A colI-specified product, synthesized in newly infected recipients, limits the amount of DNA transferred during conjugation of Escherichia coli K-12.
The amount of ColI DNA transferred between mating cells of Escherichia coli K-12 increased about fourfold when rifampin-resistant donors were mated with sensitive recipients in the presence of the drug. Conjugational synthesis of ColI in dnaB recipients, shown primarily to reflect conversion of the transferred DNA into double-stranded material, was also enhanced when the recipients were treated with either rifampin or streptomycin. It is suggested that the amount of ColI transfer is normally limited by the synthesis of one or more proteins in the newly infected recipients. The protein is thought to be plasmid-specified because rifampin also quadrupled transfer to UV-irradiated recipients which were deficient in the transcription of the resident DNA. Successive strands of ColI appear to be transferred discontinuously, because the transferred DNA accumulated in normal and rifampin-treated recipients in the form of circular and linear monomeric units. Although rifampin treatment of recipients also increased transfer of a second Ialpha plasmid, R144drd-3, by about four times, the drug failed to cause a substantial increase of Flac transfer in comparable matings.


Bacterial flavohemoglobin: a molecular tool to probe mammalian nitric oxide biology.
A wide range of mammalian signaling and stress pathways are mediated by nitric oxide (NO), which is synthesized in vivo by the nitric oxide synthase (NOS) family of enzymes. Experimental manipulations of NO are frequently achieved by either inhibition or activation of endogenous NOS or via providing exogenous NO sources. On the contrary, many microbes consume NO via flavohemoglobin (FlavoHb), a highly efficient NO-dioxygenase that protects from nitrosative stress. Here we report a novel resource for studying NO in mammalian cells by heterologously expressing Escherichia coli FlavoHb within a lentiviral delivery system. This technique boosts endogenous cellular consumption of NO, thus providing a simple and efficacious approach to studying mammalian NO biology that can be employed as both a primary experimental and confirmatory tool.


A linguistic signature of psychological distancing in emotion regulation.
Effective emotion regulation is critical for mental health and well-being, rendering insight into underlying mechanisms that facilitate this crucial skill invaluable. We combined principles of cognitive linguistics and basic affective science to test whether shifting components of one's language might foster effective emotion regulation. In particular, we explored bidirectional relations between emotion regulation and linguistic signatures of psychological distancing. In Study 1, we assessed whether people spontaneously distance their language (i.e., shift their word use to be less socially and temporally proximate) when regulating emotions. Participants transcribed their thoughts while either passively viewing or actively regulating their emotional responses to negative images. Regulation increased linguistic markers of social and temporal distance, and participants who showed greater linguistic distancing were more successful regulators. Study 2 reversed this relation and investigated whether distancing one's language spontaneously regulated one's emotions. Participants wrote about negative images either using psychologically "close" or "distant" language in physical, social, and temporal domains. All 3 domains of linguistic distancing spontaneously reduced negative affect. Distancing language also "bled" across domains (e.g., temporal distancing spontaneously produced social distancing). This suggests that distancing one's language in 1 domain (e.g., reducing use of present-tense verbs) produces shifts in deep representations of psychological distance that are measurable across domains (e.g., reduced use of the word "I"). Results extend understanding of language-emotion interactions and reveal novel strategies for reducing negative affect. (PsycINFO Database Record


Live birth after anonymous donation of twice-cryopreserved embryos that had been stored in liquid nitrogen for a cumulative storage time of approximately 13.5 years.
To report a live birth after transfer of anonymously donated, twice-cryopreserved embryos that had been stored in liquid nitrogen for approximately 13.5 years. Case report. A private assisted reproduction center. A 44-year-old recipient of donated cryopreserved embryos. Anonymous donation of cryopreserved blastocysts for procreation. Live birth after thawing and replacement of re-cryopreserved blastocysts. Fourteen pronuclear-stage embryos and four cleavage-stage embryos were cryopreserved during a primary IVF cycle. In two separate cycles, one cycle for the primary patient and a subsequent cycle for the first embryo donor recipient, the 18 embryos were thawed and grown to the blastocyst stage for transfer. Supernumerary blastocysts (n = 5) not replaced at either of these two thaw cycles were re-cryopreserved and subsequently donated to another embryo donor recipient. Five blastocysts survived the thaw and three were transferred, resulting in a live birth. The embryos were cryopreserved for a cumulative storage time of approximately 4,909 days (13.4 years). The longevity (viability) of cryopreserved embryos maintained in liquid nitrogen remains to be determined; cryopreserved embryo donation for procreation should not be overlooked, regardless of the length of time that embryos remain in cryostorage.


Reduction in spontaneous and iatrogenic preterm births in twin pregnancies during COVID-19 lockdown in Melbourne, Australia: a multicenter cohort study.
Melbourne, Australia, recorded one of the longest and most stringent pandemic lockdowns in 2020, which was associated with an increase in preterm stillbirths among singleton pregnancies. Twin pregnancies may be particularly susceptible to the impacts of pandemic disruptions to maternity care due to their higher background risk of adverse perinatal outcomes. Multicenter retrospective cohort study of all twin pregnancies birthing in public maternity hospitals in Melbourne. Multivariable log-binomial regression models were used to compare perinatal outcomes between a pre-pandemic group to women in whom weeks 20+0 to 40+0 of gestation occurred entirely during one of two lockdown-exposure periods: exposure 1 from 22 March 2020 to 21 March 2021 and exposure 2 from 22 March 2021 to 27 March 2022. Total preterm births < 37 weeks were significantly lower in exposure 1 compared with the pre-pandemic period (63.1% vs 68.3%; adjusted risk ratio 0.92 95% CI 0.87-0.98, p = 0.01). This was mainly driven by fewer spontaneous preterm births (18.9% vs 20.3%; adjusted risk ratio 0.95 95% CI 0.90-0.99, p = 0.04). There were also lower rates of preterm birth < 34 weeks (19.9% vs 23.0%, adjusted risk ratio 0.93 95% CI 0.89-0.98 p = 0.01) and total iatrogenic births for fetal compromise (13.4% vs 20.4%; adjusted risk ratio 0.94 95% CI 0.89-0.98, p = 0.01). There were fewer special care nursery admissions (38.5% vs 43.4%; adjusted risk ratio 0.91 95% CI 0.87-0.95, p < 0.001) but no significant changes in stillbirth (1.5% vs 1.6%; adjusted risk ratio 1.00 95% CI 0.99-1.01, p = 0.82). Compared with the pre-pandemic period, there were more preterm births < 28 weeks and neonatal intensive care unit admissions in exposure 2. Melbourne's first lockdown-exposure period was associated with lower preterm births in twins without significant differences in adverse newborn outcomes. Our findings provide insights into the influences on preterm birth and the optimal timing of delivery for twins.


SlowMo, a digital therapy targeting reasoning in paranoia, versus treatment as usual in the treatment of people who fear harm from others: study protocol for a randomised controlled trial.
Paranoia is one of the most common symptoms of schizophrenia-spectrum disorders, and is associated with significant distress and disruption to the person's life. Developing more effective and accessible psychological interventions for paranoia is a clinical priority. Our research team has approached this challenge in two main ways: firstly, by adopting an interventionist causal approach to increase effectiveness and secondly, by incorporating user-centred inclusive design methods to enhance accessibility and usability. Our resultant new digital intervention, SlowMo, intensively targets a reasoning style associated with paranoia, fast thinking, characterised by jumping to conclusions and belief inflexibility. It consists of an easy-to-use, enjoyable and memorable digital interface. An interactive web-based app facilitates delivery of face-to-face meetings which is then synchronised with an innovative mobile app for use in daily life. We aim to test the clinical efficacy of SlowMo over 24 weeks to determine the mechanisms through which it reduces paranoia, and to identify participant characteristics that moderate its effectiveness. In a parallel-group randomised controlled trial, with 1:1 allocation, 360 participants with distressing persecutory beliefs will be independently randomised to receive either the SlowMo intervention added to treatment as usual (TAU) or TAU, using randomly varying permuted blocks, stratified by paranoia severity and site. Research workers will be blind to therapy allocation. The primary outcome is paranoia severity over 24 weeks; our hypothesised mechanism of change is reasoning; moderators include negative symptoms and working memory; and secondary outcomes include wellbeing, quality of life, and service use. The accessibility, usability and acceptability of the digital platform will be assessed. SlowMo has been developed as the first blended digital therapy to target fears of harm from others through an inclusive design approach. In addition to testing its efficacy, this trial will add to our understanding of psychological mechanisms in paranoia. The study will examine the usability and adherence of a novel digital therapy, including an app for self-management, in a large sample of people affected by severe mental health difficulties. ISRCTN registry, ID: ISRCTN32448671 . Registered prospectively on 30 January 2017. Date assigned 2 February 2017.


Computational modeling of 915 MHz microwave ablation: Comparative assessment of temperature-dependent tissue dielectric models.
The objective of this study is to develop a computational model for simulating 915 MHz microwave ablation (MWA), and verify the simulation predictions of transient temperature profiles against experimental measurements. Due to the limited experimental data characterizing temperature-dependent changes of tissue dielectric properties at 915 MHz, we comparatively assess two temperature-dependent approaches of modeling of dielectric properties: model A- piecewise linear temperature dependencies based on existing, but limited, experimental data, and model B- similar to model A, but augmented with linear decrease in electrical conductivity above 95 °C, as guided by our experimental measurements. The finite element method was used to simulate MWA procedures in liver with a clinical 915 MHz ablation applicator. A coupled electromagnetic-thermal solver incorporating temperature-dependent tissue biophysical properties of liver was implemented. Predictions of the transient temperature profiles and ablation zone dimensions for both model A and model B were compared against experimental measurements in ex vivo bovine liver tissue. Broadband dielectric properties of tissue within different regions of the ablation zone were measured and reported at 915 MHz and 2.45 GHz. Model B yielded peak tissue temperatures in closer agreement with experimental measurements, attributed to the inclusion of decrease in electrical conductivity at elevated temperature. The simulated transverse diameters of the ablation zone predicted by both models were greater than experimental measurements, which may be in part due to the lack of a tissue shrinkage model. At both considered power levels, predictions of transverse ablation zone diameters were in closer agreement with measurements for model B (max. discrepancy of 5 mm at 60 W, and 3 mm at 30 W), compared to model A (max. discrepancy of 9 mm at 60 W, and 6 mm at 30 W). Ablation zone lengths with both models were within 2 mm at 30 W, but overestimated by up to 10 mm at 60 W. The inclusion of decreased electrical conductivity above 95 °C, implemented with model B as guided by our experimental measurements, may be a good approach for approximating the dynamic changes that occur during MWA at 915 MHz. Although a step toward more effectively modeling MWA at 915 MHz, further investigation of the transition in dielectric properties with temperature and tissue shrinkage, especially at high temperatures is needed for more accurate simulations.


ANGIOGENES: knowledge database for protein-coding and noncoding RNA genes in endothelial cells.
Increasing evidence indicates the presence of long noncoding RNAs (lncRNAs) is specific to various cell types. Although lncRNAs are speculated to be more numerous than protein-coding genes, the annotations of lncRNAs remain primitive due to the lack of well-structured schemes for their identification and description. Here, we introduce a new knowledge database "ANGIOGENES" (http://angiogenes.uni-frankfurt.de) to allow for in silico screening of protein-coding genes and lncRNAs expressed in various types of endothelial cells, which are present in all tissues. Using the latest annotations of protein-coding genes and lncRNAs, publicly-available RNA-seq data was analyzed to identify transcripts that are expressed in endothelial cells of human, mouse and zebrafish. The analyzed data were incorporated into ANGIOGENES to provide a one-stop-shop for transcriptomics data to facilitate further biological validation. ANGIOGENES is an intuitive and easy-to-use database to allow in silico screening of expressed, enriched and/or specific endothelial transcripts under various conditions. We anticipate that ANGIOGENES serves as a starting point for functional studies to elucidate the roles of protein-coding genes and lncRNAs in angiogenesis.


Influence of the frequency of nerve stimulation on the metabolism of 3H-norepinephrine released from the perfused cat spleen: differences observed during and after the period of stimulation.
The metabolism of 3H-norepinephrine (3H-NE) released by different frequencies of nerve stimulation was studied in the perfused cat spleen after labeling the endogenous stores with (-)-3H-NE. For a wide range of frequencies of stimulation, unmetabolized 3H-NE represented between 50 and 60% of the total increase in outflow of radioactivity elicited by nerve stimulation. The deaminated glycol, 3,4-dihydroxyphenylglycol (3H-DOPEG), was the main metabolite of 3H-NE released by nerve stimulation. When the increase in outflow of radioactivity was analyzed for the samples collected during nerve stimulation, there was a progressive decrease in the fraction of 3H-NE released which was collected as 3H-metabolites as the frequency of stimulation was increased from 0.5 to 5 Hz. For the samples collected in the poststimulation period, there was no frequency dependence in the metabolism of the released transmitter: approximately 75% of the total overflow of radioactivity was accounted for by the 3H-NE metabolites, particularly 3H-DOPEG. The time course of the metabolism of 3H-NE released by nerve stimulation revealed that 3H-DOPEG formation was rather small during stimulation and that it increased sharply in the poststimulation samples. The selective increase in 3H-DOPEG formation in the poststimulation period is compatible with the view that neuronal uptake of the released transmitter might be increased immediately after nerve stimulation. Inhibition of neuronal uptake by cocaine or by phenoxybenzamine prevented 3H-DOPEG formation from 3H-NE released by nerve stimulation. Yet, in the presence of cocaine, the fractional release of total radioactivity per shock was not increased at either 1, 5 or 30 Hz. These results support the view that a large fraction of the 3H-NE released by stimulation which is recaptured by nerve endings is metabolized to 3H-DOPEG rather than stored for subsequent reuse. The extensive conversion to 3H-DOPEG of 3H-NE released by nerve stimulation suggests that there may be a difference between the process of neuronal uptake under resting conditions and that which operates under conditions of nerve stimulation. This difference may be related to the concentration of the transmitter achieved in the synaptic gap in each experimental condition. Under resting conditions and during perfusion with low concentrations of NE, neuronal uptake in the perfused cat spleen is coupled with vesicular storage. On the other hand, when the extracellular concentration of NE is increased as a result of nerve stimulation, neuronal uptake of NE appears to be coupled with presynaptic metabolism through monoamine oxidase and aldehyde reductase.


Aminoglycosides restore full-length type VII collagen by overcoming premature termination codons: therapeutic implications for dystrophic epidermolysis bullosa.
Patients with recessive dystrophic epidermolysis bullosa (RDEB) have severe, incurable skin fragility, blistering, and multiple skin wounds due to mutations in the gene encoding type VII collagen (C7), the major component of anchoring fibrils mediating epidermal-dermal adherence. Nearly 10-25% of RDEB patients carry nonsense mutations leading to premature stop codons (PTCs) that result in truncated C7. In this study, we evaluated the feasibility of using aminoglycosides to suppress PTCs and induce C7 expression in two RDEB keratinocyte cell lines (Q251X/Q251X and R578X/R906) and two primary RDEB fibroblasts (R578X/R578X and R163X/R1683X). Incubation of these cells with aminoglycosides (geneticin, gentamicin, and paromomycin) resulted in the synthesis and secretion of a full-length C7 in a dose-dependent and sustained manner. Importantly, aminoglycoside-induced C7 reversed the abnormal RDEB cell phenotype and incorporated into the dermal-epidermal junction of skin equivalents. We further demonstrated the general utility of aminoglycoside-mediated readthrough in 293 cells transiently transfected with expression vectors encoding 22 different RDEB nonsense mutations. This is the first study demonstrating that aminoglycosides can induce PTC readthrough and restore functional C7 in RDEB caused by nonsense mutations. Therefore, aminoglycosides may have therapeutic potential for RDEB patients and other inherited skin diseases caused by nonsense mutations.


CRISPR/Cas12a-Responsive Hydrogels for Conjugation-Free and Universal Indicator Release in Colorimetric Detection.
Recent advances have demonstrated the significant potential and advantages to repurpose existing point-of-care reactions/devices to realize portable detection of nonoriginal targets, e.g., pathogen genes. However, pursuing this aim usually requires protein indicator-nucleic acid conjugation via a covalent bond, which may bring drawbacks such as high cost, complicated procedure, and annoying component rebuilding. Herein, we developed a conjugation-free, effective, and universal detection platform called CRIs-gel (CRISPR/Cas12a-Responsive Indicators@RCA hydrogels). Various protein indicators are pre-encapsulated into the hydrogels made of effective and high-yield rolling circle amplification (RCA). Upon a targeting sequence binding with its antisense crRNA, CRISPR/Cas12a starts its trans-cleavage activity to crush the hydrogel, which may directly release the indicator for downstream readout. Two proteins, amylase (GA) and human chorionic gonadotropin (hCG), are successfully used as model indicators to trigger the downstream amylum-I2 color change and pregnancy test strip response. After coupling with upstream isothermal nucleic acid amplification, both portable readouts may detect as few as 2 copies/μL genetic sequences of influenza A virus (FluA), human papilloma virus (HPV), SARS-CoV-2, and influenza B virus (FluB). This conjugation-free CRIs-gel platform is thus simple, sensitive, and universal and can provide innovative insights for portable point-of-care testing (POCT) development.


Archaeal communities in boreal forest tree rhizospheres respond to changing soil temperatures.
Temperature has generally great effects on both the activity and composition of microbial communities in different soils. We tested the impact of soil temperature and three different boreal forest tree species on the archaeal populations in the bulk soil, rhizosphere, and mycorrhizosphere. Scots pine, silver birch, and Norway spruce seedlings were grown in forest humus microcosms at three different temperatures, 7-11.5°C (night-day temperature), 12-16°C, and 16-22°C, of which 12-16°C represents the typical mid-summer soil temperature in Finnish forests. RNA and DNA were extracted from indigenous ectomycorrhiza, non-mycorrhizal long roots, and boreal forest humus and tested for the presence of archaea by nested PCR of the archaeal 16S rRNA gene followed by denaturing gradient gel electrophoresis (DGGE) profiling and sequencing. Methanogenic Euryarchaeota belonging to Methanolobus sp. and Methanosaeta sp. were detected on the roots and mycorrhiza. The most commonly detected archaeal 16S rRNA gene sequences belonged to group I.1c Crenarchaeota, which are typically found in boreal and alpine forest soils. Interestingly, also one sequence belonging to group I.1b Crenarchaeota was detected from Scots pine mycorrhiza although sequences of this group are usually found in agricultural and forest soils in temperate areas. Tree- and temperature-related shifts in the archaeal population structure were observed. A clear decrease in crenarchaeotal DGGE band number was seen with increasing temperature, and correspondingly, the number of euryarchaeotal DGGE bands, mostly methanogens, increased. The greatest diversity of archaeal DGGE bands was detected in Scots pine roots and mycorrhizas. No archaea were detected from humus samples from microcosms without tree seedling, indicating that the archaea found in the mycorrhizosphere and root systems were dependent on the plant host. The detection of archaeal 16S rRNA gene sequences from both RNA and DNA extractions show that the archaeal populations were living and that they may have significant contribution to the methane cycle in boreal forest soil, especially when soil temperatures rise.


Sensory perception of the foot and ankle following therapeutic applications of heat and cold.
Many athletes are treated with hot and cold modalities prior to therapeutic exercise, but the effects of these treatments on sensory perception are not clear. The purpose of this study was to examine the effects of hot and cold treatments on sensory perception. We recruited 21 volunteer subjects, who reported for testing on three separate occasions. One of three treatments was applied to the left ankle and foot each day for 20 minutes: cold immersion, hot immersion, or quiet sitting (control). Three variables were measured following treatment: topagnosis, two-point discrimination, and one-legged balance. We assigned treatments and the testing order according to a Greco Latin square. Data were analyzed using a multivariate analysis of variance (MANOVA). No significant differences were detected for the three dependent measures, suggesting that therapeutic applications of heat and cold do not affect sensory perception. These findings indicate that heat and cold applications can be used prior to therapeutic exercise programs without interfering with normal sensory perception as do other analgesic and anesthetic agents. For example, the hypalgesic effect of cold, which is essential to cryokinetics, can be realized without fear of altered sensory perception.


Serum Bupivacaine Concentration After Periarticular Injection With a Mixture of Liposomal Bupivacaine and Bupivacaine HCl During Total Knee Arthroplasty.
A relatively new technique to reduce postoperative pain for total knee arthroplasty is to inject a mixture of 266 mg of liposomal bupivacaine and 125 mg of 0.25% bupivacaine HCl with epinephrine 1:300,000 around the knee joint at the time of surgery. Currently, no publications report serum bupivacaine concentrations over time after periarticular injection of liposomal mixed with free bupivacaine. This information is important to ensure safe serum bupivacaine concentrations are maintained especially when considering supplemental or rescue peripheral nerve blocks. A total of 40 subjects scheduled for primary unilateral total knee arthroplasty with intraoperative periarticular injection of the liposomal bupivacaine and bupivacaine HCl mixture were included. Total serum bupivacaine concentrations were measured after the last injection at selected time points and calculated by gas chromatography. Quantile regression techniques were used to analyze the data over time. This study is registered with ClinicalTrials.gov (ID NCT02626559). Peak serum concentration ranged from 0.17 to 1.2 μg/mL and occurred from 10 minutes to 48 hours. Across all time points, the 48-hour interval had the highest mean concentration of total serum bupivacaine at 0.55 μg/mL (SD, 0.27). Quantile regression showed total serum bupivacaine concentrations increased over the 48 hours measured. None of the participants demonstrated signs or symptoms of local anesthetic toxicity. Total serum concentrations of bupivacaine after periarticular administration of liposomal bupivacaine mixed with bupivacaine HCl remained below the described toxicity threshold (2.5 μg/mL) within the first 48 hours, and no patients demonstrated signs or symptoms of toxicity. However, peak serum concentration time was not achieved within the 48-hour interval. Additional studies are needed to describe the course of serum bupivacaine levels after 48 hours and to ascertain the risk of toxicity when combining this method of periarticular injection with peripheral nerve blocks.


Parastomal hernia incarceration due to migrated intragastric balloon.
The temporary placement of intragastric balloons is a common method to achieve rapid weight loss before planned metabolic surgery. We report the case of a 48-year-old morbidly obese patient. Ten years ago the patient underwent emergency sigmoidectomy with creation of a double-barreled ileostomy for perforated diverticulitis. Over time he developed a giant parastomal hernia. For preoperative weight reduction before planned restoration of intestinal continuity, an intragastric balloon was inserted 3 years ago. The patient was admitted to our emergency department with peritonism and a septic shock. After computed tomography showing small bowel ileus, laparotomy was performed, revealing marked ischemia of incarcerated small and large intestine. Only postoperatively was the intragastric balloon found in the resected small bowel, causing a mechanical ileus with consecutive incarceration of the bowel. We review the literature on complications due to the migration of intragastric balloons. This clinical case gives a fair warning of the possible deleterious outcome of intragastric balloons, especially in hernia patients.


Sleep disorders in Machado-Joseph disease.
This article provides a description on clinical features and pathophysiology of the main sleep disorders observed in Machado-Joseph disease (MJD). Pathological studies have clearly demonstrated that degenerative process in MJD is widespread in the nervous system, and not restricted to the cerebellum. Nonmotor manifestations are frequent and may include pain, cramps, dysautonomia, cognitive deficits, psychiatric manifestations, olfactory deficits, fatigue, nutritional issues, and sleep disorders. Sleep disorders are frequent in MJD, and include restless legs syndrome, rapid eye movement sleep behavior disorder, excessive daytime sleepiness, insomnia, sleep apnea, periodic limb movements during sleep, parasomnia, and others. Pathophysiological mechanisms related to sleep disorders in Machado-Joseph are complex and poorly understood. Considering that sleep complaints are a treatable condition, recognizing sleep disorders in MJD is relevant.


Pitx3 regulates tyrosine hydroxylase expression in the substantia nigra and identifies a subgroup of mesencephalic dopaminergic progenitor neurons during mouse development.
Recent studies of mouse mutant aphakia have implicated the homeobox gene Pitx3 in the survival of substantia nigra dopaminergic neurons, the degeneration of which causes Parkinson's disease. To directly investigate a role for Pitx3 in midbrain DA neuron development, we have analysed a line of Pitx3-null mice that also carry an eGFP reporter under the control of the endogenous Pitx3 promoter. We show that the lack of Pitx3 resulted in a loss of nascent substantia nigra dopaminergic neurons at the beginning of their final differentiation. Pitx3 deficiency also caused a loss of tyrosine hydroxylase (TH) expression specifically in the substantia nigra neurons. Therefore, our study provides the first direct evidence that the aphakia allele of Pitx3 is a hypomorph and that Pitx3 is required for the regulation of TH expression in midbrain dopaminergic neurons as well as the generation and/or maintenance of these cells. Furthermore, using the targeted GFP reporter as a midbrain dopaminergic lineage marker, we have identified previously unrecognised ontogenetically distinct subpopulations of dopaminergic cells within the ventral midbrain based on their temporal and topographical expression of Pitx3 and TH. Such an expression pattern may provide the molecular basis for the specific dependence of substantia nigra DA neurons on Pitx3.


Evaluation of putative anti-cryptosporidial drugs in an in vitro culture system.
Cryptosporidium species are major pathogens severely affecting the health of neonate animals, in particular calves, but also cause life-threatening infection of immunocompromised humans. Currently, only halofuginone is approved for prophylactic and metaphylactic treatment of calves but this drug suffers from its limited safety margins. For treatment of immunodeficient human patients, only nitazoxanide and paramomycin are used but data regarding their efficacy are controversial. Aim of the present study was to test a substituted benzimidazole (BAY-AF76184) and a heterocyclic substituted 1,2,4-triazin (BAY-AB24992), both drugs with known anti-protozoal activity, for their potential ability to interfere with Cryptosporidium parvum development in vitro. Development of C. parvum in HCT-8 cells treated with these compounds was compared to negative controls and parasites treated with paromomycin or halofuginone using real-time PCR targeting the 18S rDNAs of parasites and host cells as template. Potential cytotoxic and anti-proliferative effects of drugs were analysed using a lactate dehydrogenase and a WST-1 assay, respectively. BAY-AF76184 and paromomycin dose-dependently inhibited development of C. parvum with EC50 values of 2.37 μM and 69.5 μM, respectively. Although high concentrations of halofuginone and BAY-AB24992 also significantly inhibited parasite development, effects of lower concentrations were very heterogeneous preventing calculation of EC50 values. Halofuginone and BAY-AF76184 dose-dependently interfered with host cell proliferation (EC50 values of 0.35 μM and 9.07 μM, respectively) and the latter also had direct cytotoxic effects (EC50=48.78 μM). However, drug concentrations required for cytopathic were higher than those for effects against C. parvum. Therefore, both BAY-AB24992 and BAY-AF76184 should be considered for further evaluation, e.g. using in vivo models.


A combined method to quantify the retinal metabolic rate of oxygen using photoacoustic ophthalmoscopy and optical coherence tomography.
Quantitatively determining physiological parameters at a microscopic level in the retina furthers the understanding of the molecular pathways of blinding diseases, such as diabetic retinopathy and glaucoma. An essential parameter, which has yet to be quantified noninvasively, is the retinal oxygen metabolic rate (rMRO2). Quantifying rMRO2 is challenging because two parameters, the blood flow rate and hemoglobin oxygen saturation (sO2), must be measured together. We combined photoacoustic ophthalmoscopy (PAOM) with spectral domain-optical coherence tomography (SD-OCT) to tackle this challenge, in which PAOM measured the sO2 and SD-OCT mapped the blood flow rate. We tested the integrated system on normal wild-type rats, in which the measured rMRO2 was 297.86 ± 70.23 nl/minute. This quantitative method may shed new light on both fundamental research and clinical care in ophthalmology in the future.


Structure and optical properties of several organic-inorganic hybrids containing corner-sharing chains of bismuth iodide octahedra.
Two organic-inorganic bismuth iodides of the form (H3N-R-NH3)BiI5 are reported, each containing long and relatively flexible organic groups, R. The norganic framework in each case consists of distorted BiI6 octahedra sharing cis vertexes to form zigzag chains. Crystals of (H3NC18H24S2NH3)BiI5 were grown from a slowly cooled ethylene glycol/2-butanol solution containing bismuth(III) iodide and AETH.2HI, where AETH = 1,6-bis[5'-(2' '-aminoethyl)-2'-thienyl]hexane. The new compound, (H2AETH)BiI5, adopts an orthorhombic (Aba2) cell with the lattice parameters a = 20.427(3) A, b = 35.078(5) A, c = 8.559(1) A, and Z = 8. The structure consists of corrugated layers of BiI5(2-) chains, with Bi-I bond lengths ranging from 2.942(3) to 3.233(3) A, separated by layers of the organic (H2AETH)(2+) cations. Crystals of the analogous (H3NC12H24NH3)BiI5 compound were also prepared from a concentrated aqueous hydriodic acid solution containing bismuth(III) iodide and the 1,12-dodecanediamine (DDDA) salt, DDDA.2HI. (H2DDDA)BiI5 crystallizes in an orthorhombic (Ibam) cell with a = 17.226(2) A, b = 34.277(4) A, c = 8.654(1) A, and Z = 8. The Bi-I bonds range in length from 2.929(1) to 3.271(1) A. While the inorganic chain structure is nearly identical for the two title compounds, as well as for the previously reported (H3NC6H12NH3)BiI5 [i.e., (H2DAH)BiI5] structure, the packing of the chains is strongly influenced by the choice of organic cation. Optical absorption spectra for thermally ablated thin films of the three organic-inorganic hybrids containing BiI5(2-) chains are reported as a function of temperature (25-290 K). The dominant long-wavelength feature in each case is attributed to an exciton band, which is apparent at room temperature and, despite the similar inorganic chain structure, varies in position from 491 to 541 nm (at 25 K).


Constraint satisfaction adaptive neural network and heuristics combined approaches for generalized job-shop scheduling.
This paper presents a constraint satisfaction adaptive neural network, together with several heuristics, to solve the generalized job-shop scheduling problem, one of NP-complete constraint satisfaction problems. The proposed neural network can be easily constructed and can adaptively adjust its weights of connections and biases of units based on the sequence and resource constraints of the job-shop scheduling problem during its processing. Several heuristics that can be combined with the neural network are also presented. In the combined approaches, the neural network is used to obtain feasible solutions, the heuristic algorithms are used to improve the performance of the neural network and the quality of the obtained solutions. Simulations have shown that the proposed neural network and its combined approaches are efficient with respect to the quality of solutions and the solving speed.


Effects of nonmagnetic interparticle forces on magnetorheological fluids.
Effects of nonmagnetic interparticle forces on the on- and off-state behavior of magnetorheological fluids are investigated experimentally and with particle-level simulations. Suspensions of iron particles in an aliphatic oil are modified by surface-active species. The modifications significantly alter the off-state properties, but have little impact on the field-induced stresses. Simulations show similar behavior. Off-state rheological properties are strongly influenced by van der Waals forces and modifications of the short-range repulsive forces. Field-induced stresses are less sensitive to the nonmagnetic forces.


Bone changes in presellar meningioma.
Earlier recognition of presellar meningiomas may be achieved by looking for intracranial hyperplasia of the sphenoid sinus with additional hyperostosis as well as for the better known signs.


Is poor maternal mortality index in Nigeria a problem of care utilization? A case study of Anambra State.
Maternal mortality in Nigeria is unacceptably high. Some of the reasons may include poor socioeconomic development, weak health care system, low socioeconomic status of women and socio-cultural barriers to care utilization. A cross sectional study was carried out to assess the use of maternal services in Anambra State. A multi stage sampling technique was used to select 800 nursing mothers from the State who were interviewed on the use of maternal services during their last pregnancy. Over 95% of the subjects had formal education. Almost all the subjects (99.7%) attended antenatal clinics during their last pregnancy with 92.3% (646) of them making 4 or more visits before delivery. Over 97% (680) of the deliveries took place in formal health facilities. Obviously, the women in the study population utilize maternal health services. The problem of maternal mortality in the country may not necessarily lie with utilization but with the quality of services.


[ADP ribosylation: knowledge and perspectives].
Polyadenosine diphosphate ribose (polyADPR) and polyADPR-polymerase (polyADPR-P) were discovered in our laboratory in 1966. Then it has been demonstrated that the enzyme and the polymer exist in all eukaryotic cells and catalyze a transfer of ADP-ribose moieties to proteins. Ever increasing evidence favours an involvement of the enzyme through protein ADP-ribosylation in DNA duplication, transcription and repair. Recently the existence of poly or oligo ADP-ribosyl transferases were demonstrated in mitochondria and cytoplasmic particles which carry messenger RNA (mRNP). The nuclear DNA dependent ADPR polymerase produces ADP-ribosylation of a great number of nuclear proteins, resulting in polynucleosome relaxation and modulation of several nuclear enzymes activity. The mRNP ADP-ribosyl transferase may be implicated in derepression of the repressed mRNA. Since 1968 enzymes producing monoADP-ribosylation were discovered in several bacterial toxins and since 1975 in animal tissues. Nicotinamide and its structural analogue: 3-aminobenzamide inhibits ADP-ribosylation and by thus causing inhibition of cell proliferation. Some effects of these inhibitors on cell proliferation are reported and the possible use in tumoral growth as an adjuvant of antimitotics is discussed. Thus ADP-ribosylation appears to be a basic multifunctional event with multiple target sites, some of which are of interest for antitumor therapy.


A genomic perspective on the relationship between the Aquificales and the epsilon-Proteobacteria.
The goal of this study was to determine to what extent the Aquificales are related to the epsilon-Proteobacteria. The genome sequence of several members of this group as well as the genome sequence of Aquifex aeolicus are available. In this study we used information extracted from those whole-genome sequences to gain further insights into the relationships between these organisms, including the fraction of shared putative orthologous protein-encoding genes, dinucleotide relative abundance values and the sequences of the 16S rRNA gene and 20 housekeeping genes. The results of our analyses show that it is not straightforward to come to a consistent picture of the phylogenetic position of the order Aquificales but our data clearly show that there is no particularly close relationship between A. aeolicus and the epsilon-Proteobacteria as (i) they do not share more genes with each other than do other distantly related organisms and (ii) they do not share significant sequence similarity in many macromolecules. In addition, there is considerable evidence that confirms the placement of the Aquificales near the root of the bacterial tree.


Production of oncogene-specific proteins and human T-cell leukemia (lymphotropic) retrovirus (HTLV-I) envelope protein in bacteria and its potential for use in human cancers and seroepidemiological surveys.
The oncogenes coding for the Harvey murine sarcoma virus p21ras protein as well as those coding for myc, myb, and mht products were fused to the amino-terminal portion of the bacteriophage lambda cII gene on the expression vector pJL6. In addition two regions of the gene for the human T-cell leukemia virus subgroup I (HTLV-I) envelope were expressed in our bacterial system. Each of 11 human sera tested that had been shown to contain antibodies to HTLV-I or -II by an enzyme-linked immunosorbent assay recognized the bacterially synthesized envelope proteins. No reaction was detected when 17 control sera were tested. This system will be useful for large-scale seroepidemiological surveys for HTLV-I and related human retroviruses. The other oncogene products expressed in our bacterial vector system also demonstrated specific immunoreactivities. In addition to this feature the bacterial ras protein was seen to bind guanosine diphosphate and was capable of autophosphorylation. Taken together these data suggest that the proteins produced with high efficiency by the bacterial expression system can be immunologically recognized as antigens and can in part perform some of their associated biochemical functions.


A rare cause of a cholestatic jaundice in a North African teenager.
We report an unusual case of intermittent episodes of cholestasis in a young patient. The cholestatic attacks were preceded in each case by an infection and subsequent antibiotic therapies. After ruling out many possible causes of cholestatic hepatitis, the differential diagnoses were a benign recurrent intrahepatic cholestasis or a drug-induced liver injury. We discuss here the diagnostic approach and interpretation of the genetic analysis.


Color vision and the four-color-map problem.
Four different colors are needed to make maps that avoid adjacent countries of the same color. Because the retinal image is two dimensional, like a map, four dimensions of chromatic experience would also be needed to optimally distinguish regions returning spectrally different light to the eye. We therefore suggest that the organization of human color vision according to four-color classes (reds, greens, blues, and yellows) has arisen as a solution to this logical requirement in topology.


Exploring a multi-exposure-pathway approach to assess human health risk associated with groundwater fluoride exposure in the semi-arid region of east India.
Human health risks associated with groundwater fluoride have been assessed using USEPA method in few parts of India, but those assessments were conducted based on a single value for each parameter, which may lead to certain inaccuracy and uncertainties in results. In this study, a higher degree of accuracy in health risk assessment was achieved through Monte Carlo simulations, sensitivity analysis and uncertainty analysis. As fluoride hazards to human health are chronic, a total of 4560 water samples (N = 4560) were collected during consecutive four seasons (2 pre-monsoon and 2 post-monsoon seasons; 1140 samples/season) from the entire Birbhum district, covering all the blocks and geological settings to obtain the spatiotemporal variation of fluoride level. The Empirical Bayesian Kriging geostatistical model was employed to determine fluoride endemic areas. Amongst all blocks, Nalhati-1 had exhibited the highest fluoride level (18.25 mg/L). The study revealed that most of the blocks are vulnerable to groundwater fluoride due to its occurrences in excess level. The average and 95th percentile values of total hazard index indicate that the infants and children populations of the district are more susceptible than the adults and teens. The sensitivity analysis revealed that water ingestion rate and fluoride concentration are the most influential parameters for higher risk of fluoride-related health hazards. Health risks were evaluated through ingestion and dermal exposure routes for infants, children, teens, and adult residents. The exposure was much higher through ingestion than dermal contacts. Ingestion of defluoridated water will reduce the health risks associated with groundwater fluoride exposure.


Genetic overlap between endometriosis and endometrial cancer: evidence from cross-disease genetic correlation and GWAS meta-analyses.
Epidemiological, biological, and molecular data suggest links between endometriosis and endometrial cancer, with recent epidemiological studies providing evidence for an association between a previous diagnosis of endometriosis and risk of endometrial cancer. We used genetic data as an alternative approach to investigate shared biological etiology of these two diseases. Genetic correlation analysis of summary level statistics from genomewide association studies (GWAS) using LD Score regression revealed moderate but significant genetic correlation (rg = 0.23, P = 9.3 × 10-3 ), and SNP effect concordance analysis provided evidence for significant SNP pleiotropy (P = 6.0 × 10-3 ) and concordance in effect direction (P = 2.0 × 10-3 ) between the two diseases. Cross-disease GWAS meta-analysis highlighted 13 distinct loci associated at P ≤ 10-5 with both endometriosis and endometrial cancer, with one locus (SNP rs2475335) located within PTPRD associated at a genomewide significant level (P = 4.9 × 10-8 , OR = 1.11, 95% CI = 1.07-1.15). PTPRD acts in the STAT3 pathway, which has been implicated in both endometriosis and endometrial cancer. This study demonstrates the value of cross-disease genetic analysis to support epidemiological observations and to identify biological pathways of relevance to multiple diseases.


Psychopathy-related differences in selective attention are captured by an early event-related potential.
According to the response modulation model, the poorly regulated behavior of psychopathic individuals reflects a problem reallocating attention to process peripheral information while engaged in goal-directed behavior (Patterson & Newman, 1993). We evaluated this tenet using male prisoners and an early event-related potential component (P140) to index attentional processing. In all task conditions, participants viewed and categorized letter stimuli that could also be used to predict electric shocks. Instructions focused attention either on the threat-relevant dimension of the letters or an alternative, threat-irrelevant dimension. Offenders with high scores on Hare's (2003) Psychopathy Checklist-Revised displayed a larger P140 under alternative versus threat conditions. Beyond demonstrating psychopathy-related differences in early attention, these findings suggest that psychopathic individuals find it easier to ignore threat-related distractors when they are peripheral versus central to their goal-directed behavior.


Social Media and Pathology: Where Are We Now and Why Does it Matter?
Social media has exploded in popularity in recent years. It is a powerful new tool for networking, collaborating, and for the communication and evolution of ideas. It has been increasingly used for business purposes and is now being embraced by physicians including pathologists. Pathology professional organizations and even peer-reviewed pathology journals are now beginning to use social media, as well. There are multiple social media platforms, including Twitter, Facebook, Instagram, LinkedIn, and others. Each platform has different audiences and different ways to share content and interact with other users. This paper discusses the different social media platforms and how they are being used in pathology currently.


[Changes in the neural apparatus of the cerebral arteries in atherosclerosis].
Nervous apparatus of the brain arteries (450-60 microns in diameter) in 55-64-year-old patients with cerebral atherosclerosis was studied. Alterations in the afferent innervation were manifested in the increased sinuosity of fibers, areas of hyper- and hypoimpregnation, appearance of thickenings in the fibers. Changes in the vasomotor innervation consisted of the disturbances of the nerve network structure, an increase of nerve fibers concentration and varicosity in the cholinergic plexuses and their decrease in the adrenergic ones. Restructurings of the nerve apparatus were more pronounced in the arteries of large caliber.


Sorption and desorption of triadimefon by soils and model soil colloids.
Sorption-desorption of the azole fungicide triadimefon [1-(4-chlorophenoxy)-3,3-dimethyl-1-(1H-1,2, 4-triazol-1-yl)-2-butanone] on eight soils and a series of single, binary, and ternary model soil colloids was determined using the batch equilibration technique. Regression analysis between Freundlich sorption coefficients (K(f)) and soil properties suggested that both clay and organic C (OC) were important in triadimefon sorption by soils, with increasing importance of clay for soils with high clay and relatively low OC contents. Triadimefon sorption coefficients on soil were not significantly affected by the concentration of electrolyte or the presence of soluble soil material in solution, but they were highly dependent on the soil:solution ratio due to the nonlinearity of triadimefon sorption on soil. Freundlich sorption isotherms slopes were very similar for all soils (0.75 +/- 0.02). Desorption did not greatly depend on the concentration at which it was determined and showed higher hysteresis for more sorptive soils. Results of triadimefon sorption on model sorbents supported that both humic acid and montmorillonite-type clay constituents contribute to triadimefon retention by soil colloids.


Identification of endogenous control genes for normalisation of real-time quantitative PCR data in colorectal cancer.
Gene expression analysis has many applications in cancer diagnosis, prognosis and therapeutic care. Relative quantification is the most widely adopted approach whereby quantification of gene expression is normalised relative to an endogenously expressed control (EC) gene. Central to the reliable determination of gene expression is the choice of control gene. The purpose of this study was to evaluate a panel of candidate EC genes from which to identify the most stably expressed gene(s) to normalise RQ-PCR data derived from primary colorectal cancer tissue. The expression of thirteen candidate EC genes: B2M, HPRT, GAPDH, ACTB, PPIA, HCRT, SLC25A23, DTX3, APOC4, RTDR1, KRTAP12-3, CHRNB4 and MRPL19 were analysed in a cohort of 64 colorectal tumours and tumour associated normal specimens. CXCL12, FABP1, MUC2 and PDCD4 genes were chosen as target genes against which a comparison of the effect of each EC gene on gene expression could be determined. Data analysis using descriptive statistics, geNorm, NormFinder and qBasePlus indicated significant difference in variances between candidate EC genes. We determined that two genes were required for optimal normalisation and identified B2M and PPIA as the most stably expressed and reliable EC genes. This study identified that the combination of two EC genes (B2M and PPIA) more accurately normalised RQ-PCR data in colorectal tissue. Although these control genes might not be optimal for use in other cancer studies, the approach described herein could serve as a template for the identification of valid ECs in other cancer types.


Gepirone extended-release treatment of anxious depression: evidence from a retrospective subgroup analysis in patients with major depressive disorder.
To evaluate the efficacy and tolerability of gepirone extended-release (ER) tablets in patients with major depressive disorder (MDD) and high ratings of anxiety (anxious depression). This subgroup analysis was derived from an 8-week, double-blind, placebo-controlled study of gepirone ER in patients with MDD. Male and female patients 18 to 69 years of age who met DSM-IV criteria for MDD and had high ratings of anxiety (Hamilton Rating Scale for Depression [HAM-D-17] total score > or = 20 and HAM-D-17 factor I [anxiety/somatization] score > 6) were included in this subgroup analysis. Eligible patients with anxious depression were randomly assigned to receive either placebo or gepirone ER, 20 mg to 80 mg daily. Patient assessments were performed at weeks 1, 2, 3, 4, 6, and 8. Treatment efficacy was evaluated by mean HAM-D-17 total scores and mean changes from baseline in (1) HAM-D-17 total scores, (2) HAM-D-17 factor I (anxiety/somatization) scores, and (3) HAM-D-17 item 12 (anxiety, psychic) scores. All statistical tests were 2-sided and considered statistically significant if the p value was <.05. Between-group comparisons were analyzed using least-squares analysis of variance on the change from baseline at each visit with the last observation carried forward (LOCF). The Cochran-Mantel-Haenszel test adjusting for center was also performed on the percentage of patients in each treatment group at each visit (LOCF) who met the response criterion on the HAM-D-17 (> or = 50% decrease from baseline) or remission criterion (HAM-D-17 total score < or = 7). Gepirone ER-treated patients (N = 58) experienced a statistically significant (p <.05) reduction in mean HAM-D-17 total score at week 3, 6, and 8 compared with placebo-treated patients (N = 75). A statistically significant effect (p <.05) in favor of gepirone ER was observed in mean change from baseline in HAM-D-17 total scores and for HAM-D factor I (anxiety/somatization) scores from week 2 onward. A statistically significant (p < or =.01) effect in favor of gepirone ER was observed in HAM-D-17 item 12 (anxiety, psychic) scores throughout the 8-week trial. There were significantly more patients in the gepirone ER group compared with the placebo group who were HAM-D-17 responders (p <.05) at endpoint and who met the criteria for HAM-D-17 remission at week 3 (p <.05) and weeks 6 and 8 (p <.01). Overall, gepirone ER was well tolerated, with rates of weight gain and sexual dysfunction comparable to placebo. Adverse events were generally mild to moderate. The most commonly reported adverse events were dizziness and nausea. Gepirone ER is an effective and well-tolerated treatment for patients with anxious depression.


Mediastinal pseudocyst with pericardial effusion and dysphagia treated by endoscopic drainage.
Pancreatic pseudocysts located in the mediastinum are rare. Symptomatic mediastinal pseudocysts can present with dysphagia, dyspnea, airway obstruction and/or cardiac tamponade. Generally, the standard approaches are surgery and external drainage. Recently, there have been many reports of successful endoscopic drainage mainly using a transpapillary technique. However, there have only been a handful of reports involving successful transmural drainage of mediastinal pseudocysts. We report a case of a mediastinal pseudocyst developed after a severe blunt trauma. The patient presented with orthopnea and dysphagia. Multidetector computerized scanning of the abdomen and thorax revealed a thin, cystic, low-attenuation mass in the posterior mediastinum associated with compression of the esophagus and significant pericardial effusion. An endoscopic retrograde pancreatogram demonstrated a normal size pancreatic duct with an extravasation of contrast from the tail of the pancreas into the cyst. Ultimately, the cyst was successfully drained trough gastric fundus. Symptomatic mediastinal pseudocysts communicating with the pericardial sac can be successfully drained using a transmural endoscopic approach without the need for surgery or external drainage.


The course of personality pathology.
Durability has traditionally been considered to be a defining feature of personality disorders, but recent studies have challenged this notion. We review the most recent findings on the stability and course of personality pathology. Personality disorders seem to remit more often and faster than previously thought, and their relapse rate is low. However, the recent optimism regarding these disorders is mitigated by the existence of highly heterogeneous trajectories among patients and traits, the identification of certain methodological shortcomings, and the maintenance of psychosocial impairment long after symptomatic relief. The causes of personality improvement are largely unknown, but involve intermingled genetic and environmental effects. Recent follow-up studies of patients with personality pathology are changing orthodox conceptions of their inevitably negative prognosis. The current taxonomies must be reviewed and future research should be integrated with adjacent fields. Treatments need to target the enduring real-life hardships of these patients, apart from their acute symptoms.


In cardiomyocytes, binding of unzipping peptide activates ryanodine receptor 2 and reciprocally inhibits calmodulin binding.
One hypothesis for elevated Ca(2+) leak through cardiac ryanodine receptors (ryanodine receptor 2 [RyR2]) in heart failure is interdomain unzipping that can enhance aberrant channel activation. A peptide (domain peptide corresponding to RyR2 residues 2460-2495 [DPc10]) corresponding to RyR2 central domain residues 2460-2495 recapitulates this arrhythmogenic RyR2 leakiness by unzipping N-terminal and central domains. Calmodulin (CaM) and FK506-binding protein (FKBP12.6) bind to RyR2 and stabilize the closed channel. Little is known about DPc10 binding to the RyR2 and how that may interact with binding (and effects) of CaM and FKBP12.6 to RyR2. To measure, directly in cardiac myocytes, the kinetics and binding affinity of DPc10 to RyR2 and how that affects RyR2 interaction with FKBP12.6 and CaM. We used permeabilized rat ventricular myocytes and fluorescently labeled DPc10, FKBP12.6, and CaM. DPc10 access to its binding site is extremely slow in resting RyR2 but is accelerated by promoting RyR opening or unzipping (by unlabeled DPc10). RyR2-bound CaM (but not FKBP12.6) drastically slowed DPc10 binding. Conversely, DPc10 binding significantly reduced CaM (but not FKBP12.6) binding to the RyR2. Fluorescence resonance energy transfer measurements indicate that DPc10-binding and CaM-binding sites are separate and allow triangulation of the structural DPc10 binding locus on RyR2 vs FKBP12.6-binding and CaM-binding sites. DPc10-RyR2 binding is sterically limited by the resting zipped RyR2 state. CaM binding to RyR2 stabilizes this zipped state, whereas RyR2 activation or prebound DPc10 enhances DPc10 access. DPc10-binding and CaM-binding sites are distinct but are allosterically interacting RyR2 sites. Neither DPc10 nor FKBP12.6 influences RyR2 binding of the other.


A translational fidelity mutation in the universally conserved sarcin/ricin domain of 25S yeast ribosomal RNA.
Recent evidence suggests that ribosomal RNAs have functional roles in translation. We describe here a new ribosomal RNA mutation that causes translational suppression and antibiotic resistance in eukaryotic cells. Using random mutagenesis of the cloned ribosomal RNA gene and in vivo selection, we isolated a C --> U mutation in the universally conserved sarcin/ricin domain in Saccharomyces cerevisiae 25S ribosomal RNA. This mutation changes the putative CG pair, which closes the GAGA tetraloop in the sarcin/ricin domain, into a weaker UG pair without eliminating ribosomal sensitivity to ricin. We show that suppression of several UGA, UAG, and frameshift mutations is evident when a portion of the cellular ribosomal RNA contains the C --> U mutation. Cells that contain essentially all mutant ribosomal RNA grow only 10% slower than the wild-type, but show increased suppression as well as resistance to paramomycin, G418, and hygromycin, and sensitivity to cycloheximide. Our results provide genetic evidence for the participation of the sarcin/ricin loop in maintaining translational accuracy and are discussed in terms of a hypothesis that this ribosomal RNA region normally undergoes a conformational change during translation.


[Bioelectrical activity of the brain, cerebral circulation, and heart rate regulation in the early period of isolated cranio-cerebral injuries].
Combined clinico-physiological examination (electroencephalographic, rheoencephalographic and cardiointervalographic) of 48 victims with early-stage isolated craniocerebral injuries were carried out. From the results of the examination it was concluded that the most marked changes of the brain bioelectric activity in that period were associated with the diminution of the cerebral circulation. The higher frequencies of the EEG oscillations were found to be due to an increase of the sympathoadrenal system activity. The decrease of the sympathoadrenal activity and the diminution of the cerebral circulation intensity observed on the second-fourth day after the injury can be explained as being due to an increase of the permeability of the cerebral vessels and development of brain tissue edema.


[Therapeutic and pharmacological monitoring of individual monotherapy of epilepsy with hexamidine].
The treatment with primidone alone of 53 epileptic patients with generalized tonic-clonic seizures, partial attacks with elementary symptomatology, and partial fits with complex symptomatology removed them completely in 75, 64.2, and 36.9% of the patients, respectively. The number of attacks was decreased as a result by 50% and over in 90, 78.6, and 73.7% of the patients, respectively. The attainment of such high efficacy of the monotherapy (called by the author intensive in contrast to routine or inadequate therapy carried out, as a rule, in standard doses) appeared possible only under the conditions of individualization of the drug doses which ranged within 250-2500 mg/day. The concentration of phenobarbital, the main end metabolite of primidone, in the blood serum of patients, in whom the attacks were completely arrested and who developed no clinical side effects with no toxic action on the internal organs, ranged from 14 to 59.1 mg/l. In partial fits with complex symptomatology, primidone appeared least effective for those types of fits in the treatment of which it was viewed as most adequate.


A history of cigarette smoking is associated with the development of cranial autonomic symptoms with migraine headaches.
To look at the smoking history of migraine patients and to determine if a history of cigarette smoking is associated with the development of cranial autonomic symptoms with migraine headaches. It has recently been noted that a significant number of migraine patients may develop autonomic symptoms during their attacks of headache. Why some headache patients activate the trigeminal autonomic reflex and develop cranial autonomic symptoms while others do not is unknown. Cluster headache occurs more often in patients with a history of cigarette smoking, suggesting a link between tobacco exposure and cluster headache pathogenesis. Could cigarette smoking in some manner lead to activation of the trigeminal-autonomic reflex in headache patients? If cigarette smoking does lower the threshold for activation of the trigeminal autonomic reflex then do migraine patients who have a history of cigarette smoking more often develop cranial autonomic symptoms than migraineurs who have never smoked? Consecutive patients diagnosed with migraine (episodic or chronic) who were seen over a 7-month time period at a newly established headache center were asked about the presence of cranial autonomic symptoms during an attack of head pain. Patients were deemed to have positive autonomic symptoms along with headache if they experienced at least one of the following symptoms: eyelid ptosis or droop, eyelid or orbital swelling, conjunctival injection, lacrimation, or nasal congestion/rhinorrhea. A smoking history was determined for each patient including was the patient a current smoker, past smoker, or had never smoked. Patients were deemed to have a positive history of cigarette smoking if they had smoked continuously during their lifetime for at least at 1 year. A total of 117 migraine patients were included in the analysis (96 female, 21 male). Forty-six patients had a positive smoking history, while 71 patients had no smoking history. Some 70% (32/46) of migraineurs with a positive history of cigarette smoking had cranial autonomic symptoms along with their headaches, while only 42% (30/71) of the nonsmoking patients experienced at least 1 autonomic symptom along with headaches and this was a statistically significant difference (P < .005). In total, 74% of current smokers had autonomic symptoms with their headaches compared with 61% of past smokers and this was not a statistically significant difference. There was a statistically significant difference between the number of current smokers who had autonomic symptoms with their headaches compared with the number of patients who never smoked and had autonomic symptoms (P < .05). Overall, 52% of the studied migraineurs had autonomic symptoms. There was a statistically significant difference between autonomic symptom occurrence in male and female smokers vs male and female nonsmokers. Each subtype of cranial autonomic symptoms was all more frequent in smokers. A history of cigarette smoking appears to be associated with the development of cranial autonomic symptoms with migraine headaches.


Detection of Neospora caninum tachyzoites in cerebrospinal fluid of a dog following prednisone and cyclosporine therapy.
A 9-year-old female spayed Shetland Sheepdog was presented to the Kansas State University Veterinary Medical Teaching Hospital for evaluation following a 3-week history of left rear limb lameness that had progressed to generalized ataxia. Multifocal or diffuse brain lesions were suspected based on physical examination findings. Cerebrospinal fluid (CSF) contained 52 nucleated cells/μL composed of mixed inflammatory cells. Treatment with prednisone and cyclosporine was initiated based on a presumptive diagnosis of granulomatous meningoencephalitis. Thirteen days later the dog was nonambulatory and mentally obtunded. Repeat CSF analysis revealed 298 nucleated cells/μL with 61% eosinophils. Rare protozoal tachyzoites consistent with Neospora caninum, Toxoplasma gondii, or Sarcocystis spp. were found extracellularly and within macrophages and an eosinophil. Despite cessation of prednisone and cyclosporine therapy and provision of supportive care, the dog died 6 days later. Examination of brain tissue sections revealed multifocally extensive, necrotizing, histiocytic, and lymphoplasmacytic meningoencephalitis with numerous protozoal zoites and cysts. Immunohistochemical analysis of brain tissue using a monoclonal antibody specific for N. caninum confirmed the diagnosis of neosporosis. Similar but less severe lesions were noted in the spinal cord, although organisms were not found. This case emphasizes the value of repeated CSF analysis when therapy is ineffective and the importance of excluding infectious causes of meningoencephalitis before commencement of immunosuppressive therapy.


[Pharmacological studies of antispasmodics. III. Antispasmodic activity of 3-(di-2-thienylmethylene)-5-methyl-trans-quino-lizidinium bromide (HRS-902) on smooth muscle organs and its organ selectivity (author's transl)].
Pharmacological activities of HSR-902, a new antispasmodic agent, in gastrointestinal tract, biliary and urinary systems were compared with those of atropine, butylscopolamine bromide, timepidium bromide, prifinium bromide and diphemanil methylsulfate. In isolated smooth muscle organs (stomach, duodenum, ileum, colon, gall bladder and urinary bladder), anti-acetylcholine activities of HSR-902 were the most potent among these agents but its activity in urinary bladder, similar to findings in the cases of atropine and diphemanil methylsulfate, was relatively less potent than that of prifinium bromide, timepidium bromide and butylscopolamine bromide. Regarding the contractions of stomach, jejunum and ileum, which were induced by vagus nerve stimulation or acetylcholine, and on the ileum spontaneous motility, antispasmodic activities of HSR-902 were almost equal to or somewhat more potent than those of atropine, and its activities were more potent than those of prifinium bromide, timepidium bromide, diphemanil methylsulfate and butylscopolamine bromide. On the gall bladder pressure and the counts of perfusion through Oddi's Sphincter, these agents exhibited a similar inhibition and enhancement, respectively. In the case of urinary bladder contractions induced by pelvic nerve stimulation, these agents exhibited a weak inhibition. The inhibitory effect of HSR-902 was relatively less potent than that of other agents except atropine, which had little effect. HSR-902 was similar to atropine in this so-called "atropine-resistance".


Effects of slow repetitive transcranial magnetic stimulation on heart rate variability according to power spectrum analysis.
We examined whether repetitive transcranial magnetic stimulation (rTMS) at a low rate could influence autonomic function, specially heart rate variability (HRV) by power spectrum analysis. We studied 16 healthy male volunteers as a stimulation group and 16 others as a sham group. The stimulation group received magnetic stimulations from a circular coil over Cz at a frequency of 0.2 Hz and an intensity presenting 90% of the motor threshold. Experiments in both groups included four daily sessions; at each, a train of 70 stimuli was delivered over 350 s. HRV of low-frequency power (LF) in a standing position and high-frequency power (HF) in a supine position were measured before and after each session. After stimulation, HF and LF powers were significantly increased. After sham stimulation, the power of HF but not that of LF significantly increased. Neither actual nor sham stimulation produced a long-term effect detectable on day 5. The finding of transiently increased LF power following actual but not sham stimulation suggests that rTMS may activate the sympathetic nervous system.


The role of nitric oxide in saline-induced natriuresis and diuresis in rats.
This study was designed to determine to what extent nitric oxide (NO) mediates the natriuretic and diuretic responses to acute isotonic saline (0.9 gram % NaCl) volume expansion (SVE, 0.5 ml min-1 kg-1). Studies were performed on 49 pentobarbital anesthetized (65 mg/kg) female Sprague-Dawley rats with or without a NO synthase inhibitor, Nomega-nitro-L-arginine (LNA). Group 1 received saline at 27 microliter/min for 1 hr (baseline) and then SVE for 1 hr; Groups 2-4 received LNA at 10, 150, and 200 microgram kg-1 min-1, respectively, for 1 hr followed by LNA + SVE. To determine to what extent inhibition of NOS would reverse an ongoing SVE-induced natriuresis and diuresis, Group 5 was saline-volume-expanded for hours 1 and 2 whereas Group 6 was administered SVE during the first hour and then SVE + 150 microgram kg -1 min-1 LNA during the second hour. SVE caused a significant (P < 0.05) increase in the glomerular filtration rate (GFR) of Group 1 and the LNA-treated rats (Groups 2-4). This SVE-induced increase in the GFR occurred despite the fact that baseline GFR was significantly lower in the two groups of rats that were infused with the highest doses of LNA (Groups 3-4). SVE was also associated with similar increases in urine flow rate, sodium and potassium excretion, and total osmolar excretion in Groups 1-4. On the other hand, mean arterial pressure (MAP) was significantly higher in Group 2 during SVE + LNA and during the baseline as well as during the SVE periods in Groups 3-4; MAP was also significantly elevated in Group 6 during SVE + LNA. Thus, despite the fact that MAP was higher in LNA-treated rats, sodium and urine flow rates were the same as in Group 1 (i.e., there was no evidence of a pressure natriuresis or diuresis in these animals). Along these lines, there was a small but significant positive linear correlation coefficient (r = 0.41, P = 0.05) between sodium excretion values and corresponding MAP values in SVE control rats but not in Groups 3-4 during SVE (r = 0.28, P = 0.26). The current data demonstrate that 1) NO does not mediate SVE-induced hyperfiltration in the rat, 2) NO also does not mediate SVE-induced natriuresis or diuresis, and 3), consistent with other reports, NO appears to mediate pressure natriuresis and diuresis.


Systematic Establishment of Robustness and Standards in Patient-Derived Xenograft Experiments and Analysis.
Patient-derived xenografts (PDX) are tumor-in-mouse models for cancer. PDX collections, such as the NCI PDXNet, are powerful resources for preclinical therapeutic testing. However, variations in experimental and analysis procedures have limited interpretability. To determine the robustness of PDX studies, the PDXNet tested temozolomide drug response for three prevalidated PDX models (sensitive, resistant, and intermediate) across four blinded PDX Development and Trial Centers using independently selected standard operating procedures. Each PDTC was able to correctly identify the sensitive, resistant, and intermediate models, and statistical evaluations were concordant across all groups. We also developed and benchmarked optimized PDX informatics pipelines, and these yielded robust assessments across xenograft biological replicates. These studies show that PDX drug responses and sequence results are reproducible across diverse experimental protocols. In addition, we share the range of experimental procedures that maintained robustness, as well as standardized cloud-based workflows for PDX exome-sequencing and RNA-sequencing analyses and for evaluating growth. SIGNIFICANCE: The PDXNet Consortium shows that PDX drug responses and sequencing results are reproducible across diverse experimental protocols, establishing the potential for multisite preclinical studies to translate into clinical trials.


Cost-effectiveness of misoprostol to control postpartum hemorrhage in low-resource settings.
To test the cost-effectiveness of training traditional birth attendants (TBAs) to recognize postpartum hemorrhage (PPH) and administer a rectal dose of misoprostol in areas with low access to modern delivery facilities. A cost-effectiveness analysis, modeling two hypothetical cohorts of 10,000 women each giving birth with TBAs: one under standard treatment (TBA referral to hospital after blood loss >or=500 ml), and one attended by TBAs trained to recognize PPH and to administer 1000 microg of misoprostol at blood loss >or=500 ml. The misoprostol strategy could prevent 1647 cases of severe PPH (range: 810-2920) and save $115,335 in costs of referral, IV therapy and transfusions (range: $13,991-$1,563,593) per 10,000 births. By preventing severe disease and saving money, it dominates the standard approach. Training TBAs to administer misoprostol to treat PPH has the potential to both save money and improve the health of mothers in low-resource settings.


Genomic organization and deduced amino acid sequence of a putative sodium channel gene in Drosophila.
The deduced amino acid sequence of a Drosophila gene isolated with a vertebrate sodium channel complementary DNA probe revealed an organization virtually identical to the vertebrate sodium channel protein; four homologous domains containing all putative membrane-spanning regions are repeated in tandem with connecting linkers of various sizes. All areas of the protein presumed to be critical for channel function show high evolutionary conservation. These include those proposed to function in voltage-sensitive gating, inactivation, and ion selectivity. All 24 putative gating charges of the vertebrate protein are in identical positions in the Drosophila gene. Ten introns interrupt the coding regions of the four homology units; introns with positions conserved among homology units bracket a region hypothesized to be the selectivity filter for the channel. The Drosophila gene maps to the right arm of the second chromosome in region 60D-E. This position does not coincide with any known mutations that confer behavioral phenotypes, but is close to the seizure locus (60A-B), which has been hypothesized to code for a voltage-sensitive sodium channel.


[Afyon sultandagi earthquake].
Ninety-six percent of Turkey's land is in the earthquake zone. Consequently the results of every earthquake should be presented and discussed. The aim of this study was to evaluate the results of Afyon Sultandagi Earthquake. The records from Governorship of Afyon, local hospitals and Public Work Directorate have been evaluated retrospectively and compared with current literature. An earthquake was detected with the magnitude of 6.0 at 9:11 A.M. at February 3rd, 2002, in the Sultandagi town, which is localized in Afyon city of Turkey. Bolvadin and Cay towns were also affected from the disaster. This earthquake, caused 39 deaths (23 of them were older than 65 year-old; 15 male and 24 female), 325 injured persons; 315 cows and water buffaloes and 3,176 sheep, 14,328 poultry were dead, and 15,032 buildings were damaged. The causes of deaths in human beings were: myocardial infarction in three patients and traumatic cerebral hematoma in one; the others expired from respiratory insufficiency due to dust inhalation because of the traditional architecture of the rural area. Health professionals reached the victims in about four hours, 35 of the injured patients were treated with hospital care and the remaining did not require health support.


Association mapping of genetic risk factors for chronic wasting disease in wild deer.
Chronic wasting disease (CWD) is a fatal transmissible spongiform encephalopathy affecting North American cervids. We assessed the feasibility of association mapping CWD genetic risk factors in wild white-tailed deer (Odocoileus virginianus) and mule deer (Odocoileus hemionus) using a panel of bovine microsatellite markers from three homologous deer linkage groups predicted to contain candidate genes. These markers had a low cross-species amplification rate (27.9%) and showed weak linkage disequilibrium (<1 cM). Markers near the prion protein and the neurofibromin 1 (NF1) genes were suggestively associated with CWD status in white-tailed deer (P = 0.006) and mule deer (P = 0.02), respectively. This is the first time an association between the NF1 region and CWD has been reported.


Treatment of modified Stage II (T1 N1 M0, T2 N1 M0) non-small cell bronchogenic carcinoma. A combined modality approach.
Twenty patients with postsurgical, modified Stage II (T2 N1 M0, T1 N1 M0) non-small cell bronchogenic carcinoma were seen between 1974 and 1981 and were evaluated in a retrospective manner. Fifteen patients had T2 N1 M0 lesions, while 5 patients had T1 N1 M0 disease. Eight patients were treated with surgical resection alone, of whom seven had died, with a median survival of 12.0 months. Four patients received surgical resection and postoperative radiation therapy, of whom two have died, with a median survival not reached at 37 months. Eight patients were treated with surgical resection, radiation therapy, and adjuvant chemotherapy including cyclophosphamide (C), doxorubicin (A), methotrexate (M), and procarbazine (P). Six patients are alive and free of disease, with a median survival not yet reached at 72 months. There is a significant survival advantage for the 12 patients treated with combined modality therapy (surgical resection + radiation therapy; surgical resection + radiation therapy + chemotherapy) compared to the eight patients treated with SR alone (p less than 0.01), and for the eight patients receiving chemotherapy versus the 12 patients who did not (p less than 0.01). In spite of thorough clinical and surgical staging, patients with T1 and T2 primary tumors with N1 disease have a high relapse rate, predominantly in metastatic sites. Adjuvant radiation therapy and chemotherapy appear to benefit these patients with modified Stage II non-small cell bronchogenic carcinoma.


Physical and observational practice afford unique learning opportunities.
In 2 experiments, the authors studied the effectiveness of physical and observational practice on learning and the effect on learning of combining physical practice and observation, as compared with providing physical practice alone. In Experiment 1, retention and transfer performance of 30 university students after physical, observational, or no practice were contrasted. Consistent with findings from other studies, the retention results indicated that observational practice is inferior to physical practice. The transfer data indicated no differences between observation and physical practice groups. In Experiment 2, retention and transfer performance of 30 participants in physical and combined (alternating physical and observational) practice groups were contrasted. The retention results showed no differences between the combined and physical practice groups, but the combined group performed significantly better than the physical practice group on the transfer test. Those findings suggest that a combination of observation and physical practice permits unique opportunities for learning beyond those available via either practice regimen alone.


Comparison of erythropoietin and sildenafil protective role against ischemia/reperfusion injury of the testis in adult rats.
Tissue damage in testicular torsion/detorsion is caused not only by the ischemia, but also by the ischemia/reperfusion injury after detorsion. Erythropoietin and sildenafil are considered to protect against ischemia/reperfusion injury. Here, we studied and compared their actions in testicular torsion/detorsion in adult rats. Twenty-two adult male Wistar Albino rats were divided into four groups. Rats in group A (n = 5) were sham operated. Rats in group B (n = 5), group C (n = 6) and group D (n = 6) underwent torsion of the right testis and detorsion after 90 min. No pharmaceutical intervention was performed in group B. Erythropoietin (1,000 IU/kg) and sildenafil (0.7 mg/kg) were injected intraperitoneally in groups C and D, respectively, after 60 min of torsion. All animals were killed 24 h after detorsion, and their right testis was extracted, placed into 10 % formalin solution and sent for histopathological examination. The histological changes in the testes were scored according to the four-point grading system proposed by Cosentino et al. All rats in group A had normal testicular architecture (grade 1). The untreated group B had a mean grade of 3.81 (range 3.65-4). The treated groups C (mean grade 3.24; range 3.05-3.45) and D (2.69, range 2.4-2.9) presented statistically significant better results (lower grades) compared with the untreated group B. Group D had significantly better results (lower grades) than group C. The intraperitoneal injection of erythropoietin and sildenafil protects against ischemia/reperfusion injury after testicular torsion and detorsion. Sildenafil may have a stronger action than erythropoietin at the doses used in this study.


fhy3-1 retains inductive responses of phytochrome A.
The fhy3 mutation of Arabidopsis impairs phytochrome A (phyA)-mediated inhibition of hypocotyl growth without affecting the levels of phyA measured spectrophotometrically or immunochemically. We investigated whether the fhy3-1 mutation has similar effects on very low fluence responses (VLFR) and high irradiance responses (HIR) of phyA. When exposed to hourly pulses of far-red light, etiolated seedlings of the wild type or of the fhy3-1 mutant showed similar inhibition of hypocotyl growth, unfolding of the cotyledons, anthocyanin synthesis, and greening upon transfer to white light. In the wild type, continuous far-red light was significantly more effective than hourly far-red pulses (at equal total fluence). In the fhy3-1 mutant, hourly pulses were as effective as continuous far-red light, i.e. the failure of reciprocity typical of HIR was not observed. Germination was similarly promoted by continuous or pulsed far-red in wild-type and fhy3-1 seeds. Thus, for hypocotyl growth, cotyledon unfolding, greening, and seed germination, the fhy3-1 mutant retains VLFR but is severely impaired in HIR. These data are consistent with the idea that VLFR and HIR involve divergent signaling pathways of phyA.


Factors affecting assay of myoglobin by complement fixation or immunodiffusion.
Quantitative micro-scale complement-fixation and semi-quantitative gel immunodiffusion assays for myoglobin have been evaluated experimentally and used to measure myoglobin in the sera of more than 1500 patients. We report certain problems, observations, and methodological improvements. Serum caused enhancement of precipitin lines in the immunodiffusion assay. In the complement-fixation assay, serum interfered both by enhancing or inhibiting complement fixation. Suggested modifications in the complement-fixation assay are: (a) routine threefold or greater dilution of serum with buffer and (b) use of serum-based rather than buffer-based calibration materials in preparing standard curves. Three cycles of freeze-thaw of serum or buffer decreased detectable myoglobin by 50-100%. We conclude that certain serum-related factors and freeze-thaw phenomena are important causes of error in immunological assays for myoglobin.


Red blood cell magnesium and chronic fatigue syndrome.
The hypotheses that patients with chronic fatigue syndrome (CFS) have low red blood cell magnesium and that magnesium treatment would improve the wellbeing of such patients were tested in a case-control study and a randomised, double-blind, placebo-controlled trial, respectively. In the case-control study, 20 patients with CFS had lower red cell magnesium concentrations than did 20 healthy control subjects matched for age, sex, and social class (difference 0.1 mmol/l, 95% confidence interval [CI] 0.05 to 0.15). In the clinical trial, 32 patients with CFS were randomly allocated either to intramuscular magnesium sulphate every week for 6 weeks (15 patients) or to placebo (17). Patients treated with magnesium claimed to have improved energy levels, better emotional state, and less pain, as judged by changes in the Nottingham health profile. 12 of the 15 treated patients said that they had benefited from treatment, and in 7 patients energy score improved from the maximum to the minimum. By contrast, 3 of the 17 patients on placebo said that they felt better (difference 62%, 95% CI 35 to 90), and 1 patient had a better energy score. Red cell magnesium returned to normal in all patients on magnesium but in only 1 patient on placebo. The findings show that magnesium may have a role in CFS.


EIF5A2 Promotes Doxorubicin Resistance in Bladder Cancer Cells through the TGF-β Signaling Pathway.
Doxorubicin (DOX) is a commonly used chemotherapeutic agent, but bladder cancer (BC) patients often develop resistance that limits therapeutic efficacy. Recent research has demonstrated a link between medication resistance and the expression of eukaryotic translation initiation factor 5A2 (EIF5A2) in tumors. This study aimed to investigate whether EIF5A2 affects the resistance of BC cells to doxorubicin through the transforming growth factor (TGF)-β signaling pathway. Doxorubicin-resistant cells in BC (T24/DOX and 5637/DOX) were constructed, then cell viability was detected by cell counting kit-8 (CCK-8); EIF5A2 mRNA expression was detected using quantitative real-time PCR (qRT-PCR); cell proliferation was detected using clone formation; apoptosis was detected by flow cytometry; and finally, proteins related to the TGF-β signaling pathway (EIF5A2, TGF-β1, p-small mothers against decapentaplegic 2 (Smad2)/Smad2, p-Smad3/Smad3) were detected using western blot. EIF5A2 was up-regulated in DOX-resistant BC cells, and DOX intervention promoted proliferation and inhibited apoptosis in DOX-resistant BC cells. si-EIF5A2 reversed the above effects. EIF5A2 resulted in DOX resistance by activating the TGF-β pathway, and the TGF-β activator SRI-011381 reversed the inhibitory effect of si-EIF5A2 on DOX resistance. EIF5A2 promotes DOX resistance in BC cells through the TGF-β signaling pathway, and EIF5A2 may be a potential counter-resistance therapeutic strategy in BC chemotherapy.


Quality of life and cost-effectiveness of acupuncture treatment in patients with osteoarthritis pain.
To assess quality of life (QoL), costs, and cost-effectiveness of acupuncture treatment plus routine care versus routine care alone in osteoarthritis patients, a randomised, controlled trial was conducted in 255 general practices in Germany. Four hundred and eighty-nine patients with chronic pain due to osteoarthritis of the knee or hip were included to evaluated QoL and costs at baseline and after 3 months using health insurance funds data and standardized questionnaires. Patients receiving acupuncture had an improved QoL associated with significantly higher costs over the 3 months treatment period compared to routine care alone (mean cost-difference: 469.50 euros [95%CI 135.80-803.19 euros]). This increase in costs was primarily due to the costs of acupuncture. The overall ICER was 17,845 euros per QALY gained. The degree of cost-effectiveness was influenced by gender, with female patients achieving a better cost-effectiveness ratio than men. In conclusion, acupuncture was a cost-effective treatment strategy in patients with chronic osteoarthritis pain.


Allopurinol in the treatment of American cutaneous leishmaniasis.
Pentavalent antimony, the generally accepted treatment for leishmaniasis, is given parenterally, and it is expensive and not readily available in developing countries. An inexpensive, orally administered compound would be a substantial advance in treatment. Previous studies in vitro have shown synergism between allopurinol and pentavalent antimony in tissue-culture systems. We designed this clinical study to determine whether synergism could be demonstrated in patients. We performed a randomized, controlled study of the efficacy of allopurinol plus meglumine antimoniate (Glucantime), as compared with meglumine antimoniate alone, in patients with cutaneous leishmaniasis, who were recruited from a village in southeastern Colombia. In addition, those who declined injections were treated with allopurinol alone, and those who declined any treatment were considered controls. All the patients were followed for one year after the completion of treatment. Lesions that healed completely at three months and remained healed during follow-up were considered to be cured. The cure rate for patients treated with meglumine antimoniate was 36 percent; the addition of allopurinol increased the rate to 74 percent (P less than 0.001). Treatment with allopurinol alone yielded a cure rate of 80 percent (P less than 0.001). There were no cures among the untreated patients. There was no significant difference between the cure rate with allopurinol plus meglumine antimoniate and that with allopurinol alone. No major toxic effects were observed. For the treatment of American cutaneous leishmaniasis, the combination of allopurinol and meglumine antimoniate is significantly more effective than meglumine antimoniate alone, probably because of the efficacy of allopurinol alone, which appears to be as good as the combination.


[Needle aspiration cytology of the breast: current perspective on the role in diagnosis and management].
The aim of this review is to highlight the continuing role of fine needle aspiration cytology (FNAC) in the diagnosis of breast lesions, against a background of its diminishing use in some centres, particularly those involved in breast screening, because of its controversial inadequate rate and suboptimal accuracy. This review explores the current practice and confirms the continuing role of FNAC in the diagnosis and management of breast lesions. The three main areas where FNAC still plays a major role are the following: (a) diagnosis of benign disease in symptomatic palpable lumps as part of triple assessment; (b) staging of breast carcinoma, in particular preoperative axillary lymph node FNAC and intraoperative sentinel node imprints; and (c) diagnosis of metastatic disease at distant sites following treatment for carcinoma. Excision biopsy of the lesion to establish whether it is benign or malignant is not an acceptable mode of diagnosis any more. When triple assessment is concordant, final treatment may be ensued without open biopsy. Triple assessment is a cost effective, easy to perform and time saving approach, however, it can only be used at those institutions where excellent imaging facilities as well as services of a cytopathologist are available. The majority of European countries use similar reporting system for breast FNAC (C1-C5), in keeping with European guidelines for quality assurance in breast cancer screening and diagnosis. A clear reporting system ensures that an unequivocal cytological diagnosis of malignancy is reliable, and in cases where mammography/ultrasonography and clinical examination are in agreement with FNAC, frozen section examination is unnecessary. Suggested thresholds for cytology performance (where therapy is partially based on FNAC) according to the UK NHSBSP are the following: absolute sensitivity (C5 only) >70%, complete sensitivity (C3, C4, C5) >90%, specificity >65%, positive predictive value >99%, false negative <4%, false positive <0.5%, inadequate rate <15%, inadequate rate from cancers <5% and suspicious rate <15%. The issue of optimal sampling to obtain adequate cell material in sufficient quantity is of paramount importance when assessing the accuracy of FNAC. The inadequate rates in FNAC from different sources are lowest when FNAC is performed by a cytopathologist and highest when done by a non-cytopathologist. The multidisciplinary approach is necessary to amplify FNAC quality and to reduce its diagnostic limits. Only when this model of activity is not available, the role of FNAC is less effective and the addition of core biopsy (CB) to FNAC should be considered. CB as an alternative diagnostic modality should be used advisedly, in situations where it is more likely to yield diagnostic information, e.g., in the diagnosis of impalpable masses, microcalcifications or a clinically apparent malignancy where preoperative chemotherapy is planned. CB should not be used as a substitute for poor performance at FNAC. The methods are not mutually exclusive. Where there is access to skilled cytopathologists, FNAC and CB can complement each other and provide a highly accurate, rapid and cost-effective means of patient triage. FNAC has an advantage of being an immediate and excellent method for on-site examination and one-stop diagnosis at breast outpatient clinics. Since the majority of patients attending a breast clinic have benign disease, they benefit from rapid diagnosis and discharge from the clinic. Sentinel node biopsy, now used routinely during the operation for breast carcinoma with the aim of achieving "one-step" surgery can be reduced by one third of patients who ultimately require axillary node dissection if preoperative image guided FNAC of the axilla is used. Positive intraoperative imprint cytology is a reliable tool for proceeding to axillary node dissection, the method having a very high specificity in all published series. FNAC remains the method of choice of diagnosing metastatic disease at extramammary sites. Hormone receptor status, but not HER 2, can be reliably assessed from cytological material. Cells carry a promise of molecular diagnosis and targeted treatment in the future. The future of breast FNAC is bright.


Effects of green tea intake on the development of coronary artery disease.
Green tea, a popular beverage in Japan, contains many polyphenolic antioxidants, which might prevent atherosclerosis. This study was designed to determine whether the consumption of green tea is proportionately associated with a decreased incidence of coronary artery disease (CAD) and the cardiovascular and cerebrovascular prognosis. The study group comprised 203 patients who underwent coronary angiography (109 patients with significant coronary stenosis and 94 patients without). Predictors for CAD were analyzed and the patients' cardiovascular and cerebrovascular events were followed. Green tea consumption was significantly higher in patients without CAD than in those with CAD (5.9+/-0.5 vs 3.5+/-0.3 cups/day; p<0.001). An inverse relationship between the intake of green tea and the incidence of CAD was observed (p<0.001). The green tea intake per day was an independent predictor for CAD based on a multivariate logistic regression analysis (odds ratio: 0.84 and 95% confidence interval: 0.76-0.91). In contrast, the green tea intake was not a predictor of cardiovascular and cerebrovascular events based on the Cox proportional hazard model. Green tea consumption was associated with a lower incidence of CAD in the present study population in Japan. Therefore, the more green tea patients consume, the less likely they are to have CAD.


A highly expressing Tet-inducible cell line recapitulates in situ developmental changes in prestin's Boltzmann characteristics and reveals early maturational events.
Prestin is the motor protein within the lateral membrane of outer hair cells (OHCs), and it is required for mammalian cochlear amplification. Expression of prestin precedes the onset of hearing in mice, and it has been suggested that prestin undergoes a functional maturation within the membrane coincident with the onset of hearing. We have developed a tetracycline-inducible prestin-expressing cell line that we have used to model prestin's functional maturation. We used prestin's voltage-dependent nonlinear charge movement (or nonlinear capacitance) as a test of function and correlated it to biochemical measures of prestin expressed on the cell surface. An initial stage of slow growth in charge density is accompanied by a rapid increase in our estimate of charge carried by an individual motor. A rapid growth in charge density follows and strongly correlates with an increasing ratio between an apparently larger and smaller monomer, suggesting that the latter exerts a dominant-negative effect on function. Finally, there is a gradual depolarizing shift in the voltage of peak capacitance, similar to that observed in developing OHCs. This inducible system offers many opportunities for detailed studies of prestin.