic laser photocoagulation for twin - twin transfusion syndrome. The proportion of neonatal deaths among total infant deaths was 54% for singletons and 74% for twins. Thus, intensive care of single and twin births may be very important during the first month of life to reduce the IMR. The IMR decreased as gestational age (GA) rose in singletons, whereas the IMR in twins decreased as GA rose until 37 weeks and increased thereafter. The IMR was significantly higher in twins than in singletons from the shortest GA (<24 weeks) to 28 weeks as well as ≥38 weeks, whereas the IMR was significantly higher in singletons than in twins from 30 to 36 weeks. As for maternal age, the early neonatal and neonatal mortality rates as well as the IMR in singletons were significantly higher in the youngest maternal age group than in the oldest one, whereas the opposite result was obtained in twins. The lowest IMR in singletons was 1.1 per 1,000 live births for ≥38 weeks of gestation and heaviest birth weight (≥2,000 g), while the lowest IMR in twins was 1.8 at 37 weeks and ≥2,000 g.


Pattern Analysis of Inpatient Falls in a Tertiary Hospital.
We analyzed the spatial and temporal patterns of inpatient falls in a tertiary hospital. Data were obtained from the adverse-event self-reporting system and the electronic nursing record system of the hospital. Through chart reviews we reclassified the cases. Most falls occurred by the bedside, followed by the restroom and hallway, and they occurred most often between 2 a.m. and 6 a.m. These findings indicate when and where nurses should be most alert about falls.


Thrombospondin-1 opens the paracellular pathway in pulmonary microvascular endothelia through EGFR/ErbB2 activation.
Thrombospondin-1 (TSP1) is a multidomain protein that contains epidermal growth factor (EGF)-like repeats that indirectly activate the EGF receptor (EGFR) and selected downstream signaling pathways. In these studies, we show that TSP1 opens the paracellular pathway in human lung microvascular endothelial cells (HMVEC-Ls) in a dose-, time-, and protein tyrosine kinase (PTK)-dependent manner. TSP1 increased tyrosine phosphorylation of proteins enriched to intercellular boundaries including the zonula adherens (ZA) proteins, vascular endothelial-cadherin, γ-catenin, and p120 catenin. In HMVEC-Ls, EGFR and ErbB2 are expressed at low levels, and both heterodimerize and tyrosine autophosphorylate in response to TSP1. Prior EGFR-selective PTK inhibition with AG1478 or ErbB2-selective PTK inhibition with AG825 protected against TSP1-induced tyrosine phosphorylation of ZA proteins and barrier disruption. Preincubation of HMVEC-Ls with an EGFR ectodomain-blocking antibody also prevented TSP1-induced opening of the paracellular pathway. Therefore, in HMVEC-Ls, TSP1 increases tyrosine phosphorylation of ZA proteins and opens the paracellular pathway, in part, through EGFR/ErbB2 activation. Surprisingly, recombinant TSP1 EGF-like repeats 1-3 and the high-affinity EGFR ligands, EGF, TGF-α, and amphiregulin, each failed to increase paracellular permeability. However, HMVEC-Ls in which EGFR was overexpressed became responsive to the EGF-like repeats of TSP1 as well as to EGF. These studies indicate that TSP1 disrupts the endothelial barrier through EGFR/ErbB2 activation although additional signals are necessary in cells with low receptor expression.


Toxicity of cocaine.
Abuse of central nervous system (CNS) stimulants continues to escalate within the United States. This group of powerful drugs includes cocaine, "crack" (free-base cocaine), methamphetamine, and "ice" (free-base methamphetamine). The pharmacological actions are similar among these agents, but their onsets and durations of action differ. Patients using and abusing cocaine and methamphetamines may present themselves as dental patients. It is therefore important for dental clinicians to understand the pharmacology of these drugs.


An examination of barriers to nurse practitioner endorsement in senior rural drug and alcohol nurses in New South Wales.
To examine barriers senior rural nurses in New South Wales drug and alcohol clinical settings perceive when considering endorsement to Nurse Practitioner (NP). A survey was designed to record views of senior alcohol and drug nurses in rural New South Wales about becoming a NP. Participants were identified by Area Health Service Directors of Drug and Alcohol Services for each Area Health Service in NSW excluding metropolitan Sydney. Forty eight surveys were distributed, with 17 (35%) completed surveys included in the results. Of the 17 participants, 12 (70.6%) expressed interest in becoming a NP. The majority (12, 70.6%) were unaware or unsure of endorsement processes. Lack of clarity about the NP role and processes and benefits to becoming a NP was found to be of most concern to all participants. Only 6 participants (35%) indicated they would consider seeking endorsement to NP. Despite agreeing that NP positions in alcohol and drug settings would improve patient access to treatment and care, senior nurses working in these settings in regional areas are unfamiliar with pathways to becoming endorsed. Barriers, such as a lack of internal support from management and colleagues, as well as the fact that respondents reported no foreseeable financial gain in endorsement, also need to be addressed, before more nurses will consider endorsement. Further evaluation of the views of senior nurses in metropolitan alcohol and other drug settings in the process to NP endorsement is needed before clients will benefit from the expertise and enhanced care that NP's may provide.


Fibrogenesis and fibrosis in inflammatory bowel diseases: Good and bad side of same coin?
Fibrogenesis in inflammatory bowel diseases is a complex phenomenon aimed at mucosal repair. However, it may provoke intestinal fibrosis with the development of strictures which require surgery. Therefore, fibrogenesis may be considered as a "two-faced" process when related to chronic intestinal inflammation. Many types of cells may be converted into the fibrogenic phenotype at different levels of the intestinal wall. A complex interaction of cytokines, adhesion molecules and growth factors is involved in the process. We report an overview of recent advances in molecular mechanisms of stricturizing Crohn's disease (CD) including the potential role of trasforming growth factor beta, protein kinase C and Ras, Raf and ERK proteins. Fibrotic growth factors such as vascular endothelial growth factor and platelet-derived growth factor, as well as the Endothelial-to-Mesenchymal Transition induced by transforming growth factor-β, are considered. Finally, our experience, focused on tumor necrosis factor α (the main cytokine of inflammatory bowel diseases) and the link between syndecan 1 (a heparan sulphate adhesion molecule) and basic fibroblast growth factor (a strong stimulator of collagen synthesis) is described. We hypothesize a possible molecular pattern for mucosal healing as well as how its deregulation could be involved in fibrotic complications of CD. A final clinical point is the importance of performing an accurate evaluation of the presence of fibrotic strictures before starting anti-tumor necrosis α treatment, which could worsen the lesions.


Inter-university cooperation for establishing an M.Sc. course in health informatics aims at contributing to users' acceptance of health telematics applications.
Health informatics is a recently established and important multi-disciplinary and inter-disciplinary field that not only involves informatics but also medicine, nursing, engineering, biology and other-related subjects. A coordination of this field at a postgraduate level becomes important now in Europe where other European Community programs such as the Telematics for Health Care will require at the Fourth Framework Programme (1994-1999) and the Fifth Framework Programme (2000-2006) adequate human resources of higher potential and knowledge. A European M.Sc. course met all the above objectives. The curriculum was developed according to previous experiences in similar programmes. Recently the course has been organised on the basis of an Inter-University nature with the participation of 5 Greek Universities. The paper aims at providing a description of the new academic programme and a brief evaluation of the implementation phase.


The effect of growth conditions on adenylate cyclase activity and virulence-related properties of Bordetella pertussis.
Growth of Bordetella pertussis in Stainer & Scholte medium in which the NaCl had been replaced by one of several inorganic or organic salts resulted in a large decrease in adenylate cyclase activity, histamine-sensitizing activity and in the amounts of two cell-envelope polypeptides of Mr 28000 and 30000. Although some variation between strains was observed, there was never a case where one of these properties was lost independently of the others. Cultures in which these properties were lost had decreased amounts of extracellular cAMP when compared to NaCl-grown cultures. Adenylate cyclase activity was detected in three locations of B. pertussis cultures (extracellular, extracytoplasmic but cell-associated, and cytoplasmic). After growth in medium containing high concentrations of MgSO4, enzyme activity was decreased to a similar extent in all three locations.


Widening the sphere of influence: using a tool to extend extrapersonal visual space in a patient with severe neglect.
We report evidence that visual representations of space close to the body can be extended when a patient uses a tool to explore the environment. HB had severe neglect of left and far spatial regions which was determined more by how locations were visually perceived than by how they were represented tactilely or through proprioception. His ability to detect visual targets in left and far space was improved, however, when he held a tool. He also had limited tactile/proprioceptive knowledge about the location of his hand. These data suggest that by holding a tool, HB's more intact representation of near, visual space could be extended to include stimuli presented at a distance from his body. This extension of space improved his detection of visual stimuli. We discuss the implications of the results for the nature of our internal representation of space.


Effect of abutment modification and cement type on retention of cement-retained implant supported crowns.
Provisional cements are commonly used to facilitate retrievability of cement-retained fixed implant restorations; but compromised abutment preparation may affect the retention of implant-retained crowns.The purpose of this study was to investigate the effect of abutment design and type of luting agent on the retentive strength of cement-retained implant restorations. Two prefabricated abutments were attached to their corresponding analogs and embedded in an acrylic resin block. The first abutment (control group) was left intact without any modifications. The screw access channel for the first abutment was completely filled with composite resin. In the second abutment, (test group) the axial wall was partially removed to form an abutment with 3 walls. Wax models were made by CAD/CAM. Ten cast copings were fabricated for each abutment. The prepared copings were cemented on the abutments by Temp Bond luting agent under standardized conditions (n=20). The assemblies were stored in 100% humidity for one day at 37°C prior to testing. The cast crown was removed from the abutment using an Instron machine, and the peak removal force was recorded. Coping/abutment specimens were cleaned after testing, and the testing procedure was repeated for Dycal luting agent (n=20). Data were analyzed with two- way ANOVA (α=0.05). There was no significant difference in the mean transformed retention (Ln-R) between intact abutments (4.90±0.37) and the abutments with 3 walls (4.83±0.25) using Dycal luting agent. However, in TempBond group, the mean transformed retention (Ln-R) was significantly lower in the intact abutment (3.9±0.23) compared to the abutment with 3 walls (4.13±0.33, P=0.027). The retention of cement-retained implant restoration can be improved by the type of temporary cement used. The retention of cast crowns cemented to implant abutments with TempBond is influenced by the wall removal.


The Springer Model for Lifetime Prediction of Wind Turbine Blade Leading Edge Protection Systems: A Review and Sensitivity Study.
The wind energy sector is growing rapidly. Wind turbines are increasing in size, leading to higher tip velocities. The leading edges of the blades interact with rain droplets, causing erosion damage over time. In order to mitigate the erosion, coating materials are required to protect the blades. To predict the fatigue lifetime of coated substrates, the Springer model is often used. The current work summarizes the research performed using this model in the wind energy sector and studies the sensitivity of the model to its input parameters. It is shown that the Springer model highly depends on the Poisson ratio, the strength values of the coating and the empirically fitted a2 constant. The assumptions made in the Springer model are not physically representative, and we reasoned that more modern methods are required to accurately predict coating lifetimes. The proposed framework is split into three parts-(1) a contact pressure model, (2) a coating stress model and (3) a fatigue strength model-which overall is sufficient to capture the underlying physics during rain erosion of wind turbine blades. Possible improvements to each of the individual aspects of the framework are proposed.


A conformational epitope which detects autoantibodies from schizophrenic patients.
We previously found autoantibodies against platelets in schizophrenic patients. One of the platelet proteins that bind these antibodies is enolase. Here, we describe the isolation and sequencing of an immunoreactive peptide after enzymatic digestion of enolase. The 3-D structure of enolase indicates that, unexpectedly, this peptide is buried inside the protein. However, 3-D surface analysis leads to the identification of a conformational epitope that resembles the binding peptide and might constitute a specific binder of the autoantibodies. In a screening of antibody binding with the peptide LVVGLCK, we found in 50 serum samples of controls a mean of O.D.=0.46; s= +/- 0.21 relative enzyme immunoassay units, while in sera of 39 schizophrenic patients, we found a mean of O.D.=1.47; s= +/- 0.65; P<0.0001. Furthermore, an inverse correlation was observed between duration of schizophrenia and the level of the detected autoantibodies. A screening of autoantibodies in sera of various mental disorders with this peptide is currently in progress.


Innovative Characterization of Alternans Onset and Development in Dual Voltage-Calcium Whole-Heart Optical Mapping Signals at Multiple Thermal States.
Cardiac electrical dynamics show complex space-time instabilities, like period-doubling bifurcation and beat-to-beat alternans, known to occur as pro-arrhythmic phenomena and linked to membrane voltage and intracellular calcium kinetics. Besides, cellular ionic dynamics are critically affected by temperature oscillations, further enhancing the complexity of such arrhythmias precursors that lead to irregular cardiac contraction. In this complex scenario, fluorescence dual optical mapping techniques allow the unveiling of nonlinear and multi-scale couplings. In this contribution, we propose a novel methodological analysis of synchronous dual voltage-calcium traces obtained from whole rabbit hearts for (i) detecting alternans onset and evolution, (ii) characterizing novel restitution curves, and (iii) defining spatio-temporal alternans patterns at four thermal states. We validate our approach against well-accepted analyses considering complete pacing-down restitution protocols. The proposed methodology computes integral features, e.g., area under the curve, suggesting that a novel, easy-to-use indicator, may advance predictability on alternans onset and evolution, further providing insights into spatio-temporal cardiac analyses.Clinical Relevance- This work introduces new methods for the early detection of cardiac alternans onset and development as precursors of arrhythmias and fibrillation at different temperatures.


A comparison of rating and coding behavioural traits in dogs.
The aim of the present study was to examine the links between independent rating and coding approaches to assessing activity-impulsivity and inattention in dogs. Fifty-six adult Belgian shepherd dogs were videotaped performing in behavioural tests. Seventeen behavioural variables were measured by coders (video coding). Raters watched the same videotapes and then rated the activity-impulsivity and inattention of each dog (video rating). Owners filled out the Dog ADHS-RS questionnaire measuring activity-impulsivity and inattention. Video rating of activity-impulsivity correlated with the scale scores of the owner, but video codings did not. The results suggest that the owner ratings and video ratings are tapping the same constructs, but behavioural variables assessed in the present study were not appropriate for mirroring the owners' assessments. The findings suggest that if consistent individual differences in broad behavioural traits are the primary focus of analyses, then ratings seem to capture information not easily captured in coding approaches designed to assess the same constructs.


Drugs modulate allosterically heme-Fe-recognition by human serum albumin and heme-fe-mediated reactivity.
Human serum albumin (HSA) represents an important determinant of plasma oncotic pressure and a relevant factor that modulates fluid distribution between the body compartments. Moreover, HSA (i) represents the depot and transporter of several compounds, both endogenous and exogenous, (ii) affects the pharmacokinetics of many drugs, (iii) regulates chemical modifications of some ligands, (iv) shows (pseudo-)enzymatic properties, (v) inactivates some toxic compounds, and (vi) displays anti-oxidant properties. HSA binding and (pseudo-)enzymatic properties are regulated competitively, allosterically, and by covalent modifications. While competitive inhibition of HSA binding properties is evident, allosteric mechanisms and covalent modifications affecting HSA reactivity are less clear. In several pathological conditions in which free heme-Fe levels increase, the buffering capacity of plasma hemopexin is overwhelmed and most of heme-Fe binds to the fatty acid site 1 of HSA. HSA-heme-Fe displays globin-like properties; in turn, heme-Fe modulates competitively and allosterically HSA binding and reactivity properties. Remarkably, heme-Fe-mediated HSA properties are time-dependent, representing a case for "chronosteric effects". Here, we review the drug-based modulation of (i) heme-Fe-recognition by HSA and (ii) heme-Fe-mediated reactivity.


Metal biosorption by surface-layer proteins from Bacillus species.
Bacillus species have been involved in metal association as biosorbents, but there is not a clear understanding of this chelating property. In order to evaluate this metal chelating capacity, cultures and spores from Grampositive bacteria of species either able or unable to produce surface layer proteins (S-layers) were analyzed for their capacity of copper biosorption. Only those endowed of S-layers, like Bacillus sphaericus and B. thuringiensis, showed a significant biosorption capacity. This capacity (nearly 50%) was retained after heating of cultures, thus supporting that structural elements of the envelopes are responsible for such activity. Purified Slayers from two Bacillus sphaericus strains had the ability to biosorb copper. Copper biosorption parameters were determined for strain B. sphaericus 2362, and after analyses by means of the Langmuir model, the affinity and capacity were shown to be comparable to other bacterial biosorbents. A competitive effect of Ca2+ and Zn2+, but not of Cd2+, was also observed, thus indicating that other cations may be biosorbed by this protein. Spores that have been shown to be proficient for copper biosorption were further analyzed for the presence of Slayer content. The retention of S-layers by these spores was clearly observed, and after extensive treatment to eliminate the S-layers, the biosorption capacity of these spores was significantly reduced. For the first time, a direct correlation between S-layer protein content and metal biosorption capacity is shown. This capacity is linked to the retention of S-layer proteins attached to Bacillus spores and cells.


Human monoclonal antibodies directed against melanoma tumor-associated antigens.
To analyze the humoral immune response to melanoma, human-mouse hybridomas were generated by the fusions of regional lymph node lymphocytes of patients with the mouse myeloma cell line M5. Six stable hybridomas were cloned from six separate lymphocyte parents obtained from three patients. Ascites were obtained from nude mice after i.p. injection with cultured hybridoma cells. The monoclonal antibodies, four immunoglobulin Gs and two pentameric immunoglobulin Ms, were partially purified to remove mouse immunoglobulin and then conjugated to biotin for immunocytochemical and immunohistochemical studies. With the avidin:biotin:peroxidase complex method to detect and amplify binding by the biotin-conjugated human monoclonal antibodies, we found the six antibodies to be reactive against cytoplasmic determinants in five short-term melanoma cultures and formalin-fixed paraffin-embedded melanoma tumors from four patients. The antigenic target of the antibodies identified was not carcinoembryonic antigen. Two antibodies, 2-139-1 and 6-26-3, were studied in more detail. Each stained 25 of 25 specimens of melanomas. Little or no reactivity was detected against fixed sections of normal skin, which included tissues such as epidermis, dermis, monocytes, lymphocytes, and vascular endothelium. More striking was the absence of binding to melanocytes in the basal layer of the skin or to pigmented nevus cells. Both antibodies showed cross-reactivity against other tumors, in particular colonic and prostatic carcinomas. In the normal colon, reactivity was restricted to the surface of the columnar epithelium; no reactivity was detected against normal prostatic epithelium. Reactivity was also not observed against liver and lung. However, the epithelia of the renal tubules, pancreatic ducts, and salivary ducts were all reactive. These human monoclonal antibodies identify cytoplasmic melanoma-associated tumor antigens that appear different from the membrane antigens defined by serological approaches and by most mouse monoclonal antibodies.


Failure of quality control to detect errors in the preparation of technetium-99m disofenin (DISIDA).
Uptake of Tc-99m disofenin is so strongly hepatobiliary that striking appearance of radioactivity in the reticuloendothelial system (RES) is almost certainly caused by improper preparation of the disofenin kit. Four patients who received injections of Tc-99m disofenin demonstrated a pronounced RES biodistribution in addition to the expected hepatobiliary pattern. Since routine quality control testing had not forewarned us of this gross lack of radiochemical purity, Tc-99m disofenin kits were subjected to a variety of insults to elucidate the mechanisms of quality control failure. Assessment of radiochemical purity was conducted using instant thin-layer chromatography (ITLC) and two solvent systems. We concluded that both Tc-99m pertechnetate and Tc-99m sulfur colloid had been injected into the cold disofenin kit. Visual inspections were not reliable for verifying the final radiopharmaceutical product because of dilution of the sulfur colloid by pertechnetate. ITLC testing failed because the assay method being used at that time did not accurately assess the hydrolyzed fraction (colloidal component). Recommendations are made to preclude recurrence of the errors that occurred.


Influence of maternal dietary fat upon rat pups.
Mother rats were fed purified rations containing different fats during gestation and lactation and at 1 day after parturition. Litter sizes were reduced to 2 male and 2 female pups. The behavior and the brain chemical composition of these selected pups were compared with similarly selected pups from dams fed a commercial ration. All offspring were fed a commercial ration after weaning. Pups from dams fed 20% safflower oil were similar to controls. Feeding 20% cocoa butter to dams resulted in pups with reduced exploratory activity and with a rapid learning performance in a T-maze, employing the aversive stimulation of an electrical shock. Feeding dams a fat-free ration produced pups which had reduced rates of growth, small brains at 2 months of age, and low brain concentrations of cholesterol, DNA, and RNA.


Butyrate delivered by butyrylated starch increases distal colonic epithelial apoptosis in carcinogen-treated rats.
Animal studies show that increasing large bowel butyrate concentration through ingestion of butyrylated or resistant starches opposes carcinogen-induced tumorigenesis, which is consistent with population data linking greater fiber consumption with lowered colorectal cancer (CRC) risk. Butyrate has been shown to regulate the apoptotic response to DNA damage. This study examined the impact of increasing large bowel butyrate concentration by dietary butyrylated starch on the colonic epithelium of rats treated with the genotoxic carcinogen azoxymethane (AOM). Four groups of 10 male rats were fed AIN-93G based-diets containing either low amylose maize starch (LAMS), LAMS with 3% tributyrin, 10% high amylose maize starch (HAMS) or 10% butyrylated HAMS (HAMSB). HAMS and HAMSB starches were cooked by heating in water. After 4 weeks, rats were injected once with AOM and killed 6 h later. Rates of apoptosis and proliferation were measured in colonic epithelium. Short-chain fatty acid concentrations in large bowel digesta and hepatic portal venous plasma were higher in HAMSB than all other groups. Apoptotic rates in the distal colon were increased by HAMSB and correlated with luminal butyrate concentrations but cellular proliferation rates were unaffected by diet. The increase in apoptosis was most marked in the base and proliferative zone of the crypt. Regulation of luminal butyrate using HAMSB increases the rates of apoptotic deletion of DNA-damaged colonocytes. We propose this pro-apoptotic function of butyrate plays a major role reducing tumour formation in the AOM-treated rat and that these data support a potential protective role of butyrate in CRC.


Nonvalidated Home Blood Pressure Devices Dominate the Online Marketplace in Australia: Major Implications for Cardiovascular Risk Management.
Self-home blood pressure (BP) monitoring is recommended to guide clinical decisions on hypertension and is used worldwide for cardiovascular risk management. People usually make their own decisions when purchasing BP devices, which can be made online. If patients purchase nonvalidated devices (those not proven accurate according to internationally accepted standards), hypertension management may be based on inaccurate readings resulting in under- or over-diagnosis or treatment. This study aimed to evaluate the number, type, percentage validated, and cost of home BP devices available online. A search of online businesses selling devices for home BP monitoring was conducted. Multinational companies make worldwide deliveries, so searches were restricted to BP devices available for one nation (Australia) as an example of device availability through the global online marketplace. Validation status of BP devices was determined according to established protocols. Fifty nine online businesses, selling 972 unique BP devices were identified. These included 278 upper-arm cuff devices (18.3% validated), 162 wrist-cuff devices (8.0% validated), and 532 wrist-band wearables (0% validated). Most BP devices (92.4%) were stocked by international e-commerce businesses (eg, eBay, Amazon), but only 5.5% were validated. Validated cuff BP devices were more expensive than nonvalidated devices: median (interquartile range) of 101.1 (75.0-151.5) versus 67.4 (30.4-112.8) Australian Dollars. Nonvalidated BP devices dominate the online marketplace and are sold at lower cost than validated ones, which is a major barrier to accurate home BP monitoring and cardiovascular risk management. Before purchasing a BP device, people should check it has been validated at https://www.stridebp.org.


Physical mapping of a large plant genome using global high-information-content-fingerprinting: the distal region of the wheat ancestor Aegilops tauschii chromosome 3DS.
Physical maps employing libraries of bacterial artificial chromosome (BAC) clones are essential for comparative genomics and sequencing of large and repetitive genomes such as those of the hexaploid bread wheat. The diploid ancestor of the D-genome of hexaploid wheat (Triticum aestivum), Aegilops tauschii, is used as a resource for wheat genomics. The barley diploid genome also provides a good model for the Triticeae and T. aestivum since it is only slightly larger than the ancestor wheat D genome. Gene co-linearity between the grasses can be exploited by extrapolating from rice and Brachypodium distachyon to Ae. tauschii or barley, and then to wheat. We report the use of Ae. tauschii for the construction of the physical map of a large distal region of chromosome arm 3DS. A physical map of 25.4 Mb was constructed by anchoring BAC clones of Ae. tauschii with 85 EST on the Ae. tauschii and barley genetic maps. The 24 contigs were aligned to the rice and B. distachyon genomic sequences and a high density SNP genetic map of barley. As expected, the mapped region is highly collinear to the orthologous chromosome 1 in rice, chromosome 2 in B. distachyon and chromosome 3H in barley. However, the chromosome scale of the comparative maps presented provides new insights into grass genome organization. The disruptions of the Ae. tauschii-rice and Ae. tauschii-Brachypodium syntenies were identical. We observed chromosomal rearrangements between Ae. tauschii and barley. The comparison of Ae. tauschii physical and genetic maps showed that the recombination rate across the region dropped from 2.19 cM/Mb in the distal region to 0.09 cM/Mb in the proximal region. The size of the gaps between contigs was evaluated by comparing the recombination rate along the map with the local recombination rates calculated on single contigs. The physical map reported here is the first physical map using fingerprinting of a complete Triticeae genome. This study demonstrates that global fingerprinting of the large plant genomes is a viable strategy for generating physical maps. Physical maps allow the description of the co-linearity between wheat and grass genomes and provide a powerful tool for positional cloning of new genes.


An easy, inexpensive, and sensitive method for the quantification of chitin in insect peritrophic membrane by image processing.
Chitin, poly (β-(1→4)-N-acetyl-d-glucosamine), is an important biopolymer for insects that is utilized as a major component of peritrophic membrane. The chitin content in peritrophic membrane is of expedient interest from a pest control perspective, although it is hard to quantify chitin. In this study, we establish a facile method for the quantification of chitin in peritrophic membrane by image processing. In this method, chitin was indirectly quantified using chitosan-I3- complex, which exhibited a specific red-purple color. A calibration curve using a chitosan solution showed good linearity in a concentration range of 0.05-0.5 μg/μL. We quantified the amount of chitin in peritrophic membrane of Spodoptera litura (Lepidoptera: Noctuidae) larvae using this method. Throughout the study, only common inexpensive regents and easily attainable apparatuses were employed. This method can be easily applied to the sensitive quantification of the amounts of chitin and chitosan in materials by wide range of researchers. Abbreviations: LOD: limit of detection; LOQ: limit of quantification; ROI: region of interest; RSD: relative standard deviation.


Associations of PrP genotype with lamb production traits in three commercial breeds of British hill sheep.
The National Scrapie Plan (NSP) was launched in Great Britain in 2001, with the aim of eventually eradicating scrapie, a small ruminant transmissible spongiform encephalopathy, from the national sheep flock. Specifically, a selective breeding programme, the Ram Genotyping Scheme, was devised enabling pedigree ram breeders to reduce the number of scrapie-susceptible genotypes from their flocks. The effect of large-scale manipulation of PrP genotypes on commercially important traits within the sheep industry is, however, unknown. We have therefore examined production traits in a total of 43 968 lambs from 32 pedigree breeders across three British hill breeds, comprising 8163 North Country Cheviot (Hill), 21 366 Scottish Blackface and 14 439 Welsh Mountain lambs. Traits examined included: weights at birth, 8 and 20 weeks; ultrasonic fat and muscle depth, and average daily weight gain from 8 to 20 weeks. Linear mixed models were fitted for each trait, including animal (direct) genetic effects and up to three maternal effects. Potential associations with the PrP gene were assessed by fitting either PrP genotype or number of copies of individual alleles as fixed effects. A number of breed-specific significant associations between production traits and the PrP gene were found, but no consistent significant effects were detected across the three breeds. Breed-specific effects were as follows: (i) 0.37 kg higher birth weights (BWTs) in AHQ homozygous North Country Cheviot (Hill) lambs (P < 0.01); (ii) 0.16 kg higher BWTs in ARR homozygous Scottish Blackface lambs (P < 0.05); (iii) 0.5 kg higher 8-week weights in VRQ heterozygous Scottish Blackface lambs (P < 0.01); (iv) a 0.72 kg decrease in scan weight associated with homozygous ARR Welsh Mountain lambs (P < 0.01); (v) 0.51 mm higher ultrasonic muscle depths in AHQ homozygous Welsh Mountain lambs (P < 0.01); (vi) 0.48 mm lower ultrasonic muscle depths in Welsh Mountain lambs carrying one or more copies of the ARR allele (P < 0.05) and (vii) 0.2 mm higher ultrasonic fat depths in heterozygous VRQ Welsh Mountain lambs (P < 0.05). The use of a Bonferroni correction to define appropriate significance thresholds across the three datasets, which account for the large number of independent comparisons made, resulted in breed-specific comparisons, with P < 0.01 becoming significant at P0.05, and the remaining breed-specific comparisons no longer being significant. The absence of a common effect across the three breeds suggests that any true association found may be due to breed-specific alleles of neighbouring genes in linkage disequilibrium with the PrP locus.


Collective Action: The Medical College of Wisconsin COVID-19 Vaccination Program.
The Medical College of Wisconsin (MCW) COVID-19 Vaccination Program facilitated early vaccination efforts in metro Milwaukee, Wisconsin from December, 2020 through April, 2021. Goals of the program were to work with clinical partners to ensure rapid vaccination availability for the institution's frontline workforce, to support state public health agencies in offering a vaccination opportunity for underserved and higher education community members, and to train vaccinators. A key component of the program was the MCW COVID-19 Vaccination Clinic, and 88% of MCW's workforce was fully immunized against COVID-19 with the 2-dose, mRNA vaccine by April 30, 2021. Within the MCW clinic, 219 pharmacy and medical students learned to administer vaccinations, and 12,450 community vaccinations were administered.


Lactobacillus senmaizukei sp. nov., isolated from Japanese pickle.
The taxonomic position of a lactic acid bacterium, strain L13(T), isolated from senmaizuke, a Japanese traditional pickle, was studied. This strain was a heterofermentative, facultatively anaerobic, Gram-positive, non-spore-forming, non-motile, rod-shaped bacterium. It produced dl-lactic acid from glucose. Phylogenetic analysis of its 16S rRNA gene sequence and physiological and biochemical characteristics indicated that the strain was a member of the genus Lactobacillus. Based on 16S rRNA gene sequence similarity, strain L13(T) was closely related to Lactobacillus hammesii CIP 108387(T) (98.9 %) and Lactobacillus parabrevis ATCC 53295(T) (98.5 %). In addition, the rpoA gene sequence of strain L13(T) was closely related to those of L. hammesii CIP 108387(T) (92.2 %) and L. parabrevis ATCC 53295(T) (92.0 %). However, DNA-DNA hybridization of strain L13(T) with these two strains revealed that strain L13(T) represents a separate genomic species. The DNA G+C content of strain L13(T) was 46 mol% and the peptidoglycan was of the A4alpha l-Lys-d-Asp type. Thus, these data indicate that strain L13(T) represents a novel species of the genus Lactobacillus, for which the name Lactobacillus senmaizukei is proposed. The type strain is L13(T) (=NBRC 103853(T)=TISTR 1847(T)).


Herbal medicines for menopausal symptoms.
Many women are now using herbal medicines to try to relieve menopausal symptoms such as hot flushes and night sweats, in light of recent evidence suggesting that hormone replacement therapy (HRT) may increase the likelihood of breast cancer, ovarian cancer, venous thromboembolism, heart attacks and stroke.1-6 For example, one survey has suggested that around 40% of women in the UK have used complementary and alternative treatments for their menopausal symptoms.7 Here we review the efficacy and safety of herbal medicines for the relief of such symptoms.


Stable Chloro- and Bromoxenate Cage Anions; [X3(XeO3)3]3- and [X4(XeO3)4]4- (X = Cl or Br).
The number of isolable compounds which contain different noble-gas-element bonds is limited for xenon and even more so for krypton. Examples of Xe-Cl bonds are rare, and prior to this work, no Xe-Br bonded compound had been isolated in macroscopic quantities. The syntheses, isolation, and characterization of the first compounds to contain Xe-Br bonds and their chlorine analogues are described in the present work. The reactions of XeO3 with [N(CH3)4]Br and [N(C2H5)4]Br have provided two bromoxenate salts, [N(C2H5)4]3[Br3(XeO3)3] and [N(CH3)4]4[Br4(XeO3)4], in which the cage anions have Xe-Br bond lengths that range from 3.0838(3) to 3.3181(8) Å. The isostructural chloroxenate anions (Xe-Cl bond lengths, 2.9316(2) to 3.101(4) Å) were synthesized by analogy with their bromine analogues. The bromo- and chloroxenate salts are stable in the atmosphere at room temperature and were characterized in the solid state by Raman spectroscopy and low-temperature single-crystal X-ray diffraction, and in the gas phase by quantum-chemical calculations. They are the only known examples of cage anions that contain a noble-gas element. The Xe-Br and Xe-Cl bonds are very weakly covalent and can be viewed as σ-hole interactions, similar to those encountered in halogen bonding. However, the halogen atoms in these cases are valence electron lone pair donors, and the σ*Xe-O orbitals are lone pair acceptors.


Rational design of humanized antibody inhibitors for cathepsin S.
Cathepsin S (CTSS) is involved in pathogenesis of many human diseases. Inhibitors blocking its protease activity hold therapeutic potential. In comparison to small-molecule inhibitors, monoclonal antibodies capable of inhibiting CTSS enzymatic activity may possess advantageous pharmacological properties. Here we designed and produced inhibitory antibodies targeting human CTSS by genetically fusing the propeptide of procathepsin S (proCTSS) with antibodies in clinic. The resulting antibody fusions in full-length or fragment antigen-binding format could be stably expressed and potently inhibit CTSS proteolytic activity in high specificity. These fusion antibodies not only demonstrate a new approach for facile synthesis of antibody inhibitors against CTSS, but also represent novel anti-CTSS therapeutic candidates.


What the Eyes Hear: An Eye-Tracking Study on Phonological Awareness in Emirati Arabic.
Phonological awareness is the ability to perceive and manipulate the sounds of spoken words. It is considered a good predictor of reading and spelling abilities. In the current study, we used an eye-tracking procedure to measure fixation differences while adults completed three conditions of phonological awareness in Emirati Arabic (EA): (1) explicit instructions for onset consonant matching (OCM), (2) implicit instructions for segmentation of initial consonant (SIC), and (3) rhyme matching (RM). We hypothesized that fixation indices would vary according to the experimental conditions. We expected explicit instructions to facilitate task performance. Thus, eye movements should reflect more efficient fixation patterns in the explicit OCM condition in comparison to the implicit SIC condition. Moreover, since Arabic is consonant-based, we hypothesized that participants would perform better in the consonant conditions (i.e., OCM and SIC) than in the rhyme condition (i.e., RM). Finally, we expected that providing feedback during practice trials would facilitate participants' performance overall. Response accuracy, expressed as a percentage of correct responses, was recorded alongside eye movement data. Results show that performance was significantly compromised in the RM condition, where targets received more fixations of longer average duration, and significantly longer gaze durations in comparison to the OCM and SIC conditions. Response accuracy was also significantly lower in the RM condition. Our results indicate that eye-tracking can be used as a tool to test phonological awareness skills and shows differences in performance between tasks containing a vowel or consonant manipulation.


Coral skeletons defend against ultraviolet radiation.
Many coral reef organisms are photosynthetic or have evolved in tight symbiosis with photosynthetic symbionts. As such, the tissues of reef organisms are often exposed to intense solar radiation in clear tropical waters and have adapted to trap and harness photosynthetically active radiation (PAR). High levels of ultraviolet radiation (UVR) associated with sunlight, however, represent a potential problem in terms of tissue damage. By measuring UVR and PAR reflectance from intact and ground bare coral skeletons we show that the property of calcium carbonate skeletons to absorb downwelling UVR to a significant extent, while reflecting PAR back to the overlying tissue, has biological advantages. We placed cnidarians on top of bare skeletons and a UVR reflective substrate and showed that under ambient UVR levels, UVR transmitted through the tissues of cnidarians placed on top of bare skeletons were four times lower compared to their counterparts placed on a UVR reflective white substrate. In accordance with the lower levels of UVR measured in cnidarians on top of coral skeletons, a similar drop in UVR damage to their DNA was detected. The skeletons emitted absorbed UVR as yellow fluorescence, which allows for safe dissipation of the otherwise harmful radiation. Our study presents a novel defensive role for coral skeletons and reveals that the strong UVR absorbance by the skeleton can contribute to the ability of corals, and potentially other calcifiers, to thrive under UVR levels that are detrimental to most marine life.


Stochastic thermodynamics with arbitrary interventions.
We extend the theory of stochastic thermodynamics in three directions: (i) instead of a continuously monitored system we consider measurements only at an arbitrary set of discrete times, (ii) we allow for imperfect measurements and incomplete information in the description, and (iii) we treat arbitrary manipulations (e.g., feedback control operations) which are allowed to depend on the entire measurement record. For this purpose we define for a driven system in contact with a single heat bath the four key thermodynamic quantities-internal energy, heat, work, and entropy-along a single "trajectory" for a causal model. The first law at the trajectory level and the second law on average is verified. We highlight the special case of Bayesian or "bare" measurements (incomplete information, but no average disturbance) which allows us to compare our theory with the literature and to derive a general inequality for the estimated free energy difference in Jarzynski-type experiments. An analysis of a recent Maxwell demon experiment using real-time feedback control is also given. As a mathematical tool, we prove a classical version of Stinespring's dilation theorem, which might be of independent interest.


Alimentary-associated carcinomas in five Vietnamese potbellied pigs.
5 Vietnamese potbellied pigs were evaluated for abdominal distress that had not responded to medical treatment (4 pigs) or a draining tract of the cranial abdomen of unknown duration (1 pig). Clinical signs in the pigs included anorexia, vomiting, and constipation. Physical examination revealed a palpable abdominal mass in all pigs. Radiography revealed distended loops of small intestine in 2 pigs. 3 pigs were treated successfully with wide-margin excision of the abdominal masses, and 2 were euthanized. Primary tumors were diagnosed at necropsy or through histologic evaluation of biopsy specimens obtained during surgery. Types of tumor included cholangiocellular carcinoma, transmural gastric carcinoma, small intestinal adenocarcinoma, metastatic hepatocellular carcinoma, and carcinoma. The tumors involved the stomach, small intestine, spiral colon, liver, and gall bladder. All 3 surgically treated pigs survived at least 9 months after surgery. Although rare, neoplasia of the alimentary system should be considered among the differential diagnoses for potbellied pigs with signs of abdominal distress. Wide-margin excision of the neoplastic tissue may result in a good outcome in affected pigs.


Integrate imaging approach for minimally invasive and robotic procedures.
Over the past two decades, robotic and minimally invasive cardiac surgery has been continuously refined and is currently an alternative to traditional open-heart surgery for some patients. The parallel evolution of imaging modalities has made robotic surgery safer and more efficient. Here, we review the pre- and post-operative use of computed tomography (CT) in minimally invasive and robotic cardiac procedures.


Role of heterotrimeric G proteins in platelet activation and clot formation in platelets treated with integrin αIIbβ3 inhibitor.
Mechanisms of platelet activation are triggered by thrombin, adenosine diphosphate (ADP), epinephrine, thromboxane A2, and other soluble agonists which induce signaling via heterotrimeric Gαq, Gαi, and Gα12/13 proteins. We have undertaken a study addressing the contribution of these G proteins to platelet activation and clot formation in the presence of eptifibatide, thus excluding outside-in signaling provided by integrin αIIbβ3-fibrinogen engagement. Selective and combined activation of the G proteins was achieved by using combinations of platelet agonists and inhibitors. Platelet activation in platelet-rich plasma was evaluated by P-selectin expression using flow cytometry. Contribution of platelets to whole blood clotting was assessed by rotation thromboelastometry (ROTEM). Selective signaling of Gαq or Gαi but not Gα12/13 promoted P-selectin expression. Further enhancement of P-selectin expression was achieved by ADP-induced combined signaling of Gαq and Gαi, and to more extent by U46619 at high concentration (1.5 μM) induced combined signaling of Gαq and Gα12/13 while maximal P-selectin expression was achieved by thrombin receptor-activating peptide (TRAP)-induced combined signaling of Gαq, Gαi, and Gα12/13. In ROTEM, selective activation of Gαq, Gαi, or Gα12/13 failed to affect blood clotting. Combined signaling of Gαq and Gαi or Gαq and Gα12/13 or all three G proteins shortened the clotting time and stimulated clot strength. Pretreatment of platelets with acetylsalicylic acid did not change the effect of ADP but inhibited the effect of TRAP. Signaling of Gαq and Gα12/13 triggered by U46619 also stimulated clot formation. Combined signaling of either Gαq and Gαi or Gαq and Gα12/13 is sufficient to stimulate maximal platelet activation and enhanced clot formation in platelets treated with inhibitor of integrin αIIbβ3. It could be suggested that outside-in signaling is not necessarily required to fulfill these platelet functions.


Nonadditive genetic effects in animal behavior.
Heritabilities, commonly used to predict evolutionary potential, are notoriously low for behaviors. Apart from strong contributions of environmental variance in reducing heritabilities, the additive genetic components can be very low, especially when they are camouflaged by nonadditive genetic effects. We first report the heritabilities of courtship traits in founder-flush and control populations of the housefly (Musca domestica L.). We estimated the heritability of each male and female display through the regression of the courtships involving daughters and sons (with randomly selected mates) onto the "midparental" courtship values of their parents. Overall, the average heritability was significantly (P = .012) higher for the parent-daughter assays than for the parent-son assays. We attributed the low (even negative) heritabilities to genotype-by-environment interactions whereby the male's behavior is influenced by the "environment" of his mating partner's preferences for the display, generating epistasis through indirect genetic effects. Moreover, bottlenecked lines had up to 800% of the heritability of the controls, suggesting "conversion" of additive genetic variance from nonadditive components. Second, we used line-cross assays on separate populations that had been selected for divergence in mating behavior to identify dominance and epistasis through heterosis and outbreeding depression in courtship. Finally, our literature review confirms the prevalence of such low heritabilities (i.e., a conservative mean of 0.38) and nonadditive genetics in other behavioral repertoires (64% of the studies). We conclude that animal behavior is especially prone to the gamut of quantitative genetic complexities that can result in negative heritabilities, negative selection responses, inbreeding depression, conversion, heterosis, and outbreeding depression.


Plant sterols as dietary adjuvants in the reduction of cardiovascular risk: theory and evidence.
Plant sterol-enriched foods are an effective dietary adjuvant in reducing cardiovascular risk by lowering total cholesterol and low density lipoprotein-cholesterol (LDL-C) in serum by up to approximately 15%. The mechanism of action of plant sterols is different from those of 3-hydroxy-3-methylglutaryl coenzyme A inhibitors (statins) and thus their effect is additive. Combining plant sterols with other dietary components known to reduce cholesterol in a portfolio approach has proven to be most effective for reduction of hypercholesterolemia and provide an alternative treatment option for clinicians. Plant sterol-enriched foods provides clinicians with a relatively cheap, safe, and effective way to help patients manage their cardiovascular risk.


Exploiting the proteome to improve the genome-wide genetic analysis of epistasis in common human diseases.
One of the central goals of human genetics is the identification of loci with alleles or genotypes that confer increased susceptibility. The availability of dense maps of single-nucleotide polymorphisms (SNPs) along with high-throughput genotyping technologies has set the stage for routine genome-wide association studies that are expected to significantly improve our ability to identify susceptibility loci. Before this promise can be realized, there are some significant challenges that need to be addressed. We address here the challenge of detecting epistasis or gene-gene interactions in genome-wide association studies. Discovering epistatic interactions in high dimensional datasets remains a challenge due to the computational complexity resulting from the analysis of all possible combinations of SNPs. One potential way to overcome the computational burden of a genome-wide epistasis analysis would be to devise a logical way to prioritize the many SNPs in a dataset so that the data may be analyzed more efficiently and yet still retain important biological information. One of the strongest demonstrations of the functional relationship between genes is protein-protein interaction. Thus, it is plausible that the expert knowledge extracted from protein interaction databases may allow for a more efficient analysis of genome-wide studies as well as facilitate the biological interpretation of the data. In this review we will discuss the challenges of detecting epistasis in genome-wide genetic studies and the means by which we propose to apply expert knowledge extracted from protein interaction databases to facilitate this process. We explore some of the fundamentals of protein interactions and the databases that are publicly available.


Treatment and stabilization of complex wounds involving the pelvic bone, groin, and femur with the inferiorly based rectus abdominis musculocutaneous flap and the use of power color Doppler imaging in preoperative evaluation.
The authors present case reports demonstrating the trilevel utility of the inferiorly based rectus abdominis musculocutaneous flap in the closure of complex wounds involving the pelvis, groin, and femur that had failed previously or were not amenable to traditional closure techniques. The use of the rectus abdominis flap was especially advantageous for achieving infection eradication and large dead space closure. Additionally they present the emerging technique of power color Doppler imaging as a valuable tool in preoperative flap planning. This technique is particularly useful in evaluating the candidacy for rectus abdominis musculocutaneous flap placement of patients with a prior history of abdominal surgeries, trauma, infection, irradiation, or other conditions that might compromise the patency of the deep inferior epigastric vessels.


The inhibitory effects of nano-Ag on voltage-gated potassium currents of hippocampal CA1 neurons.
The application of the nano-sized materials continues to grow at a rapid rate in the fields of medicine, biotechnology, and environmental technology. Voltage-gated potassium currents play a key role in excitable cellular viability and function, especially in the central nervous system. The aim of this study was to investigate the actions of silver nano-particles (nano-Ag) on voltage-activated potassium currents in hippocampal CA1 neurons using whole cell patch-clamp technique. The hydrodynamic mean diameter of nano-Ag (10(-5) g mL(-1) ) was 223.9 nm in artificial cerebrospinal fluid (ACSF). Both types, transient potassium (I(A) ) and delayed rectifier potassium (I(K) ) current amplitudes were inhibited by the nano-Ag (10(-5) g mL(-1) ). The nano-Ag particles produced a hyperpolarizing shift in the activation-voltage curve of I(K) and inactivation-voltage curve of I(A) and also delayed the recovery of I(A) from inactivation. The results suggest that nano-Ag may have potential to alter the excitability of neurons by depressing the potassium channels.


AGROBACTERIUM TUMEFACIENS: THERMAL INACTIVATION OF TUMOR-INDUCING ABILITY.
Treating cultures of Agrobacterium tumefaciens at 39 degrees to 48 degrees C reduces the infectivity of the bacteria without necessarily affecting viability. Destruction of the capacity to initiate tumor growth follows first-order kinetics from which rate constants for thermal inactivation are derived. From these rates, values for heat of activation of 56.7 kcal mole(-1) and entropy of activation of 107 cal mole(-1) deg(-1) are obtained. A particular protein or nucleoprotein active in the process of infection may be inactivated by the treatment.


Double standards: the multinational asbestos industry and asbestos-related disease in South Africa.
This study documents and contrasts the development of knowledge about asbestos-related disease (ARD) in South Africa and the United Kingdom. It also contributes to the globalization debate by exploring corporate decision-making in a multinational industry. Between the 1930s and 1960s, the leading U.K. asbestos companies developed a sophisticated knowledge of ARD, though in South Africa, where the leading companies such as Turner & Newall and Cape Asbestos owned mines, there was little attempt to apply this knowledge. Asbestos mines (and their environments) in South Africa were uniquely dusty and ARD was rife. Social and political factors in South Africa, especially apartheid, allowed these companies to apply double standards, even after 1960 when the much more serious hazard of mesothelioma was identified. This shows the need for greater regulation of multinationals. Because of the lack of such regulation in the early 1960s, an opportunity was lost to prevent the current high morbidity and mortality of ARD both in South Africa and worldwide.


Determinations of the DNA sequence of the mreB gene and of the gene products of the mre region that function in formation of the rod shape of Escherichia coli cells.
The 6.5-kilobase mre region at 71 min in the Escherichia coli chromosome map, where genes involved in formation of a rod-shaped cell form a gene cluster, was analyzed by in vivo protein synthesis in a maxicell system and by base sequencing of DNA. An open reading frame that may code for a protein with an Mr of about 37,000 on sodium dodecyl sulfate-polyacrylamide gels was found and was correlated with the mreB gene. N-terminal amino acid sequencing of the hybrid mreB-lacZ protein confirmed the production by mreB of a protein of 347 amino acid residues with a molecular weight of 36,958. The amino acid sequence of this protein deduced from the DNA sequence showed close similarity with that of a protein of the ftsA gene which is involved in cell division of E. coli. Three other contiguous genes that formed three proteins with Mrs of about 40,000, 22,000, and 51,000, respectively, were detected downstream of the mreB gene by in vivo protein synthesis. The mreB protein and some of these three proteins may function together in determination of cell shape.


Reproducibility of mental stress-induced myocardial ischemia in the Psychophysiological Investigations of Myocardial Ischemia (PIMI).
Many patients with coronary artery disease (CAD) develop myocardial ischemia in response to mental stress. This has been documented both in the natural environment and in the laboratory. However, the reproducibility of laboratory mental stress-induced ischemia has not been investigated. Sixty patients with documented CAD and a positive exercise stress test discontinued cardiac medications and underwent two standardized mental stress tests (a timed Stroop Color-Word test and a public speaking task) in a nuclear cardiology laboratory (Visit 1), and repeated this procedure between 2 and 8 weeks later (Visit 2). Measurements of cardiovascular function and neurohormonal responses were obtained throughout testing, and mood state was assessed before and after testing. Sixty-eight percent of the 56 patients with detailed radionuclide data from both visits had consistent responses (ie, ischemia either present during both sessions or absent during both) to the Stroop task (kappa = .29, p = .03), 61% had consistent responses to the speech task (kappa = .20, p = .12), and 60% had consistent responses when ischemia was considered present if it occurred during either the Stroop test, the speech task, or both, and absent if it did not occur during either task (kappa = .22, p = .07). Hemodynamic and neuroendocrine responses to the tests were moderately reproducible. We conclude that two popular laboratory tests for mental stress-induced myocardial ischemia are modestly reproducible. The relatively low reproducibility is probably influenced by uncertainties in detecting relatively small changes in wall motion, habituation of the patient to repeated exposure to psychological stressors, and physiological differences in threshold for ischemia on different days of testing.


Rationalization and prediction of drug resistant mutations in targets for clinical anti-tubercular drugs.
Resistance to therapy limits the effectiveness of drug treatment in many diseases. Drug resistance can be considered as a successful outcome of the bacterial struggle to survive in the hostile environment of a drug-exposed cell. An important mechanism by which bacteria acquire drug resistance is through mutations in the drug target. Drug resistant strains (multi-drug resistant and extensively drug resistant) of Mycobacterium tuberculosis are being identified at alarming rates, increasing the global burden of tuberculosis. An understanding of the nature of mutations in different drug targets and how they achieve resistance is therefore important. An objective of this study is to first decipher sequence as well as structural bases for the observed resistance in known drug resistant mutants and then to predict positions in each target that are more prone to acquiring drug resistant mutations. A curated database containing hundreds of mutations in the 38 drug targets of nine major clinical drugs, associated with resistance is studied here. Mutations have been classified into those that occur in the binding site itself, those that occur in residues interacting with the binding site and those that occur in outer zones. Structural models of the wild type and mutant forms of the target proteins have been analysed to seek explanations for reduction in drug binding. Stability analysis of an entire array of 19 mutations at each of the residues for each target has been computed using structural models. Conservation indices of individual residues, binding sites and whole proteins are computed based on sequence conservation analysis of the target proteins. The analyses lead to insights about which positions in the polypeptide chain have a higher propensity to acquire drug resistant mutations. Thus critical insights can be obtained about the effect of mutations on drug binding, in terms of which amino acid positions and therefore which interactions should not be heavily relied upon, which in turn can be translated into guidelines for modifying the existing drugs as well as for designing new drugs. The methodology can serve as a general framework to study drug resistant mutants in other micro-organisms as well.


CPAP Treatment Adherence in Women with Obstructive Sleep Apnea.
Untreated obstructive sleep apnea (OSA) has numerous negative health-related consequences. Continuous positive airway pressure (CPAP) is generally considered the treatment of choice for OSA, but rates of nonadherence are high. It is believed that OSA is more prevalent among men; therefore understanding how OSA presents among women is limited and treatment adherence has received little research attention. For this study, 29 women were recruited from primary care offices. They completed a questionnaire battery and underwent a night of nocturnal polysomnography (PSG) followed by a visit with a sleep specialist. Women diagnosed with OSA were prescribed CPAP; 2 years later CPAP adherence was evaluated. Results show that approximately half the sample was adherent. There were no significant differences between adherent and nonadherent women on OSA severity; however CPAP adherent women had worse nocturnal and daytime functioning scores at the time of diagnosis. Moreover, when the seven nocturnal and daytime variables were used as predictors in a discriminant analysis, they could predict 87% of adherent and 93% of the nonadherent women. The single most important predictor was nonrefreshing sleep. We discuss the implications of the findings for identifying women in primary care with potential OSA and offer suggestions for enhancing treatment adherence.


Electronic data collection by trainee anaesthetists using palm top computers.
The Royal College of Anaesthetist requires that trainees wishing to specialise in Anaesthesia register with the College and keep a logbook documenting their experience and training in clinical anaesthesia. There are a number of advantages if such logbooks are kept in an electronic format. For the first time in the United Kingdom, we have evaluated the usefulness in a district general hospital of introducing a computerized system of record keeping based on a portable palm top computer. This has enabled the careful analysis of the individual work load of each trainee in the department by means of a specific program designed to generate a comprehensive report after the information from each individual has been downloaded on to a standard desk top computer. We have found a number of interesting and unexpected results and have highlighted ways in which the system may be improved.


Revealing enzyme functional architecture via high-throughput microfluidic enzyme kinetics.
Systematic and extensive investigation of enzymes is needed to understand their extraordinary efficiency and meet current challenges in medicine and engineering. We present HT-MEK (High-Throughput Microfluidic Enzyme Kinetics), a microfluidic platform for high-throughput expression, purification, and characterization of more than 1500 enzyme variants per experiment. For 1036 mutants of the alkaline phosphatase PafA (phosphate-irrepressible alkaline phosphatase of Flavobacterium), we performed more than 670,000 reactions and determined more than 5000 kinetic and physical constants for multiple substrates and inhibitors. We uncovered extensive kinetic partitioning to a misfolded state and isolated catalytic effects, revealing spatially contiguous regions of residues linked to particular aspects of function. Regions included active-site proximal residues but extended to the enzyme surface, providing a map of underlying architecture not possible to derive from existing approaches. HT-MEK has applications that range from understanding molecular mechanisms to medicine, engineering, and design.


Silicic acid limitation drives bloom termination and potential carbon sequestration in an Arctic bloom.
The spring diatom bloom in the Arctic Ocean accounts for significant annual primary production leading to the most rapid annual drawdown of water-column pCO2. Late-winter waters in the Atlantic Arctic & Subarctic Provinces (AASP) have lower silicic acid concentrations than nitrate, which suggests diatom blooms may deplete Si before N. Here we test a facet of the hypothesis that silicic acid limitation terminates the spring diatom bloom in the AASP and the sinking of the senescent and dead diatoms helps drive carbon sequestration. During a 6-week study, diatoms bloomed and progressively consumed silicic acid to where it limited their growth. The onset of growth limitation was concurrent with the minimum pCO2 in the surface waters and increases in both the proportion of dead diatoms and the diatom assemblage sedimentation rate. Data reanalysis within the AASP shows a highly significant and positive correlation between silicic acid and pCO2 in the surface waters, but no significant relationship with nitrate and pCO2 was observed unless data were smoothed. Therefore, understanding the future of the AASP spring diatom bloom requires models that explicitly consider changes in silicic acid supply as a driver of this process.


[Cyclosporin induced gingival hyperplasia].
Cyclosporine is a strong immunosuppressive drug, which is used specially in transplantation patients, however in some cases, like others medicines it causes, gingival overgrowth. In the present article we present five transplantations patients, (one of liver, and four of kidney), whom have taken this drug since more than a year at different dosages. One of them, never has presented overgrowth, whereas the rest cases have needed this specific periodontal treatment to solve hyperplasy. After this work, we don't find clear relationship between drug dosage-age-effect over tissues.


Association of WHSC1/NSD2 and T-cell infiltration with prostate cancer metastasis and prognosis.
Progress in immunotherapy for prostate cancer (PCa) lags that for other cancers, mainly because of limited immune infiltration in PCa. This study aimed to assess the feasibility of NSD2 as an immunotherapeutic target in PCa. Immunohistochemistry was performed to evaluate the expression pattern of NSD2 in 34 cases of benign prostatic hyperplasia (BPH), 36 cases of prostatic intraepithelial neoplasia (PIN), and 57 cases of PCa, including 19 cases of metastatic castration-resistant prostatic cancer (mCRPC). Single-cell RNA sequencing and gene set enrichment analysis (GSEA) were used to correlate NSD2 with certain downstream pathways. Furthermore, the Immuno-Oncology-Biological-Research (IOBR) software package was used to analyze the potential roles of NSD2 in the tumor microenvironment. We found that the positive expression rate of NSD2 increased progressively in BPH, PIN and PCa. mCRPC had the highest staining intensity for NSD2. High NSD2 expression was positively correlated with the infiltration level of CD4+ tumor-infiltrating lymphocytes (TILs) and negatively correlated with that of CD8+ TILs. Importantly, a new immune classification based on NSD2 expression and CD4+ TILs and CD8+ TILs was successfully used to stratify PCa patients based on OS.PSA and CD4+ TILs are independent risk factors for PCa bone metastasis. This study demonstrates a novel role for NSD2 in defining immune infiltrate on in PCa and highlights the great potential for its application in immunotherapy response evaluation for prostate malignancies.


Oxidative stress in IgA nephropathy.
IgA nephropathy (IgAN) is characterized by mesangial deposits of IgA1, likely due to accumulation of IgA immune complexes. The activation of intracellular signaling mostly results in oxidative stress, as detected in mesangial cells cultured with aberrantly glycosylated IgA or IgA aggregates and in renal biopsies of patients with IgAN. Signs of altered oxidation/antioxidation balance have been detected in sera and/or in erythrocytes of patients with IgAN, including increased levels of lipoperoxide or malondialdehyde and reduced activity of superoxide dismutase, catalase and glutathione peroxidase. Moreover, increased levels of a marker of oxidative stress, advanced oxidation protein products (AOPPs), have been reported to be significantly associated with proteinuria and disease progression in patients with IgAN. AOPPs are often carried by albumin and can in turn enhance the oxidative stress in the circulation. Recent research suggests that the nephrotoxicity of aberrantly glycosylated IgA1 in IgAN is enhanced in the presence of systemic signs of oxidative stress, and it is tempting to hypothesize that the level of the oxidative milieu conditions the different expression and progression of IgAN.


Inter-viral conflicts that exploit host CRISPR immune systems of Sulfolobus.
Infection of Sulfolobus islandicus REY15A with mixtures of different Sulfolobus viruses, including STSV2, did not induce spacer acquisition by the host CRISPR immune system. However, coinfection with the tailed fusiform viruses SMV1 and STSV2 generated hyperactive spacer acquisition in both CRISPR loci, exclusively from STSV2, with the resultant loss of STSV2 but not SMV1. SMV1 was shown to activate adaptation while itself being resistant to CRISPR-mediated adaptation and DNA interference. Exceptionally, a single clone S-1 isolated from an SMV1 + STSV2-infected culture, that carried STSV2-specific spacers and had lost STSV2 but not SMV1, acquired spacers from SMV1. This effect was also reproducible on reinfecting wild-type host cells with a variant SMV1 isolated from the S-1 culture. The SMV1 variant lacked a virion protein ORF114 that was shown to bind DNA. This study also provided evidence for: (i) limits on the maximum sizes of CRISPR loci; (ii) spacer uptake strongly retarding growth of infected cultures; (iii) protospacer selection being essentially random and non-directional, and (iv) the reversible uptake of spacers from STSV2 and SMV1. A hypothesis is presented to explain the interactive conflicts between SMV1 and the host CRISPR immune system.


Angiogenesis and blood vessel stability in inflammatory arthritis.
To assess blood vessel stability in inflammatory synovial tissue (ST) and to examine neural cell adhesion molecule (NCAM), oxidative DNA damage, and hypoxia in vivo. Macroscopic vascularity and ST oxygen levels were determined in vivo in patients with inflammatory arthritis who were undergoing arthroscopy. Vessel maturity/stability was quantified in matched ST samples by dual immunofluorescence staining for factor VIII (FVIII)/alpha-smooth muscle actin (alpha-SMA). NCAM and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) were examined by immunohistochemistry. Angiogenesis was assessed in vitro, using human dermal endothelial cells (HDECs) in a Matrigel tube formation assay. A significant number of immature vessels (showing no pericyte recruitment) was observed in tissue from patients with inflammatory arthritis (P < 0.001), in contrast to osteoarthritic and normal tissue, which showed complete recruitment of pericytes. Low in vivo PO(2) levels in the inflamed joint (median [range] 22.8 [3.2-54.1] mm Hg) were inversely related to increased macroscopic vascularity (P < 0.04) and increased microscopic expression of FVIII and alpha-SMA (P < 0.04 and P < 0.03, respectively). A significant proportion of vessels showed focal expression of NCAM and strong nuclear 8-oxodG expression, implicating a loss of EC-pericyte contact and increased DNA damage, levels of which were inversely associated with low in vivo PO(2) (P = 0.04 for each comparison). Circulating cells were completely negative for 8-oxodG. Exposure of HDEC to 3% O(2) (reflecting mean ST in vivo measurements) significantly increased EC tube formation (P < 0.05). Our findings indicate the presence of unstable vessels in inflamed joints associated with hypoxia, incomplete EC-pericyte interactions, and increased DNA damage. These changes may further contribute to persistent hypoxia in the inflamed joint to further drive this unstable microenvironment.


In situ metagenomics: A platform for rapid sequencing and analysis of metagenomes in less than one day.
We present here a complete system for metagenomic analysis that allows performing the sequencing and analysis of a medium-size metagenome in less than one day. This unprecedented development was possible due to the conjunction of state-of-the-art experimental and computational advances: a portable laboratory suitable for DNA extraction and sequencing with nanopore technology; the powerful metagenomic analysis pipeline SqueezeMeta, capable to provide a complete analysis in a few hours and using scarce computational resources; and tools for the automatic inspection of the results via a graphical user interface, that can be coupled to a web server to allow remote visualization of data (SQMtools and SQMxplore). We have tested the feasibility of our approach in the sequencing of the microbiota associated to volcanic rocks in La Palma, Canary Islands. Also, we did a two-day sampling campaign of marine waters in which the results obtained on the first day guided the experimental design of the second day. We demonstrate that it is possible to generate metagenomic information in less than one day, making it feasible to obtain taxonomic and functional profiles fast and efficiently, even in field conditions. This capacity can be used in the further to perform real-time functional and taxonomic monitoring of microbial communities in remote areas.


Large fluctuations in speed on Greenland's Jakobshavn Isbrae glacier.
It is important to understand recent changes in the velocity of Greenland glaciers because the mass balance of the Greenland Ice Sheet is partly determined by the flow rates of these outlets. Jakobshavn Isbrae is Greenland's largest outlet glacier, draining about 6.5 per cent of the ice-sheet area, and it has been surveyed repeatedly since 1991 (ref. 2). Here we use remote sensing data to measure the velocity of Jakobshavn Isbrae between 1992 and 2003. We detect large variability of the velocity over time, including a slowing down from 6,700 m yr(-1) in 1985 to 5,700 m yr(-1) in 1992, and a subsequent speeding up to 9,400 m yr(-1) by 2000 and 12,600 m yr(-1) in 2003. These changes are consistent with earlier evidence for thickening of the glacier in the early 1990s and rapid thinning thereafter. Our observations indicate that fast-flowing glaciers can significantly alter ice discharge at sub-decadal timescales, with at least a potential to respond rapidly to a changing climate.


[Experimental development of the principles for designing an optimal schedule for inoculations with an inactivated cultured rabies vaccine].
Immune response was studied experimentally in mice immunized according to 23 different schedules with rabies vaccine in 1:10 and 1:20 dilutions for the elucidation of principles of developing an optimal vaccination schedule. Each group consisted of 40 to 50 animals. The most objective, reliable, and strict test, challenge of immunity to street rabies virus inoculated peripherally which is closest to natural conditions, was used as the criterion for the evaluation of immune response. The studies showed daily inoculation of the vaccine not to be the optimal schedule for the manifestation of the immune response after vaccinations against rabies. Most intensive immunity was observed after vaccination with intervals between the injections (principle 1). When the vaccine was inoculated at a 5-10-day interval, a booster effect of the type of secondary immune response was observed, and the protection of the animals was significantly higher (p less than 0.001) than after daily inoculation of the same amount of the vaccine, significantly higher than after daily inoculation of the double amount of the vaccine, and as high as after inoculation of the 4-fold amount of the vaccine daily for 8 days. No booster effect of the vaccine was observed when the interval between the injections was no more than 2-3 days. The booster effect of the vaccine injections at intervals is determined by the level of the ground immunity (priming immunization) (principle 2) and the antigenic potency of the vaccine (principle 3).


Dumping and illegal transport of hazardous waste, danger of modern society.
Increasing the production of hazardous waste during the past few years and stricter legislation in the area of permanent disposal and transportation costs were significantly elevated above activities. This creates a new, highly lucrative gray market which opens the way for the criminalization. Of great importance is the identification of illegal trafficking of hazardous waste since it can have a significant impact on human health and environmental pollution. Barriers to effective engagement to prevent these activities may vary from region to region, country to country, but together affect the ability of law enforcement authorities to ensure that international shipments of hazardous waste comply with national laws and maritime regulations. This paper will overview the legislation governing these issues, and to analyze the barriers to their implementation, but also try to answer the question of why and how this type of waste traded. Paper is an overview of how Croatia is prepared to join the European Union in this area and indicates the importance and necessity of the cooperation of all of society, and international organizations in the fight with the new trend of environmental crime.


Allele frequency of resistance to Bacillus thuringiensis Cry1ab corn in Louisiana populations of sugarcane borer (Lepidoptera: Crambidae).
Transgenic Bacillus thuringiensis (Bt) corn, Zea mays L., has been widely used to manage a corn borer complex in the mid-southern region of the United States. The sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), has become a dominant cornstalk boring species in some areas of this region, especially in Louisiana. Therefore, management of sugarcane borer resistance to Bt corn is critical to ensure the long-term sustainability of Bt corn for the region. This study screened 280 two-parent family-lines of sugarcane borer from four geographical populations in Louisiana during 2005 to determine whether Bt resistance allele frequency in sugarcane borer is sufficiently low to meet the rare resistance assumption of the current "high dose/refuge" resistance management strategy for Bt corn. These sugarcane borer family-lines were examined for Bt resistance by using novel F2 screening procedures. No major Bt resistance alleles were detected in these four populations. The estimated frequency of major Bt resistance alleles was < 0.0027, with a 95% probability and a detection power of 94%. The estimated minor resistance allele frequency was 0.0063, with a 95% CI of 0.0025-0.0117. During a previous study, a major Bt resistance allele was detected in one individual from 213 family-lines of another Louisiana population of sugarcane borer. Combining these data with the current screen, the frequency of major Bt resistance alleles across the five populations was 0.001, with a 95% credibility interval of 0.0001-0.0028 and a detection power of 95%. Major Bt resistance allele frequencies in Louisiana sugarcane borer populations seem to be low, and they should support the rare resistance allele requirement of the high dose/refuge strategy.


Reductions in employee productivity impairment observed after implementation of web-based worksite health promotion programs.
To assess changes in employee productivity impairment observed after the implementation of several Web-based health promotion programs. Health risk assessments and self-report measures of productivity impairment were administered on-line to more than 43,000 participants of Web-based health promotion programs. Reductions in productivity impairment were observed after 1 month of program utilization. Productivity impairment at 90- and 180-day follow-ups also decreased relative to baseline. Improvements in employee health were associated with reductions in employee productivity impairment. The use of Web-based health promotion programs was associated with reductions in productivity impairment and improvements in employee health. After the implementation of Web-based health promotion programs, reductions in productivity impairment may be observed before reductions in direct health care costs.


Reward circuitry function in autism spectrum disorders.
Social interaction deficits and restricted repetitive behaviors and interests that characterize autism spectrum disorders (ASDs) may both reflect aberrant functioning of brain reward circuits. However, no neuroimaging study to date has investigated the integrity of reward circuits using an incentive delay paradigm in individuals with ASDs. In the present study, we used functional magnetic resonance imaging to assess blood-oxygen level-dependent activation during reward anticipation and outcomes in 15 participants with an ASD and 16 matched control participants. Brain activation was assessed during anticipation of and in response to monetary incentives and object image incentives previously shown to be visually salient for individuals with ASDs (e.g., trains, electronics). Participants with ASDs showed decreased nucleus accumbens activation during monetary anticipation and outcomes, but not during object anticipation or outcomes. Group × reward-type-interaction tests revealed robust interaction effects in bilateral nucleus accumbens during reward anticipation and in ventromedial prefrontal cortex during reward outcomes, indicating differential responses contingent on reward type in these regions. Results suggest that ASDs are characterized by reward-circuitry hypoactivation in response to monetary incentives but not in response to autism-relevant object images. The clinical implications of the double dissociation of reward type and temporal phase in reward circuitry function in ASD are discussed.


Proteomic analysis of a developmentally regulated secretory vesicle.
Secretion of spore coat proteins from the prespore secretory vesicles (PSVs) in Dictyostelium discoideum is a signal mediated event that underlies terminal cell differentiation, and represents an important case of developmentally regulated secretion. In order to study the biochemical mechanisms that govern the regulated fusion of the PSVs with the plasma membrane and the subsequent secretion of their cargo, we purified this organelle from prespore cells. Analysis of protein extracts of highly purified PSVs indicated that, in addition to the cargo of structural spore coat proteins, many more proteins are associated with the PSVs. Their identification is paramount to the understanding of the mechanism of regulated secretion in this system. In this study we have taken the first comprehensive proteomic approach to the analysis of an entire, previously uncharacterized, organelle, with the goal of identifying the major proteins associated with the PSVs. We show that in addition to the structural spore coat proteins, the PSVs contain the enzymes needed for proper spore coat assembly (thioredoxin 2 and 3), regulatory proteins which we predict receive and transduce the developmental signal for secretion (rab7 GTPase, PI-3 kinase, NDP kinase and the calcium binding proteins calfumirin-1 and calreticulin) as well as proteins that interact with the cytoskeleton to mediate movement of the PSVs to the plasma membrane (actin binding proteins coactosin and profilin 1). In addition, the results suggest that proteins can play multiple roles in the cell, and that protein function can be dictated in part by subcellular localization. The identification of the PSV proteins is allowing us to develop testable hypotheses about the roles of these proteins within the functional context of developmentally regulated secretion.


N-Alkyl barbiturates. A series of compounds for the study of metabolic structure-activity relationships.
Many therapeutic agents are metabolised along multiple pathways, but up to now there have been few investigations addressing the question of which chemical features of drugs govern the participation in, and quantitative significance of, different biotransformation pathways. To assess the influence of variations of the chemical structure upon metabolism, a series of novel barbiturate analogues has been synthesized. The N1-monoalkylated and N1,N2-dialkylated phenobarbitones and 2-desoxyphenobarbitones have been synthesized via condensation of ethylphenylmalonic acid derivatives with different N-alkylated ureas or thioureas, and/or by base-catalyzed N-alkylation of different barbituric acids.