Two-phase anaerobic digestion of lignocellulosic hydrolysate: Focusing on the acidification with different inoculum to substrate ratios and inoculum sources.
Biogas production from lignocellulosic hydrolysate is of great potential for lignocellulosic materials. Two-phase anaerobic digestion was proposed in this study. Acidogenic fermentation was carried out with corn straw hydrolysate as feedstock for volatile fatty acids (VFAs) production. Using anaerobic sludge (AnS), different inoculum to substrate ratios (ISRs) of 0.5:1, 1:1 and 2:1 were investigated. The highest VFAs yield was obtained at ISR of 0.5:1.VFAs composition analysis showed that butyric acid was the predominant acid, followed by acetic acid and propionic acid. The effects of AnS and aerobic sludge (AeS) on the acidogenic performance of hydrolysate were compared. The optimum VFA yields were 0.38 g/g COD-added for AnS and 0.32 g/g COD-added for AeS with HRT of 5 d, respectively. The bacterial diversities of inocula and digestates were analyzed by high-throughput sequencing. Two origins of inocula had distinct bacterial structures, but they did share core communities that included Firmicutes, Chloroflexi, Proteobacteria and Bacteroidetes at phylum level. The bacterial communities of both digestates changed significantly as compared with those in inoculum. Firmicutes was absolutely dominant in all the bacterial species. Therefore, the AeS could be an option as the acidogenic inoculum. The microbial information will be beneficial for the enrichment and acclimatization of microbes. In methanogenic process, VFAs obtained in acidogenic stage could be efficiently converted into methane. The ultimate methane yield at organic loading rate (OLR) of 8 g/L·d could reach 290 mL CH4/g COD-added and 279 mL CH4/g COD-added for AnS and AeS acidified digestate. Two-phase anaerobic digestion was proved to be suitable for bioconversion of lignocellulosic hydrolysate into biogas.


Gene expression and activity of carbonic anhydrase in salinity stressed Penaeus monodon.
Carbonic anhydrase (CA) was identified by differential display PCR analysis as one of the differentially expressed genes in the gills of low salinity stressed (transferred from 25 to 3 ppt) Penaeusmonodon. To further characterize the role of CA in the regulation of salinity stress, the cDNA sequence of P.monodon carbonic anhydrase (PmCA) was attained by rapid amplification of cDNA ends and found to have a total length of 1194 bp. The deduced amino acid of PmCA shares 73% sequence identity with the CA homologue recently isolated from the crab, Callinectessapidus. Real time RT-PCR and enzymatic activity analyses were employed to determine the changes in the PmCA mRNA expression and total CA activity, respectively, after shrimps were transferred from 25 to 3 ppt salinities for up to 2 weeks. Compared to the CA level in the control group (25 ppt), PmCA mRNA was significantly increased in shrimp gills at 24 h after hypo-osmotic stress. In contrast, the epipodites and antennal gland displayed decreased levels of mRNA expression. The gross CA enzymatic activity after hypo-osmotic stress was increased in the shrimp gills but remained stable in the epipodites and antennal gland.


A review of composition studies of Cameroon traditional dishes: macronutrients and minerals.
This paper reviews published data that contributes to the knowledge of the ingredients and nutrients of Cameroon traditional dishes. Macronutrient (energy, carbohydrates, protein, total fat, fibre and ash) and mineral (iron, zinc, magnesium, calcium, phosphorus, copper, manganese, potassium, sodium and selenium) data are presented for 117 commonly consumed dishes from three eco-regions. Tables providing an overview of the main ingredients and nutrient values (range of means per 100g edible portion) are presented. Considerable variability in nutrient values has been reported among dishes. Water contents range from 29.8 to 95.9g/100g edible portion while energy values range from 12 to 403kcal/100g. Energy yielding-constituents are the major nutrients recorded in published data, followed by iron, zinc and magnesium.


Locating the evidence base for musculoskeletal nursing: an overview of the rheumatology nursing literature.
The main aim of this study was to locate and classify publications, particularly research publications, produced by nurses working and/or researching in the area of rheumatology in order to develop a research strategy for a newly established academic and clinical nursing unit. Database searches were undertaken to identify journal articles. Theses, dissertations, research reports, conference abstracts and books were also included in the study. The publications once located were entered into a Reference Manager electronic database and the facilities of this package were used to analyse and classify the data. The findings of the study demonstrated that over the past 40 years nurses have been steadily increasing the publication output in this area and in the last 10 years there has been a substantial increase in all publications, but particularly research and investigative articles. However there is a lack of clinical research in this area and this needs to be addressed in future as nurses develop more advanced roles in this speciality. The main limitations was the difficulty in accurately identifying nursing publications because of inadequate details of authorship.


New mescaline-related N-acylhydrazone and its unsubstituted benzoyl derivative: Promising metallophores for copper-associated deleterious effects relief in Alzheimer's disease.
Alzheimer's disease (AD) is related to the presence of extracellular aggregated amyloid-β peptide (Aβ), which binds copper(II) with high affinity in its N-terminal region. In this sense, two new 1-methylimidazole-containing N-acylhydrazonic metallophores, namely, X1TMP and X1Benz, were synthesized as hydrochlorides and characterized. The compound X1TMP contains the 3,4,5-trimethoxybenzoyl moiety present in the structure of mescaline, a natural hallucinogenic protoalkaloid that occurs in some species of cacti. Single crystals of X1Benz, the unsubstituted derivative of X1TMP, were obtained. The experimental partition coefficients of both compounds were determined, as well as their apparent affinity for Cu2+ in aqueous solution. Ascorbate consumption assays showed that these N-acylhydrazones are able to lessen the production of ROS by the Cu(Aβ)-system, and a short-time scale aggregation study, measured through turbidity and confirmed by TEM images, revealed their capacity in preventing Aβ fibrillation at equimolar conditions in the presence and absence of copper. 1H15N HSQC NMR experiments demonstrated a direct interaction between Aβ and X1Benz, the most soluble of the compounds. The Cu2+ sequestering potential of this hydrazone towards Aβ was explored by 1H NMR. Although increasing amounts of X1Benz were unexpectedly not efficient at removing the metal-induced perturbations in Aβ backbone amides, the broadening effects observed on the compound's signals indicate the formation of a ternary Aβ‑copper-X1Benz species, which can be responsible for the observed ROS-lessening and aggregation-preventing activities. Overall, the N-acylhydrazones X1TMP and X1Benz have shown promising prospects as agents for the treatment of AD.


Targeting corticostriatal transmission for the treatment of cannabinoid use disorder.
It is generally assumed that the rewarding effects of cannabinoids are mediated by cannabinoid CB1 receptors (CB1Rs) the activation of which disinhibits dopaminergic neurons in the ventral tegmental area (VTA). However, this mechanism cannot fully explain novel results indicating that dopaminergic neurons also mediate the aversive effects of cannabinoids in rodents, and previous results showing that preferentially presynaptic adenosine A2A receptor (A2AR) antagonists counteract self-administration of Δ-9-tetrahydrocannabinol (THC) in nonhuman primates (NHPs). Based on recent experiments in rodents and imaging studies in humans, we propose that the activation of frontal corticostriatal glutamatergic transmission constitutes an additional and necessary mechanism. Here, we review evidence supporting the involvement of cortical astrocytic CB1Rs in the activation of corticostriatal neurons and that A2AR receptor heteromers localized in striatal glutamatergic terminals mediate the counteracting effects of the presynaptic A2AR antagonists, constituting potential targets for the treatment of cannabinoid use disorder (CUD).


Sero-epidemiology of Bordetella pertussis in England and Wales.
The incidence of pertussis infection can be estimated in the population by defining a single high titre of anti pertussis toxin (PT) immunoglobulin G (IgG) antibody predictive of recent infection. Sera samples collected in 1986, 1996 or annually between 1987 and 1998 were tested for anti-PT IgG antibody. In 1996, the age-adjusted prevalence of pertussis infection was 1.2% and was higher in children than in adults. Amongst samples collected annually, older age and female sex, but none of the temporal variables, were associated with a serologically defined pertussis infection. There is an important incidence of infection in the population, which is greater amongst children than adults, but there is only limited evidence of a correlation with epidemic cycles.


Bacterial expression systems for recombinant protein production: E. coli and beyond.
Escherichia coli expression system continues to dominate the bacterial expression systems and remain to be the preferred system for laboratory investigations and initial development in commercial activities or as a useful benchmark for comparison among various expression platforms. Some new developments in overcoming its shortcomings are reviewed in this paper, including antibiotics-free selection plasmids, extracellular production, and posttranslational modifications. The ability for E. coli to make mg glycosylated proteins promises even broader applications of the E. coli system in the future. Significant progresses have also been made over the past few years in alternative bacterial expression systems. Notably, the Lactoccocus lactis system has proven to be a viable choice for membrane proteins. Additionally, several Pseudomonas systems were developed and achieved product titers comparable to E. coli systems. Other bacterial systems such as Streptomyces, coryneform bacteria, and halophilic bacteria offer advantages in some niche areas, providing more choices of bacterial expression systems for recalcitrant proteins.


Endotoxin protection against pulmonary oxygen toxicity and its reversal by acetyl salicylic acid: role of eicosanoid production by broncho-alveolar lavage cells.
Small doses of endotoxin markedly increase the survival rate of adult rats exposed to 98% oxygen for periods that are normally lethal. The lysine salt of acetyl salicylic acid (L-ASA) partially reverses this protective effect of endotoxin. In this pilot study we investigated the level of eicosanoid production by broncho-alveolar lavage (BAL) cells and found that BAL cells of endotoxin protected rats, present in abundance, have an equal or increased capacity of HHT, 15-HETE, 12-HETE, LTB4 and 5-HETE production. These data suggest that production of the lipoxygenase products by BAL cells does not seem to play an important role in the pathogenesis of pulmonary oxygen toxicity. We did not find any indication for the occurrence of shunting of arachidonic acid metabolism to the lipoxygenase pathway as an explanation for the reversal of endotoxin's protective action by L-ASA.


Work on the cutting edge: metallographic investigation of Late Bronze Age tools in southeastern Lower Austria.
This paper analyses 20 Late Bronze Age (ca 1080-800 BC) copper alloy objects to discern their manufacture and the skills of local craftsmen. Several tools and jewellery were studied that originated from a bronze workshop located immediately next to the Prigglitz-Gasteil copper ore mining site and several contemporaneous sites in the surrounding area. The samples were studied with optical microscopy (microstructurally), and SEM-EDXS and XRF (chemical analyses). Our analyses are part of a larger study and suggest that the Prigglitz region's bronze production was not standardized. Particular alloys do not seem to have been chosen for object types or due to their intended use-function. Notably, approximately 20% of the objects contain unalloyed copper inclusions, which are most likely a result of the incomplete mixing of scrap metals and alloys during their production.


Role of social capital in disaster risk management: a theoretical perspective in special reference to Odisha, India.
Social capital being the network of relationship plays a significant role in managing risk and crisis situation saving human lives from an adverse effect and impact of disasters. The authors and scholars across many countries through their disaster studies show that social capital with the form of bonding, bridging and linking rescued the victims of disasters, evacuated the most vulnerable people to safer places, provided food and drinking water and other basic amenities to the victims of disasters, provided homes to the homeless and solaced to those who were psychologically depressed. However, the literature review further specified that while bonding and bridging social capital extended support to the victims of disasters in abundance, the linking social capital on the other hand supported only to those who are in close contact with the persons and institutions in power and authority. Review of literature is done purely through qualitative research methodology which has had read 86 relevant articles published within the last 10- to 12-year period of time. This review paper both contributes to the social capital literature and helps those responsible for planning and policy framework to effectively manage future disasters and safeguard lives and properties and enhance capacities of the most vulnerable people to absorb shocks and stress successfully.


Undetectable phospho-STAT1 in peripheral blood mononuclear cells from patients with chronic hepatitis C who do not respond to interferon-alpha therapy.
Recent studies have suggested that phosphorylated signal transducers and activators of transcription 1 (STAT1) plays an important role in interferon (IFN)-mediated biological functions, including antiviral activity. Moreover, it has been demonstrated that suppressors of the cytokine signal 1 (SOCS1) negatively regulates IFN activities. To investigate the involvement of phospho-STAT1 in the response to IFN-alpha therapy in patients with chronic hepatitis C and to evaluate the negative regulatory effect of SOCS1 on STAT1 activation. Sixty-five patients with chronic hepatitis C and 25 healthy subjects were enrolled. Twenty-five of the patients had never been treated with IFN-alpha therapy (naive), while the remaining 40 patients had. The IFN-treated patients were divided into sustained responders (SRs) or non-responders (NRs) on the basis of their response to the antiviral therapy. Peripheral blood mononuclear cells (PBMCs) were obtained from each patient and control, and were either stimulated with IFN-alpha or left unstimulated. Total STAT1, phospho-STAT1 and SOCS1 were revealed by means of Western blot. Total STAT1 was equally expressed in unstimulated and stimulated PBMCs from all patients and controls. One hundred percent of the stimulated PBMCs from healthy controls and SRs, 96% from naive subjects, and 30% from NRs showed detectable phospho-STAT1. By contrast, 70% of the stimulated PBMCs from NRs showed undetectable phospho-STAT1. We have demonstrated that phospho-STAT1 proteins in 70% of patients with chronic hepatitis C who do not respond to IFN treatment are undetectable, which suggests that this protein may be involved in the mediation of IFN sensitivity. The down-regulation of the Jak-STAT pathway because of SOCS1 expression may be one of the possible underlying mechanisms involved in resistance to IFN.


Ex Vivo Radiation Leads to Opposing Neurite Growth in Whole Ganglia vs Dissociated Cultured Pelvic Neurons.
Prostatic radiation therapy (RT) often causes erectile dysfunction (ED) and the mechanisms governing RT-induced ED are unclear with a lack of therapeutic strategies. To determine the effects of ex vivo RT on major pelvic ganglion (MPG) neuron survival, and neurite growth in whole vs dissociated culture. MPGs were removed and irradiated (0 or 8 Gy) from male Sprague Dawley rats. For dissociated culture, MPG neurons were digested in collagenase/dispase and cultured on coverslips. Immunofluorescent staining for beta-tubulin III (TUBB3; neuron marker), neuronal nitric oxide synthase (nNOS; nitrergic marker), tyrosine hydroxylase (TH; sympathetic marker), and terminal deoxynucleotidyl transferase dUTP nick end labeling assessed neurite length, branching, autonomic neuron density, and apoptosis. For whole organ culture, MPGs were grown in Matrigel. Gene expression of apoptotic markers (caspase 1, 3), TUBB3, nNOS, TH, and Schwann cells (Sox10, Krox20, glial fibrillary acid protein) was measured in whole organ cultured MPGs by quantitative polymerase chain reaction. After 72 hours, neurite length, branching, autonomic neuron density, and apoptosis were assessed, and gene expression was measured. RT increased apoptosis in dissociated neurons measured by terminal deoxynucleotidyl transferase dUTP nick end labeling (P < .001) and whole MPG culture via upregulation of caspase 3 gene expression (P < .05). Nitrergic neurons were markedly decreased in irradiated dissociated culture (P < .05), while nNOS gene expression was upregulated in irradiated whole organ culture (P < .05). The proportion of dissociated sympathetic neurons and whole organ TH gene expression remained unchanged after RT. Interestingly, RT dissociated neurites were 22% shorter than controls, while RT whole organ neurites were 15% longer than controls (P < .01). MPG Schwann cells markers (Sox10, Krox20) were elevated after RT in whole organ culture. Prostatic RT leads to increased neuronal cell death and less erectogenic nitrergic neurons contributing to ED. The advantages of dissociated neuron culture include distinct neurites which are easily measured for apoptosis, length/branching, and specific neuron types. In contrast, whole MPG culture is advantageous as it contains all the supporting cells present in vivo. The 2 different culture methods demonstrated opposing neurite growth after RT indicating the importance of supporting cell network to promote pelvic neuron neuritogenesis and survival following RT. Randolph JT, Pak ES, Koontz BF, et al. Ex Vivo Radiation Leads to Opposing Neurite Growth in Whole Ganglia vs Dissociated Cultured Pelvic Neurons. J Sex Med 2020;17:1423-1433.


Family's socioeconomic profile at birth and offspring mortality until midlife - The Northern Finland Birth Cohort 1966 study.
Family's socioeconomic profile collected prenatally is known to predict offspring mortality during early life, but it remains unclear whether it has the potential to predict offspring mortality until later life. In this study, 12,063 individuals belonging to the Northern Finland Birth Cohort 1966 were followed up from mid-pregnancy for 52 years (570,000 person years). Five distinct socioeconomic profiles were identified by latent class analysis based on mother's marital status, education, and occupation; father's occupation; number of family members; location of residence, room count, and utilities; and family's wealth. The classes were highest status families (15.4% of the population), small families (22.1%), larger families (15.4%), average wealth families (23.4%), and rural families (23.3%). Their associations to offspring mortality, via linkage to national offspring death records, were analysed by Cox regression, stratified by sex and age groups (0-19, 20-38 and 40-52 years). In total, mortality was 9.2% among male and 5.0% among female offspring. Risk for midlife mortality was higher among male offspring from larger families (hazard ratio 2.19, 95% confidence interval 1.32-3.63), average wealth families (1.66, 1.02-2.73) and rural families (1.63, 1.00-2.68), relative to offspring from highest status families. It seems that family's socioeconomic profile constructed prenatally has predictive value for midlife mortality among male offspring. Premature mortality of men and women seem to be two distinct phenomena with differing underlying factors as socioeconomic profile was not associated with mortality among female offspring.


Ultrasound evaluation of diaphragm function in patients with cervical spinal cord injury: case report.
Ultrasound evaluation of diaphragm function allows the detection of diaphragm dysfunction and the adaptation of ventilatory support in patients admitted to intensive care units. The studied patient had a C3 spinal cord injury. Ultrasound evaluation of diaphragm mobility showed that the patient suffered diaphragm dysfunction. A tracheotomy was indicated, and early ventilatory support was initiated. Ultrasound evaluation of diaphragm function in patients with cervical spinal cord injury is a useful and simple technique. It provides fast and reliable data for the diagnosis of respiratory insufficiency of neuromuscular origin.


The past, present, and future physiology and pharmacology of glucagon.
The evolution of glucagon has seen the transition from an impurity in the preparation of insulin to the development of glucagon receptor agonists for use in type 1 diabetes. In type 2 diabetes, glucagon receptor antagonists have been explored to reduce glycemia thought to be induced by hyperglucagonemia. However, the catabolic actions of glucagon are currently being leveraged to target the rise in obesity that paralleled that of diabetes, bringing the pharmacology of glucagon full circle. During this evolution, the physiological importance of glucagon advanced beyond the control of hepatic glucose production, incorporating critical roles for glucagon to regulate both lipid and amino acid metabolism. Thus, it is unsurprising that the study of glucagon has left several paradoxes that make it difficult to distill this hormone down to a simplified action. Here, we describe the history of glucagon from the past to the present and suggest some direction to the future of this field.


Mortality and morbidity of surgical and transcatheter mitral valve repair in octogenarians: A systematic review.
Valvular heart disease (VHD) is a prominent problem in healthcare today with mitral regurgitation (MR) being the leading cause of VHD in the elderly population. While mitral valve repair (MVr) surgery is one of the only options for the end-stage disease, octogenarians are often denied MVr due to concerns with operative mortality and postoperative morbidity. To provide information on this underrepresented group of surgical patients, a systematic approach was taken to review the mortality and morbidity rates of octogenarians who received MVr. Pubmed and Medline were searched for articles containing outcomes of octogenarians receiving surgical mitral valve repair (SMVr) or transcatheter mitral valve repair (TMVr) published after 2000. Ten articles met the inclusion criteria for a total of 7968 patients included in the analysis using Microsoft Excel, Version 2105. Short-term mortality rates for SMVr and TMVr were 2.6% and 1.4% for in-hospital, and 7.8% and 3.3% for 30 days, respectively. The average incidence of stroke, acute kidney injury, infection, and major bleeding for SMVr were 3.2%, 11.2%, 7.7%, and 24%, respectively, and 0.3%, 6.7%, 2.7%, and 7.9% for TMVr, respectively. Octogenarians receiving SMVr or TMVr experienced similar rates of short-term mortality and morbidity as younger populations, and when considering life expectancy, midterm mortality was also similar. With these results, denying octogenarians MVr operations based on age alone should be reconsidered. Depending on risk factors and comorbidities, either SMVr or TMVr is a viable and relatively safe option for octogenarians with severe MR.


Effects of dietary corticosterone on the growth and metabolism of immature Gallus domesticus.
Four-week-old chicks were fed on diets containing corticosterone (0, 5, 10, 20, and 40 mg/kg) for 9 days. Food intake, weight gain, plasma concentrations of corticosterone and other constituents, some organ weights, and adrenal cholesterol concentration were measured. Plasma corticosterone concentration was increased (P less than 0.05) by dietary corticosterone, particularly at the two highest rates of inclusion. The concentrations of glucose, cholesterol, triglyceride, protein, and uric acid in the plasma increased (P less than 0.05) with increasing concentrations of dietary corticosterone. Growth rate and relative weight of the cloacal bursa declined (P less than 0.05), while the relative weight of the liver increased (P less than 0.01), with increasing dietary corticosterone; relative weights of the spleen and adrenals were not significantly affected, nor was the adrenal concentration of cholesterol. Abdominal fat was increased sixfold by the two highest corticosterone concentrations.


Estradiol plays a role in regulating the expression of lysyl oxidase family genes in mouse urogenital tissues and human Ishikawa cells.
The lysyl oxidase (LOX) family encodes the copper-dependent amine oxidases that play a key role in determining the tensile strength and structural integrity of connective tissues by catalyzing the crosslinking of elastin or collagen. Estrogen may upregulate the expression of LOX and lysyl oxidase-like 1 (LOXL1) in the vagina. The objective of this study was to determine the effect of estrogen on the expression of all LOX family genes in the urogenital tissues of accelerated ovarian aging mice and human Ishikawa cells. Mice and Ishikawa cells treated with estradiol (E2) showed increased expression of LOX family genes and transforming growth factor β1 (TGF-β1). Ishikawa cells treated with TGF-β1 also showed increased expression of LOX family genes. The Ishikawa cells were then treated with either E2 plus the TGF-β receptor (TGFBR) inhibitor SB431542 or E2 alone. The expression of LOX family genes induced by E2 was reduced in the Ishikawa cells treated with TGFBR inhibitor. Our results showed that E2 increased the expression of the LOX family genes, and suggest that this induction may be mediated by the TGF-β signal pathway. E2 may play a role in regulating the expression of LOX family genes.


Comparative tolerability of paracetamol, aspirin and ibuprofen for short-term analgesia in patients with musculoskeletal conditions: results in 4291 patients.
The aim of this blinded, randomised, multicentre study was to compare the tolerability of aspirin, paracetamol and ibuprofen in common pain resulting from musculoskeletal conditions (MSC) in general practice with patients with other non-MSC pain conditions. Patients took aspirin, paracetamol (both up to 3g daily) or ibuprofen (up to 1.2g daily) for up to 7 days. The main outcome was the rate of significant adverse events (SGAE). Four thousand two hundred and ninety one patients with MSC were evaluable (1436 aspirin, 1423 paracetamol, 1432 ibuprofen) and 4101 (95.5%) were per-protocol. A group of 4342 patients included for other (non-MSC) mild to moderate pain conditions was used for comparison. In the MSC group, SGAE were reported by 20.5% of patients with aspirin, 17.0% with paracetamol and 15.0% with ibuprofen. Ibuprofen was statistically equivalent to paracetamol and better tolerated than aspirin (p <0.0001). Ibuprofen was associated with fewer digestive system AE (4.4%) than aspirin (8.6%, p<0.0001) and paracetamol (6.5%, p <0.02). The non-MSC group showed similar intertreatment differences, but experienced fewer SGAE. No serious digestive events were observed with any of the three treatments in either group. These results show that in patients with mild to moderate pain resulting from MSC, ibuprofen given in OTC doses for 6 days is as well tolerated as paracetamol and better tolerated than aspirin.


Role of VAMP-2, VAMP-7, and VAMP-8 in constitutive exocytosis from HSY cells.
We evaluated the role of VAMP-2/synaptobrevin, VAMP-7/TI-VAMP, and VAMP-8/endobrevin in exocytic pathways of HSY cells, a human parotid epithelial cell line, by coexpressing these VAMP proteins tagged with green fluorescent protein (GFP) and human growth hormone (hGH) as a secretory cargo. Exocytosis of hGH was constitutive and the fluorescent signal of hGH-GFP was observed in the Golgi area and small vesicles quickly moving throughout the cytoplasm. The cytoplasmic vesicles containing hGH overlapped well with VAMP-7-GFP, but did so scarcely with VAMP-2-GFP or VAMP-8-GFP. However, when the vesicle transport from the trans-Golgi network to the plasma membrane was arrested by incubation at 20 degrees C for 2 h and then released by warming up to 37 degrees C; VAMP-2-GFP and hGH were clearly colocalized together in small cytoplasmic vesicles. Neither VAMP-7-GFP nor hGH-GFP was colocalized with LAMP-1, a marker for lysosomes and late endosomes. These results suggest that (1) VAMP-2 can be one of the v-SNAREs for constitutive exocytosis; (2) VAMP-7 is involved in the constitutive exocytosis as a slow, minor v-SNARE, but not in the lysosomal transport; and (3) VAMP-8 is unlikely to be a v-SNARE for constitutive exocytosis in HSY cells.


Activation of a nonclassical NKT cell subset in a transgenic mouse model of hepatitis B virus infection.
NKT cells are specialized cells of the immune system that bear both T cell and NK cell markers. Classical NKT cells display TCRs of restricted heterogeneity (Valpha14-Jalpha281) and recognize lipid antigens (e.g., alpha-galactosyl ceramide) presented by nonpolymorphic CD1 molecules. Recently, other nonclassical NKT subsets have been recognized, including NKT cells not reactive with CD1d-alpha-galactosyl ceramide complexes. The biological functions of these cells are unknown. Here, we show that nonclassical NKT cells that are CD1d restricted but nonreactive to alpha-GalCer are activated in response to hepatocytes expressing hepatitis B viral antigens in a transgenic mouse model of acute hepatitis B virus infection. Our results document the first in vivo function for such nonclassical NKT cells and suggest a role for these cells in the host response to HBV infection.


Comparing bivariate reaction norms among species: time and size at metamorphosis in three species of Hyla (Anura: Hylidae).
Univariate and bivariate methods for comparing norms of reaction among species are discussed and illustrated with an example using North American hylid treefrogs. Norms of reaction for size at metamorphosis (SM) and length of larval period (LP) were compared among treefrog species raised at different food levels (Hyla cinerea vs H. gratiosa) and at different temperatures (H. cinerea vs H. gratiosa vs H. squirella). Hyla cinerea and Hyla gratiosa show parallel norms of reaction across food levels and temperatures. Across temperatures, H. squirella shows a much smaller change in SM relative to change in LP than do H. cinerea and H. gratiosa. This difference in shape of reaction norms may reflect different histories of selection resulting from these species' use of different larval habitats.


Seasonal variations in different forms of pokeweed antiviral protein, a potent inactivator of ribosomes.
Pokeweed antiviral proteins enzymatically inactivate the 60 S subunit of eucaryotic ribosomes in cell-free preparations. Three different species of the enzyme can be isolated from spring leaves, summer leaves, and seeds of pokeweed. Sequence analyses of the NH2-terminal residues show that pokeweed antiviral protein, isolated from spring leaves and seeds, are homologous and differ in 11 of the 28 residues compared. Ricin contains a polypeptide (ricin A chain) that has functional similarities to pokeweed antiviral protein, yet the sequences of the pokeweed proteins show little similarity with ricin A chain. Ricin B chain is responsible for helping ricin A chain across the plasma membrane; since pokeweed antiviral has no counterpart to ricin B chain, it is not nearly as cytotoxic as ricin. However, when pokeweed antiviral protein was covalently coupled to ricin B chain, a cytotoxic species was formed. Pokeweed antiviral protein fails to interact noncovalently with the ricin B chain to produce a cytotoxic species equivalent in function to ricin.


Changing scenario of malaria in central India, the replacement of Plasmodium vivax by Plasmodium falciparum (1986-2000).
Since 1986, we have been studying the changing epidemiology of malaria in a forest belt of Mandla, which has the highest number of malaria cases in central India (Madhya Pradesh) to define the epidemiological characteristics of the infection with each Plasmodium species in different seasons of the year. Our long-term objective was to determine the dynamics of Plasmodium vivax vs.P. falciparum infections. Five villages underwent fortnightly surveillance of fever cases. Drugs were distributed within 24 h after results of blood smears became available as per Indian National Anti-Malaria Programme. Indoor resting mosquitoes were also collected fortnightly. The only two Plasmodium species encountered were P. vivax and P. falciparum in both children and adults. Relatively more malaria infections were recorded in children (< or =14 years) than adults (>14 years) (chi2=89.94, P<0.00001). However, there were significant falling trends in P. vivax from 1986 to 2000 in both age groups (< or =14 years from 63 to 13, P<0.0001 and >14 years from 84 to 7, P<0.0001). The indoor resting density of Anopheles culicifacies, an efficient vector resistant to both dichlorodiphenyltrichloroethane (DDT) (4%) and hexachlorocyclohexane (HCH) (0.4%), was very high throughout this period in all villages (52.35 +/- 31.8, range 5-200 per man hour). Anopheles fluviatilis was present in small numbers 0.78 +/- 1.24 (range 0-7 per man hour). Major contributors of the changing epidemiology of malaria in this area are changing drug sensitivity along with insecticide sensitivity.


Possible disrupted biological movement processing in Developmental Coordination Disorder.
There is emerging evidence that the Mirror Neuron System (MNS) might contribute to the motor learning difficulties characteristic of Developmental Coordination Disorder (DCD). This study aimed to identify whether MNS activity differed between children with and without DCD during action observation, action execution and during a non-action baseline. Electroencephalography (EEG) was used to measure mu rhythm (a proxy for MNS activation) in 8-12-year-old children either with (n = 20) or without (n = 19) a diagnosis of DCD. The mu rhythm was recorded at rest and during five experimental conditions: (1) observation of gross motor and (2) fine motor actions; (3) execution of gross motor and (4) fine motor actions; and (5) non-biological movement. To address whether potential co-occurring traits of other neurodevelopmental conditions were associated with differences in mu rhythm, parents reported their child's attention and social communication skills. Mixed and repeated measure ANOVAs were conducted to examine differences in mu desynchronization and mu power respectively. The non-DCD group showed greater mu rhythm desynchronization than children with DCD (i.e., more MNS activity), with both groups demonstrating increasing desynchronization from observation of fine actions to execution of gross actions. However, we also found that the children with DCD had less mu power during the non-biological movement condition than the non-DCD children, although mu power did not differ between groups during the resting condition. Correlations between mu desynchronization and children's attention and motor skills showed that poorer attention and motor abilities were associated with reduced MNS activity. Compared to children without DCD, the MNS in children with DCD did not distinguish between biological and non-biological movement. It is possible that the reduced specificity of the MNS in children with DCD is an underlying factor in the motor impairments observed in the disorder. The differential MNS activity could reflect broader atypical activity in perceptual networks that feed into the MNS in DCD.


Biomechanical comparison of lateral collateral ligament reconstruction with and without additional internal bracing using a three-dimensional elbow simulator.
Although an additional internal bracing significantly increases stability in a repair of the lateral ulnar collateral ligament, it remains unclear whether it also does in reconstruction. Aim of this study was to implement a three-dimensional elbow simulator for testing posterolateral rotatory instability. We hypothesized that (1) reconstruction with and without internal bracing is comparable in biomechanical properties, and (2) there would be higher load-to-failure with internal bracing. Posterolateral rotatory instability was tested by imitating the lateral pivot shift test in 16 elbows. Valgus and supination torques were simultaneously increased stepwise up to 1.2 Nm. Specimens were tested at 30°, 60°, 90°, and 120° elbow flexion with an intact lateral collateral ligament complex, dissected complex, and after reconstruction with or without internal bracing. Outcome measures included joint gapping, laxity, and load to failure. With the implemented elbow simulator no significant difference was observed for gapping or laxity between both treatment groups. Comparing treatment and native ligament, gapping was reduced, especially with increased elbow flexion. Laxity was also reduced at some flexion angles. The mean load-to-failure was 8.1 ± 2.7 Nm without and 9.6 ± 3.6 Nm with internal bracing (P = 0.645). Both treatments were comparable in biomechanical properties but did not fully restore the native state. Although the additional augmentation of the LUCL reconstruction tends to increase the maximum load to failure, this difference was not statistically significant. Still, reconstruction with internal bracing seems to be a reasonable option in selected primary reconstructions. It could also be useful in revision reconstruction.


Na+,K+-ATPase interaction with a brain endogenous inhibitor (endobain E).
Na+,K+-ATPase activity of rat brain synaptosomal membranes was evaluated in the presence of an inhibitory fraction II-E (termed endobain E), isolated by gel filtration and anionic exchange HPLC of a rat brain soluble fraction. We studied endobain E aging, analyzed its inhibitory potency in the absence or presence of ouabain as well as its ability to block high affinity [3H]ouabain binding to cerebral cortex membranes. Similar loss of endobain E activity was observed when samples were stored either dried or in solution. Endobain E fraction inhibited synaptosomal membrane Na+,K+-ATPase activity in a concentration-dependent manner and the slope of the corresponding curve strongly resembled that of ouabain. Assays performed in the presence of endobain E and ouabain indicated that the inhibitory effect was additive or less than additive, depending on their respective concentrations during preincubation and/or incubation. High affinity [3H]ouabain binding to cerebral cortex membranes proved concentration-dependent from 0.10 to 0.50 mg protein per ml; binding inhibition by endobain E was independent of protein concentration within the above range. [3H]ouabain binding inhibition by endobain E was concentration-dependent over a 10-fold range, an effect similar to that found for Na+,K+-ATPase inhibition. The extent of endobain E effect on Na+,K+-ATPase inhibition was much higher (90-100%) than that on [3H]ouabain binding blockade (50%). Findings suggest some type of interaction between endobain E and ouabain inhibitory mechanisms and favour the view that the former behaves as an endogenous ouabain.


Translation in vitro of RNA from the human malarial parasite Plasmodium falciparum.
RNA has been isolated from the human malarial parasite, Plasmodium falciparum, and translated in vitro using two systems: rabbit-reticulocyte lysate and Xenopus laevis oocytes. Polypeptides are synthesized that are characteristic of the parasite isolate and the developmental stage from which the RNA was purified. Many of these polypeptides are precipitated by antibody from an immune monkey. Analysis of the primary structure of these polypeptides should give the information necessary to produce a vaccine against this parasite.


Postnatal changes in Bax-immunoreactivity and apoptosis of the rat trigeminal primary neurons.
The developmental changes of Bax protein-like immunoreactivity (Bax-ir) and naturally occurring cell death were investigated in the trigeminal primary neurons of rats, in the first 2 postnatal weeks. At 1 day postpartum, the trigeminal primary neurons exhibited intense cytoplasmic Bax-ir. A densitometric analysis indicated >95% of cells exhibited an ir-density (the staining intensity as compared to the background level) higher than 1.5. By 2 weeks the ir-density significantly decreased with >95% lower than 1.5, that was similar to the adult level. A nick-end labeling method revealed DNA fragmentation in apoptotic trigeminal primary neurons in newborn rats. During the first 11 days the percentage of apoptotic cells was highly correlated to the postnatal days with a regression line y = -0.083x + 0.877 (r = 0.976). A possible role of Bax in neonatal rat primary neurons is discussed.


The pharmacology of prostaglandin-like substances released from guinea-pig lungs during anaphylaxis.
The pharmacology of the prostaglandins (PGs) and thromboxanes (Txs) released from immunologically challenged guinea-pig lungs is related to their roles in the anaphylactic response. 6-oxo-PGF1alpha probably contibutes substantially to bronchoconstriction during anaphylaxis. TxB2 may contribute to the anaphylactic response by increasing SRA-A release and by stimulating leucotaxis. The 15-oxo metabolites of PGE2 and PGF2alpha are rather weak spasmogens, but might modify respiratory muscle contractions and pulmonary vascular resistance. The 15-oxo 13,14 dihydro metabolites of PGE2 PGF2alpha and TxB2 were inactive in the systems studied, suggesting an important inactivating role for the 13:14 reductase enzyme.


New Daily Persistent Headache (NDPH): Unraveling the Complexities of Diagnosis, Pathophysiology, and Treatment.
The current article aims to provide an overview of new daily persistent headache (NDPH), with a particular emphasis on its pathophysiology, evaluation, and current treatment options. NDPH is an uncommon and heterogeneous condition associated with various comorbidities and is of great significance due to its prolonged duration and high severity. Variable causes and clinical aspects of NDPH may reflect differences in its underlying pathophysiological mechanisms, including genetics, environmental triggers, neuroinflammation, and brain changes. When assessing a patient with NDPH, potential triggers, past medical history, and differential diagnosis should be carefully considered. Non-pharmacological interventions aimed to improve diet, sleep patterns, and reduce consumption of caffeine and alcohol are recommended for all patients. Nerve blockade and nerve stimulation seem to be more efficacious in children than adults. Antiviral medications and neuroinflammation-targeting treatments may be helpful, particularly, when an infectious disease or severe inflammation is suspected. NDPH patients with concurrent affective disorders may benefit from treatment with serotonin reuptake inhibitors, serotonin/norepinephrine reuptake inhibitors, or benzodiazepines. Cerebrospinal-fluid-lowering medications may be useful for headaches started with a thunderclap or a Valsalva maneuver. Possible treatments for refractory NDPH include intravenous ketamine or lidocaine, onabotulinumtoxinA, and calcitonin gene-related peptide antibodies. Considering the variety of NDPH, it is critical to properly screen patients for correct diagnosis. Proper identification of potential mimics may enable precise therapy opportunities, yet there is no gold standard treatment for NDPH. Further well-designed studies are needed to elucidate the underlying mechanisms and develop effective treatment strategies for NDPH.


The deubiquitylating enzyme USP44 counteracts the DNA double-strand break response mediated by the RNF8 and RNF168 ubiquitin ligases.
Protein recruitment to DNA double-strand breaks (DSBs) relies on ubiquitylation of the surrounding chromatin by the RING finger ubiquitin ligases RNF8 and RNF168. Flux through this pathway is opposed by several deubiquitylating enzymes (DUBs), including OTUB1 and USP3. By analyzing the effect of individually overexpressing the majority of human DUBs on RNF8/RNF168-mediated 53BP1 retention at DSB sites, we found that USP44 and USP29 powerfully inhibited this response at the level of RNF168 accrual. Both USP44 and USP29 promoted efficient deubiquitylation of histone H2A, but unlike USP44, USP29 displayed nonspecific reactivity toward ubiquitylated substrates. Moreover, USP44 but not other H2A DUBs was recruited to RNF168-generated ubiquitylation products at DSB sites. Individual depletion of these DUBs only mildly enhanced accumulation of ubiquitin conjugates and 53BP1 at DSBs, suggesting considerable functional redundancy among cellular DUBs that restrict ubiquitin-dependent protein assembly at DSBs. Our findings implicate USP44 in negative regulation of the RNF8/RNF168 pathway and illustrate the usefulness of DUB overexpression screens for identification of antagonizers of ubiquitin-dependent cellular responses.


The effect of the oxic cell sensitizer, beta-ara A on human fibroblasts in plateau and in exponential growth phases.
Before an oxic cell sensitizer such as beta-ara A (a DNA-dependent DNA polymerase inhibitor) can be used in cancer treatment, it is essential to know both the influence of this type of drug on certain critical normal tissues and the role of proliferation kinetics in the radiosensitizing capacity. The biological system chosen for this in vitro study was the human fibroblast cell line HF19. Cells were studied in plateau phase and in the exponential growth phase. Cells were incubated with beta-ara A for 7 hr (1 hr before and 6 hr after irradiation). beta-ara A was extremely toxic to growing cells (concentrations ranging from 200 to 1000 microM), but no detectable effect was found on plateau-phase cells (up to 4000 microM). However, for a given drug concentration, the radiosensitizing effect (Sensitizing Enhancement Ratio SER) was very similar for growing and plateau phase cells (SER measured with Ds ratio was about 1.7 for a concentration of 500 microM). The enhancement ratio depended on the radiation dose; it was relatively higher for low doses. This can be explained by a differential effect of the drug on the alpha and beta components of the survival curve. Only the alpha component was increased.


Systems for producing precise movements of a joint over a wide range of speeds and displacements for tests of a static-position sense.
This report describes 3 types of apparatus that were used to produce precise movements of a joint over a wide range of speeds and angles. The designs feature an ability for ultra slow rotation of the joint (fractions of a degree per min) with a minimum of extraneous cues. Two designs use servo-controlled DC motors configured as velocity servos and a third design uses a galvanometer motor configured as a position servo. Originally designed for use with humans in studies of proprioception with the ankle and two joints of the index finger (the metacarpophalangeal joint and proximal interphalangeal joint), the apparatuses should be useful in a variety of applications where precise control of velocity and position is needed.


Impacts of exogenous mineral silicon on cadmium migration and transformation in the soil-rice system and on soil health.
Exogenous silicon has been shown to enhance plant growth and alleviate heavy metals toxicity, but the regulation mechanism of silicon on cadmium migration and transformation in the soil-rice system is still unclear, which is worth further study. In this study, a pot experiment was carried out to explore the influence of different doses (0, 1 and 5%) of mineral silicon on soil properties, nutrient availability, rice growth, soil enzyme activities, Cd bioavailability, and uptake and accumulation of Cd in high-accumulating (H) and low-accumulating (L) rice cultivars grown in contaminated soils. Results showed that mineral-Si treatment could increase the total biomass and grain yield, with an increased rate of 17.7-27.3% and 14.7-19.1% for H; while 26.2-33.4% and 21.3-30.3% for L. Compared with non-mineral-Si treatment, the soil EX-Cd decreased by 3.9-13.3% (H) and 2.3-10.7% (L). Additionally, the Cd content in rice grain was significantly declined by 29.5-31.3% (H) and 34.9-35.2% (L). Mineral-Si enhanced urease, sucrase, and neutral phosphatase activities in both cultivars, but suppressed catalase activity in H. A selective change in bacterial community structure was observed under mineral-Si treatment, however, the bacterial community remained stable, suggesting that the mineral-Si had no adverse effect on the microbial community. The positive response of soil enzymes activities, rice growth and the overall stabilization of microbial environment for mineral-Si addition to the Cd contaminated soils indicated that mineral-Si could mitigate the risk of Cd and well maintain the soil health, proving it to be eco-friendly and low-cost amendment for soils remediation.


Usefulness of high-dose intravenous human immunoglobulins treatment for refractory recurrent pericarditis.
The management of refractory recurrent pericarditis is challenging. Previous clinical reports have noted a beneficial effect of high-dose intravenous human immunoglobulins (IvIgs) in isolated and systemic inflammatory disease-related forms. In this article, we analyzed retrospectively our clinical experience with IvIg therapy in a series of clinical cases of pericarditis refractory to conventional treatment. We retrospectively analyzed 9 patients (1994 to 2010) with refractory recurrent pericarditis, who received high-dose IvIg as a part of their medical treatment. Nonsteroidal anti-inflammatory drugs (NSAIDs), steroids, or colchicine treatment was not discontinued during IvIg treatment. No patients had a history of autoimmune or connective tissue diseases. During an average period of 11 months from the first recurrence, patients had experienced a mean of 5 relapses before the first IvIg treatment. In 4 cases, patients showed complete clinical remission with no further relapse after the first IvIg cycle. Two patients experienced a single minor relapse, responsive to short-term nonsteroidal anti-inflammatory drugs. In 2 patients, we performed a second cycle of IvIg after a recurrence of pericarditis, with subsequent complete remission. One patient did not respond to 3 cycles of IvIg and subsequently underwent pericardial window and long-term immunosuppressive treatment. No major adverse effect was observed in consequence of IvIg administration in all the cases. In conclusion, although IvIg mode of action is still poorly understood in this setting, this treatment can be considered as an option in patients with recurrent pericarditis refractory to conventional medical treatment and, in our small series, has proved to be effective in 8 of 9 cases.


Speciation analysis of arsenic in terrestrial plants from arsenic contaminated area.
Arsenic speciation analysis was carried out in plants collected from arsenic contaminated area. Two plant species were chosen for the investigation: Reed Grass (Calamagrostis arundinacea) and Lady Fern (Athyrium filix-femina). To characterize arsenic species several different extraction procedures were applied including enzymatic extraction and extraction using surfactant solution (SDS). Two-step sequential extraction (water+SDS) that assures the highest extraction efficiency was applied to extract arsenic species from plant material. HPLC with anion-exchange column was used to separate extracted arsenic compounds and ICP-MS was applied for quantitative arsenic determination after species separation.


Match running performance profiles of amputee football players at the national level.
Even though running performance and positional profiles in football are well described, amputee football (AF) has different characteristics of the movement, pitch dimensions, and time played. There is a gap in the scientific literature about positional profiles based on running performance in AF. This study aimed to investigate the differences between positions, the influence of the amputation level or defect of the lower limb (LD), the differences in running performance between halves, and the relationship with the final match result. Thirteen AF National Team players were monitored by Global Navigation Satellite System (GNSS), tracking 24 official international matches for 17 months. Values of top speed, peak acceleration, peak deceleration, average distance per minute, sprint mean speed, GPS load per minute, inertial load per minute (Gs load/min), number of sprints per minute, and a number of impacts per minute were analyzed for defenders (DEF), midfielders (MID) and forwards (FOR). Additional factors analyzed were amputation level (below the knee, low amputation-LA or over the knee, high amputation-HA or defect of the lower limb-LD) and the match's final result. Midfielders had significantly higher running performance parameters compared to other positions (greater top speed than DEF (+ 0.3 m/s; p < 0.001) and FOR (+ 0.2 m/s; p = 0.045), greater peak acceleration and deceleration than DEF (+ 0.5 m/s2 for both measures; p < 0.001) and FOR (+ 0.4 and + 0.3 m/s2; p < 0.001 and p = 0.036, respectively), greater GPS load/minute than DEF (+ 0.2 load/min; p = 0.001) and FOR (+ 0.3 load/min; p < 0.001), greater Gs load per minute than DEF (+ 2.7 load/min; p < 0.001) and FOR (+ 1.8 load/min; p < 0.001), greater number of impacts per minute than DEF (+ 0.2 n/min; p < 0.001) and FOR (+ 0.2 n/min; p < 0.001). Players with LD had significantly higher running performance than those with LA or HA. In the match's second half, a decrease in running performance was registered. The trend of running more when losing could be observed-AF players had higher running parameters in lost matches, but the differences were not statistically significant. Further research complied with contextual game analysis is needed to assess the running performance of AF players deeply.


High-performance liquid chromatography-tandem mass spectrometry in the identification and determination of phase I and phase II drug metabolites.
Applications of tandem mass spectrometry (MS/MS) techniques coupled with high-performance liquid chromatography (HPLC) in the identification and determination of phase I and phase II drug metabolites are reviewed with an emphasis on recent papers published predominantly within the last 6 years (2002-2007) reporting the employment of atmospheric pressure ionization techniques as the most promising approach for a sensitive detection, positive identification and quantitation of metabolites in complex biological matrices. This review is devoted to in vitro and in vivo drug biotransformation in humans and animals. The first step preceding an HPLC-MS bioanalysis consists in the choice of suitable sample preparation procedures (biomatrix sampling, homogenization, internal standard addition, deproteination, centrifugation, extraction). The subsequent step is the right optimization of chromatographic conditions providing the required separation selectivity, analysis time and also good compatibility with the MS detection. This is usually not accessible without the employment of the parent drug and synthesized or isolated chemical standards of expected phase I and sometimes also phase II metabolites. The incorporation of additional detectors (photodiode-array UV, fluorescence, polarimetric and others) between the HPLC and MS instruments can result in valuable analytical information supplementing MS results. The relation among the structural changes caused by metabolic reactions and corresponding shifts in the retention behavior in reversed-phase systems is discussed as supporting information for identification of the metabolite. The first and basic step in the interpretation of mass spectra is always the molecular weight (MW) determination based on the presence of protonated molecules [M+H](+) and sometimes adducts with ammonium or alkali-metal ions, observed in the positive-ion full-scan mass spectra. The MW determination can be confirmed by the [M-H](-) ion for metabolites providing a signal in negative-ion mass spectra. MS/MS is a worthy tool for further structural characterization because of the occurrence of characteristic fragment ions, either MS( n ) analysis for studying the fragmentation patterns using trap-based analyzers or high mass accuracy measurements for elemental composition determination using time of flight based or Fourier transform mass analyzers. The correlation between typical functional groups found in phase I and phase II drug metabolites and corresponding neutral losses is generalized and illustrated for selected examples. The choice of a suitable ionization technique and polarity mode in relation to the metabolite structure is discussed as well.


Natural History of Perianal Crohn's Disease: Long-term Follow-up of a Population-Based Cohort.
The natural history of perianal Crohn's disease (PCD) remains poorly described and is mainly based on retrospective studies from referral centers. The aim of this study was to assess the incidence, outcomes and predictors of the onset of PCD. All incident cases of patients diagnosed with possible CD were prospectively registered from 1994 to 1997 in Brittany, a limited area in France. At diagnosis, the clinical features of perianal disease were recorded. All patient charts were reviewed from the diagnosis to the last clinic visit in 2015. Among the 272 out of 331 incident CD patients followed up, 51 (18.7%) patients had PCD at diagnosis. After a mean follow-up of 12.8 years, 93 (34%) patients developed PCD. The cumulative probabilities of perianal CD occurrence were 22%, 29%, and 32% after 1 year, 5 years, and 10 years, respectively. The cumulative probabilities of anal ulceration were 14%, and 19% after 1 year and 10 years, respectively. Extraintestinal manifestations were associated with the occurrence of anal ulceration. The cumulative probabilities of fistulizing PCD were 11%, 16%, and 19% after 1 year, 5 years, and 10 years, respectively. Extraintestinal manifestations, rectal involvement and anal ulceration were predictors of fistulizing PCD. The cumulative probability of developing anal stricture was 4% after 10 years. PCD is frequently observed during CD, in approximately one-third of patients. These data underline the need for targeted therapeutic research on primary perianal lesions (proctitis, anal ulceration) to avoid the onset of fistulizing perianal disease.


Seed priming with proline improved photosystem II efficiency and growth of wheat (Triticum aestivum L.).
Proline can promote growth of plants by increasing photosynthetic activity under both non-stress and abiotic stress conditions. However, its role in non-stressed conditions is least studied. An experiment was conducted to assess as to whether increase in growth of wheat due to seed priming with proline under non-stress condition was associated with proline-induced changes in photosystem II (PSII) activity. Seeds of four wheat varieties (S-24, Sehar-06, Galaxy-13, and Pasban-90) were primed with different concentrations of proline (0, 5, 15 and 25 mM) for 12 h and allowed to grow under normal conditions. Biomass accumulation and photosynthetic performance, being two most sensitive features/indicators of plant growth, were selected to monitor proline modulated changes. Seed priming with proline increased the fresh and dry weights of shoots and roots, and plant height of all four wheat varieties. Maximum increase in growth attributes was observed in all four wheat varieties at 15 mM proline. Maximum growth improvement due to proline was found in var. Galaxy-13, whereas the reverse was true for S-24. Moreover, proline treatment changed the Fo, Fm, Fv/Fo, PIABS, PITot in wheat varieties indicating changes in PSII activity. Proline induced changes in energy fluxes for absorption, trapping, electron transport and heat dissipation per reaction center indicated that var. Galaxy-13 had better ability to process absorbed light energy through photosynthetic machinery. Moreover, lesser PSII efficiency in var. S-24 was due to lower energy flux for electron transport and greater energy flux for heat dissipation. This was further supported by the fact that var. S-24 had disturbance at acceptor side of PSI as reflected by reduction in ΔVIP, probability and energy flux for electron transport at the PSI end electron acceptors. Seed priming with proline improved the growth of wheat varieties, which depends on type of variety and concentration of proline applied. Seed priming with proline significantly changed the PSII activity in wheat varieties, however, its translation in growth improvement depends on potential of processing of absorbed light energy by electron acceptors of electron transport chain, particularly those present at PSI end.


Dried Blood Spots in Neonatal Studies: A Computational Analysis for the Role of the Hematocrit Effect.
Dried blood spot (DBS) microsampling is extensively employed in newborn screening (NBS) and neonatal studies. However, the impact of variable neonatal hematocrit (Ht) values on the results can be a source of analytical error, and the use of fixed Ht for calibration (Htcal) is not representative of all neonatal subpopulations. A computational approach based on neonatal demographics was developed and implemented in R® language to propose a strategy using correction factors to address the Ht effect in neonatal DBS partial-spot assays. A rational "tolerance level" was proposed for the Ht effect contribution to the total analytical error and a safe Ht range for neonatal samples, where the correction of concentrations can be omitted. Furthermore, an "alert zone" for a false positive or negative result in NBS was proposed, where the Ht effect has to be considered. Results point toward the use of Htcal values closely representative of populations under analysis and an acceptable level of percentage relative error can be attributed to the Ht effect, diminishing the probability of correction. Overall, the impact of the Ht effect on neonatal studies is important and future work may further investigate this parameter, correlated to other clinical variables potentially affecting results.


Update: Salmonella enteritidis infections and shell eggs--United States, 1990.
From January through October 1990, state health departments reported 49 outbreaks of Salmonella enteritidis (SE) in the United States to CDC. This report summarizes three SE outbreaks in 1990 that were associated with shell eggs.


Oxidation of cholesteryl linoleate by human monocyte-macrophages in vitro.
The amount of cholest-5-en-3 beta,7 beta-diol (CD) was significantly higher in cultures of human monocytes incubated with cholesteryl linoleate-bovine serum albumin (CL/BSA) artificial lipoproteins than in no-cell control incubations of CL/BSA. CD production by monocytes was almost completely inhibited by the radical scavengers butylated hydroxytoluene (BHT), probucol, and alpha-tocopherol, and was partially inhibited by the metal chelator EDTA. The production of CD was accompanied by decrease in linoleic acid. CD amounts were negligible in incubations of monocytes with cholesteryl oleate/BSA (CO/BSA) or cholesterol/BSA (C/BSA). Ability to produce CD from CL/BSA appeared to increase with age in culture of human monocyte-macrophages. Considerable variations were observed in the CD production from CL/BSA by monocytes from different blood donations. Higher levels of CD production appeared more common with monocytes from men than from women. The significance of these results in the context of human atherosclerosis is discussed.


Selective facilitatory effect of vasoactive intestinal polypeptide (VIP) on muscarinic firing in vesical ganglia of the cat.
VIP immunoreactivity was identified in nerve fibers and in 10-13% of the neurons in pelvic and bladder ganglia of the cat. Ninety percent of the VIP positive neurons contained acetylcholinesterase. VIP immunoreactivity was not altered in decentralized ganglia 1 week to 8 months after transection of the pelvic and hypogastric nerves indicating that VIP fibers arose from neurons within the peripheral nervous system. The intra-arterial administration of VIP (1-50 micrograms/kg) enhanced the postganglionic discharge elicited by the muscarinic agonist, acetyl-beta-methylcholine, but did not alter the postganglionic firing elicited by the nicotinic agonist, tetramethylammonium or by electrical stimulation of preganglionic axons in the pelvic nerve. VIP did not elicit a postganglionic discharge in untreated ganglia, but did evoke a prolonged discharge in ganglia treated with an irreversible anticholinesterase agent, 217AO. This discharge was not affected by hexamethonium but was blocked by atropine. VIP suppressed the muscarinic inhibition of ganglionic transmission produced by acetyl-beta-methylcholine without altering the response to other inhibitory agents (norepinephrine, leucine-enkephalin and gamma-aminobutyric acid (GABA). VIP (0.1-0.3 micrograms/kg) also had a direct inhibitory effect on bladder smooth muscle. These findings raise the possibility that intraganglionic pathways containing VIP may exert a selective modulatory influence on muscarinic transmission in vesical parasympathetic ganglia.


Toxicogenomic study in rat thymus of F1 generation offspring following maternal exposure to silver ion.
Male and female rats (26-day-old) were exposed to 0.0, 0.4, 4 or 40 mg/kg body weight silver acetate (AgAc) in drinking water for 10 weeks prior to and during mating. Sperm-positive females remained within their dose groups and were exposed to silver acetate during gestation and lactation. At postnatal day 26, the effect of silver ions on the developing F1 generation rat thymus was evaluated at the transcriptional level using whole-genome microarrays. Gene expression profiling analyses identified a dozen differentially expressed genes (DEGs) in each dose group using a loose criterion of fold change (FC) >1.5 and unadjusted p < 0.05, regardless of whether the analysis was conducted within each gender group or with both gender groups combined. No dose-dependent effect was observed on the number of DEGs. In addition, none of these genes had a false discovery rate (FDR) <0.05 after correction for multiple testing. These results in combination with the observation that thymus-to-body-weight ratios were not affected and no histopathological abnormalities were identified indicate that in utero exposure to silver ions up to 26.0 mg/kg (equivalent to 40.0 mg/kg silver acetate) did not have an adverse effect on the developing thymus.


Clinical and behavioural features of SYNGAP1-related intellectual disability: a parent and caregiver description.
SYNGAP1-related intellectual disability (ID) is a recently described neurodevelopmental disorder that is caused by pathogenic variation in the SYNGAP1 gene. To date, the behavioural characteristics of this disorder have mainly been highlighted via the prevalence of existing diagnoses in case series. We set out to detail the behavioural features of this disorder by undertaking interviews with those who have a child with SYNGAP1-related ID to allow them to describe their child's behaviour. We conducted 27 semi-structured interviews with parents and caregivers which covered basic information (e.g., age, gender), family history, perinatal history, past medical history, developmental history, epilepsy, behavioural history, and a general description of their child's behaviour. Using a mixed quantitative and qualitative approach, the responses from the parents indicated that those with SYNGAP1-related ID showed high rates of autism spectrum disorder (52%), difficulties with fine and gross motor skills, delays in language development, and a high prevalence of epilepsy (70%). A qualitative analysis highlighted their general behaviour affected the themes of daily living skills, distress-related behaviours, emotional regulation, difficulties with change, a lack of danger awareness, and sensory differences. Sensory features described involved auditory, visual, tactile, gustatory, and proprioceptive themes. Our findings and behavioural descriptions provide important insights as well as implications for the diagnosis and care of those with SYNGAP1-related ID.


Realizing Efficient EMG-Based Prosthetic Control Strategy.
As an integral part of the body, the limb poses dexterous and fine motor grasping and sensing capabilities that enable humans to effectively communicate with their environment during activities of daily living (ADL). Hence, limb loss severely limits individuals' ability especially when they need to perform tasks requiring their limb functions during ADL, thus leading to decreased quality of life. To effectively restore limb functions in amputees, the advanced prostheses that are controlled by electromyography (EMG) signal have been widely investigated and used. Since EMG signals reflect neural activity, they would contain information on the muscle activation related to limb motions. Pattern recognition-based myoelectric control is an important branch of the EMG-based prosthetic control. And the EMG-based prosthetic control theoretically supports multiple degrees of freedom movements that allows amputees to intuitively manipulate the device. This chapter focuses on EMG-based prosthetic control strategy that involves utilizing intelligent computational technique to decode upper limb movement intentions from which control commands are derived. Additionally, different techniques/methods for improving the overall performance of EMG-based prostheses control strategy were introduced and discussed in this chapter.


DNA hypomethylation in Morris hepatomas.
The 5-methylcytosine (m5C) content of DNAs from Morris hepatomas of varying growth rates and from normal liver was analyzed. DNA methylation in all hepatomas studied was found to be 20-45% less than in normal liver. This result was confirmed independently by restriction endonuclease (Hpa II and Msp I) analysis. While these results agreed with recent literature data suggesting hypomethylation of DNA from some neoplastic sources, no correlation was observed between the extent of DNA hypomethylation and the growth rates of the tumors.


Will nuclear polluted seafood stop selling in the blockchain-enabled market? Lessons from government punishment and social cognition for retailer's selling.
In April 2021, Japan's decision to dump nuclear wastewater into the ocean has raised worldwide attention. Therefore, to focus on seafood safety from firm and government in this event, we construct a game model to explore the technology-enabled ways to resolve conflict from domestic product and polluted product. Our analysis reveals the potential equilibrium strategy for the domestic product only and two types of products, respectively. Moreover, from the perspective of government punishment, the result shows the existing motivation of polluted product in the market. Finally, we also investigate the five aspects to find the changes of market share for domestic product, including customer choice, product freshness, market uncertainty, geographical distance, and shoddy product, respectively. In summary, this research provides management implications to resolve conflict between two types of products and to realize the multi-party balance of interests and technology-enabled value.


Mediterranean fruit fly, Ceratitis capitata (Wiedemann), mitochondrial DNA: genes and secondary structures for six t-RNAs.
The polymerase chain reaction was used to amplify six mitochondrial t-RNAs for Ala, Arg, Asn, Ser, Glu and Phe between genes for mitochondrial NADH dehydrogenases 3 and 5. With respect to Drosophila yakuba the gene order and direction of transcription is completely conserved. Analysis of secondary structure shows complete conservation of the anticodon loops but a number of differences in the dihydrouridine and T psi C loops with respect to Drosophila. However, differences are such that tertiary interactions that stabilize stacking are preserved. The use of the reported sequence in combination with PCR to explore population variability is discussed.


Feasibility of a Habit Formation Intervention to Delay Frailty Progression Among Older African Americans: A Pilot Study.
Older African Americans are at high risk for becoming frail in later life. Interventions can reverse or delay frailty, yet African Americans have largely been excluded from such research. Many interventions are also time- and resource-intensive, and thus inaccessible to socially disadvantaged older African Americans. We evaluated the feasibility of a low-dose frailty prevention intervention integrated with primary care among 60 community-dwelling, prefrail older African Americans aged 55+ recruited from a primary care clinic. We conducted a 2-arm randomized control trial. Participants were assigned to a 4-session intervention (1 session per month), delivered by an occupational therapist, or enhanced usual care. Feasibility criteria were set a priori at 75% for participant retention (including attrition due to death/hospitalization), 80% for session engagement, 2 participants/week for mean participant accrual, and 90% for program satisfaction. Participants were 65% female with an average age of 76.6 years, 51.7% lived alone and 39.1% reported <$10,000 in yearly income. Feasibility metrics were met. The study recruited, a mean of, 2.5 participants per week and retained 75% of participants who attended 95% of scheduled sessions. The mean satisfaction score was 29.75 (range = 0-32; SD .25). Changes in physical activity and dietary habit formation as well as changes in secondary outcomes were largely in the expected direction. The intervention was feasible to deliver. Qualitative findings from exit interviews suggested changes to the program dose, structure, and content that could improve it for future use.


Applying hierarchical task analysis to medication administration errors.
Medication use in hospitals is a complex process and is dependent on the successful interaction of health professionals functioning within different disciplines. Errors can occur at any one of the five main stages of prescribing, documenting, dispensing or preparation, administering and monitoring. The responsibility for the error is often placed on the nurse, as she or he is the last person in the drug administration chain whilst more pressing underlying causal factors remain unresolved. This paper demonstrates how hierarchical task analysis can be used to model drug administration and then uses the systematic human error reduction and prediction approach to predict which errors are likely to occur. The paper also puts forward design solutions to mitigate these errors.


Characterizing the impact of minor cannula design modification.
Bench evaluation of the hydrodynamic behavior of venous cannulas is a valuable technique for the analysis of their performance during cardiopulmonary bypass (CPB). The aim of this study was to investigate the effect of the internal diameter of the extracorporeal connecting tube of venous cannulas on flow rate (Q), pressure drop (delta P), and cannula resistance (delta P/Q²) values, using a computer assisted test bench. An in vitro circuit was set up with silicone tubing between the test cannula encased in a movable reservoir, and a static reservoir. The delta P, defined as the difference between the drainage pressure and the preload pressure, was measured using high-fidelity Millar pressure transducers. Q was measured using an ultrasonic flowmeter. Data display and data recording were controlled using virtual instruments in a stepwise fashion. The 27 F smartcanula® with a 9 mm connecting tube diameter showed 17% less resistance compared to that with an 8 mm connecting tube diameter. Q values were 7.22±0.1 and 7.81±0.04 L/min for cannulas with 8 mm and 9 mm connecting tube diameters, respectively. The delta P/Q² ratio values were 72% lower for the Medtronic cannula with a 9 mm connecting tube diameter compared to that with an 8 mm connecting tube diameter. Q values for the Medtronic cannula were 3.94±0.23 and 6.58±0.04 L/min with 8 mm and 9 mm connecting tube diameters, respectively. The 27 F smartcanula® showed 13% more flow rate compared to the 28 F Medtronic cannula using the unpaired Student t-test (p<0.0001). Our results demonstrated that Q was increased but delta P and delta P/Q² values were significantly decreased when the connecting tube diameter was increased for venous cannulas. The connecting tube diameter significantly affected the resistance to liquid flow through the cannula. Smartcanulas® outperform Medtronic cannulas.


Molecular cloning and expression of TLR in the Eisenia andrei earthworm.
Toll-like receptors (TLRs) play an important role in defense responses to pathogens in invertebrates. Here we characterize the first TLR isolated from an oligochaete annelid, namely, Eisenia andrei (EaTLR) and show its expression pattern. The full-length EaTLR cDNA consists of 2615 bp encoding a putative protein of 675 amino acids. The predicted amino acid sequence comprises of an extracellular domain containing 31 amino acid signal peptide and seven leucine-rich repeats (LRR), capped with cysteine-rich N- and C-terminal LRRs followed by a transmembrane domain and cytoplasmic Toll/IL-1R domain (TIR). TIR domains of twenty individual earthworms were sequenced and the variability suggesting the presence of a high number of TLR genes in the genome of E. andrei was observed. Phylogenetic analysis revealed the highest similarity of EaTLR with polychaete annelid, Capitella teleta and TLRs of mollusks and echinoderms. Finally, the highest constitutive expression of EaTLR was observed in the digestive tract. Gene expression was significantly increased in coelomocytes of E. andrei after the challenge with Gram-positive bacteria.


The Trend of Age-Group Effect on Prognosis in Differentiated Thyroid Cancer.
Age has been included in various prognostic scoring systems for differentiated thyroid cancer (DTC). The aim of this study is to re-examine the relationship between age and prognosis by using Surveillance, Epidemiology, and End Results (SEER) population-based database. We identified 51,061 DTC patients between 2004 and 2012. Patients were separated into 10-year age groups. Cancer cause-specific survival (CSS) and overall survival (OS) data were obtained. Kaplan-Meier and multivariable Cox models were built to analyze the outcomes and risk factors. Increasing age gradient with a 10-year interval was associated with the trend of higher proportions for male gender, grade III/IV and summary stage of distant metastases. Both CSS and OS continued to worsen with increasing age, being poorest in in the oldest age group (≥71); multivariate analysis confirmed that CSS continued to fall with each age decade, significantly starting at 60 years (HR = 7.5, 95% 1.0-54.1, p = 0.047) compared to the young group (≤20). Similarly, multivariate analysis suggested that OS continued worsening with increasing age, but starting at 40 years (HR = 3.7, 95% 1.4-10.1, p = 0.009) compared to the young group. The current study suggests that an age exceeding 60 years itself represents an unfavorable prognostic factor and high risk for cancer-specific death in DTC.


Microbial profile of burn wounds managed with enzymatic debridement using bromelain-based agent, NexoBrid®.
The antimicrobial properties of bromelain have been previously studied. However, the effect of enzymatic debridement on bacterial colonisation in burn wounds was not described in literature. In this study, we examine whether bromelain-based enzymatic debridement using NexoBrid® gel alters the microbiological pattern of burn wounds. Bacterial emergences in burn wounds that were enzymatically debrided at Pinderfields Regional Burns Centre, between July 2016 and February 2019, were studied and compared to the bacterial profile of burn wounds that were managed either by surgical debridement or dressings only during the same period. Our results showed that the microbial profile of burn wounds treated with NexoBrid® is similar to what is widely reported in cases treated without enzymatic debridement, at all stages of wound healing. This particularly showed in the predominance of Gram-positive organisms in the first week and Gram-negative in the second week.


Corneal shrinkage induced by nonmechanical Q-switched erbium:YAG laser trephination for penetrating keratoplasty in porcine eyes.
To assess the degree of corneal diameter shrinkage induced by Q-switched mid-infrared laser corneal trephination for penetrating keratoplasty in an experimental model. Corneal trephination was performed in 80 enucleated porcine eyes fixed in a holder centered on an automated globe rotation device, by Q-switched (2.94 microm) Er:YAG laser along open masks. Four types of masks were used to protect the underlying corneal tissue: metal masks (donor and recipient) and ceramic masks (donor and recipient). Two spot diameters (0.65 mm and 0.96 mm) were combined with two energy settings (40 mJ/pulse and 50 mJ/pulse) for each of the masks used. Repetition rate was fixed at 5 Hz. Diameters of donor buttons/recipient beds (horizontal and vertical) were measured immediately after the trephination and compared to the given mask size. Minimum corneal shrinkage was found in the recipient metal mask group (mean +/- SD=0.3+/-0.4%) with 50 mJ pulse energy and 0.65 mm spot diameter (in the horizontal diameter), while the maximum shrinkage (5.3+/-2.8%) was found in the donor metal mask group with 50 mJ pulse energy and 0.96 mm spot diameter. Corneal shrinkage was less pronounced in recipient beds than in donor buttons (P<0.01). The differences in shrinkage between the use of ceramic and metal masks were insignificant (P>0.05). Mean induced corneal diameter discrepancies between the donor button and the recipient bed (with metal and ceramic masks) were 2.5% and 2.5% in vertical diameter and 3.4% and 2.4% in horizontal diameter. The Q-switched Er:YAG laser experimental corneal trephination for penetrating keratoplasty may induce minor degrees of corneal diameter shrinkage in donor buttons and recipient openings. Oversizing of donor masks by 0.25-0.35 mm (i.e. 3-4% of graft size) may be a valid option to avoid refractive consequences.


Anaerobic alactic energy assessment in middle distance swimming.
To estimate the anaerobic alactic contribution in a 200 m middle distance swimming trial by means of two different methods based: (1) on the fast component of the VO2 off-kinetics (Ana recovery) and (2) on the kinetics of maximal phosphocreatine splitting in the contracting muscle (Ana pcr). Ten elite male swimmers performed a 200 m front crawl trial at maximal velocity during which VO2 was directly measured using a telemetric portable gas analyser; during the recovery period VO2 data were collected until baseline values were reached. No significant differences between the two methods were observed; mean ± SD values were 31.7 ± 2.5 and 32.6 ± 2.8 kJ, for Ana pcr and Ana recovery, respectively. Despite the existence of some caveats regarding both methods for estimation of the anaerobic alactic contribution, data reported in this study indicate that both yield similar results and both allow to estimate this contribution in supra-maximal swimming trials. This has important implications on swimming energetics, since the non-inclusion of the anaerobic alactic contribution to total metabolic energy expenditure leads to an underestimation of the energy cost at supra-maximal speeds.


Interatrial groove tear: an unusual complication of balloon atrial septostomy.
The authors present an unusual complication of the balloon atrial septostomy procedure performed in a neonate with D-transposition of the great arteries. Cardiac tamponade developed shortly after the balloon atrial septostomy procedure and the infant was found to have a tear in the superior aspect of the left atrium, parallel to the interatrial groove. The tear was successfully sutured and may have been caused by the greater pull-back force needed if the catheter balloon is maximally distended for the first pull-back.


Biased estimators of quantitative trait locus heritability and location in interval mapping.
In many empirical studies, it has been observed that genome scans yield biased estimates of heritability, as well as genetic effects. It is widely accepted that quantitative trait locus (QTL) mapping is a model selection procedure, and that the overestimation of genetic effects is the result of using the same data for model selection as estimation of parameters. There are two key steps in QTL modeling, each of which biases the estimation of genetic effects. First, test procedures are employed to select the regions of the genome for which there is significant evidence for the presence of QTL. Second, and most important for this demonstration, estimates of the genetic effects are reported only at the locations for which the evidence is maximal. We demonstrate that even when we know there is just one QTL present (ignoring the testing bias), and we use interval mapping to estimate its location and effect, the estimator of the effect will be biased. As evidence, we present results of simulations investigating the relative importance of the two sources of bias and the dependence of bias of heritability estimators on the true QTL heritability, sample size, and the length of the investigated part of the genome. Moreover, we present results of simulations demonstrating the skewness of the distribution of estimators of QTL locations and the resulting bias in estimation of location. We use computer simulations to investigate the dependence of this bias on the true QTL location, heritability, and the sample size.


Superwarfarin poisoning.
Superwarfarins consist of two classes of compounds, the 4-hydroxycoumarins and the indandiones. These compounds have replaced warfarin as an anticoagulant rodenticide. This article is a summary of the clinical effects produced by accidental and deliberate ingestions of superwarfarins. Most accidental ingestions occur in childhood and result in a benign outcome. Deliberate overdoses usually involve repeated ingestions of large quantities of superwarfarins that have resulted in prolonged laboratory evidence of interference with clotting; serious bleeding has been rare. This article suggests an approach to the management of patients with accidental and deliberate anticoagulant rodenticide ingestions.


Cavernous angioma of the optic nerve. Case report.
The case of a 24-year-old woman with a cavernoma of the right optic nerve is presented. She suffered recurrent headaches and showed a deficit of the right nasal visual field. A computed tomography scan, a computed tomography cisternography, and magnetic resonance imaging revealed a lesion in projection on the right suprasellar cistern, yet angiography was inconspicuous. On operation, an angiomatous tumor of the right optic nerve with a surrounding hemorrhage was found. Histological examination of the specimen confirmed the intraoperative diagnosis of a cavernoma. Postoperatively, the right nasal visual field deficit increased slightly.


More educated emergency department patients are less likely to receive opioids for acute pain.
Inadequate treatment of pain in United States emergency departments (EDs) is common, in part because of the limited and idiosyncratic use of opioids by emergency providers. This study sought to determine the relationship between patient socioeconomic characteristics and the likelihood that they would receive opioids during a pain-related ED visit. We conducted a cross-sectional analysis of ED data obtained as part of a multicenter study of outcomes after minor motor vehicle collision (MVC). Study patients were non-Hispanic white patients between the ages of 18 and 65 years who were evaluated and discharged home from 1 of 8 EDs in 4 states. Socioeconomic characteristics include educational attainment and income. Of 690 enrolled patients, the majority had moderate or severe pain (80%). Patients with higher education attainment had lower levels of pain, pain catastrophizing, perceived life threat, and distress. More educated patients were also less likely to receive opioids during their ED visit. Opioids were given to 54% of patients who did not complete high school vs 10% of patients with post-college education (χ(2) test P<.001). Differences in the frequency of opioid administration between patients with the lowest educational attainment (39%, 95% confidence interval 22% to 60%) and highest educational attainment (13%, 95% confidence interval 7% to 23%) remained after adjustment for age, sex, income, and pain severity (P=.01). In this sample of post-MVC ED patients, more educated patients were less likely to receive opioids. Further study is needed to assess the generalizability of these findings and to determine the reason for the difference.


A review of thresholding strategies applied to human chromosome segmentation.
Karyotype analysis is a widespread procedure in cytogenetics to assess the presence of genetic defects by the visualization of the structure of chromosomes. The procedure is lengthy and repetitive and an effective automatic analysis would greatly help the cytogeneticist routine work. Still, automatic segmentation and the full disentangling of chromosomes are open issues. The first step in every automatic procedure is the thresholding step, which detect blobs that represent either single chromosomes or clusters of chromosomes. The better the thresholding step, the easier is the subsequent disentanglement of chromosome clusters into single entities. We implemented eleven thresholding methods, i.e. the ones that appear in the literature as the best performers, and compared their performance in segmenting chromosomes and chromosome clusters in cytogenetic Q-band images. The images are affected by the presence of hyper- or hypo-fluorescent regions and by a contrast variability between the stained chromosomes and the background. A thorough analysis of the results highlights that, although every single algorithm shows peculiar strong/weak points, Adaptive Threshold and Region Based Level Set have the overall best performance. In order to provide the scientific community with a public dataset, the data and manual segmentation used in this paper are available for public download at http://bioimlab.dei.unipd.it.


Intercellular trafficking via plasmodesmata: molecular layers of complexity.
Plasmodesmata are intercellular pores connecting together most plant cells. These structures consist of a central constricted form of the endoplasmic reticulum, encircled by some cytoplasmic space, in turn delimited by the plasma membrane, itself ultimately surrounded by the cell wall. The presence and structure of plasmodesmata create multiple routes for intercellular trafficking of a large spectrum of molecules (encompassing RNAs, proteins, hormones and metabolites) and also enable local signalling events. Movement across plasmodesmata is finely controlled in order to balance processes requiring communication with those necessitating symplastic isolation. Here, we describe the identities and roles of the molecular components (specific sets of lipids, proteins and wall polysaccharides) that shape and define plasmodesmata structural and functional domains. We highlight the extensive and dynamic interactions that exist between the plasma/endoplasmic reticulum membranes, cytoplasm and cell wall domains, binding them together to effectively define plasmodesmata shapes and purposes.


New synthetic chitosan hybrids bearing some heterocyclic moieties with potential activity as anticancer and apoptosis inducers.
Wastes of the freshwater crayfish Procambarus clarkii were used as precursor for the extraction of chitin, which was deacetylation with alkali giving chitosan. Chitosan was transformed into the nanoparticles using sodium tripolyphosphate. Chitosan Cs and its nanoparticles CNP were characterized through FTIR, 1H NMR and surface morphology. Moreover, it was converted into some chitosan Schiff bases; CSB-1,2,3, containing different five membered heterocyclic moieties and also they were characterized. All of the synthesized compounds were evaluated against three cell lines, such as HepG-2, HCT-116 and MCF-7, whereas compounds CSB-2 and CSB-3 showed the highest cytotoxic activities. The higher activities for the latter compounds were confirmed through apoptosis studies (Caspase-3 and Bax/Bcl-2 ratio) against HePG-2 cells. Cell cycle analysis and apoptosis induction showed that compound CSB-2 induces apoptosis at pre G1 phase and cell growth arrest at G2/M phase.


Plant DNA Methyltransferase Genes: Multiplicity, Expression, Methylation Patterns.
Expression and methylation patterns of genes encoding DNA methyltransferases and their functionally related proteins were studied in organs of Arabidopsis thaliana plants. Genes coding for the major maintenance-type DNA methyltransferases, MET1 and CMT3, and the major de novo-type DNA methyltransferase, DRM2, are actively expressed in all organs. Similar constitutively active expression was observed for genes encoding their functionally related proteins, a histone H3K9 methyltransferase KYP and a catalytically non-active protein DRM3. Expression of the MET1 and CMT3 genes is significantly lower in developing endosperm compared with embryo. Vice versa, expression of the MET2a, MET2b, MET3, and CMT2 genes in endosperm is much more active compared with embryo. A special maintenance DNA methylation system seems to operate in endosperm. The DNMT2 and N6AMT genes encoding putative methyltransferases are constitutively expressed at low levels. CMT1 and DRM1 genes are expressed rather weakly in all investigated organs. Most of the studied genes have methylation patterns conforming to the "body-methylated gene" prototype. A peculiar feature of the MET family genes is methylation at all three possible site types (CG, CHG, and CHH). The most weakly expressed among genes of their respective families, CMT1 and DRM1, are practically unmethylated. The MET3 and N6AMT genes have unusual methylation patterns, promoter region, and most of the gene body devoid of any methylation, and the 3'-end proximal part of the gene body is highly methylated.


Correction to: Regulation of GAD65 expression by SMAR1 and p53 upon Streptozotocin treatment.
An amendment to this paper has been published and can be accessed via the original article.


Antibody ligation of murine Ly-6G induces neutropenia, blood flow cessation, and death via complement-dependent and independent mechanisms.
Ly-6G is a member of the Ly-6 family of GPI-linked proteins, which is expressed on murine neutrophils. Antibodies against Ly-6G cause neutropenia, and fatal reactions also develop if mice are primed with TNF-alpha prior to antibody treatment. We have investigated the mechanisms behind these responses to Ly-6G ligation in the belief that similar mechanisms may be involved in neutropenia and respiratory disorders associated with alloantibody ligation of the related Ly-6 family member, NB1, in humans. Neutrophil adhesion, microvascular obstruction, breathing difficulties, and death initiated by anti-Ly-6G antibodies in TNF-alpha-primed mice were shown to be highly complement-dependent, partly mediated by CD11b, CD18, and FcgammaR and associated with clustering of Ly-6G. Neutrophil depletion, on the other hand, was only partly complement-dependent and was not altered by blockade of CD11b, CD18, or FcgammaR. Unlike other neutrophil-activating agents, Ly-6G ligation did not induce neutropenia via sequestration in the lungs. Cross-linking Ly-6G mimicked the responses seen with whole antibody in vivo and also activated murine neutrophils in vitro. Although this suggests that the responses are, in part, mediated by nonspecific properties of antibody ligation, neutrophil depletion requires an additional mechanism possibly specific to the natural function of Ly-6G.


Smoking and cardiac symptoms as predictors of lung cancer.
In 1962, a cohort of 4604 Finnish men was interviewed about their smoking habits and cardiorespiratory symptoms. The cohort was followed up for deaths and incidence of lung cancer from 1963 to 1980 in order to study the effect of smoking and cardiac symptoms on the incidence of lung cancer. When analyzed simultaneously with smoking, the symptoms of angina, possible infarction and shortness of breath were all significantly associated with increased lung cancer risk. For example, the RR of lung cancer among those with possible infarction was 2.4, when age and smoking were adjusted for, and 1.8, when additionally shortness of breath and angina-like chest pain were adjusted for. Among smokers of greater than or equal to 15 g/day, those with symptoms of angina displayed a considerable excess risk (RR 2.5). A broad range of impairments of the cardiopulmonary functions seem to be associated with the carcinogenic processes invoked by smoking.


Anti-TNP antibody localization of the reactive lysine residues in myosin.
Myosin contains reactive lysine residues which are trinitrophenylated by 2,4,6-trinitrobenzene sulfonate much faster than the rest of the lysines. Here we find the location of these residues in the primary and spatial structure of myosin with the help of an anti-trinitrophenyl antibody. This antibody was raised against trinitrophenyl hemocyanin in rabbits. It reacted with trinitrophenylated myosin, and with some of the tryptic fragments of trinitrophenylated myosin. By analyzing the reaction with Western blots, it was found that the antibody preferentially reacts with the 27 kDa N-terminal fragment of the myosin head, and more weakly with the light meromyosin region of the myosin rod. The 27 kDa fragment contains the most reactive lysine residue, while the intermediate lysine residue is located in the light meromyosin region. The locations of the epitopes of the antibody were visualized on electron microscope images of rotary-shadowed trinitrophenylated myosin-antibody complexes. The distances of the epitopes to the head-rod junction of myosin were measured as 13 and 113 nm for the epitope on the head (reactive lysine residue) and for that on the rod (intermediary reactive lysine residue), respectively.