[Spatial-temporal variation of the macroinvertebrate epibiont community on red mangrove prop roots Rhizophora mangle (Rhizophoraceae) at La Mancha coastal lagoon, Veracruz, Mexico].
La Mancha lagoon is connected to the Gulf of Mexico through an estuarine ephemeral inlet, a sand bar, which opens during the rainy season and closes during the late fall and the winter storm season. As the lagoon fills up with the permanent stream flow into its Southern part, the water level increases and the sand bar opens up, releasing huge volumes of water into the ocean and leaving the previously flooded intertidal zone exposed to drying. In this study, we described the spatial and temporal variations of structure and composition of the community living on the surface of red mangrove prop roots, related to the hydrodynamic of the estuarine inlet. Four red mangrove prop roots were collected every three months, over an annual period, at ten sampling stations. Water temperature, salinity, dissolved oxygen, water depth and transparency were also measured at each sampling event. To describe the identity, abundance, biomass, and specific richness of epibiont coverage on the prop roots, measurements were taken at 10 cm intervals (herein called "horizons"). A total of 59,632 specimens and a total biomass of 172 g/cm2 were registered, and a maximum of eleven horizons presented at least one" species of epibionts. Two species of polychaetes, 12 of mollusks and 14 of crustaceans were identified, with Mytilopsis leucophaeata as the most abundant, Ficopomatus miamiensis as the most common and Crassostrea rhizophorae with the highest contribution in biomass. Sampling effort was assessed using species accumulation curves, resulting that the efficiency was, in general, greater than 90% of the richness predicted by asymptotic models. The hydrodynamics of the estuarine inlet had a great influence on the environmental conditions of the lagoon and on the spatial and temporal variation of the epibiont community: (a) when the inlet is open, there is a North-South gradient in environmental parameters (b) when the inlet is closed and the water level increases, the epibionts especially the pioneer species F. miamiensis, occupied the highest horizons of prop roots, and (c) the abundance, biomass and richness of epibionts increased towards the deeper and less saline Southern part of the lagoon, as well as on permanently submerged horizons, and during the seasonal closure of the inlet. Variations in depth are determined by the hydrodynamic behavior of the estuarine-inlet, suggesting that these factors are major biodiversity drivers of the epibiont community at La Mancha.


Age determination of burned flint by a thermoluminescent method.
Interference by tribothermoluminescence and by regeneration thermoluminescence can be eliminated for burned flint if thin polished slices are used instead of the powders conventionally used in thermoluminescent measurements. Measurement of the internal and external radiation dose rates then makes possible the absolute determination of the date of pyrolization of small fragments of chert or chalcedony to an accuracy of about 10 to 15 percent. The method may be usable to ages in the range from 200,000 to 300,000 years before the present (B.P.). For samples from the archeological site at Carigüela in southern Spain dates from 12,000 to 50,000 years B.P. have been determined, covering deposits ranging from Late Paleolithic-Epipaleolithic to Mousterian. One Bronze Age specimen gave an age of 4,300 years B.P., in excellent agreement with the age determined by other methods.


Effects of feeding dried distillers grains with supplemental starch on fecal shedding of Escherichia coli O157:H7 in experimentally inoculated steers.
Distillers grains (DG), a co-product of ethanol production used as protein and energy supplements in cattle diets, have been shown to increase fecal shedding of Escherichia coli O157:H7, a major foodborne pathogen. The reason for the positive association is not known. Because DG often replaces grain in the diet, decreased starch content and flow to the hindgut may create a favorable environment for E. coli O157:H7. Our objective was to determine whether the addition of starch to a corn DG-supplemented diet negates the effects of DG on fecal shedding of E. coli O157:H7. We conducted a study with 21 steers fed 1 of 3 diets: a corn grain-based basal diet (CON), basal diet supplemented with 25% corn dried DG (DDG), and basal diet supplemented with 25% DDG with corn starch (DDG+S) added at a level intended to increase starch concentration to that of the CON diet. Steers, housed individually in a biosafety level 2 animal facility, were randomly allocated to treatment diets and orally inoculated with a 5-strain mixture (10(9) cfu per steer) of nalidixic acid-resistant (Nal(R)) E. coli O157:H7. Fecal samples were collected for 5 wk, and on d 35, steers were euthanized and necropsied to collect gut content samples. Fecal or gut samples were cultured to determine prevalence and concentrations of Nal(R) E. coli O157:H7. Dietary starch concentrations, based on feed analysis, were 46.3% in the CON compared with 43.3 and 41.3% in the DDG and DDG+S diets, respectively. Steers fed DDG or DDG+S diets shed Nal(R) E. coli O157:H7 more often (P = 0.0027 and 0.0003, respectively) and at greater concentrations (1.9 or 2.0 cfu/g; P = 0.0025 and 0.0006) than those fed CON diet (1.4 cfu/g), but no difference was observed between DDG and DDG+S diets. Cumulative prevalence and concentrations of Nal(R) E. coli O157:H7 were greater in gut samples collected at necropsy in steers fed DDG and DDG+S diets compared with those fed CON diet, but no difference was observed between DDG and DDG+S diets. The lack of effect of starch addition to the DDG diet on fecal shedding of E. coli O157:H7 may be because either the decreased starch content in the DG-supplemented diet is not a factor in the increased shedding of E. coli O157:H7 or inclusion of pure starch in the diet may not have achieved our intended goal to have starch flow into the hindgut similar to that of corn grain. The study confirms our previous finding of the positive association between feeding DG and fecal shedding of E. coli O157:H7.


Use of scanning electron microscopy to investigate the prophylactic efficacy of rifampin-impregnated CSF shunt catheters.
Infection continues to be one of the major complications of cerebrospinal fluid (CSF) shunting procedures, and is caused mainly by skin-derived bacteria. Production of an extracellular biofilm plays an important role in the pathogenesis of shunt-associated infections by protecting bacteria from immune mechanisms and antibiotics. So far, removal of the original shunt and implantation of a new shunting device has been the only successful treatment for most patients. As an alternative strategy to prevent CSF infections, a rifampin-impregnated silicone catheter was designed to provide high initial and long-lasting (>60 days) release of bactericidal drug. To investigate the pathophysiological mechanism of its function, this new device was investigated both in vitro and in a rodent model of CSF infection by scanning electron microscopy (SEM) and bacterial culture. Staphylococcus epidermidis (10(8) cfu/ml) and S. aureus (10(4) cfu/ml) served as test strains. SEM demonstrated that, in contrast to the unloaded catheters, initial bacterial adherence on the catheter surface could be reduced to a few single cells, which did not show visible signs of proliferation. Bacterial cultures obtained simultaneously were all sterile, showing that adherent bacteria were killed immediately by the rifampin released from the catheter. Although rifampin incorporation into silicone polymers was not able to prevent initial bacterial adhesion completely, subsequent colonisation could be prevented.


All-trans-retinoic acid promotes trafficking of human concentrative nucleoside transporter-3 (hCNT3) to the plasma membrane by a TGF-beta1-mediated mechanism.
Human concentrative nucleoside transporter-3 (hCNT3) is a sodium-coupled nucleoside transporter that exhibits high affinity and broad substrate selectivity, making it the most suitable candidate for mediating the uptake and cytotoxic action of most nucleoside-derived drugs. The drug of this class most commonly used in the treatment of chronic lymphocytic leukemia (CLL) is the pro-apoptotic nucleoside analog fludarabine (Flu), which enters CLL cells primarily through human equilibrative nucleoside transporters (hENTs). Although CLL cells lack hCNT3 activity, they do express this transporter protein, which is located mostly in the cytosol. The aim of our study was to identify agents and mechanisms capable of promoting hCNT3 trafficking to the plasma membrane. Here, we report that all-trans-retinoic acid (ATRA), currently used in the treatment of acute promyelocytic leukemia (APL), increases hCNT3-related activity through a mechanism that involves trafficking of pre-existing hCNT3 proteins to the plasma membrane. This effect is mediated by the autocrine action of transforming growth factor (TGF)-beta1, which is transcriptionally activated by ATRA in a p38-dependent manner. TGF-beta1 acts through activation of ERK1/2 and the small GTPase RhoA to promote plasma membrane trafficking of the hCNT3 protein.


Crystal engineering of zeolites with graphene.
Achieving control over the morphology of zeolite crystals at the nanoscale is crucial for enhancing their performance in diverse applications including catalysis, sensors and separation. The complexity and sensitivity of zeolite synthesis processes, however, often make such control both highly empirical and difficult to implement. We demonstrate that graphene can significantly alter the morphology of titanium silicalite (TS-1) particles, in particular being able to reduce their dimensions from several hundreds to less than 10 nm. Through electron microscopy and molecular mechanics simulations we propose a mechanism for this change based on the preferential interaction of specific TS-1 surfaces with benzyl-alcohol-mediated graphene. These findings suggest a facile new means of controlling the zeolite morphology and thereby also further demonstrate the potential of graphene in hybrid materials. Moreover, the generality of the mechanism points the way to a new avenue of research in using two-dimensional materials to engineer functional inorganic crystals.


The Pugwash scientists' conferences, Cyrus Eaton and the clash of internationalisms, 1954-1961.
This paper examines the contest between Canadian American industrialist Cyrus Eaton and the Pugwash scientists' leadership for influence over the early Pugwash scientists' conferences. Eaton's activism has generally been dismissed in the historical literature as ineffective, naive and too uncritical of the Soviet Union. This paper argues that he was genuinely committed to international peace and security, that Eaton shared with Pugwash scientists a belief in the importance of intellectuals to global unity, and that he worked to bring about greater international peace and understanding through both his personal activism and his own conferences held in the town of Pugwash. Eaton, however, favoured a broader push for peace, which included participation by a wider range of intellectuals and a call for rapprochement with the Soviet Union. These differences between Eaton and the Pugwash scientists, I argue, were more than simply about approach: they represented different internationalist visions and manifested in different conceptualizations for the Pugwash scientists' conferences. Eaton hoped to incorporate non-scientists and humanism into the conferences, and integrate them into his own broader conference programme at his Thinkers' Lodge at Pugwash. The scientists, however, wanted to keep their conferences as distinctly scientists' events, tied to science and its authority.


Central-peripheral Temperature Monitoring as a Marker for Diagnosing Late-onset Neonatal Sepsis.
The prognosis for late-onset sepsis depends largely on a timely diagnosis. We assess central-peripheral temperature difference monitoring as a marker for late-onset neonatal sepsis diagnosis. We performed a prospective, observational study focusing on a cohort of 129 very low-birth-weight infants. Thermal gradient alteration was defined as a difference of > 2°C maintained during 4 hours. We then determined its association with the late-onset sepsis variable through logistic regression. We enrolled 129 preterm babies in 52 months. Thermal gradient alterations showed an adjusted odds ratio for late-onset sepsis of 23.60 (95% confidence interval [CI], 6.80-81.88), with a sensitivity of 83% and negative predictive value of 94%. In 71% of cases, thermal gradient alteration was the first clinical sign of sepsis, while C-reactive protein was < 1.5 mg/dL in 64% of cases and procalcitonin < 2 ng/mL in 36%. These figures indicate potential for early diagnosis. Sustained increases of central-peripheral temperature differences are an early sign of evolving late-onset sepsis.


Arabidopsis CAP1 - a key regulator of actin organisation and development.
Maintenance of F-actin turnover is essential for plant cell morphogenesis. Actin-binding protein mutants reveal that plants place emphasis on particular aspects of actin biochemistry distinct from animals and fungi. Here we show that mutants in CAP1, an A. thaliana member of the cyclase-associated protein family, display a phenotype that establishes CAP1 as a fundamental facilitator of actin dynamics over a wide range of plant tissues. Plants homozygous for cap1 alleles show a reduction in stature and morphogenetic disruption of multiple cell types. Pollen grains exhibit reduced germination efficiency, and cap1 pollen tubes and root hairs grow at a decreased rate and to a reduced length. Live cell imaging of growing root hairs reveals actin filament disruption and cytoplasmic disorganisation in the tip growth zone. Mutant cap1 alleles also show synthetic phenotypes when combined with mutants of the Arp2/3 complex pathway, which further suggests a contribution of CAP1 to in planta actin dynamics. In yeast, CAP interacts with adenylate cyclase in a Ras signalling cascade; but plants do not have Ras. Surprisingly, cap1 plants show disruption in plant signalling pathways required for co-ordinated organ expansion suggesting that plant CAP has evolved to attain plant-specific signalling functions.


Cantilever tip near-field surface-enhanced Raman imaging of tris(bipyridine)ruthenium(II) on silver nanoparticles-coated substrates.
The near-field surface-enhanced Raman scattering (SERS) and surface-enhanced fluorescence (SEF) images of tris(bipyridine)ruthenium(II) adsorbed on a silver nanoparticles-coated substrate were obtained with a scanning near-field optical microscope (SNOM, or near-field scanning optical microscopy, NSOM) using a cantilever tip. In comparison with the most widely used fiber tip for SNOM, the cantilever tip has higher optical throughput and better thermal stability, making it more suitable for detecting the extremely low Raman signal in the near-field spectroscopic investigations. Our preliminary results show that the near-field SERS with the higher spatial resolution can provide richer fingerprint information than the far-field SERS. A comparison of the two types of images shows that there are more SERS than SEF hot spots, and the two types of hot spots do not overlap. More surprisingly, the near-field SERS spectra differ from the far-field SERS spectra obtained on the same sample in the band frequency and relative intensities of some major Raman bands, and some IR-active bands were observed with the near-field mode. These results are explained mainly by the electric field gradient effect and heterogeneous polarization character that operate only in the near-field SERS.


Vasculogenic mimicry.
The term vasculogenic mimicry describes the formation of fluid-conducting channels by highly invasive and genetically dysregulated tumor cells. Two distinctive types of vasculogenic mimicry have been described. Vasculogenic mimicry of the tubular type may be confused morphologically with endothelial cell-lined blood vessels. Vasculogenic mimicry of the patterned matrix type in no way resembles blood vessels morphologically or topologically. Matrix proteins such as laminin, heparan sulfate proteoglycan, and collagens IV and VI have been identified in these patterns. The patterned matrix anastomoses with blood vessels, and systemically injected tracers co-localize to these patterns. Vasculogenic mimicry of the patterned matrix type has been identified in uveal, cutaneous and mucous membrane melanomas, inflammatory and ductal breast carcinoma, ovarian and prostatic carcinoma, and soft tissue sarcomas, including synovial sarcoma rhabdomyosarcoma, osteosarcoma, and pheochromocytoma. Because the microcirculation of many tumors may be heterogeneous -- including incorporated or co-opted vessels, angiogenic vessels, mosaic vessels, and vasculogenic mimicry of the tubular and patterned matrix types -- therapeutic regimens that target angiogenesis alone may be ineffective against highly invasive tumors that contain patterned matrices. Vasculogenic mimicry provides an opportunity to investigate the interrelationships between the genetically dysregulated invasive tumor cell, the microenvironment, and the malignant switch.


[Treatment of stage I-IIA breast cancer of medial-central localization].
702 patients with breast cancer were followed-up after, sparing and radical operations combined with radiation and polychemotherapy. Of adjuvant treatments, radiation therapy showed to be more effective than chemotherapy. Methods of operative treatment combined with radiation are not essential in the treatment of early stages of breast cancer.


Microsomal prostaglandin synthase-1-derived prostaglandin E2 protects against angiotensin II-induced hypertension via inhibition of oxidative stress.
Prostaglandin (PG) E(2) has an established role in the regulation of vascular tone and reactivity. The present study examined the role and mechanism of microsomal PG synthase-1 (mPGES-1) in vascular response to angiotensin II (Ang II) infusion. A 7-day Ang II infusion at 0.35 mg/kg per day via osmotic minipump had no obvious effect on mean arterial blood pressure in mPGES-1(+/+) mice but induced a marked hypertensive response in mPGES-1(-/-) mice, associated with a parallel increase in urinary 8-isoprostane excretion and aortic NADPH oxidase activity and mRNA expression of p47(phox), gp91(phox), and Nox1. The hypertension in mPGES-1(-/-) mice was completely prevented by Tempol treatment and was fully restored on termination of the antioxidant. Apocynin induced a similar blood pressure-lowering effect as Tempol. The Ang II infusion induced mRNA expression of mPGES-1, as well as mPGES-2 and cytosolic PGE synthase in the aortas as assessed by real-time RT-PCR. Immunohistochemistry revealed remarkably enhanced immunoreactivity of mPGES-1 mostly in vascular smooth muscle cells. In cultured vascular smooth muscle cells, Ang II exerted a direct stimulatory effect on reactive oxygen species production, NADPH oxidase activity, and expression of p47(phox), gp91(phox), and Nox1 that were all inhibited by PGE(2). The -/- mice also exhibited enhanced renal hemodynamic response to acute Ang II infusion at 150 nmol/kg per minute via a jugular vein over a period of 40 minutes. These results suggest that mPGES-1-derived PGE(2) buffers Ang II-induced vasoconstriction via inhibition of NADPH oxidase-dependent reactive oxygen species production.


Diabetes and Obesity in Down Syndrome Across the Lifespan: A Retrospective Cohort Study Using U.K. Electronic Health Records.
Down syndrome (DS) is the most common form of chromosomal trisomy. Genetic factors in DS may increase the risk for diabetes. This study aimed to determine whether DS is associated with an increased incidence of diabetes and the relationship with obesity across the life span compared with control patients. This matched population-based cohort study analyzed UK Clinical Practice Research Datalink data from 1990 to 2020. A total of 9,917 patients with DS and 38,266 control patients were analyzed. Diabetes rates were higher in patients with DS (incidence rate ratio 3.67; 95% CI 2.43-5.55; P < 0.0001) and peaked at a younger age (median age at diagnosis 38 [interquartile range 28-49] years vs. 53 [43-61] years in control patients). Incidence rates (per 1,000 person-years) for type 1 diabetes mellitus were 0.44 (95% CI 0.31-0.61) in patients with DS vs. 0.13 (0.09-0.17) in control patients. Type 2 diabetes mellitus (T2DM) rates were higher in patients with DS versus control patients in age-groups from 5 years up to 34 years. In patients with DS, peak mean BMI was higher and at a younger age (males 31.2 kg/m2 at age 31 years; females 32.1 kg/m2 at 43 years) versus control patients (males 29.5 kg/m2 at 54 years; females 29.2 kg/m2 at 51 years). Obesity was associated with an increased incidence of T2DM. At younger ages, the incidence of diabetes in patients with DS is up to four times that of control patients. Peak mean BMI is higher and established earlier in DS, contributing to T2DM risk. Further investigation into the relationship between obesity and diabetes in DS is required to inform treatment and prevention measures.


Rapid phenotypic change and diversification of a soil bacterium during 1000 generations of experimental evolution.
Evolutionary pathways open to even relatively simple organisms, such as bacteria, may lead to complex and unpredictable phenotypic changes, both adaptive and non-adaptive. The evolutionary pathways taken by 18 populations of Ralstonia strain TFD41 while they evolved in defined environments for 1000 generations were examined. Twelve populations evolved in liquid media, while six others evolved on agar surfaces. Phenotypic analyses of these derived populations identified some changes that were consistent across all populations and others that differed among them. The evolved populations all exhibited morphological changes in their cell envelopes, including reductions of the capsule in each population and reduced prostheca-like surface structures in most populations. Mean cell length increased in most populations (in one case by more than fourfold), although a few populations evolved shorter cells. Carbon utilization profiles were variable among the evolved populations, but two distinct patterns were correlated with genetic markers introduced at the outset of the experiment. Fatty acid methyl ester composition was less variable across populations, but distinct patterns were correlated with the two physical environments. All 18 populations evolved greatly increased sensitivity to bile salts, and all but one had increased adhesion to sand; both patterns consistent with changes in the outer envelope. This phenotypic diversity contrasts with the fairly uniform increases in competitive fitness observed in all populations. This diversity may represent a set of equally probable adaptive solutions to the selective environment; it may also arise from the chance fixation of non-adaptive mutations that hitchhiked with a more limited set of beneficial mutations.


Molecular cloning, characterization, and chromosomal localization of dapF, the Escherichia coli gene for diaminopimelate epimerase.
The Escherichia coli dapF gene was isolated from a cosmid library as a result of screening for clones overproducing diaminopimelate epimerase. Insertional mutagenesis was performed on the cloned dapF gene with a mini-Mu transposon, leading to chloramphenicol resistance. One of these insertions was transferred onto the chromosome by a double-recombination event, allowing us to obtain a dapF mutant. This mutant accumulated large amounts of LL-diaminopimelate, confirming the blockage in the step catalyzed by the dapF product, but did not require meso-diaminopimelate for growth. The dapF gene was localized in the 85-min region of the E. coli chromosome between cya and uvrD.


The Role of Coronary Artery Calcium Testing for Value-Based Clinical Trials in Primary Prevention.
Review the role of coronary artery calcium (CAC) testing in designing future clinical trials in primary prevention. While there are numerous new agents that have been found to lower cardiovascular event rates in clinical trials, these studies have required a large sample size, in part due to low event rates as well as improved baseline treatments. More precise risk assessment could allow for better identification of individuals who stand to derive the most benefit from various therapies. Coronary CAC testing offers a simple method for identifying high-risk primary prevention cohorts, and thus may allow for improved efficiency of clinical trials, enhanced efficacy of various therapies, and ultimately more favorable cost-effectiveness estimates. The use of CAC testing as part of the inclusion criteria used in clinical trials may result in identifying high-risk individuals who were previously not included in such studies while achieving favorable absolute risk reductions. The advantages afforded by using CAC to enrich clinical trials offer a potential road map for future clinical trials in primary prevention.


Maternal intravenous administration of azithromycin results in significant fetal uptake in a sheep model of second trimester pregnancy.
Treatment of intrauterine infection is likely key to preventing a significant proportion of preterm deliveries before 32 weeks of gestation. Azithromycin (AZ) may be an effective antimicrobial in pregnancy; however, few gestation age-approriate data are available to inform the design of AZ-based treatment regimens in early pregnancy. We aimed to determine whether a single intra-amniotic AZ dose or repeated maternal intravenous (i.v.) AZ doses would safely yield therapeutic levels of AZ in an 80-day-gestation (term is 150 days) ovine fetus. Fifty sheep carrying single pregnancies at 80 days gestation were randomized to receive either: (i) a single intra-amniotic AZ administration or (ii) maternal intravenous AZ administration every 12 h. Amniotic fluid, maternal plasma, and fetal AZ concentrations were determined over a 5-day treatment regimen. Markers of liver injury and amniotic fluid inflammation were measured to assess fetal injury in response to drug exposure. A single intra-amniotic administration yielded significant AZ accumulation in the amniotic fluid and fetal lung. In contrast, repeated maternal intravenous administrations achieved high levels of AZ accumulation in the fetal lung and liver and a statistically significant increase in the fetal plasma drug concentration at 120 h. There was no evidence of fetal injury in response to drug exposure. These data suggest that (i) repeated maternal i.v. AZ dosing yields substantial fetal tissue uptake, although fetal plasma drug levels remain low; (ii) transfer of AZ from the amniotic fluid is less than transplacental transfer; and (iii) exposure to high concentrations of AZ did not elicit overt changes in fetal white blood cell counts, amniotic fluid monocyte chemoattractant protein 1 concentrations, or hepatotoxicity, all consistent with an absence of fetal injury.


Cognitive-behaviour therapy for post-traumatic stress in schizophrenia. A randomized controlled trial.
There is limited evidence for effective interventions in the treatment of post-traumatic stress symptoms within individuals diagnosed with schizophrenia. Clinicians have concerns about using exposure treatments with this patient group. The current trial was designed to evaluate a 16-session cognitive restructuring programme, without direct exposure, for the treatment of post-traumatic stress symptoms specifically within individuals diagnosed with schizophrenia. A multicentre randomized controlled single-blinded trial with assessments at 0 months, 6 months (post-treatment) and 12 months (follow-up) was conducted. A total of 61 participants diagnosed with schizophrenia and exhibiting post-traumatic stress symptoms were recruited. Those randomized to treatment were offered up to 16 sessions of cognitive-behaviour therapy (CBT, including psychoeducation, breathing training and cognitive restructuring) over a 6-month period, with the control group offered routine clinical services. The main outcome was blind rating of post-traumatic stress symptoms using the Clinician Administered PTSD Scale for Schizophrenia. Secondary outcomes were psychotic symptoms as measured by the Positive and Negative Symptom Scale and the Psychotic Symptom Rating Scale. Both the treatment and control groups experienced a significant decrease in post-traumatic stress symptoms over time but there was no effect of the addition of CBT on either the primary or secondary outcomes. The current trial did not demonstrate any effect in favour of CBT. Cognitive restructuring programmes may require further adaptation to promote emotional processing of traumatic memories within people diagnosed with a psychotic disorder.


Modeling behavior in different delay match to sample tasks in one simple network.
Delay match to sample (DMS) experiments provide an important link between the theory of recurrent network models and behavior and neural recordings. We define a simple recurrent network of binary neurons with stochastic neural dynamics and Hebbian synaptic learning. Most DMS experiments involve heavily learned images, and in this setting we propose a readout mechanism for match occurrence based on a smaller increment in overall network activity when the matched pattern is already in working memory, and a reset mechanism to clear memory from stimuli of previous trials using random network activity. Simulations show that this model accounts for a wide range of variations on the original DMS tasks, including ABBA tasks with distractors, and more general repetition detection tasks with both learned and novel images. The differences in network settings required for different tasks derive from easily defined changes in the levels of noise and inhibition. The same models can also explain experiments involving repetition detection with novel images, although in this case the readout mechanism for match is based on higher overall network activity. The models give rise to interesting predictions that may be tested in neural recordings.


A Randomized Controlled Trial of Two Semi-Occluded Vocal Tract Voice Therapy Protocols.
Although there is a long history of use of semi-occluded vocal tract gestures in voice therapy, including phonation through thin tubes or straws, the efficacy of phonation through tubes has not been established. This study compares results from a therapy program on the basis of phonation through a flow-resistant tube (FRT) with Vocal Function Exercises (VFE), an established set of exercises that utilize oral semi-occlusions. Twenty subjects (16 women, 4 men) with dysphonia and/or vocal fatigue were randomly assigned to 1 of 4 treatment conditions: (a) immediate FRT therapy, (b) immediate VFE therapy, (c) delayed FRT therapy, or (d) delayed VFE therapy. Subjects receiving delayed therapy served as a no-treatment control group. Voice Handicap Index (Jacobson et al., 1997) scores showed significant improvement for both treatment groups relative to the no-treatment group. Comparison of the effect sizes suggests FRT therapy is noninferior to VFE in terms of reduction in Voice Handicap Index scores. Significant reductions in Roughness on the Consensus Auditory-Perceptual Evaluation of Voice (Kempster, Gerratt, Verdolini Abbott, Barkmeier-Kraemer, & Hillman, 2009) were found for the FRT subjects, with no other significant voice quality findings. VFE and FRT therapy may improve voice quality of life in some individuals with dysphonia. FRT therapy was noninferior to VFE in improving voice quality of life in this study.


Impact of TP53 codon 72 and MDM2 SNP 309 polymorphisms in pancreatic ductal adenocarcinoma.
Single-nucleotide polymorphisms (SNPs) of TP53 (codon 72, rs1042522) and MDM2 promoter (SNP 309, rs2279744) have been associated with risk for various human cancers. However, studies analyzing these polymorphisms in pancreatic ductal adenocarcinoma (PDAC) are lacking. We investigated TP53 codon 72 and MDM2 SNP 309 polymorphisms in 32 patients with PDAC, 16 patients with chronic pancreatitis (CP), and 32 normal controls, using formalin-fixed paraffin-embedded tissue. We also examined TP53 and MDM2 protein immunohistochemistry (IHC) to assess the involvement of these differences in malignant transformation and disease progression. TP53 Pro/Pro genotype was significantly more frequent in PDAC patients than in controls (65.6 vs. 15.6%, p < 0.001) and no significant difference was found between CP patients (37.5%) and controls. In MDM2 SNP 309, there were no significant differences among the three groups. Based on the Kaplan-Meier analysis, overall survival was significantly shorter in MDM2 G/G genotypes compared with other genotypes (G/T and T/T) (359 vs. 911 days, p = 0.016) whereas no significant differences in TP53 genotypes were observed (638 vs. 752 days, p = 0.471). Although TP53 IHC was frequent in PDAC patients (53.1%), TP53 and MDM2 protein expression was not correlated with polymorphisms. Our study demonstrated TP53 codon 72 polymorphism is potentially a genetic predisposing factor while MDM2 SNP 309 polymorphism might be useful in predicting survival outcome.


Influence of growth rate on the physiological response of marine Synechococcus to phosphate limitation.
Phosphate (P) is an important nutrient potentially limiting for primary productivity, yet, we currently know little about the relationship between growth rate and physiological response to P limitation in abundant marine Cyanobacteria. Thus, the aim of this research was to determine how variation in growth rate affected the physiology of marine Synechococcus WH8102 and CC9311 when growing under high N:P conditions. Experiments were carried out in chemostats with a media input N:P of 441 and we estimated the half saturation concentration for growth under P limiting conditions (K s,p ) and cellular C:N:P ratios. The K s,p values were the lowest measured for any phytoplankton and on par with ambient P concentrations in oligotrophic regions. We also observed that both strains were able draw down P below 3 nM. Both K s,p and drawdown concentration were lower for the open ocean vs. coastal Synechococcus strain, which may be linked to differences in P acquisition genes in these strains. Cellular C:P and N:P ratios were significantly higher in relation to the Redfield ratio for both Synechococcus strains but we saw no difference in these ratios among growth rates or strains. These results demonstrate that Synechococcus can proliferate under very low P conditions and also that genetically different strains have unique physiological responses to P limitation.


Dew harvesting efficiency of four species of cacti.
Four species of cacti were chosen for this study: Copiapoa cinerea var. haseltoniana, Ferocactus wislizenii, Mammillaria columbiana subsp. yucatanensis and Parodia mammulosa. It has been reported that dew condenses on the spines of C. cinerea and that it does not on the spines of F. wislizenii, and our preliminary observations of M. columbiana and P. mammulosa revealed a potential for collecting dew water. This study found all four cacti to harvest dew on their stems and spines (albeit rarely on the spines of F. wislizenii). Dew harvesting experiments were carried out in the UK, recording an increase in cacti mass on dewy nights. By applying a ranking relative to a polymethyl methacrylate (Plexiglas) reference plate located nearby, it was found that C. cinerea collected the most airborne moisture followed by M. columbiana, P. mammulosa and F. wislizenii respectively, with mean efficiency ratio with respect to the Plexiglas reference of 3.48 ± 0.5, 2.44 ± 0.06, 1.81 ± 0.14 and 1.27 ± 0.49 on observed dewy nights. A maximum yield of normalized performance of 0.72 ± 0.006 l/m(-2) on one dewy night was recorded for C. cinerea. Removing the spines from M. columbiana was found to significantly decrease its dew harvesting efficiency. The spines of three of the species were found to be hydrophilic in nature, while F. wislizenii was hydrophobic; the stems of all four species were hydrophilic. The results of this study could be translated into designing a biomimetic water collecting device that utilizes cactus spines and their microstructures.


Activity of aaptamine and two derivatives, demethyloxyaaptamine and isoaaptamine, in cisplatin-resistant germ cell cancer.
We analyzed the effects of all three marine alkaloids aaptamine, demethyloxyaaptamine and isoaaptamine in NT2-R, a cisplatin-resistant subline of the human embryonal carcinoma cell line NT2. All aaptamines were found to be equally effective in both cell lines, excluding cross-resistance between aaptamines and cisplatin in vitro. At the inhibitory concentration (IC50), aaptamine exerted an antiproliferative effect, whereas demethyloxyaaptamine and isoaaptamine were strong inducers of apoptosis. We analyzed the changes in the proteome of NT2-R cells treated with these compounds. 16-22 proteins were found to be significantly altered, of which several were validated by Western blotting and two-dimensional Western blotting analysis. Changes in the proteome pattern frequently resulted from post-transcriptional protein modifications, i.e. phosphorylation or hypusination in the case of eIF5A. Although the lists of altered proteins were heterogeneous and compound-specific, gene ontology analyses identified rather similar profiles regarding the affected molecular functions. Ingenuity pathway analysis by IPA put the following factors in a central position of the hypothetical networks: myc and p53 for aaptamine; tumor necrosis factor (TNF) for demethyloxyaaptamine; and all three, myc, p53, and TNF for isoaaptamine. Our results represent an important step towards a better understanding of the molecular basis underlying the observed bioactivity of these promising marine compounds. We characterized the mode of action of three aaptamines, marine natural compound with anti-tumor activity, using a functional proteomics approach and the cisplatin-resistant pluripotent human embryonal carcinoma cell line NT2-R. The manuscript is of particular scientific interest, as we could reveal the similarities and differences of the modes of action. Furthermore, we were able to identify several new targets of these promising compounds. We found hypusination of eIF5A to be a prominent feature exclusively of aaptamine treatment, as this was not observed upon treatment with demethyloxyaaptamine or isoaaptamine. Our results are a step towards unraveling the mode of action of these interesting compounds.


New biotechnologies: spin-off on health and society.
New biotechnologies and the new biology deriving from them are having a revolutionary impact on economy and society and are consequently transforming the role of researchers, which is changing continually to meet the competence required. The advances in human genetics on the other hand make it necessary to look for different approaches and new rules in bioethics. Comprehensive views and careful considerations are therefore needed in order that this new biology may have a positive impact on health, being respectful of the social and ethical principles of human beings.


The impact of drop-out in cardiac rehabilitation on outcome among coronary artery disease patients.
Background The effect of adherence to cardiac rehabilitation (CR) on outcome is not clear. Therefore, we aimed to assess the impact of drop-out for non-medical reasons of CR on event-free survival in coronary artery disease (CAD). Methods A total of 876 patients who attended CR after acute coronary syndrome (ACS), percutaneous coronary intervention (PCI) or coronary artery bypass graft (CABG) were included. Drop-out was defined as attending ≤50% of the training sessions. A combined endpoint of all-cause mortality and rehospitalization for a cardiovascular event was used to specify event-free survival. Differences in clinical characteristics were assessed and parameters with p < 0.10 were entered in a multiple Cox regression analysis. Results A total of 15% died or had a cardiovascular event during a median follow-up period of 33 months (interquartile range 24, 51). Overall, 17% dropped out before finishing half of the program. Patients who withdrew prematurely had a risk twice as high for a cardiovascular event or death (hazard ratio 1.92, 95% confidence interval 1.28-2.90) than those who attended more than half of the sessions. Both ACS (2.36, 1.47-3.58) and PCI (2.20, 1.22-3.96), as primary indicators for CR, were associated with an adverse outcome and also a prior history of chronic heart failure (CHF) remained negatively associated with event-free survival (3.67, 1.24-10.91). Finally, the presence of hyperlipidemia was independently related to a worse outcome (1.48, 1.02-2.16). Conclusions Drop-out for non-medical reasons was independently associated with a negative outcome in CAD. Therefore, underlying factors for drop-out should gain more attention in future research and should be taken into account when organizing CR.


Influence of stimulus frequency and probe size on vibration-induced alleviation of acute orofacial pain.
The pain-relieving effect of vibratory stimulation, using different stimulus parameters, and placebo stimulation in acute orofacial pain is reported. The influence of 10-, 100-, and 200-Hz vibrations on pain reduction was studied in 96 patients; two different probe sizes were used. 54 out of 76 patients, receiving vibrations at any of the above frequencies, reported relief of pain to some extent, while only 6 out of 20 patients receiving placebo treatment experienced pain alleviation. No significant differences were found between the different frequencies and probe sizes used regarding the pain-relieving effect. However, placebo stimulation was significantly less effective than any kind of vibratory stimulation. Induction time for pain relief was significantly shorter using the larger probe as compared to using the smaller probe, regardless of frequency. The results indicate that the vibratory frequency (10-200 Hz) for activation of pain-inhibitory mechanisms is not critical in acute orofacial pain. Also, spatial summation from vibration-sensitive afferents seems to be of importance for a fast activation of the inhibitory systems.


Effects of Retzius sparing on robot-assisted laparoscopic prostatectomy: a systematic review with meta-analysis.
To comprehensively evaluate the efficacy and safety of Retzius sparing (RS) for men undergoing robot-assisted laparoscopic prostatectomy (RARP). We searched four electronic databases and reference lists of relevant studies for eligible research published before March 11, 2019. After quality assessment, eligible studies were synthesized for relevant outcomes, including positive surgical margin (PSM), continence, incontinence, complication, console time, and hospital stay. Two randomized clinical trials and four observational studies were included in this study. Quantitative syntheses revealed significantly higher PSM rates in RS-RARP compared with conventional RARP (c-RARP) (odds ratio [OR] 1.68, p = 0.02). Furthermore, we found significantly higher PSM rates at the anterior site in RS-RARP compared with c-RARP (OR 4.34, p = 0.03) and significantly lower incontinence rates in RS-RARP in the first month (OR 0.30, p < 0.001) and 12th month (OR 0.25, p < 0.001). Our syntheses revealed higher PSM rates in the RS-RARP group, especially in the anterior aspect. However, RS-RARP had superior functional outcome of urinary continence and lower console time than did c-RARP with equivalent complication rates. Thus, we suggest that operators pay more attention to making clear surgical margins if the lesion is in anterior prostate when performing RS-RARP.


Disease-associated polymorphisms in 9p21 are not associated with extreme longevity.
The 9p21 region has been pointed out by the genome-wide association studies as a hot spot for disease-associated variants. Most of the diseases linked with the locus are aging-related conditions, such us cardiovascular disease, diabetes and cancer. Centenarians are known to present a reduced risk and delayed onset for these conditions. Here, we aimed to assess if the 9p21 variants contribute to this protection by possibly altering basic aging mechanisms. We genotyped 15 tag single-nucleotide polymorphisms (SNP) along the CDKN2A/B/ANRIL locus in 1505 individuals. The participants were divided in three groups: centenarians, septuagenarians and young controls. Centenarians were 593 participants (age range 100-116 years, mean 105.9 years), septuagenarians were 434 volunteers aged between 69 and 71 years (mean 70.1 ± 0.9 years) and the 478 young controls were under the age of 50 years (range 14-50 years, mean 41.8 years). We genotyped the SNP rs1333049 in an additional sample of 231 coronary artery disease patients to confirm the 9p21 association. The leading coronary artery disease-associated SNP rs1333049 was associated with coronary artery disease; however, none of the 9p21 SNP evaluated in the present study were associated with extreme longevity. Our findings suggest that the 9p21 disease-associated polymorphisms do not contribute to the life-long protection from cardiovascular and other age-related diseases observed in centenarians. It is likely that this protection is mediated by mechanisms different from the ones underlying the 9p21 association.


Context-dependent modulation of hippocampal and cortical recruitment during remote spatial memory retrieval.
According to systems consolidation, as hippocampal-dependent memories mature over time, they become additionally (or exclusively) dependent on extra-hippocampal structures. We assessed the recruitment of hippocampal and cortical structures on remote memory retrieval in a performance-degradation resistant (PDR; no performance degradation with time) versus performance-degradation prone (PDP; performance degraded with time) context. Using a water-maze task in two contexts with a hidden platform and three control conditions (home cage, visible platform with or without access to distal cues), we compared neuronal activation (c-Fos imaging) patterns in the dorsal hippocampus and the medial prefrontal cortex (mPFC) after the retrieval of recent (5 days) versus remote (25 days) spatial memory. In the PDR context, the hippocampus exhibited greater c-Fos protein expression on remote than recent memory retrieval, be it in the visible or hidden platform group. In the PDP context, hippocampal activation increased at the remote time point and only in the hidden platform group. In the anterior cingulate cortex, c-Fos expression was greater for remote than for recent memory retrieval and only in the PDR context. The necessity of the mPFC for remote memory retrieval in the PDR context was confirmed using region-specific lidocaine inactivation, which had no impact on recent memory. Conversely, inactivation of the dorsal hippocampus impaired both recent and remote memory in the PDR context, and only recent memory in the PDP context, in which remote memory performance was degraded. While confirming that neuronal circuits supporting spatial memory consolidation are reorganized in a time-dependent manner, our findings further indicate that mPFC and hippocampus recruitment (i) depends on the content and perhaps the strength of the memory and (ii) may be influenced by the environmental conditions (e.g., cue saliency, complexity) in which memories are initially formed and subsequently recalled.


The SMC5/6 complex is involved in crucial processes during human spermatogenesis.
Genome integrity is crucial for safe reproduction. Therefore, chromatin structure and dynamics should be tightly regulated during germ cell development. Chromatin structure and function are in large part determined by the structural maintenance of chromosomes (SMC) protein complexes, of which SMC5/6 recently has been shown to be involved in both spermatogonial differentiation and meiosis during mouse spermatogenesis. We therefore investigated the role of this complex in human spermatogenesis. We found SMC6 to be expressed in the human testis and present in a subset of type Adark and type Apale spermatogonia, all spermatocytes, and round spermatids. During human meiosis, SMC5/6 is located at the synaptonemal complex (SC), the XY body, and at the centromeres during meiotic metaphases. However, in contrast to mouse spermatogenesis, SMC6 is not located at pericentromeric heterochromatin in human spermatogenic cells, indicating subtle but perhaps important differences in not only SMC5/6 function but maybe also in maintenance of genomic integrity at the repetitive pericentromeric regions. Nonetheless, our data clearly indicate that the SMC5/6 complex, as shown in mice, is involved in numerous crucial processes during human spermatogenesis, such as in spermatogonial development, on the SC between synapsed chromosomes, and in DNA double-strand break repair on unsynapsed chromosomes during pachynema.


Out-of-pocket medication costs and use of medications and health care services among children with asthma.
Health plans have implemented policies to restrain prescription medication spending by shifting costs toward patients. It is unknown how these policies have affected children with chronic illness. To analyze the association of medication cost sharing with medication and hospital services utilization among children with asthma, the most prevalent chronic disease of childhood. Retrospective study of insurance claims for 8834 US children with asthma who initiated asthma control therapy between 1997 and 2007. Using variation in out-of-pocket costs for a fixed "basket" of asthma medications across 37 employers, we estimated multivariate models of asthma medication use, asthma-related hospitalization, and emergency department (ED) visits with respect to out-of-pocket costs and child and family characteristics. Asthma medication use, asthma-related hospitalizations, and ED visits during 1-year follow-up. The mean annual out-of-pocket asthma medication cost was $154 (95% CI, $152-$156) among children aged 5 to 18 years and $151 (95% CI, $148-$153) among those younger than 5 years. Among 5913 children aged 5 to 18 years, filled asthma prescriptions covered a mean of 40.9% of days (95% CI, 40.2%-41.5%). During 1-year follow-up, 121 children (2.1%) had an asthma-related hospitalization and 220 (3.7%) had an ED visit. Among 2921 children younger than 5 years, mean medication use was 46.2% of days (95% CI, 45.2%-47.1%); 136 children (4.7%) had an asthma-related hospitalization and 231 (7.9%) had an ED visit. An increase in out-of-pocket medication costs from the 25th to the 75th percentile was associated with a reduction in adjusted medication use among children aged 5 to 18 years (41.7% [95% CI, 40.7%-42.7%] vs 40.3% [95% CI, 39.4%-41.3%] of days; P = .02) but no change among younger children. Adjusted rates of asthma-related hospitalization were higher for children aged 5 to 18 years in the top quartile of out-of-pocket costs (2.4 [95% CI, 1.9-2.8] hospitalizations per 100 children vs 1.7 [95% CI, 1.3-2.1] per 100 in bottom quartile; P = .004) but not for younger children. Annual adjusted rates of ED use did not vary across out-of-pocket quartiles for either age group. Greater cost sharing for asthma medications was associated with a slight reduction in medication use and higher rates of asthma hospitalization among children aged 5 years or older.


Proteomic profiling of erythrocyte proteins by proteolytic digestion chip and identification using two-dimensional electrospray ionization tandem mass spectrometry.
Self-assembled monolayers (SAMs) on coinage metal provide versatile modeling systems for studies of interfacial electron transfer, biological interactions, molecular recognition, and other interfacial phenomena. The bonding of enzyme to SAMs of alkanethiols onto gold surfaces is exploited to produce an enzyme chip. In this work, the attachment of trypsin to a SAMs surface of 11-mercaptoundecanoic acid was achieved using water soluble N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide as coupling agent. A two-dimensional liquid-phase separation scheme coupled with mass spectrometry is presented for proteomic analysis of erythrocyte proteins. The application of proteomics, particularly with reference to analysis of proteins, will be described. Surface analyses have revealed that the X-ray Photoelectron Spectroscopy (XPS) C1s and N1s core levels illustrate the immobilization of trypsin. These data are also in good agreement with Fourier Transformed Infrared Reflection-Attenuated Total Reflection (FTIR-ATR) spectra for the peaks at Amide I and Amide II. Using two-dimensional nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry (2D nano-HPLC-ESI-MS/MS) system observations, analytical results have demonstrated the erythrocyte proteins digestion of the immobilized trypsin on the functionalized SAMs surface. For such surfaces, it also shows the enzyme digestion ability of the immobilized trypsin. The experiment results revealed the identification of 272 proteins from erythrocyte protein sample. The terminal groups of the SAMs structure can be further functionalized with biomolecules or antibodies to develop surface-base diagnostics, biosensors, or biomaterials.


An Introduction to Infinite HMMs for Single-Molecule Data Analysis.
The hidden Markov model (HMM) has been a workhorse of single-molecule data analysis and is now commonly used as a stand-alone tool in time series analysis or in conjunction with other analysis methods such as tracking. Here, we provide a conceptual introduction to an important generalization of the HMM, which is poised to have a deep impact across the field of biophysics: the infinite HMM (iHMM). As a modeling tool, iHMMs can analyze sequential data without a priori setting a specific number of states as required for the traditional (finite) HMM. Although the current literature on the iHMM is primarily intended for audiences in statistics, the idea is powerful and the iHMM's breadth in applicability outside machine learning and data science warrants a careful exposition. Here, we explain the key ideas underlying the iHMM, with a special emphasis on implementation, and provide a description of a code we are making freely available. In a companion article, we provide an important extension of the iHMM to accommodate complications such as drift.


Witkop tooth and nail syndrome: report of two cases in a family.
Tooth and nail syndrome, an autosomal dominant condition, is characterized by hypodontia and nail dysplasia. Mandibular incisors, second molars and maxillary canines are the most frequently missing teeth. Tooth shape may vary and conical and narrow crowns are common. Nail dysplasia affects fingernails and toenails and is often more severe in childhood. Nails may be spoon-shaped, ridged, slow growing and easily broken. The clinical and radiographic features of a father and son presenting with this rare condition are described.


Mechanistic Evaluation of Hydration Effects on the Human Epidermal Permeation of Salicylate Esters.
We sought to understand when and how hydration enhances the percutaneous absorption of salicylate esters. Human epidermal membrane fluxes and stratum corneum solubilities of neat and diluted solutions of three esters were determined under hydrated and dehydrated conditions. Hydration doubled the human epidermal flux seen for methyl and ethyl salicylate under dehydrated conditions and increased the flux of neat glycol salicylate 10-fold. Mechanistic analyses showed that this hydration-induced enhancement arises mainly from an increase in the stratum corneum diffusivity of the three esters. Further, we showed that unlike methyl and ethyl salicylate, glycol salicylate is hygroscopic and the ∼10-fold hydration-induced flux enhancement seen with neat glycol salicylate may be due to its ability to hydrate the stratum corneum to a greater extent. The hydration-induced enhancements in in vitro epidermal flux seen here for glycol and ethyl salicylate were similar to those reported for their percutaneous absorption rates in a comparable in vivo study, whilst somewhat higher enhancement was seen for methyl salicylate in vivo. This may be explained by a physiologically induced self enhancement of neat methyl salicylate absorption in vivo which is not applicable in vitro.


Normative auditory brainstem response data for hearing threshold in the rabbit.
In an experimental study, we determined the physiological hearing threshold of the rabbit in order to use these data as normative values for further experimental investigations. The aim was to use different acoustic stimuli (click and tone-pip stimuli) with different frequency spectra for air and bone conduction (BC) in order to obtain further information about the optimal form of stimulus when recording auditory evoked potentials in the rabbit. For the investigation, we used 46 female New Zealand rabbits weighing 3.2-4.4 kg and aged 6 months. The equipment used to record brainstem auditory evoked potentials was the Nicolet Viking IV P System (Nicolet Biomedical, Inc.). In accordance with the experimental set-up, the measurements took place under intubation anesthesia, with a total of four repeat measurements performed on each ear at different times. Tone-pip and click stimuli with varying intensities of stimulus, transmitted via air conduction and BC, were applied. The I-IV waves proved the most stable for both stimulus modalities. They were registrable in 98.7% of cases, whereas only 30.2% of the V waves could be recorded. Values averaged from all measurements made throughout the study yielded a potential threshold of 34.8 dB peak equivalent (p.e.) SPL for the click stimulus, 13.8 dB p.e. SPL for the tone-pip stimulus at 8 kHz and 34.2 dB p.e. SPL for the click stimulus transmitted via BC. With regard to latencies, the results indicated a good reproducibility through different stimuli with acceptable standard deviations. The values for physiological hearing threshold obtained here can serve as normative data in subsequent experimental animal studies.


Unique roles for E2F1 in the mouse lens in the absence of functional pRB proteins.
Normal lens fiber cell differentiation requires functional retinoblastoma protein (pRB), because inactivation of this protein results in proliferation and apoptosis in normally postmitotic, differentiating fiber cells. Loss of either E2F1 or -3 can partially rescue the lens phenotype in Rb-deficient mice, implying that these E2Fs may have specific targets in this system. The purpose of this study was to determine what unique role E2F1 may play. Expression of E2F family members and target genes was analyzed in the lenses of nontransgenic, E2F1-null, alphaAE7;E2F1-sufficient; and alphaAE7;E2F1-null mice by in situ hybridization, Northern blot analysis, and RT-PCR. In lenses of E2F1-null mice, there was no change in the expression of E2F-2 to -5 or their target genes, compared with E2F1-sufficient mice. However, in the lens of alphaAE7 mice where pRB proteins are inactivated, expression of E2F2 and -3a was increased. The E2F3a increase, but not that of E2F2, was dependent on E2F1. Expression of E2F target genes was increased with expression of E7 and expression of one of these, p19ARF, was E2F1 dependent. Although in the normal lens there do not appear to be unique roles for E2F1 that cannot be fulfilled by other E2F family members, in the absence of functional pRB proteins, E2F1 is specifically responsible for the increased expression of E2F3a and p19ARF. These findings suggest that E2F1 may be the preferred E2F regulating these target genes in the normal lens.


Expression of human hyaluronan synthases in response to external stimuli.
In the present study we have investigated the expression of mRNAs for hyaluronan synthase isoforms (HAS1, HAS2 and HAS3) in different cells in response to various stimuli. Human mesothelial cells, which synthesize large amounts of hyaluronan, express mRNAs encoding all three HAS isoforms, whereas their transformed counterparts, mesothelioma cells, which produce only minute amounts of hyaluronan, express only HAS3 mRNA. Human lung fibroblasts and the glioma cell line U-118 MG express only the HAS2 and HAS3 genes. The expression of the transcripts was higher in subconfluent than in confluent cultures and was well correlated with the production of hyaluronan by the cells. Stimulation of mesothelial cells with platelet-derived growth factor-BB induced an up-regulation of mRNA for HAS2 to a maximum after 6 h of stimulation; HAS1 and HAS3 genes were only induced slightly. Transforming growth factor-beta1 reduced HAS2 mRNA slightly, and hydrocortisone reduced it strongly, within 6 h of stimulation in mesothelial cell cultures but did not significantly affect the expression of mRNAs for HAS1 and HAS3. Induction of HAS1 and HAS2 protein levels in response to the stimuli above correlated with HAS transcript levels. Thus the expression of the three HAS isoforms is more prominent in growing cells than in resting cells and is differentially regulated by various stimuli suggesting distinct functional roles of the three proteins.


Effect of dietary calcium and phosphorus levels on the utilization of iron, copper, and zinc by adult males.
Iron, copper, and zinc utilization were examined in nine adult males fed a moderate calcium-moderate phosphorus diet (MCaMP), a moderate calcium-high phosphorus diet (MCaHP), and a high calcium-high phosphorus diet (HCaHP) during a 39-day balance study. The moderate and high calcium diets contained 780 mg and 2382 mg calcium daily, respectively. The moderate and high phosphorus diets contained 843 and 2442 mg phosphorus daily, respectively. The calcium supplements were fed as calcium gluconate, while the phosphorus supplements were fed as glycerol phosphate. Subjects lost more iron and copper in their feces and apparently retained less iron and copper when fed the HCaHP diet than when fed the other two diets, but these effects were not statistically significant. Urinary iron and copper excretion were significantly affected by the dietary treatments. Dietary treatments had no effect on subjects' fecal and urinary losses of zinc nor on their apparent retention of zinc. Plasma iron, zinc, copper, and transferrin levels and serum ferritin levels were not affected by the dietary treatments.


Doping graphitic and carbon nanotube structures with boron and nitrogen.
Composite sheets and nanotubes of different morphologies containing carbon, boron, and nitrogen were grown in the electric arc discharge between graphite cathodes and amorphous boron-filled graphite anodes in a nitrogen atmosphere. Concentration profiles derived from electron energy-loss line spectra show that boron and nitrogen are correlated in a one-to-one ratio; core energy-loss fine structures reveal small differences compared to pure hexagonal boron nitride. Boron and carbon are anticorrelated, suggesting the substitution of boron and nitrogen into the carbon network. Results indicate that singlephaase CyBxNx as well as separated domains (nanosize) of boron nitride in carbon networks may exist.


Facing the challenge of a changing system: training child welfare workers in a privatized environment.
The state of Kansas' implementation of a privatized child welfare system is arguably an ambitious shift in child welfare service delivery. In an attempt to drastically improve services to vulnerable families, privatization resulted in intended and unintended consequences for the child welfare workforce. Some of these consequences, including the influx of inexperienced new workers, high worker turnover, and managing relationships with multiple partners, are issues that affect training needs of child welfare professionals. The following paper offers one approach to addressing these needs as well as identifying the challenges involved in training in a privatized environment.


The effect of epistasis on the excess of the additive and nonadditive variances after population bottlenecks.
The effect of population bottlenecks on the components of the genetic variance generated by two neutral independent epistatic loci has been studied theoretically (VA, additive; VD, dominant; VAA, additive x additive; VAD, additive x dominant; VDD; dominant x dominant components of variance). Nonoverdominance and overdominance models were considered, covering all possible types of marginal gene action at the single locus level. The variance components in an infinitely large panmictic population (ancestral components) were compared with their expected values at equilibrium, after t consecutive bottlenecks of equal size N (derived components). Formulae were obtained in terms of allele frequencies and effects at each locus and the corresponding epistatic value. An excess of VA after bottlenecks can be assigned to two sources: (1) the spatiotemporal changes in the marginal average effects of gene substitution alpha(i), which are equal to zero only for additive gene action within and between loci; and (2) the covariance between alpha2(i) and the heterozygosity at the loci involved, which is generated by dominance, with or without epistasis. Numerical examples were analyzed, indicating that an increase in VA after bottlenecks will only occur if its ancestral value is minimal or very small. For the nonoverdominance model with weak reinforcing epistasis, that increase has been detected only for extreme frequencies of the negative allele at one or both loci. With strong epistasis, however, this result can be extended to a broad range of intermediate frequencies. With no epistasis, the same qualitative results were found, indicating that dominance can be considered as the primary cause of an increase in VA following bottlenecks. In parallel, the derived total nonadditive variance exceeded its ancestral value (V(NA) = V(D) + V(AA) + V(AD) + V(DD)) for a range of combinations of allele frequencies covering those for an excess of VA and for very large frequencies of the negative allele at both loci. For the overdominance model, an increase in V(A) and V(NA) was respectively observed for equilibrium (intermediate) frequencies at one or both loci or for extreme frequencies at both loci. For all models, the magnitude of the change of V(A) and V(NA) was inversely related to N and t. At low levels of inbreeding, the between-line variance was not affected by the type of gene action. For the models considered, the results indicate that it is unlikely that the rate of evolution may be accelerated after population bottlenecks, in spite of occasional increments of the derived V(A) over its ancestral value.


Notch signaling modulates hypoxia-induced neuroendocrine differentiation of human prostate cancer cells.
Prostate carcinoma is among the most common causes of cancer-related death in men, representing 15% of all male malignancies in developed countries. Neuroendocrine differentiation (NED) has been associated with tumor progression, poor prognosis, and with the androgen-independent status. Currently, no successful therapy exists for advanced, castration-resistant disease. Because hypoxia has been linked to prostate cancer progression and unfavorable outcome, we sought to determine whether hypoxia would impact the degree of neuroendocrine differentiation of prostate cancer cells in vitro. Exposure of LNCaP cells to low oxygen tension induced a neuroendocrine phenotype, associated with an increased expression of the transcription factor neurogenin3 and neuroendocrine markers, such as neuron-specific enolase, chromogranin A, and β3-tubulin. Moreover, hypoxia triggered a significant decrease of Notch 1 and Notch 2 mRNA and protein expression, with subsequent downregulation of Notch-mediated signaling, as shown by reduced levels of the Notch target genes, Hes1 and Hey1. NED was promoted by attenuation of Hes1 transcription, as cells expressing a dominant-negative form of Hes1 displayed increased levels of neuroendocrine markers under normoxic conditions. Although hypoxia downregulated Notch 1 and Notch 2 mRNA transcription and receptor activation also in the androgen-independent cell lines, PC-3 and Du145, it did not change the extent of NED in these cultures, suggesting that androgen sensitivity may be required for transdifferentiation to occur. Hypoxia induces NED of LNCaP cells in vitro, which seems to be driven by the inhibition of Notch signaling with subsequent downregulation of Hes1 transcription.


Recurring pulmonary hamartomas: cause for concern?
We report the case of a well-controlled female asthmatic who developed 'multiple pulmonary hamartomas' on three separate occasions over a period of 25 years that necessitated surgical resection. To our knowledge, this is the first report of recurrent hamartomas in a single individual necessitating multiple thoracotomies.


[Modified version of dissolving suture material on the basis of glycol cellulose ethers: a histological study].
Investigation of manufacturing of home-produced dissolving suture material was conducted. Influence of the material on tissues in the region of threads location and on the organism as a whole was studied up. The material resolved actively, did not cause pathological changes in surrounding tissues and in general pathological processes in the organism of animals. During 180 days the majority of the suture material quantity have resolved.


Physical Activity in Community-Dwelling Older Adults: Which Real-World Accelerometry Measures Are Robust? A Systematic Review.
Measurement of real-world physical activity (PA) data using accelerometry in older adults is informative and clinically relevant, but not without challenges. This review appraises the reliability and validity of accelerometry-based PA measures of older adults collected in real-world conditions. Eight electronic databases were systematically searched, with 13 manuscripts included. Intraclass correlation coefficient (ICC) for inter-rater reliability were: walking duration (0.94 to 0.95), lying duration (0.98 to 0.99), sitting duration (0.78 to 0.99) and standing duration (0.98 to 0.99). ICCs for relative reliability ranged from 0.24 to 0.82 for step counts and 0.48 to 0.86 for active calories. Absolute reliability ranged from 5864 to 10,832 steps and for active calories from 289 to 597 kcal. ICCs for responsiveness for step count were 0.02 to 0.41, and for active calories 0.07 to 0.93. Criterion validity for step count ranged from 0.83 to 0.98. Percentage of agreement for walking ranged from 63.6% to 94.5%; for lying 35.6% to 100%, sitting 79.2% to 100%, and standing 38.6% to 96.1%. Construct validity between step count and criteria for moderate-to-vigorous PA was rs = 0.68 and 0.72. Inter-rater reliability and criterion validity for walking, lying, sitting and standing duration are established. Criterion validity of step count is also established. Clinicians and researchers may use these measures with a limited degree of confidence. Further work is required to establish these properties and to extend the repertoire of PA measures beyond "volume" counts to include more nuanced outcomes such as intensity of movement and duration of postural transitions.


Cullin Ring Ubiquitin Ligases (CRLs) in Cancer: Responses to Ionizing Radiation (IR) Treatment.
Treatment with ionizing radiation (IR) remains the cornerstone of therapy for multiple cancer types, including disseminated and aggressive diseases in the palliative setting. Radiotherapy efficacy could be improved in combination with drugs that regulate the ubiquitin-proteasome system (UPS), many of which are currently being tested in clinical trials. The UPS operates through the covalent attachment of ATP-activated ubiquitin molecules onto substrates following the transfer of ubiquitin from an E1, to an E2, and then to the substrate via an E3 enzyme. The specificity of ubiquitin ligation is dictated by E3 ligases, which select substrates to be ubiquitylated. Among the E3s, cullin ring ubiquitin ligases (CRLs) represent prototypical multi-subunit E3s, which use the cullin subunit as a central assembling scaffold. CRLs have crucial roles in controlling the cell cycle, hypoxia signaling, reactive oxygen species clearance and DNA repair; pivotal factors regulating the cancer and normal tissue response to IR. Here, we summarize the findings on the involvement of CRLs in the response of cancer cells to IR, and we discuss the therapeutic approaches to target the CRLs which could be exploited in the clinic.


An Interactive Web-Based Sexual Health Literacy Program for Safe Sex Practice for Female Chinese University Students: Multicenter Randomized Controlled Trial.
Sexual health concerns among young adults worldwide help to motivate preventative practices against sexually transmitted infections. To foster better sexual health, sexual health literacy must be enhanced. Little research has been conducted on the impact of gender power dynamics on sexual health, such as sexual coercion, even though the prevalence of sexual coercion remains high in China. This study describes the development and systematic evaluation of a web-based sexual health literacy intervention called "Smart Girlfriend" for female Chinese university students. A multicenter randomized controlled trial was conducted with 781 female university students at 5 universities with dormitories in Hong Kong. Inclusion criteria were used to select unmarried, female, Chinese university students who were ≥18 years old and had not received a sexual health intervention in the past 12 months. Participants were randomly assigned to 2 groups: one group received an interactive web-based sexual health literacy intervention and the other group received a single webpage of online information about condom use. The intervention content was based on the Health Belief Model and the Continuum of Conflict and Control theory. The primary outcome was self-reported consistency of condom use with every partner at 3-month and 6-month follow-up assessments, analyzed using zero/one inflated beta (ZOIB) regression. The secondary outcome was an appraisal of the knowledge, attitudes, norms, and self-efficacy of condom use using the 25-item Multidimensional Condom Attitudes Scale (MCAS). The intention to treat was applied in analyses. Of 1503 individuals that were screened, 781 (52%) were randomized into 2 groups. The retention rates at the 3-month and 6-month follow-ups were 92% and 91%, respectively. Most participants were born locally (536/746, 72%), and 18% (134/746) self-reported as a sexual minority. ZOIB results regarding the consistency of condom use were not significant [model 1: odds ratio (OR) 2.25 with a 95% credible interval (CrI) of 0.84-6.36; model 2: OR 8.03 (95% CrI 0.22-330.31); model 3: OR 1.21 (95% CrI 0.78-1.86)]. Consistency in the intervention group was 5% higher (95% CI -1.90 to 11.63) than the control group at the 3-month follow-up, and 1% higher (95% CI -5.81 to 8·02) at the 6-month follow-up. MCAS scores at the 3-month follow-up were significantly higher in the intervention group (mean 122.51, SD 15.97) than the control group (mean 119.86, SD 15.85; P=.02). An interactive web-based sexual health literacy program did not significantly increase the consistency of condom use compared to a single webpage of condom use information; however, it did temporarily improve knowledge, attitudes, norms, and self-efficacy regarding condom use. Future revisions of this intervention should be personalized and delivered with a proactive approach. ClinicalTrials.gov NCT03695679; https://clinicaltrials.gov/ct2/show/NCT03695679.


Identifying a creak probability threshold for an irregular pitch period detection algorithm.
Irregular pitch periods (IPPs) are associated with grammatically, pragmatically, and clinically significant types of nonmodal phonation, but are challenging to identify. Automatic detection of IPPs is desirable because accurately hand-identifying IPPs is time-consuming and requires training. The authors evaluated an algorithm developed for creaky voice analysis to automatically identify IPPs in recordings of American English conversational speech. To determine a perceptually relevant threshold probability, frame-by-frame creak probabilities were compared to hand labels, yielding a threshold of approximately 0.02. These results indicate a generally good agreement between hand-labeled IPPs and automatic detection, calling for future work investigating effects of linguistic and prosodic context.


Loop-Mediated Isothermal Amplification-Coupled Glass Nanopore Counting Toward Sensitive and Specific Nucleic Acid Testing.
Solid-state nanopores have shown great promise and achieved tremendous success in label-free single-molecule analysis. However, there are three common challenges in solid-state nanopore sensors, including the nanopore size variations from batch to batch that makes the interpretation of the sensing results difficult, the incorporation of sensor specificity, and the impractical analysis time at low analyte concentration due to diffusion-limited mass transport. Here, we demonstrate a novel loop-mediated isothermal amplification (LAMP)-coupled glass nanopore counting strategy that could effectively address these challenges. By using the glass nanopore in the counting mode (versus the sizing mode), the device fabrication challenge is considerably eased since it allows a certain degree of pore size variations and no surface functionalization is needed. The specific molecule replication effectively breaks the diffusion-limited mass transport thanks to the exponential growth of the target molecules. We show the LAMP-coupled glass nanopore counting has the potential to be used in a qualitative test as well as in a quantitative nucleic acid test. This approach lends itself to most amplification strategies as long as the target template is specifically replicated in numbers. The highly sensitive and specific sensing strategy would open a new avenue for solid-state nanopore sensors toward a new form of compact, rapid, low-cost nucleic acid testing at the point of care.


Targeting of hematopoietic progenitor cells with MR contrast agents.
To label human hematopoietic progenitor cells with various magnetic resonance (MR) imaging contrast agents and to obtain 1.5-T MR images of them. Hematopoietic progenitor cells, labeled with ferumoxides, ferumoxtran, magnetic polysaccharide nanoparticles-transferrin, P7228 liposomes, and gadopentetate dimeglumine liposomes underwent MR imaging with T1- and T2-weighted spin-echo and fast field-echo sequences. Data were analyzed by measuring MR signal intensities and R1 and R2* relaxation rates of labeled cells and nonlabeled control cells. Mean quantitative data for the various contrast agent groups were assessed for significant differences compared with control cells by means of the Scheffe test. As a standard of reference, MR imaging data were compared with electron microscopic and spectrometric data. For all contrast agents, intracellular cytoplasm uptake was demonstrated with electron microscopy and was quantified with spectrometry. When compared with nonlabeled control cells, progenitor cells labeled with iron oxides showed significantly (P <.05) increased R2*. Cells labeled with gadopentetate dimeglumine liposomes showed significantly increased R1. Detection thresholds were 5 x 10(5) cells for gadopentetate dimeglumine liposomes and ferumoxtran, 2.5 x 10(5) cells for ferumoxides and P7228 liposomes, and 1 x 10(5) cells for magnetic polysaccharide nanoparticles-transferrin. Hematopoietic progenitor cells can be labeled with MR contrast agents and can be depicted with a standard 1.5-T MR imager.


Who receives a diagnosis of attention-deficit/ hyperactivity disorder in the United States elementary school population?
To investigate factors that are associated with the probability of attention-deficit/hyperactivity disorder (ADHD) diagnosis among U.S. elementary school children, including child, family, school, and policy factors. Logistic regression was used to estimate relative risks associated with independent variables using a nationally representative sample of 9278 children in the 2002 follow-up of the Early Childhood Longitudinal Survey-Kindergarten Cohort. Most children in the sample were in third grade at this point. Previous ADHD diagnoses by professionals were reported by parent respondents. A total of 5.44% of children were reported to have received an ADHD diagnosis. Girls, black children, and Hispanic children were less likely to have the diagnosis even after controlling for other characteristics. Living with one's biological father was negatively associated with ADHD diagnosis. We also found regional variation in diagnosis with the western region of the United States having significantly lower instances of ADHD cases. Higher diagnosis rates were associated with having an older teacher, and lower rates were associated with having a white teacher, relative to a nonwhite teacher. In schools that were subject to stricter state-level performance accountability laws, we found higher odds of ADHD diagnoses, but we found no differences associated with larger class sizes or the presence of state laws that restrict school personnel from discussing ADHD treatment options with parents. ADHD diagnosis is likely to be influenced by a child's social and school environment as well as exogenous child characteristics. Concerns that increased pressures for school performance are associated with increased ADHD diagnoses may be justified.


Free radical scavenging activity of conjugated linoleic acid as single or mixed isomers.
Conjugated linoleic acids (CLAs) are a mixture of positional and geometric isomers of linoleic acid (LA) and believed to have many positive biological activities. The present study was undertaken to assess the antioxidant activity of cis-9, trans-11 and trans-10, cis-12 as single or mixed CLA isomers at two ratios, 1:6 and 1:13 (trans-10, cis-12/cis-9, trans-11). A microplate reader was used to determine the free radical scavenging properties of CLAs against DPPH radical in ethanol. The kinetic reactions of CLA-DPPH(•) showed that all tested CLAs have exerted radical scavenging activities in a dose-dependent manner and observed to immediately react and quench DPPH radicals at all tested levels and no lag phase was noticed in CLA-DPPH(•) reactions. The median inhibitory concentration (IC₅₀) value for cis-9, trans-11 CLA was observed to be more effective than other tested CLA. Total antioxidant capacity (TAC) of all tested CLAs were less effective radical scavengers as compared to vitamin E and butylated hydroxytoluene, although all tested CLAs were quenched a high amount (P < 0.05) of DPPH free radicals. All tested CLAs have the ability to directly react and quench DPPH free radicals in ethanol. Furthermore, trans-10, cis-12 CLA has greater maximal efficacy than other tested CLAs as free radical scavenger, while cis-9, trans-11 CLA is the most potent isomer to directly react and quench free radicals at low concentrations in the system, suggesting that the free radical scavenging activity of CLA isomers may contribute to their diverse biological activities.


Werner syndrome protein. II. Characterization of the integral 3' --> 5' DNA exonuclease.
In addition to its DNA helicase activity, Werner syndrome protein (WRN) also possesses an exonuclease activity (Shen, J.-C., Gray, M. D., Kamath-Loeb, A. S., Fry, M., Oshima, J., and Loeb, L. A. (1998) J. Biol. Chem. 273, 34139-34144). Here we describe the properties of nearly homogeneous WRN exonuclease. WRN exonuclease hydrolyzes a recessed strand in a partial DNA duplex but does not significantly digest single-stranded DNA, blunt-ended duplex, or a protruding strand of a partial duplex. Although DNA is hydrolyzed in the absence of nucleoside triphosphates, nuclease activity is markedly stimulated by ATP, dATP, or CTP. WRN exonuclease digests DNA with a 3' --> 5' directionality to generate 5'-dNMP products, and DNA strands terminating with either a 3'-OH or 3'-PO4 group are hydrolyzed to similar extents. A recessed DNA strand with a single 3'-terminal mismatch is hydrolyzed more efficiently by WRN than one with a complementary nucleotide, but the enzyme fails to hydrolyze a DNA strand terminating with two mismatched bases. WRN exonuclease is distinguished from known mammalian DNA nucleases by its covalent association with a DNA helicase, preference for a recessed DNA strand, stimulation by ATP, ability to equally digest DNA with 3'-OH or 3'-PO4 termini, and its preferential digestion of DNA with a single 3'-terminal mismatch.


[What can be done to prevent violence and abuse of children with disabilities?].
Children with physical, psychological and sensorial disabilities are known to be more vulnerable to violence and maltreatment, or to be at a greater risk of these forms of abuse. Violence and maltreatment of children with disabilities is part of the general problem of child maltreatment, which is still under-researched and little known as a whole; prevention, in particular, has received little attention. Preventing violence and maltreatment against children with disabilities means preventing physical pain, emotional suffering, difficulties in integration and sequelae additional to those already experienced by children due to their disability, in addition to avoiding great expense to the community. This article presents a review of the literature published in the last 10 years on the various primary and secondary prevention strategies that have taken place in developed countries, with the aim of bringing the subject to public attention, especially that of health professionals, and of serving as a basis for the planning of future research lines and prevention strategies.


Analysis of relationships between the availability of resources and the use of health services in Finland. a cross-sectional study.
The relationship between the availability of resources and the use of health services was studied. This study covered the whole population of Finland. The unit of study was a health center area. The method of analysis was multiple regression involving the following variables: supply of health center physicians; supply of hospital physicians (reflecting the private medical sector); supply of sick-ward beds in old-age homes; supply of health center hospital beds; supply of public health nurses; and supply of general hospital beds. Several background variables also were included in the equations. It was concluded that a considerable change in the pattern of health services use was dependent on the number of physicians working in primary care in health centers. An increase in the number of these physicians and in physicians working at the outpatient departments of general hospitals is associated with a considerable increase in the number of visits per population to these facilities, a reduction in the need to hospitalize the population, and a considerable reduction in the rate of private-sector contact outside hospitals. The role of public health nurses appeared to be relatively important in reducing the number of health center hospital days, which probably is because of the home nursing activities of these nurses in health centers. This study demonstrated a significant relationship between the availability of most health services resource variables and the use of various services when other variables known to affect utilization were controlled for.


Regulation of ryanodine receptor opening by lumenal Ca(2+) underlies quantal Ca(2+) release in PC12 cells.
Graded or "quantal" Ca(2+) release from intracellular stores has been observed in various cell types following activation of either ryanodine receptors (RyR) or inositol 1,4,5-trisphosphate receptors (InsP(3)R). The mechanism causing the release of Ca(2+) stores in direct proportion to the strength of stimulation is unresolved. We investigated the properties of quantal Ca(2+) release evoked by activation of RyR in PC12 cells, and in particular whether the sensitivity of RyR to the agonist caffeine was altered by lumenal Ca(2+). Quantal Ca(2+) release was observed in cells stimulated with 1 to 40 mM caffeine, a range of caffeine concentrations giving a >10-fold change in lumenal Ca(2+) content. The Ca(2+) load of the caffeine-sensitive stores was modulated by allowing them to refill for varying times after complete discharge with maximal caffeine, or by depolarizing the cells with K(+) to enhance their normal steady-state loading. The threshold for RyR activation was sensitized approximately 10-fold as the Ca(2+) load increased from a minimal to a maximal loading. In addition, the fraction of Ca(2+) released by low caffeine concentrations increased. Our data suggest that RyR are sensitive to lumenal Ca(2+) over the full range of Ca(2+) loads that can be achieved in an intact PC12 cell, and that changes in RyR sensitivity may be responsible for the termination of Ca(2+) release underlying the quantal effect.


Study of the in vitro-in vivo correlation of Danshensu and protocatechuic aldehyde in a two-step release system.
A two-step release system (TSRS) for the compound Danshen, which has drug-release behavior that is in accordance with the circadian rhythms of cardiovascular disease, was developed by combining an effervescent osmotic pump tablet and a pulsed-released tablet into one hard capsule by our lab. An in vivo study indicated that after oral administration of TSRS, two peaks of the plasma concentration of both Danshensu (DS) and protocatechuic aldehyde (PA) were observed, which suggested that the drug plasma concentration-time curve could meet the requirements for chronotherapy of cardiovascular disease after the bed-time administration of such a device. High performance liquid chromatography using an ultraviolet (UV) detector was used to simultaneously determine the concentrations of DS and PA in plasma. This method was simple, convenient, and appropriate for the quality control of DS and PA. A linear correlation model was established based on the percent absorbant data and percent in vitro dissolution data. Because the drugs were released from the device in an osmotic pressure-dependent manner and absorbed rapidly, a reasonable linear regression relationship was observed between the in vitro and in vivo performances. The current study highlights the potential use of such a device for chronopharmaceutical drug delivery.


Therapeutic effect of nimodipine on experimental brain injury.
To study the therapeutic effect of nimodipine on experimental brain injury. Experimental and control rabbits were subjected to a closed head injury. In one group nimodipine was given intravenously and the effect evaluated by electron microscopy, brain water content, calcium levels, transcranial Doppler, and intracranial pressure monitoring. In rabbits treated with nimodipine the level of neuronal cytosolic free calcium was markedly decreased. There were less cellular damage and less spasm of the middle cerebral artery seen on electron microscopy. No difference regarding intracranial pressure changes between the two groups was noted. Nimodipine has a protective action on brain injury by blocking a series of pathological reactions induced by neuronal calcium overload, and by reducing the spasm of brain vessels and improving cerebral blood flow.


Effects of catecholamines on hepatic and skeletal muscle mitochondrial respiration after prolonged exposure to faecal peritonitis in pigs.
Use of norepinephrine to increase blood pressure in septic animals has been associated with increased efficiency of hepatic mitochondrial respiration. The aim of this study was to evaluate whether the same effect could be reproduced in isolated hepatic mitochondria after prolonged in vivo exposure to faecal peritonitis. Eighteen pigs were randomized to 27 h of faecal peritonitis and to a control condition (n = 9 each group). At the end, hepatic mitochondria were isolated and incubated for one hour with either norepinephrine or placebo, with and without pretreatment with the specific receptor antagonists prazosin and yohimbine. Mitochondrial state 3 and state 4 respiration were measured for respiratory chain complexes I and II, and state 3 for complex IV using high-resolution respirometry, and respiratory control ratios were calculated. Additionally, skeletal muscle mitochondrial respiration was evaluated after incubation with norepinephrine and dobutamine with and without the respective antagonists (atenolol, propranolol and phentolamine for dobutamine). Faecal peritonitis was characterized by decreasing blood pressure and stroke volume, and maintained systemic oxygen consumption. Neither faecal peritonitis nor any of the drugs or drug combinations had measurable effects on hepatic or skeletal muscle mitochondrial respiration. Norepinephrine did not improve the efficiency of complex I- and complex II-dependent isolated hepatic mitochondrial respiration [respiratory control ratio (RCR) complex I: 5.6 ± 5.3 (placebo) vs. 5.4 ± 4.6 (norepinephrine) in controls and 2.7 ± 2.1 (placebo) vs. 2.9 ± 1.5 (norepinephrine) in septic animals; RCR complex II: 3.5 ± 2.0 (placebo) vs. 3.5 ± 1.8 (norepinephrine) in controls; 2.3 ± 1.6 (placebo) vs. 2.2 ± 1.1 (norepinephrine) in septic animals]. Prolonged faecal peritonitis did not affect either hepatic or skeletal muscle mitochondrial respiration. Subsequent incubation of isolated mitochondria with norepinephrine and dobutamine did not significantly influence their respiration.


[The role of culture in psychiatric epidemiology: an examination of research on Latin American mental health].
In psychiatric epidemiology, culture has often been treated in one of two ways: as a confounding variable to be controlled or as an explanation for unexpected findings. In cases where culture is presented as an explanation, there is often little detailed examination of how culture shapes responses to psychiatric interviews. In this paper, I will argue that culture plays a central role in influencing responses to psychiatric interviews in at least three ways: culture affects the way people respond to psychiatric interviews; culture influences the valuation of specific symptom items; and cultural categories are embedded in responses to psychiatric interviews. Through a review of research on the mental health of Latino populations, I will examine the roles of cultural categories in shaping responses to psychiatric epidemiology studies and will outline an alternative research approach to simultaneously exploring psychiatric diagnostic categories and cultural idioms of distress.


A direct gene transfer strategy via brain internal capsule reverses the biochemical defect in Tay-Sachs disease.
Therapy for neurodegenerative lysosomal Tay-Sachs (TS) disease requires active hexosaminidase (Hex) A production in the central nervous system and an efficient therapeutic approach that can act faster than human disease progression. We combined the efficacy of a non-replicating Herpes simplex vector encoding for the Hex A alpha-subunit (HSV-T0alphaHex) and the anatomic structure of the brain internal capsule to distribute the missing enzyme optimally. With this gene transfer strategy, for the first time, we re-established the Hex A activity and totally removed the GM2 ganglioside storage in both injected and controlateral hemispheres, in the cerebellum and spinal cord of TS animal model in the span of one month's treatment. In our studies, no adverse effects were observed due to the viral vector, injection site or gene expression and on the basis of these results, we feel confident that the same approach could be applied to similar diseases involving an enzyme defect.


Intralesional versus intracoronary administration of glycoprotein IIb/IIIa inhibitors during percutaneous coronary intervention in patients with acute coronary syndromes: A meta-analysis of randomized controlled trials.
Glycoprotein IIb/IIIa inhibitors (GPIs) have been regarded as an adjuvant regimen to deal with no-reflow. However, whether intralesional (IL) administration of GPIs improves myocardial reperfusion without increasing bleeding in patients with acute coronary syndrome (ACS) compared with intracoronary (IC) administration has not been well addressed. Our meta-analysis aimed to evaluate the efficacy and safety of IL versus IC administration of GPIs for patients with ACS during percutaneous coronary intervention. We systematically searched Medline, Embase, the Cochrane Central Register of Controlled Trials, and Cambridge Scientific Abstracts from January 2007 to May 2017. Thrombolysis in Myocardial Infarction (TIMI) 3 flow, corrected TIMI frame count (CTFC), and complete ST-segment resolution (>70%) were selected as the primary outcomes. Major adverse cardiac events (MACEs) were the secondary outcome, and major bleeding complications were the safety outcome. Data analysis was conducted using the Review Manager 5.3 software. Six randomized controlled trials were included in our meta-analysis. Compared with IC, IL obtained better results in terms of TIMI grade 3 flow [odds ratio (OR) 2.29; 95% confidence intervals (CIs) 1.31-4.01; P = .004], CTFC [weighted mean difference (WMD) -4.63; 95% CI -8.82 to -0.43; P = .03], and complete ST-segment resolution (OR 1.55; 95% CI 1.12-2.14; P = .008). There was a trend toward decreased MACE in the IL administration groups, which was not of statistical significance (OR 0.63; 95% CI 0.30-1.31; P = .22). No significant difference was found between the two groups in terms of in-hospital major bleeding events (OR 2.52; 95% CI .66 to 9.62; P = .18). IL administration yielded favorable outcomes in terms of myocardial tissue reperfusion as evidenced by the improved TIMI flow grade, CTFC, complete ST-segment resolution, and decreased MACE without increasing in-hospital major bleeding events. The IL administration of GPIs can be recommended as the preferred regimen to guard against no-reflow.