An entirely cell-based system to generate single-chain antibodies against cell surface receptors.
The generation of recombinant antibodies (Abs) using phage display is a proven method to obtain a large variety of Abs that bind with high affinity to a given antigen. Traditionally, the generation of single-chain Abs depends on the use of recombinant proteins in several stages of the procedure. This can be a problem, especially in the case of cell-surface receptors, because Abs generated and selected against recombinant proteins may not bind the same protein expressed on a cell surface in its native form and because the expression of some receptors as recombinant proteins is problematic. To overcome these difficulties, we developed a strategy to generate single-chain Abs that does not require the use of recombinant protein at any stage of the procedure. In this strategy, stably transfected cells are used for the immunization of mice, measuring Ab responses to immunization, panning the phage library, high-throughput screening of arrayed phage clones, and characterization of recombinant single-chain variable regions. This strategy was used to generate a panel of single-chain Abs specific for the innate immunity receptor Toll-like receptor 2. Once generated, individual single-chain variable regions were subcloned into an expression vector allowing the production of recombinant Abs in insect cells, thus avoiding the contamination of recombinant Abs with microbial products. This cell-based system efficiently generates Abs that bind to native molecules on the cell surface, bypasses the requirement of recombinant protein production, and avoids risks of microbial component contamination.


Making connections--strategies for single molecule fluorescence biophysics.
Fluorescence spectroscopy and fluorescence microscopy carried out on the single molecule level are elegant methods to decipher complex biological systems; it can provide a wealth of information that frequently is obscured in the averaging of ensemble measurements. Fluorescence can be used to localise a molecule, study its binding with interaction partners and ligands, or to follow conformational changes in large multicomponent systems. Efficient labelling of proteins and nucleic acids is very important for any fluorescence method, and equally the development of novel fluorophores has been crucial in making biomolecules amenable to single molecule fluorescence methods. In this paper we review novel coupling strategies that permit site-specific and efficient labelling of proteins. Furthermore, we will discuss progressive single molecule approaches that allow the detection of individual molecules and biomolecular complexes even directly isolated from cellular extracts at much higher and much lower concentrations than has been possible so far.


Accuracy and bias in the perception of the partner in a close relationship.
Partners in close relationships can be both accurate and biased in their perceptions of each other. Moreover, sometimes a bias can lead to accuracy. The authors describe a paradigm for the simultaneous measurement of accuracy and bias in 2-person relationships. One prevalent bias in close relationships is assumed similarity: Does the person think that his or her partner sees the world as he or she does? In a study of 238 dating and married heterosexual couples, the authors found evidence for both bias and accuracy: the bias effects were considerably stronger, especially when the measure was linked to the relationship. They found little or no evidence for gender differences in accuracy and bias.


Retention of spliceosomal components along ligated exons ensures efficient removal of multiple introns.
The majority of mammalian pre-mRNAs contains multiple introns that are excised prior to export and translation. After intron excision, ligated exon intermediates participate in subsequent intron excisions. However, exon ligation generates an exon of increased size, a feature of pre-mRNA splicing that can interfere with downstream splicing events. These considerations raise the question of whether unique mechanisms exist that permit efficient removal of introns neighboring ligated exons. Kinetic analyses of multiple intron-containing pre-mRNAs revealed that splicing is more efficient following an initial intron removal event, suggesting that either the recruitment of the exon junction complex (EJC) to ligated exons increases the efficiency of multiple intron excisions or that the initial definition of splice sites is sufficient to permit efficient splicing of introns neighboring ligated exons. Knockdown experiments show that the deposition of the EJC does not affect subsequent splicing kinetics. Instead, spliceosomal components that are not involved in the initial splicing event remain associated with the pre-mRNA to ensure efficient removal of neighboring introns. Thus, ligated exons do not require redefinition, providing an additional kinetic advantage for exon defined splice sites.


Perinatal periods of risk: analytic preparation and phase 1 analytic methods for investigating feto-infant mortality.
The Perinatal Periods of Risk (PPOR) methods provide the necessary framework and tools for large urban communities to investigate feto-infant mortality problems. Adapted from the Periods of Risk model developed by Dr. Brian McCarthy, the six-stage PPOR approach includes epidemiologic methods to be used in conjunction with community planning processes. Stage 2 of the PPOR approach has three major analytic parts: Analytic Preparation, which involves acquiring, preparing, and assessing vital records files; Phase 1 Analysis, which identifies local opportunity gaps; and Phase 2 Analyses, which investigate the opportunity gaps to determine likely causes of feto-infant mortality and to suggest appropriate actions. This article describes the first two analytic parts of PPOR, including methods, innovative aspects, rationale, limitations, and a community example. In Analytic Preparation, study files are acquired and prepared and data quality is assessed. In Phase 1 Analysis, feto-infant mortality is estimated for four distinct perinatal risk periods defined by both birthweight and age at death. These mutually exclusive risk periods are labeled Maternal Health and Prematurity, Maternal Care, Newborn Care, and Infant Health to suggest primary areas of prevention. Disparities within the study community are identified by comparing geographic areas, subpopulations, and time periods. Excess mortality numbers and rates are estimated by comparing the study population to an optimal reference population. This excess mortality is described as the opportunity gap because it indicates where communities have the potential to make improvement.


Disparity in cancer prevention and screening in aboriginal populations: recommendations for action.
Historically, cancer has occurred at a lower rate in aboriginal populations; however, it is now dramatically increasing. Unless preventive measures are taken, cancer rates among aboriginal peoples are expected to soon surpass those in non-aboriginal populations. Because a large proportion of malignant disorders are preventable, primary prevention through socioeconomic interventions, environmental changes, and lifestyle modification might provide the best option for reducing the increasing burden of cancers. Such efforts can be further amplified by making use of effective cancer screening programs for early detection of cancers at their most treatable stage. However, compared with non-aboriginal Canadians, many aboriginal Canadians lack equal access to cancer screening and prevention programs. In this paper, we discuss disparities in cancer prevention and screening in aboriginal populations in Canada. We begin with the relevant definitions and a theoretical perspective of disparity in health care in aboriginal populations. A framework of health determinants is proposed to explain the pathways associated with an increased risk of cancer that are potentially avoidable. Major challenges and knowledge gaps in relation to cancer care for aboriginal populations are addressed, and we make recommendations to eliminate disparities in cancer control and prevention.


Functional differences between human and yeast protein disulfide isomerase family proteins.
Previously, it has been reported that a mammalian protein disulfide isomerase (PDI), when expressed on a single copy number plasmid, can rescue growth of a PDI1-disrupted yeast. However, here, for the first time we demonstrated by tetrad analysis that human PDI (hPDI) is unable to replace yeast PDI (yPDI) when hPDI cDNA is integrated into the yeast chromosome. This observation indicates that hPDI is not functionally equivalent to yPDI. Estimation of the actual copy number of the plasmid, as well as comparison of isomerase and chaperone activities between human and yeast PDI homologues, indicates that one copy of hPDI cDNA is not sufficient to rescue the PDI1-disrupted strain. Notably, the isomerase activities of yPDI family proteins, Mpd1p, Mpd2p, and Eug1p, were extremely low, although yPDI itself exhibited twice as much isomerase activity as hPDI in vitro. Moreover, with the exception of Mpd1p, all hPDI and yPDI family proteins had chaperone activity, this being particularly strong in the case of yPDI and Mpd2p. These observations indicate that the growth of Saccharomyces cerevisiae is completely dependent on the isomerase activity of yPDI.


Molecular analysis of the hotspot of recombination in the murine major histocompatibility complex.
Biological and serological assays have been used to define four subregions for the I region of the major histocompatibility complex (MHC) in the order I-A, I-B, I-J, and I-E. The I-J subregion presumably encodes the I-J polypeptide of the elusive T-cell suppressor factors. Restriction enzyme site polymorphisms and DNA sequence analyses of the I region from four recombinant mouse strains were used to localize the putative I-B and I-J subregions to a 1.0-kilobase (kb) region within the E beta gene. Sequencing this region from E beta clones derived from the two mouse strains: B10.A(3R), I-Jb and B10.A(5R), I-Jk initially used to define the I-J subregion revealed that these regions are identical, hence the distinct I-Jb and I-Jk molecules cannot be encoded by this DNA. In addition, the DNA sequence data also refute the earlier mapping of the I-B subregion. Analysis of the DNA sequences of three parental and four I region recombinants reveals that the recombinant events in three of the recombinant strains occurred within a 1-kb region of DNA, supporting the proposition that a hotspot for recombination exists in the I region. The only striking feature of this hotspot is a tetramer repeat (AGGC)n that shows 80 percent homology to the minisatellite sequence which may facilitate recombination in human chromosomes.


A consensus guide to capturing the ability to inhibit actions and impulsive behaviors in the stop-signal task.
Response inhibition is essential for navigating everyday life. Its derailment is considered integral to numerous neurological and psychiatric disorders, and more generally, to a wide range of behavioral and health problems. Response-inhibition efficiency furthermore correlates with treatment outcome in some of these conditions. The stop-signal task is an essential tool to determine how quickly response inhibition is implemented. Despite its apparent simplicity, there are many features (ranging from task design to data analysis) that vary across studies in ways that can easily compromise the validity of the obtained results. Our goal is to facilitate a more accurate use of the stop-signal task. To this end, we provide 12 easy-to-implement consensus recommendations and point out the problems that can arise when they are not followed. Furthermore, we provide user-friendly open-source resources intended to inform statistical-power considerations, facilitate the correct implementation of the task, and assist in proper data analysis.


An automatic diagnostic system for CT liver image classification.
Computed tomography (CT) images have been widely used for liver disease diagnosis. Designing and developing computer-assisted image processing techniques to help doctors improve their diagnosis has received considerable interests over the past years. In this paper, a CT liver image diagnostic classification system is presented which will automatically find, extract the CT liver boundary and further classify liver diseases. The system comprises a detect-before-extract (DBE) system which automatically finds the liver boundary and a neural network liver classifier which uses specially designed feature descriptors to distinguish normal liver, two types of liver tumors, hepatoma and hemageoma. The DBE system applies the concept of the normalized fractional Brownian motion model to find an initial liver boundary and then uses a deformable contour model to precisely delineate the liver boundary. The neural network is included to classify liver tumors into hepatoma and hemageoma. It is implemented by a modified probabilistic neural network (PNN) [MPNN] in conjunction with feature descriptors which are generated by fractal feature information and the gray-level co-occurrence matrix. The proposed system was evaluated by 30 liver cases and shown to be efficient and very effective.


Correlates of existential well-being and their association with health-related quality of life in breast cancer survivors compared with the general population.
The aim was to evaluate the correlates of existential well-being (EWB) and investigate the relationship between EWB and health-related quality of life (HRQOL) according to the McGill Quality of Life Questionnaire (MQOL) in breast cancer survivors (BCS) and in the general population. BCS (N = 1,933) recruited from five large hospitals completed a mailed survey, which included the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (QLQ-C30), Quality of Life Questionnaire Breast Cancer Module (QLQ-BR23) and the MQOL. Reference data were derived from a representative sample of the Korean adult population composed of 500 women selected by a nationwide random route technique. Multiple logistic regression analysis showed that members of the general population with a lower educational status were more likely to report poor EWB. However, among BCS, correlates that were associated with poor EWB included lower monthly income, lower educational status, unemployment and comorbidities. Almost all factors listed on the HRQOL questionnaire were correlated with EWB in BCS as well as in the general population (P < 0.001). Factors listed on the QLQ-C30 that were correlated to a clinically meaningful extent with EWB in the general population included physical functioning, nausea and vomiting, and appetite loss, whereas among BCS, insomnia and distress over hair loss were meaningfully correlated with EWB. Compared with general population, BCS showed different EWB correlates and distinct associations with HRQOL findings. Our data suggest that socio-economic status and comorbidities influence on their EWB.


Altered ErbB4 splicing and cortical parvalbumin interneuron dysfunction in schizophrenia and mood disorders.
Working memory requires the activity of parvalbumin (PV) interneurons in the dorsolateral prefrontal cortex (DLPFC). Impaired working memory and lower PV expression in the DLPFC are reported in schizophrenia and to a lesser degree in mood disorders. We previously proposed that activity-dependent PV expression is lower in schizophrenia due to a shift in the splicing of erb-b2 receptor tyrosine kinase 4 (ErbB4) transcripts from major to inactive minor variants that reduces excitatory drive to PV interneurons. Here, we tested the hypothesis that the degree of major-to-minor shift in ErbB4 splicing predicts the level of PV expression across schizophrenia and mood disorders. Levels of ErbB4 splice variants and PV mRNA were quantified by PCR in the DLPFC from 40 matched tetrads (N = 160 subjects) of schizophrenia, bipolar disorder (BD), major depressive disorder (MDD), and unaffected comparison subjects. Relative to unaffected comparison subjects, the magnitude of increases in minor variant levels and decreases in major variant levels was greatest in schizophrenia, intermediate in BD, and least in MDD. The same rank order was present for the magnitude of increases in the composite splicing score, which reflects the degree of major-to-minor shift across all ErbB4 splice loci, and for the magnitude of deficient PV expression. Finally, the composite splicing score negatively predicted PV expression across all subject groups. Together, these findings demonstrate a shared relationship between ErbB4 splicing and PV expression and suggest that scaling of the major-to-minor shift in ErbB4 splicing may influence the severity of deficient PV interneuron activity across diagnoses.


Overexpression of SENP3 in oral squamous cell carcinoma and its association with differentiation.
Small ubiquitin-like modifier (SUMO) modification is an important post-translational protein modification that can be reversed by SUMO-specific proteases (SENPs); however, the physiological function of SENPs remains largely unexplored, and little is known about the regulation of SENPs themselves. As one of the crucial members of the SUMO system, SENP3 is essential for rRNA processing and cell proliferation. In the present study, we analysed the expression of SENP3 in human oral squamous cell carcinoma (OSCC) and investigated the correlation between its expression and clinicopathological parameters in OSCC patients. The expression of SENP3 was higher in OSCC tissues than that in the normal mucosa adjacent to the tumor, and a modest increase in reactive oxygen species (ROS) regulated SENP3 stability and localization. ROS induced SENP3 redistribution from the nucleoli to the nucleoplasm. Taken together, these results indicated that the expression level of SENP3 may be associated with the differentiation of OSCC and that SENP3 may play an important role in the development of OSCC under oxidative stress.


Understanding the glycome: an interactive view of glycosylation from glycocompositions to glycoepitopes.
Nowadays, due to the advance of experimental techniques in glycomics, large collections of glycan profiles are regularly published. The rapid growth of available glycan data accentuates the lack of innovative tools for visualizing and exploring large amount of information. Scientists resort to using general-purpose spreadsheet applications to create ad hoc data visualization. Thus, results end up being encoded in publication images and text, while valuable curated data is stored in files as supplementary information. To tackle this problem, we have built an interactive pipeline composed with three tools: Glynsight, EpitopeXtractor and Glydin'. Glycan profile data can be imported in Glynsight, which generates a custom interactive glycan profile. Several profiles can be compared and glycan composition is integrated with structural data stored in databases. Glycan structures of interest can then be sent to EpitopeXtractor to perform a glycoepitope extraction. EpitopeXtractor results can be superimposed on the Glydin' glycoepitope network. The network visualization allows fast detection of clusters of glycoepitopes and discovery of potential new targets. Each of these tools is standalone or can be used in conjunction with the others, depending on the data and the specific interest of the user. All the tools composing this pipeline are part of the Glycomics@ExPASy initiative and are available at https://www.expasy.org/glycomics.


Oxidative stress stimulates autophagic flux during ischemia/reperfusion.
Autophagy is a bulk degradation process in which cytosolic proteins and organelles are degraded through lysosomes. To evaluate autophagic flux in cardiac myocytes, we generated adenovirus and cardiac-specific transgenic mice harboring tandem fluorescent mRFP-GFP-LC3. Starvation significantly increased the number of mRFP-GFP-LC3 dots representing both autophagosomes and autolysosomes per cell, suggesting that autophagic flux is increased in cardiac myocytes. H(2)O(2) significantly increased autophagic flux, which was attenuated in the presence of N-2-mercaptopropionyl glycine (MPG), an antioxidant, suggesting that oxidative stress stimulates autophagy in cardiac myocytes. Myocardial ischemia/reperfusion (I/R) increased both autophagosomes and autolysosomes, thereby increasing autophagic flux. Treatment with MPG attenuated I/R-induced increases in oxidative stress, autophagic flux, and Beclin-1 expression, accompanied by a decrease in the size of myocardial infarction (MI)/area at risk (AAR), suggesting that oxidative stress plays an important role in mediating autophagy and myocardial injury during I/R. MI/AAR after I/R was significantly reduced in beclin1(+/-) mice, whereas beclin1(+/-) mice treated with MPG exhibited no additional reduction in the size of MI/AAR after I/R. These results suggest that oxidative stress plays an important role in mediating autophagy during I/R, and that activation of autophagy through oxidative stress mediates myocardial injury in response to I/R in the mouse heart.


Predictors of positive surgical margins at open and robot-assisted laparoscopic radical prostatectomy: a single surgeon series.
Robot-assisted laparoscopic radical prostatectomy (RALRP), increasingly used to treat localized prostate cancer, has advantages over open radical prostatectomy (ORP) in terms of reduced bleeding and quicker convalescence. However, debate continues over whether RALRP provides superior or at least equivalent surgical outcomes. This study compares positive surgical margins (+SM), as a surrogate for long-term cancer control, at RALRP and ORP performed by a single experienced surgeon during the process of taking up RALRP. 400 consecutive patients undergoing surgery for prostate cancer under a single surgeon (DW) between November 1999 and July 2009 were studied. Prior to July 2005, all patients underwent ORP; after this date, most patients were treated by RALRP. Data were collected by retrospective chart review and analysed independently of the treating surgeon. +SM were defined as the presence of cancer at an inked surface. Overall, 23 (11.5%) of 200 patients undergoing RALRP had +SM, compared to 40 (20.0%) of 200 patients undergoing ORP (P < 0.05). On univariate logistic regression analysis, in addition to surgical approach (odds ratio [OR] = 1.92), patient age (OR = 1.05), pathologic stage (OR = 3.93) and specimen Gleason (GS) score (OR = 1.86) were significant predictors of +SM. On multivariate analysis, surgical approach, p-stage and specimen GS remained significant predictors of +SM. RALRP is associated with lower rates of +SM compared to ORP, even after adjusting for other known risk factors. Of note, the RALRP in this study were part of the surgeon's learning curve.


Antibody responses to SARS-CoV-2 Omicron infection in patients with hematological malignancies: A multicenter, prospective cohort study.
Little is known about antibody responses to natural Omicron infection and the risk factors for poor responders in patients with hematological malignancies (HM). We conducted a multicenter, prospective cohort study during the latest Omicron wave in Chongqing, China, aiming to compare the antibody responses, as assessed by IgG levels of anti-receptor binding domain of spike protein (anti-S-RBD), to Omicron infection in the HM cohort (HMC) with healthy control cohort (HCC), and solid cancer cohort (SCC). In addition, we intend to explore the risk factors for poor responders in the HMC. Among the 466 HM patients in this cohort, the seroconversion rate was 92.7%, no statistically difference compared with HCC (98.2%, p = 0.0513) or SCC (100%, p = 0.1363). The median anti-S-RBD IgG titer was 29.9 ng/mL, significantly lower than that of HCC (46.9 ng/mL, p < 0.0001) or SCC (46.2 ng/mL, p < 0.0001). Risk factors associated with nonseroconversion included no COVID-19 vaccination history (odds ratio [OR] = 4.58, 95% confidence interval [CI]: 1.75-12.00, p = 0.002), clinical course of COVID-19 ≤ 7 days (OR = 2.86, 95% CI: 1.31-6.25, p = 0.008) and severe B-cell reduction (0-10/μL) (OR = 3.22, 95% CI: 1.32-7.88, p = 0.010). Risk factors associated with low anti-S-RBD IgG titer were clinical course of COVID-19 ≤ 7 days (OR = 2.58, 95% CI: 1.59-4.18, p < 0.001) and severe B-cell reduction (0-10/μL) (OR = 2.87, 95% CI: 1.57-5.24, p < 0.001). This study reveals a poor antibody responses to Omicron (BA.5.2.48) infection in HM patients and identified risk factors for poor responders. Highlights that HM patients, especially those with these risk factors, may be susceptible to SARS-CoV-2 reinfection, and the postinfection vaccination strategies for these patients should be tailored. Clinical trial: ChiCTR2300071830.


Cybertutor: a teaching tool in Dermatology.
It was developed a teaching tool in Dermatology for undergraduate medical students, using an interactive website, the Cybertutor. Clinical cases, lectures and updated bibliography were selected. Photographies of dermatological lesions were taken from ambulatory patients. The topics of the lectures were based on the current curriculum of the Federal University of Rio Grande do Sul. The Cybertutor is a dynamic and modern teaching tool, allowing constant innovation.


Variable myopathic presentation in a single family with novel skeletal RYR1 mutation.
We describe an autosomal recessive heterogeneous congenital myopathy in a large consanguineous family. The disease is characterized by variable severity, progressive course in 3 of 4 patients, myopathic face without ophthalmoplegia and proximal muscle weakness. Absence of cores was noted in all patients. Genome wide linkage analysis revealed a single locus on chromosome 19q13 with Zmax = 3.86 at θ = 0.0 and homozygosity of the polymorphic markers at this locus in patients. Direct sequencing of the main candidate gene within the candidate region, RYR1, was performed. A novel homozygous A to G nucleotide substitution (p.Y3016C) within exon 60 of the RYR1 gene was found in patients. ARMS PCR was used to screen for the mutation in all available family members and in an additional 150 healthy individuals. This procedure confirmed sequence analysis and did not reveal the A to G mutation (p.Y3016C) in 300 chromosomes from healthy individuals. Functional analysis on EBV immortalized cell lines showed no effect of the mutation on RyR1 pharmacological activation or the content of intracellular Ca(2+) stores. Western blot analysis demonstrated a significant reduction of the RyR1 protein in the patient's muscle concomitant with a reduction of the DHPRα1.1 protein. This novel mutation resulting in RyR1 protein decrease causes heterogeneous clinical presentation, including slow progression course and absence of centrally localized cores on muscle biopsy. We suggest that RYR1 related myopathy should be considered in a wide variety of clinical and pathological presentation in childhood myopathies.


A ReaxFF molecular dynamics study of molecular-level interactions during binder jetting 3D-printing.
In the present work, we study one of the major additive manufacturing processes, i.e., the binder jetting printing (BJP) process, at the molecular level through atomistic-scale level representations of powders and binder solutions with chromium-oxide (Cr-oxide) nanoparticles and water-based diethylene glycol solutions, respectively. The results show that both diethylene glycol and water contribute to the bonding of Cr-oxide particles during the print and curing stages by forming a hydrogen bond network. Heating the system to the burn-out temperature results in the oxidation of diethylene glycol and the decomposition of the hydrogen bond network. Subsequently, Cr-oxide particles are partially sintered by forming Cr-O bonds. The final sintering facilitates further Cr-O bond formation. Additionally, the influence of the chemical composition of the binder solution is investigated by performing ReaxFF molecular dynamics simulations on two sets of systems, which control the number of water and diethylene glycol molecules, respectively. Our results demonstrate that adding both diethylene glycol and water to the binder solution can raise the number of "useful" hydrogen bonds, resulting in a higher breaking strength at the print and curing stages. During the burn-out and sintering stages, the influence of water on the breaking strength is not obvious. In contrast, an optimal quantity of binder species exists for the breaking strength after sintering. A comparison of the ReaxFF molecular dynamics simulations using 2-ethoxyethanol, diethylene glycol and 1-(2,2,2-trihydroxyethoxy)ethane-2,2,2-triol as the binder phase indicates that an increasing number of hydroxyl groups leads to higher breaking strength at the print and curing stages. The findings from this study can be extended to identify the optimal binder chemistry, curing and sintering conditions for different material systems.


Acute Care Referral Systems in Liberia: Transfer and Referral Capabilities in a Low-Income Country.
Introduction Following two decades of armed conflict in Liberia, over 95% of health care facilities were partially or completely destroyed. Although the Liberian health system has undergone significant rehabilitation, one particular weakness is the lack of organized systems for referral and prehospital care. Acute care referral systems are a critical component of effective health care delivery and have led to improved quality of care and patient outcomes. Problem This study aimed to characterize the referral and transfer systems in the largest county of Liberia. A cross-sectional, health referral survey of a representative sample of health facilities in Montserrado County, Liberia was performed. A systematic random sample of all primary health care (PHC) clinics, fraction proportional to district population size, and all secondary and tertiary health facilities were included in the study sample. Collected data included baseline information about the health facility, patient flow, and qualitative and quantitative data regarding referral practices. A total of 62 health facilities-41 PHC clinics, 11 health centers (HCs), and 10 referral hospitals (RHs)-were surveyed during the 6-week study period. In sum, three percent of patients were referred to a higher-level of care. Communication between health facilities was largely unsystematic, with lack of specific protocols (n=3; 5.0%) and standardized documentation (n=26; 44.0%) for referral. While most health facilities reported walking as the primary means by which patients presented to initial health facilities (n=50; 81.0%), private vehicles, including commercial taxis (n=37; 60.0%), were the primary transport mechanism for referral of patients between health facilities. This study identified several weaknesses in acute care referral systems in Liberia, including lack of systematic care protocols for transfer, documentation, communication, and transport. However, several informal, well-functioning mechanisms for referral exist and could serve as the basis for a more robust system. Well-integrated acute care referral systems in low-income countries, like Liberia, may help to mitigate future public health crises by augmenting a country's capacity for emergency preparedness. Kim J , Barreix M , Babcock C , Bills CB . Acute care referral systems in Liberia: transfer and referral capabilities in a low-income country. Prehosp Disaster Med. 2017;32(6):642-650.


[Pharmacological properties of pteleprenine, a quinoline alkaloid from Orixa japonica (Rutaceae), in guinea pig ileum and canine left atrium].
We investigated the pharmacological properties of pteleprenine, a quinoline alkaloid contained in the Rutaceous plant, Orixa japonica, on the contractile responses of the guinea pig ileum and on the inotropic responses of the canine left atrium. Contractile responses of the ileum to acetylcholine and histamine were not inhibited by less than 10(-6) M of pteleprenine. Meanwhile, the nicotine induced-contraction of the ileum was dose-dependently diminished by pteleprenine, but the contractile response to nicotine did not reach the maximum value in the presence of 10(-5) M of pteleprenine. The pA2 value of pteleprenine was 6.6 as determined from the Schild plot, which slope was nearly unity. Furthermore, the contraction of the ileum by 10(-5) M of 1,1-dimethyl-4-phenylpiperazinium (DMPP), a specific agonist of nicotinic acetylcholine receptors, was also dose-dependently suppressed by pteleprenine. On the other hand, 10(-5) M of pteleprenine did not have considerable inhibitory effects on acetylcholine- and nicotine-induced negative inotropic effects in the canine left atrium. From these results, it is suggested that pteleprenine has a specific inhibitory effect on nicotinic acetylcholine receptors in the guinea pig ileum.


A Novel Automatic Digital Algorithm that Accurately Quantifies Steatosis in NAFLD on Histopathological Whole-Slide Images.
Accurate assessment of hepatic steatosis is a key to grade disease severity in non-alcoholic fatty liver disease (NAFLD). We developed a digital automated quantification of steatosis on whole-slide images (WSIs) of liver tissue and performed a validation study. Hematoxylin-eosin stained liver tissue slides were digitally scanned, and steatotic areas were manually annotated. We identified thresholds for size and roundness parameters by logistic regression to discriminate steatosis from surrounding liver tissue. The resulting algorithm produces a steatosis proportionate area (SPA; ratio of steatotic area to total tissue area described as percentage). The software can be implemented as a Java plug-in in FIJI, in which digital WSI can be processed automatically using the Pathomation extension. We obtained liver tissue specimens from 61 NAFLD patients and 18 controls. The area under the curve of correctly classified steatosis by the algorithm was 0.970 (95% CI 0.968-0.973), P < 0.001. Accuracy of the algorithm was 91.9%, with a classification error of 8.1%. SPA correlated significantly with steatosis grade (Rs = 0.845, CI: 0.749-0.902, P < 0.001) and increased significantly with each individual steatosis grade, except between Grade 2 and 3. We have developed a novel digital analysis algorithm that accurately quantifies steatosis on WSIs of liver tissue. This algorithm can be incorporated when quantification of steatosis is warranted, such as in clinical trials studying efficacy of new therapeutic interventions in NAFLD. © 2019 The Authors. Cytometry Part B: Clinical Cytometry published by Wiley Periodicals, Inc. on behalf of International Clinical Cytometry Society.


Potential Application of Self-Assembled Peptides and Proteins in Breast Cancer and Cervical Cancer.
Ongoing research is gradually broadening the idea of cancer treatment, with attention being focused on nanoparticles to improve the stability, therapeutic efficacy, targeting, and other important metrics of conventional drugs and traditional drug delivery methods. Studies have demonstrated that drug delivery carriers based on biomaterials (e.g., protein nanoparticles and lipids) and inorganic materials (e.g., metal nanoparticles) have potential anticancer effects. Among these carriers, self-assembled proteins and peptides, which are highly biocompatible and easy to standardize and produce, are strong candidates for the preparation of anticancer drugs. Breast cancer (BC) and cervical cancer (CC) are two of the most common and deadly cancers in women. These cancers not only threaten lives globally but also put a heavy burden on the healthcare system. Despite advances in medical care, the incidence of these two cancers, particularly CC, which is almost entirely preventable, continues to rise, and the mortality rate remains steady. Therefore, there is still a need for in-depth research on these two cancers to develop more targeted, efficacious, and safe therapies. This paper reviews the types of self-assembling proteins and peptides (e.g., ferritin, albumin, and virus-like particles) and natural products (e.g., soy and paclitaxel) commonly used in the treatment of BC and CC and describes the types of drugs that can be delivered using self-assembling proteins and peptides as carriers (e.g., siRNAs, DNA, plasmids, and mRNAs). The mechanisms (including self-assembly) by which the natural products act on CC and BC are discussed. The mechanism of action of natural products on CC and BC and the mechanism of action of self-assembled proteins and peptides have many similarities (e.g., NF-KB and Wnt). Thus, natural products using self-assembled proteins and peptides as carriers show potential for the treatment of BC and CC.


Analysis of neuroblastoma tumour progression; loss of PHOX2B on 4p13 and 17q gain are early events in neuroblastoma tumourigenesis.
Neuroblastomas are biologically and clinically heterogeneous tumours that most often occur sporadically in children at median age of 2 years. The PHOX2B gene is implicated in the development of the autonomic nervous system and has been found to be infrequently mutated in sporadic neuroblastoma tumours and in some patients with hereditary neuroblastoma. We have screened a selected series of 36 paediatric tumours with presumed genetic predisposition, 34 of them neuroblastomas, for mutations in PHOX2B. A constitutional heterozygous missense mutation was found in a boy who developed bilateral adrenal tumours and stage 4 disease during infancy. The second allele of the PHOX2B locus was lost in the tumour DNA. Histopathological evaluation of the tumours suggested growth of two primary tumours, one with diploid DNA content and the other with tetraploid DNA content, i.e. a case of neuroblastoma stage 4M (multifocal tumour). However, array CGH (comparative genomic hybridization) data performed on both tumour masses from the patient instead supported a model where a common malignant precursor gave rise to the diploid tumour and subsequently the tetraploid tumour have progressed from the common precursor or by metastasis from the diploid tumour with additional genetic changes. The whole genome dosage analysis showed that the remaining alleles of PHOX2B had been lost in both tumours together with a specific 17q gain pattern. The tetraploid tumour had these features together with additional whole chromosomal loss of chromosomes 3, 9, 14 and 15. Based on the data presented here we suggest that loss of PHOX2B and 17q gain are early events in neuroblastoma tumourigenesis. We also propose investigators to re-analyze the rare cases of multifocal neuroblastomas with the array CGH technique for better understanding of the origin of these tumours.


In situ transformation of hydraulic fracturing surfactants from well injection to produced water.
Chemical changes to hydraulic fracturing fluids (HFFs) within fractured unconventional reservoirs may affect hydrocarbon recovery and, in turn, the environmental impact of unconventional oil and gas development. Ethoxylated alcohol surfactants, which include alkyl ethoxylates (AEOs) and polyethylene glycols (PEGs), are often present in HFF as solvents, non-emulsifiers, and corrosion inhibitors. We present detailed analysis of polyethoxylates in HFF at the time of injection into three hydraulically fractured Marcellus Shale wells and in the produced water returning to the surface. Despite the addition of AEOs to the injection fluid during almost all stages, they were rarely detected in the produced water. Conversely, while PEGs were nearly absent in the injection fluid, they were the dominant constituents in the produced water. Similar numbers of ethoxylate units support downhole transformation of AEOs to PEGs through central cleavage of the ethoxylate chain from the alkyl group. We also observed a decrease in the average ethoxylate (EO) number of the PEG-EOs in the produced water over time, consistent with biodegradation during production. Our results elucidate an overlooked surfactant transformation pathway that may affect the efficacy of HFF to maximize oil and gas recovery from unconventional shale reservoirs.


Color constancy: a method for recovering surface spectral reflectance.
Human and machine visual sensing is enhanced when surface properties of objects in scenes, including color, can be reliably estimated despite changes in the ambient lighting conditions. We describe a computational method for estimating surface spectral reflectance when the spectral power distribution of the ambient light is not known.


Eggerthella lenta bacteraemia in endometrial adenocarcinoma - a case report from Pakistan.
We report a case of Eggerthella lenta bacteraemia. An elderly lady with metastasised endometrial adenocarcinoma presented with complaints of fever nausea vomiting and abdominal pain. CT scan of the abdomen showed enlarged liver with multiple metastatic lesions raising suspicion of small-bowel obstruction. Due to multiple comorbid conditions, surgery was contraindicated and she was treated empirically with meropenem and vancomycin. Blood culture received on admission grew Eggerthela lenta.


Adherence and persistence among chronic myeloid leukemia patients during second-line tyrosine kinase inhibitor treatment.
Chronic myeloid leukemia (CML) treatment is lifelong, and while it is important for patients to remain adherent to treatment, there are conflicting findings with respect to differences in adherence and persistence with dasatinib or nilotinib during second-line treatment. To compare the rates of adherence, persistence, and discontinuation of 2 oral second-generation tyrosine kinase inhibitors (TKI), dasatinib and nilotinib, in CML patients during their first 12 months of second-line treatment. Adult CML patients (ICD-9-CM: 205.1x) with 2 consecutive dasatinib or nilotinib prescription claims within 12 months were identified from the Truven Health MarketScan Databases (January 1, 2006-September 30, 2011). Patients were excluded if they had FDA-approved non-CML indications for imatinib, had less than 6 months continuous enrollment, or had a stem cell/bone marrow transplant in the 6 months pre-index. Patients were followed until the first occurrence of index TKI discontinuation/switch; enrollment end; December 31, 2011; or 12 months follow-up post-index. Index treatment (dasatinib ≤ 100 mg or nilotinib) was categorized as second-line if there was evidence of only 1 alternative TKI exposure (e.g., imatinib, dasatinib, or nilotinib) anytime during the patient's available claims history. When comparing adherence, persistence, and discontinuation, inverse probability treatment weighting (IPTW) was used. Adherence and persistence measures were calculated as specified by the International Society for Pharmacoeconomics and Outcomes Research Medication Compliance and Persistence Special Interest Group. Treatment adherence was calculated using medication possession ratio (MPR) and was reported as both continuous and binary measures (i.e., high adherence = MPR ≥ 85%). Persistence was reported as the proportion of days covered (PDC) and estimated level of persistence (ELPT). Finally, discontinuation was defined as a treatment gap of greater than 90 days and absence of index TKI during the remainder of the follow-up period. Time to discontinuation and high adherence of index TKI were compared using weighted Cox proportional hazards and logistic regression models, respectively. After propensity weighting, the 219 second-line dasatinib patients and the 158 second-line nilotinib patients were similar in mean age, gender, cancer complexity, and comorbidity burden at baseline. Age as a categorical measure, population density, and index year remained imbalanced and were, therefore, included as covariates in the multivariate analysis of adherence. In the bivariate analyses, mean MPR (88.2% vs. 84.4%, P = 0.036); proportion of patients with high adherence (72.7% vs. 63.3%, P = 0.006); and ELPT (70.4% vs. 62.7%, P = 0.026) were significantly higher among dasatinib patients than nilotinib patients. Mean PDC was not significantly different between dasatinib and nilotinib patients (0.79 vs. 0.77, P = 0.328) after propensity weighting. In addition, a significantly lower proportion of second-line dasatinib patients discontinued their index therapy compared with second-line nilotinib patients (4.4% vs. 8.6%, P = 0.020). With a hazard ratio (HR) of 0.50 (95% CI = 0.27-0.93, P = 0.029), dasatinib patients had half the possibility of discontinuing treatment compared with nilotinib patients at any point in time. After accounting for the baseline factors remaining imbalanced and controlling for cancer complexity and number of concomitant medications at baseline, second-line dasatinib patients were 1.7 times (95% CI = 1.2-2.4) more likely to be highly adherent than second-line nilotinib patients (P = 0.0016). Among second-line TKI-treated CML patients, dasatinib patients had significantly higher adherence and lower discontinuation rates compared with patients receiving second-line nilotinib.


Frequency and timing of coital orgasm in women desirous of becoming pregnant.
Generally, no functional role, except pleasure, is assigned to a women's coital orgasm. Recently, however, researchers have suggested that women can regulate the number of sperm accepted or rejected by manipulation coital orgasm timing. Baker and Bellis (1995) demonstrated that women retain more sperm if coital orgasm occurs after, rather than before male ejaculation. The present study investigated (i) factors associated with coital orgasm frequency and (ii) whether women desirous of becoming pregnant report more coital orgasms after than before their partners. Sixty-nine women rated their desire for pregnancy and answered questions about their sexual behaviors, coital orgasm frequency, and coital orgasm timing in relation to their partners. In regression analysis, coital orgasm frequency was predicted by foreplay duration, masturbation frequency, and active sexual participation; desire for pregnancy did not predict coital orgasm frequency. Desire for pregnancy was predicted by age (negatively), active sexual participation (positively), and positively by orgasm after partner ejaculation. Consistent with the Baker and Bellis hypothesis, frequency of orgasm after partner's orgasm remained a significant predictor for desire for pregnancy after controlling for degree of marital happiness, sexual foreplay duration, and coital orgasm frequency.


Global analysis of depletion and recovery of seabed biota after bottom trawling disturbance.
Bottom trawling is the most widespread human activity affecting seabed habitats. Here, we collate all available data for experimental and comparative studies of trawling impacts on whole communities of seabed macroinvertebrates on sedimentary habitats and develop widely applicable methods to estimate depletion and recovery rates of biota after trawling. Depletion of biota and trawl penetration into the seabed are highly correlated. Otter trawls caused the least depletion, removing 6% of biota per pass and penetrating the seabed on average down to 2.4 cm, whereas hydraulic dredges caused the most depletion, removing 41% of biota and penetrating the seabed on average 16.1 cm. Median recovery times posttrawling (from 50 to 95% of unimpacted biomass) ranged between 1.9 and 6.4 y. By accounting for the effects of penetration depth, environmental variation, and uncertainty, the models explained much of the variability of depletion and recovery estimates from single studies. Coupled with large-scale, high-resolution maps of trawling frequency and habitat, our estimates of depletion and recovery rates enable the assessment of trawling impacts on unprecedented spatial scales.


Activity of amoxycillin against enterococci and synergism with aminoglycoside antibiotics.
Amoxycillin was more active in vitro than ampicillin or benzylpenicillin against clinical isolates of enterococci. All 55 strains tested were sensitive to the three penicillins but 15 strains showed a high level of resistance to streptomycin and two of these were also insensitive to kanamycin. All strains were sensitive to gentamicin, which was the most active of the aminoglycoside antibiotics. The penicillins showed pronounced bactericidal activity against the enterococci but failed to sterilise cultures of these organisms. Combinations of penicillins and aminoglycosides invariably produced synergistic bactericidal effects which resulted in sterilisation of cultures of these organisms. Combinations of penicillins and aminoglycosides invariably produced synergistic bactericidal effects which resulted in sterilisation of cultures of enterococci provided that the strain was sensitive to the aminoglycoside moiety of the antibiotic combination. Synergism was not observed with a combination of a penicillin and an aminoglycoside when the enterococcus was resistant to the aminoglycoside. The data reported suggest that amoxycillin may have certain advantages, compared with ampicillin or benzylpenicillin, for the treatment of enterococcal infections.


Positional specificity of a phospholipase A activity induced by wounding, systemin, and oligosaccharide elicitors in tomato leaves
Phospholipase A (PLA) activity, as measured by the accumulation of (14)C-lysophosphatidylcholine in leaves of tomato plants, increased rapidly and systemically in response to wounding. The increase in PLA activity in the systemic unwounded leaves was biphasic in wild-type tomato plants, peaking at 15 min and again at 60 min, but the second peak of activity was absent in transgenic prosystemin antisense plants. Supplying young excised tomato plants with the polypeptide hormone systemin also caused (14)C-lysophosphatidylcholine to increase to levels similar to those induced by wounding, but the increase in activity persisted for >2 hr. Antagonists of systemin blocked both the release of (14)C-lysophosphatidylcholine and the accumulation of defense proteins in response to systemin. (14)C-lysophosphatidylcholine levels did not increase in response to jasmonic acid. Chemical acylation of the lysophosphatidylcholine produced by wounding, systemin, and oligosaccharide elicitors followed by enzymatic hydrolysis with lipases of known specificities demostrated that the lysophosphatidylcholine is generated by a PLA with specificity for the sn-2 position.


Construction and characterization of an Escherichia coli strain with a uncI mutation.
A strain of Escherichia coli with a mutation in the promoter proximal gene ( uncI ) of the unc operon has been constructed by using a new gene replacement method. The mutation is a deletion of a defined sequence of 196 base pairs. It was constructed by homologous integration and segregation of a ColE1-derived recombinant plasmid containing the mutation, in a temperature-sensitive polA strain. The mutant strain is phenotypically unc+ but has a reduced growth yield compared to a normal sibling strain.


Differential induction of functional B1-bradykinin receptors along the rat nephron in endotoxin induced inflammation.
Under physiological conditions, the effects of kinins in the kidney are mainly mediated by the bradykinin B2-receptor, whereas the kinin B1-receptor is strongly induced under inflammatory conditions in a variety of tissues. Knowledge of the distribution of the B1-receptor along the nephron is of importance since the B1-receptor might replace B2-receptors under these conditions. Using a RT-PCR/Southern blot approach allowing relative quantification of mRNA levels, ten different microdissected rat nephron segments were analyzed for the presence of the B1- and B2-receptor before and after endotoxin treatment to induce experimental inflammation. The functionality of the expressed receptors was assessed by kinin-induced intracellular calcium ([Ca2+]i) mobilization in microdissected nephron segments. While under physiological conditions no B1-receptor mRNA could be detected, after 18 hours of treatment with bacterial lipopolysaccharide (LPS) the expression of B1-receptor mRNA was strongly induced in the efferent arteriole, the medullary and inner medullary thin limb, and in the distal tubule. Moderate expression was found in the glomerulus, proximal convoluted and straight tubules, and in the medullary thick ascending limb. Small but detectable expression was observed in the cortical collecting duct. The induction of B1-receptor mRNA expression resulted in functional receptor expression, since increases in [Ca2+]i were observed upon B1-agonist stimulation. LPS treatment also increased the expression of B2-receptor mRNA in all nephron segments except in the glomerulus, the inner medullary thin limb and the outer medullary collecting duct. However, no related changes in B2-agonist induced rises in [Ca2+]i were found. These studies show a functional induction of the B1-kinin receptor along the rat nephron, which should be taken in account to address the effects of kinins under inflammatory conditions in the kidney.


Characterization of integrative and conjugative element ICEKp1-associated genomic heterogeneity in a Klebsiella pneumoniae strain isolated from a primary liver abscess.
Genomic heterogeneity has been shown to be associated with Klebsiella pneumoniae strains causing pyogenic liver abscesses (PLA) and metastatic infections. In order to explore the mechanism responsible for genomic heterogeneity in K. pneumoniae, we compared the complete genomic sequences of strains NTUH-K2044 and MGH78578. An approximately 76-kbp DNA fragment located adjacent to an asparagine (asn) tRNA gene was present in NTUH-K2044 but not in MGH78578. This fragment could be divided into three regions with different functions, and structurally it resembled a functional integrative and conjugative element (ICE), ICEEc1, in Escherichia coli. The 5' region of this fragment contained genes similar to a high-pathogenicity island (HPI) of Yersinia pestis and Yersinia pseudotuberculosis. The middle region was similar to part of a large plasmid in K. pneumoniae, and the 3' region contained genes responsible for DNA conjugative transfer. Therefore, this DNA fragment was designated ICEKp1. Precise excision and extrachromosomal circularization of ICEKp1 were detected in K. pneumoniae wild-type strain NTUH-K2044. ICEKp1 could integrate into the asn tRNA loci of the chromosome of another K. pneumoniae isolate. The prevalence of ICEKp1 was higher in PLA strains (38 of 42 strains) than in non-tissue-invasive strains (5 of 32 strains). Therefore, ICEKp1 may contribute to the transmission of the HPI and result in K. pneumoniae PLA infection-associated genomic heterogeneity.


Apoptotic Bax at Oxidatively Stressed Mitochondrial Membranes: Lipid Dynamics and Permeabilization.
Mitochondria are crucial compartments of eukaryotic cells because they function as the cellular power plant and play a central role in the early stages of programmed cell death (apoptosis). To avoid undesired cell death, this apoptotic pathway is tightly regulated by members of the Bcl-2 protein family, which interact on the external surface of the mitochondria, i.e., the mitochondrial outer membrane (MOM), and modulate its permeability to apoptotic factors, controlling their release into the cytosol. A growing body of evidence suggests that the MOM lipids play active roles in this permeabilization process. In particular, oxidized phospholipids (OxPls) formed under intracellular stress seem to directly induce apoptotic activity at the MOM. Here we show that the process of MOM pore formation is sensitive to the type of OxPls species that are generated. We created MOM-mimicking liposome systems, which resemble the cellular situation before apoptosis and upon triggering of oxidative stress conditions. These vesicles were studied using 31P solid-state magic-angle-spinning nuclear magnetic resonance spectroscopy and differential scanning calorimetry, together with dye leakage assays. Direct polarization and cross-polarization nuclear magnetic resonance experiments enabled us to probe the heterogeneity of these membranes and their associated molecular dynamics. The addition of apoptotic Bax protein to OxPls-containing vesicles drastically changed the membranes' dynamic behavior, almost completely negating the previously observed effect of temperature on the lipids' molecular dynamics and inducing an ordering effect that led to more cooperative membrane melting. Our results support the hypothesis that the mitochondrion-specific lipid cardiolipin functions as a first contact site for Bax during its translocation to the MOM in the onset of apoptosis. In addition, dye leakage assays revealed that different OxPls species in the MOM-mimicking vesicles can have opposing effects on Bax pore formation.


Synthesis and testing of 5-benzyl-2,4-diaminopyrimidines as potential inhibitors of leishmanial and trypanosomal dihydrofolate reductase.
Dihydrofolate reductase is a drug target that has not been thoroughly investigated in leishmania and trypanosomes. Work has previously shown that 5-benzyl-2,4-diaminopyrimidines are selective inhibitors of the leishmanial and trypanosome enzymes. Modelling predicted that alkyl/aryl substitution on the 6-position of the pyrimidine ring should increase enzyme activity of 5-benzyl-2,4-diaminopyrimidines as inhibitors of leishmanial and trypanosomal dihydrofolate reductase. Various compounds were prepared and evaluated against both the recombinant enzymes and the intact organisms. The presence of a substituent had a small or negative effect on activity against the enzyme or intact parasites compared to unsubstituted compounds.


[Study of Navarra (PECNA). Correlation of arterial blood pressure, in a child-young population, with anthropometric and biochemical parameters].
As part of an epidemiological study on cardiovascular risk factors among children and adolescent in Navarra, arterial blood pressure (BP) readings were taken in 5,829 children. These children, both males and females, between the ages of 4 and 17 years, were selected at random from the public and private school population in our community. The correlation between systolic blood pressure and diastolic blood pressure with anthropometric and biochemical parameters was analyzed. The correlation coefficient for systolic BP readings were always higher than those for diastolic BP. The highest correlations for both BP scores were found with weight, followed by height. Correlation with total weight parameters is superior to that found with body fat parameters. There were no significant correlations between BP and the lipid parameters. When analyzing multiple regression equations, we find that with only the child's weight and age, both systolic and diastolic BP can be predicted with correlation coefficients of 0.597 and 0.492, respectively. When doing a partial correlation analysis, the correlation between systolic BP and age, at a fixed height value, disappears; while the correlation of systolic BP with height remains when using a set age. This suggest that the best definition for hypertension is based on BP-height percentile, rather than BP-age.


In vitro inhibition of ocular cell proliferation with ara-C: blockage of the antiproliferative effect with 2'-deoxycytidine.
Cytosine arabinoside (ara-C), a cell cycle-phase specific antimetabolite, was tested in a tissue culture model of human scleral fibroblasts (HSCF), and human retinal pigmented epithelium cells (HRPE), to investigate possible applications in the modulation of wound healing after glaucoma filtering surgery and in the postoperative management of proliferative vitreoretinopathy. Cell proliferation after a 72 hr incubation in the presence of ara-C was inhibited to 50% of the control growth rate (ID50) with calculated doses of 60 ng/ml in HSCF and 4 and 5 ng/ml in HRPE cultures. 2'-deoxycytidine, a competitive inhibitor of ara-C action, completely eliminated the cell growth retardation of both cell types produced by ara-C after 72 hr incubation.


Purification, enzymatic characterization, and nucleotide sequence of a high-isoelectric-point alpha-glucosidase from barley malt.
High-isoelectric-point (pI) alpha-glucosidase was purified 7, 300-fold from an extract of barley (Hordeum vulgare) malt by ammonium sulfate fractionation, ion-exchange, and butyl-Sepharose chromatography. The enzyme had high activity toward maltose (k(cat) = 25 s(-1)), with an optimum at pH 4.5, and catalyzed the hydrolysis by a retaining mechanism, as shown by nuclear magnetic resonance. Acarbose was a strong inhibitor (K(i) = 1.5 microM). Molecular recognition revealed that all OH-groups in the non-reducing ring and OH-3 in the reducing ring of maltose formed important hydrogen bonds to the enzyme in the transition state complex. Mass spectrometry of tryptic fragments assigned the 92-kD protein to a barley cDNA (GenBank accession no. U22450) that appears to encode an alpha-glucosidase. A corresponding sequence (HvAgl97; GenBank accession no. AF118226) was isolated from a genomic phage library using a cDNA fragment from a barley cDNA library. HvAgl97 encodes a putative 96.6-kD protein of 879 amino acids with 93.8% identity to the protein deduced from U22450. The sequence contains two active site motifs of glycoside hydrolase family 31. Three introns of 86 to 4,286 bp interrupt the coding region. The four exons vary from 218 to 1,529 bp. Gene expression analysis showed that transcription reached a maximum 48 h after the start of germination.


Improving influenza vaccination rates for high-risk inpatients.
In 1987, the Minneapolis Veterans Affairs Medical Center implemented an ongoing, multifaceted influenza vaccination program that targeted all high-risk outpatients followed at the medical center. While the program achieved and sustained vaccination rates of 60% or more for high-risk outpatients, vaccination rates for high-risk inpatients continued to be 25% or less. Beginning with the 1989-1990 immunization season, the "flu shot" program was extended to include all high-risk inpatients. Both the outpatient and inpatient components of the Minneapolis Flu Shot Program emphasized administrative and organizational elements. The program's goal was to achieve vaccination rates of at least 60% for both high-risk outpatients and inpatients. This study assesses the effectiveness of the inpatient program among elderly patients hospitalized on the medical service during the immunization season. To estimate patient risk characteristics and vaccination rates, 50% of elderly patients who were discharged from the medical service during the immunization season were surveyed using a validated postcard questionnaire. Their responses were compared with the responses of elderly patients to the annual outpatient survey conducted to assess the "flu shot" program. Overall, 78.6% of elderly respondents discharged from the medical service during the immunization season were vaccinated. Vaccination rates within various high-risk subgroups all exceeded 70%. None of these vaccination rates for inpatient groups differed significantly from the rates for corresponding outpatients. An outpatient "flu shot" program that emphasizes administrative and organizational elements can be successfully expanded to high-risk inpatients. The vaccination rates attained with such a program may not only achieve but exceed the national health objective for influenza vaccination.


Diagnostic efficacy of the triglyceride-glucose index in the prediction of contrast-induced nephropathy following percutaneous coronary intervention.
Contrast-induced nephropathy (CIN) is a common complication of percutaneous coronary intervention (PCI). Identifying patients at high CIN risk remains challenging. The triglyceride-glucose (TyG) index may help predict CIN but evidence is limited. We conducted a meta-analysis to evaluate the diagnostic value of TyG index for CIN after PCI. A systematic literature search was performed in MEDLINE, Cochrane, and EMBASE until August 2023 (PROSPERO registration: CRD42023452257). Observational studies examining TyG index for predicting CIN risk in PCI patients were included. This diagnostic meta-analysis aimed to evaluate the accuracy of the TyG index in predicting the likelihood of CIN. Secondary outcomes aimed to assess the pooled incidence of CIN and the association between an elevated TyG index and the risk of CIN. Five studies (Turkey, n=2; China, n=3) with 3518 patients (age range: 57.6 to 68.22 years) were included. The pooled incidence of CIN was 15.3% [95% confidence interval (CI) 11-20.8%]. A high TyG index associated with increased CIN risk (odds ratio: 2.25, 95% CI 1.82-2.77). Pooled sensitivity and specificity were 0.77 (95% CI 0.59-0.88) and 0.55 (95% CI 0.43-0.68) respectively. Analysis of the summary receiver operating characteristic (sROC) curve revealed an area under the curve of 0.69 (95% CI 0.65-0.73). There was a low risk of publication bias (p = 0.81). The TyG index displayed a noteworthy correlation with the risk of CIN subsequent to PCI. However, its overall diagnostic accuracy was found to be moderate in nature. While promising, the TyG index should not be used in isolation for CIN screening given the heterogeneity between studies. In addition, the findings cannot be considered conclusive given the scarcity of data. Further large-scale studies are warranted to validate TyG cutoffs and determine how to optimally incorporate it into current risk prediction models. https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42023452257, identifier CRD42023452257.


[Prophylactic medicine--a foreground direction of medical support of the air staff of aviation of the Armed Forces of Russian Federation].
The first prophylaxis of chronic not-infection diseases obtains more and more value. Level of these diseases among air staff is rather high and has a tendency of increasing. It's bounded up with defects in the system of the first medical unit, insufficient conducting of ecological and sanitarium-hygiene screening and taking steps on decreasing of the influence of noxious agent on the human organism, improvement of working conditions of air specialists, decreasing of the value of psychoemotional stress and other factors. An important factor during the conducting the first and the second prophylaxis is an early detection of changes in health by the air staff and technical-engineer staff, diagnostics of diseases on early stages, when they conduct without any symptoms, forehanded rehabilitation and treatment.


CD133 antibody-conjugated immunoliposomes encapsulating gemcitabine for targeting glioblastoma stem cells.
CD133+ cells in glioblastoma multiforme (GBM) display glioma stem cell (GSC) properties and have been considered to be the culprit in tumor recurrence, justifying the exploration of potential therapeutic modalities targeting CD133+ cells. Such strategies require a drug-delivery vehicle, and increasingly, the unique properties of PEGylated liposomes are being exploited for this purpose. More advanced liposomal delivery studies have suggested conjugation of CD133 antibodies as a suitable method for targeting GSCs. For targeting studies, GSCs separated from U87 GBM cells using a magnetic bead separation method were challenged with liposomes encapsulating gemcitabine (GEM) conjugated with a CD133 monoclonal antibody (PEG-lipo-CD133-GEM). An in vitro study showed that conjugation of CD133 antibody significantly enhanced the cytotoxicity of GEM through endocytosis of CD133 surface markers overexpressed on GSCs. The anti-tumor effect of PEG-lipo-CD133-GEM was 15 times higher than that of free GEM, presumably reflecting the specific targeting of the CD133 surface marker by PEG-lipo-CD133-GEM and the enhanced stability and cytotoxicity through the PEGylated liposome formulation in xenograft models. Moreover, monitoring of body weight changes showed that the use of PEGylated liposomes significantly reduced the toxicity of GEM. Taken together, our studies demonstrate that PEG-lipo-CD133-GEM shows promise for the treatment of gliomas in vitro and in xenograft models.


The Canadian contribution to violence risk assessment: history and implications for current psychiatric practice.
Over the past quarter-century, Canadian researchers, clinical practitioners, and policy specialists have made several notable contributions to the broad field of violence risk assessment and management. In part, these contributions have been fostered by major changes in law over this period; in part, they have been spurred by findings from large-scale Canadian prediction--outcome studies. This paper offers references for a range of Canadian-inspired assessment schemes designed to evaluate psychopathy and potential for violence against others.


Unusual presentation of familial Mediterranean fever: role of genetic diagnosis.
To describe the role of molecular analysis in the diagnosis of an unusual presentation of familial Mediterranean fever (FMF). Two patients presenting with prolonged fever without signs and symptoms of serositis are described. FMF was diagnosed by genetic analysis, which disclosed that both patients were homozygous for the M694V mutation of the Mediterranean fever (MEFV) gene. Molecular analysis of FMF should complement the investigation of patients with fever of unknown origin. This test enables a definite diagnosis of the disease and may promote the diagnosis and treatment of patients with an unusual or incomplete clinical picture of FMF.


One Platform Comparison of Polymeric and Lipidic Nanoparticles for the Delivery of Amphotericin B.
Amphotericin B (AmB) is a membrane-acting antibiotic used for the treatment of fungal and protozoal infections. AmB exists in various molecular forms, i.e., monomeric, super-aggregated, and oligomeric forms, where oligomeric forms are highly toxic because of their relative affinity toward cholesterol present over human cell membrane. Hence, the objective of our research work was to study the aggregation state of AmB in two different nanoformulations, i.e., solid lipid nanoparticles (SLNs) and zein-based nanoparticles (PNPs), with the aim of enhancing the fraction of less toxic form of AmB, and a comparative study was performed. The zein and glyceryl monostearate can intercalate the polyenic domain of AmB and thereby hinder the hydrophobic attractions between the AmB molecules, which allows their existence in monomeric forms. The particle size of AmB-SLNs and AmB-PNPs were 378.90 ± 9.50 nm and 184.90 ± 6.00 nm, while zeta potential was -34.97 ± 0.51 mV and +28.93 ± 2.29 mV, respectively. In vitro release studies showed more controlled release of AmB from PNPs (52.48 ± 1.07%) as compared to SLNs (86.33 ± 0.93%). The predominant aggregation state of AmB in both formulations was determined by UV-visible and circular dichroism spectrophotometry, where a higher degree of monomerization of AmB was reported in AmB-SLNs as compared to AmB-PNPs. Toxicity of the nanoformulations was evaluated through hemolysis test, where the results suggested that AmB-SLNs and AmB-PNPs were less hemolytic as compared to pure AmB. The nanoformulations demonstrated the predominant monomeric form of AmB, which may offer higher selectivity index toward microbial membrane.


A WebGIS tool for visualizing and exploring socioeconomic vulnerability to dengue fever in Cali, Colombia.
WebGIS tools have the potential to disseminate the outputs of spatial vulnerability assessments to a wide range of communities, including public health decision-makers. Based on a previous assessment of socioeconomic vulnerability to dengue fever in Cali, Colombia, we developed and used a WebGIS tool to facilitate the visualization, exploration and dissemination of prevailing vulnerabilities to dengue fever in an interactive online environment. Results show that the tool presented here has distinct implications for policy and decision-making as it facilitates spatial prioritisation, both with respect to the intervention areas and the intervention measures needed to reduce human susceptibility and strengthen resilience to the disease.


Role of recanalization in acute stroke outcome: rationale for a CT angiogram-based "benefit of recanalization" model.
In acute middle cerebral artery (MCA) stroke, CT angiographic (CTA) source images (CTA-SI) identify tissue likely to infarct despite early recanalization. This pilot study evaluated the impact of recanalization status on clinical and radiologic predictors of patient outcomes. Of 44 patients undergoing CT/CTA within 6 hours of developing symptoms of proximal MCA ischemia, 19 patients achieved complete proximal MCA (MCA M1) recanalization. Admission National Institutes of Health Stroke Scale (NIHSS) score, onset-to-imaging time, CTA-SI Alberta Stroke Program Early CT Score, MCA M1 occlusion, cerebrovascular collaterals score, and CTA-SI lesion volume were correlated with 3- to 6-month follow-up modified Rankin Scale (mRS). We developed 2 stepwise regression models: one for patients with complete MCA M1 recanalization and one for patients without complete recanalization. Complete and incomplete recanalization groups had similar median admission NIHSS scores (19 versus 19) and mean onset-to-imaging times (2.3 versus 1.9 hours) but different proportions of patients achieving mRS scores 0-2 (74% versus 40%; P = .04). The only independent predictors of clinical outcome in patients with complete recanalization were onset-to-imaging time and admission CTA-SI lesion volume (total model R(2) = 0.75; P = .01). The only independent predictors of outcome in patients with incomplete recanalization were admission CTA-SI lesion volume and NIHSS score (total model R(2) = 0.66; P = .007). Regardless of recanalization status, admission CTA-SI lesion volume was associated with clinical outcome. Recanalization status did, however, affect which variables in addition to CTA-SI volume significantly impacted clinical outcome: time with complete recanalization and NIHSS with incomplete recanalization. This finding may support the development of a model predicting the potential clinical benefit expected with early successful recanalization.


VITAMIN D DEFICIENCY AND PARATOHOMMONIUM INCREASE IN LATE POSTOPERATIVE GASTRIC BYPASS IN ROUX-EN-Y.
Roux-en-Y gastric bypass patients can experience changes in calcium metabolism and hyperparathyroidism secondary to vitamin D deficiency. To evaluate nutritional deficiencies related to the calcium metabolism of patients undergoing gastric bypass with a 10-year follow-up. This is a longitudinal retrospective study of patients submitted to Roux-en-Y gastric bypass at a multidisciplinary clinic located in the Brazilian southeast region. The study investigated the results of the following biochemical tests: serum calcium, ionized calcium, vitamin D, and parathormone (PTH). The generalized estimating equations (GEE) determined the nutritional deficiencies using a significance level of 5%. Among the patients who finished the study (120 months), 82.86% (n=29) had vitamin D deficiency, and 41.94% (n=13) had high PTH. Postoperative time had a significant effect on PTH (p=0.0059). The percentages of patients with vitamin D, serum calcium, and ionized calcium deficiencies did not change significantly over time. One of the outcomes was vitamin D deficiency associated with secondary hyperparathyroidism. These findings reaffirm the importance of monitoring the bone metabolism of patients submitted to Roux-en-Y gastric bypass. Calcium deficiency. Vitamin D deficiency. Secondary hyperparathyroidism.


Cirsiliol induces autophagy and mitochondrial apoptosis through the AKT/FOXO1 axis and influences methotrexate resistance in osteosarcoma.
Osteosarcoma (OS) is the most common primary malignant bone tumor in children and adolescents, with poor outcomes for patients with metastatic disease or chemotherapy resistance. Cirsiliol is a recently found flavonoid with anti-tumor effects in various tumors. However, the effects of cirsiliol in the regulation of aggressive behaviors of OS remain unknown. The effect of cirsiliol on the proliferation of OS cells was detected using a cell counting kit-8 (CCK-8) assay and 5-ethynyl-2'-deoxyuridine (EdU) staining, while cell apoptosis was detected using flow cytometry. Immunofluorescence was applied to visualize the expression level of the mitochondria, lysosomes and microtubule-associated protein light chain 3 (LC3). A computational molecular docking technique was used to predict the interaction between cirsiliol and the AKT protein. The impact of cirsiliol on resistance was investigated by comparing it between a methotrexate (MTX)-sensitive OS cell line, U2OS, and a MTX-resistant OS cell line, U2OS/MTX. Finally, in situ xenogeneic tumor models were used to validate the anti-tumor effect of cirsiliol in OS. Cirsiliol inhibited cell proliferation and induced apoptosis in both U2OS and U2OS/MTX300 OS cells. In addition, treatment with cirsiliol resulted in G2 phase arrest in U2OS/MTX300 and U2OS cells. Cell fluorescence probe staining results showed impaired mitochondria and increased autophagy in OS cells after treatment with cirsiliol. Mechanistically, it was found that cirsiliol targeted AKT by reducing the phosphorylation of AKT, which further activated the transcriptional activity of forkhead Box O transcription factor 1 (FOXO1), ultimately affecting the function of OS cells. Moreover, in situ tumorigenesis experiments showed that cirsiliol inhibited the tumorigenesis and progression of OS in vivo. Cirsiliol inhibits OS cell growth and induces cell apoptosis by reducing AKT phosphorylation and further promotes FOXO1 expression. These phenomena indicate that cirsiliol is a promising treatment option for OS.


Melatonin regulates glucocorticoid receptor: an answer to its antiapoptotic action in thymus.
We have previouslyreported that low doses of melatonin inhibit apoptosis in both dexamethasone-treated cultured thymocytes (standard model for the study of apoptosis) and the intact thymus. Here we elucidate the mechanism by which this agent protects thymocytes from cell death induced by glucocorticoids. Our results demonstrate an effect of melatonin on the mRNA for antioxidant enzymes in thymocytes, also showing an unexpected regulation by dexamethasone of these mRNA. Both an effect of melatonin on the general machinery of apoptosis and a possible regulation of the expression of the cell death related genes bcl-2 and p53 are shown not to be involved. We found melatonin to down-regulate the mRNA for the glucocorticoid receptor in thymocytes (glucocorticoids up-regulate their own receptor). The decrease by melatonin of mRNA levels for this receptor in IM-9 cells (where glucocorticoids down-regulate it) demonstrates that melatonin actually down-regulates glucocorticoid receptor. These findings allow us to propose the effects of melatonin on this receptor as the likely mediator of its thymocyte protection against dexamethasone-induced cell death. This effect of melatonin, given the oxidant properties of glucocorticoids, adds another mechanism to explain its antioxidant effects.


Double inverse-weighted estimation of cumulative treatment effects under nonproportional hazards and dependent censoring.
In medical studies of time-to-event data, nonproportional hazards and dependent censoring are very common issues when estimating the treatment effect. A traditional method for dealing with time-dependent treatment effects is to model the time-dependence parametrically. Limitations of this approach include the difficulty to verify the correctness of the specified functional form and the fact that, in the presence of a treatment effect that varies over time, investigators are usually interested in the cumulative as opposed to instantaneous treatment effect. In many applications, censoring time is not independent of event time. Therefore, we propose methods for estimating the cumulative treatment effect in the presence of nonproportional hazards and dependent censoring. Three measures are proposed, including the ratio of cumulative hazards, relative risk, and difference in restricted mean lifetime. For each measure, we propose a double inverse-weighted estimator, constructed by first using inverse probability of treatment weighting (IPTW) to balance the treatment-specific covariate distributions, then using inverse probability of censoring weighting (IPCW) to overcome the dependent censoring. The proposed estimators are shown to be consistent and asymptotically normal. We study their finite-sample properties through simulation. The proposed methods are used to compare kidney wait-list mortality by race.


Rat Mammary Gland RNA: Incorporation of C14-Formate and Effect of Hormones and 7,12-Dimethylbenz[alanthracene.
In female rats, the incorporation of sodium C(14)-formate into mammary gland RNA decreases immediately after a singlefeeding of 20 milligrams of 7,12-dimethylbenz[a] anthracene. In males, there is a gradual increase in the incorporation. In castrated rats, the decrease or increase of C(14)-formate incorporation is dependent on the presence of estrogen or androgen, respectively.


Delineating the regions of human transferrin involved in interactions with transferrin binding protein B from Neisseria meningitidis.
Pathogenic bacteria in the Neisseriaceae possess a surface receptor mediating iron acquisition from human transferrin (hTf) that consists of a transmembrane iron transporter (TbpA) and a surface-exposed lipoprotein (TbpB). In this study, we used hydrogen/deuterium exchange coupled to mass spectrometry (H/DX-MS) to elucidate the effects on hTf by interaction with TbpB or derivatives of TbpB. An overall conserved interaction was observed between hTf and full-length or N-lobe TbpB from Neisseria meningitidis strains B16B6 or M982 that represent two distinct subtypes of TbpB. Changes were observed exclusively in the C-lobe of hTf and were caused by the interaction with the N-lobe of TbpB. Regions localized to the 'lip' of the C1 and C2 domains that flank the interdomain cleft represent sites of direct contact with TbpB whereas the peptides within the interdomain cleft that encompass iron binding ligands are inaccessible in the closed (holo) conformation. Although substantial domain separation upon binding TbpB cannot be excluded by the H/DX-MS data, the preferred model of interaction involves binding hTf C-lobe in the closed conformation. Alternate explanations are provided for the substantial protection from deuteration of the peptides encompassing iron binding ligands within the interdomain cleft but cannot be differentiated by the H/DX-MS data.


Protective effect of a polyphenols-rich extract from Inonotus Sanghuang on bleomycin-induced acute lung injury in mice.
Mushroom Phellinus linteus ("Sanghuang" in Chinese) is a popular medicinal polypore used to treat several disorders through its various biological functions. Inonotus sanghuang is claimed to produce general immune-potentiating and strengthening, anti-inflammatory, anti-tumor and anti-microbial properties, but its effect on acute lung inflammation and oxidative stress are not clearly understood. To determine the effect and mechanism of the polyphenols-rich ethyl acetate fraction from wild I. sanghuang extract (ISE) on acute lung injury (ALI) induced by bleomycin (BLM), female C57BL/6 mice were fed ISE (0%, 0.15% or 0.6% in diet) for 4 weeks prior to challenge with BLM. Bronchoalveolar lavage fluid (BALF) from lung, spleen and lung tissues were collected on day 3 after BLM challenge for histological, oxidative stress, molecular and biochemical analysis. ISE supplementation improved pathological features in lung injury scores and reduced lung wet-to-dry ratios. Moreover, ISE reduced inflammatory cell infiltration and the pro-inflammatory cytokines including IL-1β, IL-6 and TNF-α in BALF, decreased the MPO activity and the MDA level and increased the SOD, CAT and GSH-Px activities in lung tissue homogenates. Further mechanism analysis demonstrated that dietary ISE inhibited NF-κB signal. Finally, peripheral immune function analysis showed that ISE had less effect on immune response including splenocyte producing inflammatory cytokines and T cell proliferation except for IL-1β and IL-2. Our findings indicate the possibility that dietary ISE attenuates ALI induced by BLM through correcting the inflammation and oxidation balance at least in part via inhibiting NF-κB signal in vivo, suggesting that ISE might be a valuable medicinal food effective in improving lung injury.


Dentofacial growth and development in Homo sapiens: evidence from perinatal individuals from Punic Carthage.
Study of perinatal individuals from 7-4th c. B.C. Punic Carthage and specimens from more recent sources elucidates: 1. variability in the formation of the infraorbital foramen; 2. the nature of, and variability in, the expression of the incisive suture. With regard to the latter, and in conjunction with data on the embryological formation of the upper jaw and dentition, the following generalization appears warranted in Homo sapiens. The incisors and canine may come to erupt in the alveolar bone associated with the premaxillary region. Combination of these data with those on the timing of tooth germ development and their forward migration yields an explanation of the dental anomalies associated with anterior facial cleft and a prediction of when this disorder occurs.


[Analysis of the actual state of nutrition of elderly patients fed via PEG tube].
Over a period of six months starting from October 1, 2001, within the frameworks of quality checks in nursing homes and the examination to check the need of care, the data of 290 persons supplied with a PEG-tube were collected. In only 60% of all cases the individual weight had been recorded. For 40% of the group the most important regulatory parameter for calorie intake was not available. Weights which had not been measured were estimated by the MDK experts. In total, 27% of the PEG patients were underweight. Assuming an activity factor of 1.2 the minimal required amount of calories was calculated on the basis of age, height, weight and sex and compared with the actual calorie intake. Up to 19% of the patients with PEG were mainly fed orally. About half of these people could probably be nourished without a PEG. 66% of all people were fed exclusively via PEG. 47% of them received less calories than their basal metabolic rate (BMR), 70% received less than their minimal energy requirement. On average, the calorie intake was 171 kcal below the BMR. In those cases, where either an external nutrition expert was consulted or the actual weight was known, the share of patients receiving less than the required amount of calories was slightly lower. Within the framework of persons surveyed for the check of a need of care, the situation of those being taken care of in their private home surroundings was considerably better than of those who were living in a nursing home.


Spinal cord stimulation facilitates functional walking in a chronic, incomplete spinal cord injured.
This paper describes a treatment paradigm to facilitate functional gait in a quadriplegic, ASIA C spinal cord injured (SCI), wheelchair-dependent subject who presented with some large fiber sensation, sub-functional motor strength in all lower limb muscles, and moderate spasticity. The study utilizes partial weight bearing therapy (PWBT) followed by epidural spinal cord stimulation (ESCS) with the assumption that both treatments would be necessary to elicit a well organized, near effortless functional gait with a walker. Function is defined in terms of accomplishing task-specific activities in the home and community. To demonstrate the feasibility and benefits of combined PWBT and ESCS therapies aimed at promoting functional gait in a wheelchair-dependent ASIA C SCI subject. The Clinical Neurobiology and Bioengineering Research Laboratories at Good Samaritan Regional Medical Center, Phoenix, Arizona, USA, and the Department of Bioengineering, Arizona State University, Tempe, Arizona, USA. The study began with the application of PWBT. The subject walked on the treadmill until a plateau in gait rhythm generation was reached. Subsequently, ESCS, applied to the lumbar enlargement, was utilized to facilitate PWBT and, later, over-ground walking for a standard distance of 15 m. Gait performance was analyzed by measuring average speed, stepping symmetry, sense of effort, physical work capacity, and whole body metabolic activity. PWBT led to improved stereotypic stepping patterns associated with markedly reduced spasticity, but was insufficient for over-ground walking in terms of safety, energy cost, and fatigue. ESCS with PWBT generated immediate improvement in the subject's gait rhythm when appropriate stimulation parameters were used. When compared to the non-stimulated condition, over-ground walking with ESCS across a 15 m distance was featured by a reduction in time and energy cost of walking, sense of effort, and a feeling of 'lightness' in the legs. After a few months of training, performance in speed, endurance, and metabolic responses gradually converged with/without ESCS at this short distance, suggesting a learned response to these conditions. However, at longer distances (eg, 50-250 m), performance with ESCS was considerably superior. The subject was able to perform multiple functional tasks within the home and community with ESCS. We propose that ESCS augments the use-dependent plasticity created by PWBT and may be a valuable adjunct to post-SCI treadmill training in ASIA C subjects. We also conclude that ESCS elicits greater activation of an oxidative motor unit pool, thereby reducing the subject's sense of effort and energetic cost of walking.


[Nitrogen metabolism and its control mechanisms].
N intake in the form of protein has neither got an upper nor a lower limit for agricultural working animals within a diet and there is no control mechanism for it. A high surplus of certain amino acids results in a reduction of feed intake. N excretion in faeces depends on 1) the excretion of N containing indigestible feedstuffs, 2) bacterial nitrogen synthesis in the large intestine and 3) the excretion of true endogenous N containing substances (digestion enzymes, intestinal epithelium, N containing endogenous secretion). There are no other control mechanisms for N excretion in faeces. N excretion in urine mainly comprises the nitrogen from the degeneration of amino acids and nucleic acids. The interrelations between urea, NH3, allantoin, creatine and creatinine, uric acid and hippuric acid depend on the species (monogastric or ruminants), on the nitrogen and N amount consumed and on the recycling ratio of the amino acids. The absolute amount of N excretion is not subject to any control mechanism, it depends on the intake of protein and NPN substances, the interim stages, however, which lead to the formation of excretory products, are intermediately controlled. The most important interim stage is protein biosynthesis, which is a fixed, intermediately controlled value in maintenance level. Under growth conditions only, the protein synthesis quota can exceed the protein degradation quota of the total organism (positive N balance). The control mechanisms of protein biosynthesis have, according to current knowledge, the following structure: Stimulation: 1) growth hormone (STH) stimulates protein synthesis by means of somatomedins; 2) hormones of the thyroid gland (T4 and T3) are controlled by the hormone stimulating the thyroid gland (TSH); 3) insulin. Inhibition: 1) somatostatin inhibits STH, TSH and insulin; 2) cortisol directly inhibits protein synthesis and stimulates protein degradation. The control mechanisms of protein turnover in addition to genetic coding and proteolysis extend in the framework of evolution over the period of 3,400 million years from the existence of the bacterial cell to the development of mammals, which is 74% of the age of the earth and approximately 90% since the existence of the first traces of life. The control mechanisms of protein turnover in mammals do not permit gene manipulation in protein synthesis as in bacterial cells since the control mechanisms mentioned are missing there.


Characterization and functional expression of a ubiquitously expressed tomato pectin methylesterase.
Pectin methylesterase (PME), a ubiquitous enzyme in plants, de-esterifies the methoxylated pectin in the plant cell wall. We have characterized a PME gene (designated as pmeu1) from tomato (Lycopersicon esculentum) with an expression that is higher in younger root, leaf, and fruit tissues than in older tissues. Hypocotyls and epicotyls show higher accumulation of pmeu1 transcripts compared with cotyledons. pmeu1 represents a single-copy gene in the tomato genome. Comparison of the deduced amino acid sequence of pmeu1 with other PME homologs showed that the N-terminal halves are highly variable, and the C-terminal halves are relatively conserved in plant PMEs. Constitutive expression of a fruit-specific PME antisense gene does not affect the level of pmeu1 transcripts in vegetative tissues but does lower the level of PMEU1 mRNA in developing tomato fruits. These results suggest that there exists developmentally regulated silencing of pmeu1 by a heterologous PME antisense gene. Expression of pmeu1 in tobacco (Nicotiana tabacum) under the control of the cauliflower mosaic virus 35S promoter caused up to a 4-fold increase in PME specific activity that was correlated with the accumulation of PMEU1 mRNA. In vitro transcription-translation analyses show that pmeu1 encodes a 64-kD polypeptide, whereas transgenic tobacco plants expressing pmeu1 accumulate a new 37-kD polypeptide, suggesting extensive posttranslational processing of PMEU1. These results are the first evidence, to our knowledge, of the functional characterization of a PME gene and the extensive modification of the encoded polypeptide.


Evaluation of fetal liver volume by tridimensional ultrasound in women with gestational diabetes mellitus.
To evaluate the effect of gestational diabetes mellitus (GDM) on fetal liver growth during the third trimester. We performed a longitudinal study of pregnant women recruited at the time of GDM screening (24 to 28 weeks of gestation), with follow-up visits at 32 weeks, 36 weeks, and delivery. Women with GDM were followed with nutritional recommendations and insulin when necessary according to the Canadian Diabetes Association guidelines. Fetal liver volume was evaluated using 3-D ultrasound at each antenatal visit, and fetal liver growth was compared between women with and without GDM. Twenty-seven women were recruited, 10 with normal glucose tolerance (NGT) and 17 with confirmed GDM, five who required insulin and 12 who were treated by diet only. We found no difference in fetal liver volume between groups at any of the three visits, and median birth weight was also similar between groups. On the other hand, we found a strong correlation between fetal liver volume at 36 weeks' gestation and birth weight (ρ = 0.61, P < 0.001). In our preliminary study, we found that fetal liver volume could be a strong predictor of infant birth weight independent of GDM status. This suggests that fetal liver volume of offspring of women with NGT is similar to that of offspring of women with GDM treated following recommended targets. Larger studies are required.


Determinants of early-stage hearing loss among a cohort of young workers with 16-year follow-up.
The authors had a unique opportunity to study the early impacts of occupational and recreational exposures on the development of noise-induced hearing loss (NIHL) in a cohort of 392 young workers. The objectives of this study were to estimate strength of associations between occupational and recreational exposures and occurrence of early-stage NIHL and to determine the extent to which relationships between specific noise exposures and early-stage NIHL were mitigated through the use of hearing protection. Participants were young adults who agreed to participate in a follow-up of a randomised controlled trial. While the follow-up study was designed to observe long-term effects (up to 16 years) of a hearing conservation intervention for high school students, it also provided opportunity to study the potential aetiology of NIHL in this worker cohort. Study data were collected via exposure history questionnaires and clinical audiometric examinations. Over the 16-year study period, the authors documented changes to hearing acuity that exceeded 15 dB at high frequencies in 42.8% of men and 27.7% of women. Analyses of risk factors for NIHL were limited to men, who comprised 68% of the cohort, and showed that risks increased in association with higher levels of the most common recreational and occupational noise sources, as well as chemical exposures with ototoxic potential. Use of hearing protection and other safety measures, although not universal and sometimes modest, appeared to offer some protection. Early-stage NIHL can be detected in young workers by measuring high-frequency changes in hearing acuity. Hearing conservation programmes should focus on a broader range of exposures, whether in occupational or non-occupational settings. Priority exposures include gunshots, chainsaws, power tools, smoking and potentially some chemical exposures.


AIDS: an update.
Although there is no epidemiologic evidence for increased spread of the disease to non-high-risk groups, many investigators speculate that AIDS may be caused by an infectious agent that can be transmitted through blood or blood products. If this hypothesis proves to be correct, dramatic public measures will have to be implemented with respect to blood banking policies.


Improved Biocompatibility of Amino-Functionalized Graphene Oxide in Caenorhabditis elegans.
Graphene oxide (GO) holds high promise for diagnostic and therapeutic applications in nanomedicine but reportedly displays immunotoxicity, underlining the need for developing functionalized GO with improved biocompatibility. This study describes adverse effects of GO and amino-functionalized GO (GONH2 ) during Caenorhabditis elegans development and ageing upon acute or chronic exposure. Chronic GO treatment throughout the C. elegans development causes decreased fecundity and a reduction of animal size, while acute treatment does not lead to any measurable physiological decline. However, RNA-Sequencing data reveal that acute GO exposure induces innate immune gene expression. The p38 MAP kinase, PMK-1, which is a well-established master regulator of innate immunity, protects C. elegans from chronic GO toxicity, as pmk-1 mutants show reduced tissue-functionality and facultative vivipary. In a direct comparison, GONH2 exposure does not cause detrimental effects in the wild type or in pmk-1 mutants, and the innate immune response is considerably less pronounced. This work establishes enhanced biocompatibility of amino-functionalized GO in a whole-organism, emphasizing its potential as a biomedical nanomaterial.