Root Conditioning with Citric Acid and Ethylenediaminetetraacetic Acid and their Effect on Fibrin Clot Adhesion to Dentin-A Scanning Electron Microscopic Study.
Root biomodifiers are used to remove the smear layer formed on the roots as a result of mechanical debridement and also to expose collagen fibers making the root surfaces biologically acceptable. Further, the blood element linkage to demineralized roots and clot stabilization by collagen fibres are vital for the wound healing and success of periodontal surgery. The aim of this study was to evaluate and compare the distribution of fibrin network to dentin surfaces conditioned with citric acid and Ethylenediaminetetraacetic Acid (EDTA) using Scanning Electron Microscope (SEM). Thirty human dentin blocks (4 mm x 3 mm x 1 mm) were prepared and equally divided into three groups. The control Group A specimens were treated with Phosphate Buffered Saline (PBS). The study Group B and C specimens were treated with citric Acid and EDTA respectively. After conditioning, a drop of fresh human whole blood was placed on the dentin specimens and was allowed to clot. The specimens were then processed using standardized protocol for SEM. The dentin surfaces were then analysed for fibrin network linkage under SEM and scored according to the degree of fibrin linkage to the dentin surface. Statistical analysis was done using Kruskal Wallis test to compare the score values between the study groups. Pairwise comparison between the study groups was done using Mann Whitney U test with Bonferroni correction. The fibrin network formed on the dentin network were analysed and scores were given. Kruskal Wallis test and Mann Whitney U test with Bonferroni correction were used for statistical analysis. It was found that most of the dentin specimens conditioned with citric acid and covered with human blood exhibited dense fibrin network compared to EDTA. However, statistical analysis showed that there were no significant differences found between the groups (p=0.35). Citric acid and EDTA has almost comparable effect in promoting the adhesion of fibrin network onto the dentin surface which may further enhance connective tissue attachment to the root surface.


Oblique orientation discrimination thresholds are superior in those with a high level of autistic traits.
Enhanced low-level perception, although present in individuals with autism, is not seen in individuals with high, but non-clinical, levels of autistic traits (Brock et al.in Percept Lond 40(6):739. doi: 10.1068/p6953 , 2011). This is surprising, as many of the higher-level visual differences found in autism have been shown to correlate with autistic traits in non-clinical samples. Here we measure vertical-oblique and, more difficult, oblique-oblique orientation discrimination thresholds in a non-clinical sample. As predicted, oblique-oblique thresholds provided a more sensitive test of orientation discrimination, and were negatively related to autistic traits (N = 94, r = -.356, p < .0001). We conclude that individual differences in orientation discrimination and autistic traits are related, and suggest that both of these factors could be mediated by increased levels of the inhibitory neurotransmitter GABA.


Antioxidant Activity of Egg Yolk Protein Hydrolysates Obtained by Enzymatic and Sub-Critical Water Hydrolysis.
Obtaining peptides with antioxidant properties by enzymatic hydrolysis has been widely described; however, the use of non-enzymatic methods to obtain peptides with antioxidant capacities has been poorly investigated. In this study, non-soluble proteins obtained from delipidated egg yolk granules were hydrolyzed with trypsin, and with a non-enzymatic method using sub-critical water hydrolysis under a non-oxidizing (nitrogen) and oxidizing (oxygen) atmosphere. The effect of the sub-critical water hydrolysis on the amino acids' composition of the hydrolysates was assessed. Furthermore, the antioxidant capacities of the hydrolysates were evaluated using the ABTS•+ scavenging assay, the DPPH radical scavenging activity assay, and by measuring the reducing power of the peptides, the peptides' ferrous ion chelating capacities, and the antioxidant effect of the peptides on beef homogenates. The hydrolysate obtained by sub-critical water hydrolysis under a nitrogen stream showed similar or better results in the antioxidant tests than those obtained using trypsin hydrolysis, except in the ferrous chelating capacity, where the trypsin hydrolysate showed the best performance. The oxidizing environment promoted by the oxygen in the other sub-critical water hydrolysis method tested produced the peptides with the lowest antioxidant capacities, due to changes in the primary structure of the peptides. These results suggest that the sub-critical water hydrolysis method under a nitrogen stream, in comparison with the enzymatic hydrolysis, is a reliable method to obtain peptides with good antioxidant capacities.


Comparison of the effects of standard vs low-dose prolonged-release tacrolimus with or without ACEi/ARB on the histology and function of renal allografts.
Targeting the renin-angiotensin system and optimizing tacrolimus exposure are both postulated to improve outcomes in renal transplant recipients (RTRs) by preventing interstitial fibrosis/tubular atrophy (IF/TA). In this multicenter, prospective, open-label controlled trial, adult de novo RTRs were randomized in a 2 × 2 design to low- vs standard-dose (LOW vs STD) prolonged-release tacrolimus and to angiotensin-converting enzyme inhibitors/angiotensin II receptor 1 blockers (ACEi/ARBs) vs other antihypertensive therapy (OAHT). There were 2 coprimary endpoints: the prevalence of IF/TA at month 6 and at month 24. IF/TA prevalence was similar for LOW vs STD tacrolimus at month 6 (36.8% vs 39.5%; P = .80) and ACEi/ARBs vs OAHT at month 24 (54.8% vs 58.2%; P = .33). IF/TA progression decreased significantly with LOW vs STD tacrolimus at month 24 (mean [SD] change, +0.42 [1.477] vs +1.10 [1.577]; P = .0039). Across the 4 treatment groups, LOW + ACEi/ARB patients exhibited the lowest mean IF/TA change and, compared with LOW + OAHT patients, experienced significantly delayed time to first T cell-mediated rejection. Renal function was stable from month 1 to month 24 in all treatment groups. No unexpected safety findings were detected. Coupled with LOW tacrolimus dosing, ACEi/ARBs appear to reduce IF/TA progression and delay rejection relative to reduced tacrolimus exposure without renin-angiotensin system blockade. ClinicalTrials.gov identifier: NCT00933231.


Thermally Stable Low-Loss Polymer Dielectrics Enabled by Attaching Cross-Linkable Antioxidant to Polypropylene.
Polymer dielectrics with low-loss and high-temperature tolerance are extremely desirable as electrical energy storage materials for advanced electronics and electrical power applications. They can allow fast switching rates during power conversion and therefore achieve high power densities without thermal issues. Here, we explore polypropylene (PP), the state of the art dielectric polymer, and present an innovative approach to substantially improve the thermal stability and concurrently reduce the dielectric loss of PP. In particular, cross-linkable antioxidant groups, hindered phenol (HP), are incorporated into PP via well-controlled chemical synthesis. The grafted HP can simultaneously serve as radical scavenger and cross-linker, thereby constraining thermally decomposed radicals and charge transport in the synthesized PP-HP copolymer. As a result, the upper-temperature limit of PP-HP is greatly extended to 190 °C and the electrical loss is even gradually reduced upon thermal annealing. The copolymer after heating under 190 °C exhibits better dielectric properties than the PP without any thermal treatment. The experimental results indicate that the PP-HP copolymers are promising materials for high-temperature, low-loss, and high-voltage dielectric applications.


Brazilian Portuguese translation, cross-cultural adaptation, and content validation of the pediatric lower urinary tract symptom score.
Early screening instruments for professionals are important to reduce the cost and time of diagnosing pediatric lower urinary tract dysfunction. This study aimed to translate, cross-culturally adapt, and validate pediatric lower urinary tract symptom scores. This methodological study was conducted in 2019 in two phases: translation and cross-cultural adaptation, and content validation. In the first phase, we used translators, specialists, and 30 parents of children with voiding dysfunction. In the second phase, validation was performed by 11 pediatric urology specialists. The instrument was translated and adapted to Brazilian Portuguese and presented an excellent level of verbal comprehension. Further, the instrument presented a good content validity index >0.8 for all items, except for the nomenclature and clarity of two items, as indicated by the Cronbach's alpha value of 0.75 for internal consistency. The final version showed semantic, idiomatic, experiential, and conceptual equivalence with the original instrument, indicating that it is adequate for other assessments of psychometric qualities. The translation and adaptation of an important instrument for diagnosing lower urinary tract dysfunction in children to other countries helps nurses identify the disease early and monitor the treatment.


The mitochondrial-targeted compound SS-31 re-energizes ischemic mitochondria by interacting with cardiolipin.
Ischemia causes AKI as a result of ATP depletion, and rapid recovery of ATP on reperfusion is important to minimize tissue damage. ATP recovery is often delayed, however, because ischemia destroys the mitochondrial cristae membranes required for mitochondrial ATP synthesis. The mitochondria-targeted compound SS-31 accelerates ATP recovery after ischemia and reduces AKI, but its mechanism of action remains unclear. Here, we used a polarity-sensitive fluorescent analog of SS-31 to demonstrate that SS-31 binds with high affinity to cardiolipin, an anionic phospholipid expressed on the inner mitochondrial membrane that is required for cristae formation. In addition, the SS-31/cardiolipin complex inhibited cytochrome c peroxidase activity, which catalyzes cardiolipin peroxidation and results in mitochondrial damage during ischemia, by protecting its heme iron. Pretreatment of rats with SS-31 protected cristae membranes during renal ischemia and prevented mitochondrial swelling. Prompt recovery of ATP on reperfusion led to rapid repair of ATP-dependent processes, such as restoration of the actin cytoskeleton and cell polarity. Rapid recovery of ATP also inhibited apoptosis, protected tubular barrier function, and mitigated renal dysfunction. In conclusion, SS-31, which is currently in clinical trials for ischemia-reperfusion injury, protects mitochondrial cristae by interacting with cardiolipin on the inner mitochondrial membrane.


Ligand-triggered resistance to molecular targeted drugs in lung cancer: roles of hepatocyte growth factor and epidermal growth factor receptor ligands.
Recent advances in molecular biology have led to the identification of new molecular targets, such as epidermal growth factor receptor ( EGFR ) mutations and echinoderm microtubule-associated protein-like 4 (EML4) - anaplastic lymphoma kinase (ALK) fusion gene, in lung cancer. Dramatic response has been achieved with EGFR inhibitors (gefitinib and erlotinib) and an ALK inhibitor (crizotinib) in lung cancer expressing corresponding targets. However, cancer cells acquire resistance to these drugs and cause recurrence. Known major mechanisms for resistance to molecular targeted drugs include gatekeeper mutations in the target gene and activation of bypass survival signal via receptors other than the target receptors. The latter mechanism can involve receptor gene amplification and ligand-triggered receptor activation as well. For example, hepatocyte growth factor (HGF), the ligand of a tyrosine kinase receptor Met, activates Met and the downstream PI3K/Akt pathway and triggers resistance to EGFR inhibitors in EGFR mutant lung cancer cells. Moreover, EGFR ligands activate EGFR and downstream pathways and trigger resistance to crizotinib in EML4-ALK lung cancer cells. These observations indicate that signals from oncogenic drivers (EGFR signaling in EGFR -mutant lung cancer and ALK signaling in EML4-ALK lung cancer) and ligand-triggered bypass signals (HGF-Met and EGFR ligands-EGFR, respectively) must be simultaneously blocked to avoid the resistance. This review focuses specifically on receptor activation by ligand stimulation and discusses novel therapeutic strategies that are under development for overcoming resistance to molecular targeted drugs in lung cancer.


Efficacy and tolerability of a new synergized pyrethrins thermofobic foam in comparison with benzyl benzoate in the treatment of scabies in convicts: the ISAC study (Studio Della scabbia in ambiente carcerario).
Scabies is a very common skin infection in convicts. The SIMSPE Society (Società Italiana di Medicina e Sanità Penitenziaria) has organized and conducted a multicentre, randomized, comparative, parallel group, investigator-blinded trial to evaluate the efficacy and tolerability of synergized pyrethrins foam (PF) in comparison with benzyl benzoate (BB) lotion. A total of 240 convicted patients, enrolled in eight National Jail Institutions, with a clinical diagnosis of scabies, were treated with PF (n = 120) for three consecutive days or BB (n = 120) for five consecutive days. Primary study endpoints were the clinical cure rate and the local tolerability. Secondary endpoints were clinical evolution of scabietic lesions and itching intensity. Study outcomes were assessed using appropriate semiquantitative scores at baseline and after 2 and 4 weeks. A second treatment cycle was applied if after 2 weeks the patient was not judged clinically cured. At week 2, a total of 75% (95% CI: 66-82%) and 71% (95% CI: 62-78%) of patients showed a complete clinical cure rate in the PF and BB groups, respectively. At week 4, the percentage of totally cured patients increased up to 95% (95% CI: 89-97%) and 91% (95% CI: 83-94%) in the PF and BB groups, respectively (P = NS between groups). At week 4, 5% in the PF group and 9% in the BB group complained of itching. Burning and irritation after treatment applications were more common in the BB group in comparison with the PF group. The tolerability score was better in the PF group in comparison with to BB group (2.9 vs. 2.2; P = 0.0001). A total of 95% of patients in the PV group had a good tolerability score (i.e. = 3) in comparison with 41% in the BB group. Our results show that a 3-day treatment with pyrethrins thermofobic foam is at least as effective as a 5-day treatment with benzyl benzoate lotion in convicted subjects with scabies. The foam formulation is better tolerated than the benzyl benzoate lotion.


Ultraviolet mutagenesis and its repair in an Escherichia coli strain containing a nonsense codon.
Ultraviolet mutagenesis and its repair were studied mainly in WU36-10-89, a uvr(-) strain of Escherichia coli containing a UAG mutation in a gene for leucine biosynthesis. Following ultraviolet (UV) irradiation revertants appearing with or without direct photoreactivation (PR) were classified according to the presence and type of suppressor they contained. We find UV mutation production to be quite specific. An analysis of revertants produced by UV indicates they are formed mainly from GC --> AT and that the miscoding is due to a cytosine residue at the site of mutation in a cytosine-thymine (CT) dimer. We propose that the dimer serves as template during some aspects of repair replication and at the time of replication the C in the dimer directs the insertion of A in the complementary strand. We also note that C --> A and T -->G changes caused by a CT dimer occur much less frequently.


The distribution of the outer gas vesicle protein, GvpC, on the Anabaena gas vesicle, and its ratio to GvpA.
Previous studies have shown that gas vesicles isolated from the cyanobacterium Anabaena flos-aquae contain two types of protein, GvpA, a small hydrophobic protein that forms the main ribbed structure, and GvpC, a protein comprising five repeats of a 33-amino-acid-residue motif, which is located on the outer surface of the GvpA shell. GvpC was shown to increase the critical collapse pressure of the gas vesicles; it was thought to do this by forming a series of molecular ties that bind the ribs together. We now show that antibodies raised against GvpC label both the central cylinders and the conical end caps of native gas vesicles but fail to bind to gas vesicles that have been stripped of GvpC. The molar ratio of GvpA to GvpC has been calculated from amino acid analyses of gas vesicle hydrolysates by reference to the abundance of amino acids that occur predominantly or exclusively in one protein or the other; the molar ratio was found to be 25:1 in freshly isolated gas vesicles and 23:1 in gas vesicles saturated with GvpC. We have considered three ways in which the 33-residue repeats of GvpC might interact with the crystallographic unit cell of GvpA molecules in the ribs. The Anabaena GvpC will bind to and restore the strength of gas vesicles isolated from Aphanizomenon and Microcystis that lack their native GvpC.


Editorial: just how "difficult" is it to withdraw PPI treatment?
Two recent studies-one of which is published in this edition of the American Journal of Gastroenterology-have reported that new dyspeptic or reflux symptoms may develop among previously asymptomatic individuals after withdrawal of a short course of a proton pump inhibitor (PPI). It has been suggested that this is attributable to rebound gastric acid hypersecretion, and that the same phenomenon explains why some patients become chronic PPI users and physicians experience "difficulty" in withdrawing PPI treatment from patients. Although we congratulate the authors of both studies for their controlled observations, we question the generalizability of these observations and hypotheses to the clinical domain. PPI treatment continues to be the optimal management strategy for most patients with gastroesophageal reflux disease and is indicated for chronic use as ulcer prophylaxis in nonsteroidal anti-inflammatory drug takers at high risk for bleeding. However, as with all drugs, PPIs should be dosed appropriately, and should be reserved for patients with conditions for which there is clear evidence of benefit from therapy.


Disruption of inducible 6-phosphofructo-2-kinase impairs the suppressive effect of PPARγ activation on diet-induced intestine inflammatory response.
PFKFB3 is a target gene of peroxisome proliferator-activated receptor gamma (PPARγ) and encodes for inducible 6-phosphofructo-2-kinase (iPFK2). As a key regulatory enzyme that stimulates glycolysis, PFKFB3/iPFK2 links adipocyte metabolic and inflammatory responses. Additionally, PFKFB3/iPFK2 is involved in the effect of active PPARγ on suppressing overnutrition-induced adipose tissue inflammatory response, which accounts for the insulin-sensitizing and antidiabetic effects of PPARγ activation. Using PFKFB3/iPFK2-disrupted mice, the present study investigated the role of PFKFB3/iPFK2 in regulating overnutrition-associated intestine inflammatory response and in mediating the effects of PPARγ activation. In wild-type mice, intestine PFKFB3/iPFK2 was increased in response to high-fat diet (HFD) feeding compared with that in mice fed a low-fat diet. However, intestine PFKFB3/iPFK2 was decreased in PFKFB3/iPFK2-disrupted mice and did not respond to HFD feeding. Furthermore, on an HFD, PFKFB3/iPFK2-disrupted mice displayed a significant increase in major intestine proinflammatory indicators such as toll-like receptor 4 expression, c-Jun N-terminal kinase 1 and nuclear factor kappa B phosphorylation, and proinflammatory cytokine expression compared with wild-type littermates. Upon treatment with rosiglitazone, an agonist of PPARγ, intestine proinflammatory indicators were markedly decreased in wild-type mice, but to a much lesser degree in PFKFB3/iPFK2-disrupted mice. Overall, the status of HFD-induced intestine inflammatory response in all treated mice correlated inversely with systemic insulin sensitivity, indicated by the homeostasis model assessment of insulin resistance data. Together, these results suggest that PFKFB3/iPFK2 is critically involved in the effect of PPARγ activation on suppressing diet-induced intestine inflammatory response.


Survival after surgical management of pancreatic adenocarcinoma: does curative and radical surgery truly exist?
Surgery for pancreatic cancer offers a low success rate but it provides the only likelihood of cure. Modern series show that, in experienced hands, the standard Whipple procedure is associated with a 5-year survival of 10%-20%, with a perioperative mortality rate of less than 5%. Most patients, however, will develop recurrent disease within 2 years after curative treatment. This occurs, usually, either at the site of resection or in the liver. This suggests the presence of micrometastases at the time of operation. Negative lymph nodes are the strongest predictor for long-term survival. Other predictors for a favourable outcome are tumour size, radical surgery and a histopathologically well-differentiated tumour. Adjuvant therapy has, so far, shown only modest results, with 5FU chemotherapy, to date, the only proven agent able to increase survival. Nowadays, the choice of therapy should be based on histopathological assessment of the tumour. Knowledge of the molecular basis of pancreatic cancer has led to various discoveries concerning its character and type. Well-known examples of genetic mutations in adenocarcinoma of the pancreas are k-ras, p53, p16, DPC4. Use of molecular diagnostics and markers in the assessment of tumour biology may, in future, reveal important subtypes of this type of tumour and may possibly predict the response to adjuvant therapy. Defining the subtypes of pancreatic cancer will, hopefully, lead to target-specific, less toxic and finally more effective therapies. Long-term survival is observed in only a very small group of patients, contradicting the published actuarial survival rates of 10%-45%. Assessment of clinical benefit from surgery and adjuvant therapy should, therefore, not only be based on actuarial survival but also on progression-free survival, actual survival, median survival and quality of life (QOL) indicators. Survival in surgical series is usually calculated by actuarial methods. If there is no information on the total number of patients and the number of actual survivors, and no clear definition of the subset of patients, actuarial survival curves can prove to be misleading. Proper assessment of QOL after surgery and adjuvant therapy is of the utmost importance, as improvements in survival rates have, so far, proved to be disappointing.


Adipose tissue content as a modifier of the tissue distribution, biological effects, and excretion of a hexachlorobiphenyl in C57BL/6J and DBA/JBOMf mice.
C57BL/6J (C57) and DBA/JBOMf (DBA) mice were used to study the role of adipose tissue as a modifier of tissue distribution, biological effects, and elimination of a lipophilic foreign chemical, 2,4,5,2',4',5'-hexachlorobiphenyl (HCB). As an indication of biological potency of the model compound, the activities of hepatic drug-metabolizing enzymes were determined. DBA mice contained twice as much body fat as C57 mice. Since the highly lipophilic HCB was primarily sequestered by the adipose tissue, DBA mice required greater doses of HCB than did C57 mice to reach similar tissue levels of the chemical. Accordingly, greater HCB doses were required by DBA mice for elevation of drug-metabolizing enzyme activities. Phenobarbital elevated enzyme activities in a similar way in both mouse strains. When the dietary intake of DBA mice was restricted, the body fat content decreased from 15% to 5% of body weight during 1 week. In these animals the tissue accumulation of HCB and enzyme induction resembled the situation in C57 mice fed ad libitum. Highest elevations were seen in the activities of 7-ethoxycoumarin-O-deethylase and arylhydrocarbon hydroxylase (EC 220.127.116.11). In addition, the activity of epoxide hydrolase (EC 126.96.36.199) was increased, whereas glutathione S-transferase as well as UDP-glucuronosyltransferase (EC 126.96.36.199) activities remained unchanged. The abundant adipose tissue content played no role in the nonresponsiveness of DBA mice to 3-methylcholanthrene since, in contrast to C57 mice, no changes in enzyme activities were detected in DBA mice deprived of food, even after large doses of 3-methylcholanthrene. The adipose tissue content also affected the rate of elimination of HCB. DBA mice excreted smaller quantities of HCB than did C57 mice after equal doses. When, however, fasted DBA mice received HCB, they excreted it at rates similar to those of C57 mice fed ad libitum. In C57 mice, concomitant to the elevation of monooxygenase activities, there was an increase in the rate of excretion of HCB. No such elevation could be seen after a dose that was too small to elevate enzyme activities.


Higher risk of gastrointestinal parasite infection at lower elevation suggests possible constraints in the distributional niche of Alpine marmots.
Alpine marmots Marmota marmota occupy a narrow altitudinal niche within high elevation alpine environments. For animals living at such high elevations where resources are limited, parasitism represents a potential major cost in life history. Using occupancy models, we tested if marmots living at higher elevation have a reduced risk of being infected with gastro-intestinal helminths, possibly compensating the lower availability of resources (shorter feeding season, longer snow cover and lower temperature) than marmots inhabiting lower elevations. Detection probability of eggs and oncospheres of two gastro-intestinal helminthic parasites, Ascaris laevis and Ctenotaenia marmotae, sampled in marmot feces, was used as a proxy of parasite abundance. As predicted, the models showed a negative relationship between elevation and parasite detectability (i.e. abundance) for both species, while there appeared to be a negative effect of solar radiance only for C. marmotae. Site-occupancy models are used here for the first time to model the constrains of gastrointestinal parasitism on a wild species and the relationship existing between endoparasites and environmental factors in a population of free-living animals. The results of this study suggest the future use of site-occupancy models as a viable tool to account for parasite imperfect detection in eco-parasitological studies, and give useful insights to further investigate the hypothesis of the contribution of parasite infection in constraining the altitudinal niche of Alpine marmots.


Thirty-five cases of transient hyperthyroidism.
Over a 7-year period transient hyperthyroidism was diagnosed in 35 patients seen in a consulting practice in a community hospital. The patients were followed up for an average of 15 months. Initially all of them had biochemical evidence of hyperthyroidism but a very low 24-hour uptake of radioiodine. The hyperthyroid phase was short, and there were no relapses. Seventeen patients subsequently became hypothyroid; this phase, too, was almost always transient. The clinical course of the disease in the 11 women who became hyperthyroid within 6 months after giving birth was similar to that experienced by the other patients, but of the 11 who had increased titres of antimicrosomal antibodies a significantly greater proportion (73%) showed at least transient evidence of hypothyroidism; 1 patient remained frankly hypothyroid for a year. Transient hyperthyroidism can be distinguished from Graves' disease only if the uptake of radioiodine is measured. It is important to make this distinction, as transient hyperthyroidism can be managed safely and symptomatically with beta-blockers alone. The propensity of this disease for the postpartum period and the high proportion of patients with antithyroid antibodies suggest an autoimmune cause.


The origin and evolution of ribonucleotide reduction.
Ribonucleotide reduction is the only pathway for de novo synthesis of deoxyribonucleotides in extant organisms. This chemically demanding reaction, which proceeds via a carbon-centered free radical, is catalyzed by ribonucleotide reductase (RNR). The mechanism has been deemed unlikely to be catalyzed by a ribozyme, creating an enigma regarding how the building blocks for DNA were synthesized at the transition from RNA- to DNA-encoded genomes. While it is entirely possible that a different pathway was later replaced with the modern mechanism, here we explore the evolutionary and biochemical limits for an origin of the mechanism in the RNA + protein world and suggest a model for a prototypical ribonucleotide reductase (protoRNR). From the protoRNR evolved the ancestor to modern RNRs, the urRNR, which diversified into the modern three classes. Since the initial radical generation differs between the three modern classes, it is difficult to establish how it was generated in the urRNR. Here we suggest a model that is similar to the B12-dependent mechanism in modern class II RNRs.


Diazepam binding of dissociated hippocampal cultures from fetal mice.
Primary cultures of dissociated hippocampi from fetal mice examined for the presence of binding sites for [3H]diazepam. The binding assays were done with living cells still attached to the culture dish. The cells contain high affinity binding sites for [3H]diazepam, Kd = 5 nM, which are completely inhibited with 20 nM R05-4864 but only 26% with 20 nM lorazepam. The binding was inhibited by purinergic compounds and by quinidine. The living cell did not exhibit increased binding of [3H]diazepam in the presence of GABA and in fact a slight decrease in binding was found. This was also found when live, intact C6 glial cells were investigated. These observations suggest that the use of living cells to study the benzodiazepine receptor is valuable and maybe necessary to fully characterize this receptor.


Erectile dysfunction in a sample of patients attending a psychiatric outpatient department.
To assess the prevalence of erectile dysfunction among a sample of patients attending a psychiatric clinic, we administered the International Index of Erectile Function on 108 male patients attending the clinic. We also obtained sociodemographic and clinical variables from the patients. The mean age of the patients studied was 39.6 (s.d.=11.6), with a mean duration of illness of 10.24 (s.d.=8.2) years. There were 46.8% schizophrenics; other diagnoses include bipolar affective disorder, recurrent depressive disorder and substance use disorder. The prevalence of erectile dysfunction was 83%. Age, marital status, current medications and the presence of comorbid medical conditions were significantly associated with erectile dysfunction; however, only age and marital status significantly predicted erectile dysfunction. We concluded that erectile dysfunction is highly prevalent among patients attending the psychiatric clinic, as such inquiries about sexual function should be routinely carried out by clinicians.


Suppression of Gadd45 alleviates the G2/M blockage and the enhanced phosphorylation of p53 and p38 in zinc supplemented normal human bronchial epithelial cells.
An adequate zinc status is essential for optimal cellular functions and growth. Yet, excessive zinc supplementation can be cytotoxic and can impair cell growth. Gadd45 plays a vital role as cellular stress sensor in the modulation of cell signal transduction in response to stress. The present study was designed to determine the influence of zinc status on Gadd45 expression and cell cycle progression in zinc deficient and supplemented normal human bronchial epithelial (NHBE) cells, and to decipher the molecular mechanism(s) exerted by the suppression of Gadd45 expression on cell growth and cell cycle progression in this cell type. Cells were cultured for one passage in different concentration of zinc: <0.4 muM (ZD) as severe zinc deficient; 4 muM as normal zinc level in culture medium; 16 microM (ZA) as normal human plasma zinc level; and 32 muM (ZS) as the high end of plasma zinc attainable by oral supplementation. Inhibition of cell growth, upregulation of Gadd45 mRNA and protein expression, and blockage of G2/M cell cycle progression were observed in ZS cells. In contrast, little or no changes in these parameters were seen in ZD cells. The siRNA-mediated knocking down of Gadd45 was found to relieve G2/M blockage in ZS cells, which indicated that the blockage was Gadd45 dependent. Moreover, the enhanced phosphorylation of p38 and p53 (ser15) in ZS cells was normalized after suppression of Gadd45 by siRNA, implicating that the enhanced phosphorylation of these proteins was Gadd45 dependent. Thus, we demonstrated for the first time that an elevated zinc status modulated signal transduction to produce a delay at G2/M during cell cycle progression in NHBE cells.


Brachio-ankle pulse wave velocity and cardio-ankle vascular index (CAVI).
In order to diagnose arteriosclerosis in any part of the body, pulse wave velocity (PWV) measurement is a useful approach. However, it is considered that the technique of PWV measurement should be simplified. A new method for measuring PWV has therefore been proposed in Japan. The PWV of the brachial artery (ba) and the ankle was measured by applying air pressure with the aid of a volume plethysmograph. Comparisons between the baPWV measurement method and the conventional method are currently being performed. Since satisfactory results have been obtained to date, baPWV has gained popularity throughout Japan. Since this method measures PWV in the arm and foot, it may be said that aortic PWV is not reflected though a large amount of past PWV measurements. BaPWV is influenced by blood pressure. With the baPWV technique, blood pressure compensation is not carried out. Furthermore, the pulse pressure is measured by air pressure; therefore any stimulus that exerts pressure on an artery may influence these results. Due to these reasons, a cardie-ankle vascular index (CAVI) has been proposed in which the pressure wave form indicating the closing of the aortic valve appears in the form of an arterial pressure wave after a fixed delay time. This delay is the time difference between the actual closing of the aortic valve and the measuring point. Prior to the introduction of baPWV, PWV was measured in the carotid artery and foot. As in traditional PWV, baPWV uses the delay time, but between the brachial artery and the ankle artery. However, the carotid artery differs from the brachial artery, and the measured value differs depending on whether the arteriosclerosis is present in the carotid artery or the brachial artery. CAVI is calculated from the ECG, PCG, brachial artery waveform and ankle artery waveform using a special algorithm. This new method represents a breakthrough in the diagnosis of atherosclerosis.


Isavuconazole to prevent invasive fungal infection in immunocompromised adults: Initial experience at an academic medical centre.
To evaluate clinical and economic outcomes associated with the use of isavuconazole as antifungal prophylaxis in high-risk immunocompromised patients. Retrospective, single-centre cohort study of patients who received isavuconazole prophylaxis. Outcomes assessed included breakthrough IFI, early discontinuation of isavuconazole for any reason and antifungal prophylaxis prescribed at discharge. The impact on inpatient drug expenditure was evaluated using current isavuconazole and posaconazole drug costs per observed isavuconazole days of therapy (DOT) during the study period. One hundred thirty-eight courses of isavuconazole prophylaxis were administered to 98 inpatients (2193 DOT). Relapsed/refractory acute myelogenous leukaemia was the indication for prophylaxis in over half (59.4%) of patients. Breakthrough IFI occurred in 8 (5.8%) courses. Suspected drug-related toxicities led to early discontinuation in 6 (4.3%) courses (five hepatotoxicity, one drug rash). At discharge, 24 (17.4%) courses lacked insurance coverage for isavuconazole. The formulary switch to isavuconazole prophylaxis resulted in an estimated mean drug cost savings of $128.25 per DOT relative to estimated posaconazole costs (P < 0.001). Isavuconazole may be an option for antifungal prophylaxis in high-risk immunocompromised adults and has the potential to produce significant inpatient drug cost savings. Further studies are needed to confirm the clinical efficacy and cost-effectiveness of isavuconazole in this role.


Coexistence of amyloidosis and light chain deposition disease in the heart.
There are few reports on the coexistence of cardiac amyloid light-chain (AL) amyloidosis and light chain deposition disease (LCDD), despite their similar pathophysiologies caused by plasma-cell dyscrasia. Herein, we report the coexistence of these diseases. A 59-year-old man was referred to our hospital because of exertional dyspnea and hypotension. Renal dysfunction of unknown etiology had been present for 4 years and hemodialysis had been introduced. Severe systolic and diastolic cardiac dysfunction was apparent, accompanied with dilatation and granular sparkling, but not with left ventricular hypertrophy. The plasma-free light chain κ was found to be extremely high, with a κ/λ ratio of 1,919. Light microscopic examination of the endomyocardial biopsy revealed spotty and homogenous deposits, which positively stained with Congo red, and exhibited a blazing apple-green color under polarized light. Based on these results, cardiac amyloidosis was diagnosed. In specimens prepared for electron microscopy, no amyloid fibrils could be found. Instead, we observed amorphous nonfibrillar deposits around several small vessels including capillaries and small arteries, which were consistent with light-chain deposits. LCDD was diagnosed based on the systemic increase in κ light chain and the ultrastructural findings of the endomyocardial biopsy specimens. Coexistence of cardiac amyloidosis and LCDD was thus confirmed in our patient. An electron microscopic assessment in addition to Congo red staining may be useful to diagnose latent LCDD in patients with suspected cardiac light-chain amyloidosis.


Speech fluency in bilinguals who stutter: Language proficiency and attentional demands as mediating factors.
The current study examines how speech disfluencies manifest themselves in the two languages of bilingual persons who stutter, starting from the hypothesis that stuttering is associated with an attentional deficit at the level of speech production. Twenty-eight bilingual people who stutter performed a spontaneous and a controlled speech production task, once in their dominant and once in their non-dominant language. The controlled production task (i.e. a network description task) was carried out once under a full-attention condition and once under a divided-attention condition where a non-linguistic, pitch discrimination task was performed simultaneously. In both the spontaneous and the controlled speech task, bilingual persons who stutter produced more (typical and stuttering-like) disfluencies in their L2 than in their L1. Furthermore, whereas the typical disfluencies increased when attention was directed away from speech production, stuttering-like disfluencies decreased. This effect was however restricted to L2. In addition, L2 proficiency was generally found to be a predicting factor, with higher proficiency leading to fewer disfluencies. These results suggest that speaking in a non-dominant language increases both typical and stuttering-like disfluencies in bilingual persons who stutter, but also that these two types of dysfluencies differ regarding their attentional origins. Our findings offer further support for attentional accounts of stuttering and have both theoretical and clinical implications.


When inferiority meets non-inferiority: implications for interim analyses.
The objective of a non-inferiority trial is to determine whether a new or existing treatment is not less effective than another existing or current treatment by more than a pre-specified margin, Δ, usually with the requirement that the new treatment has some other advantage such as reduced cost or lower toxicity. A possible but unusual result in a non-inferiority trial is for the confidence interval for the treatment effect to lie between zero and Δ, implying that the new treatment is both inferior and non-inferior to the control. Such a result could occur in non-inferiority trials with large sample sizes or large non-inferiority margins. The possibility of this scenario occurring has implications for interim analyses. In standard superiority trials, stopping guidelines are often based on the p value obtained from testing whether treatments are equally effective. In non-inferiority trials, however, even if a new treatment is found to be inferior to the control at an interim analysis, there may still be a reasonable chance of demonstrating non-inferiority in the final analysis. To explore the issues arising from trials where a simultaneously inferior and non-inferior result could occur and to describe appropriate methods for deciding whether such trials should be stopped for futility at an interim analysis. Conditional power is used to assess futility or the inability of the trial to show non-inferiority at the final analysis, by calculating the probability of demonstrating non-inferiority in the final analysis conditional on the observed results and upon assumptions on the future results of the trial. The Bullous Pemphigoid Steroids and Tetracyclines Study (BLISTER) trial is an example of a trial where a simultaneous inferior and non-inferior result could occur. A method for calculating conditional power for non-inferiority using simulations is described and applied at a hypothetical interim analysis of this trial. Stopping guidelines for futility based on conditional power are shown to be better suited to non-inferiority trials than the typical methods used in superiority trials. Simulations are a straightforward and flexible way of calculating conditional power. Calculating conditional power relies on assumptions about future treatment efficacy, and therefore, a number of different conditional power values can be obtained. Careful consideration should be given to which assumptions are most likely to be true. Additionally, when choosing a stopping guideline for futility, consideration needs to be given to avoid overinflating the type II error rate. Conditional power is an appropriate tool for defining stopping guidelines for futility in non-inferiority trials, particularly those with large non-inferiority margins.


Osteopontin expression in activated glial cells following mechanical- or toxin-induced nigral dopaminergic cell loss.
Osteopontin (OPN) is a glycosylated phosphoprotein that regulates both oxidative stress and inflammatory processes. OPN is present in the rat substantia nigra (SN) and both protein and mRNA levels are up-regulated following a pro-inflammatory insult produced by lipopolysaccharide. We now report on the effects of lesioning the SN using 6-hydroxydopamine (6-OHDA) and mechanical vehicle-induced lesioning on OPN expression. Intranigral administration of 6-OHDA induced a marked time-dependent loss of tyrosine hydroxylase (TH) positive nigral cells. Vehicle administration also produced a loss of TH positive cells. This was small compared to 6-OHDA and due to mechanical damage during surgery. 6-OHDA and mechanical-induced cell loss was accompanied by an increase in OPN protein and mRNA expression. Both 6-OHDA and mechanical lesions resulted in equivalent time-dependent increases in OX-42 positive microglial cells. However, the elevation was far less marked following mechanical damage compared to 6-OHDA-induced cell death. 6-OHDA lesioning induced a slow up-regulation of GFAP positive astroglial cells but this was not present following mechanical damage. Importantly, both 6-OHDA and mechanical lesions resulted in an up-regulation in ED1 positive macrophages of equivalent magnitude and time course. There was co-localisation of OPN with ED1 positive cells but not TH, OX-42 or GFAP cells following both toxin and mechanical lesions. Nigral TH positive cell death of toxin or mechanical origin increases OPN expression in parallel with the up-regulation of ED1 positive macrophages. The increase in OPN/ED1 expression is independent of the extent of cell death. OPN appears to be an important regulator of nigral cell survival through its association with inflammatory events and its manipulation may provide a means of achieving neuroprotection in Parkinson's disease.


Joint injury, repair, and remodeling: roles in post-traumatic osteoarthritis.
Joint injuries, especially intraarticular fractures, frequently lead to progressive joint degeneration that causes the clinical syndrome of posttraumatic osteoarthritis. Orthopaedists try to prevent this disease by attempting to restore joint congruity, alignment, and stability; however, many patients have crippling joint pain and dysfunction develop despite optimal current treatment. The pathophysiology of posttraumatic osteoarthritis has not been explained. It is not simply the magnitude and type of injury that determines whether an injured articular surface will repair and remodel or undergo progressive degeneration. For these reasons, clinically significant progress in preventing posttraumatic osteoarthritis depends on advances in understanding of the pathogenesis of this disease that will make it possible to decrease the risk of articular surface degeneration and facilitate articular surface repair and remodeling. We examine the relationships between joint injury, repair and remodeling, and joint degeneration; the factors that increase the risk of posttraumatic joint degeneration; and, the questions that need additional investigation to develop treatments of joint injuries that will decrease the risk or severity of posttraumatic osteoarthritis.


Modification of expression and cis-regulation of Hoxc8 in the evolution of diverged axial morphology.
Differential Hox gene expression between vertebrate species has been implicated in the divergence of axial morphology. To examine this relationship, we have compared expression and transcriptional regulation of Hoxc8 in chicken and mouse. In both species, expression of Hoxc8 in the paraxial mesoderm and neural tube is associated with midthoracic and brachial identities, respectively. During embryogenesis, there is a temporal delay in the activation of Hoxc8 in chicken compared with mouse. As a result, chicken Hoxc8 expression in the paraxial mesoderm is at a posterior axial level, extending over a smaller domain compared with mouse Hoxc8 expression. This finding is consistent with a shorter thoracic region in chicken compared with mouse. In addition, the chicken Hoxc8 early enhancer, differing from its mouse counterpart in only a few specific nucleotides, directs a reporter gene expression to a more posterior domain in transgenic mouse embryos. These findings are consistent with the concept that the diversification of axial morphology has been achieved through changes in cis-regulation of developmental control genes.


The physiological role of ascorbate as photosystem II electron donor: protection against photoinactivation in heat-stressed leaves.
Previously, we showed that ascorbate (Asc), by donating electrons to photosystem II (PSII), supports a sustained electron transport activity in leaves in which the oxygen-evolving complexes were inactivated with a heat pulse (49°C, 40 s). Here, by using wild-type, Asc-overproducing, and -deficient Arabidopsis (Arabidopsis thaliana) mutants (miox4 and vtc2-3, respectively), we investigated the physiological role of Asc as PSII electron donor in heat-stressed leaves (40°C, 15 min), lacking active oxygen-evolving complexes. Chlorophyll-a fluorescence transients show that in leaves excited with trains of saturating single-turnover flashes spaced 200 ms apart, allowing continual electron donation from Asc to PSII, the reaction centers remained functional even after thousands of turnovers. Higher flash frequencies or continuous illumination (300 μmol photons m(-2) s(-1)) gradually inactivated them, a process that appeared to be initiated by a dramatic deceleration of the electron transfer from Tyr(Z) to P680(+), followed by the complete loss of charge separation activity. These processes occurred with half-times of 1.2 and 10 min, 2.8 and 23 min, and 4.1 and 51 min in vtc2-3, the wild type, and miox4, respectively, indicating that the rate of inactivation strongly depended on the Asc content of the leaves. The recovery of PSII activity, following the degradation of PSII proteins (D1, CP43, and PsbO), in moderate light (100 μmol photons m(-2) s(-1), comparable to growth light), was also retarded in the Asc-deficient mutant. These data show that high Asc content of leaves contributes significantly to the ability of plants to withstand heat-stress conditions.


Two-portal video-assisted colopexy in the treatment of intussusception and prolapse of the descending colon in a lamb.
A lamb presented with recurrent prolapse of the descending colon. On clinical examination, intussusception of the descending colon with the prolapse of a segment was verified. The external anal sphincter had a rupture, extending to the lacerated wound in the anus. The lamb underwent colopexy with the two-portal video-assisted incisional technique and was discharged 6 days after the surgical procedure with a satisfactory clinical outcome. There were no recurrences or complications for at least 9 months. Video-assisted colopexy is an alternative treatment for intussusception and recurrent colon prolapse in sheep, even in the presence of an external anal sphincter rupture.


Non-canonical genomic driver mutations of urethane carcinogenesis.
The carcinogen urethane induces pulmonary tumors in mice initiated by an incredibly specific Q61L/R oncogenic mutation in the proto-oncogene Kras. Previous Whole-Exome Sequencing of urethane-induced tumors revealed a bias towards A➙T/G and G➙A substitutions. Subsequent ultra-sensitive Maximum-Depth Sequencing of Kras shortly after urethane exposure suggest a further refinement to CA➙CT/G substitutions. As C182AA➙C182T/GA substitutions in Kras result in Q61L/R mutations, the extreme bias of urethane towards these genomic driver mutations can be ascribed to the specificity of the carcinogen for CA➙CT/G substitutions. However, we previously found that changing rare codons to common in the Kras gene to increase protein expression shifted mutations in urethane-induced tumors away from Kras, or when detected in Kras, to G12D mutations that are usually rarely detected in such tumors. Moreover, the loss of p53 partially reversed this effect, generating tumors with either Q61L/R or G12D oncogenic Kras mutations, or no Kras mutations, presumably due to other genomic driver mutations. Determining the origin of these G12D and other unknown non-canonical genomic driver mutations would provide critical insight into the extreme bias of carcinogens for specific genomic driver mutations. We thus compared the types of Single Nucleotide Variations detected by previously performed Maximum-Depth Sequencing immediately after urethane exposure to the mutation signatures derived from Whole Exome Sequencing of urethane-induced tumors. This identified two types of non-canonical mutations. First, a V637E oncogenic mutation in the proto-oncogene Braf that conforms to the mutation signature of urethane, suggesting that the mutational bias of the carcinogen may account for this non-canonical mutation, similar to that for canonical Q61L/R mutations in Kras. Second, G12D and Q61H mutations in Kras that did not fit this mutation signature, and instead shared similarity with Single Nucleotide Variations detected by Maximum-Depth Sequencing from normal cells, suggesting that perhaps these mutations were pre-existing. We thus posit that when canonical Kras mutations are selected against that the carcinogen may instead promote the expansion of pre-existing genomic driver mutations, although admittedly we cannot rule out other mechanisms. Interrogating the mutation signatures of human lung cancers similarly identified KRAS genomic driver mutations that failed to match the mutation signature of the tumor. Thus, we also speculate that the selection for non-canonical genomic driver mutations during urethane carcinogenesis may reflect the process by which discordance between genomic driver mutations and mutational signatures arises in human cancers.


Metal releases from a municipal solid waste incineration air pollution control residue mixed with compost.
The influence of 10 wt.% mature compost was tested on the heavy metal leachate emissions from a calcium-rich municipal solid waste incineration air pollution control residue (MSWI APC). Apart from elongated columns (500 and 1250 mm), an otherwise norm compliant European percolation test setup was used. More than 99% of the metals Al, As, Cd, Cr, Cu, Fe and Ni were left in the APC residue after leaching to a liquid-to-solid ratio (L/S) of 10. Apparent short-term effects of elevated leachate DOC concentrations on heavy metal releases were not detected. Zn and Pb leachate concentrations were one order of magnitude lower for L/S 5 and 10 from the pure APC residue column, which suggests a possible long-term effect of compost on the release of these elements. Prolonging the contact time between the pore water and the material resulted in elevated leachate concentrations at L/S 0.1 to L/S 1 by a factor of 2. Only Cr and Pb concentrations were at their maxima in the first leachates at L/S 0.1. Equilibrium speciation modelling with the PHREEQC code suggested portlandite (Ca(OH)2) to control Ca solubility and pH.


Unaltered membrane properties of arterial muscle in Dahl strain genetic hypertension.
To characterize membrane properties of arterial muscle from Dahl strain hypertensive rats, we measured caudal artery contractile sensitivity to norepinephrine and serotonin, membrane potential at 16 and 37 degrees C, and intracellular potassium, sodium, and chloride content. Dahl salt-resistant (R) strain fed low- or high-salt diets and Dahl salt-sensitive (S) strain fed a low-salt diet remained normotensive. The Dahl S strain on high-salt diets became hypertensive after 4 wk of high salt feeding. There was no significant difference in the norepinephrine or serotonin effective concentration (EC) EC50, EC10, or maximum response between the hypertensive and any of the three normotensive groups. Membrane potential measured at 37 and 16 degrees C and electron-probe analysis of intracellular potassium, sodium, and chloride concentration showed no significant differences between the four groups of animals. These results that arterial muscle membrane mechanisms are not altered in genetically hypertensive Dahl salt-sensitive rats.


Prospective studies in the natural history of food allergy.
The study of the natural history of food allergy is hindered by the lack of a reliable method of diagnosing food allergy except through oral challenge. This need raises a conflict between the need to document continued sensitivity and the effects of these repeated challenges not only on the natural history but also on the immediate induction of symptoms especially in the highly sensitive patient. Despite this difficulty, a number of investigators have provided data on a total of 177 infants and children who were subjected to repeated challenges. In only a few reports were these repeated challenges carried out in rigorous double-blind protocol. When the challenges were repeated, the results did not differ from the group as a whole. These studies agree that the prognosis in food allergy is excellent, with 60% to 73% of infants losing their food sensitivity in the first year and 26% to 53% of older children eventually remitting. The specific food causing symptoms (egg, milk, soy, etc) affected the prognosis but the intensity of IgE-mediated sensitization as indicated by skin testing or RAST did not. The data regarding the loss of immunologic tests as symptoms subsided differed from study to study. This may be due to the age at which patients were first studied varied widely or because the compliance with dietary restrictions in the various studies varied. Finally, a large majority of children developed respiratory symptoms to inhalant allergens at the same time their food-related symptoms disappeared.


Neuroprotective and antioxidant effects of ghrelin in an experimental glaucoma model.
Damage to retinal ganglion cells due to elevation of intraocular pressure (IOP) is responsible for vision loss in glaucoma. Given that loss of these cells is irreversible, neuroprotection is crucial in the treatment of glaucoma. In this study, we investigated the possible antioxidant and neuroprotective effects of ghrelin on the retina in an experimental glaucoma model. Twenty-one Sprague-Dawley rats were randomly assigned to three groups comprising seven rats each. The rats in the control group were not operated on and did not receive any treatment. In all rats in the other groups, IOP was increased by cauterization of the limbal veins. After creation of the IOP increase, saline 1 mL/kg or ghrelin 40 μg/kg was administered intraperitoneally every day for 14 days in the vehicle control group and ghrelin groups, respectively. On day 14 of the study, the eyes were enucleated. Levels of malondialdehyde (MDA), nitric oxide (NO), and nitric oxide synthase-2 (NOS2) in anterior chamber fluid were measured. The retinas were subjected to immunohistochemistry staining for glial fibrillary acidic protein (GFAP), S-100, and vimentin expression. Mean levels of MDA, NO, and NOS2 in the aqueous humor were higher in the vehicle control group than in the control group (P<0.05). Mean levels of MDA, NO, and NOS2 in the ghrelin group did not show a significant increase compared with levels in the control group (P>0.05). Retinal TUNEL and immunohistochemistry staining in the vehicle control group showed an increase in apoptosis and expression of GFAP, S-100, and vimentin compared with the control group (P<0.05). In the ghrelin group, apoptosis and expression of GFAP, S-100, and vimentin was significantly lower than in the vehicle control group (P<0.05). This study suggests that ghrelin has antioxidant and neuroprotective effects on the retina in an experimental glaucoma model.


Pattern of disability among persons who availed half-way home-care services for psychosocial rehabilitation.
There is dearth of studies related to pattern of disability among persons who availed psychosocial rehabilitation services in India. We studied the pattern of disability among persons who availed half-way home-care services for psychosocial rehabilitation. Out of 130 case files of discharged patients, 50 files were randomly selected for data collection. Indian Disability Evaluation and Assessment Schedule was used to assess the pattern of disability in the sample. The study revealed that only one-third (35%) of the residents had disability in self-care, 41% in communication and understanding and 47% in interpersonal relationship. Overall, majority (76%) of the respondents had moderate level of psychiatric disability at the time of discharge from half-way home. There was no significant relationship between gender and type of psychiatric illness with the level of disability. The overall disability correlated positively with the duration of illness (r s=0.39). Three-fourth of the residents who availed half-way home-care services had moderate level of disability.


Epidemiology of injured patients in rural Uganda: A prospective trauma registry's first 1000 days.
Trauma is a leading cause of morbidity and mortality worldwide. Data characterizing the burden of injury in rural Uganda is limited. Hospital-based trauma registries are a critical tool in illustrating injury patterns and clinical outcomes. This study aims to characterize the traumatic injuries presenting to Soroti Regional Referral Hospital (SRRH) in order to identify opportunities for quality improvement and policy development. From October 2016 to July 2019, we prospectively captured data on injured patients using a locally designed, context-relevant trauma registry instrument. Information regarding patient demographics, injury characteristics, clinical information, and treatment outcomes were recorded. Descriptive, bivariate, and multivariate statistical analyses were conducted. A total of 4109 injured patients were treated during the study period. Median age was 26 years and 63% were male. Students (33%) and peasant farmers (31%) were the most affected occupations. Falls (36%) and road traffic injuries (RTIs, 35%) were the leading causes of injury. Nearly two-thirds of RTIs were motorcycle-related and only 16% involved a pedestrian. Over half (53%) of all patients had a fracture or a sprain. Suffering a burn or a head injury were significant predictors of mortality. The number of trauma patients enrolled in the study declined by five-fold when comparing the final six months and initial six months of the study. Implementation of a context-appropriate trauma registry in a resource-constrained setting is feasible. In rural Uganda, there is a significant need for injury prevention efforts to protect vulnerable populations such as children and women from trauma on roads and in the home. Orthopedic and neurosurgical care are important targets for the strengthening of health systems. The comprehensive data provided by a trauma registry will continue to inform such efforts and provide a way to monitor their progress moving forward.


Actigraphic monitoring of movement and rest-activity rhythms in aging, Alzheimer's disease, and Parkinson's disease.
Actigraphy, the long-term assessment of wrist movements, is used in several research fields, among which are included sleep and circadian rhythms. Actigraphs record movements using accelerometers. The present paper addresses some basic problems and their solutions in the actigraphic assessment of movement, motor symptoms, circadian rest-activity rhythms, and nocturnal agitation in healthy elderly and elderly suffering from a neurodegenerative disease (i.e., Parkinson's disease or Alzheimer's disease) and summarizes the results of previous and ongoing research. First, we have investigated how to filter the accelerometer signal in order to minimize the contribution of accelerations induced by positional changes in the gravitational field--a strong source of artefacts. A bandpass filter from 0.5 to 11 Hz appropriately assesses movement induced accelerations while minimizing gravitational artefact. The application of a bandpass filter from 0.25 to 2 or 3 Hz, as is used in some of the commercially available actigraphs, results in artefacts and moreover biases the slower part of the movement spectrum. It is therefore far from optimal for research on aging, which is associated with a generalized motor slowing. Second, we have proposed an alternative to traditional methods of signal processing in actigraphy, in order to assess both the duration and intensity of movements, and in order to distinguish Parkinsonian tremor. Based on this algorithm, new types of actigraphs have been designed. Third, we have proposed sensitive variables in order to quantify rest-activity rhythm disturbances in healthy elderly subjects and Alzheimer patients, who often present with symptoms of nocturnal restlessness. Since, in these subjects, research protocols applying enforced phase shifts or time-free environments are unfeasible and not justifiable from an ethical point of view, the variables were specifically designed to assess the functionality of the circadian timing system from actigraphic recordings made in the natural environment of subjects. Examples of the application of actigraphy are given, including studies on symptom fluctuations and medication responses in Parkinson patients, and studies on circadian rhythm disturbances and possible remedies in elderly and Alzheimer patients.


Clobetasol propionate shampoo 0.05%: a new option to treat patients with moderate to severe scalp psoriasis.
Psoriasis is a chronic, papulosquamous condition that affects up to 2% of the U.S. population. Approximately 50% of patients with psoriasis have involvement of the scalp. This was a multicentre, randomized, vehicle-controlled, double-masked and parallel-group study. The aim was to evaluate the efficacy and safety of clobetasol propionate shampoo, 0.05% versus its corresponding vehicle in subjects aged 12 years and older with moderate to severe scalp psoriasis over a treatment period of 4 weeks. Recurrence of scalp psoriasis was assessed during a two week follow-up period. A total of 142 subjects were treated. Results after 4 weeks demonstrated that clobetasol propionate shampoo, 0.05% was with a similar safety profile significantly more effective than its vehicle. The novel short contact shampoo formulation of clobetasol propionate is convenient and efficacious and minimizes systemic exposure while being efficient, safe and well-tolerated in the treatment of moderate to severe scalp psoriasis.


The incidence of satellite cysts in keratocystic odontogenic tumors.
Renaming of the Odontogenic Keratocyst as the Keratocystic Odontogenic Tumor by the World Health Organization (WHO) is based on the aggressive nature of this lesion. Satellite cysts founded in the walls of the original cysts may give rise to a new lesion formation. The aim of this retrospecitve study was to identify the existence of specific features according incidence of satellite cysts and the pallisading of the basal layer of the epithelium and to establish their mutual correlation. The histopathologic data of Keratocystic Odontogenic Tumor on the basis of new WHO's classification (2005) were analized. Prominent palisade basal cell layer was found in 415 (94.75%) and partially absent palisade basal cell layer in 23 (5.25%) cases. Satellite cysts were presented in prominent palisade basal cell layer in 85 specimens (20.5%) and in cases with partial absent of the palisade basal layer in 3 spicemens (13%). The higher the frequency of pallisading was the higher the frequency of satellite cysts was (p > 0.05).


[Experimental approach to the gene therapy of motor neuron disease with the use of genes hypoxia-inducible factors].
Motor neuron disease (MND), or amyotrophic lateral sclerosis, is a fatal neurodegenerative disorder characterized by a progressive loss of motor neurons in the spinal cord and the brain. Several angiogenic and neurogenic growth factors, such as the vascular endothelial growth factor (VEGF), angiogenin (ANG), insulin-like growth factor (IGF) and others, have been shown to promote survival of the spinal motor neurons during ischemia. We constructed recombinant vectors using human adenovirus 5 (Ad5) carrying the VEGF, ANG or IGF genes under the control of the cytomegalovirus promoter. As a model for MND, we employed a transgenic mice strain, B6SJL-Tg (SOD1*G93A)d11 Gur/J that develops a progressive degeneration of the spinal motor neurons caused by the expression of a mutated Cu/Zn superoxide dismutase gene SOD1. Delivery of the therapeutic genes to the spinal motor neurons was done using the effect of the retrograde axonal transport after multiple injections of the Ad5-VEGF, Ad5-ANG and Ad5-IGF vectors and their combinations into the limbs and back muscles of the SOD1(G93A) mice. Viral transgene expression in the spinal cord motor neurons was confirmed by immunocytochemistry and RT-RCR. We assessed the neurological status, motor activity and lifespan of experimental and control animal groups. We discovered that SOD1(G93A) mice injected with the Ad5-VEGF + Ad5-ANG combination showed a 2-3 week delay in manifestation of the disease, higher motor activity at the advanced stages of the disease, and at least a 10% increase in the lifespan compared to the control and other experimental groups. These results support the safety and therapeutic efficacy of the tested recombinant treatment. We propose that the developed experimental MND treatment based on viral delivery of VEGF + ANG can be used as a basis for gene therapy drug development and testing in the preclinical and clinical trials of the MND.


Using Visual Representations to Demonstrate Complexity in Mixed Emotional Development Across Childhood.
Previous studies have shown a developmental trend in mixed emotional understanding. As children develop throughout childhood, they begin to recognise simultaneity of positive and negative emotions. However, previous studies have limited ecological validity as they assessed emotion choice using only a single positive and single negative emotion. Therefore, the present study aims to broaden the understanding of mixed emotional development by allowing a wider emotion choice. Mixed emotions were measured using the analogue emotions scale (AES) which allows both intensity of the emotional responses and time to be captured. In the present study, 211 children aged 4-10 were divided into one of three protagonist conditions (self, peer and adult) and read a vignette about the protagonist moving house. Choosing from seven emotions (happy, calm, surprise, sad, worry, fear and anger), they plotted the intensity and duration of each emotion they thought was represented in the vignette. The present study replicated the developmental trend that younger children are more likely than older children to choose a single emotion, and older children are more likely to perceive more simultaneity of emotion than younger children. This trend was demonstrated in the number of emotions chosen, and also the complexity of the AES pattern plotted. Additionally, the present study extended previous research by demonstrating that by broadening the emotion choice, the emotion interaction is more complex than previous studies were able to show.


Using model-based geostatistics for assessing the elimination of trachoma.
Trachoma is the commonest infectious cause of blindness worldwide. Efforts are being made to eliminate trachoma as a public health problem globally. However, as prevalence decreases, it becomes more challenging to precisely predict prevalence. We demonstrate how model-based geostatistics (MBG) can be used as a reliable, efficient, and widely applicable tool to assess the elimination status of trachoma. We analysed trachoma surveillance data from Brazil, Malawi, and Niger. We developed geostatistical Binomial models to predict trachomatous inflammation-follicular (TF) and trachomatous trichiasis (TT) prevalence. We proposed a general framework to incorporate age and gender in the geostatistical models, whilst accounting for residual spatial and non-spatial variation in prevalence through the use of random effects. We also used predictive probabilities generated by the geostatistical models to quantify the likelihood of having achieved the elimination target in each evaluation unit (EU). TF and TT prevalence varied considerably by country, with Brazil showing the lowest prevalence and Niger the highest. Brazil and Malawi are highly likely to have met the elimination criteria for TF in each EU, but, for some EUs, there was high uncertainty in relation to the elimination of TT according to the model alone. In Niger, the predicted prevalence varied significantly across EUs, with the probability of having achieved the elimination target ranging from values close to 0% to 100%, for both TF and TT. We demonstrated the wide applicability of MBG for trachoma programmes, using data from different epidemiological settings. Unlike the standard trachoma prevalence survey approach, MBG provides a more statistically rigorous way of quantifying uncertainty around the achievement of elimination prevalence targets, through the use of spatial correlation. In addition to the analysis of existing survey data, MBG also provides an approach to identify areas in which more sampling effort is needed to improve EU classification. We advocate MBG as the new standard method for analysing trachoma survey outputs.


Therapy and prognosis of hypertension in chronic nephritis.
Statistical analysis of the realtion between blood pressure and renal function in 421 patients with CGN, referred to the Second Internal Medicine at Nihon University Hospital, and in 253 Hypertensive patients with CGN by questionaires sent to 29 Medical Universities were investigated. The relationship between survival rate and blood pressure of 84 patients with CGN in Surugadai Nihon University Hospital was also examined. These data show that antihypertensive therapy for CGN with hypertension has an important effect on prognosis. Propranolol was given to 10 hypertensive patients with CGN and hypotensive effect on renal function was observed. Our experience suggests that propranolol may be useful for treating a high renin component in the hypertension with non renal failure, and renal function does not become worse. But in renal failure, propranolol therapy must be used carefully because of inducement to cardiac failure.


Modeling of protein electrophoresis in silica colloidal crystals having brush layers of polyacrylamide.
Sieving of proteins in silica colloidal crystals of millimeter dimensions is characterized for particle diameters of nominally 350 and 500 nm, where the colloidal crystals are chemically modified with a brush layer of polyacrylamide. A model is developed that relates the reduced electrophoretic mobility to the experimentally measurable porosity. The model fits the data with no adjustable parameters for the case of silica colloidal crystals packed in capillaries, for which independent measurements of the pore radii were made from flow data. The model also fits the data for electrophoresis in a highly ordered colloidal crystal formed in a channel, where the unknown pore radius was used as a fitting parameter. Plate heights as small as 0.4 μm point to the potential for miniaturized separations. Band broadening increases as the pore radius approaches the protein radius, indicating that the main contribution to broadening is the spatial heterogeneity of the pore radius. The results quantitatively support the notion that sieving occurs for proteins in silica colloidal crystals, and facilitate design of new separations that would benefit from miniaturization.


[The mechanism of thermo-induced functional uncoupling of the Ca-pump in the sarcoplasmic reticulum of skeletal muscles].
Short-term heating at 45 degrees C of diluted suspensions of rabbit skeletal muscle sarcoplasmic reticulum (SR) vesicles sharply reduces the Ca/ATP ratio without causing any appreciable changes in the Ca-ATPase activity. The mechanism underlying the uncoupling effect of thermoinactivation was investigated. It was shown that the decrease in the effectiveness of Ca2+ accumulation is correlated with the level of Ca2+ accumulated by SR vesicles. At the same time, thermal inactivation has no effect on passive permeability of SR membranes for Ca2+. Ca2+ (100 microM) added to the inactivation medium partly protects the enzyme against uncoupling. Thermal inactivation of the SR Ca-pump is concomitant with a decreased microviscosity of SR membranes as measured by pyrene excimerization. Addition of 100 microM Ca2+ to the thermoinactivation medium prevents the decrement of microviscosity. It was found that N-3-pyrenylmaleimide, a maleimide derivative of pyrene covalently linked to the SH-groups of Ca-ATPase can also form excimers, which testifies to the formation of Ca-ATPase oligomeric complexes in the native membrane. The probability of oligomerization increases with a thermoinduced decrease in membrane microviscosity, which is correlated with the lowered efficiency of SR Ca-pump operation.


Clone heterogeneity in diploid and aneuploid breast carcinomas as detected by FISH.
We investigated the relationship between DNA ploidy and alterations in chromosomes 1, 8, 12, 16, 17, and 18 in 63 breast carcinoma samples by static cytofluorometry and fluorescence in situ hybridization. Thirty specimens were diploid and 33 were aneuploid. In aneuploid samples, the DNA index value ranged from 1.3 to 3.1, with a main peak near tetraploid values. Diploid clones were present in 21 of 33 aneuploid specimens. Fluorescence in situ hybridization analysis showed a heterogeneous degree of alterations in diploid specimens: one sample was normal, 16 samples had one to three chromosome alterations involving mostly chromosomes 1, 16, and 17, and 13 samples an even higher degree of alterations. The 33 aneuploid specimens showed a very high number of signals (four, five, or more). All the investigated chromosomes were affected in 23 of 33 specimens. Alterations in chromosomes 1 and 17 were detected to a similar percentage in diploid and aneuploid samples, whereas chromosome 16 monosomy was more frequent in diploid samples. Overrepresentation of chromosomes 8, 12, 16, and 18 was significantly higher in aneuploid than in diploid samples. Based on these results, we suggest that diploid and aneuploid breast carcinomas are genetically related. Chromosome 1 and 17 alterations and chromosome 16 monosomy are early changes. Allelic and chromosomal accumulations occur during progression of breast carcinoma by different mechanisms. The high clone heterogeneity found in 17 of 33 aneuploid samples could not be completely explained by endoreduplication and led to the suggestion that chromosomal instability concurs with aneuploidy development. This different evolutionary pathway might be clinically relevant because clone heterogeneity might cause metastasis development and resistance to therapy.


A randomized field trial in Oakland, California.
This study evaluates the impact of a civil remedy program (Beat Health) on drug and disorder problems under experimental field trial conditions. Using calls for service data, the authors find statistically significant differences between the control and experimental groups, when drug problem calls prior to the start of the intervention were compared to drug calls during a 12-month follow-up period. The Beat Health sites, especially the residential sites, improved relative to the control sites. By contrast, the control treatment (patrol response) led to significant increases in drug problems, particularly at the commercial properties included in this study. The authors also examined the spatial influences of the Beat Health and patrol responses in catchment areas surrounding each of the 100 study sites. The results show some improvement in the experimental residential sites. The authors found, however, a possible displacement of drug problems in and around both the commercial experimental and control sites. They show that the displacement effect is most notable in the commercial control sites.


Effect of Topical Anesthesia on Pain from Needle Insertion and Injection and Its Relationship with Anxiety in Patients Awaiting Apical Surgery: A Randomized Double-blind Clinical Trial.
We aimed to simultaneously investigate the effects of topical anesthesia on needle insertion and injection pain in the labial mucosa of the maxillary central incisors of patients awaiting apical surgery and to assess the relationship between patients' anxiety and pain scores. Forty-four patients scheduled for apical surgery of the maxillary anterior incisor or canine were included, and all completed the Modified Dental Anxiety Scale (MDAS) questionnaire. One piece of Xylocaine (AstraZeneca, Sodertalje, Sweden) gauze was applied to the right or left side of the labial vestibule below the central incisor according to a randomization process, and 1 piece of water gauze was applied to the contralateral side of the labial vestibule. Each piece of gauze remained in place for 2 minutes. The subjects were asked to rate their pain according to the numeric rating scale immediately after needle insertion and anesthetic solution injection. Topical anesthetic application significantly reduced both insertion- and injection-related pain. Injection pain was significantly higher than insertion pain throughout the experiment. The difference in pain scores between the placebo and topical anesthetic groups was significantly greater for insertion pain than injection pain. The group with higher MDAS scores showed significantly higher pain scores, except for insertion pain reported by the topical anesthetic group, which did not show a significant difference between MDAS score groups. The topical anesthetic was highly effective for both insertion and injection pain during infiltration anesthesia in the maxillary central incisors. Highly anxious patients reported higher pain scores; however, topical anesthetics reduced the effect of anxiety on increasing pain.


Long-term therapy with low-dose isotretinoin for prevention of basal cell carcinoma: a multicenter clinical trial. Isotretinoin-Basal Cell Carcinoma Study Group.
High-dose isotretinoin has been reported to have a prophylactic effect on nonmelanoma skin cancer, although it is associated with significant toxicity. To test the effectiveness of the long-term administration of low-dose isotretinoin in reducing the occurrence of basal cell carcinoma at a new site in patients with previously treated basal cell carcinomas and to measure the toxicity associated with this regimen, we conducted a clinical trial at eight cancer centers. Nine hundred and eighty-one patients with two or more previously confirmed basal cell carcinomas were randomly assigned to receive either 10 mg of isotretinoin or a placebo daily. Patients were followed for 36 months and monitored at 6-month intervals for skin cancer and toxic effects. After 36 months of treatment, no statistically significant difference in either the cumulative percent of patients with an occurrence of basal cell carcinoma at a new site or the annual rate of basal cell carcinoma formation existed between patients receiving isotretinoin and those receiving the placebo. Elevated serum triglycerides, hyperostotic axial skeletal changes, and mucocutaneous reactions were more frequent in the group receiving isotretinoin than in the control group, and these differences were all statistically significant (P less than .001). This low-dose regimen of isotretinoin not only is ineffective in reducing the occurrence of basal cell carcinoma at new sites in patients with two or more previously treated basal cell carcinomas but also is associated with significant adverse systemic effects. The toxicity associated with the long-term administration of isotretinoin, even at the low dose used in this trial, must be weighted in planning future prevention trials.


Contrast-enhanced magnetic resonance angiography of carotid arterial wall in pigs.
This study was designed to investigate the effects of contrast agents on MR images of balloon-injured carotid arteries containing atherosclerotic-like lesions. We have evaluated an intravascular contrast agent, MS-325 (METASYN INC., Cambridge, MA) and an extravascular contrast agent, Optimark, (Mallinckrodt Medical Inc., St. Louis, MO) on MR angiograms obtained 4 weeks after balloon hyperinflation-induced injury of the left common carotid artery in 12 hypercholesterolemic minipigs. High in-plane resolution (.8 x .4 mm2), thin slice (1 mm) time-of-flight gradient echo sequences were used to acquire the MR angiographic images. Vascular lumen definition was compared before and after a single bolus intravenous injection of a contrast agent. Digital subtraction angiograms were obtained from all pigs after MR imaging. High grade stenosis developed in 1 of the 12 pigs and five pigs had complete occlusion of the injured vessel. The remaining pigs exhibited essentially no visible stenoses as assessed either by MR angiography or digital subtraction angiography. The vessel walls of the stenosed and occluded vessels were visible after the injection of either intravascular or extravascular contrast agent. Histologic analyses showed well developed neovascularization in the neointima or occlusive thrombosis. We conclude that the observed contrast-enhanced vessel wall is caused by an increased vascular supply associated with thrombosis and neointimal thickening that leads to an accumulation of contrast agent in the abnormal vessel walls after the injection of the T1-shortening paramagnetic contrast agent.


Protecting eyewitness evidence: examining the efficacy of a self-administered interview tool.
Given the crucial role of eyewitness evidence, statements should be obtained as soon as possible after an incident. This is not always achieved due to demands on police resources. Two studies trace the development of a new tool, the Self-Administered Interview (SAI), designed to elicit a comprehensive initial statement. In Study 1, SAI participants reported more correct details than participants who provided a free recall account, and performed at the same level as participants given a Cognitive Interview. In Study 2, participants viewed a simulated crime and half recorded their statement using the SAI. After a delay of 1 week, all participants completed a free recall test. SAI participants recalled more correct details in the delayed recall task than control participants.


Comparative analysis of core-fucose-binding lectins from Lens culinaris and Pisum sativum using frontal affinity chromatography.
Lens culinaris lectin (LCA) is a useful probe for the detection in serum of a core-fucosylated alpha-fetoprotein, called AFP-L3 fraction, which is a well-known marker for the diagnosis and prognosis of hepatocellular carcinoma. Here we performed a systematic quantitative interaction analysis of LCA and its close homolog, Pisum sativum lectin (PSA), by frontal affinity chromatography with 143 pyridylaminated (PA) glycans including a series of core-fucosylated glycans. Both lectins showed binding affinity to core-fucosylated, mono- and bi-antennary N-glycans, but not to their tri- and tetra-antennary forms, indicating that the addition of the GlcNAc residue at the N-acetylglucosaminyltransferase IV position abrogates the binding affinity. However, their specificities are distinguishable: while LCA showed the highest affinity to the core-fucosylated, agalactosylated, bi-antennary N-glycan (K(a)=1.1 x 10(5) M(-1)), PSA showed the highest affinity to the core-fucosylated, trimannosyl structure (K(a)=1.2 x 10(5) M(-1)). Glycan-binding specificities of LCA and PSA were also analyzed by glycoconjugate microarray compared to other core-fucose-binding lectins from Aspergillus oryzae (AOL) and Aleuria auratia (AAL). LCA and PSA bound specifically to core fucose, whereas AOL and AAL exhibited broad specificity to fucosylated glycans. These results explain why LCA is appropriate as a specific probe for AFP-L3, which mainly contains a core-fucosylated, biantennary N-glycan, but not its highly branched forms.


Induced pluripotent stem cell-derived cardiac progenitors differentiate to cardiomyocytes and form biosynthetic tissues.
The mammalian heart has little capacity to regenerate, and following injury the myocardium is replaced by non-contractile scar tissue. Consequently, increased wall stress and workload on the remaining myocardium leads to chamber dilation, dysfunction, and heart failure. Cell-based therapy with an autologous, epigenetically reprogrammed, and cardiac-committed progenitor cell source could potentially reverse this process by replacing the damaged myocardium with functional tissue. However, it is unclear whether cardiac progenitor cell-derived cardiomyocytes are capable of attaining levels of structural and functional maturity comparable to that of terminally-fated cardiomyocytes. Here, we first describe the derivation of mouse induced pluripotent stem (iPS) cells, which once differentiated allow for the enrichment of Nkx2-5(+) cardiac progenitors, and the cardiomyocyte-specific expression of the red fluorescent protein. We show that the cardiac progenitors are multipotent and capable of differentiating into endothelial cells, smooth muscle cells and cardiomyocytes. Moreover, cardiac progenitor selection corresponds to cKit(+) cell enrichment, while cardiomyocyte cell-lineage commitment is concomitant with dual expression of either cKit/Flk1 or cKit/Sca-1. We proceed to show that the cardiac progenitor-derived cardiomyocytes are capable of forming electrically and mechanically coupled large-scale 2D cell cultures with mature electrophysiological properties. Finally, we examine the cell progenitors' ability to form electromechanically coherent macroscopic tissues, using a physiologically relevant 3D culture model and demonstrate that following long-term culture the cardiomyocytes align, and form robust electromechanical connections throughout the volume of the biosynthetic tissue construct. We conclude that the iPS cell-derived cardiac progenitors are a robust cell source for tissue engineering applications and a 3D culture platform for pharmacological screening and drug development studies.


A Use of Tritium-Labeled Peat Fulvic Acids and Polyphenolic Derivatives for Designing Pharmacokinetic Experiments on Mice.
Natural products (e.g., polyphenols) have been used as biologically active compounds for centuries. Still, the mechanisms of biological activity of these multicomponent systems are poorly understood due to a lack of appropriate experimental techniques. The method of tritium thermal bombardment allows for non-selective labeling and tracking of all components of complex natural systems. In this study, we applied it to label two well-characterized polyphenolic compounds, peat fulvic acid (FA-Vi18) and oxidized lignin derivative (BP-Cx-1), of predominantly hydrophilic and hydrophobic character, respectively. The identity of the labeled samples was confirmed using size exclusion chromatography. Using ultra-high resolution Fourier transform ion cyclotron resonance mass spectrometry (FT ICR MS), key differences in the molecular composition of BP-Cx-1 and FA-Vi18 were revealed. The labeled samples ([3H]-FA-Vi18 (10 mg/kg) and [3H]-BP-Cx-1 (100 mg/kg)) were administered to female BALB/c mice intravenously (i.v.) and orally. The label distribution was assessed in blood, liver, kidneys, brain, spleen, thymus, ovaries, and heart using liquid scintillation counting. Tritium label was found in all organs studied at different concentrations. For the fulvic acid sample, the largest accumulation was observed in the kidney (Cmax 28.5 mg/kg and 5.6 mg/kg, respectively) for both routes. The organs of preferential accumulation of the lignin derivative were the liver (Cmax accounted for 396.7 and 16.13 mg/kg for i.v. and p.o. routes, respectively) and kidney (Cmax accounted for 343.3 and 17.73 mg/kg for i.v. and p.o. routes, respectively). Our results demonstrate that using the tritium labeling technique enabled successful pharmacokinetic studies on polyphenolic drugs with very different molecular compositions. It proved to be efficient for tissue distribution studies. It was also shown that the dosage of the polyphenolic drug might be lower than 10 mg/kg due to the sensitivity of the 3H detection technique.


Effect of NO inhalation on ECMO use rate and mortality in infants born at or near term with respiratory failure.
The molecular studies showed that Nitric oxide (NO) is an essential factor which regulates pulmonary artery tension. However, the conclusions of existing clinical studies were inconsistent. This meta-analysis is aimed to determine whether the inhalation of NO could improve oxygenation and reduce rate of death and use of extracorporeal membrane oxygenation (ECMO). The strategies used to search PubMed, The Cochrane Central Register of Controlled trials in the Cochrane Library, Embase, Web of science, Clinical Trials Registry, and China Biology Medicine disc, from inception to February, 2018. The primary outcomes were death or use of ECMO, death before hospital discharge, use of ECMO before hospital discharge, change in PaO2 after treatment. We assess the risk of bias in each included study by Cochrane Handbook, and calculated typical estimates of RR, each with its 95% CI, and for continuous outcomes, WMD or a summary estimate for SMD, each with its 95% CI. Nine randomized controlled trials (RCTs) with a total of 856 participants were included in this meta-analysis. This meta-analysis revealed that the experimental group had significantly lower death or use of ECMO (RR 0.66, 95% CI 0.57-0.77, I = 0%, P < .00001) and lower use of ECMO before hospital discharge (RR 0.89, 95% CI 0.50-0.71, I = 0%, P < .00001) compared to control group. And in the infants without diaphragmatic hernia, experimental group had significantly higher change in PaO2 after treatment (MD 50.40, 95% CI 32.14-68.66, P < .00001). The meta-analysis also showing a tendency to improve in the death before hospital discharge (RR 0.89, 95% CI 0.60-1.31, I = 0%, P = .55) and the change in PaO2 after treatment of the infants with diaphragmatic hernia (MD 6.70, 95% CI -2.32 to 15.72, P < .00001, P = .15), but no difference between experimental group and control group. We found that NO inhalation can improve oxygenation and reduce rate of death and use of ECMO in this meta-analysis. Therefore, we recommend the use of NO inhalation for infants born at or near term with respiratory failure.


The Mouse Genome Database (MGD): comprehensive resource for genetics and genomics of the laboratory mouse.
The Mouse Genome Database (MGD, http://www.informatics.jax.org) is the international community resource for integrated genetic, genomic and biological data about the laboratory mouse. Data in MGD are obtained through loads from major data providers and experimental consortia, electronic submissions from laboratories and from the biomedical literature. MGD maintains a comprehensive, unified, non-redundant catalog of mouse genome features generated by distilling gene predictions from NCBI, Ensembl and VEGA. MGD serves as the authoritative source for the nomenclature of mouse genes, mutations, alleles and strains. MGD is the primary source for evidence-supported functional annotations for mouse genes and gene products using the Gene Ontology (GO). MGD provides full annotation of phenotypes and human disease associations for mouse models (genotypes) using terms from the Mammalian Phenotype Ontology and disease names from the Online Mendelian Inheritance in Man (OMIM) resource. MGD is freely accessible online through our website, where users can browse and search interactively, access data in bulk using Batch Query or BioMart, download data files or use our web services Application Programming Interface (API). Improvements to MGD include expanded genome feature classifications, inclusion of new mutant allele sets and phenotype associations and extensions of GO to include new relationships and a new stream of annotations via phylogenetic-based approaches.


Prevalence and Traits of Mobile Colistin Resistance Gene Harbouring Isolates from Different Ecosystems in Africa.
The mobile colistin resistance (mcr) gene threatens the efficacy of colistin (COL), a last-line antibiotic used in treating deadly infections. For more than six decades, COL is used in livestock around the globe, including Africa. The use of critically important antimicrobial agents, like COL, is largely unregulated in Africa, and many other factors militate against effective antimicrobial stewardship in the continent. Currently, ten mcr genes (mcr-1 to mcr-10) have been described. In Africa, mcr-1, mcr-2, mcr-3, mcr-5, mcr-8, and mcr-9 have been detected in isolates from humans, animals, foods of animal origin, and the environment. These genes are harboured by Escherichia coli, Klebsiella, Salmonella, Citrobacter, Enterobacter, Pseudomonas, Aeromonas, Alcaligenes, and Acinetobacter baumannii isolates. Different conjugative and nonconjugative plasmids form the backbone for mcr in these isolates; however, mcr-1 and mcr-3 have also been integrated into the chromosome of some African strains. Insertion sequences (ISs) (especially ISApl1), either located upstream or downstream of mcr, class 1 integrons, and transposons, are drivers of mcr in Africa. Genes coding multi/extensive drug resistance and virulence are colocated with mcr on plasmids in African strains. Transmission of mcr to/among African strains is nonclonal. Contact with mcr-habouring reservoirs, the consumption of contaminated foods of animal/plant origin or fluid, animal-/plant-based food trade and travel serve as exportation, importation, and transmission routes of mcr gene-containing bacteria in Africa. Herein, the current status of plasmid-mediated COL resistance in humans, food-producing animals, foods of animal origin, and environment in Africa is discussed.


Diversity and Plant Growth-Promoting Ability of Endophytic, Halotolerant Bacteria Associated with Tetragonia tetragonioides (Pall.) Kuntze.
The diversity of salt-tolerant cultivable endophytic bacteria associated with the halophyte New Zealand spinach (Tetragonia tetragonioides (Pall.) Kuntze) was studied, and their plant beneficial properties were evaluated. The bacteria isolated from leaves and roots belonged to Agrobacterium, Stenotrophomonas, Bacillus, Brevibacterium, Pseudomonas, Streptomyces, Pseudarthrobacter, Raoultella, Curtobacterium, and Pantoea. Isolates exhibited plant growth-promoting traits, including the production of a phytohormone (indole 3-acetic-acid), cell wall degrading enzymes, and hydrogen cyanide production. Furthermore, antifungal activity against the plant pathogenic fungi Fusarium solani, F. oxysporum, and Verticillium dahliae was detected. Ten out of twenty bacterial isolates were able to synthesize ACC deaminase, which plays a vital role in decreasing ethylene levels in plants. Regardless of the origin of isolated bacteria, root or leaf tissue, they stimulated plant root and shoot growth under 200 mM NaCl conditions. Our study suggests that halophytes such as New Zealand spinach are a promising source for isolating halotolerant plant-beneficial bacteria, which can be considered as potentially efficient biofertilizers in the bioremediation of salt-affected soils.