Random mobility and spatial structure often enhance cooperation.
The effects of an unconditional move rule in the spatial Prisoner's Dilemma, Snowdrift and Stag Hunt games are studied. Spatial structure by itself is known to modify the outcome of many games when compared with a randomly mixed population, sometimes promoting, sometimes inhibiting cooperation. Here we show that random dilution and mobility may suppress the inhibiting factors of the spatial structure in the Snowdrift game, while enhancing the already larger cooperation found in the Prisoner's dilemma and Stag Hunt games.


Translin restricts the growth of pubertal mammary epithelial cells estrogen-independently in mice.
Translin, a ubiquitous RNA/DNA-binding protein that forms a hetero-octamer together with Translin-associated factor X (TRAX), possesses endoribonuclease activity and plays a physiological role in restricting the size and differentiation of mesenchymal precursor cells. However, the precise role of Translin in epithelial cells remains unclear. Here, we show evidence that Translin restricts the growth of pubertal mammary epithelial cells. The mammary epithelia of Translin-null females exhibited retarded growth before puberty, but highly enhanced growth and DNA synthesis with increased ramification after the onset of puberty. Primary cultures of Translin-null mammary epithelial cells showed augmented DNA synthesis in a ligand-independent and ligand-enhanced manner. Translin-null ovariectomized mice implanted with slow-release estrogen pellets showed enhanced length and ramification of the mammary glands. Mammary epithelial growth was also observed in ovariectomized Translin-null mice implanted with placebo pellets. Luciferase reporter assays using embryonic fibroblasts from Translin-null mice showed unaltered estrogen receptor α function. These results indicate that Translin plays a physiological role in restricting intrinsic growth, beyond mesenchymal cells, of pubertal mammary epithelial cells.


Binary blended co-delivery nanoparticles with the characteristics of precise pH-responsive acting on tumor microenvironment.
Although combined chemotherapy had achieved the ideal efficacy in clinical anti-cancer therapeutic, the issues that need to be addressed are non-targeting and toxic-side effects of small molecule chemical drug (SMCD). In this study, we designed and prepared a novel binary blended co-delivered nanoparticles (BBCD NPs) with pH-responsive feature on tumor microenvironment. The BBCD NPs consists of two kind of drug-loaded NPs, in one of which carboxymethyl chitosan (CMC) and Poly (lactic-co-glycolic acid) (PLGA) were chosen as delivery carrier to load anti-cancer drug vincristine (VCR), named CMC-PLGA-VCR NPs (or CPNPVCR); and in the other of which methoxy poly(ethylene glycol)-poly(β-amino ester) (mPEG-PAE) were chosen as delivery carrier to load anti-fibrotic drug pirfenidone (PFD), named mPEG-PAE-PFD NPs (or PPNPPFD). Then, the two types of NPs (CPNPVCR and PPNPPFD) were physically mixed in mass ratios to form BBCD NPs, which was named CPNPVCR&PPNPPFD. CPNPVCR&PPNPPFD had good encapsulation efficiency and loading capacity, and the particle size distribution was uniform. In cytotoxicity experiments and non-contact co-culture studies in vitro, the model drugs loaded in CPNPVCR&PPNPPFD could respectively target cancer cell and cancer associated fibroblast (CAF) owing to the precise pH-sensitive drug release in the pharmacological targets and show stronger synergism than that of the combined treatment of two free drugs. As a modularity and assemble ability feature in design, BBCD NPs would have the advantages on the terms of concise on preparation process, controllable on quality standard, stable in natural environment storage. The research results can provide scientific evidence for the further development of a novel drug co-delivery system with multi-type cell targets.


Role of Radiation in the Era of Effective Systemic Therapy for Melanoma.
The role of radiation therapy in melanoma has evolved over the last few decades. There has been a dramatic improvement in radiation delivery with the introduction of intensity-modulated radiation therapy, image-guided radiation therapy, stereotactic radiosurgery, and stereotactic body radiation therapy/stereotactic radiation therapy. More recently, with the introduction of immunotherapy in various malignancies, including melanoma, the role of radiation therapy is being reevaluated. This article describes the evolution of the role of radiation therapy from nonimmunotherapy to the era of immunotherapy.


Peptide Regulation of Skin Fibroblast Functions during Their Aging In Vitro.
The effect peptides KE, KED, AED and AEDG on proliferation (Ki-67), regeneration and aging (CD98hc), apoptosis (caspase-3), and extracellular matrix remodeling (MMP-9) in skin fibroblasts during their aging in culture were studied by immunofluorescent confocal microscopy. All studied peptides inhibited MMP-9 synthesis that increases during aging of skin fibroblasts and enhanced the expression of Ki-67 and CD98hc that are less intensively synthesized during cell aging. Peptides AED and AEDG suppressed caspase-dependent apoptosis that increases during aging of cell cultures.


Lepidodactylus lugubris (Duméril & Bibron 1836) (Reptilia: Gekkonidae), an introduced lizard new for Brazil, with remarks on and correction of its distribution in the New World.
Recently seven specimens of the gecko Lepidodactylus lugubris were collected or observed in Belém, Pará, northern Brazil. This is the first vouchered occurrence in Brazil of a widely dispersed (Pacific area) and invasive species (much of the Pacific, parts of northern South America and southern Central America and Florida, U.S.A.). In Suriname the species has already spread into the interior. The distribution of the species is corrected and the history of its introduction in the New World is reconstructed, with an estimation of the state of invasiveness for each country. Some possible routes of introduction are discussed.


Changes in public perceptions and experiences of the Australian health-care system: A decade of change.
The views and experiences of the Australian public are an important barometer of the health system. This study provides key findings about the changing views held by Australians over time regarding their individual experiences and perceptions of the overall performance of the health system. A population-based online survey was conducted in 2018 (N = 1024). Participants were recruited through market research panels. The results were compared with previous Australian population survey data sets from 2008 (N = 1146), 2010 (N = 1201) and 2012 (N = 1200), each of which used different population samples. The survey included questions consistent with previous surveys regarding self-reported health status, and questions about use, opinions and experiences of the health system. Overall, there has been a shift in views from 2008 to 2018, with a higher proportion of respondents now viewing the Australian health-care system more positively (X2 (2, N = 4543) = 96.59, P < .001). In 2018, areas for attention continued to include the following: the need for more doctors, nurses and other health workers (29.0%); lower costs for care or Orion medicines (27.8%); more access to care (13.1%); and enhancements in residential aged care (17.3% rated these services as 'bad' or 'very bad'). This research suggests that Australians' perceptions of their health-care system have significantly improved over the last decade; however, concerns have emerged over access to medicines, inadequate workforce capacity and the quality of aged care facilities. Our study highlights the value of periodically conducting public sentiment surveys to identify potential emerging health system problems.


Distinct Characteristics and Metastatic Behaviors of Late Recurrence in Patients With Hormone Receptor-positive/Human Epidermal Growth Factor Receptor 2-negative Breast Cancer: A Single Institute Experience of More Than 10 Years.
Patients with hormone receptor-positive and human epidermal growth factor receptor 2-negative (HR+/HER2-) breast cancer have a constant risk of relapse over time. Nearly one-half of the recurrences occur more than 5 years after diagnosis, described as late recurrence, but little is known about late recurrence. We reviewed the clinical data of 1941 patients with HR+/HER2- breast cancer who had operations in the Cancer Institute and Hospital, Chinese Academy of Medical Sciences, during 2003 to 2009, and found 390 relapsed cases. Among them, 281 patients were early recurrence, and 109 were late recurrence. In the late recurrence group, patients with ≤ 3 lymph node metastases, double HR+ (estrogen receptor-positive/progesterone receptor-positive) were more common (72.48% vs. 55.52%; P = .005; 82.57% vs. 71.89%; P = .029, respectively) when compared with the early recurrence group. The lung seemed to be a preferential site of late recurrence. Although visceral disease and multi-organ metastases were more frequent in the late recurrence group, survival after recurrence was significantly longer than that in the early recurrence group (52 vs. 40 months; hazard ratio, 1.508; 95% confidence interval, 1.142-1.992; P = .003). Moreover, progression-free survival of first-line treatment was an independent prognostic factor of survival after recurrence in the late recurrence group. Late recurrence differed from early recurrence in many ways in HR+/HER2- breast cancer, and its prognosis was much better. The lung may be a preferential site of late recurrence. More attention should be paid to late recurrence itself.


Maternal approach behaviors toward neonatal calls are impaired by mother's experiences of raising pups with a risk gene variant for autism.
How the intrinsic sequence structure of neonatal mouse pup ultrasonic vocalization (USV) and maternal experiences determine maternal behaviors in mice is poorly understood. Our previous work showed that pups with a Tbx1 heterozygous (HT) mutation, a genetic risk for autism spectrum disorder (ASD), emit altered call sequences that do not induce maternal approach behaviors in C57BL6/J mothers. Here, we tested how maternal approach behaviors induced by wild-type and HT USVs are influenced by the mother's experience in raising pups of these two genotypes. The results showed that wild-type USVs were effective in inducing maternal approach behaviors when mothers raised wild-type but not HT pups. The USVs of HT pups were ineffective regardless of whether mothers raised HT or wild-type pups. However, the sequence structure of pup USVs had no effect on the general, non-directional incentive motivation of maternal behaviors. Our data show how the mother's experience with a pup with a genetic risk for ASD alters the intrinsic incentive values of USV sequences in maternal approach behaviors.


Cyanamide mediated syntheses under plausible primitive earth conditions. II. The polymerization of deoxythymidine 5'-triphosphate.
When an aqueous solution (pH 7.0) of 3H deoxythymidine 5'-triphosphate, deoxythymidine 5'-phosphate, 4-amino-5-imidazolecarboxamide, cyanamide and ammonium chloride was dried and heated at 60 degrees C for 18 h, oligomers were obtained in a yield of approximately 80%. After the chemical degradation of any pyrophosphate bonds present in these oligomers, linear polynucleotides of up to 7-8 units in length were isolated by DEAE cellulose column chromatography and identified by enzymatic digestion procedures. The di- and trinucleotide fractions were degraded 87% and 100% by snake venom phosphodiesterase and 39% and 9% by spleen phosphodiesterase. This synthesis of deoxythymidine oligonucleotides was conducted under potentially prebiotic conditions and may offer a possible method for the synthesis of deoxyoligonucleotides on the primitive Earth.


An unusual case of complete heart block with triplet pregnancy.
A young primigravida presented at 36 weeks of gestation with complete heart block and triplet pregnancy. She underwent a lower segment cesarean section, and was managed successfully. The patient remained asymptomatic and did not require antiarrhythmic drugs or pacing.


Functional screening of serine protease inhibitors in the medical leech Hirudo medicinalis monitored by intensity fading MALDI-TOF MS.
The blood-feeding invertebrates are a rich biological source of drugs and lead compounds to treat cardiovascular diseases because they have evolved highly efficient mechanisms to feed on their hosts by blocking blood coagulation. In this work, we focused our attention on the leech Hirudo medicinalis. We performed, by "intensity fading" MALDI-TOF mass spectrometry, a comprehensive detection and functional analysis of pre-existent peptides and small proteins with the capability of binding to trypsin-like proteases related to blood coagulation. Combining "intensity fading MS" and off-line LC prefractionation allowed us to detect more than 75 molecules present in the leech extract that interact specifically with a trypsin-like protease over a sample profile of nearly 2,000 different peptides/proteins in the 2-20-kDa range. Moreover we resolved 232 individual components from the complex mixture, 13 of which have high sequence homology with previously described serine protease inhibitors. Our findings indicate that such extracts are much more complex than expected. Additionally, intensity fading MS, when complemented with LC separation strategies, seems to be a useful tool to investigate complex biological samples, establishing a new bridge between profiling, functional peptidomics, and subsequent drug discovery.


Chemical factors governing the state of saturation towards brushite and whewellite in urine of calcium stone formers.
Variations of urinary pH and concentrations of calcium, phosphate, oxalate, magnesium and citrate have been produced by 4 different diets given to 19 idiopathic calcium stone formers. The state of saturation towards whewellite and brushite was directly measured in the 76 urine samples by equilibration with the corresponding salts and was compared to chemical constituents by regression analyses. The state of saturation towards calcium oxalate monohydrate was significantly governed only by the urinary oxalate concentration, and a soluble oxalate fraction not contributing to calcium oxalate chelation was demonstrated. The state of saturation towards brushite was exclusively determined by urinary calcium and pH, the latter below 5.5 showing a high influence on brushite solubility.


Long-term monitoring of the diabetic dog and cat. Clinical signs, serial blood glucose determinations, urine glucose, and glycated blood proteins.
Management of diabetic dogs or cats requires the use of all available monitoring technology (Fig. 6). First, one should ask questions about the clinical control of DM. Are the clinical signs of DM resolved, and is the owner satisfied with insulin therapy? Other important questions would include: Is the dog or cat developing long-term complications of diabetes such as neuropathies or cataracts? Is body weight remaining stable? Is the dog or cat showing any signs of hypoglycemia? One should determine if the blood glucose curves are close to ideal for the type of insulin being administered. Urine glucose and ketones should be negative or trace as assessed by the at-home monitoring by the owner. In the problem diabetic, long-term glucose control can be assessed by serum fructosamine or glycosylated hemoglobin determinations. Regulation of the diabetic patient is accomplished when the owner is satisfied with the therapy and when the serum glucose monitoring parameters are acceptable.


Ultrafast and high-throughput quantitative analysis of carbamazepine in human plasma by direct analysis in real time tandem mass spectrometry coupled with solid phase extraction to eliminate matrix effects.
Therapeutic drug monitoring of carbamazepine is necessary for its clinical application with the purpose to reach therapeutic concentration and reduce the risk of concentration-dependent toxicity. An ultrafast analytical assay for quantification of carbamazepine in human plasma was developed and validated based on direct analysis in real time tandem mass spectrometry (DART-MS/MS). After reversed phase solid phase extraction with Waters Oasis HLB, carbamazepine and internal standard carbamazepine-D2N15 were monitored by positive ion mode followed by multiple reaction monitoring (MRM) of the transitions at m/z 237.1→194.0 and 240.1→196.2, respectively. The DART-MS/MS method is ultrafast and high-throughput and the analytical time for each sample is only 0.4 min. The assay was linear in the concentration range 0.50-30 μg/mL and intra- and inter-day accuracies were within ± 15% and trueness were < 13.9% at all concentrations. The method was successfully applied to therapeutic drug monitoring of carbamazepine in human plasma.


Fatal Status Epilepticus in Dravet Syndrome.
Dravet Syndrome (DS) is burdened by high epilepsy-related premature mortality due to status epilepticus (SE). We surveyed centres within Europe through the Dravet Italia Onlus and EpiCARE network (European Reference Network for Rare and Complex Epilepsies). We collated responses on seven DS SCN1A+ patients who died following refractory SE (mean age 6.9 year, range 1.3-23.4 year); six were on valproate, clobazam, and stiripentol. All patients had previous SE. Fatal SE was always triggered by fever: either respiratory infection or one case of hexavalent vaccination. SE lasted between 80 min and 9 h and all patients received IV benzodiazepines. Four patients died during or within hours of SE; in three patients, SE was followed by coma with death occurring after 13-60 days. Our survey supports the hypothesis that unresponsive fever is a core characteristic feature of acute encephalopathy. We highlight the need for management protocols for prolonged seizures and SE in DS.


Molecular characterization of the RNA S segment of nephropathia epidemica virus strain Hällnäs B1.
The S segment RNA of nephropathia epidemica virus (NEV) strain Hällnäs B1 was isolated by molecular cloning of the corresponding cDNA. The RNA is 1785 nucleotides long with the 3' and 5' termini being complementary for 23 bases. The viral messenger-sense RNA contains one major open reading frame (ORF) with a coding capacity of 433 amino acids encoding a 49-kDa polypeptide. Compared to the Hantaan S segment cDNA sequence there is a nucleotide homology of 60 and 61% at the amino acid level. Many of the amino acid differences are conservative exchanges. The C-termini of the NEV and Hantaan nucleocapsid proteins are nearly identical and the hydrophilicity profiles are very similar. In contrast, the following differences are significant: The calculated isoelectric points of the NEV and Hantaan nucleocapsid proteins are 5.6 and 6.7, respectively. The most prominent antigenic determinants predicted by the hydrophilicity profiles are located close to the C-terminus of NEV and close to the N-terminus of Hantaan virus nucleocapsid polypeptides.


Low incidence of chest wall pain with a risk-adapted lung stereotactic body radiation therapy approach using three or five fractions based on chest wall dosimetry.
To examine the frequency and potential of dose-volume predictors for chest wall (CW) toxicity (pain and/or rib fracture) for patients receiving lung stereotactic body radiotherapy (SBRT) using treatment planning methods to minimize CW dose and a risk-adapted fractionation scheme. We reviewed data from 72 treatment plans, from 69 lung SBRT patients with at least one year of follow-up or CW toxicity, who were treated at our center between 2010 and 2013. Treatment plans were optimized to reduce CW dose and patients received a risk-adapted fractionation of 18 Gy×3 fractions (54 Gy total) if the CW V30 was less than 30 mL or 10-12 Gy×5 fractions (50-60 Gy total) otherwise. The association between CW toxicity and patient characteristics, treatment parameters and dose metrics, including biologically equivalent dose, were analyzed using logistic regression. With a median follow-up of 20 months, 6 (8.3%) patients developed CW pain including three (4.2%) grade 1, two (2.8%) grade 2 and one (1.4%) grade 3. Five (6.9%) patients developed rib fractures, one of which was symptomatic. No significant associations between CW toxicity and patient and dosimetric variables were identified on univariate nor multivariate analysis. Optimization of treatment plans to reduce CW dose and a risk-adapted fractionation strategy of three or five fractions based on the CW V30 resulted in a low incidence of CW toxicity. Under these conditions, none of the patient characteristics or dose metrics we examined appeared to be predictive of CW pain.


Deconstructing SARS-CoV-2 neutralization: A modular molecular framework for computational design and comparison of antibodies and nanobodies targeting the spike RBD.
Since 2020 the COVID-19 pandemic has led scientists to search for strategies to predict the transmissibility and virulence of new severe acute respiratory syndrome coronavirus 2 variants based on the estimation of the affinity of the spike receptor binding domain (RBD) for the human angiotensin-converting enzyme 2 (ACE2) receptor and/or neutralizing antibodies. In this context, our lab developed a computational pipeline to quickly quantify the free energy of interaction at the spike RBD/ACE2 protein-protein interface, reflecting the incidence trend observed in the transmissibility/virulence of the investigated variants. In this new study, we used our pipeline to estimate the free energy of interaction between the RBD from 10 variants, and 14 antibodies (ab), or 5 nanobodies (nb), highlighting the RBD regions preferentially targeted by the investigated ab/nb. Our structural comparative analysis and interaction energy calculations allowed us to propose the most promising RBD regions to be targeted by future ab/nb to be designed by site-directed mutagenesis of existing high-affinity ab/nb, to increase their affinity for the target RBD region, for preventing spike-RBD/ACE2 interactions and virus entry in host cells. Furthermore, we evaluated the ability of the investigated ab/nb to simultaneously interact with the three RBD located on the surface of the trimeric spike protein, which can alternatively be in up- or down- (all-3-up-, all-3-down-, 1-up-/2-down-, 2-up-/1-down-) conformations.


Suicides among military conscripts between 1991-2007 in Finland--a descriptive replication study.
Suicide mortality in Finland particularly among young people remains high, compared with other European countries. Suicide is one of the leading causes of death among military conscripts in Finland. In this record-based study, we investigated characteristics of all 48 suicides among Finnish conscripts during the period 1991-2007. We collected and analyzed healthcare records during military service and medicolegal investigations (medicolegal autopsy) of suicide victims. Of the 48 suicides, 36 (75%) were completed during leave days. Shooting, hanging and jumping were the most common methods used; a service gun was used in four (14%) of the 29 shooting cases. One-third of the victims had been in contact with military healthcare during the week preceding their suicide, and two-thirds during the preceding month. When conscripts use healthcare services, suicidal thoughts should be actively asked of those conscripts with history of any risk factors for suicidal behavior.


Ion storage biases in the ion funnel trap of a Hybrid ion mobility spectrometer/time of flight mass spectrometer.
A detailed experimental characterization on the ion storage biases in an ion funnel trap, related to ion structure, charge state and RF voltage applied to the ion funnel trap, is reported by using both cytochrome C and ubiquitin samples. It was first observed experimentally that an unavoidable ion overflow would occur when the incoming ions exceeded the capacity of ion funnel trap. The conformers with extended structures would lose preferentially in the ion overflow process. Accordingly, a significant structural bias in the ion mobility spectrometry/time of flight mass spectrometry (IMS-TOF MS) spectrum was created, as the peak intensities for conformers with compact structures and extended structures would continuously increase and decrease, respectively, when the ion overflow time of the ion funnel trap was increased. Furthermore, the experimental results also showed that the effect of this ion structural bias was more significant when the RF voltage applied to the ion funnel trap was increased. In addition, an ion charge state bias in the ion funnel trap was also observed. The effect of the ion structural bias depends significantly on the specific charge state of the ions. For a given analyte, its lower charge state ions show a greater sensitivity to the ion structural bias than the higher charge state ones under the same ion funnel trap operating conditions. Therefore, it is extremely important to set a reasonable operation condition for the ion funnel trap to avoid ion storage biases in IMS-TOF MS.


In vitro activity of phospholipase A2 and of peptides from Crotalus durissus terrificus venom against amastigote and promastigote forms of Leishmania (L.) infantum chagasi.
American visceral leishmaniasis is caused by the intracellular parasite Leishmania (L.) infantum chagasi, and transmitted by the sand fly Lutzomyia longipalpis. Since treatment is based on classical chemotherapeutics with significant side effects, the search for new drugs remains the greatest global challenge. Thus, this in vitro study aimed to evaluate the leishmanicidal effect of Crotalus durissus terrificus venom fractions on promastigote and amastigote forms of Leishmania (L.) infantum chagasi. Phospholipase A2 (PLA2) and a pool of peptide fraction (<3 kDa) were purified from Crotalus venom. Furthermore, promastigotes and peritoneal macrophages of mice infected by amastigotes were exposed to serial dilutions of the PLA2 and peptides at intervals varying between 1.5625 μg/mL and 200 μg/mL. Both showed activity against promastigotes that varied according to the tested concentration and the time of incubation (24, 48 and 72 h). MTT assay for promastigotes showed IC50 of 52.07 μg/mL for PLA2 and 16.98 μg/mL for the peptide fraction of the venom. The cytotoxicity assessment in peritoneal macrophages showed IC50 of 98 μg/mL and 16.98 μg/mL for PLA2 and peptide by MTT assay, respectively. In peritoneal macrophages infected by Leishmania (L.) infantum chagasi amastigotes, the PLA2 stimulated growth of parasites, and at higher doses reduced growth by 23 %. The peptide fraction prevented 43 % of the intracellular parasite growth at a dose of 16.98 μg/mL, demonstrating the toxicity of this dose to macrophages. Both fractions stimulated H2O2 production by macrophages but only PLA2 was able to stimulate NO production. We have demonstrated the in vitro leishmanicidal activity of the PLA2 and peptide fraction of Crotalus venom. The results encourage further studies to describe the metabolic pathways involved in cell death, as well as the prospecting of molecules with antiparasitic activity present in the peptide fraction of Crotalus durissus terrificus venom.


A Screening Method to Identify Essential Yeast Genes for Responses Towards Spermine.
We exploited the yeast DAmP mutant collection to identify essential genes that play a role in polyamine resistance. Herein, we described in details the methodology to obtain these genes. This approach is applicable for screening many nontoxic and toxic drugs.


Perinatal cocaine intoxication.
This report describes a case in which cocaine abuse during pregnancy was related temporally to abruptio placentae, preterm labour and to rapid delivery. The neonate developed respiratory distress with features of persistent pulmonary hypertension of the newborn and also signs of neurological dysfunction. He was managed by supportive therapy and has been followed-up to nine months of age. Perinatal cocaine intoxication, either directly or indirectly, is discussed as a cause of his cardiorespiratory and neurological maladaptation. As cocaine abuse appears to be an escalating problem in Australia, increased public awareness of the dangers of both acute and long-term abuse of the drug is seen as a priority in preventive care.


Effects of cadmium exposure on expression and activity of P-glycoprotein in eastern oysters, Crassostrea virginica Gmelin.
Heavy metal pollution is a worldwide problem, and cadmium (Cd) is one of the most noxious pollutants in aquatic environments. We studied P-glycoprotein (P-gp) expression and function in control and Cd exposed (50microgL(-1) Cd, 30-40 days) oysters Crassostrea virginica as a possible mechanism of cell protection against Cd. Our data show that P-gp is expressed on cell membrane and in mitochondria of oyster gills and hepatopancreas. Inhibitor studies with verapamil, cyclosporine A and JS-2190 suggest that in the gills, mitochondrial P-gp pumps substrates from cytosol into the mitochondria, while cell membrane P-gp pumps substrates from cytosol out of the cell. Cd exposure resulted in a 2-2.5-fold increase in P-gp protein expression in cell membranes and a 3.5-7-fold increase in transport activity measured as the inhibitor-sensitive rhodamine B extrusion rate. In contrast, p-gp mRNA levels were similar in control and Cd-exposed oysters. No difference in P-gp protein expression was observed between mitochondria of control and Cd-exposed oysters but the apparent transport activity was higher in mitochondria from Cd-exposed oysters. Overall, a stronger increase in substrate transport activity in Cd-exposed oysters compared to a relatively weaker change in P-gp protein levels suggests that P-gp activity is post-translationally regulated. Our data show that direct determination of P-gp transport activity may be the best measure of the xenobiotic-resistant phenotype, whereas p-gp mRNA levels are not a good marker due to the likely involvement of multiple post-transcriptional regulatory steps. Cd exposure resulted in a significantly elevated rate of oxygen consumption of isolated oyster gills by 46%. Specific inhibitors of ATPase function of P-gp (cyclosporine A and JS-2190) had no significant effect on tissue oxygen consumption indicating that P-gp contribution to energy budget is negligible and supporting indirect estimates based on the ATP stoichiometry of substrate transport that also suggest low energy demand for P-gp function.


The role of Na(V)1.8 sodium channel in the maintenance of chronic inflammatory hypernociception.
We previously described an animal model of persistent inflammatory sensitization of nociceptors. In this model the hypernociception persists for more than 30 days after the cessation of 2 weeks of daily intraplantar treatment with prostaglandin E(2) (PGE(2)). The tetrodotoxin-resistant (TTX-R) voltage-gated sodium channel Na(V)1.8 is considered a characteristic of primary afferent nociceptive C fibers and plays an important role in acute hypernociception. In the present study, the relevance of the Na(V)1.8 channel was investigated in this model of persistent mechanical hypernociception in rats. In the PGE(2)-induced persistent hypernociception, but not in the single injection-induced acute hypernociception, the mRNA expression (RT-PCR) of Na(V)1.8 in dorsal root ganglia (DRG) was up-regulated. A similar increase of Na(V)1.8 mRNA was observed when DbcAMP was used to induce persistent hypernociception. Four daily intrathecal administrations of oligodeoxynucleotides (ODN) antisense against Na(V)1.8 decreased the mRNA encoding Na(V)1.8 in DRG. The intrathecal administration of ODN antisense prevented the PGE(2)-induced acute hypernociception and significantly reduced ongoing PGE(2)-induced persistent hypernociception. A parallel restoration of the persistent hypernociception and up-regulation of Na(V)1.8 mRNA was observed after the cessation of ODN antisense treatment. These results suggest the participation of Na(V)1.8 channels in the development and maintenance of chronic inflammatory hyperalgesia, and confirm their involvement in the acute inflammatory hypernociception.


Multicenter prospective study of the efficacy of stereomicroscopic on-site evaluation in endoscopic ultrasound-guided tissue acquisition in patients with pancreatic cancer.
In sample isolation processing by stereomicroscopy (SIPS), a technique used to assess the quality of specimens collected during endoscopic ultrasound (EUS)-guided tissue acquisition (EUS-TA), the cutoff value of stereomicroscopically visible white core (SVWC) (≥11 mm) indicates high diagnostic sensitivity. However, the procedure of SIPS is complicated and time-consuming. Therefore, we devised the stereomicroscopic on-site evaluation (SOSE), a new rapid assessment method that is simpler than SIPS and only determines if the SVWC cutoff value is attained. We aimed to examine the usefulness of SOSE in a multicenter, prospective setting. Seventy patients from multiple institutions with solid pancreatic masses suspected to be pancreatic cancer were included. EUS-TA was performed using a 22-gauge Franseen needle. SVWCs were measured on-site using stereomicroscopy. The primary outcome was the sensitivity of SVWC cutoff value in EUS-TA with SOSE. The total number of punctures was 214 and SOSE was performed on 150 punctures. The SVWC cutoff value collection rate was 100% per lesion, with 80% in the first pass, 79% in the second pass, and 78% per puncture in all passes. The median time taken to determine the SVWC cutoff value for SOSE was 47 s. The sensitivity of the SVWC cutoff value was 93.2% for histology and 96.6% for cytology + histology. The per-lesion accuracy of pathological diagnosis reached the highest level (98.6%) at the second puncture. SOSE showed high diagnostic sensitivity and may be a new rapid assessment method for the diagnosis of malignant pancreatic cancer using EUS-TA.


Validation of a method for localised microinjection of drugs into thalamic subregions in rats for epilepsy pharmacological studies.
To validate a method for the chronic implantation of micro-cannulae to examine the effect of drug administration to two small brain regions critical to the control of generalised seizures, the reticular nucleus of the thalamus (Rt) and the ventrobasal thalamus (VB), in a genetically epileptic rat model. Micro-cannulae guides (length 9 mm, 26G, i.d. 0.24 mm, o.d. 0.46 mm) were implanted bilaterally into either the Rt or the VB of 11- to 13-week-old Genetic Absence Epilepsy Rats from Strasbourg (GAERS) using a stereotaxic head frame. After a seven-day recovery period the animals were injected with 0.2 microl of methylene blue. The animals were allowed to move freely in their cages for a further 90 min while a post-drug EEG recording was acquired and then brains were perfused with 4% paraformaldehyde and extracted. Twenty-micrometer slices were cut on a cryostat and the site and extent of the methylene blue staining in the brain determined. The implantation co-ordinates were adjusted accordingly, and then a validation study was performed on a further cohort of rats (n=8 Rt, n=7 VB). The co-ordinates that were found to most accurately localise the Rt were: AP -3mm, ML 3.6mm, DV -5.8mm (relative to Bregma). Those that accurately localised the VB were: AP -3mm, ML 2.6mm, DV -5.5mm. In the validation study, the dye staining was measured to diffuse an average radius of 520+/-120 microm from the centre of the injection site for the 0.2 microl injection in both brain hemispheres. For the VB injections the dye remained confined within the structure, however, for the smaller Rt there was spread to surrounding structures in the axial plane. The radial diffusion for the 0.5 microl injection was similar, but more of the dye was found to spread back up the cannula tract away from the target zone. These studies have validated a method for accurate and localised injection of drugs into the VB and Rt for neuropharmacological studies in a rat model of generalised epilepsy. This method allows the measurement of localised drug effects on EEG and generalised seizure activity at these sites.


Design and synthesis of novel opioid ligands with an azabicyclo[2.2.2]octane skeleton having a 7-amide side chain and their pharmacologies.
Several derivatives with an azabicyclo[2.2.2]octane skeleton having a 7-amide side chain were synthesized. Compounds that had an electron-donating group exhibited high affinity for the μ opioid receptor while those with a bulky substituent at the 8-nitrogen atom had low affinities for all receptor types. High affinities and selectivities for the κ receptor resulted from the introduction of the longer amide side chain at the 7α-position. Our studies indicate that the orientation of the amide side chain at the 7-position within the azabicyclo[2.2.2]octane skeleton is related to selectivity for the κ receptor.


Gold nanoparticles and gold nanorods in the landscape of cancer therapy.
Cancer is a grievous disease whose treatment requires a more efficient, non-invasive therapy, associated with minimal side effects. Gold nanoparticles possessing greatly impressive optical properties have been a forerunner in bioengineered cancer therapy. This theranostic system has gained immense popularity and finds its application in the field of molecular detection, biological imaging, cancer cell targeting, etc. The photothermal property of nanoparticles, especially of gold nanorods, causes absorption of the light incident by the light source, and transforms it into heat, resulting in tumor cell destruction. This review describes the different optical features of gold nanoparticles and summarizes the advance research done for the application of gold nanoparticles and precisely gold nanorods for combating various cancers including breast, lung, colon, oral, prostate, and pancreatic cancer.


Base-pair neutral homozygotes can be discriminated by calibrated high-resolution melting of small amplicons.
Genotyping by high-resolution melting analysis of small amplicons is homogeneous and simple. However, this approach can be limited by physical and chemical components of the system that contribute to intersample melting variation. It is challenging for this method to distinguish homozygous G::C from C::G or A::T from T::A base-pair neutral variants, which comprise approximately 16% of all human single nucleotide polymorphisms (SNPs). We used internal oligonucleotide calibrators and custom analysis software to improve small amplicon (42-86 bp) genotyping on the LightScanner. Three G/C (PAH c.1155C>G, CHK2 c.1-3850G>C and candidate gene BX647987 c.261+22,290C>G) and three T/A (CPS1 c.3405-29A>T, OTC c.299-8T>A and MSH2 c.1511-9A>T) human single nucleotide variants were analyzed. Calibration improved homozygote genotyping accuracy from 91.7 to 99.7% across 1105 amplicons from 141 samples for five of the six targets. The average T(m) standard deviations of these targets decreased from 0.067 degrees C before calibration to 0.022 degrees C after calibration. We were unable to generate a small amplicon that could discriminate the BX647987 c.261+22,290C>G (rs1869458) SNP, despite reducing standard deviations from 0.086 degrees C to 0.032 degrees C. Two of the sites contained symmetric nearest neighbors adjacent to the SNPs. Unexpectedly, we were able to distinguish these homozygotes by T(m) even though current nearest neighbor models predict that the two homozygous alleles would be identical.


Association between AhR in B cells and systemic lupus erythematosus with renal damage.
Renal damage is one of the typical clinical manifestations of systemic lupus erythematosus (SLE) and few effective markers can be used to predict SLE with renal damage. Additionally, the relationship among AhR, B cells and SLE with renal damage is poorly understood. A case-control study was performed, and the clinical and laboratory data were acquired from medical records. Flow cytometry was used to analyze the expression of AhR in B cells. Both the proportion of B cells in peripheral blood lymphocytes and AhR in B cells were significantly higher in SLE patients than in healthy controls. AhR in B cells was negatively correlated with complement 3 and 4 levels but was positively correlated with SLEDAI-2 K, erythrocyte sedimentation rate, C-reactive protein and absolute lymphocyte count. Furthermore, more SLE patients suffered from renal damage and arthritis in the high-AhR B-cell group than in the low-AhR B-cell group. Interestingly, AhR in B cells was significantly increased in SLE with renal damage compared with SLE without renal damage, but no significant difference was observed between SLE with arthritis and SLE without arthritis. In addition, the area under the curve for AhR in B cells was 0.815, and its optimal cut-off level was 3.05 %, which provided a 83.3 % sensitivity and a 70.0 % specificity in predicting SLE with renal damage. AhR in B cells was correlated with SLE disease activity, and it may be a potential marker for predicting SLE with renal damage.


Assessment of Renal Risk Score and Histopathological Classification for Prediction of End-Stage Kidney Disease and Factors Associated With Change in eGFR After ANCA-Glomerulonephritis Diagnosis.
The "Renal Risk Score" (RRS) and the histopathological classification have been proposed to predict the risk of end-stage kidney disease (ESKD) in ANCA-associated glomerulonephritis (ANCA-GN). Besides, factors associated with kidney function recovery after ANCA-GN onset remain to be more extensively studied. In the present study, we analyzed the value of the RRS and of the histopathological classification for ESKD prediction. Next, we analyzed factors associated with eGFR change within the first 2 years following ANCA-GN diagnosis. We included patients from the Maine-Anjou ANCA-associated vasculitis registry with at least 6 months of follow-up. The values of ANCA-GN, histopathological classification, and RRS, and the factors associated with eGFR variations between ANCA-GN diagnosis and 2 years of follow-up were assessed. The predictive values of the histopathological classification and RRS were analyzed in 123 patients. After a median follow-up of 42 months, 33.3% patients developed ESKD. The predictive value of RRS for ESKD was greater than that of the histopathological classification. Determinants of eGFR variation were assessed in 80/123 patients with complete eGFR measurement. The median eGFR increased from ANCA-GN diagnosis to month 6 and stabilized thereafter. The only factor associated with eGFR variation in our study was eGFR at ANCA-GN diagnosis, with higher eGFR at diagnosis being associated with eGFR loss (p<0.001). The RRS has a better predictive value for ESKD than the histopathological classification. The main determinant of eGFR variation at 2 years was eGFR at ANCA-GN diagnosis. Thus, this study suggests that eGFR recovery is poorly predicted by histological damage at ANCA-GN diagnosis.


Stimulation of host DNA synthesis and induction of early antigens by ultraviolet light irradiated human cytomegalovirus.
The ultraviolet (UV-)sensitivity of the human cytomegalovirus (HCMV) genes coding for very early complement fixing and early antigens in human embryonic fibroblasts (HEF) and mouse embryonic fibroblasts (MEF) and the relation of these genes to the ability of the virus to stimulate host cell DNA synthesis were investigated. After 14 minutes of UV-irradiation of the virus inoculum only the very early complement fixing nuclear antigen (CMNA) developed in the HEF cells and only the early cytoplasmic antigen(s) was present in the MEF. In both HEF and MEF, host cell DNA synthesis was stimulated. We conclude that the ability of HCMV to stimulate host DNA synthesis is an early function of the viral genome and shows a high resistance to UV-irradiation. There is no direct correlation, however, between the ability of the virus to stimulate host cell DNA synthesis and the genes which code for the CMNA or for early cytoplasmic antigens.


Application of a chromatic confocal measurement system as new approach for in-line wet film thickness determination in continuous oral film manufacturing processes.
The key parameter of the oral film production process is the wet film thickness since it regulates the active pharmaceutical ingredient (API) content of the finished product. There is no general recommendation on how to adjust the gap height of the coating knife during the film manufacturing process to obtain the target content. Therefore, trial and error approaches are common to determine the surplus of drug for every newly developed formulation. This wastes resources, money and time and calls for an adequate in-line tool for wet film thickness measurement during the film manufacturing process to ensure consistent quality. In this work, a chromatic confocal optical probe was implemented into a continuous oral film manufacturing process on a pilot-scale coating bench. The optical probe allows a non-destructive and contactless wet film thickness measurement. The validation of the method showed good results. Linearity was demonstrated over a wide range of film thicknesses (R2 = 0.999). A good precision between different films was revealed by a coefficient of variation smaller than 2%. The robustness investigations showed that the method is applicable for transparent and non-transparent film forming masses. Furthermore, coloring agents, particles in the polymer mass and different viscosities do not influence the thickness measurement.


Luteinization to oocyte retrieval delay in women in whom multiple follicular growth was induced as part of an in vitro fertilization/gamete intrafallopian transfer program.
Different luteinization-to-oocyte-retrieval delays were examined in 64 IVF/GIFT patients. Multiple follicular stimulation was induced with combined therapy of hMG and GnRH-a to create continuous pituitary suppression and thus eliminate endogenous gonadotropin fluctuations. The results demonstrated that with this induction regime, the time interval can be extended to 39 hours without preoperative ovulation and with improved oocyte maturity.


Aryl-triazole foldamers incorporating a pyridinium motif for halide anion binding in aqueous media.
Aryl-triazole oligomers incorporating a pyridinium motif have been synthesized from their pyridine precursors. Anion binding studies show that methylation of the pyridine units can significantly enhance the halide anion affinities of the folded oligomers so that the foldamers are capable of binding halide anions in aqueous solutions.


Recurrent oral carcinoma in Ferguson-Smith disease.
A patient with multiple self-healing squamous cell carcinomas of the skin is presented. Mucous membrane involvement in this condition is very uncommon and the lesions may show aggressive behaviour.


Ubiquitin, a central component of selective cytoplasmic proteolysis, is linked to proteins residing at the locus of non-selective proteolysis, the vacuole.
Ubiquitin, an evolutionary highly conserved protein, is known to be involved in selective proteolysis in the cytoplasm. Here we show that ubiquitin-protein conjugates are also found in the yeast vacuole. Mutants defective in the major vacuolar endopeptidases, proteinase yscA and yscB, lead to accumulation of ubiquitin-protein conjugates in this cellular organelle.


Proliferation and dissemination of killer meiotic drive loci.
Killer meiotic drive elements are selfish genetic entities that manipulate the sexual cycle to promote their own inheritance via destructive means. Two broad classes are sperm killers, typical of animals and plants, and spore killers, which are present in ascomycete fungi. Killer meiotic drive systems operate via toxins that destroy or disable meiotic products bearing the alternative allele. To avoid suicidal autotargeting, cells that bear these selfish elements must either lack the toxin target, or express an antidote. Historically, these systems were presumed to require large nonrecombining haplotypes to link multiple functional interacting loci. However, recent advances on fungal spore killers reveal that numerous systems are enacted by single genes, and similar molecular genetic studies in Drosophila pinpoint individual loci that distort gamete sex. Notably, many meiotic drivers duplicate readily, forming gene families that can have complex interactions within and between species, and providing substrates for their rapid functional diversification. Here, we summarize the known families of meiotic drivers in fungi and fruit flies, and highlight shared principles about their evolution and proliferation that promote the spread of these noxious genes.


The significance of physical development on the blood pressure curve of children between 6 and 9 years of age and its relationship with familial aggregation.
Arterial blood pressure values of 3109 children between 6 and 9 years of age in the city of Rio de Janeiro were analysed according to low, medium and high socio-economic levels. At medium and high socio-economic levels, arterial pressures, weight and height were higher than those observed in children of a lower socio-economic level. Blood pressure was best correlated to body weight. The aggregation of arterial pressure, weight and height was compared between family members of children with percentiles below 50 and above 90. Systolic blood pressure and weight were significantly lower in family members with a percentile below 50. We conclude that there is a need for a long-term follow-up of these children and their families, in order to improve our knowledge of the mechanisms involved in the origin of arterial hypertension.


The dopamine autoreceptor agonist B-HT 920 markedly stimulates sexual behavior in male rats.
B-HT 920, a selective agonist at dopamine (DA) autoreceptors, strongly increased the incidence of penile erections (PE) in male rats, an effect which was dose-related and antagonized by haloperidol. B-HT 920 at 100 and 200 micrograms/kg i.p. significantly altered the copulatory pattern of sexually active male rats, reducing the number of mounts and intromissions as well as the latency to the first ejaculation, a stimulant effect which was confirmed in sluggish males at a dose of 100 micrograms/kg.


Social anxiety disorders in clinical practice: differentiating social phobia from avoidant personality disorder.
To outline the problems around overlap between social phobia (SAD) and avoidant personality disorder (AVPD) and provide guidelines that may assist clinicians to differentiate these conditions. A constellation of symptoms can be identified that may distinguish AVPD from SAD, with key features being a strong and pervasively negative self-concept, a view of rejection as equating to a global evaluation of the individual as being of little worth and a sense of not fitting in socially that dates from early childhood. It is important to identify the presence of AVPD in order to anticipate potential problems with engagement and retention in therapy, to target treatment interventions and optimise outcome.


Effect of dilution rate and nutrients addition on the fermentative capability and synthesis of aromatic compounds of two indigenous strains of Saccharomyces cerevisiae in continuous cultures fed with Agave tequilana juice.
Knowledge of physiological behavior of indigenous tequila yeast used in fermentation process is still limited. Yeasts have significant impact on the productivity fermentation process as well as the sensorial characteristics of the alcoholic beverage. For these reasons a better knowledge of the physiological and metabolic features of these yeasts is required. The effects of dilution rate, nitrogen and phosphorus source addition and micro-aeration on growth, fermentation and synthesis of volatile compounds of two native Saccharomyces cerevisiae strains, cultured in continuous fed with Agave tequilana juice were studied. For S1 and S2 strains, maximal concentrations of biomass, ethanol, consumed sugars, alcohols and esters were obtained at 0.04 h⁻¹. Those concentrations quickly decreased as D increased. For S. cerevisiae S1 cultures (at D=0.08 h⁻¹) supplemented with ammonium phosphate (AP) from 1 to 4 g/L, concentrations of residual sugars decreased from 29.42 to 17.60 g/L and ethanol increased from 29.63 to 40.08 g/L, respectively. The S1 culture supplemented with AP was then micro-aerated from 0 to 0.02 vvm, improving all the kinetics parameters: biomass, ethanol and glycerol concentrations increased from 5.66, 40.08 and 3.11 g/L to 8.04, 45.91 and 4.88 g/L; residual sugars decreased from 17.67 g/L to 4.48 g/L; and rates of productions of biomass and ethanol, and consumption of sugars increased from 0.45, 3.21 and 7.33 g/L·h to 0.64, 3.67 and 8.38 g/L·h, respectively. Concentrations of volatile compounds were also influenced by the micro-aeration rate. Ester and alcohol concentrations were higher, in none aerated and in aerated cultures respectively.


Synthesis and biological activity of Ub2 derived peptides as potential host-directed antitubercular therapy.
The correlation of mycobactericidal property of macrophages with its potential to deliver bacteria to hydrolytic lysosomes, augmented with ubiquitin-derived peptides (Ub2), activates the process of autophagy. This leads to the formation of phagolysosomes supported by factor involving increased cationic charges which regulate the acidic pH causing elimination of Mycobacterium. To better understand this interaction of cationic-rich ubiquitin-derived peptides with mycobacteria and to identify putative mycobacterial intrinsic resistance mechanisms for phagolysosome formation, we have synthesized a new series of Ub2 peptides, wherein the Gly residues are replaced with azaGly with the aim to improve metabolic stability. In addition to that a new methodology is reported for the synthesis of heteroaryl tethered peptides using azaGly as a linker. We have demonstrated that positive puncta (directly proportional to the acidification of lysosome) in cytosol was significantly increased after 6 hours on the treatment of macrophage with Ub2 peptide derivatives (1, 6, 10, and 11) causing the higher intensity of lysosome observed through LysoTracker Red Dye. The circular dichroism spectral studies are carried out in water and water:TFE mixture and demonstrated that the Ub2 peptides have helix-forming tendency in the presence of TFE. The recognizable intracellular killing of Mycobacterium tuberculosis by Ub2 peptides provides a new approach for host-directed therapy.


A binocular perception deficit characterizes prey pursuit in developing mice.
Integration of binocular information at the cellular level has long been studied in the mouse model to uncover the fundamental developmental mechanisms underlying mammalian vision. However, we lack an understanding of the corresponding ontogeny of visual behavior in mice that relies on binocular integration. To address this major outstanding question, we quantified the natural visually guided behavior of postnatal day 21 (P21) and adult mice using a live prey capture assay and a computerized-spontaneous perception of objects task (C-SPOT). We found a robust and specific binocular visual field processing deficit in P21 mice as compared to adults that corresponded to a selective increase in c-Fos expression in the anterior superior colliculus (SC) of the juveniles after C-SPOT. These data link a specific binocular perception deficit in developing mice to activity changes in the SC.


Development of a simple, accurate SPME-based method for assay of VOCs in column breakthrough experiments.
Solid-phase microextraction (SPME) with gas chromatography is to be used for assay of effluent liquid samples from soil column experiments associated with VOC fate/transport studies. One goal of the fate/transport studies is to develop accurate, highly reproducible column breakthrough curves for 1,2-cis-dichloroethylene (cis-DCE) and trichloroethylene (TCE) to better understand interactions with selected natural solid phases. For SPME, the influences of the sample equilibration time, extraction temperature and the ratio of volume of sample bottle to that of the liquid sample (V(T)/V(w)) are the critical factors that could influence accuracy and precision of the measured results. Equilibrium between the gas phase and liquid phase was attained after 200 min of equilibration time. The temperature must be carefully controlled due to variation of both the Henry's constant (K(h)) and the fibre/gas phase distribution coefficient (K(fg)). K(h) decreases with decreasing temperature while K(fg) increases. Low V(T)/V(w) yields better sensitivity but results in analyte losses and negative bias of the resultant assay. High V(T)/V(w) ratio yields reduced sensitivity but analyte losses were found to be minimal, leading to better accuracy and reproducibility. A fast SPME method was achieved, 5 min for SPME extraction and 3.10 min for GC analysis. A linear calibration function in the gas phase was developed to analyse the breakthrough curve data, linear between a range of 0.9-236 microgl(-1), and a detection limit lower than 5 microgl(-1).


Patches and the responses of lake benthos to sunfish nest-building.
Males of three sunfish species (Centrarchidae) construct nests for spawning and often share them sequentially in the littoral zone of a 4-hectare lake in New York State. To determine spatial and temporal effects of this reproductive behavior on zoobenthos in "patches," I sampled bottom assemblages from inside nests and from adjacent (<1 m from nest perimeter), relatively undisturbed sites during the reproductive season of 1986 and immediately prior to nest-building in 1987. The reproductive behaviors of largemouth bass (Micropterus salmoides), redbreast sunfish (Lepomis auritus), and pumpkinseeds (L. gibbosus) altered relative abundances and significantly decreased benthic invertebrate diversity and density. These effects were extremely pronounced during the reproductive season and were partially detectable the following year. Community changes were probably the result of both bioturbation (modification of sediment size and organic content) and predation. The ecosystem-wide effects of nest-building are evaluated in terms of the number, distribution, and longevity of patches.


Amyotrophic lateral sclerosis in Nova Scotia.
A study of ALS incidence in Nova Scotia for a ten year period was carried out with the cooperation of neurologists, neurosurgeons, physiatrists and major medical hospitals. Over a ten year period 161 cases occurred, an incidence of 1.95 per 100,000 population in a province with a population of 825,000. There was an increase in incidence over the ten year period, with an incidence for the first five years of study of 1.50 per 100,000, and 2.40 per 100,000 for the latter five years. The peak incidence in 1981-1982 was 2.66 per 100,000. Although this is the highest incidence in community studies of ALS (excepting the unusual circumstances in Guam), it may reflect the realistic incidence of ALS rather than an unusual focus, as ascertainment was likely greater than in many other studies. The more significant finding is the suggestion of an increasing incidence, as noted in other studies in the U.S., England and Israel.


Fast magnetization transfer and apparent T1 imaging using a short saturation pulse with and without inversion preparation.
Measurements of magnetization transfer (MT) metrics, such as the steady-state (SS) MT ratio and apparent longitudinal relaxation rate require multiple MT irradiation durations and a long experimental time. To overcome these problems, we propose a novel method using a short MT off-resonance pulse with and without on-resonance inversion preparation pulse. Computer simulations were performed to examine the accuracy of the proposed method and to find the optimal off-resonance irradiation pulse duration (Tirad) and power level (ω1). Our approach, with echo planar imaging data acquisition, was applied to animals at 9.4 T and humans at 3 T with ω1/2π = 100 Hz and 177 Hz, respectively. Steady-state MT ratio and relaxation rate were obtained from a pair of MT images at a Tirad, with and without inversion. For Tirad ≥ 0.4 s, steady-state MT ratio, and relaxation rate measured at any single Tirad agreed well with those of the conventional fitting method that uses multiple Tirad. Our simulation indicates that a higher ω1 can use a shorter Tirad. Steady-state MT ratio and relaxation rate can be determined from MT data with only one, short Tirad by incorporation of an inversion prepulse. This MT imaging approach is simple, fast, and easily implementable.


Massive hemothorax due to bleeding from thoracic spinal fractures: a case series and systematic review.
Massive hemothorax secondary to thoracic spinal fractures is rare, and its clinical characteristics, treatment, and prognosis are unknown. We present two cases of thoracic spinal fracture-induced massive hemothorax and a systematic review of previously reported cases. This study included patients with traumatic hemothorax from thoracic spinal fractures at a Japanese tertiary care hospital. A systematic review of published cases was undertaken through searches in PubMed, EMBASE, and ICHUSHI from inception to October 13, 2019. Case 1: An 81-year-old man developed hemodynamic instability from a right hemothorax with multiple rib fractures following a pedestrian-vehicle accident; > 1500 mL blood was evacuated through the intercostal drain. Thoracotomy showed hemorrhage from a T8-burst fracture, and gauze packing was used for hemostasis. Case 2: A 64-year-old man with right hemothorax and hypotension after a fall from height had hemorrhage from a T7-burst fracture, detected on thoracotomy, which was sealed with bone wax. Hypotension recurred during transfer; re-thoracotomy showed bleeding from a T7 fracture, which was packed with bone wax and gauze for hemostasis. The systematic review identified 10 similar cases and analyzed 12 cases, including the abovementioned cases. Inferior part of thoracic spines was prone to injury and induced right-sided hemothorax. Most patients developed hemodynamic instability, and some sustained intra-transfer hemorrhage; direct compression (gauze packing, bone wax, and hemostatic agents) was the commonest hemostatic procedure. The mortality rate was 33.3%. Hemothorax due to thoracic spinal fracture can be fatal. Thoracotomy with direct compression is necessary in hemodynamically unstable patients.


Successful palliation of acute superior vena caval obstruction after the Senning operation.
Superior vena caval obstruction after an intraatrial baffle procedure can lead to acute cerebral complications and hence requires immediate management. We present a case of successful palliation of acute superior vena caval obstruction after a Senning procedure by establishing a venous shunt between the innominate vein and pulmonary artery. This technique resulted in immediate hemodynamic and functional improvement that subsequently allowed for the enlargement of the superior vena cava-right atrial junction with a pericardial patch.


Massive hemoptysis.
Massive hemoptysis is an uncommon but life-threatening emergency. The loss of at least 600 mL of blood within a 48-hour period has been associated with a high mortality rate. Initial stabilization including airway and ventilation management, IV fluids, oxygen, and laboratory and radiographic studies should be done in the ED. Bronchoscopy and angiography are initial diagnostic manuevers that also may be therapeutic. Surgical therapy is reserved for patients with adequate pulmonary reserve and localized sources of bleeding.


Evaluation of the energetic contribution of an ionic network to beta-sheet stability.
We have evaluated the interaction energy of a three-residue ionic network constructed on the beta-sheet surface of protein G using double mutant cycles. Although the two individual ion pairs were each stabilizing by 0.6 kcal/mol, the excess gain in stability for the triad was small (0.06 kcal/mol).


Serum osteocalcin radioimmunoassay in bone diseases.
Osteocalcin (or bone Gla protein, BGP) is a non-collagenous vitamin K-dependent protein accounting for 1-2% of the total bone proteins. It represents a specific index of osteoblastic activity and directly reflects the bone turnover. Serum levels of osteocalcin were measured by a radioimmunoassay method. In 40 postmenopausal women with osteoporosis, mean serum BGP levels were lower than the normal range (3.69 +/- 1.35 ng/ml), whereas they significantly increased in 7 patients with osteomalacia (10.48 +/- 3.05 ng/ml), in 12 patients with secondary hyperparathyroidism (11.1 +/- 4.9 ng/ml) and in 41 patients with Paget's disease (12.09 +/- 6.5 ng/ml). Four patients with primary hyperparathyroidism showed very high BGP levels (64.0 +/- 32.3 ng/ml), which strikingly fell after the surgical removal of a parathyroid adenoma. These results confirm that the quantitation of serum osteocalcin is a specific and sensitive method in the diagnosis of bone disease, represents a useful index of bone turnover and is particularly helpful in the follow-up of patients with treated bone disease.


Utility of Porous Glass with a new long-chain alkylamine spacer arm as a solid support for synthesis of oligodeoxyribonucleotides via the phosphoramidite method.
Porous Glass with an [[N-(2-aminoethyl)aminomethyl]phenyl]ethylsilyl spacer arm serving as a useful solid support in the synthesis of oligodeoxyribonucleotides via the phosphoramidite strategy.


Cerebral protection in left atrial appendage closure in the presence of appendage thrombosis.
Presence of thrombus in the left atrial appendage (LAA) remains a severe contraindication to the percutaneous left atrial appendage closure procedure (LAAC), due to increased embolic risk. Recently, the experience developed in cerebral protection device in transcatheter aortic valve implantation (TAVI) procedure was translated in LAAC to address this issue. To evaluate efficacy and safety of Sentinel cerebral protection system (CPS) in supporting LAAC in real-world patient with persistent LAA thrombus. The study retrospectively enrolled consecutive patients with non-valvular atrial fibrillation (NVAF) and thrombus in LAA who underwent LAAC supported by Sentinel CPS in seven European high-volume centres. Twenty-seven patients were included with a median age of 69.1 ± 9.7 years old, with median CHA2 DS2 -VASc and HAS-BLEED scores 3 [2-5] and 3 [2.75-4], respectively. Technical and procedural success was achieved in all patients. No periprocedural TIA, stroke, or supra-aortic trunks dissection was recorded. In this multicenter registry, LAAC supported by Sentinel CPS in patients with LAA persistent thrombus seems to be a safe and efficacious treatment.


Challenges and Opportunities with Predicting in Vivo Phase II Metabolism via Glucuronidation from in Vitro Data.
Glucuronidation is the most important phase II metabolic pathway which is responsible for the clearance of many endogenous and exogenous compounds. To better understand the elimination process for compounds undergoing glucuronidation and identify compounds with desirable in vivo pharmacokinetic properties, many efforts have been made to predict in vivo glucuronidation using in vitro data. In this article, we reviewed typical approaches used in previous predictions. The problems and challenges in prediction of glucuronidation were discussed. Besides that different incubation conditions can affect the prediction accuracy, other factors including efflux / uptake transporters, enterohepatic recycling, and deglucuronidation reactions also contribute to the disposition of glucuronides and make the prediction more difficult. PBPK modeling, which can describe more complicated process in vivo, is a promising prediction strategy which may greatly improve the prediction of glucuronidation and potential DDIs involving glucuronidation. Based on previous studies, we proposed a transport-glucuronidation classification system, which was built based on the kinetics of both glucuronidation and transport of the glucuronide. This system could be a very useful tool to achieve better in vivo predictions.


The effect of absorbable gelatin sponge on experimental microvascular anastomoses.
Following a pilot study a controlled trial was carried out to assess the effect of absorbable gelatin sponge on reducing the bleeding from microvascular anastomoses of the femoral arteries and veins of rats. A model was used which approximated to the clinical situation and the bleeding time was significantly reduced in arteries and veins. There was no significant difference in patency rates or wound infection.


Depth of focus and functional results in patients with multifocal intraocular lenses: a long-term follow-up.
We did a follow-up study of 35 patients who had phacoemulsification with implantation of diffractive, multifocal intraocular lenses (IOLs) 2.5 years ago. The patients had an average age of 58.2 +/- 9.6 years. Fifty multifocal IOLs were implanted (3M 815LE, n = 45; 3M 825X, n = 5). Refractive values, visual acuity, depth of focus, and the dependence on corrective glasses were evaluated. Eighty percent of the spherical equivalents ranged from -1 to +1 D (mean 0.25 +/- 0.71). In 76% the cylindric power was not higher than 1 D (mean 0.87 +/- 0.64). With distance correction, 100% of the patients achieved a visual acuity of 20/40 or better. Without correction, 86% achieved 20/40 or better. Near acuity of 20/40 or better with only distance correction was achieved in 98% of patients. Ninety-four percent had a near acuity of 20/40 or better without any correction. The depth of focus was significantly better in the multifocal group than in a control group of patients with monofocal IOLs (P < .01). Although 90% of the patients had good uncorrected visual acuity results of 20/40 or better for near and distance, 58% preferred glasses for distance and/or near.


Adrenal haemangioblastoma presenting as phaeochromocytoma: a rare manifestation of extraneural hemangioblastoma.
Adrenal haemangioblastoma presenting clinically as pheochromocytoma is a rare manifestation of extraneural haemangioblastoma. We present an unusual case of von Hippel-Lindau (VHL) disease that had adrenal and cerebellar haemangioblastoma with multiple renal cysts, and a review of the literature. Unlike the usual manifestations of secondary polycythemia or increased intracranial pressure and hydrocephalus due to cerebellar lesion, this 36-year-old male presented with hypertension. Investigations revealed right suprarenal mass with raised urinary catecholamines and serum vanillylmandelic acid (VMA) levels, apparently confirming the clinical diagnosis of phaeochromocytoma. Histopathology of the biopsy specimen showed features of haemangioblastoma, which was confirmed by immunohistochemistry using antibodies to neuron specific enolase and aquaporin-1. Based on this, the patient was screened for possible features of VHL, which revealed cerebellar haemangioblastoma and multiple renal cysts with angiomatous lesion. Postoperative follow-up showed normal levels of catecholamines without any symptoms of phaeochromocytoma. Adrenal haemangioblastoma is a rare entity with only four cases reported in the literature. Surgical removal is the treatment of choice. However, screening for other possible features of VHL, even in the absence of clinical features, is essential to exclude other potential lesions.


Poly(d,l-lactide)/polyethylene glycol micro/nanofiber mats as paclitaxel-eluting carriers: preparation and characterization of fibers, in vitro drug release, antiangiogenic activity and tumor recurrence prevention.
Poly(d,l-lactide)/polyethylene glycol (PLA/PEG) micro/nanofibers loaded with paclitaxel (PTX, 10 wt%) were prepared by needless electrospinning technology, which allows large scale production for real medicinal practice. The fiber structure and properties were investigated by several methods including scanning electron microscopy, nitrogen adsorption/desorption isotherm measurements, differential scanning calorimetry, and X-ray diffraction measurements to examine their morphology (fiber diameter distribution, specific surface area, and total pore volume), composition, drug-loading efficiency, and physical state. An HPLC-UV method was optimized and validated to quantify in vitro PTX release into PBS. The results showed that the addition of PEG into PLA fibers promoted the release of higher amounts of hydrophobic PTX over prolonged time periods compared to fibers without PEG. An in vitro cell assay demonstrated the biocompatibility of PLA/PEG fibrous materials and showed significant cytotoxicity of PTX-loaded PLA/PEG fibers against a human fibrosarcoma HT1080 cell line. The chick chorioallantoic membrane assay proved that PTX-loaded fibers exhibited antiangiogenic activity, with a pronounced effect in the case of the PEG-containing fibers. In vivo evaluation of PTX-loaded PLA/PEG fibers in a human fibrosarcoma recurrence model showed statistically significant inhibition in tumor incidence and growth after primary tumor resection compared to other treatment groups.


Community health services and health care utilization in rural Bangladesh.
The study, which is based on data from two household level health surveys conducted in 1976 and 1987 in the Companiganj area of rural Bangladesh, examines the premise that the utilization of public health care services can be increased by increasing the availability and accessibility of effective medicines to the public and by improving the disease recognition and management by the health practitioners. The results of the study suggest that the availability and accessability of modern effective medicines through the provision of decentralized community-based rural health services, by a well-trained and well-managed field personnel structure, had an incremental impact on the utilization of modern health care from a rural health center and its subcenters. The study further reveals that, in 1976 as well as in 1987, the overwhelming majority of the rural Bangladesh population were using modern Western medical practitioners, although most of these practitioners were informally trained or self-trained without any formal medical degrees or training. It is concluded that the persons responsible for health program planning and health program implementation need to ensure that the access to basic public health care services be made broad enough to cover the majority of the rural population through a system of decentralized curative and preventive services, as well as through a system of adequate training and deployment of health professionals, including training programs to improve the quality of medical services offered by the informal and self-trained practitioners of modern medicines.


Identification of two lipid phosphatases that regulate sphingosine-1-phosphate cellular uptake and recycling.
Sphingosine-1-phosphate (S1P) is a sphingolipid metabolite that serves as a potent extracellular signaling molecule. Metabolic regulation of extracellular S1P levels impacts key cellular activities through altered S1P receptor signaling. Although the pathway through which S1P is degraded within the cell and thereby eliminated from reuse has been previously described, the mechanism used for S1P cellular uptake and the subsequent recycling of its sphingoid base into the sphingolipid synthesis pathway is not completely understood. To identify the genes within this S1P uptake and recycling pathway, we performed a genome-wide CRISPR/Cas9 KO screen using a positive-selection scheme with Shiga toxin, which binds a cell-surface glycosphingolipid receptor, globotriaosylceramide (Gb3), and causes lethality upon internalization. The screen was performed in HeLa cells with their sphingolipid de novo pathway disabled so that Gb3 cell-surface expression was dependent on salvage of the sphingoid base of S1P taken up from the medium. The screen identified a suite of genes necessary for S1P uptake and the recycling of its sphingoid base to synthesize Gb3, including two lipid phosphatases, PLPP3 (phospholipid phosphatase 3) and SGPP1 (S1P phosphatase 1). The results delineate a pathway in which plasma membrane-bound PLPP3 dephosphorylates extracellular S1P to sphingosine, which then enters cells and is rephosphorylated to S1P by the sphingosine kinases. This rephosphorylation step is important to regenerate intracellular S1P as a branch-point substrate that can be routed either for dephosphorylation to salvage sphingosine for recycling into complex sphingolipid synthesis or for degradation to remove it from the sphingolipid synthesis pathway.


Voyager 1 in the foreshock, termination shock, and heliosheath.
Voyager 1 (V1) began measuring precursor energetic ions and electrons from the heliospheric termination shock (TS) in July 2002. During the ensuing 2.5 years, average particle intensities rose as V1 penetrated deeper into the energetic particle foreshock of the TS. Throughout 2004, V1 observed even larger, fluctuating intensities of ions from 40 kiloelectron volts (keV) to >/=50 megaelectron volts per nucleon and of electrons from >26 keV to >/=350 keV. On day 350 of 2004 (2004/350), V1 observed an intensity spike of ions and electrons that was followed by a sustained factor of 10 increase at the lowest energies and lesser increases at higher energies, larger than any intensities since V1 was at 15 astronomical units in 1982. The estimated solar wind radial flow speed was positive (outward) at approximately +100 kilometers per second (km s(-1)) from 2004/352 until 2005/018, when the radial flows became predominantly negative (sunward) and fluctuated between approximately -50 and 0 km s(-1) until about 2005/110; they then became more positive, with recent values (2005/179) of approximately +50 km s(-1). The energetic proton spectrum averaged over the postshock period is apparently dominated by strongly heated interstellar pickup ions. We interpret these observations as evidence that V1 was crossed by the TS on 2004/351 (during a tracking gap) at 94.0 astronomical units, evidently as the shock was moving radially inward in response to decreasing solar wind ram pressure, and that V1 has remained in the heliosheath until at least mid-2005.


Cloning and expression of cDNA encoding Xenopus laevis bone morphogenetic protein-1 during early embryonic development.
The Xenopus laevis DNA fragment encoding a protein homologous with human bone morphogenetic protein-1 (BMP-1) was amplified by polymerase chain reaction (PCR) from cDNA derived from stage 26 (st.26) embryonic RNA. Subsequently this fragment was used as a probe to isolate cDNA clones by screening of a X. laevis st.24 embryonic cDNA library. Two partial clones (22 and 63) were obtained and the missing 5'-end of the clone 22 was extended by the anchored PCR technique. The nucleotide sequence of the resulting clone (22AN) contained an open reading frame coding for a protein with 707 deduced amino acids. Three sizes of mRNA (2.9, 5.2 and 6.6 kb) were detected in blastula (st.9) and early gastrula (st.10) embryos, and in hatched tadpole (st.40), but little or no expression was observed in morula (st.7) and late gastrula (st.12) embryos, suggesting a physiological role(s) of X.laevis BMP-1 in normal embryonic development.


Phenotypical and molecular characterization of Microsporum canis strains in north-east Brazil.
This study investigated the possible correlation between the phenotypical and genotypical characteristics of Microsporum canis isolated from cats and dogs in north-east Brazil. The mycological study was conducted by direct microscopic examination and by fungal culture. Polymerase chain reaction-restriction enzyme analysis and random amplification of polymorphic DNA techniques were used for the genotypical analysis. The morphological analysis showed a considerable diversity of colonies as well as different morphologies of conidia, despite the M. canis strains having been isolated under the same conditions. However, the molecular analysis showed that all analysed strains are genetically similar. This study, based on phenotypical and molecular analysis, evidences the wide spectrum of phenotypical variations in M. canis in contrast to the stable genotypes of such dermatophytes. The findings of this study indicate that M. canis isolated from cats and dogs with dermatophytosis in north-east Brazil may be clones, well adapted to the conditions of this region, despite M. canis showing different morphological features.


Maternal mood and anxiety effects on the fetal nonstress test.
Previous studies have suggested that acute mood states may influence levels of fetal activity. Because the fetal nonstress test relies on markers of fetal activity to suggest fetal wellbeing, its interpretation may be affected by maternal mood. This study sought to determine if there are differences in nonstress test characteristics between pregnant individuals with and without symptoms of mood disorder. In this prospective cohort study, we recruited pregnant individuals undergoing nonstress test in the third trimester and compared the results of the nonstress test between pregnant individuals with scores above and below the cutoff values on validated screening questionnaires for depression and anxiety symptoms, the Patient Health Questionnaire-9 (PHQ-9) and Generalized Anxiety Disorder 7-item scale (GAD-7). Demographic information was collected from each participant at the time of recruitment, and medical information was extracted from the electronic medical record. A total of 68 pregnant individuals were enrolled, 10 (15%) of which screened positive for perinatal mood disorders. There was no significant difference in means of time to reactivity (15.6 [4.8] minutes vs 15.0 [8.0] minutes, P=.77), number of accelerations (0.16/min [0.08] vs 0.16/min [0.10], P>.95), number of fetal movements (17.0 [14.7] vs 19.7 [20.4], P=.62), heart rate baseline (138.0 [7.5] bpm vs 139.2 [9.0] bpm, P=.67) or variability (8.5 [2.5] bpm vs 9.1 (4.3) bpm, P=.51) between pregnant individuals who screen positive for mood disorders and those who did not. Fetal heart rate patterns are similar in pregnant individuals with and without symptoms of mood disorder. The results provide reassurance that acute symptoms of anxiety and depression do not have significant effects on the fetal nonstress test.


Effect of lead, calcium, iron, zinc, copper and magnesium on anemia in children with BLLs ≥ 100 μg/L.
Adverse effects of lead exposure on children's health have been demonstrated. While studies have examined the relationship between iron status and low-level lead exposure in children with blood lead levels (BLLs) < 100 μg/L, few have investigated the association between blood lead and other trace elements and anemia in children with BLLs ≥ 100 μg/L. This study aimed to assess the levels of lead, iron, copper, zinc, magnesium, and calcium in children aged 0-14 with BLLs≥ 100 μg/L between 2009 and 2021, and to examine the relationship between blood lead, trace elements and anemia. A total of 11,541 children with BLLs ≥ 100 μg/L were included in this study. Venous blood samples were collected to measure blood lead levels, hemoglobin levels, and trace element levels. According to the World Health Organization standard, outpatients with hemoglobin levels < 110 g / L were defined as having anemia. The study results found that high BLLs and blood calcium had a negative influence on Hb with odds ratios (95% confidence interval) of 1.411(1.208, 1.649) and 1.219(1.043, 1.424). High blood iron had a positive influence on Hb with odds ratios of 0.421(0.355, 0.499). The results suggest that the risk of anemia rose significantly with higher BLLs, blood copper, and blood calcium levels, and decreases considerably with higher blood iron levels.


Voltammetric investigation of cytochrome c on gold coated with a self-assembled glutathione monolayer.
The direct, reversible electrochemistry of horse-heart cytochrome c (cyt. c) was realized on a self-assembled glutathione (GSH) monolayer modified Au electrode. The voltammetric responses of cyt. c on GSH/Au electrode were found to be affected by pH during the electrode modification, metal ions and surfactants. Using potassium ferricyanide [K4Fe(CN)6] as a probe, these effects on the voltammetric responses of cyt. c were characterized by electrochemical methods. It was found that the pH during the electrode modification, metallic ions and surfactants changed GSH monolayer's charge state and the conformation on the electrode surface, and resulted in the influence on the voltammetric responses of cyt. c. The experimental results provided us information to understand the mechanism of the interfacial electron transfer of electrode-protein, as well as the electron transfer of cyt. c in life system.


An integrated assessment of the Good Environmental Status of Mediterranean Marine Protected Areas.
Local, regional and global targets have been set to halt marine biodiversity loss. Europe has set its own policy targets to achieve Good Environmental Status (GES) of marine ecosystems by implementing the Marine Strategy Framework Directive (MSFD) across member states. We combined an extensive dataset across five Mediterranean ecoregions including 26 Marine Protected Areas (MPAs), their reference unprotected areas, and a no-trawl case study. Our aim was to assess if MPAs reach GES, if their effects are local or can be detected at ecoregion level or up to a Mediterranean scale, and which are the ecosystem components driving GES achievement. This was undertaken by using the analytical tool NEAT (Nested Environmental status Assessment Tool), which allows an integrated assessment of the status of marine systems. We adopted an ecosystem approach by integrating data from several ecosystem components: the seagrass Posidonia oceanica, macroalgae, sea urchins and fish. Thresholds to define the GES were set by dedicated workshops and literature review. In the Western Mediterranean, most MPAs are in good/high status, with P. oceanica and fish driving this result within MPAs. However, GES is achieved only at a local level, and the Mediterranean Sea, as a whole, results in a moderate environmental status. Macroalgal forests are overall in bad condition, confirming their status at risk. The results are significantly affected by the assumption that discrete observations over small spatial scales are representative of the total extension investigated. This calls for large-scale, dedicated assessments to realistically detect environmental status changes under different conditions. Understanding MPAs effectiveness in reaching GES is crucial to assess their role as sentinel observatories of marine systems. MPAs and trawling bans can locally contribute to the attainment of GES and to the fulfillment of the MSFD objectives. Building confidence in setting thresholds between GES and non-GES, investing in long-term monitoring, increasing the spatial extent of sampling areas, rethinking and broadening the scope of complementary tools of protection (e.g., Natura 2000 Sites), are indicated as solutions to ameliorate the status of the basin.


Possible role of DT-diaphorase in the bioactivation of antitumor quinones.
Menadione derivatives that are toxic to tumor cells are believed to be reduced intracellularly to species that react with DNA. In this communication, we report evidence that one of these derivatives, 3-bromomethylmenadione, is reduced by DT-diaphorases present in rat liver cytosol and in rat 9L brain tumor cells. Dicoumarol, an inhibitor of DT-diaphorases was found to inhibit both the reduction of 3-bromomethylmenadione and its mutagenicity to Salmonella typhimurium TA 97. Homogenates of rat 9L cells were found to contain relatively high levels of DT-diaphorase, suggesting that these tumor cells may be relatively sensitive to antitumor quinones that are activated by this enzyme.


RGS12 is required for the maintenance of mitochondrial function during skeletal development.
Mitochondrial morphology and function are crucial for tissue homeostasis, such as for skeletal development, but the cellular and molecular mechanisms remain unclear. Here, we provide evidence that regulator of G-protein signaling 12 (RGS12) is present in the mitochondria of primary chondrocytes and cartilage tissues. Deletion of RGS12 in type II collagen-positive cells led to a significant decrease in mitochondrial number, membrane potential, and oxidative phosphorylation function. Mechanistically, RGS12 promoted the function of ATP5A as an enhancer of tyrosine phosphorylation. Mice with RGS12 deficiency in the chondrocyte lineage showed serious body retardation, decreased bone mass, and chondrocyte apoptosis due to the defective activity of ATP synthase. To our knowledge, this is the first report that RGS12 is required for maintaining the function of mitochondria, which may allow it to orchestrate responses to cellular homeostasis.


Vaxi-DL: A web-based deep learning server to identify potential vaccine candidates.
The development of a new vaccine is a challenging exercise involving several steps including computational studies, experimental work, and animal studies followed by clinical studies. To accelerate the process, in silico screening is frequently used for antigen identification. Here, we present Vaxi-DL, web-based deep learning (DL) software that evaluates the potential of protein sequences to serve as vaccine target antigens. Four different DL pathogen models were trained to predict target antigens in bacteria, protozoa, fungi, and viruses that cause infectious diseases in humans. Datasets containing antigenic and non-antigenic sequences were derived from known vaccine candidates and the Protegen database. Biological and physicochemical properties were computed for the datasets using publicly available bioinformatics tools. For each of the four pathogen models, the datasets were divided into training, validation, and testing subsets and then scaled and normalised. The models were constructed using Fully Connected Layers (FCLs), hyper-tuned, and trained using the training subset. Accuracy, sensitivity, specificity, precision, recall, and AUC (Area under the Curve) were used as metrics to assess the performance of these models. The models were benchmarked using independent datasets of known target antigens against other prediction tools such as VaxiJen and Vaxign-ML. We also tested Vaxi-DL on 219 known potential vaccine candidates (PVC) from 37 different pathogens. Our tool predicted 175 PVCs correctly out of 219 sequences. We also tested Vaxi-DL on different datasets obtained from multiple resources. Our tool has demonstrated an average sensitivity of 93% and will thus be a useful tool for prioritising PVCs for preclinical studies.


Genetic modification of human embryonic stem cells for derivation of target cells.
Directed differentiation of human embryonic stem cells (hESCs) may yield models to study organogenesis, produce cells and tissues for therapies, and identify clinically relevant compounds for disease treatment. Optimal conditions for specific differentiation of hESCs are still being determined. Incorporation of fluorescent reporter genes will enable high-throughput screening to identify fate-specifying molecules. Ectopic expression, or silencing, of key developmental genes can also direct differentiation toward specific lineages. Here, we briefly overview various genetic modifications used to generate useful hESC lines. We identify strengths and limitations to each method and propose the most suitable approaches for different applications.


Aerobic scope in chicken embryos.
We investigated the aerobic scope of chicken embryos, that is, the margin of increase of oxygen consumption ( [Formula: see text] ) above its normal value. [Formula: see text] was measured by an open-flow methodology at embryonic ages E3, E7, E11, E15, E19 and at E20 at the internal (IP) and external pipping (EP) phases, at the normal incubation temperature (Ta=38°C), in hypothermia (Ta=30°C) and in hyperthermia (Ta=41 and 44°C). In the cold, Q10 averaged ~2 at all ages, except in IP and EP when lower values (~1.5) indicated some degree of thermogenesis. In hyperthermia (38-44°C) Q10 was between 1 and 1.4. Hyperthermia had no significant effects on [Formula: see text] whether the results combined all ages or considered individual age groups, except in IP (in which [Formula: see text] increased 8% with 44°C) and EP embryos (+13%). After opening the air cell, which exposed the embryo to a higher O2 pressure, hyperthermic [Formula: see text] was significantly higher than in normothermia in E19 (+13%), IP (+22%) and EP embryos (+22%). We conclude that in chicken embryos throughout most of incubation neither heat nor oxygen availability limits the normal (normoxic-normothermic) values of [Formula: see text] . Only close to hatching O2-diffusion represents a limiting factor to the embryo's [Formula: see text] . Hence, embryos differ from postnatal animals for a nearly absent aerobic scope, presumably because their major sources of energy expenditure (growth and tissue maintenance) are constantly maximized.


Optical diffraction analysis of electron microscope images of tubular structures found in cells infected with herpes simplex virus type 2.
Optical diffraction analysis was done on the electron micrographs of tubular structures found in cells infected with herpes simplex virus type 2. By this method, the helical arrangement of subunits forming a tubular structure was demonstrated.


Utilization of cyanopyridine in design and synthesis of first-in-class anticancer dual acting PIM-1 kinase/HDAC inhibitors.
Herein, we report design and synthesis of twenty-one dual PIM-1/HDAC inhibitors utilizing 3-cyanopyridines as a novel cap moiety linked with aliphatic /aromatic linker bearing carboxylic acid 3a-g, hydroxamic acid 4a-g or 2-aminoanilide moieties 5a-g as zinc-binding group. Most of the target hybrids revealed promising growth inhibition according to one dose NCI protocol against 60 cancer cell lines. Meanwhile, hydroxamic acids 4b, 4d and 4e displayed strong and broad-spectrum activity against nine tumor subpanels tested (GI50 0.176-8.87 μM); 4d displayed strong antiproliferative activity with GI50 ≤ 3 μM against different cancer cell lines (GI50 range from 0.325 to 2.9 μM). Furthermore, 4a, 4d-4g and 5f manifested a high inhibitory activity against HDACs 1 and 6 isozymes; 4g, displayed potent HDAC 1 and 6 inhibitory activity (45.01 ± 2.1 and 19.78 ± 1.1 nM) more than the reference SAHA (51.54 ± 2.4 and 21.38 ± 1.2 nM, respectively), while 4f was more potent (30.09 ± 1.4 nM) than SAHA against HDAC 1 and less potent (30.29 ± 1.7 nM) than SAHA against HDAC 6. Hybrids 4b, 4d, 4e and 4f exhibited potent PIM-1 inhibitory activity; 4d showed comparable activity to quercetin (IC50 of 343.87 ± 16.6 and 353.76 ± 17.1 nM, respectively); it exhibited pre G1 apoptosis and arrest cell cycle at G2/M phase. Moreover, it revealed good binding into pocket of HDACs 1,6 and PIM-1 kinase enzymes with good correlation with biological results. Moreover, 4b, 4d and 4e had reasonable drug-likeness properties according to Lipinski's rule. However, multitarget inhibitor of PIM-1/HDAC is a promising strategy in anticancer drug discovery; the most potent hybrids require further in vivo and clinical investigations.


Special stem cells for bone.
Mesenchymal stem cells (MSCs) are multipotential in vitro, but their endogenous properties are poorly defined. In this issue of Cell Stem Cell, Park et al. (2012) report that an MSC-like, osteolineage-directed Mx1+ population generates new osteoblasts at sites of bone damage, suggesting its potential for skeletal repair and regeneration.


[Field 4. Environmental safety practice in the intensive care unit. French-speaking Society of Intensive Care. French Society of Anesthesia and Resuscitation].
Medical devices are known to carry risks from design to scrap. Accident reports in ICU show that medical device account for only 20% of accidents. Formation of users and providing a postmarketing incident reporting are thus essential in health institutions. Clinical and engineering departments should cooperate to produce and secure procedures which should be applied during the lifetime of each clinical device. Several points should be especially fulfilled: close cooperation between clinical departments and biomedical engineering departments with available technicians, computer-based inventory of all devices, evaluation of specifications required before purchasing a new device, education of users on utilisation and maintenance, technical follow-up of devices and keeping maintenance and repair logs, ability to provide users with replacement devices, provision of check-lists before use, forging criteria to decide when device should be discarded. These principles are simple and should be considered as mandatory in order to improve medical device related security.


Upregulation of miR-370 contributes to the progression of gastric carcinoma via suppression of FOXO1.
FOXO1 is downregulated in a number of cancers. However, the underlying mechanisms are poorly understood. In this study, we report that the expression of miR-370 was upregulated in gastric cancer cell lines and gastric cancer tissues. Overexpression of miR-370 in gastric cancer cells promoted the cell proliferation and anchorage-independent growth, while silencing of miR-370 showed opposite effects. miR-370-induced proliferation was correlated with the downregulation of cyclin-dependent kinase inhibitors, p27(Kip1) and p21(Cip1), and the upregulation of the cell cycle regulator cyclin D1. Furthermore, we identified that FOXO1 is the functional target of miR-370. Restored expression of FOXO1 together with miR-370 strongly abrogated miR-370-induced cell proliferation. Taken together, our results revealed a novel mechanism of FOXO1 suppression mediated by miR-370 in gastric cancer.


Photoelectron resonance capture ionization (PERCI): a novel technique for the soft-ionization of organic compounds.
Photoelectron resonance capture ionization (PERCI) is demonstrated as a sensitive ionization technique involving minimal fragmentation of organic molecules. PERCI has been used successfully to softly and efficiently ionize both strongly UV absorbing and non-absorbing molecules. Tunable low energy (<1 eV) electrons are generated by focusing a pulsed UV laser on an aluminum photocathode in the presence of gas phase analyte. Negative ions are formed through a resonance electron capture process. Mass analysis is done using a reflectron time-of-flight mass spectrometer. PERCI is demonstrated for a number of gas phase compounds and simple mixtures, including sulfur hexafluoride, nitrobenzene, nitrophenol, 2-pentanone, hexanal, heptanal, and octanal. In all cases the molecular ion (or [M - H](-)) was observed to be the dominant peak. The 1sigma limit of detection was estimated to be on the order of 10(6) molecules in the ionization region.