Structural Dynamics of Human Argonaute2 and Its Interaction with siRNAs Designed to Target Mutant tdp43.
The human Argonaute2 protein (Ago2) is a key player in RNA interference pathway and small RNA recognition by Ago2 is the crucial step in siRNA mediated gene silencing mechanism. The present study highlights the structural and functional dynamics of human Ago2 and the interaction mechanism of Ago2 with a set of seven siRNAs for the first time. The human Ago2 protein adopts two conformations such as "open" and "close" during the simulation of 25 ns. One of the domains named as PAZ, which is responsible for anchoring the 3'-end of siRNA guide strand, is observed as a highly flexible region. The interaction between Ago2 and siRNA, analyzed using a set of siRNAs (targeting at positions 128, 251, 341, 383, 537, 1113, and 1115 of mRNA) designed to target tdp43 mutants causing Amyotrophic Lateral Sclerosis (ALS) disease, revealed the stable and strong recognition of siRNA by the Ago2 protein during dynamics. Among the studied siRNAs, the siRNA341 is identified as a potent siRNA to recognize Ago2 and hence could be used further as a possible siRNA candidate to target the mutant tdp43 protein for the treatment of ALS patients.


Plasma levels of soluble receptor for advanced glycation end products and coronary atherosclerosis: possible correlation with clinical presentation.
Receptor for Advanced Glycation End-products (RAGE) is a multi-ligand receptor ubiquitous present on epithelial, neuronal, vascular and inflammatory cells, usually expressed at low levels in homeostasis and to increased degrees at sites of stress or injury. The aim of the present study was to evaluate sRAGE plasma levels in patients with Acute Coronary Syndrome (ACS) and to assess its diagnostic efficacy in identification of patients with acute events. Plasma levels of sRAGE were determined in 860 patients with Coronary Artery Disease (CAD): 530 patients presented stable angina and 330 were observed during acute ischemic event (147 with unstable angina and 183 with myocardial infarction). sRAGE plasma levels were significantly lower in patients with ACS than in patients with stable angina: [median 584 pg/mL (IQR: 266-851 pg/mL) in MI patients, median 769 pg/mL (IQR: 394-987 pg/mL) in patients with unstable angina, median 834 pg/mL (IQR 630-1005 pg/mL) in patients with stable angina; P < 0.001]. sRAGE levels did not differ among ACS patients stratified by the extent of coronary artery disease. In conclusion, this study confirm the role of sRAGE in activation and progression of inflammatory process and suggests the possibility that sRAGE can be considered an indicator of destabilization of vulnerable plaque.


Generation of double femtosecond pulses by using two transmissive gratings.
Double femtosecond laser pulses are usually generated by using a Michelson structure. We propose a novel and simple device by using two high-density transmissive gratings for generation of double pulses and conversion between single and double laser pulses by shifting one of the two gratings by a quarter period. The apparatus has the advantages of compact volume, simple structure, and convenience in manipulation. Experimental outputs of the double laser pulses are well verified in experiment, which can be properly explained by numerical simulation with the rigorous coupled-wave theory. This structure provides an interesting approach for generation of double pulses and conversion between single and double laser pulses for practical applications of the femtosecond laser.


[Detection of cardiac embolism by transcranial Doppler].
Cerebral emboli, regardless of their nature, can be detected by transcranial Doppler by performing long-term recordings of the middle cerebral artery. The signals detected, called HITS (high intensity transient signals) described in vitro and in vivo have particular physical characteristics. These embolic signals have been identified in patients with prosthetic heart valves, in patients with atrial fibrillation, following myocardial infarction, following cerebrovascular accident, in patients with carotid stenosis or aortic plaques, during cardiac and carotid surgery and during carotid angiography. During assessment of a cerebrovascular accident, the discovery of HITS could be an argument in favour of an embolic origin. This new technique raises great hopes for establishing the relationship between the discovery of emboligenic heart disease and their neurological clinical manifestations. This new technique has many fields of application: investigation of asymptomatic emboli in patients with emboligenic heart disease, efficacy of antiaggregant, anticoagulant or surgical treatments.


Synergistic Effect of Sr-O Divacancy and Exposing Facets in SrTiO3 Micro/Nano Particle: Accelerating Exciton Formation and Splitting, Highly Efficient Co2+ Photooxidation.
As a typical perovskite-type crystal, polyhedral strontium titanate (SrTiO3 ) has shown anisotropic charge transport behavior in recent studies, however, the carrier transportation and transition of which has not been explained very clearly. This work present the existence of Sr and O divacancies in the novel rhombicuboctahedron SrTiO3 micro/nano particles (Sr1- x TiO3- x /TiO2- x ) with exposing (100), (110) and (111) facets and the diameter of 300-700 nm synthesized via hydrothermal synthesis, and also summarizes the dissociation mechanism of self-trapped excitons (STEs) caused by the divacancy and facet effect. In addition, most importantly, the metastable STEs with ultra-low binding energy (Eb < 3 meV) under illumination are discovered. Combining the model of S-scheme heterojunction, a conversion mechanism of photoinduced carriers is proposed. The photocatalytic reaction of Co2+ is used as the probe reaction, and the unique Sr1- x TiO3- x /TiO2- x possesses a high photooxidation efficiency of Co2+ , by which 70.3% of Co2+ is oxidized to Co3+ (CoOOH) in 5 min. This finding may provide a guideline for an optimal design of the photocatalytic materials for the recovery and extraction of metal ions based on SrTiO3 .


[The sarcopenia: a multicomponent strategy for therapy (review of literature).]
Sarcopenia is an age-related loss of muscle mass, accompanied by a decrease in muscle function and/or strength, is increasingly recognized as a major risk factor for adverse outcomes in the weak elderly. The proven Association between hypodynamia, low-calorie diet, insufficient protein intake, and sarcopenia in older individuals suggests that exercise and targeted dietary supplements may have a significant therapeutic effect on the treatment and prevention of this condition. In this literature review, we summarize available data on the positive effects of various therapeutic interventions on muscle tissue.


In vitro analysis of antiangiogenic activity of fungi isolated from clinical cases of equine keratomycosis.
OBJECTIVE: The goal of this project was to explore the possibility that fungal organisms produce metabolites that inhibit angiogenesis. Procedures Fungal cultures were obtained from cases of keratomycosis, grown in Sabouraud's dextrose broth, and sterile filtered for use in experiments. The Matrigel assay was used to screen the filtrate samples for antiangiogenic activity. Matrigel is a basement membrane matrix that supports the differentiation of human umbilical vein endothelial (HUVE) cells into a capillary-like network of tubules. HUVE cells were cultured using standard techniques and passaged at confluence, with all cells being used at passage 3-6. HUVE cells (40 000 cells) were pipetted into each well of a 24-well tissue-culture plate coated with Matrigel. An aliquot of fungal media filtrate was added to each well and the plates allowed to incubate for 18 h, at which time they were evaluated for tubule formation. RESULTS: Two fungal isolates showed inhibition of tubule formation. The addition of 100, 200 and 400 &mgr;L of the fungal media filtrate from the first isolate (Fusarium sp. 99A34574) produced a consistent and dose-dependent inhibition of tubule formation. The second isolate (Aspergillus sp. 271599) did not show inhibition of tubule formation with 100 or 200 &mgr;L added to the wells, however, it did show inhibition at 400 &mgr;L/well. The remaining three isolates did not cause inhibition at any concentration. CONCLUSIONS: Our findings suggest that certain fungal organisms produce metabolites that inhibit tubule formation in vitro, and that these metabolites may play a significant role in altering the host vascular response to fungal infections of the cornea.


Perceived and Measured Physical Fitness of Police Students.
The physical fitness of police officers needs to support good health and physical performance. Physical fitness comprises a considerable amount of training for police students who are to become police officers. However, to what degree police students are able to perceive their fitness level and differentiate between health-related and performance-related physical fitness is unknown. Therefore, the first aim of this study was to investigate the possibility of differentiation between health-related and performance-related physical fitness using physical self-concept and measured indicators of physical fitness. The second aim of this study was to investigate the association between components of physical self-concept and measured indicators of physical fitness of police students. The sample of 177 police students of both sexes (98 males and 79 females) completed a 40-item physical self-description questionnaire and their physical abilities were assessed for handgrip strength, standing long jump, 30 s sit-ups, and 12-min running. Principal component analysis established health-related and performance-related physical fitness from both perceived and measured physical fitness measures. Correlation analysis revealed a significant relationship between the perceived and measured physical fitness. Results suggest small to moderate ability to recognize the level of certain physical abilities, indicating the association between psychological mechanisms and biological functioning.


Subcellular tracking of drug release from carbon nanotube vehicles in living cells.
The direct observation of drug release from carbon nanotube vehicles in living cells is realized through a unique two-dye labeling approach. Single-walled carbon nanotubes (SWNTs) are firstly marked with fluorescein isothiocyanate (FITC) to track their location and movement inside the cell. Then a fluorescent anticancer drug doxorubicin (DOX) is attached by means of π-stacking onto SWNTs. Delivered by SWNTs into cells, DOX will detach from the vehicle in an acidic environment due to the pH-dependent π-π stacking interaction between DOX and SWNTs. From observation of the two different kinds of fluorescence (green and red) that respectively represent the carrier SWNTs and drug DOX, the process of drug release inside the living cell can be monitored under a confocal microscope. Results show that the drug DOX detaches from SWNTs inside the lysosomes to yield free molecules and escape into the cytoplasm and finally into the nucleus, while the vehicle SWNTs are trapped inside the lysosomes, without entering the nucleus. The current observations confirm previously proposed mechanisms for drug/DOX release inside cells. The experimental establishment of drug-release mechanisms in living cells here might provide important insights for future design of new drug-delivery and release systems.


Rhesus macaques show increased resistance to repeated SHIV intrarectal exposure following a heterologous regimen of rVSV vector vaccine expressing HIV antigen.
Despite the human immunodeficiency virus (HIV) pandemic continuing worldwide for 40 years, no vaccine to combat the disease has been licenced for use in at risk populations. Here, we describe a novel recombinant vesicular stomatitis virus (rVSV) vector vaccine expressing modified HIV envelope glycoproteins and Ebola virus glycoprotein. Three heterologous immunizations successfully prevented infection by a different clade SHIV in 60% of non-human primates (NHPs). No trend was observed between resistance and antibody interactions. Resistance to infection was associated with high proportions of central memory T-cell CD69 and CD154 marker upregulation, increased IL-2 production, and a reduced IFN-γ response, offering insight into correlates of protection.


Activation of glycogen synthase kinase-3 beta is required for hyperdopamine and D2 receptor-mediated inhibition of synaptic NMDA receptor function in the rat prefrontal cortex.
The interactions between dopamine and glutamate systems play an essential role in normal brain functions and neuropsychiatric disorders. The mechanism of NMDA receptor regulation through high concentrations of dopamine, however, remains unclear. Here, we show the signaling pathways involved in hyperdopaminergic regulation of NMDA receptor functions in the prefrontal cortex by incubating cortical slices with high concentration of dopamine or administering dopamine reuptake inhibitor 1-(2-[bis-(4-fluorophenyl)methoxy]ethyl)- 4-(3-phenylpropyl)piperazine (GBR12909) in vivo. We found that, under both conditions, the synaptic NMDA receptor-mediated currents were significantly attenuated by excessive dopamine stimulation through activation of D(2) receptors. Furthermore, high dose of dopamine failed to affect NMDA receptor-mediated currents after blockade of NR2B subunits but triggered a dynamin-dependent endocytosis of NMDA receptors. The high-dose dopamine/D(2) receptor-mediated suppression of NMDA receptors was involved in the increase of glycogen synthase kinase-3beta (GSK-3beta) activity, which in turn phosphorylates beta-catenin and disrupts beta-catenin-NR2B interaction, but was dependent on neither Gq11 nor PLC (phospholipase C). Moreover, the hyperdopamine induced by GBR12909 significantly decreased the expression of both surface and intracellular NR2B proteins, as well as NR2B mRNA levels, suggesting an inhibition of protein synthesis. These effects were, however, completely reversed by administration of either GSK-3beta inhibitor or D(2) receptor antagonist. These results therefore suggest that GSK-3beta is required for the hyperdopamine/D(2) receptor-mediated inhibition of NMDA receptors in the prefrontal neurons and these actions may underlie D(2) receptor-mediated psychostimulant effects and hyperdopamine-dependent behaviors in the brain.


[Control of malaria in Africa: from the eradication of malaria to the control of malaria].
The control strategies usually recommended in Africa south of the Sahara have not been followed by a lasting decline in the endemic level of the disease in the rural environment. The set-backs of the eradication period have revealed the need to redefine control objectives and strategies adapted to each type of malaria. Systematic chemotherapy for attacks of fever is a strategy that can be proposed in all circumstances; it avoids mortality from malaria; it can be carried out in the field on a country scale. The other means of control, directed against the parasites and the vectors, may be used depending of the specific situation in each country and on each type of malaria. Malaria control has to be integrated into the basic health system of each country, and we need to improve health education and develop primary health care. We must continue with our efforts to develop new antimalarial drugs, insecticides and vaccines; urgent steps must be taken to train malariologists capable of adapting the new techniques to the actual situation in the field. It is by associating socioeconomic development with the control means as a whole that we shall be able to control the various types of malaria and to reach out towards the objective of health for all by the year 2000.


Use of midline catheters in low birth weight infants.
Prolonged intravenous access is an important consideration in the treatment of premature neonates. Because peripheral intravenous catheters have short dwell times, centrally positioned percutaneous catheters have been used increasingly in neonatal intensive care. Midline catheters are peripherally inserted long catheters advanced only to the proximal portion of a limb or to the neck, with the tip remaining outside the thoracic and abdominal cavities. Midline catheters were placed with use of an innovative technique via a 24-gauge Angiocath catheter (Becton Dickinson, Sandy, Utah). This study compared dwell time and reason for removal of midline catheters with respective data for peripheral intravenous catheters. Dwell time was found to be 9.0 +/- 1.4 days for the midline catheters versus 3.1 +/- 0.5 days for peripheral intravenous catheters placed in the same patients (p < 0.05). No significant complications occurred with the use of midline catheters. These data suggest that midline catheters may be useful in neonates who require a limited duration of intravenous therapy.


Assessing the therapeutic impact of a community group on a secure hospital ward.
The constructs of ward staff and patients in a medium secure rehabilitation ward were assessed before and after an eight-month trial of a twice weekly community group. One of the aims of this group was to address day-to-day interpersonal issues and to encourage patients to take social responsibility for their actions and to learn to support one another. Another aim was to encourage staff and patients to consider the possibility of applying some therapeutic community principles on the ward. These aims were assessed using repertory grid techniques. The results were that over the eight-month pilot both staff and patient groups shifted towards viewing their ideal ward as a therapeutic community (TC). However qualitative information suggested that considerable difficulties arose in bringing about the necessary cultural changes. As a result both staff and patients felt confused and disempowered. Future attempts are highly recommended, however better resourcing, particularly in the form of employing a trained full-time individual to co-ordinate activities, is essential.


Health-Related Quality of Life and Utility Scores of Lung Cancer Patients Treated with Traditional Chinese Medicine in China.
To assess health-related quality of life (HRQoL) and utility scores of lung cancer patients treated with traditional Chinese medicine (TCM) in China. This cross-sectional study included lung cancer patients treated with TCM in seven tertiary hospitals in Shanghai, China. The HRQoL and utility scores of these patients were measured using the five-level EQ-5D (EQ-5D-5L). The EQ-5D-5L utility scores were derived from the Chinese EQ-5D-5L Value Set. The relationships between HRQoL and the socio-demographic and clinical characteristics of these patients were further explored by Tobit regression. This study included a total of 347 patients. Their mean ± SD and median EQ-5D-5L utility scores were 0.851 ± 0.198 and 0.893, respectively. The highest proportion of participants reporting problems was observed in pain/discomfort dimension (57.9%) and anxiety/depression (45.5%). Lung cancer patients treated with TCM had poor HRQoL, influenced by cancer clinical stage. Lung cancer patients treated with TCM have poor HRQoL, with many patients reporting problems in the pain/discomfort and anxiety/depression dimensions. The information on health utility scores and HRQoL of lung cancer patients treated with TCM could be useful for future supportive care, economic evaluations and decision-making in China.


The effect of exposure to 35,000 ft on incidence of altitude decompression sickness.
Exposure to 35,000 ft without preoxygenation (breathing 100% oxygen prior to decompression) can result in severe decompression sickness (DCS). Exercise while decompressed increases the incidence and severity of symptoms. Clarification of the level of activity vs. time to symptom onset is needed to refine recommendations for current operations requiring 35,000-ft exposures. Currently, the U.S. Air Force limits these operations to 30 min following 75 min of preoxygenation. The objective of this study was to determine the effect of exercise intensity on DCS incidence and severity at 35,000 ft. Following 75 or 90 min of ground-level preoxygenation, 54 male and 38 female subjects were exposed to 35,000 ft for 3 h while performing strenuous exercise, mild exercise, or seated rest. The subjects were monitored for venous gas emboli (VGE) with an echo-imaging system and observed for signs and symptoms of DCS. Exposures involving strenuous and mild exercise resulted in higher incidence (p < 0.05) and earlier onset of symptoms (p < 0.05) of DCS than exposure at rest. Mild and strenuous exercise during exposure did not differ in incidence or rate of onset. Incidence at 30 min of exposure was 8% at rest and 23% while exercising. The results showed that current guidelines for 35,000-ft exposures keep DCS risk below 10% at rest. Exercise, even at mild levels, greatly increases the incidence and rate of onset of DCS.


Evaluation of a group-based behavioral intervention to promote adherence in adolescents with inflammatory bowel disease.
To pilot test the feasibility and acceptability of a family-based group behavioral intervention and to improve medication adherence in adolescents diagnosed with inflammatory bowel disease. Participants were 40 adolescents aged 11-18 years diagnosed with inflammatory bowel disease and their primary caregivers, who were randomized to either a four-session Family-Based Group Behavioral Treatment or Usual Care over a 6-week period. Adherence was measured using a multi-method, multi-informant assessment involving caregiver-report and patient-report, pill count data, and electronic monitoring. Adherence rates ranged from 66 to 89% for 6-mercaptopurine/azathioprine and 51 to 93% for mesalamine across assessment methods. The intervention was feasible, as evidenced by the 99% treatment session attendance rate, and acceptable based on patient and caregiver report. Repeated measures analysis of variance tests revealed nonsignificant differences between the conditions from baseline to post-treatment assessments for pill count, electronic monitor, and primary caregiver-reported adherence (P's>0.05). There was a statistically significant improvement in patient-reported mesalamine adherence represented by a significant main effect for Condition (F=22.24, P<0.01; δ=0.79) and Condition×Time interaction (F=13.32, P<0.05; δ=0.69). Findings suggest potential for use of behavioral intervention to improve medication adherence in this population. This intervention may be more effective with more complex regimens (e.g. multiple doses per day) such as those prescribed with mesalamine. Further research is needed to examine this type of intervention in more diverse samples with more active disease. Use of alternative adherence measurement approaches, including electronic pill boxes and/or real-time self-report (e.g. by text messaging, electronic diaries, etc.) is also recommended.


Isolation and molecular characterization of Mycobacterium bovis from Kafue lechwe (Kobus leche kafuensis) from Zambia.
Bovine tuberculosis (BTB) is a chronic bacterial disease caused by Mycobacterium bovis. Infections due to M. bovis, which serves as a stable reservoir, can pose serious challenge to control and eradicate in both wildlife and livestock at the interface. This study aimed at isolating and characterizing M. bovis from Kafue lechwe (Kobus leche kafuensis) and black lechwe (Kobus leche smithemani) at the animal/human interface in Zambia. The samples with lesions compatible with BTB collected during the hunting seasons of 2009 and 2010 were cultured for isolation of mycobacteria using Stonebrink with pyruvate (BD Diagnostics, MD, USA) and Middlebrook 7H10 (BD Diagnostics) slants. Isolated mycobacteria were identified using IS6110 polymerase chain reaction and deletion analysis. Molecular characterization of the isolates was performed using spoligotyping and mycobacteria interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) with nine loci. Data was analyzed using BioNumerics software 6.1. Out of the 39 samples, acid fast bacilli were detected in 27 (69.2 %) based on smear microscopy. Seven isolates were found to belong to Mycobacterium tuberculosis complex, and all were identified as M. bovis based on deletion analysis. All seven isolates were identical on spoligotyping as belonging to the SB0120 (SIT 482). MIRU-VNTR differentiated the isolates into five different patterns. This study has confirmed that M. bovis circulates in the Kafue lechwe, and non-tuberculous mycobacteria were detected in the black lechwe in Zambia which represents a wildlife reservoir, with a potential to spillover to cattle and humans. Isolates of M. bovis from lechwe antelopes are much conserved as only one spoligotype was detected. The study has shown that three loci differentiated fairly well. This option is cheap and less laborious, and hence a better option in resource-strained country like Zambia. The study further showed that some of the loci recommended by the European Reference Laboratory are not suitable for typing M. bovis in Zambia.


Personal meaning and eating disorder treatment: comment on Warren et al.
Comments on an article by C. S. Warren et al. (see record 2013-21802-001). Clinicians who treat eating disorders are reported by Warren, Schafer, Crowley, and Olivardia (2013, pp. 553-564) to have significant levels of burnout, with significant observed relationships to important factors such as experience in the field and recent patient mortality. The levels of burnout among those who persist in treating patients with eating disorders, however, were lower than expected, and clinicians with a personal history of eating disorders reported finding higher levels of meaning in their work. This comment explores the possibility that many eating disorder clinicians find satisfaction and meaning in this work for reasons that may not be immediately obvious. Additionally, it considers the possibility that some of the personal meaning derived from clinicians who report themselves "recovered" may be due to residual, unrecognized, or unreported emotional struggle surrounding eating and exercise that lends additional personal significance to this work.


[Mutations of p53 gene exons 5 to 8 in human ovarian cancer and its clinical significance].
To detect p53 mutations in human ovarian cancer and to look for correlations between p53 mutation and clinicopathologic features. Mutations in exons 5-8 of p53 gene in 41 patients with primary ovarian cancer were screened by polymerase chain reaction-single strand conformation polymorphism analysis with non-isotopic silver staining method. Mutations of p53 were found in 17 cases (41.5%); p53 mutation was not correlated with the age, the FIGO stage and the histologic type of the patients. However, it was associated with grade and lymph node metastasis in a univariate analysis. Mutations of p53 occur commonly in human ovarian cancer. p53 mutation is an early event in ovarian carcinogenesis and may serve as a genetic marker for early diagnosis and prognosis.


Effectiveness of liposomal-N-acetylcysteine against LPS-induced lung injuries in rodents.
Acute lung injury (ALI) and its most severe form, the acute respiratory distress syndrome (ARDS) are frequent complications in critically ill patients and are responsible for significant morbidity and mortality. So far, experimental evidence supports the role of oxidants and oxidative injury in the pathogenesis of ALI/ARDS. In this study, the antioxidant effects of conventional N-acetylcysteine (NAC) and liposomally entrapped N-acetylcysteine (L-NAC) were evaluated in experimental animals challenged with lipopolysaccharide (LPS). Rats were pretreated with empty liposomes, NAC, or L-NAC (25mg/kg body weight, iv); 4h later were challenged with LPS (E. coli, LPS 0111:B4) and sacrificed 20h later. Challenge of saline (SAL)-pretreated animals with LPS resulted in lung injury as evidenced by increases in wet lung weight (edema), increases in lipid peroxidation (marker of oxidative stress), decreases of lung angiotensin-converting enzyme (ACE) (injury marker for pulmonary endothelial cells) and increases in the pro-inflammatory eicosanoids, thromboxane B(2) and leukotriene B(4). The LPS challenge also increased pulmonary myeloperoxidase activity and chloramine concentrations indicative of neutrophil infiltration and activation of the inflammatory response. Pretreatment of animals with L-NAC resulted in significant increases in the levels of non-protein thiols and NAC levels in lung homogenates (p<0.05) and bronchoalveolar lavage fluids (p<0.001), respectively. L-NAC was significantly (p<0.05) more effective than NAC or empty liposomes in attenuating the LPS-induced lung injuries as indicated by the aforementioned injury markers. Our results suggested that the delivery of NAC as a liposomal formulation improved its prophylactic effectiveness against LPS-induced lung injuries.


Effect of dietary fiber on endogenous nitrogen flows in lactating dairy cows.
The effect of dietary fiber on endogenous N secretion was studied using a 15N isotope dilution technique in four fistulated Holstein cows. Two isonitrogenous diets differing only in fiber (NDF and ADF) content were used in a crossover design. One diet (HF) contained 37.4% NDF, while the other (LF) contained 23.3%. A new model was developed to estimate endogenous N secretions and losses for the preintestinal, intestinal, and the total sections of the gastrointestinal tract. Three precursor pools: TCA-soluble fraction of plasma, intestinal mucosa, and milk were compared. Although endogenous losses estimated with the model were numerically different for each precursor pool selected (TCA-soluble fraction > mucosa > milk), treatment effects were similar. As intestinal mucosa is probably closest to the precursor pool, these data are discussed. Non-urea N endogenous secretions contributed 13% of the duodenal N flow but were not affected by the fiber content of the diet. The nonurea N endogenous flow at the duodenum was comprised of approximately equal inputs from endogenous N direct, and that incorporated into the microbial biomass. Total endogenous N flows at the duodenum exceeded, by nearly twofold, estimated inputs of urea-N to microbial biomass. Metabolic fecal output averaged 17% of fecal N and was not affected by level of dietary fiber, but net losses from secretions occurring in the small intestine were higher with the low fiber diet. Overall, endogenous N secretions represented 30% of total digestive tract protein synthesis.


Mechanism of the six-electron reduction of nitrite to ammonia by cytochrome c nitrite reductase.
Cytochrome c nitrite reductase catalyzes the six-electron reduction of nitrite to ammonia without the release of potential reaction intermediates, such as NO or hydroxylamine. On the basis of the crystallographic observation of reaction intermediates and of density functional calculations, we present a working hypothesis for the reaction mechanism of this multiheme enzyme which carries a novel lysine-coordinated heme group (Fe-Lys). It is proposed that nitrite reduction starts with a heterolytic cleavage of the N-O bond which is facilitated by a pronounced back-bonding interaction of nitrite coordinated through nitrogen to the reduced (Fe(II)) but not the oxidized (Fe(III)) active site iron. This step leads to the formation of an [FeNO](6) species and a water molecule and is further facilitated by a hydrogen bonding network that induces an electronic asymmetry in the nitrite molecule that weakens one N-O bond and strengthens the other. Subsequently, two rapid one-electron reductions lead to an [FeNO](8) form and, by protonation, to an Fe(II)-HNO adduct. Hereafter, hydroxylamine will be formed by a consecutive two-electron two-proton step which is dehydrated in the final two-electron reduction step to give ammonia and an additional water molecule. A single electron reduction of the active site closes the catalytic cycle.


A case of repeated accidental inhalation contamination of a male subject with 137Cs.
Two consecutive incidents of internal contamination occurred within a 13-mo period when a male worker (56 y, 70 kg) briefly entered and stayed (during a breakdown of control apparatus for air contamination) in a radioisotope storeroom with air contaminated by an explosion of old ampules containing 137Cs solution. Monitoring of the first contamination began on day 34 and that of the second one within 1 h after inhalation. According to the two-exponential model, long-term biological half-times of 92 and 93 d were obtained for the first and second contaminations, respectively. The short-term biological half-time for the second contamination was calculated as 3.0 d. The mean value of the fraction of 137Cs excreted in daily urine to that in total excreta was 0.88 for the first contamination and 0.89 for the second one.


Increased reverse energy flux area when focusing a linearly polarized annular beam with binary plates.
We suggest an annular field with linear polarization and a binary sectored phase for the formation of the inverse energy flux with an increased area of action. The possibility of the formation of a reverse energy flux over large areas in the focal domain at tight focusing of a linearly polarized field with the binary phase is shown theoretically and numerically. Moreover, the area increases with the number m. The proposed approach is simple, because linearly polarized radiation is the most common among laser sources and does not require additional devices for polarization transformations. The binary phase also provides ease of manufacture of phase plates with a high diffraction efficiency and high damage threshold. The results will be useful in optical manipulations and processing.


Dynamics of water storage in mature subalpine Picea abies: temporal and spatial patterns of change in stem radius.
Internal water reserves in bark and foliage of trees contribute to transpiration (T) and play an essential role in optimizing water transport by buffering extreme peaks of water consumption. We examined patterns of stem shrinkage and their relationship to tree water dynamics. We measured fluctuations in root radius and stem radius at different stem heights, T of twigs at the top of the crown and sap flow velocities in stem sections of mature subalpine Norway spruce (Picea abies (L.) Karst.) trees over 2 years. The output of each sensor was coupled by physical functions to a mechanistic flow and storage model of tree water relations. The data verified the model-predicted lag in water storage depletion in response to the onset of transpiration and the lag increased with increasing distance from the crown periphery. Between the crown and stem base, the delay ranged from a few minutes to several hours, depending on microclimatic conditions and tree water status. Stem volume changes were proportional to the amount of water exchanged between the elastic tissues of the bark and the rigid xylem, indicating that the "peristaltic" wave of stem contraction along the flow path represented depletion of water stored in bark. On a daily basis, stems lost between 0.2 and 0.5% of their volume as a result of bark dehydration, corresponding to about 2 to 5 l of water. This water contributed directly to T. According to the model based on hydraulic principles, there are three main components underlying the dynamics of water storage depletion: flow resistance, storage capacities of needles and bark, and T of each tree section. The resistances and capacities were proportional to the response delay, whereas T in the lower parts of the tree was inversely proportional. The pattern of T within the crown depended on water intercepted by the branches. Because of these weather-dependent factors, there was no time constant for the response delay along the flow path. Nevertheless, the upper crown and the root section tended to have longer response delays per meter of flow path than the stem. The diurnal course of stem radius fluctuations represents the sum of all external and internal conditions affecting tree water relations; stem radius fluctuations, therefore, provide a sensitive measure of tree water status.


Laparoscopic distal pancreatectomy: the Brisbane experience of forty-six cases.
Laparoscopic distal pancreatectomy (LDP) is a safe alternative to conventional open distal pancreatectomy, with advantages that include smaller incisions, less pain, and shorter postoperative recovery. Despite these apparent advantages, however, uptake of the procedure has been slow, with only a handful of series published. All LDPs performed in Brisbane, Australia, over a 10-year period (May 1996 to June 2006) were retrospectively reviewed. Forty-six consecutive LDPs were performed. A variety of lesions were resected, including nine cancers. Twelve patients were converted for oncological (6) or technical reasons (6). The spleen was retained in 14/29 patients, either by main splenic vessel preservation (9) or solely supported by the short gastric vessels (5), resulting in inferior pole infarction in 2 patients. Overall morbidity was 39%, including 15% pancreatic fistula. All fistulas resolved after a median of 6 weeks without re-operation. A non-significant trend toward fewer fistulas with stapled rather than sutured stump closure was observed (13% vs 19%; p=0.43). Median operative duration and hospital stay were 157 min and 7 days, respectively. There was no mortality. LDP is a safe alternative to conventional resection for a wide range of lesions. As with open resection, pancreatic fistula is the dominant morbidity, but is generally indolent. While spleen preservation is often possible, care must be taken to avoid infarction of the inferior pole if the Warshaw technique is utilized.


The Effects of Streptomycin, Desiccation, and UV Radiation on Ice Nucleation by Pseudomonas viridiflava.
Streptomycin (100 micrograms per milliliter), desiccation (over CaSO(4)), and ultraviolet radiation (4500 microwatts per square centimeter at 254 nanometers for 15 minutes) reduced ice nucleation activity by Pseudomonas viridiflava strain W-1 as determined by freezing drops of the bacterial suspensions. Highest residual ice nucleation activity by dead cells was obtained by desiccation, although no freezing above -3.5 degrees C was detected. The rate and extent of loss of ice nucleation activity following streptomycin and ultraviolet treatment was affected by preconditioning temperature. At 21 degrees C and above, loss of activity by dead cells was rapid and irreversible.


Does the benzodiazepine antagonist Ro 15-1788 reverse the actions of picrotoxin and pentylenetetrazole on social and exploratory behaviour?
The effects of the benzodiazepine receptor antagonist, Ro 15-1788, were examined in combination with those of picrotoxin and pentylenetetrazole on social and exploratory behaviour in the rat. Both Ro 15-1788 (10 mg/kg) and picrotoxin (2 mg/kg) reduced social interaction, but there was no additive effect of these drugs together. Ro 15-1788 (10 and 20 mg/kg, which had no intrinsic effects in the holeboard alone) antagonized the reductions in exploratory head-dipping and motor activity produced by low (2 mg/kg), but not by high (4 mg/kg), doses of picrotoxin. In contrast, pentylenetetrazole (20 mg/kg) enhanced the reduction in social interaction produced by Ro 15-1788 (10 mg/kg), in accordance with previous findings that the reductions in spontaneous behaviour in the holeboard produced by pentylenetetrazole are enhanced by Ro 15-1788. These results show that there are clear functional interactions between these two compounds at the GABA-benzodiazepine receptor complex in the CNS, and that the nature of these interactions differs for picrotoxin and pentylenetetrazole.


User Identity Protection in EEG-Based Brain-Computer Interfaces.
A brain-computer interface (BCI) establishes a direct communication pathway between the brain and an external device. Electroencephalogram (EEG) is the most popular input signal in BCIs, due to its convenience and low cost. Most research on EEG-based BCIs focuses on the accurate decoding of EEG signals; however, EEG signals also contain rich private information, e.g., user identity, emotion, and so on, which should be protected. This paper first exposes a serious privacy problem in EEG-based BCIs, i.e., the user identity in EEG data can be easily learned so that different sessions of EEG data from the same user can be associated together to more reliably mine private information. To address this issue, we further propose two approaches to convert the original EEG data into identity-unlearnable EEG data, i.e., removing the user identity information while maintaining the good performance on the primary BCI task. Experiments on seven EEG datasets from five different BCI paradigms showed that on average the generated identity-unlearnable EEG data can reduce the user identification accuracy from 70.01% to at most 21.36%, greatly facilitating user privacy protection in EEG-based BCIs.


Transforming growth factor-beta 1 induces alpha-smooth muscle-actin expression in cultured human and monkey trabecular meshwork.
Transforming growth factor-beta 1 (TGF-beta 1) induces alpha-smooth muscle (sm)-actin expression and a contractile myofibroblast-like phenotype in a considerable number of different cell types. Since alpha-sm-actin is expressed in some of the resident human trabecular meshwork (TM) cells in situ, and TGF-beta 1 is synthesized by TM in vitro, alpha-sm-actin expression in TM might also be under the influence of TGF-beta 1. To assess this question, TM cultures were initiated from the eyes of three human donors and three cynomolgus monkeys. Various doses of TGF-beta 1 (0.5-5 ng ml-1) were added to confluent cultures from third to fourth passages. Experiments were performed in the presence of fetal calf serum or under chemically defined serum-free conditions. Four days after treatment, cells were stained immunocytochemically for alpha-sm-actin, and the number of positively labelled cells was quantitatively evaluated. In addition, reverse transcription polymerase chain reaction (PCR) was performed using oligonucleotide primers specific for alpha-sm-actin. In control cultures supplemented with serum, 19.0 +/- 9.4% of human meshwork cells, and 10.2 +/- 4.5% of monkey meshwork cells expressed immunoreactivity for alpha-sm-actin. In human cultures, this number was significantly higher in serum-free conditions (34.1 +/- 7.7%). Treatment with TGF-beta 1 induced alpha-sm-actin expression in a dose-dependent manner. At 5 ng ml-1 TGF-beta 1, 75.5 +/- 7.1% of human meshwork cells expressed distinct stress fibers that stained positively for alpha-sm-actin (P < or = 0.01). A similar albeit smaller increase in alpha-sm-actin positive cells was observed in monkey cultures following treatment with TGF-beta 1. These effects were seen with and without serum, but not when TGF-beta 1 was supplemented in the presence of neutralizing antibodies. In PCR experiments, a distinct product was amplified with cDNA derived from cells treated with 0.1 ng and 1 ng ml-1 TGF-beta 1, but not in control cultures. We conclude that TGF-beta 1 may play a role in differentiating TM cells towards a myofibroblast-like cell type by modulating the expression of alpha-sm-actin.


Lumbar spine intrathecal transplantation of neural precursor cells promotes oligodendrocyte proliferation in hot spots of chronic demyelination.
Experimental autoimmune encephalomyelitis (EAE) is a basic and reliable model used to study clinical and pathological hallmarks of multiple sclerosis (MS) in rodents. Several studies suggest neural precursor cells (NPCs) as a significant research tool while reporting that transplanted NPCs are a promising therapeutic approach to treating neurological disorders, such as MS. The main objective was to approach a preclinical, in vivo scenario of oligodendrogenesis with NPCs, targeting the main chronic demyelinated lumbosacral milieu of EAE, via the least invasive delivery method which is lumbar puncture. We utilized MOG35-55 peptide to induce EAE in C57BL/6 mice and prior to the acute relapse, we intervened with either the traceable GFP+ cellular therapy or saline solution in the intrathecal space of their lumbar spine. A BrdU injection, which enabled us to monitor endogenous proliferation, marked the endpoint 50 days post-induction (50 dpi). Neuropathology with high-throughput, triple immunofluorescent, and transmission electron microscopy (TEM) data were extracted and analyzed. The experimental treatment attenuated the chronic phase of EAE (50 dpi; score <1) following an acute, clinical relapse. Myelination and axonal integrity were rescued in the NPC-treated animals along with suppressed immune populations. The differentiation profile of the exogenous NPCs and endogenous BrdU+ cells was location-dependent where GFP+ -rich areas drove undifferentiated phenotypes toward the oligodendrocyte lineage. In situ oligodendrocyte enrichment was demonstrated through increased (p < 0.001) gap junction channels of Cx32 and Cx47, reliable markers for proliferative oligodendroglia syncytium. TEM morphometric analysis ultimately manifested an increased g-ratio in lumbosacral fibers of the recovered animals (p < 0.001). Herein, we suggest that a single, lumbar intrathecal administration of NPCs capacitated a viable cellular load and resulted in clinical and pathological amelioration, stimulating resident OPCs to overcome the remyelination failure in EAE demyelinating locale.


Effect of iron deficiency on Trichuris suis and Ascaris suum infections in pigs.
The objective of this experiment was to detect a possible interaction between iron deficiency and intestinal nematode infections. We report on a 2 x 2 study where thirty-one 10-week-old pigs fed a low or a normal iron diet were infected with both Trichuris suis (4500 eggs) and Ascaris suum (1200 eggs). No significant difference was detected between diet groups with respect to parasitological parameters for A. suum or the total number of adult T. suis recovered at necropsy 10 weeks p.i. However, in the low iron group T. suis were located more proximally and the worms were increased in length. A higher proportion of pigs with initial faecal egg excretion at 6 weeks p.i. was observed in the low iron group, indicating a shortened pre-patency period. Worm fecundity and total faecal egg excretion were also highest in the low iron group. A significant correlation was found between female worm length and fecundity. The peripheral eosinophil counts were diminished in the low iron host groups. The infected low iron group experienced more severe pathophysiological changes in terms of hypoalbuminaemia and decreases in erythrocyte volumes. A significant inverse correlation existed between iron content in the bone-marrow and liver (body) store. In conclusion, iron deficiency increased the severity of T. suis infection in pigs.


A Comparison Study on the Magneto-Responsive Properties and Swelling Behaviors of a Polyacrylamide-Based Hydrogel Incorporating with Magnetic Particles.
This work investigates the mechanical properties, microstructures, and water-swelling behavior of a novel hydrogel filled with magnetic particles. The nanoparticles of magnetite (Fe3O4) and the micro-particles of carbonyl iron (CI) were selected and filled into a polyacrylamide (PAAM) hydrogel matrix to create two types of magnetic hydrogels. The isotropy and anisotropy of magnetic hydrogels are also presented in this study. The isotropic samples were cured without applying a magnetic field (MF), and the anisotropic samples were cured by applying an MF in the direction perpendicular to the thickness of the samples. The effects of the size, content, and inner structures of magnetic particles on the magneto-responsive and swelling properties of magnetic hydrogels were investigated. It was found that the magnetorheological (MR) effect of anisotropic samples was apparently higher than that of isotropic samples, and the hydrogels with CI exhibited a noticeable MR effect than those with Fe3O4. The storage modulus can be enhanced by increasing the filler content and size, forming an anisotropic structure, and applying an external MF. In addition, the magnetic hydrogels also have a swelling ability that can be tuned by varying the content and size of the particle fillers.


Integrin α1 has a long helix, extending from the transmembrane region to the cytoplasmic tail in detergent micelles.
Integrin proteins are very important adhesion receptors that mediate cell-cell and cell-extracellular matrix interactions. They play essential roles in cell signaling and the regulation of cellular shape, motility, and the cell cycle. Here, the transmembrane and cytoplasmic (TMC) domains of integrin α1 and β1 were over-expressed and purified in detergent micelles. The structure and backbone relaxations of α1-TMC in LDAO micelles were determined and analyzed using solution NMR. A long helix, extending from the transmembrane region to the cytoplasmic tail, was observed in α1-TMC. Structural comparisons of α1-TMC with reported αIIb-TMC domains indicated different conformations in the transmembrane regions and cytoplasmic tails. An NMR titration experiment indicated weak interactions between α1-TMC and β1-TMC through several α1-TMC residues located at its N-terminal juxta-transmembrane region and C-terminal extended helix region.


Joint care: chasing the shadow from our most sun-filled days.
With 118,000 people dying of cancers of different kinds in England yearly both the patients and their families need support through this rite de passage period. Sadly, the majority are not receiving the care they need. Author reports on the UK's first district general hospital-based support team for the care of the dying.


Functional complementation of the yeast P-type H-ATPase, PMA1, by the Pneumocystis carinii P-type H-ATPase, PCA1.
The opportunistic fungus Pneumocystis is the etiologic agent of an interstitial plasma cell pneumonia that primarily afflicts immunocompromised individuals. Like other fungi Pneumocystis maintains a H(+) plasma membrane gradient to drive nutrient uptake and regulates intracellular pH by ATP-dependent proton efflux. Previously, we identified a Pneumocystis gene, PCA1, whose predicted protein product was homologous to fungal proton pumps. In this study, we show by functional complementation in a Saccharomyces strain whose endogenous PMA1 proton pump activity is repressed that the Pneumocystis PCA1 encodes a H(+)-ATPase. The properties of PCA1 characterized in this system closely resemble those of yeast PMA1. Yeast expressing PCA1 grow at low pH and are able to acidify the external media. Maximal enzyme activity (V(max)) and efficiency of substrate utilization (K(m)) in plasma membranes were nearly identical for PCA1 and PMA1. PCA1 contains an inhibitory COOH-terminal domain; removal of the final 40 amino acids significantly increased V(max) and growth at pH 6.5. PCA1 activity was inhibited by proton pump inhibitors omeprazole and lansoprazole, but was unaffected by H(+)/K(+)-ATPase inhibitor SCH28080. Thus, H(+) homeostasis in Pneumocystis is likely regulated as in other fungi. This work also establishes a system for screening PCA1 inhibitors to identify new anti-Pneumocystis agents.


Mixed-Effects Modeling Framework for Amsterdam and Copenhagen for Outdoor NO2 Concentrations Using Measurements Sampled with Google Street View Cars.
High-resolution air quality (AQ) maps based on street-by-street measurements have become possible through large-scale mobile measurement campaigns. Such campaigns have produced data-only maps and have been used to produce empirical models [i.e., land use regression (LUR) models]. Assuming that all road segments are measured, we developed a mixed model framework that predicts concentrations by an LUR model, while allowing road segments to deviate from the LUR prediction based on between-segment variation as a random effect. We used Google Street View cars, equipped with high-quality AQ instruments, and measured the concentration of NO2 on every street in Amsterdam (n = 46.664) and Copenhagen (n = 28.499) on average seven times over the course of 9 and 16 months, respectively. We compared the data-only mapping, LUR, and mixed model estimates with measurements from passive samplers (n = 82) and predictions from dispersion models in the same time window as mobile monitoring. In Amsterdam, mixed model estimates correlated rs (Spearman correlation) = 0.85 with external measurements, whereas the data-only approach and LUR model estimates correlated rs = 0.74 and 0.75, respectively. Mixed model estimates also correlated higher rs = 0.65 with the deterministic model predictions compared to the data-only (rs = 0.50) and LUR model (rs = 0.61). In Copenhagen, mixed model estimates correlated rs = 0.51 with external model predictions compared to rs = 0.45 and rs = 0.50 for data-only and LUR model, respectively. Correlation increased for 97 locations (rs = 0.65) with more detailed traffic information. This means that the mixed model approach is able to combine the strength of data-only mapping (to show hyperlocal variation) and LUR models by shrinking uncertain concentrations toward the model output.


[Immunohistochemical characterization of serotoninergic afferent pathways in the visual system of the rat].
Using the immunohistochemical PAP-method occurrence, distribution and terminal aborization of serotoninergic axons were investigated in the visual system of the albino rat. The present study reveals that the innervation varies in quantitative and qualitative manner in the different parts of the visual system. A sparsly serotoninergic innervation was observed in the dorsal lateral geniculate body (d lgb) and in the nucleus lateralis posterior. An intensive serotoninergic innervation was found in the lateral part of the ventral lateral geniculate body (v lgb) and in the area praetectalis. Serotoninergic axons show a lamina specific orientation in the colliculus superior and in the visual cortex. Our findings indicate that serotoninergic axons have an intensive terminal aborization in subcortical visual nuclei, which are more or less connected with oculomotoric functions.


Dispositional and situational coping and mental health among Palestinian political ex-prisoners.
We examined, first, differences in dispositional and situational coping, and psychological distress between political ex-prisoners and their matched controls, and second, coping effectiveness in protecting mental health from impacts of imprisonment and military trauma. Thirdly, we tested the hypothesis that compatibility ("goodness of fit") between dispositional and situational coping would predict low psychological distress. Participants were 184 men recruited from a Palestinian community sample, 92 were former political prisoners and 92 non-prisoners. The dispositional coping was assessed as a general response style to hypothetical stressors and situational coping as responses to their own traumatic experiences. Psychological distress was measured by SCL-90-R, and posttraumatic stress disorder, depression and somatoform symptoms by scales based on CIDI 2.1 diagnostic interview. The results showed that, compared to non-prisoners, the political ex-prisoners employed less avoidant, denying, and emotion-focused coping strategies. Military trauma was associated with avoidant and denying coping only among non-prisoners. The ex-prisoners showed more mental health and medical problems, especially when exposed to military trauma. None of the coping styles or strategies were effective in protecting the mental health in general or in either groups. However, main effect results revealed that the high level of active and constructive and low level of emotion-focused coping were associated with low levels of psychiatric symptoms and psychological distress.


Identification of (+) and (-) anti benzo[a]pyrene dihydrodiol epoxide-nucleic acid adducts by the 32P-postlabeling assay.
Purine deoxyribonucleoside 3'-phosphates were reacted with the (+)- and (-)-enantiomers of the anti dihydrodiol epoxide of benzo[a]pyrene. Products from cis and trans opening of the epoxide ring were separated by HPLC and they were identified by comparison of their CD spectra with those known for the corresponding nucleoside adducts. Thereafter, the eight known benzo[a]pyrene-purine deoxyribonucleoside-3'-phosphate adducts were postlabeled with [32P]ATP and T4 kinase and the positions of these individual bisphosphates were mapped by TLC. Though all eight adducts migrated to the same general region of the thin layer plates, the four possible adducts from each enantiomeric dihydrodiol epoxide were resolved.


Shionone Attenuates Sepsis-Induced Acute Kidney Injury by Regulating Macrophage Polarization via the ECM1/STAT5 Pathway.
To date, there are no specific drugs approved for the treatment of sepsis associated acute kidney injury (AKI). Shionone is a natural component with anti-inflammatory activity. In this study, we sought to determine the functional role of Shionone in sepsis-induced AKI. Animal models of AKI were constructed by cecum ligation and puncture (CLP) surgery. C57BL/6 mice were randomly assigned to the Sham, CLP, 10 mg/kg DXM, 50 mg/kg Shionone and 100 mg/kg Shionone groups. RAW264.7 treated with lipopolysaccharides (LPS) was used as an in vitro sepsis model and cells were divided into control, LPS, 1 μg/mL Shionone and 2 μg/mL Shionone groups. The pathological status was assessed by Hematoxylin-Eosin (HE) staining assay, protein expressions were detected by immunofluorescence staining and Western blot, macrophage typing was detected by flow, and the levels of pro-inflammatory factors (IL-6, IL-12, IL-1β, TNF-α) and anti-inflammatory factors (IL-10 and TGF-β) were measured using the corresponding kits. ECM1 is highly expressed in tissue-infiltrating macrophages under inflammatory conditions. It has been observed that Shionone inhibits the expression of ECM1 and attenuates sepsis-induced injury in kidney and inflammatory factor levels in serum. In addition, Shionone may reduce inflammatory factor levels through the promotion of M2 macrophages by GM-CSF/STAT5/Arg1 pathway to alleviate sepsis induced inflammation in vitro. These findings demonstrate that Shionone can alleviate sepsis-induced AKI by promoting M2 macrophage polarization through regulating the ECM1/STAT5 pathway.


The LOCalizer Radiofrequency Identification System: An Effective New Technology for Localizing Non-Palpable Breast Lesions for Surgery.
Background. Breast screening has decreased morbidity and mortality due to detection of early, non-palpable breast cancers. One of the challenges of performing breast-conserving surgery on non-palpable breast tumours is accurate localization of the cancer. We aimed to perform a feasibility study to examine the outcomes associated with the introduction of a novel radiofrequency identification system (RFID) called LOCalizer as an alternative to traditional wire-guided localization. Methods. Data were prospectively collected on all patients undergoing breast-conserving surgery using the LOCalizer RFID system in a regional cancer centre between July 2019 and March 2020. Patients had a RFID tag placed preoperatively and underwent surgical removal of the tag with the index lesion guided by a handheld LOCalizer probe. The primary aim was successful placement and retrieval of the RFID tag. Re-excision rates, specimen size, specimen weight, cancer subtype and complication rate were all recorded. Results. Sixty-nine patients aged between 50 and 69 years had a LOCalizer tag inserted between July 2019 and March 2020. Of these, 6 (8.7%) were diagnostic and 63 (91.3%) were therapeutic. There was no migration of RFID tags, and all tags were retrieved with the index lesion. The overall re-excision of margin rate was 17.4% (12/69). All re-excision of margins was due to positive radial margins. The overall complication rate was 1.4% with one grade 1 Clavien-Dindo morbidity. Conclusion. The LOCalizer RFID is an effective and safe wire-free localization method for non-palpable breast lesions.


Identification of a Novel Signature and Construction of a Nomogram Predicting Overall Survival in Clear Cell Renal Cell Carcinoma.
Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cell carcinoma (RCC), which accounts for majority of RCC-related deaths. It is clearly essential to further identify more novel prognostic signatures and therapeutic targets. We identified differentially expressed genes (DEGs) between ccRCC and adjacent normal tissues in GEO database using a Robust Rank Aggregation (RRA) method. An mRNA signature (mRNASig) based on DEGs was developed using Cox and LASSO analysis in the TCGA database and validated in the ICGC database. Afterward, the influence of mRNASig mRNAs on the immune microenvironment in ccRCC was explored using comprehensive bioinformatics analysis. A total of 957 robust DEGs were identified using the RRA method. mRNASig comprised CEP55, IFI44, NCF4, and TCIRG1 and was developed and validated to identify high-risk patients who had poorer prognosis than low-risk patients. A nomogram was also constructed based on mRNASig, AJCC stage, and tumor grade. The mRNASig were closely related to a variety of tumor-infiltrating lymphocytes, especially including CD8+ T cells, activated CD4+ memory T cells, regulatory T cells, activated NK cells, and resting NK cells. The mRNASig were also correlated positively with the expression of CTLA4, LAG3, PDCD1, TIGIT, and HAVCR2. We developed and validated mRNASig to assist clinicians in making personalized treatment decisions. Furthermore, CEP55, IFI44, NCF4, and TCIRG1 may be novel potential targets for future treatment of ccRCC.


Pinacidil relaxes porcine and human coronary arteries by activating ATP-dependent potassium channels in smooth muscle cells.
We investigated the effect of the potassium channel opener pinacidil on ATP-dependent K+ channels (KATP) in the relaxation of porcine and human coronary arteries by means of isometric contraction experiments in arterial rings. We also measured whole cell currents in freshly isolated porcine and human coronary artery vascular smooth muscle cells with patch clamp. We first characterized serotonin-induced precontractions in our vessels and proved that the contractions were mediated by Ca2+ influx through voltage-dependent Ca2+ channels. Similarly, we observed that serotonin-induced contractions were strongly enhanced by small K(+)-induced depolarizations. Pinacidil completely relaxed rings preconstricted with 5 microM serotonin and produced dose-dependent relaxations of 5 microM serotonin-preconstricted rings, with an IC50 of 1.26 microM. Similar results were observed (IC50 = 1.15 microM) when the endothelium was removed. The KATP blocker glibenclamide (3 microM), inhibited pinacidil-induced relaxations (5-10 microM) by approximately 25% although the KATP blocker tetrapentylammonium (10 microM), inhibited pinacidil-induced (5-10 microM) relaxations completely. Pinacidil 10 microM had only a minimal effect on rings precontracted with a 50 mM external K+ concentration (IC50 = 60 microM). Porcine and human arterial rings did not differ qualitatively in their responses. Moreover, in the patch clamp experiments pinacidil (1 microM and 20 microM) induced a large, nonrectifying, outward current in both human and porcine cells. The reversal potential was close to the K+ equilibrium potential, suggesting an induction of pinacidil-activated K+ current. The pinacidil-induced (1 microM) current was strongly inhibited by glibenclamide (3 microM). These data show that the relaxation of porcine and human coronary arteries by pinacidil is primarily induced by an opening of KATP in smooth muscle cells. Furthermore, the vasorelaxant effect of pinacidil is not endothelium dependent.


Manipulation of gene expression in the central nervous system with lentiviral vectors.
Viral vector-mediated gene transfer is widely used to manipulate gene expression (overexpression or knock down) in cultures and in different tissues of animals. Vectors based on lentiviruses have particularly useful features. Lentiviral vectors mediate gene transfer into any neuronal cell types and induce sustained expression without significant immune responses after delivery into the nervous system. Lentivirus-mediated expression of therapeutic genes has led to long-term treatment of animal models of neurological disorders, such as spinal injury, Parkinson's disease, Huntington's disease, and Alzheimer's disease. Here, we describe the preparation and purification of lentiviral vectors and methods of lentiviral infection in primary neural cultures and in brain regions of interest by stereotaxic injection.


β-Cell replacement as a treatment for type 1 diabetes: an overview of possible cell sources and current axes of research.
To efficiently treat type 1 diabetes, exogenous insulin injections currently represent the main approach to counter chronic hyperglycaemia. Unfortunately, such a therapeutic approach does not allow for perfectly maintained glucose homeostasis and, in time, cardiovascular complications may arise. Therefore, seeking alternative/improved treatments has become a major health concern as an increasing proportion of type 2 diabetes patients also require insulin supplementation. Towards this goal, numerous laboratories have focused their research on β-cell replacement therapies. Herein, we will review the current state of this research area and describe the cell sources that could potentially be used to replenish the depleted β-cell mass in diabetic patients.


Age-friendly neighbourhoods and physical activity of older Surinamese individuals in Rotterdam, the Netherlands.
Age-friendly neighbourhoods seem to promote physical activity among older individuals. Physical activity is especially important for chronically ill individuals. In the Netherlands, older Surinamese individuals are more likely to have chronic diseases than are their native Dutch counterparts. This study examined relationships of neighbourhood characteristics with physical activity among older Surinamese individuals in Rotterdam, the Netherlands. Of 2749 potential participants, 697 (25%) community-dwelling older (age ≥ 70 years) Surinamese individuals living in Rotterdam, the Netherlands, completed a questionnaire on personal and neighbourhood characteristics between March and June 2020. Correlation and multilevel regression analyses were performed to identify associations between missing neighbourhood characteristics for ageing in place and physical activity. Scores for the neighbourhood domains communication and information (r = -0.099, p ≤ 0.05), community support and health services (r = -0.139, p ≤ 0.001), and respect and social inclusion (r = -0.141, p ≤ 0.001), correlated negatively with participants' PA. In the multilevel analysis, overall missing neighbourhood characteristics to age in place scores were associated negatively with physical activity (p ≤ 0.05). This study showed the importance of age-friendly neighbourhoods for physical activity among older Surinamese individuals in Rotterdam, the Netherlands. Our findings suggest that the neighbourhood plays an important role in supporting older individuals' leading of physically active lifestyles. Further research is needed to support the development of interventions to create age-friendly neighbourhoods.


Recent advances in bovine vaccine technology.
A description of new commercial and experimental vaccines for viral and bacterial diseases of cattle can be broadly divided into those used for both beef and dairy cows and those used predominantly in dairy cattle. For both types of cattle, newer and experimental vaccines are directed against several of the important viral (e.g., bovine herpesvirus 1, bovine viral diarrhea virus, bovine respiratory syncytial virus, parainfluenza type 3, and foot-and-mouth disease virus) and bacterial pathogens (e.g., Pasteurella spp., Haemophilus somnus). The viral vaccines include gene-deleted, modified live, subunit, and peptide antigens. Newer bacterial vaccines, particularly those for Pasteurella spp., are composed of either modified-live vaccines or bacterins supplemented with toxoid or surface antigens. Haemophilus somnus vaccine research has concentrated mainly on defining unique surface antigens. Novel dairy cow vaccines would include the lipopolysaccharide-core (J5) antigen approach, which has been used for successful immunization against coliform mastitis. Core antigen vaccines also have reduced calf mortality from Gram-negative pathogens. Staphylococcal mastitis vaccines that contain capsular antigens, toxoids, or the staphylococcal fibronectin receptor are of active research interest. Vaccines against mastitis induced by Streptococcus agalactiae and Streptococcus uberis also are areas of intensive research. Delivery of multiple subunit antigens with optimal immune response induction has led to the investigation of attenuated heterologous viral and bacterial expression vectors such as bovine herpesvirus 1, vaccinia, and Salmonella spp. This discussion also demonstrates that molecular biology is being used to advance bovine vaccine technology.


Antibiofouling Polyvinylidene Fluoride Membrane Modified by Quaternary Ammonium Compound: Direct Contact-Killing versus Induced Indirect Contact-Killing.
Widespread applications of membrane technology call for the development of antibiofouling membranes. For the traditional contact-killing strategy, the antibacterial action is restricted to the surface: the membrane loses its antibiofouling efficacy once its surface is completely covered with a fouling layer. However, in this study, polyvinylidene fluoride (PVDF) microfiltration membranes blended with quaternary ammonium compound (QAC) exhibited a surprisingly lasting antimicrobial activity in the vicinity of the membrane surface. The results indicated that QAC was capable of driving surface segregation with a high structural stability, and the QAC modified membrane shows clear antibacterial effects against both Gram-positive and Gram-negative bacteria. Covering the modified membrane surface by an abiotic alginate layer resulted in a loss of antibacterial efficiency by 86.2%. In contrast, the antibacterial efficiency was maintained after developing a biofilm of Staphylococcus aureus of 30 μm in thickness. The current study may suggest that bacteria affected by contact-killing might interact with other bacteria in the vicinity, resulting in retarded biofilm growth. The antibiofouling effect and associated mechanism of the QAC modified membrane were further validated in a membrane bioreactor during long-term operation.


Entropy production in oscillatory processes during photosynthesis.
The flow of matter and heat and the rate of enzymatic reactions are examined using two models of photosynthesis that exhibit sustained and damped oscillatory dynamics, with the objective of calculating the rate of entropy generation and studying the effects of temperature and kinetic constants on the thermodynamic efficiency of photosynthesis. The global coefficient of heat transfer and the direct and inverse constants of the formation reaction of the RuBisCO-CO2 complex were used as control parameters. Results show that when the system moves from isothermal to non-isothermal conditions, the transition from a steady state to oscillations facilitates an increase in the energy efficiency of the process. The simulations were carried out for two photosynthetic models in a system on a chloroplast reactor scale.


Semaphorins as Potential Immune Therapeutic Targets for Cancer.
Semaphorins are a large class of secreted or membrane-bound molecules. It has been reported that semaphorins play important roles in regulating several hallmarks of cancer, including angiogenesis, metastasis, and immune evasion. Semaphorins and their receptors are widely expressed on tumor cells and immune cells. However, the biological role of semaphorins in tumor immune microenvironment is intricate. The dysregulation of semaphorins influences the recruitment and infiltration of immune cells, leading to abnormal anti-tumor effect. Although the underlying mechanisms of semaphorins on regulating tumor-infiltrating immune cell activation and functions are not fully understood, semaphorins can notably be promising immunotherapy targets for cancer.


Reproducibility of Baseline Tumour Metabolic Volume Measurements in Diffuse Large B-Cell LymphomA: Is There a Superior Method?
The metabolic tumour volume (MTV) is an independent prognostic indicator in diffuse large B-cell lymphoma (DLBCL). However, its measurement is not standardised and is subject to wide variations depending on the method used. This study aimed to compare the reproducibility of MTV measurement as well as the thresholds obtained for each method and their prognostic values. The baseline MTV was measured in 239 consecutive patients treated at Henri Becquerel Centre by two blinded evaluators. Eight methods were compared: 3 absolute (SUV (standardised uptake value) ≥ 2.5; SUV≥ liver SUVmax; SUV≥ PERCIST SUV), 1 percentage SUV threshold method (SUV ≥ 41% SUVmax) and 4 adaptive methods (Daisne, Nestle, Fitting, Black). The intraclass correlation coefficients were excellent, from 0.91 to 0.96, for the absolute SUV methods, Black and Nestle methods, and good for 41% SUVmax, Fitting and Daisne methods (0.82 to 0.88), with a significantly lower variability with absolute methods compared to 41% SUVmax (p < 0.04). Thresholds were found to be specific to each segmentation method and ranged from 295 to 552 cm3. There was a strong correlation between the MTV and patient prognosis regardless of the segmentation method used (p = 0.001 for PFS and OS). The largest inter-observer cut-off variability was observed in the 41% SUVmax method, which resulted in more inter-observer disagreements in the classification of patients between high and low MTV groups. MTV measurements based on absolute SUV criteria were found to be significantly more reproducible than those based on 41% SUVmax criteria. The threshold was specific for each of eight segmentation methods, but all predicted prognosis.


Outpatient Nonmelanoma Skin Cancer Excision and Reconstruction: A Clinical, Economical, and Patient Perception Analysis.
Nonmelanoma skin cancers constitute more than 15% of all types of cancer. To obtain the best cosmetic outcome, local flaps represent the ideal surgical choice. We conducted a retrospective review of patients treated from 2016 to 2019. The day-surgery group included 73 patients, and the outpatient group included 70 patients. We analyzed medical records regarding age of diagnosis, waiting time, site of lesion, reconstructive technique, histologic diagnosis, radicality of excision, and complications. We administered a survey based on quality items and carried out an economic evaluation. Outpatient removals were radical 92.6% of the time against the 78% of those performed in the operating room (P = 0.14). We observed two cases of wound dehiscence and two cases of hematoma in the day-surgery group. Economic analysis showed reduced costs in the outpatient setting. Skin excision and local flap reconstruction are safe procedures in an outpatient setting under a clinical, economical, and patient perception point of view.


Effects of diabetic keratopathy on corneal optical density, central corneal thickness, and corneal endothelial cell counts.
Diabetic keratopathy is an ocular complication that occurs with diabetes. In the present study, the effect of diabetic keratopathy on corneal optical density, central corneal thickness, and corneal endothelial cell count was investigated. One hundred and eighty diabetic patients (360 eyes) were enrolled in the study during the period from March, 2012 to March, 2013. The patients were divided into three age groups: <5, 5-10 and >10 years, with 60 patients per group (120 eyes). During the same period, 60 healthy cases (120 eyes) were selected and labeled as the normal control group. The Pentacam was used to measure the corneal optical density, and central corneal thickness. Specular microscopy was used to examine the corneal endothelial cell density. The coefficient of partial correlation was used to control age and correlate the analysis between the corneal optical density, corneal endothelial cell density, and central corneal thickness. The stage of the disease, the medial and intimal corneal optical density and central corneal thickness was analyzed in the diabetes group. The corneal optical density in the diabetes group increased compared with that of the normal control group. The medial and intimal corneal optical density and central corneal thickness were positively correlated with the course of the disease. However, the corneal endothelial cell density was not associated with the course of diabetes. There was a positive association between the medial and intimal corneal optical density and central corneal thickness of the diabetic patients. In conclusion, the results of the present study show that medial and intimal corneal optical density and central corneal thickness were sensitive indicators for early diabetic keratopathy.


Angioarchitecture predicting hemorrhage in cerebral arteriovenous malformations.
To find out lesions responsible for hemorrhage in arteriovenous malformations (AVMs), a retrospective study of angioarchitecture around the nidus was conducted in 27 patients who underwent conservative treatment. Comparison of angiograms revealed disappearance of an intranidal aneurysmal dilatation after the hemorrhagic events in two cases. The hematomas were adjacent to the dilatation, and no subarachnoid hemorrhage was evident. Obstruction of venous drainage, noticed in a case of spontaneous regression of AVM, was not demonstrated in the cases of hemorrhage. The intranidal aneurysmal dilatation is likely to have caused the hemorrhage in our cases. Careful endovascular embolization using proper materials should be indicated for an intranidal aneurysmal dilatation to prevent subsequent hemorrhage.


Diversity in the oligodendrocyte lineage: Plasticity or heterogeneity?
Heterogeneity is a widely recognized phenomenon within the majority of cell types in the body including cells of the central nervous system (CNS). The heterogeneity of neurons based on their distinct transmission modes and firing patterns has been recognized for decades, and is necessary to coordinate the immense variety of functions of the CNS. More recently, heterogeneity in glial cells has been identified, including heterogeneity in oligodendrocyte progenitor cells (OPCs) and oligodendrocytes. OPC subpopulations have been described based on their developmental origin, anatomical location in the grey or white matter, and expression of surface receptors. Oligodendrocytes are categorised according to differences in gene expression, myelinogenic potential, and axon specificity. Much of what is described as heterogeneity in oligodendrocyte lineage cells (OLCs) is based on phenotypic differences. However, without evidence for functional differences between putative subgroups of OLCs, distinguishing heterogeneity from plasticity and lineage state is difficult. Identifying functional differences between phenotypically distinct groups are therefore necessary for a deeper understanding of the role of OLCs in health and disease.


From Identification to Intelligence: An Assessment of the Suitability of Forensic DNA Phenotyping Service Providers for Use in Australian Law Enforcement Casework.
Forensic DNA Phenotyping (FDP) is an established but evolving field of DNA testing. It provides intelligence regarding the appearance (externally visible characteristics), biogeographical ancestry and age of an unknown donor and, although not necessarily a requirement for its casework application, has been previously used as a method of last resort in New South Wales (NSW) Police Force investigations. FDP can further assist law enforcement agencies by re-prioritising an existing pool of suspects or generating a new pool of suspects. In recent years, this capability has become ubiquitous with a wide range of service providers offering their expertise to law enforcement and the public. With the increase in the number of providers offering FDP and its potential to direct and target law enforcement resources, a thorough assessment of the applicability of these services was undertaken. Six service providers of FDP were assessed for suitability for NSW Police Force casework based on prediction accuracy, clarity of reporting, limitations of testing, cost and turnaround times. From these assessment criteria, a service provider for the prediction of biogeographical ancestry, hair and eye colour was deemed suitable for use in NSW Police Force casework. Importantly, the study highlighted the need for standardisation of terminology and reporting in this evolving field, and the requirement for interpretation by biologists with specialist expertise to translate the scientific data to intelligence for police investigators.


Lipid peroxidation and antioxidant status following thrombolytic therapy for acute myocardial infarction.
We have investigated the timescale of increased lipid peroxidation following successful early thrombolytic therapy for acute myocardial infarction and report for the first time reciprocal changes in plasma chain-breaking antioxidants. Sixty-seven patients were recruited following a first acute myocardial infarction within 6 h of the onset of symptoms and received 70 or 100 mg of recombinant tissue plasminogen activator (Actilyse) as two intravenous bolus injections 30 min apart. Serial blood samples were taken before administration of thrombolytic therapy and after 30 min, 60 min, 90 min, 6 h and 24 h. Coronary artery patency was assessed at 90 min by coronary angiography. Malondialdehyde (MDA), a marker of lipid peroxidation, and the chain-breaking antioxidants alpha-tocopherol, retinol and ascorbate were measured by high performance liquid chromatography. When the coronary artery was patent there was an early rise in plasma MDA (time 0 0.91 +/- 0.05 mumol.l-1) with levels peaking at 90 min (1.02 +/- 0.06, P < 0.05) and returning to baseline by 6 h (0.85 +/- 0.06), accompanied by reciprocal decreases in alpha-tocopherol (time 0 7.13 +/- 0.34 mumol.mmol-1 cholesterol, 90 min 6.64 +/- 0.33, P < 0.05) and retinol (time 0 1.99 +/- 0.10 mumol.l-1, 90 min 1.81 +/- 0.08, P < 0.05). Ascorbate levels did not change significantly until 24 h (time 0 29.5 +/- 4.9 mumol.l-1, 24 h 22.6 +/- 4.4, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Pathophysiology of microwave-induced traumatic brain injury.
Microwave technology has been widely used in numerous applications; however, excessive microwave exposure causes adverse effects, particularly in the brain. The present study aimed to evaluate the change in the number of neural cells and presence of apoptotic cells in rats for one month after exposure to excessive microwave radiation. The rats were exposed to 3.0 kW of microwaves for 0.1 sec and were sacrificed after 24 h (n=3), or 3 (n=3), 7 (n=3), 14 (n=3) or 28 days (n=4) of exposure. The neural cells were counted in the motor cortex and hippocampus [cornu ammonis 1 (CA1) and CA2] and the percentage of positive cells stained with terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) were also measured, which detected apoptotic cell death in the choroid plexus in the lateral ventricle, motor cortex and hippocampus. In the CA1, the number of neural cells decreased significantly by day 28 compared with that in the control (60.7 vs. 50.6, P=0.0358), but did not decrease before day 28. There were no significant differences on any day in the CA2 and the motor cortex. The number of cells showed a significant increase on day 7 compared to the control in the choroid plexus (2.1±1.1 vs. 21.8±19.1%, P=0.0318). There were no significant differences from the controls in the percentage of TUNEL-positive cells in the motor cortex and hippocampus. The effects of microwave exposure on the brain remain unclear; however, microwave-induced neurotrauma shows the same pathological changes as blast traumatic brain injury.


[Management of multiple pregnancies at the Befelatanana Antananarivo University Hospital Center (Madagascar): report of 143 cases].
Multiple pregnancies (MP) outcomes are often complicated. They deliver premature infants and provoke high blood pressure. A retrospective study was carried out in 1998 at the Maternity Hospital of Befelatanana, Antananarivo in order to assess MP frequency and to specify the most important favourising factors and difficulties during labor and the quality of the labor management. All pregnancies with a MP were included in this survey. 143 MP were registered: 142 twin pregnancies and 1 triplet pregnancy. 2.0 per cent of cases were recurrent MP. The average age of pregnancies was 26 years old. Among these 143 MP, 48.0 per cent were primiparas. Poor quality of prenatal visits is frequently encountered. As antecedents there are abortion, hormonal contraceptive taking, preterm delivery, gravidic toxemia, cicatricial uterus, ectopic pregnancy. 6.3 per cent of the first twin had breech presentation, 2.0 per cent transversal labor presentation. As events during labor 40.0 per cent dynamic dystocia, 26.0 per cent acute fetal suffering, 27.0 per cent hyperthermia, 23.0 per cent high blood pressure, some of them as eclampsia or pre-eclampsia were noted. 60.0 per cent of the first twin delivery were easy. Whatever his labor presentation, version by internal manipulations following by breech extraction was performed on the second twin (67.0 per cent of cases). 18.2 per cent of parturient women had cesaretomy. 11 maternal deaths were noted. Infant perinatal mortality rate was of 35.7 per cent. Infant morbidity and mortality are essentially due to infections. The authors conclude that complications prevention will be obtained by improvement of standard of living of all female able to procreate. It needs also correct cares at prenatal visits and during labor. Health education must be focalized on strict and correct surveillance of pregnancies and intergenesic periods by the reinforcement of planning family.


Herpes simplex virus type-2 glycoprotein-subunit vaccine: tolerance and humoral and cellular responses in humans.
Twenty-three subjects were administered three doses of inactivated herpes simplex virus (HSV) type 2 (HSV-2) glycoprotein-subunit vaccine, with doses 2 and 3 being given four and 22 weeks after dose 1. Both HSV-neutralizing antibody and antibody effective in antibody-dependent cell-mediated cytotoxicity were detected in serum samples of 12 of the 13 initially seronegative subjects by week 8. The geometric mean titers of both antibody types decreased between weeks 8 and 22 but increased one week after dose 3. HSV-neutralizing antibody remained detectable in serum samples of seven of the 10 initially seronegative subjects assayed seven months after dose 3. Cell-mediated immunity was detected by lymphocyte transformation responses to HSV-2 after vaccine administration in 12 of the 13 initially seronegative subjects. Unlike humoral antibody, which waned over time, in vitro cell-mediated immune responses remained stable over the seven-month follow-up period after dose 3. This HSV-2-subunit vaccine is well tolerated and elicits both humoral and cell-mediated immune responses to HSV.


Auxin biogenesis: subcellular compartmentation of indoleacetaldehyde reductases in cucumber seedlings.
Subcellular fractionation of cucumber (Cucumis sativus L.) seedlings was achieved, and two of the enzymes in the auxin biosynthetic pathway were localized. NADH-specific indoleacetaldehyde reductase activity was observed only in the cytosol fractions obtained from separated hypocotyl and cotyledon tissue. In contrast, a portion of the NADPH-specific indoleacetaldehyde reductase activity was associated with a microsomal fraction derived from these tissues. The NADPH-specific indoleacetaldehyde reductase was consistently found to be more firmly associated with the microsomal fraction derived from hypocotyls than with that from the cotyledons. These results indicate a division of the terminal steps of auxin biogenesis into at least two subcellular compartments.


Carbon Fixation Gradients across Spinach Leaves Do Not Follow Internal Light Gradients.
In situ measurements of 14C-CO2 incorporation into 40-[mu]m paradermal leaf sections of sun- and shade-grown spinach leaves were determined. Chlorophyll, carotenoid, and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) content in similar 40-[mu]m paradermal leaf sections was also measured. The carbon fixation gradient did not follow the leaf internal light gradient, which decreases exponentially across the leaf. Instead, the 14C-CO2 fixation was higher in the middle of the leaf. Contrary to expectations, the distribution of carbon fixation across the leaf showed that the spongy mesophyll contributes significantly to the total carbon reduced. Approximately 60% of the carboxylation occurred in the palisade mesophyll and 40% occurred in the spongy mesophyll. Carbon reduction correlated well with Rubisco content, and no correlation between chlorophyll and carotenoid content and Rubisco was observed in sun plants. The correlation among chlorophyll, carotenoids, Rubisco, and carbon fixation was higher in shade leaves than in sun leaves. The results are discussed in relation to leaf photosynthetic and biochemical measurements that generally consider the leaf as a single homogeneous unit.


An update on the function and regulation of methylerythritol phosphate and mevalonate pathways and their evolutionary dynamics.
Isoprenoids are among the largest and most chemically diverse classes of organic compounds in nature and are involved in the processes of photosynthesis, respiration, growth, development, and plant responses to stress. The basic building block units for isoprenoid synthesis-isopentenyl diphosphate and its isomer dimethylallyl diphosphate-are generated by the mevalonate (MVA) and methylerythritol phosphate (MEP) pathways. Here, we summarize recent advances on the roles of the MEP and MVA pathways in plant growth, development and stress responses, and attempt to define the underlying gene networks that orchestrate the MEP and MVA pathways in response to developmental or environmental cues. Through phylogenomic analysis, we also provide a new perspective on the evolution of the plant isoprenoid pathway. We conclude that the presence of the MVA pathway in plants may be associated with the transition from aquatic to subaerial and terrestrial environments, as lineages for its core components are absent in green algae. The emergence of the MVA pathway has acted as a key evolutionary event in plants that facilitated land colonization and subsequent embryo development, as well as adaptation to new and varied environments.


Association of childcare arrangement with overweight and obesity in preschool-aged children: a narrative review of literature.
The time children spend in childcare overlaps with daily meals and opportunities to be active. Thus these environments have the opportunity to promote-or hinder-healthy weight gain among children who attend them. The purpose of this narrative review was to compile findings from studies examining childcare type and weight outcomes among preschool-age children. A literature search was conducted using PubMed, PsychInfo and ERIC. Inclusion criteria were infant- to 5-year-old children exposed to any type of childcare with a cross-sectional or longitudinal weight outcome. Among 385 studies screened, 18 were included. For comparison across studies, type of childcare was categorized as: childcare center, Head Start, nanny/babysitter, non-relative care/family childcare home and relative care. Four studies found no association with childcare type and obesity, and 10 studies reported mixed results by type of care or subpopulation analyses. Two studies found an overall positive association, and two reported an inverse association. There were differences in direction of associations and findings by type of care arrangement. For Head Start, three of eight studies demonstrated a negative relationship with obesity; none demonstrated a positive association. No other childcare type demonstrated this inverse association. Informal types of care (relative and non-relative care in a home) were positively associated with child obesity in 3 of 10 studies. This association was less commonly reported among formal childcare centers (2 of 15 studies). The majority of studies, however, reported mixed findings or no association by childcare type. Results suggested no consistent evidence for a relationship between childcare and obesity risk, except Head Start. This review exposed the need for a consistent definition of childcare type and the exploration of unmeasured confounders, such as the nutrition and physical activity environment of childcare settings, to understand how they contribute to or protect against the development of overweight/obesity among children.


Absence of age effect on meiotic recombination between human X and Y chromosomes.
Recombination between the X and Y chromosomes is limited to the pseudoautosomal region and is necessary for proper segregation of the sex chromosomes during spermatogenesis. Failure of the sex chromosomes to disjoin properly during meiosis can result in individuals with a 47,XXY constitution, and approximately one-half of these result from paternal nondisjunction at meiosis I. Analysis of individuals with paternally derived 47,XXY has shown that the majority are the result of meiosis in which the X and Y chromosomes have failed to recombine. Our studies of sperm have demonstrated that aneuploid 24,XY sperm have a decreased recombination frequency, compared with that of normal sperm. Some studies have indicated a relationship of increased paternal age with 47,XXY offspring and with the production of XY disomic sperm, whereas others have failed to find such relationships. To determine whether there is a relationship between paternal age and recombination in the pseudoautosomal region, single-sperm genotyping was performed to measure the frequency of recombination between a sex-specific locus, STS/STS pseudogene, and a pseudoautosomal locus, DXYS15, in younger men (age < or =30 years) compared with older men (age > or =50 years). A total of 2,329 sperm cells were typed by single-sperm PCR in 20 men who were heterozygous for the DXYS15 locus (1,014 sperm from 10 younger men and 1,315 sperm from 10 older men). The mean recombination frequency was 39.2% in the younger men and 37.8% in the older men. There was no heterogeneity in the frequency of recombination rates. There was no significant difference between the recombination frequencies among the younger men and those among the older men, when analyzed by the clustered binomial Z test (Z=.69, P=.49). This result suggests that paternal age has no effect on the recombination frequency in the pseudoautosomal region.


Genetic evidence that two independent S-loci control RNase-based self-incompatibility in diploid strawberry.
The self-incompatibility mechanism that reduces inbreeding in many plants of the Rosaceae is attributed to a multi-allelic S locus which, in the Prunoideae and Maloideae subfamilies, comprises two complementary genes, a stylar-expressed S-RNase and a pollen-expressed SFB. To elucidate incompatibility in the subfamily Rosoideae, stylar-specific RNases and self-(in)compatibility status were analysed in various diploid strawberries, especially Fragaria nubicola and F. viridis, both self-incompatible, and F. vesca, self-compatible, and in various progenies derived from them. Unexpectedly, two unlinked RNase loci, S and T, were found, encoding peptides distinct from Prunoideae and Maloideae S-RNases; the presence of a single active allele at either is sufficient to confer self-incompatibility. By contrast, in diploid Maloideae and Prunoideae a single locus encodes S-RNases that share several conserved regions and two active alleles are required for self-incompatibility. Our evidence implicates the S locus in unilateral inter-specific incompatibility and shows that S and T RNases can, remarkably, confer not only allele-specific rejection of cognate pollen but also unspecific rejection of Sn Tn pollen, where n indicates a null allele, consistent with the the presence of the pollen component, SFB, activating the cognitive function of these RNases. Comparison of relevant linkage groups between Fragaria and Prunus suggests that Prunus S-RNases, unique in having two introns, may have resulted from gene conversion in an ancestor of Prunus. In addition, it is shown that there is a non-S locus that is essential for self-incompatibility in diploid Fragaria.


Complete nucleotide sequence of the penicillin G acylase gene and the flanking regions, and its expression in Escherichia coli.
The pga gene coding for penicillin G acylase (PGA) in Escherichia coli ATCC11105 was cloned, and its complete nucleotide sequence including 5'- and 3'-flanking regions was determined. Two nonidentical subunits that constitute an active PGA enzyme complex are known to be formed by processing of a common precursor molecule [Böck et al., FEMS Microbiol. Lett. 20 (1983) 141-144]. This novel type of protein processing was confirmed by a nucleotide sequencing study together with amino acid sequencing of two PGA subunits. In addition, it was found that the initiation codon, AUG, is preceded by an authentic ribosome-binding site, a consensus promoter sequence and putative cAMP receptor protein (CRP)-binding sites, and that the termination codon, UAA, is followed by a putative transcriptional terminator. The promoter function was confirmed by galactokinase assay using galK fusion plasmids. A recombinant plasmid was constructed to overproduce the enzyme using phage lambda pL promoter. Unexpectedly, thermal induction led to accumulation of the 94-kDa polypeptide rather than active PGA in large amounts. Western immunoblot analysis showed that this large polypeptide is the real precursor of PGA. It is evident, therefore, that the synthesis of active PGA in E. coli is affected by growth temperature and that the precursor processing step(s) is temperature-sensitive.