Two Types of Refutation in Philosophical Argumentation.
In this paper, I highlight the significance of practices of refutation in philosophical inquiry, that is, practices of showing that a claim, person or theory is wrong. I present and contrast two prominent approaches to philosophical refutation: refutation in ancient Greek dialectic (elenchus), in its Socratic variant as described in Plato's dialogues, and as described in Aristotle's logical texts; and the practice of providing counterexamples to putative definitions familiar from twentieth century analytic philosophy, focusing on the so-called Gettier problem. Moreover, I discuss Lakatos' method of proofs and refutations, as it offers insightful observations on the dynamics between arguments, refutations, and counterexamples. Overall, I argue that dialectic, in particular in its Socratic variant, is especially suitable for the philosophical purpose of questioning the obvious, as it invites reflection on one's own doxastic commitments and on the tensions and inconsistencies within one's set of beliefs. By contrast, the counterexample-based approach to philosophical refutation can give rise to philosophical theorizing that is overly focused on hairsplitting disputes, thus becoming alienated from the relevant human experiences. Insofar as philosophical inquiry treads the fine line between questioning the obvious while still seeking to say something significant about human experiences, perhaps a certain amount of what Lakatos describes as 'monster-barring'-a rejection of overly fanciful, artificial putative counterexamples-has its place in philosophical argumentation.


Tenofovir versus entecavir for tertiary prevention of hepatocellular carcinoma in chronic hepatitis B infection after curative therapy: A systematic review and meta-analysis.
Entecavir (ETV) and Tenofovir disoproxil fumarate (TDF) are the first-line drugs for the treatment of chronic hepatitis B virus (HBV). However, the impact of these two antiviral agents on the outcome of HBV-related hepatocellular carcinoma (HCC) after curative therapy remains to be explored. The purpose of the present study was to compare the effect of ETV and TDF on recurrence and mortality after curative treatment for HBV-related HCC. A comprehensive literature search of multiple electronic databases was conducted from 2000 to January 2022 for studies comparing ETV and TDF for HBV-related HCC patients after curative therapy. The adjusted hazard ratios (aHR) were pooled using a random-effects model. A total of nine studies with 5298 patients were included in the final meta-analysis. TDF was associated with a lower risk of HCC recurrence [aHR 0.73, 95% confidence interval (CI) 0.65-0.81] compared to HCC. TDF reduced the risk of late recurrence compared to ETV (aHR 0.58, 95% CI 0.45-0.76) but not early recurrence (aHR 0.88, 95% CI 0.76-1.02). The mortality risk was also lower with TDF compared to ETV (aHR 0.62, 95% CI 0.50-0.77). TDF was associated with a lower risk of recurrence and mortality than ETV after resection or ablation of HBV-related HCC. Further prospective randomized controlled studies are warranted to validate these results.


Preclinical pharmacology of pirmenol.
Pirmenol, a novel pyridinemethanol derivative, is active in a variety of experimental arrhythmic models of diverse etiology and has a favorable therapeutic index compared with other class I agents. Animal pharmacology studies showed that pirmenol is highly efficacious whether the arrhythmias were atrial or ventricular in origin, chemically, mechanically or electrically induced or of the automaticity or reentrant types. The conscious coronary artery-ligated (Harris) dog model best allowed simulation of a variety of clinical situations in which pirmenol could be used either alone or in combination. Pirmenol was highly effective by both the intravenous and oral routes, causing immediate suppression, prevention or termination of cardiac arrhythmias. Preclinical studies in the dog showed an excellent correlation between the dose of pirmenol, plasma levels and antiarrhythmic efficacy. Administration of pirmenol in the dog at intentionally accelerated infusion rates suggested a relatively wide margin of safety for pirmenol compared with other class I agents. Analysis of the pharmacokinetic data led to the modeling of a rapid infusion-slow infusion bolus for sustained intravenous administration, thereby optimizing therapeutic utility. In vitro electrophysiologic studies in dog Purkinje fibers revealed possibly unique differences of pirmenol from other antiarrhythmic agents. It depresses fast and slow response automaticity and its electrophysiologic effects were less variable than other class I drugs over a spectrum of potassium levels. To test the relevance of the in vitro electrophysiologic results, pirmenol's antiarrhythmic efficacy was assessed in several in vivo dog models in which serum potassium was either increased or decreased.(ABSTRACT TRUNCATED AT 250 WORDS)


Base-editing screens illuminate variant effects in human hematopoiesis.
In a recent issue of Cell, Martin-Rufino et al. develop a strategy for performing high-throughput base-editing CRISPR screens coupled with single-cell readouts in the context of human hematopoiesis. Through a series of proof-of-principle experiments, the authors demonstrate the potential of base-editing screens for the study and treatment of hematological disorders.


Dynamics of the protein metabolic response to burn injury.
The protein metabolic response to burn injury was assessed in 17 children aged 7.1 +/- 1.1 years (mean +/- SEM) and a mean burn size of 65 +/- 7% total body surface area (TBSA) during the acute, flow, convalescent, and recovery phases. Stable isotopes of leucine, valine, lysine, and urea were infused in postabsorptive patients in order to measure protein kinetics. The absolute rate of protein breakdown was assessed from the plasma flux of the essential amino acids (EAA), and the rate of urea production (Ra urea) was used as an index of net protein catabolism. Compared to values obtained in recovered patients, the plasma fluxes of all three EAAs were significantly increased (P less than .05), indicating an increased protein breakdown, during the acute, flow, and convalescent phases of injury. Ra urea, however, was only significantly increased during the flow phase (P less than .01), suggesting that protein breakdown was adequately counteracted in the acute and convalescent phases by elevations in protein synthesis but not in the flow phase. The protein kinetic response did not correlate with changes in the metabolic rate since resting energy expenditure (REE) was significantly increased above predicted levels during the acute and flow phases (by 40% and 50%, respectively), and returned to normal in convalescence.(ABSTRACT TRUNCATED AT 250 WORDS)


Optimizing whole blood lymphocyte proliferation in the rat.
Use of whole blood for lymphocyte proliferation experiments have advantages over the usual isolated lymphocyte procedure. We have systematically optimized methodology for whole blood lymphocyte proliferation in male Sprague-Dawley rats. Both phytohemagglutinin (PHA) and concanavalin A (ConA) stimulate lymphocyte blastogenesis in whole blood. The response of rat lymphocytes to ConA at supra-optimum conditions doubled that of PHA in contrast to what is observed in human lymphocytes. Proliferation rates relate non-linearly to concentrations of both mitogens with profiles showing lag, log and decline phases with increasing concentrations. The effect of incubation period followed a similar pattern with an optimum growth rate at 72 h. Whole blood from adrenalectomized rats gave higher proliferation rates than blood from intact normal rats. Advantages of the technique include: use of as little as 8 microliters of blood for each determination; rats need not be killed, the technique is less cumbersome, less time consuming, and cheaper to carry out than the traditional isolated lymphocyte method.


Controlled interlinking of Au and Ag nanoclusters using 4-aminothiophenol as molecular interconnects.
This work describes the formation of interlinked gold and silver nanoclusters at controlled pH using 4-aminothiophenol (ATP) as a molecular interconnect. UV-visible spectra give on intercrystal plasmon resonance band in the region 550-580 nm. The crystalline heteroassembly formation is also evident from the transmission electron microscopic (TEM) images, whereas X-ray photoelectron spectroscopic (XPS) analysis of the aggregates shows the presence of charged -N species, indicating electrostatic interaction of -N with Ag nanoclusters. Furthermore, electrochemical studies of these heteroassembled systems suggest that silver nanoclusters are not fully passivated by the monolayers of ATP and are accessible for redox reactions.


Pylorus-preserving total pancreatectomy for metastatic renal cell carcinoma: a case report.
Resectable isolated multiple metastases to the pancreas from renal cell carcinoma are rare. In this report, we describe a patient with multiple metastases of renal cell carcinoma to the pancreas who was treated with pylorus-preserving total pancreatectomy. The patient was a 58-year-old Asian woman who had undergone right nephrectomy for renal cell carcinoma 20 years earlier. In 2008, she was diagnosed with multiple metastases of renal cell carcinoma to the pancreas by abdominal computed tomography during routine follow-up for renal cell carcinoma. (18)F-2-fluoro-2-deoxyglucose positron emission tomography/computed tomography showed no accumulation in her body other than the pancreas. Because of concerns about her quality of life after total pancreatectomy, she underwent pylorus-preserving total pancreatectomy. After the resection, her control of blood sugar and quality of life were generally satisfactory. She died as a result of gastrointestinal bleeding 35 months after undergoing pancreatectomy. Pancreatic metastasectomy should be considered, even for multiple metastases, when the primary tumor is renal cell carcinoma and the metastatic lesions are isolated.


The effect of inhibitors of insulin processing on generation of insulin intermediate products from human fibroblast as detected by high performance liquid chromatography (HPLC).
To assess the role of various modulators of insulin processing on cell-associated A14-125I-insulin intermediates in human fibroblasts, we have studied the effect of N-ethylmaleimide (NEM), chloroquine, bacitracin, dansylcadavarine, and phenylarsine oxide on generation of these intermediate products with the use of HPLC. NEM completely inhibited generation of intermediate peaks or iodotyrosine. Chloroquine inhibited conversion of A14-125I-insulin to iodotyrosine by about 75 percent and the remaining A14-125I-insulin was not susceptible to acid wash. Bacitracin, dansylcadavarine, and phenylarsine oxide, on the other hand, stimulated formation of intermediate products with concomitant inhibition of iodotyrosine formation. We conclude that there are at least three components of insulin degradation in human fibroblasts. These include the sulfhydryl group inhibitor-sensitive, the intracellular chloroquine-sensitive, and membrane site inhibitor-sensitive components.


Mechanisms of hyperventilation-induced lung injuries in neonatal rats.
The aim of this study was to investigate the mechanism of early inflammatory injury in neonatal ventilator-induced lung injuries (VILI). Newborn rats were randomly assigned to groups and administrated mechanical ventilation with different tidal volumes. Morphological changes in lung tissues were observed, and the levels of interleukin-6 (IL-6), cysteinyl leukotriene mRNA (CysLT1 mRNA), and nuclear factor-κB mRNA (NF-κBp65 mRNA) in lung tissues were analyzed. The ventilation groups exhibited different degrees of inflammatory cell infiltration, which was aggravated as the tidal volume and ventilation time increased. The IL-6 levels of the hyperventilation 5H, conventional ventilation 5H, hyperventilation 3H, control, and normal lung-tissue group were 785.33±39.06, 701.6±33.65, 686.65±46.85, 637.63±40.55, and 635.02±65.78 pg/g, respectively. Hyperventilation increased the levels of IL-6 and NF-κBp65 mRNA as the ventilation time increased, and IL-6 was positively correlated with NF-κBp65 mRNA levels (r=0.72, P<0.01). Longer hyperventilation periods upregulate the level of CysLT1 mRNA. CysLT1 mRNA/GAPDH of the hyperventilation 5H group was 2.14±1.45 (P<0.01). Mechanical ventilation with a large tidal volume can cause VILI, characterized at an early stage by inflammatory responses and particularly by the increased secretion and invasion of inflammatory cytokines and inflammatory cells. The activation of the NF-κB-IL-6 signaling pathway was an important mechanism for the initiation of VILI. Additionally, CysLT1 was involved in the inflammatory VILI damage, and its upregulation occurred later than that of IL-6.


Long-term exposure of zebrafish to bisphenol F: Adverse effects on parental reproduction and offspring neurodevelopment.
Bisphenol F (BPF), an alternative to bisphenol A (BPA) has potential endocrine and reproductive toxicity; however, the effects of environmental concentrations of BPF on the reproductive and developmental toxicity of offspring following parental exposure to BPF remain unclear. In the present study, the effects of life-cycle BPF exposure at environmental concentrations on zebrafish reproduction, offspring growth, and development were investigated. The results showed that the life-cycle of BPF exposure significantly elevated oxidative stress levels, increased gonadal apoptosis, and reduced zebrafish (F0) spawning. Notably, through maternal transfer, BPF exposure significantly affected offspring development. Developmental parameters such as hatching rate, spontaneous movements, heart rate, body length, and locomotor behavior decreased in zebrafish larvae (F1). In addition, the expression levels of genes related to oxidative stress, apoptosis, and neurodevelopment were altered in F1 larvae. Therefore, the present study provides evidence that BPF, even at environmental concentrations, can be potentially adverse in terms of reproductive defects and offspring neurodevelopmental disorders. Therefore, BPF, as a substitute for BPA, is worthy of in-depth evaluation.


Protective role of ascorbic acid in fishes exposed to organochlorine pollution.
Protective role of ascorbic acid on the toxic effect of aldrin pollution in an air-breathing fish Channa punctatus Bloch is reported. Fishes fed with aldrin mixed diet at 0.25% showed 25% mortality in 30 days. Mortality is preceded by certain changes in peripheral blood such as polycythemia and leucocytosis with increased thrombocyte but decreased neutrophil population. However, a massive dose of ascorbic acid (500 mg/100 g diet) was found to considerably neutralize the toxic effect as evidenced by 10-fold decrease in mortality and significantly lowered haematological response. The significance of these studies in the light of physiological role of ascorbic acid as a micronutrient for protection against pollution stress is discussed.


Administration of docosahexaenoic acid before birth and until aging decreases kainate-induced seizures in adult zebrafish.
Docosahexaeonic acid (DHA) is the final compound in the omega-3 polyunsaturated fatty acids (PUFA) synthetic pathway and the most abundant PUFA found in the brain. DHA plays an essential role in the development of the brain, and the intakes in pregnancy and early life affect growth and cognitive performance later in childhood. Recently, it has been proposed that dietary intake of DHA could be a non-pharmacological interventional strategy for the treatment of seizures in humans. However, to date, the experimental approaches to study the antiepileptic effect of DHA have been exclusively restricted to rodent models during short-to-medium periods of treatment. The purpose of the present study was to test the chronic anticonvulsivant effects of DHA supplementation in zebrafish from the pre-spawning stage to aging, taking advantage of our recently described kainate-induced seizure model using this animal. To that end, two groups of adult female zebrafish were fed with standard or 200mg/kg DHA-enriched diets during 1 month previous to the spawning, and offspring subdivided in two categories, and subsequently fed with standard or DHA diets, generating 4 groups of animals that were aged until 20 months. Afterward, KA was intraperitoneally administered and epileptic score determined. All the DHA-enriched groups presented antiepileptic effects compared to the control group, showing that DHA presents an anticonvulsant potential. Among the studied groups, zebrafish fed with DHA from the pre-spawning stage to aging presented the best antiepileptic profile. These results show a neuroprotective benefit in zebrafish fed with DHA-enriched diet before birth and during the whole life.


The influence of cadmium on the biliary excretion of eosine and bromsulphthalein and on microsomal monooxygenase activities in the rat.
Cadmium sulfate x 8/3 H2O (0.3, 0.6, 1.5 and 2.0 mg/kg) administered simultaneously with eosine and bromsulphthalein (120 mumol/kg i.v.) did not significantly change the biliary excretion of the dyes. After a 3 days pretreatment with 2.0 mg/kg CdSO4 i.p. a body weight loss and an increase in the relative liver weight when calculated on body weight were observed together with an enhanced bile flow, but the excretion of the dyes was not markedly influenced. Ethylmorphine N-demethylation and ethoxycoumarin O-deethylation activities as well as microsomal cytochrome P-450 concentration were diminished by about 50%. It can be concluded that in male rats the hepatic microsomal monooxygenase system is more sensitive towards cadmium than the hepatic transport system for organic anions.


RUST FUNGI AND HOST PLANT COEVOLUTION: DO PRIMITIVE HOSTS HARBOR PRIMITIVE PARASITES?
Abstract- The classical view of rust phylogeny is that rusts found on ferns and conifers are primitive, while rusts that parasitize angiosperms are advanced. This belief was based on the theory that primitive hosts harbor primitive parasites; that is, it assumed coevolution (co-speciation) of hosts and parasites. A cladistic analysis of 30 genera and 28 characters representative of the major patterns of rust fungi diversity is presented. The results of this analysis suggest that tropical short-cycle rusts on angiosperms form the cladistically basal group of rusts, while the rusts on conifers and ferns (Melampsoraceae sensu lato) form a nested terminal clade. These results suggest that rusts and their hosts have not undergone a long period of parallel cladogenesis (co-speciation); host transfer has probably been at least as frequent as co-speciation. The cladograms indicate evolutionary trends of spore stages and life history: urediniospores evidently preceded the evolution of aeciospores and pycniospores within Uredinales, and heteroecism is a derived condition which evolved at least several times. This study stresses the importance of making use of independent cladistic analyses of both host and parasite in order to test assumptions of coevolution and host transfer.


Acid polysaccharide content of frog rod outer segments determined by metachromatic toluidine blue staining.
Sulfonation of periodate-oxidized vicinal hydroxyl groups on a polysaccharide backbone allows binding of toluidine blue (aldehyde bisulfite-toluidine blue or ABT staining) with a concurrent metachromatic shift of the dye's absorption peak from 630 nm (monomer) to 580 nm (isolated dimer interaction at vicinal sulfonate groups) or 540 nm (dye polymer interaction). A molar absorptivity of 2.358 +/- 0.134 X 10(4) at 540 nm for polymeric toluidine blue O chloride (TB) aggregates was determined by spectrophotometry of TB bound to hyaluronic acid (HA) and sulfonated glycogen (SG) in water. Microspectrophotometry of ABT stained frog rod outer segments (FROS) showed spectra similar to TB in aqueous HA and SG solutions with absorbances corresponding to 0.063 M dye bound to sugar. Given two dye molecules bound per sugar residue and a rhodopsin concentration of 3.25 mM in FROS, the above indicates 10 stainable sugars per rhodopsin are contained in these cells. Half of these sugars are sensitive to hyaluronidase digestion implying 5 glycosaminoglycan (GAG) repeating units and 5 stainable oligosaccharide sugar residues per rhodopsin in FROS. The GAGs in FROS appear to be primarily HA. Birefringence measurements at 475 nm indicate that this HA and the oligosaccharide of rhodopsin are anisotropically oriented in these cells.


Design and synthesis of α-phenoxy-N-sulfonylphenyl acetamides as Trypanosoma brucei Leucyl-tRNA synthetase inhibitors.
Human African trypanosomiasis (HAT), caused by the parasitic protozoa Trypanosoma brucei, is one of the fatal diseases in tropical areas and current medicines are insufficient. Thus, development of new drugs for HAT is urgently needed. Leucyl-tRNA synthetase (LeuRS), a recently clinically validated antimicrobial target, is an attractive target for development of antitrypanosomal drugs. In this work, we report a series of α-phenoxy-N-sulfonylphenyl acetamides as T. brucei LeuRS inhibitors. The most potent compound 28g showed an IC50 of 0.70 μM which was 250-fold more potent than the starting hit compound 1. The structure-activity relationship was also discussed. These acetamides provided a new scaffold and lead compounds for the further development of clinically useful antitrypanosomal agents.


Binding of Pentagalloyl Glucose to Aortic Wall Proteins: Insights from Peptide Mapping and Simulated Docking Studies.
Pentagalloyl glucose (PGG) is currently being investigated as a non-surgical treatment for abdominal aortic aneurysms (AAAs); however, the molecular mechanisms of action of PGG on the AAA matrix components and the intra-luminal thrombus (ILT) still need to be better understood. To assess these interactions, we utilized peptide fingerprinting and molecular docking simulations to predict the binding of PGG to vascular proteins in normal and aneurysmal aorta, including matrix metalloproteinases (MMPs), cytokines, and fibrin. We performed PGG diffusion studies in pure fibrin gels and human ILT samples. PGG was predicted to bind with high affinity to most vascular proteins, the active sites of MMPs, and several cytokines known to be present in AAAs. Finally, despite potential binding to fibrin, PGG was shown to diffuse readily through thrombus at physiologic pressures. In conclusion, PGG can bind to all the normal and aneurysmal aorta protein components with high affinity, potentially protecting the tissue from degradation and exerting anti-inflammatory activities. Diffusion studies showed that thrombus presence in AAAs is not a barrier to endovascular treatment. Together, these results provide a deeper understanding of the clinical potential of PGG as a non-surgical treatment of AAAs.


REV-ERB nuclear receptors in the suprachiasmatic nucleus control circadian period and restrict diet-induced obesity.
Circadian disruption, as occurs in shift work, is associated with metabolic diseases often attributed to a discordance between internal clocks and environmental timekeepers. REV-ERB nuclear receptors are key components of the molecular clock, but their specific role in the SCN master clock is unknown. We report here that mice lacking circadian REV-ERB nuclear receptors in the SCN maintain free-running locomotor and metabolic rhythms, but these rhythms are notably shortened by 3 hours. When housed under a 24-hour light:dark cycle and fed an obesogenic diet, these mice gained excess weight and accrued more liver fat than controls. These metabolic disturbances were corrected by matching environmental lighting to the shortened endogenous 21-hour clock period, which decreased food consumption. Thus, SCN REV-ERBs are not required for rhythmicity but determine the free-running period length. Moreover, these results support the concept that dissonance between environmental conditions and endogenous time periods causes metabolic disruption.


Intestinal metaplasia of the stomach confined to the fundic gland area. Report of two cases.
Two patients with intestinal metaplasia of the stomach, whose distribution was exclusively confined to the fundic gland area, are presented herein. The first, a 51-year-old male, had been treated for pernicious anemia for 14 years when he was found to have gastric cancer. His serum gastrin level was quite high, whereas his gastric acid output was markedly low. The polypoid cancer in the fornix of the stomach, which had been removed endoscopically, revealed tubular adenocarcinoma with its invasion limited to the mucosa. The resected stomach showed no residual carcinoma but had numerous minute foci of intestinal metaplasia, diffusely distributed but exclusively confined to the fundic gland area, by macroscopic observation using the leucine aminopeptidase-alkaline phosphatase double staining method. The intestinal metaplasias were all of the complete type, and the parietal and chief cells were almost completely lost. The second patient, a 76-year-old male without pernicious anemia, underwent total gastrectomy for two polypoid cancers in the body of the stomach. The resected specimens, in addition to two hyperplastic polyps in the transitional area, showed the same distribution of intestinal metaplasia as seen in the first patient.


Optimisation of ANN topology for predicting the rehydrated apple cubes colour change using RSM and GA.
In this study, an efficient optimisation method by combining response surface methodology (RSM) and genetic algorithm (GA) is introduced to find the optimal topology of artificial neural networks (ANNs) for predicting colour changes in rehydrated apple cubes. A multi-layered feed-forward backpropagation ANN model of algorithms was developed to correlate one output (colour change) to four input variables (drying air temperature, drying air velocity, temperature of distilled water and rehydration time). A predictive model for ANN topology in terms of the best mean squared error (MSE) performance on validation samples was created using RSM. RSM model was integrated with an effective GA to find the optimum topology of ANN. The optimum ANN had minimum MSE when the number of hidden neurons, learning rate, momentum constant, number of epochs and number of training runs were 13, 0.33, 0.89, 3869 and 3, respectively. MSE of optimal ANN topology on validation samples was 0.0072095. It turned out that the optimal ANN topology can be considered as more precise for predicting colour change in the rehydrated apple cubes. Mean absolute error and regression coefficient (R) of the optimal ANN topology were determined as 0.0259 and 0.96475 for training, 0.0399 and 0.95243 for testing and 0.0264 and 0.95151 for validation data sets. The results of the testing model on new samples showed excellent agreement between the actual and predicted data with coefficient of determination R2 = 0.97.


A third rearranged and expressed gene in a clone of cytotoxic T lymphocytes.
In addition to the two previously identified genes rearranged and expressed in a cytotoxic T-lymphocyte clone, we have identified a third gene that is also rearranged and expressed in the same clone. This new gene shows clonal diversity, codes for a polypeptide chain that contains immunoglobulin-like variable and constant domains, carries potential N-glycosylation sites and is a particularly attractive candidate for the gene that encodes the alpha-subunit of the heterodimeric antigen receptor of this T-cell clone.


Non-contractile cells with thin processes resembling interstitial cells of Cajal found in the wall of guinea-pig mesenteric arteries.
Arterial interstitial cells of Cajal (ICC)-like cells (AIL cells) with a multipolar, irregular, elongated shape and with numerous thin (often less than 1 microm), sometimes branching, processes with lengths up to approximately 60 microm were isolated enzymatically from 1st to 7th order branches of guinea-pig mesenteric artery. Some of the processes of AIL cells were growing (average speed approximately 0.15 microm min-1) and their growth was blocked by 10 microM latrunculin B, an inhibitor of actin polymerisation. Staining with BODIPY phalloidin, a fluorescent dye selective for F-actin, showed the presence of F-actin in the processes of AIL cells. Voltage clamp of single AIL cells revealed an inward current that was four times more dense than in myocytes and was abolished by 10 microM nicardipine, and an outward current carried exclusively by potassium ions that was reduced by 1 mM 4-aminopyridine and/or 100 nM iberiotoxin but unaffected by 10 nM dendrotoxin-K. Imaging of intracellular ionised calcium with fluo-4 using a laser scanning confocal microscope showed local or global calcium transients lasting several seconds in approximately 28 % of AIL cells. When membrane current was recorded simultaneously, the calcium transients were found to correspond to long-lasting transient outward currents, which occurred at potentials positive to -40 mV. Unlike myocytes, AIL cells did not contract in response to 1 mM caffeine or 5 microM noradrenaline, although they responded with a [Ca2+]i increase. The segments of intact arteries did not stain for c-kit, a marker of ICCs. Single AIL cells stained positive for vimentin, desmin and smooth muscle myosin. The presence of ICC-like cells is demonstrated for the first time in the media of resistance arteries.


The organization of the bulbar fibre connections to the trigeminal, facial and hypoglossal motor nuclei. II. An autoradiographic tracing study in cat.
In 34 cats 3H-leucine was injected in the pontine and medullary tegmentum. The location of the labelled neurons and the distribution of the labelled fibres and terminals were studied autoradiographically. The findings indicate that the neurons in the bulbar lateral tegmental field (Berman, 1968) represent the main source of the propriobulbar projections to the hypoglossal, facial and motor V nuclei, while those in the medial tegmental field distribute their fibres mainly to the spinal cord. The neurons in the lateral part of the lateral tegmental field give rise to ascending and descending fibres which compose the lateral propriobulbar system, and distribute fibres mainly to ipsilateral bulbar motor nuclei. The neurons in the medial part of the lateral tegmental field compose the medial propriobulbar system, which is organized bilaterally and tends to distribute fibres to the motor nuclei bilaterally. The various neuronal cell groups which project through the medial propriobulbar system to the different motor nuclei bilaterally show relatively less spatial segregation than those which project through the lateral system to these motor nuclei.


Sex determination of human crania using Mastoid triangle and Opisthion-Bimastoid triangle.
In the present study an attempt has been made to establish standards for sex determination from the various direct and indirect measurements of the cranium. A total of 100 cranium (50 of either sex) were measured for nine direct measurements pertaining to Mastoid triangle and Opisthion-Bimastoid triangle. These measurements were used further to calculate four indirect measurements pertaining to the calculation of Opisthion-Bimastoid triangle area and angles. Analysis of data reveals that the male crania exhibit greater values for all the measurements except the angle right Mastoidale-Opisthion-left Mastoidale. The sex difference has been observed to be statistically significant for all the measurements except for the angles of the Opisthion-Bimastoid triangle. Sectioning point was calculated for the diagnosis of sex based on the mean values of these measurements; the accuracy of sex determination varied from measurement to measurement. The highest value for determining sex was obtained for Asterion-Mastoidale length of right side i.e. 80%, followed by Bi-mastoid breadth i.e. 75%. This suggests that these measurements could be used with relatively high degree of accuracy in determining sex of the unknown crania.


Implications of excessive fibrinolysis and alpha(2)-plasmin inhibitor deficiency in patients with severe head injury.
To evaluate the involvement of the fibrinolytic system, especially focused on alpha(2)-plasmin inhibitor, in patients with head injury. This study consisted of 47 patients with isolated blunt head trauma in whom blood sampling could be initiated within 3 hours after injury. Patients were divided into two groups according to Glasgow Outcome Scale score status at 3 months after injury. In Group 1 patients (n = 26), the outcome was characterized as good recovery or moderate disability; in Group 2 patients (n = 21), the outcome was characterized as severe disability, vegetative state, or death. Concentrations of thrombin-antithrombin III complex were greater than 100 microg/L in 39 of 47 patients, and concentrations in Group 2 patients were elevated significantly beyond the concentrations in Group 1 patients. Activities of alpha(2)-plasmin inhibitor in Group 2 were significantly lower than in Group 1 (P < 0.0001). In Group 1 patients, alpha(2)-plasmin inhibitor activity was greater than 60%, while in all but four Group 2 patients, the inhibitor was reduced to less than 60% of normal activity within 3 hours of injury. All patients with alpha(2)-plasmin inhibitor activity less than 60% showed a marked bleeding tendency and/or severe brain edema. Using sandwich enzyme-linked immunosorbent assay, fibrinogen degradation product and fibrin degradation product were measured separately. A significant correlation was apparent between thrombin-antithrombin III complex and fibrinogen degradation product, as well as between the complex and fibrin degradation product. Marked decreases in alpha(2)-plasmin inhibitor were noted only in patients with thrombin-antithrombin III complex concentrations exceeding 500 microg/L. Fibrinolysis and fibrinogenolysis may be involved according to the degree of coagulation activation in the pathophysiology of severe head injury. Decreased activity of alpha(2)-plasmin inhibitor indicated poor prognosis and may be an exacerbating factor in the acute phase of head trauma.


Evaluation of two nutrition education modules for hospital staff members.
Two nutrition education modules for inservice training of hospital food service personnel were evaluated in 16 hospitals. The modules, dealing with cardiovascular disease and diet, contained a leader's guide, a teaching package, audiovisual material, and suggested learning activities. Pretests and post-tests were used to assess knowledge gains resulting from learning sessions. Participants were divided so that one group took the post-tests before the sessions and one group after the sessions. When differences between groups were controlled, the sessions were shown to have a significant positive effect on post-test scores. Three-fourths of the participants rated the sessions as "very good" to "excellent."


Reducing contrast agent residuals in hospital wastewater: the GREENWATER study protocol.
The potential enviromental impact of iodinated (ICAs) and gadolinium-based contrast agents (GBCAs) have recently come under scrutiny, considering the current nonselective wastewater treatment. However, their rapid excretion after intravenous administration could allow their potential recovery by targeting hospital sewage. The GREENWATER study aims to appraise the effective quantities of ICAs and GBCAs retrievable from patients' urine collected after computed tomography (CT) and magnetic resonance imaging (MRI) exams, selecting ICA/GBCA per-patient urinary excretion and patients' acceptance rate as study endpoints. Within a prospective, observational, single-centre, 1-year framework, we will enrol outpatients aged ≥ 18 years, scheduled to perform contrast-enhanced CT or MRI, willing to collect post-examination urine in dedicated canisters by prolonging their hospital stay to 1 h after injection. Collected urine will be processed and partially stored in the institutional biobank. Patient-based analysis will be performed for the first 100 CT and 100 MRI patients, and then, all analyses will be conducted on the pooled urinary sample. Quantification of urinary iodine and gadolinium will be performed with spectroscopy after oxidative digestion. The evaluation of the acceptance rate will assess the "environmental awareness" of patients and will aid to model how procedures to reduce ICA/GBCA enviromental impact could be adapted in different settings. Key points • Enviromental impact of iodinated and gadolinium-based contrast agents represents a growing point of attention.• Current wastewater treatment is unable to retrieve and recycle contrast agents.• Prolonging hospital stay may allow contrast agents retrieval from patients' urine.• The GREENWATER study will assess the effectively retrievable contrast agents' quantities.• The enrolment acceptance rate will allow to evaluate patients' "green sensitivity".


Complete transection of the main bile duct: minimally invasive treatment with an endoscopic-radiologic rendezvous.
Complete transection of the common bile duct (CBD) is a dramatic and often extremely difficult-to-repair event after surgery. Abdominal biliary fluid collection or jaundice is the initial symptom, and ERCP is the determinant for diagnosis. To evaluate the safety and efficacy of a combined endoscopic-radiologic technique for the reconstruction of the CBD. Single-center retrospective study. Tertiary-care center for biliary surgery. This study involved 22 patients with complete transection of the CBD after cholecystectomy. A guidewire is passed in the subhepatic space through the endoscopic approach. A snare loop is advanced from the percutaneous entry site to catch the free end of the wire and then pulled outside the body: a percutaneous biliary-duodenal (PTBD) drainage is put in place. After a new contralateral PTBD, 4 plastic stents are inserted. The stents are removed endoscopically after 8 to 12 months. Success of the rendezvous maneuver, patient recovery, and patient mortality. After a mean follow-up period of 4 years, 16 patients are asymptomatic. Two patients are still under treatment, and 4 patients underwent surgery, as was the surgeon's choice. Single-center, retrospective study with a small population. Interruption of the biliary tree does not represent an indication for an often-difficult surgical treatment, because the CBD is often thin in the presence of biliary peritonitis. However, the condition can be treated with a rendezvous technique. Surgery can be performed in elective conditions or completely avoided when conservative therapy is selected.


Sequence analysis of the hemagglutinin of B/Ann Arbor/1/86, an epidemiologically significant variant of influenza B virus.
Influenza B/Ann Arbor/1/86 is representative of antigenic variants responsible for the most important influenza epidemics during the 1985/1986 season. The nucleotide sequence of the hemagglutinin HA1 region and the deduced amino acid sequence are presented. Compared to the previous winter's vaccine strain B/USSR/100/83, B/AA/1/86 possesses 18 amino acid substitutions and 2 amino acid insertions, 50% of which are distributed within the vicinity of the reported single immunodominant antigenic site.


MDCT of the myocardium: a new contribution to ischemic heart disease.
Despite the progress made in diagnosis and treatment, cardiovascular diseases remain the main cause of death worldwide. Multidetector row computed tomography (MDCT) provides several diagnostic insights, namely assessment of coronary artery anatomy and measurement of left ventricular volume and function. The ability of CT to show myocardial infarcted areas as an enhanced territory was described in the late 1970s in an animal model. This method found a second wind with the arrival of MDCT technology that led to its clinical application. Several authors describe the ability of MDCT to assess myocardial injury both in animals and humans. The MDCT assessment of myocardial late enhancement is based on the same principle as delayed enhancement MRI. The aim of this review is to cover the technical aspects of cardiac MDCT in assessing the myocardium and its potential in diagnosing ischemic heart disease.


Stereoselective synthesis of 1β,25-Dihydroxyvitamin D3 and its 26,27-hexadeuterated derivative.
A mild convergent synthesis of 1β,25-dihydroxyvitamin D3 (3a), a metabolite of vitamin D3, and its C26,27-hexadeuterated derivative (3b) are described. The A-ring and the CD-fragments are constructed from (R)-carvone and Inhoffen-Lythgoe diol, respectively. The triene system is assembled by a Pd(0)-catalyzed process, which involves an enol-triflate (A-ring fragment) and an alkenyl boronate (CD-side chain fragment). Deuterium labeling is introduced at the last step of the synthesis.


Inhibitory effect of Styrax Japonica Siebold et al. Zuccarini glycoprotein (38 kDa) on interleukin-1β and induction proteins in chromium(VI)-treated BNL CL.2 cells.
Chromium(VI) [Cr(VI)] induces chronic inflammation in hepatocytes. Inflammation has been shown to play an important role in tumorigenesis, tumor progression, and metastasis. To examine the effects of the Styrax Japonica Siebold et al. Zuccarini (SJSZ) glycoprotein on inflammation in BNL CL.2 cells, we evaluated the activities of c-Jun NH(2)-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), nuclear factor (NF)-κB (p50 and p65), and inflammation-related factors [cyclooxygenase (COX)-2, inducible nitric oxide syntheses (iNOS) and interleukin (IL)-1β] in Cr-induced BNL CL.2 cells using immunoblot analysis and RT-PCR. We also used two-dimensional gel electrophoresis (2-DE) to compare between treatments. To determine which proteins are induced by Cr(VI), we evaluated total protein lysates using 2-DE. After Cr(VI) treatment, total protein lysates were prepared and resolved by 2-DE. The results obtained from this study demonstrated that the SJSZ glycoprotein (50 μg/ml) inhibits expression of JNK, ERK, NF-κB, and the expression of COX-2, iNOS, and IL-1β. Moreover, the results obtained from 2DE showed that four proteins out of nine proteins were relatively expressed strongly, while the rest of them were relatively appeared weakly on the gel. Taken together, these data indicate that the SJSZ glycoprotein prevents expression of COX-2, iNOS, and IL-1β by blocking NF-κB and MAPKs in Cr(VI)-induced BNL CL.2 cells.


The diagnostic accuracy of selected neurological tests.
The diagnostic value and reliability of selected neurological clinical tests was studied in control subjects with normal neuroimaging (n=42), and subjects with a focal brain lesion (n=38). The items were studied by two examiners blinded to group membership and using standardized protocols, and subsequently by a neurologist who was not blinded to diagnosis. The positive likelihood ratios ranged from 1.06 (pronator drift) to 22.11 (single leg stance with eyes open, while the negative likelihood ratios ranged from 0.47 (tandem gait) to 0.97 (pupil symmetry). Three items (single leg stance - eyes closed - firm surface; single leg stance - eyes open - foam surface; and tandem gait) successfully distinguished between the two groups (odds ratio p<0.05). The inter-rater reliability was generally poor, with only tandem gait showing excellent agreement (kappa [K]=0.92). Tandem gait was the only item to show noteworthy agreement (K=0.93) between the examiners and the neurologist. The tests varied considerably in their ability to detect radiologically demonstrated structural brain lesions, and several items were poorly reproducible, questioning their value as part of a routine neurological examination.


DMS-Seq for In Vivo Genome-wide Mapping of Protein-DNA Interactions and Nucleosome Centers.
Protein-DNA interactions provide the basis for chromatin structure and gene regulation. Comprehensive identification of protein-occupied sites is thus vital to an in-depth understanding of genome function. Dimethyl sulfate (DMS) is a chemical probe that has long been used to detect footprints of DNA-bound proteins in vitro and in vivo. Here, we describe a genomic footprinting method, dimethyl sulfate sequencing (DMS-seq), which exploits the cell-permeable nature of DMS to obviate the need for nuclear isolation. This feature makes DMS-seq simple in practice and removes the potential risk of protein re-localization during nuclear isolation. DMS-seq successfully detects transcription factors bound to cis-regulatory elements and non-canonical chromatin particles in nucleosome-free regions. Furthermore, an unexpected preference of DMS confers on DMS-seq a unique potential to directly detect nucleosome centers without using genetic manipulation. We expect that DMS-seq will serve as a characteristic method for genome-wide interrogation of in vivo protein-DNA interactions.


Anophthalmia, cleft lip/palate, absent vomer bone, nystagmus, and mental-motor retardation: a new syndrome or Fryns "anophthalmia-plus" syndrome?
We report that a 4-year-old boy presented with right unilateral complete cleft lip and palate, right anophthalmos, left congenital nystagmus, absence of the vomer bone, mental-motor retardation, and normal lymphocyte karyotype (46, XY). For reconstruction of the deformities, we performed cleft lip repair by Millard's rotation-advancement technique and planned cleft palate repair. This combination of cleft lip and palate, anophthalmos, congenital nystagmus, absent vomer bone, and mental-motor retardation has not, to our knowledge, previously been described. We suggest that this represents either another case of the rare Fryns "anophthalmia-plus" syndrome or a new syndrome.


A simplified approach to diagnostic rectal biopsy in infants and children.
Ninety-four patients underwent rectal biopsy with a suction capsule during a two year period. This painless procedure was performed in seventy-one patients for diagnosis of aganglionosis (Hirschsprung's disease). In nine patients, no ganglion cells were encountered, and subsequent surgery confirmed the diagnosis of Hirschsprung's disease in each of these patients. In twenty-three patients, biopsies were taken to search for neuronal storage disease. Six specimens contained abnormal material in the cytoplasm of the ganglion cells.


CSNK1D is associated with stemness and invasiveness in glioblastoma.
Glioblastoma (GBM) is the most common primary malignant brain tumor. It has a poor 5-year survival rate, a high recurrence rate, and few therapeutic options. Exploring the molecular processes underlying the formation and progression of GBM, as well as identifying novel therapeutic targets, is critical for improving GBM therapy and prognosis. We extracted primary GSCs (glioblastoma stem cells) from patient-derived samples. Different levels of CSNK1D were evaluated through immunohistochemistry, western blot and real-time PCR assays. A Transwell assay was used to detect invasive ability of cell lines. Tumorsphere formation assay was performed to detect cancer stemness properties. Orthotopic xenograft models were used to evaluate the effect of CSNK1D on GSC tumorigenesis. We found the expression levels of CSNK1D was elevated in GBM tissues compared with normal brain. CSNK1D expression had an increased tendency among WHO grades (G2-G4), and was higher in IDH wildtype group than in mutation group. The prognosis of the CSNK1D high expression group was significantly worse than that of the low expression group. Cox multivariate analysis showed that CSNK1D was also an independent prognostic factor in GBM patients. In primary GBM cells, we observed increased levels of CSNK1D in GSCs compared to non-stem tumor cells (NSTCs). In addition, the change of stemness markers expression and proliferation of GSCs were in coordinate with CSNK1D overexpression or knockdown. Furthermore, CSNK1D could affect the epithelial-mesenchymal transition (EMT) markers and MMPs expression in GSCs. Finally, disruption of CSNK1D expression impairs GSC survival and GBM tumor propagation in orthotopic xenograft models. Our results suggest that CSNK1D correlated with GBM prognosis and might influence the stemness and invasiveness of GSCs, which represented a potential therapeutic target in GBM patients.


A model for learning to segment temporal sequences, utilizing a mixture of RNN experts together with adaptive variance.
This paper proposes a novel learning method for a mixture of recurrent neural network (RNN) experts model, which can acquire the ability to generate desired sequences by dynamically switching between experts. Our method is based on maximum likelihood estimation, using a gradient descent algorithm. This approach is similar to that used in conventional methods; however, we modify the likelihood function by adding a mechanism to alter the variance for each expert. The proposed method is demonstrated to successfully learn Markov chain switching among a set of 9 Lissajous curves, for which the conventional method fails. The learning performance, analyzed in terms of the generalization capability, of the proposed method is also shown to be superior to that of the conventional method. With the addition of a gating network, the proposed method is successfully applied to the learning of sensory-motor flows for a small humanoid robot as a realistic problem of time series prediction and generation.


Isolated regional therapy for advanced extremity soft tissue sarcomas.
Patients presenting with unresectable, large, primary or recurrent extremity soft tissue sarcoma or locally advanced extremity tumors may benefit from treatment options in the form of isolated regional perfusion therapy. Hyperthermic isolated limb perfusion (HILP) and isolated limb infusion (ILI) have proved to be efficacious with acceptable systemic and regional toxicity profiles. Both procedures are attractive as options for patients who might otherwise be facing amputation as limb salvage procedures. HILP and ILI can be offered as either definitive treatment or as neoadjuvant therapy followed by surgery and/or radiation treatment. Response rates are encouraging as are limb preservation rates after regional therapy. Ongoing multicenter collaborations and clinical trials are required to gain knowledge on HILP and ILI for unresectable extremity sarcoma and expand the indications for use in the management of advanced extremity soft tissue sarcoma.


The Rho/ROCK pathway for lysophosphatidic acid-induced proteolytic enzyme expression and ovarian cancer cell invasion.
Lysophosphatidic acid (LPA) is a biolipid that has diverse biological activities implicated in ovarian cancer initiation and progression. Previous studies have shown the critical role of the Rho/Rho-associated kinase (ROCK) pathway in LPA-induced ovarian cancer progression. However, detailed underlying mechanism by which the Rho/ROCK pathway induces ovarian cancer cell invasion is still incompletely understood. In the present study, we observed that the Rho/ROCK pathway is implicated in the production of proteolytic enzymes, leading to LPA-induced ovarian cancer cell invasion. LPA induced matrix metalloproteinase (MMP)-9 expression in CAOV-3 and PA-1 cells and urokinase-type plasminogen activator (uPA) expression in SKOV-3 cells. LPA-induced proteolytic enzyme expression was required for the invasion of ovarian cancer cells expressing corresponding enzymes. Pretreatment of cells with a pharmacological inhibitor of Rho/ROCK (Y-27632) or overexpression of a dominant-negative mutant of Rho (Rho N19) profoundly inhibited LPA-induced proteolytic enzyme expression as well as the invasive potential of ovarian cancer cells. In addition, transfection with dominant-negative Ras (Ras N17) significantly inhibited LPA-induced Rho activation as well as MMP-9 and uPA expression. Consistently, Y-27632 reduced LPA-induced nuclear factor (NF)-κB activation that is critical for proteolytic enzyme expression and cellular invasion. Collectively, we demonstrate a mechanism by which LPA promotes ovarian cancer progression through coordinate activation of a Ras/Rho/ROCK/NF-κB signaling pathway and the proteolytic enzyme secretion, providing novel biomarkers and promising therapeutic targets for ovarian cancer cell progression.


Pyruvate dehydrogenase activity in the rat cerebral cortex following cerebral ischemia.
The effect of cerebral ischemia on the activity of pyruvate dehydrogenase (PDH) enzyme complex (PDHC) was investigated in homogenates of frozen rat cerebral cortex following 15 min of bilateral common carotid occlusion ischemia and following 15 min, 60 min, and 6 h of recirculation after 15 min of ischemia. In frozen cortical tissue from the same animals, the levels of labile phosphate compounds, glucose, glycogen, lactate, and pyruvate was determined. In cortex from control animals, the rate of [1(-14)C]pyruvate decarboxylation was 9.6 +/- 0.5 nmol CO2/(min-mg protein) or 40% of the total PDHC activity. This fraction increased to 89% at the end of 15 min of ischemia. At 15 min of recirculation following 15 min of ischemia, the PDHC activity decreased to 50% of control levels and was depressed for up to 6 h post ischemia. This decrease in activity was not due to a decrease in total PDHC activity. Apart from a reduction in ATP levels, the acute changes in the levels of energy metabolites were essentially normalized at 6 h of recovery. Dichloroacetate (DCA), an inhibitor of PDH kinase, given to rats at 250 mg/kg i.p. four times over 2 h, significantly decreased blood glucose levels from 7.4 +/- 0.6 to 5.1 +/- 0.3 mmol/L and fully activated PDHC. In animals in which the plasma glucose level was maintained at control levels of 8.3 +/- 0.5 mumol/g by intravenous infusion of glucose, the active portion of PDHC increased to 95 +/- 4%. In contrast, the depressed PDHC activity at 15 min following ischemia was not affected by the DCA treatment.(ABSTRACT TRUNCATED AT 250 WORDS)


Application of noninvasive positive pressure ventilation to respiratory complications of severe tetanus: a case report.
Respiratory failure because of muscle spasms is a major sequela of tetanus. Spasms can be controlled with sedation and adjunctive treatment, albeit at the expense of respiratory drive and weakness. Invasive ventilation under deep sedation with intensive care unit admission is the best practice. Where invasive ventilation is unavailable and when patients decline intubation, the management of muscle spasms and respiratory failure is extremely limited, with low survival rates. Herein, we present a case of severe tetanus in a 90-year-old female farmer who declined intubation and whose respiratory failure was managed with noninvasive positive pressure ventilation (NPPV) with a moderate dose of continuous benzodiazepine infusion to treat spasms. Compared with invasive ventilation, NPPV reduces the duration of ventilatory support and the risk of nosocomial pneumonia. Because spontaneous respiratory drive of the patient was maintained to trigger ventilation, pressure control ventilation was initiated. In previous references to tetanus management, the application of NPPV has not been mentioned. NPPV is a treatment option for patients with tetanus who are not intubated owing to limited medical resources or do-not-intubate orders for older patients.


Result of two randomized trials comparing nolatrexed (Thymitaq) versus methotrexate in patients with recurrent head and neck cancer.
We report on two randomized trials performed in the USA and Europe, which compared methotrexate and nolatrexed as treatment for patients with recurrent head and neck cancer. Eligibility criteria included: histologically confirmed squamous-cell carcinoma, measurable disease, adequate hematological, renal and hepatic functions, failure of a first-line chemotherapy, and informed consent. Methotrexate 40 mg/m2 was weekly given by short infusion, and nolatrexed 725 mg/m2 per day was administered as a five-day continuous infusion, every three weeks. A total of 139 patients (63 in the USA. 76 in Europe) were randomized based on a ratio of 2/1: 93 and 46 received nolatrexed and methotrexate, respectively. Patient characteristics included 115 males and 24 females; median age 60 years. In the nolatrexed arm, the following grade 3-4 toxicities occurred: neutropenia (29.9%) with 3.1% of febrile neutropenia, mucositis (33.3%), and vomiting (10.3%). In the MTX arm, the grade 3-4 toxicities were neutropenia (7.1%) and mucositis (6.9%). There was no difference in activity between the nolatrexed and the methotrexate treatment: 3.3% and 10.8% of objective responses, 1.9 versus 1.5 months of disease-free progression and 3.5 versus 3.7 months of overall survival, respectively. Nolatrexed has demonstrated a similar activity to methotrexate.


Fluctuations in astrocytic basic fibroblast growth factor in the cingulate cortex of cycling, ovariectomized and postpartum animals.
In the current studies we investigated the timing of onset and the conditions needed for the maintenance of the upregulation of basic fibroblast growth factor (bFGF) and glial fibrillary acidic protein (GFAP) in the cingulate cortex area 2 (Cg2) that occurs in postpartum animals. We have previously shown that this upregulation is present from day 4 to day 24, and is not seen late in pregnancy (days 21-22). In the current studies, we demonstrate that bFGF and GFAP are both upregulated in Cg2 as early as 3 h postpartum, and are maintained until at least day 24 postpartum in animals deprived of pup stimulation for 8 days prior to perfusion. bFGF and GFAP immunoreactivity returns to prepartum levels by 5-6 weeks post-weaning, and the typical postpartum increase is not further enhanced in multiparous rats. We also show that, although there are significant changes in levels of bFGF immunoreactivity across the phases of the estrous cycle, peak cycling levels remain much lower than those observed in lactating rats. Possible stimuli involved in the induction of bFGF and GFAP in Cg2, and the potential relevance of these changes to the maternal state are discussed.


Management Issues in Conjunctival Tumours: Conjunctival Melanoma and Primary Acquired Melanosis.
Conjunctival melanoma is a rare malignant condition of the ocular surface. It is potentially lethal, with regional lymph node spread often preceding distant solid-organ metastasis. Due to its rarity and the long latency between treatment and local recurrence or the development of metastases, it is difficult to study. The literature is composed entirely of case series of varying quality, and there is significant variability in the management of this condition. This commentary attempts to distil the evidence base to provide practical management tips for the clinician.


Immunological responsiveness and adjunct immunotherapy in lung cancer.
Immunocompetency was assessed before and after the operation in 40 patients with lung cancer by skin reaction against tuberculin (PPD) and dinitrochlorobenzene (DNCB), lymphocyte response to PHA, proportion of T-cells, macrophage migration inhibition test (MIT) and the presence of blocking factor. MIT was positive in 27 per cent and blocking factor was positive in 42 per cent. Immune response paralleled the clinical stage of the lesion. In curative resection cases, the immune response rose postoperatively, but declined in non-resectable or recurrent cases. The influence of postoperative radiation therapy, cancer chemotherapy and host mediated agents on the patients was observed. The feasibility of adjuvant specific immunotherapy is discussed.


Biological responses of juvenile European sea bass (Dicentrarchus labrax) exposed to contaminated sediments.
Multiple anthropogenic activities present along coastal environments may affect the health status of aquatic ecosystems. In this study, specimens of European sea bass (Dicentrarchus labrax) were exposed for 30 days to highly contaminated sediment collected from the industrial area between Augusta and Priolo (Syracuse, Italy), defined as the most mercury polluted site in the Mediterranean. The aim was to evaluate the responses of juvenile D. labrax to highly contaminated sediments, particularly enriched in Hg, in order to enhance the scarce knowledge on the potential compensatory mechanisms developed by organisms under severe stress conditions. Apoptotic and proliferative activities [cell turnover: Proliferating Cell Nuclear Antigen (PCNA) and FAS Ligand (FasL)], onset of hypoxic condition [hypoxia: Hypoxia Inducibile Factor-1α (HIF-1α)], and changes in the neuroendocrine control mechanisms [neurotransmission: Tyrosine Hydroxylase (TH), Choline Acetyltransferase (ChAT), Acetylcholinesterase (AChE), 5-Hydroxytryptamine (5-HT) and 5-Hydroxytryptamine receptor 3 (5-HT3)] were investigated in sea bass gill tissues. In the specimens exposed to the polluted sediment, the occurrence of altered cell turnover may result in impaired gas exchange that leads to a condition of "functional hypoxia". Changes in neurotransmission pathways were also observed, suggesting a remodeling process as an adaptive response to increase the O2-carrying capacity and restore the normal physiological conditions of the gills. Overall, these findings demonstrated that although chronic exposure to heavy metal polluted sediments alters the functioning of both the nervous and endocrine systems, as well as plasticity of the gill epithelium, fish are able to trigger a series of physiological adjustments or adaptations interfering with specific neuroendocrine control mechanisms that enable their long-term survival.


Silk nanofibrils-MOF composite membranes for pollutant removal from water.
Membrane separation technology is considered an effective strategy to remove pollutants in sewage. However, it remains a significant challenge to fabricate inexpensive membranes with high purification efficiency. Therefore, the present study proposes the integration of silk nanofibrils (SNFs) and polydopamine⊂metal-organic framework (PDA⊂MOF) nanoparticles to prepare self-supporting membranes, which can effectively intercept nanoparticle pollutants through the size exclusion effect and can strongly adsorb organic dyes and metal ions by SNF. In addition, PDA⊂MOF enables these membranes to adsorb small molecules and heavy metal ions during the filtration process, thereby effectively removing various pollutants from sewage. The integration of size-exclusion and adsorption capabilities enables the SNF/PDA⊂MOF membrane to remove nanoparticles, small-molecule dyes, heavy metal ions, and radioactive elements. This work provides a rational approach for the design and development of the next generation of water treatment membranes and is expected to be used in environmental, food-related, and biomedical fields.


[Gly14]-humanin restores cathepsin D function via FPRL1 and promotes autophagic degradation of Ox-LDL in HUVECs.
Abnormal aggregation of oxidized low-density lipoprotein (Ox-LDL) in vascular endothelial cells (VECs) is one of the major pathological changes in atherosclerotic lesions. Our research aimed to assess the mechanism of humanin (HN) in promoting autophagic degradation of Ox-LDL in HUVECs. Flow cytometry and lipid quantitation results showed that Ox-LDL caused lipid and cholesterol accumulation in HUVECs. Western blot results showed that Ox-LDL increased the expression of autophagy-related proteins P62 and LC3-II in a concentration-dependent manner. The cathepsin D activity assay showed that Ox-LDL inhibited the function of cathepsin D. HNG pretreatment reduced lipid and cholesterol aggregation in HUVECs induced by Ox-LDL, increased LC3-II protein level, decreased P62 protein content, and reversed Ox-LDL-induced cathepsin D functional impairment. Inhibition of the FPRL1 pathway by FPRL1 siRNA or the FPRL1-specific inhibitor Boc-MLF blocked all HNG's protective effects. These results indicate that HNG could restore cathepsin D activity and protein level in HUVECs to repair lysosomal functional damage induced by Ox-LDL, further repairing Ox-LDL-induced autophagic damage in HUVECs. HNG restores the activity of Ox-LDL-induced damaged lysosomal enzyme cathepsin D through its membrane protein receptor FPRL1 to promote autophagic degradation of Ox-LDL in HUVECs.


Fluoride-Induced Alteration in the Diversity and Composition of Bacterial Microbiota in Mice Colon.
Fluoride, as an environmental toxin, causes damage to intestinal mucosa. It may promote pathogen infection by increasing the intestinal mucosa permeability. In this study, the colonic fecal samples from the control group (C group, 0 mg/L NaF for 60 days) and the fluoride group (F group, 100 mg/L NaF for 60 days) were subjected to high-throughput 16S rRNA sequencing to verify the effects of fluoride on the colonic flora of animals. Results revealed a total of 253 operative taxonomical units (OTUs) in two groups, and 22 unique OTUs occurred in the F group. Fluoride increased the microbiota diversity and species richness of the colon. Concretely, the abundance of the Tenericutes was increased at the level of the phyla in the F group. In addition, in the F group, significant differences at the genus level were observed in Faecalibaculum, Alloprevotella, [Eubacterium]_xylanophilum_group, Prevotellaceae_UCG-001, and Ruminiclostridium_9, compared to the C group. Among them, except for the reduction in Faecalibaculum, the other four bacteria were increased in the F group. In summary, the intestinal microbial composition of mice was reconstituted by the presence of fluoride, and the significantly changing bacteria may partly account for the pathogenesis of fluoride-induced intestinal dysfunction.


TRAPnSeq allows high-throughput profiling of antigen-specific antibody-secreting cells.
Following activation by cognate antigen, B cells undergo fine-tuning of their antigen receptors and may ultimately differentiate into antibody-secreting cells (ASCs). While antigen-specific B cells that express surface receptors (B cell receptors [BCRs]) can be readily cloned and sequenced following flow sorting, antigen-specific ASCs that lack surface BCRs cannot be easily profiled. Here, we report an approach, TRAPnSeq (antigen specificity mapping through immunoglobulin [Ig] secretion TRAP and Sequencing), that allows capture of secreted antibodies on the surface of ASCs, which in turn enables high-throughput screening of single ASCs against large antigen panels. This approach incorporates flow cytometry, standard microfluidic platforms, and DNA-barcoding technologies to characterize antigen-specific ASCs through single-cell V(D)J, RNA, and antigen barcode sequencing. We show the utility of TRAPnSeq by profiling antigen-specific IgG and IgE ASCs from both mice and humans and highlight its capacity to accelerate therapeutic antibody discovery from ASCs.


Paracrine signalling in colorectal liver metastases involving tumor cell-derived PDGF-C and hepatic stellate cell-derived PAK-2.
In a nude mouse model of colorectal liver metastases, we have identified a paracrine tumor cell/host cell signalling pathway that is apparently required for successful tumor growth. Whereas recombinant platelet derived growth factor-C (PDGF-C) and supernatants from PDGF-C secreting wild type LS174T colon carcinoma cells could rescue tumor promoting hepatic stellate cells (HSC) from growth inhibition by serum starvation, supernatants from LS174T colon carcinoma cells with reduced secretion of PDGF-C had much less effect on serum starved HSC. Autocrine growth inhibition of LS174T cells by PDGF-C knock-down was only marginal. In vivo, a prominent inhibition of liver metastasis was observed if PDGF-C was knocked-down in LS174T cells. By whole genome array analysis of host cells of the invasion front and subsequent immunohistochemical staining we identified p21 activated kinase-2 (PAK-2) as being strongly and specifically expressed by HSC. The above described effect of PDGF-C on HSC was found to be dependent on PAK-2 because in contrast to wild type HSC, silencing of PAK-2 in HSC only allowed for a partial PDGF-C-mediated rescue from serum starvation leading to only a slight increase of proliferation. These data indicate that PDGF-C promotes tumor growth via a growth promoting effect on HSC that is at least in part dependent on the presence of functional PAK-2.


Infant-feeding practices in Western Australia and Tasmania: a joint survey, 1984-1985.
A joint survey of infant-feeding practices that was carried out in Western Australia and Tasmania in 1984-1985 showed a continuing trend back to breast-feeding in both States. In Western Australia and Tasmania, 86% and 81% of mothers, respectively, were breast-feeding their babies on hospital discharge. Forty-five per cent of all mothers were still breast-feeding at six months. The social rank of the family had a significant effect on both the prevalence of breast-feeding and on the length of lactation: more mothers in the higher social groups breast-fed their infants, and for longer periods than did mothers of lower social groups. Few infants were introduced to solid foods before three months of age; however, solid and non-milk foods were introduced earlier to infants who were fed artificially than those who were breast-fed.


Determination of toxaphene specific congeners in fish liver oil and feedingstuff using gas chromatography coupled to high resolution mass spectrometry.
A new method for the determination of nine toxaphene specific congeners in fish liver oil and feedingstuff has been developed. The samples were extracted using pressurized liquid extraction followed by a purification on silica and florisil columns. Identification and quantification were conducted using GC-(EI)-HRMS, and comparison with MS/MS detection was performed, using electron ionization and negative chemical ionization. Limits of detection were ranged from 0.01 to 0.22 microg kg(-1) (12% moisture) as required for feed samples. The calibration curves showed a good linearity for all congeners (R(2)>0.99). Repeatability was below 9% for all the congeners and recoveries were in-between 73 and 86%. This analytical method was applied to the quantification of thirteen real samples collected within national monitoring plans for further risk assessment.


Updates on Therapeutic Strategies Against Candida (and Aspergillus) Biofilm Related Infections.
Fungal biofilm related infections are commonly associated with medical devices with biofilms contributing to the virulence of the involved fungal species. If infection does occur, removal of medical device is often warranted. However, this is not always possible. Moreover, biofilm associated infections are often resistant to antifungals and host immunity. Therefore, a need for new agents and strategies to combat these devastating infections is needed. Although no randomized clinical trials have been conducted or are likely to be conducted in the future, the Infectious Disease Society of America (IDSA) and the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) utilized available published data and clinical experience of the infectious disease community to propose strategies to treat biofilm associated devise infections. In this chapter we describe the emerging therapies for biofilm related infections.


Diesel exhaust exposure and lung cancer: a case--control study.
The presence of polyaromatic hydrocarbons in the particulate phase of diesel engine exhaust has raised questions concerning potential carcinogenicity of diesel exhaust exposure. A case-control study was conducted of 502 male lung cancer cases and 502 controls without tobacco-related diseases to investigate the association of occupational diesel exhaust exposure and lung cancer. Diesel exhaust exposure was appraised by job title. The results show no association between diesel exhaust exposure and risk of lung cancer. They do, however, show the strong association between smoking and lung cancer and as such highlight the importance of smoking information in studies of occupational effect on lung cancer risk.


Diethylmaleate, a pro-oxidant, attenuates experimental ischaemia-reperfusion-induced lung injury.
Systemic ischaemia-reperfusion (IR) injury is in part an oxidant injury mediated by neutrophils. Diethylmaleate (DEM), an intracellular pro-oxidant agent, has been shown to alleviate neutrophil-mediated tissue injury. The aim of this study was to evaluate whether DEM could have a protective effect on neutrophil-mediated lung injury in an animal model of lower-torso IR. Sprague-Dawley rats (seven per group) were randomized into three groups. The control group underwent midline laparotomy only; the IR group underwent laparotomy and clamping of the infrarenal abdominal aorta for 30 min followed by 2 h of reperfusion; and the third group was pretreated with DEM 6 mmol/kg intraperitoneally 1 h before the IR insult. IR resulted in a significant increase in both microvascular leakage and pulmonary neutrophil infiltration as measured by bronchoalveolar lavage protein concentration and pulmonary myeloperoxidase activity respectively. Pretreatment with DEM significantly attenuated both microvascular leakage and neutrophil infiltration. Preconditioning with DEM protected against IR-induced lung injury. This protective effect raises the possibility of using pro-oxidants to prevent inflammatory injury.


Clinical efficacy evaluation of a traditional Miao technique of crossbow needle therapy in the treatment of knee osteoarthritis: a multi-center randomized controlled trial.
Knee osteoarthritis (KOA) seriously reduces quality of life and is a major threat to the health of the middle-aged and elderly. This study aimed to assess the efficacy of Miao crossbow needle therapy vs. acupuncture for KOA therapy. This multicenter, randomized controlled trial was performed at three hospitals between April 2016 and December 2016. The patients were randomized to receive crossbow needle (CN) or acupuncture (AT). All treatments were completed within 46 days. Evaluation of treatment was conducted on the 46th, 62nd, and 77th days. The primary endpoint was change of Western Ontario and McMaster Osteoarthritis Index (WOMAC) score on the 46th day. The secondary endpoints included WOMAC score, the Lysholm knee score, the Japanese Orthopedic Association (JOA) knee score, visual analog scale (VAS), and the MOS 36-item short-form health survey (SF-36), on the 46th, 62nd, and 77th day. Finally, data of 301 participants were analyzed for the efficacy of treatment. Compared with AT, there was a larger change of WOMAC score in the CN group after treatment [- 25.0 (95% CI - 27.0, - 23.0) vs. - 18.8 (95% CI - 20.8, - 16.9), P < 0.001]. In the CN group, the WOMAC score was lower at all three time points (P = 0.008, P = 0.003, P < 0.001 respectively), while the Lysholm knee score (P = 0.03) and JOA score (P = 0.013) were higher and the VAS score (P = 0.011) was lower on the 77th day. Both Miao crossbow needle therapy and acupuncture reduced the WOMAC score. Miao crossbow needle therapy can be an alternative method for treating patients with knee osteoarthritis. ChiCTR, ChiCTR-INR-16008032. Registered on 12 March 2016.


Effect of Palonosetron on Physical Symptoms of Surgical Patients: A Systematic Review and Meta-Analysis.
Postoperative nausea and vomiting (PONV) is a typical and unpleasant physical symptom that occurs in patients after surgery, and it may be one of the most challenging elements of the recovery process. PONV can be caused by a variety of factors, including surgery itself, anesthesia, or medications. Palonosetron is a medication that is now licensed by the Food and Drug Administration for the treatment of this ailment. The efficacy of palonosetron in reducing physical symptoms in patients following surgery was investigated in this meta-analysis and comprehensive review. Following a quick search of databases such as CENTRAL, EMBASE, CINAHL, Google Scholar, the Science quotation index's Web site, the United States clinical trial check-in, the United Kingdom clinical trial check-in, the New Zealand clinical trial check-in, and the Australia check-in, as well as outlines of major anesthesia meetings held in the previous five years, we were able to get a good start on our research. Growing adults who had surgery and were given other drugs were compared to individuals who did get palonosetron in randomized controlled trials. A total of 8324 participants were recruited in 10 different clinical studies. It has been shown that palonosetron may significantly reduce the 24-hour PONV incidence and 95% confidence interval (CI) 0.41-0.86. When comparing the 6-hour and 48-hour time periods, the incidences of experiencing PONV were neither statistically different (RR: 0.82, 95% confidence interval: 0.61-1.09) or considerably different (RR: 0.60, 95% confidence interval: 0.33-1.10). Following in a similar vein, there was no significant difference between the groups in the occurrence of PONV after 48 hours (RR: 0.82, 95 percent CI: 0.59-1.14). The most often reported side effects of the medicine were headaches and dizziness, which were the most common. Regardless of the drug used, the difference in adverse reactions was not statistically significant. When it comes to the prevention of early PONV, it has been shown that palonosetron is more effective than other medications. Palonosetron, on the other hand, has been demonstrated to be more effective than other medications in preventing vomiting after laparoscopic surgery.


The influence of insulin, cAMP and the calcium ionophore X 537 A on the growth of the cartilage analagen of limb buds in vitro.
Limb buds of 11-day-old mouse embryos were cultured for 6 days with insulin, dibutyryl cAMP and X 537 A. The cartilage anlage was reduced by insulin and enlarged by dibutyryl cAMP and X 537 A. The effects are due to changes in the amount of intercellular substance.


Near-surface termination of upward-propagating strike-slip ruptures on the Yangsan Fault, Korea.
We present a new example of the termination of strike-slip paleoearthquake ruptures in near-surface regions on the Yangsan Fault, Korea, based on multi-scale structural observations. Paleoearthquake ruptures occur mostly along the boundary between the inherited fault core and damage zone (N10-20°E/> 75°SE). The ruptures propagated upward to the shallow subsurface along a < 3-cm-wide specific slip zone with dextral-slip sense, along which the deformation mechanism is characterized mainly by granular flow in near-surface region. The ruptures either reach the surface or are terminated in unconsolidated sediment below the surface. In the latter case, the rupture splays show westward bifurcation, and their geometry and kinematics show a change to NNW-strike with low-angle dip and dextral-reverse oblique-slip sense in the strata. We suggest that the upward termination of the contractional strike-slip ruptures is controlled by the inherited fault geometry that is unfavorable with respect to the stress field (ENE-WSW σHmax) at basement depths in terms of movement on the fault, and the lack of extension of the fault into shallow subsurface; a depth-dependent change in stress from σHmax > σv > σHmin to σHmax > σHmin > σv at depth of a ~ 200 m; and the physical properties of unconsolidated sediment, which have low inter-granular cohesion, resulting in distributed deformation.


[Transthyretin amyloid cardiomyopathy].
Transthyretin amyloid cardiomyopathy (ATTR-CM) is an underdiagnosed cause of heart failure and arrhythmia. This differential diagnosis should particularly be considered in older patients with left ventricular hypertrophy (LVH) who are also suffering from heart failure with preserved ejection fraction (HFpEF) or aortic valve stenosis. ATTR-CM is caused either by a genetic variation or by aging processes. The extracellular accumulation of amyloid fibrils in the heart causes a restrictive cardiomyopathy, which leads to typical heart failure symptoms as well as cardiac conduction and repolarization disturbances. Extracardiac problems such as a carpal tunnel syndrome can also be indicative for ATTR-CM. The disease can be diagnosed either by a myocardial biopsy or alternatively by a positive bone scintigraphy with the simultaneous exclusion of monoclonal proteins in blood and urine. Besides a symptomatic treatment, the transthyretin (TTR) stabilizer tafamidis is now available, which can significantly delay the disease progress. In the coming years, the approval of further drugs for the treatment of ATTR-CM is to be expected.


Hand washing with soap and water together with behavioural recommendations prevents infections in common work environment: an open cluster-randomized trial.
Hand hygiene is considered as an important means of infection control. We explored whether guided hand hygiene together with transmission-limiting behaviour reduces infection episodes and lost days of work in a common work environment in an open cluster-randomized 3-arm intervention trial. A total of 21 clusters (683 persons) were randomized to implement hand hygiene with soap and water (257 persons), with alcohol-based hand rub (202 persons), or to serve as a control (224 persons). Participants in both intervention arms also received standardized instructions on how to limit the transmission of infections. The intervention period (16 months) included the emergence of the 2009 influenza pandemic and the subsequent national hand hygiene campaign influencing also the control arm. In the total follow-up period there was a 6.7% reduction of infection episodes in the soap-and water arm (p = 0.04). Before the onset of the anti-pandemic campaign, a statistically significant (p = 0.002) difference in the mean occurrence of infection episodes was observed between the control (6.0 per year) and the soap-and-water arm (5.0 per year) but not between the control and the alcohol-rub arm (5.6 per year). Neither intervention had a decreasing effect on absence from work. We conclude that intensified hand hygiene using water and soap together with behavioural recommendations can reduce the occurrence of self-reported acute illnesses in common work environment. Surprisingly, the occurrence of reported sick leaves also increased in the soap-and water-arm. ClinicalTrials.gov: NCT00981877 The Finnish Work Environment Fund and the National Institute for Health and Welfare.


Impact of sleep on chronobiology of micturition among healthy older adults.
Nocturia (waking to void) is prevalent among older adults. Disruption of the well-described circadian rhythm in urine production with higher nighttime urine output is its most common cause. In young adults, their circadian rhythm is modulated by the 24-h secretory pattern of hormones that regulate salt and water excretion, including antidiuretic hormone (ADH), renin, angiotensin, aldosterone, and atrial natriuretic peptide (ANP). The pattern of hormone secretion is less clear in older adults. We investigated the effect of sleep on the 24-h secretion of these hormones in healthy older adults. Thirteen participants aged ≥65 yr old underwent two 24-h protocols at a clinical research center 6 wk apart. The first used a habitual wake-sleep protocol, and the second used a constant routine protocol that removed the influence of sleep, posture, and diet. To assess hormonal rhythms, plasma was collected at 8:00 am, 12:00 pm, 4:00 pm, and every 30 min from 7:00 pm to 7:00 am. A mixed-effects regression model was used to compare subject-specific and mean trajectories of hormone secretion under the two conditions. ADH, aldosterone, and ANP showed a diurnal rhythm that peaked during sleep in the wake-sleep protocol. These nighttime elevations were significantly attenuated within subjects during the constant routine. We conclude that sleep has a masking effect on circadian rhythm amplitude of ADH, aldosterone, and ANP: the amplitude of each is increased in the presence of sleep and reduced in the absence of sleep. Disrupted sleep could potentially alter nighttime urine output in healthy older adults via this mechanism.NEW & NOTEWORTHY Nocturia (waking to void) is the most common cause of sleep interruption among older adults, and increased nighttime urine production is its primary etiology. We showed that in healthy older adults sleep affects the 24-h secretory rhythm of hormones that regulate salt-water balance, which potentially alters nighttime urine output. Further studies are needed to elucidate the impact of chronic insomnia on the secretory rhythms of these hormones.


COVID-19 Time Capsule 2: DATE: July 23, 2020.
This Editorial is the second of COVID-19 Time Capsule. The first was dated on March 15, 2020. Time Capsule 2 is for July 23, 2020, but will publish in January/February 2021. Consider them "historical" documents-snapshots in time.


STRScan: targeted profiling of short tandem repeats in whole-genome sequencing data.
Short tandem repeats (STRs) are found in many prokaryotic and eukaryotic genomes, and are commonly used as genetic markers, in particular for identity and parental testing in DNA forensics. The unstable expansion of some STRs was associated with various genetic disorders (e.g., the Huntington disease), and thus was used in genetic testing for screening individuals at high risk. Traditional STR analyses were based on the PCR amplification of STR loci followed by gel electrophoresis. With the availability of massive whole genome sequencing data, it becomes practical to mine STR profiles in silico from genome sequences. Software tools such as lobSTR and STR-FM have been developed to address these demands, which are, however, built upon whole genome reads mapping tools, and thus may not be sensitive enough. In this paper, we present a standalone software tool STRScan that uses a greedy algorithm for targeted STR profiling in next-generation sequencing (NGS) data. STRScan was tested on the whole genome sequencing data from Venter genome sequencing and 1000 Genomes Project. The results showed that STRScan can profile 20% more STRs in the target set that are missed by lobSTR. STRScan is particularly useful for the NGS-based targeted STR profiling, e.g., in genetic and human identity testing. STRScan is available as open-source software at http://darwin.informatics.indiana.edu/str/ .


Cell death in osteoarthritis.
To date, most studies examining cell death during the development of osteoarthritis (OA) have focused on death of chondrocytes and have primarily examined advanced stages of the disease. Very good evidence suggests that chondrocyte death does occur at some point in the pathogenesis of OA and that it can be due to apoptosis, necrosis, or some combination of the two. Chondrocyte death can be induced by mechanical injury, loss of extracellular matrix, loss of growth factors, or excessive levels of reactive oxygen species. Although therapy specifically targeting cell death in human OA has not been reported, preclinical studies in animal models have provided early evidence that inhibition of caspases might slow OA-like changes in articular cartilage. Because of potential unwanted side effects from agents systemically inhibiting cell death, treatments specifically targeting cell death in OA will likely need to be delivered locally and in a manner that prevents systemic absorption. Inhibition of cell death in OA likely will not be a sole therapeutic target but rather a desired effect of interventions designed to reverse the catabolic-anabolic imbalance occurring in OA joint tissues.


A multicenter, randomized, open-label, pharmacokinetics and safety study of pantoprazole tablets in children and adolescents aged 6 through 16 years with gastroesophageal reflux disease.
Children with gastroesophageal reflux disease (GERD) may benefit from gastric acid suppression with proton pump inhibitors such as pantoprazole. Effective treatment with pantoprazole requires correct dosing and understanding of the drug's kinetic profile in children. The aim of these studies was to characterize the pharmacokinetic (PK) profile of single and multiple doses of pantoprazole delayed-release tablets in pediatric patients with GERD aged 6 to 11 years (study 1) and 12 to 16 years (study 2). Patients were randomly assigned to receive pantoprazole 20 or 40 mg once daily. Plasma pantoprazole concentrations were obtained at intervals through 12 hours after the single dose and at 2 and 4 hours after multiple doses for PK evaluation. PK parameters were derived by standard noncompartmental methods and examined as a function of both drug dose and patient age. Safety was also monitored. Pantoprazole PK was dose independent (when dose normalized) and similar to PK reported from adult studies. There was no evidence of accumulation with multiple dosing or reports of serious drug-associated adverse events. In children aged 6 to 16 years with GERD, currently available pantoprazole delayed-release tablets can be used to provide systemic exposure similar to that in adults.


Cytotoxicity of zinc in vitro.
The effect of zinc ions on B16 mouse melanoma lines, HeLa cells and I-221 epithelial cells was investigated in vitro in order to ascertain whether sensitivity to Zn2+ is a general feature of cells in vitro and in an attempt to elucidate the mechanism(s) of zinc cytotoxicity. The proliferation of B16, HeLa and I-221 cell lines was inhibited by 1.25 x 10(-4), 1.50 x 10(-4) and 1.50 x 10(-4) mol/l Zn2+, respectively. The free radical scavengers, methimazole and ethanol, did not suppress the toxicity of Zn2+, neither did superoxide dismutase or catalase. The addition of the chelating agent EDTA reduced the zinc cytotoxicity. It was possible to suppress the cytotoxicity of zinc by increasing the concentration of either Fe2+ or Ca2+ but not Mg2+, which suggests that a prerequisite for the toxic action of zinc is entry into cells using channels that are shared with iron or calcium. This view was supported by experiments in which transferrin intensified the cytotoxic action of zinc in serum-free medium. Another agent facilitating zinc transport, prostaglandin E2, inhibited the proliferation of the B16 melanoma cell line. There were no conspicuous differences in zinc toxicity to pigmented and unpigmented cells. The toxic effect of zinc in the cell systems studied exceeded that of iron, copper, manganese and cobalt in the same concentration range. In vitro, Zn2+ should be regarded as a dangerous cation.


'Small communities, large oversight': The impact of recent legislative changes concerning supervised consumption services on small communities in Ontario, Canada.
The opioid epidemic continues to exert a large toll on public health across Canada, with the province of Ontario specifically experiencing exceptionally high opioid-related hospitalizations and overdoses in 2019. As a key evidence-based public health intervention to reduce these harms, over the past few years supervised consumption services (SCS), including temporary and flexible unsanctioned models, have been implemented in select cities across the province. However, in March 2019, the newly elected Ontario provincial government replaced SCS regulations with a 'streamlined' model that introduced additional bureaucratic and administrative requirements for application approval and funding, while placing a cap on the number of sites allowed. Only a select few sites were approved, despite the ability of numerous cities across the province to demonstrate a need for SCS in their community. Importantly, many small communities throughout Ontario have been experiencing opioid-related harms at rates which far outpace those of larger cities. Several of these communities have been actively working towards securing SCS sites, but have been unable to do so as they do not have the resources to meet stringent application requirements. These regulatory changes therefore hinder the feasibility of small communities to implement necessary and life-saving SCS, which will affect lives. This is a major oversight.


Simple and rapid detection of L-Dopa decarboxylase activity using gold nanoparticles.
We developed a new detection method for L-Dopa decarboxylase (DDC) activity using gold nanoparticles (AuNPs). When L-3,4-dihydroxyphenylalanine (L-Dopa) is decarboxylated to dopamine (DA) by DDC, DA induces the aggregation of AuNPs, and the color of the AuNPs changes from red to blue.


Controlled charged amino acids of Ti-binding peptide for surfactant-free selective adsorption.
The adsorption mechanism of titanium-binding peptide (TBP) on metal oxide substrates was investigated by evaluating the adsorption behavior of ferritins with various alanine-substituted TBPs. Results revealed that (a) a positively charged amino acid, lysine (K) or arginine (R), in TBP can anchor ferritin to negative zeta-potential substrates, (b) the adsorption force of K is stronger than R, and (c) local electrostatic interactions and flexibility of TBP directly affect adsorption. Based on these findings, selective ferritin adsorption on SiO2 with TiOX patterned surfaces in a surfactant-free condition was demonstrated. Alanine-substituted TBP with one positively charged amino acid (K) and one negatively charged amino acid (D), achieved ferritin-selective adsorption without a surfactant. The importance of controlled electrostatic forces between TBP and a substrate for selective adsorption without a surfactant was clearly demonstrated.


Specific roles for the PI3K and the MEK-ERK pathway in IGF-1-stimulated chemotaxis, VEGF secretion and proliferation of multiple myeloma cells: study in the 5T33MM model.
Insulin-like growth factor-1 (IGF-1) has been described as an important factor in proliferation, cell survival and migration of multiple myeloma (MM) cells. Angiogenesis correlates with development and prognosis of the MM disease. Vascular endothelial growth factor (VEGF) is one of the prominent factors involved in this process. The different functions of IGF-1 were investigated in the 5TMM mouse model with emphasis on proliferation, migration and VEGF secretion, and the signalling pathways involved. Western Blot analysis revealed that ERK1/2 and Akt (PKB) were activated after IGF-1 stimulation. The activation of ERK1/2 was reduced by the PI3K inhibitor Wortmannin, implying that the PI3K pathway is involved in its activation. Insulin-like growth factor-1 induced an increase in DNA synthesis in MM cells, which was mediated by a PI3K/Akt-MEK/ERK pathway. Insulin-like growth factor-1 enhanced F-actin assembly and this process was only PI3K mediated. Stimulation by IGF-1 of VEGF production was reduced by PD98059, indicating that only the MEK-ERK pathway is involved in IGF-1-stimulated VEGF production. In conclusion, IGF-1 mediates its multiple effects on MM cells through different signal transduction pathways. In the future, we can study the potential in vivo effects of IGF-1 inhibition on tumour growth and angiogenesis in MM.


Temporary cardiac pacing for fatal arrhythmia in living-donor liver transplantation: three case reports.
Cardiac pacing often turns out to be the only effective treatment of severe, life-threatening arrhythmias. We performed 77 living-donor liver transplantations (LDLT) from 1999 to 2007. In these cases, three recipients experienced fatal arrhythmia and required temporary cardiac pacing during the perioperative period. The first case was a 68-year-old woman diagnosed with liver cirrhosis and hepatocellular carcinoma (HCC). Her Model for End-Stage Liver Disease (MELD) score was 34. We performed LDLT using a right lobe graft. She showed complete atrioventricular block with cardiac arrest at postoperative day (POD) 42 after a bacterial infection. We performed a resuscitation and instituted temporary cardiac pacing. However, she was dead at POD 43. Pathologic findings at autopsy showed a diffuse myocardial abscess, which caused the fatal arrhythmia. The second case was a 58-year-old man diagnosed with HCC and liver cirrhosis; his MELD score was 9. We performed LDLT using a right lobe graft. He showed atrial fibrillation after septic shock. He also showed sinus bradycardia with a cardiac arrest at POD 10. We performed resuscitation and emergent temporary pacing. He recovered and was alive without recurrence of arrhythmia or infection. The third case was a 58-year-old woman diagnosed with multiple HCC. During preoperative regular check-up, she was diagnosed to have cardiac hypertrophy and was started on beta-blockers as treatment for cardiac hypertrophy. However, severe bradycardia necessitated temporary cardiac pacing. LDLT was performed safely after implantation of a pacemaker. Early use of temporary cardiac pacing for severe arrhythmias may be effective to maintain the hemodynamic state in LDLT.


Abnormal Brain Protein Abundance and Cross-tissue mRNA Expression in Amyotrophic Lateral Sclerosis.
Due to the limitations of the present risk genes in understanding the etiology of amyotrophic lateral sclerosis (ALS), it is necessary to find additional causative genes utilizing novel approaches. In this study, we conducted a two-stage proteome-wide association study (PWAS) using ALS genome-wide association study (GWAS) data (N = 152,268) and two distinct human brain protein quantitative trait loci (pQTL) datasets (ROSMAP N = 376 and Banner N = 152) to identify ALS risk genes and prioritized candidate genes with Mendelian randomization (MR) and Bayesian colocalization analysis. Next, we verified the aberrant expression of risk genes in multiple tissues, including lower motor neurons, skeletal muscle, and whole blood. Six ALS risk genes (SCFD1, SARM1, TMEM175, BCS1L, WIPI2, and DHRS11) were found during the PWAS discovery phase, and SARM1 and BCS1L were confirmed during the validation phase. The following MR (p = 2.10 × 10-7) and Bayesian colocalization analysis (ROSMAP PP4 = 0.999, Banner PP4 = 0.999) confirmed the causal association between SARM1 and ALS. Further differential expression analysis revealed that SARM1 was markedly downregulated in lower motor neurons (p = 7.64 × 10-3), skeletal muscle (p = 9.34 × 10-3), and whole blood (p = 1.94 × 10-3). Our findings identified some promising protein candidates for future investigation as therapeutic targets. The dysregulation of SARM1 in multiple tissues provides a new way to explain ALS pathology.