Dual-DMD hyperspectral spatial frequency domain imaging (SFDI) using dispersed broadband illumination with a demonstration of blood stain spectral monitoring.
Spatial frequency domain imaging (SFDI) is a widefield diffuse optical measurement technique capable of generating 2D maps of sub-surface absorption and scattering in biological tissue. We developed a new hyperspectral SFDI instrument capable of collecting images at wavelengths from the visible to the near infrared. The system utilizes a custom-built monochromator with a digital micromirror device (DMD) that can dynamically select illumination wavelength bands from a broadband quartz tungsten halogen lamp, and a second DMD to provide spatially modulated sample illumination. The system is capable of imaging 10 wavelength bands in approximately 25 seconds. The spectral resolution can be varied from 12 to 30 nm by tuning the input slit width and the output DMD column width. We compared the optical property extraction accuracy between the new device and a commercial SFDI system and found an average error of 23% in absorption and 6% in scattering. The system was highly stable, with less than 5% variation in absorption and less than 0.2% variation in scattering across all wavelengths over two hours. The system was used to monitor hyperspectral changes in the optical absorption and reduced scattering spectra of blood exposed to air over 24 hours. This served as a general demonstration of the utility of this system, and points to a potential application for blood stain age estimation. We noted significant changes in both absorption and reduced scattering spectra over multiple discrete stages of aging. To our knowledge, these are the first measurement of changes in scattering of blood stains. This hyperspectral SFDI system holds promise for a multitude of applications in quantitative tissue and diffuse sample imaging.


Oral idarubicin as single-agent treatment of acute nonlymphocytic leukemia in poor-risk patients.
Oral idarubicin was given as single-agent treatment of acute nonlymphocytic leukemia in 18 poor-risk patients. They comprised nine previously untreated elderly patients, age range 69-86, and nine relapsed pretreated patients, age range 41-76. Overall, two patients achieved complete remission (including one with preceding refractory anemia with excess of blasts) and seven achieved partial responses. Dose-limiting toxic effects were diarrhea and sepsis. In this limited study, oral idarubicin at a dose of 20-25 mg/m2/day X 3 was a well-tolerated drug with potent antileukemic effects. The oral formulation deserves more widespread evaluation.


Subthreshold synaptic activation of voltage-gated Ca2+ channels mediates a localized Ca2+ influx into the dendrites of hippocampal pyramidal neurons.
1. Whole cell recordings and high-speed fluorescence imaging were used to investigate the spatial and temporal characteristics of Ca2+ influx during synaptic activity in hippocampal CA1 pyramidal neurons. Brief, subthreshold trains of synaptic potentials elicited by Schaffer collateral stimulation produced transient increases in [Ca2+]i in the apical dendrites near the site of synaptic input. The rises in [Ca2+]i were not due to Ca2+ entry through N-methyl-D-aspartate (NMDA)-activated or non-NMDA-activated glutamate channels, but were reduced by low concentrations of Ni2+. Hyperpolarizing prepulses caused an increase in the synaptically evoked Ca2+ transients, whereas strong hyperpolarization during the train prevented the rise in [Ca2+]i. The data suggest that subthreshold synaptic activity can open low-voltage-activated (T-type) Ca2+ channels and produce a local increase in intradendritic [Ca2+]. Such local increases in [Ca2+]i may be important for modulating the strength of synaptic connections.


Studying the Contribution of Serotonin to Neurodevelopmental Disorders. Can This Fly?
Serotonin is a biogenic amine that acts as neurotransmitter in different brain regions and is involved in complex behaviors, such as aggression or mood regulation. Thus, this amine is found in defined circuits and activates specific receptors in different target regions. Serotonin actions depend on extracellular levels of this amine, which are regulated by its synthetic enzymes and the plasma membrane transporter, SERT. Serotonin acts also as a neurotrophic signal in ontogeny and in the mature brain, controlling cell proliferation, differentiation, neurogenesis, and neural plasticity. Interestingly, early alterations in serotonergic signaling have been linked to a diversity of neurodevelopmental disorders, including autism spectrum disorder (ASD), attention deficit/hyperactivity disorder (ADHD), or mental illnesses like schizophrenia or depression. It has been proposed that given the complex and numerous actions of serotonin, animal models could better serve to study the complexity of serotonin actions, while providing insights on how hindering serotonergic signaling could contribute to brain disorders. In this mini-review, it will be examined what the general properties of serotonin acting as a neurotransmitter in animals are, and furthermore, whether it is possible that Drosophila could be used to study the contribution of this amine to neurodevelopmental and mental disorders.


Does vestibular end-organ function recover after gentamicin-induced trauma in Guinea pigs?
Until 1993 it was commonly accepted that regeneration of vestibular hair cells was not possible in mammals. Two histological studies then showed structural evidence for spontaneous regeneration of vestibular hair cells after gentamicin treatment. There is less evidence for functional recovery going along with this regenerative process; in other words, do regenerated hair cells function adequately? This study aims to address this question, and in general evaluates whether spontaneous functional recovery may occur, in the short or long term, in mammals after ototoxic insult. Guinea pigs were treated with gentamicin for 10 consecutive days at a daily dose of 125 mg/kg body weight. Survival times varied from 1 day to 16 weeks. Vestibular short-latency evoked potentials (VsEPs) to linear acceleration pulses were recorded longitudinally to assess otolith function. After the final functional measurements we performed immunofluorescence histology for hair cell counts. Auditory brainstem responses (ABRs) to click stimuli were recorded to assess cochlear function. As intended, gentamicin treatment resulted in significant loss of utricular hair cells and accompanying declines in VsEPs. Hair cell counts 8 or 16 weeks after treatment did not significantly differ from counts after shorter survival periods. Maximal functional loss was achieved 1-4 weeks after treatment. After this period, only 2 animals showed recovery of VsEP amplitude - all other animals did not reveal signs of regeneration or recovery. In contrast, after initial ABR threshold shifts there was a small but significant recovery. We conclude that spontaneous recovery of otolith function, in contrast to cochlear function, is very limited in guinea pigs. These results support the concept of intratympanic gentamicin treatment where gentamicin is used for chemoablation of the vestibular sensory epithelia.


An efficient site-directed mutagenesis using polymerase chain reaction.
We report here a simple and efficient method for site-directed mutagenesis using polymerase chain reaction (PCR). In constructing a new expression plasmid for the EcoRI restriction gene, we made two point mutations. While one created a new SalI site prior to the SD sequence, the other replaced Glu144 with Lys. A 1.5 kb SalI-PstI fragment isolated from pER101 was used as the template. Two 25 mer oligonucleotide primers containing the desired mutations were synthesized and used to direct PCR amplification with Taq DNA polymerase. About 0.5 microgram of the 0.49 kb fragment was obtained from 0.05 microgram of the 1.5 kb fragment by carrying out polymerase chain reaction for 30 cycles. As calculated theoretically, 99% of the product contained the desired mutations. The product was cloned into pUC19 using SalI and PstI, two of the transformed colonies were randomly chosen for sequence analysis, and both of them were shown to contain the desired mutations. Finally, the amplified fragment was cloned into pER304 to place the EcoRI (Lys144) gene directly under the control of the lambda PL promoter.


Hormone, metabolic peptide, and nutrient levels in the earliest phases of rheumatoid arthritis-contribution of free fatty acids to an increased cardiovascular risk during very early disease.
Rheumatoid arthritis (RA) is a chronic autoimmune disease associated with changes in several hormones and metabolic peptides. Crosstalk between these factors and the immune system may be important for homeostasis during inflammation. Here, we studied the levels of hormones, metabolic peptides, and nutrients in individuals at risk for developing RA (at risk). In total, 18 hormones, metabolic peptides, and nutrients were measured in fasting serum samples from 45 autoantibody-positive individuals at risk, 22 RA patients, and 16 healthy subjects. Triglyceride (TG) levels were also measured in an independent validation cohort of 32 individuals at risk, 20 early arthritis patients, and 20 healthy controls. We found an elevated TG level in individuals at risk and significantly higher TG levels in RA patients compared to healthy controls. These results were confirmed in the validation cohort. Similarly, free fatty acid (FFA) levels showed an increase in individuals at risk and were significantly higher in RA patients compared to healthy controls. In RA patients, FFA levels were positively correlated with disease activity. Pancreatic polypeptide (PP) and norepinephrine levels were highly significantly increased in individuals at risk and RA patients compared to healthy controls. TG and FFA levels are increased in RA patients and positively correlated with disease activity parameters. The results presented here suggest a role for FFAs in the pathogenesis of RA. Furthermore, PP and norepinephrine may be a biomarker that could assist in the identification of individuals at risk.


Results of blind testing a method to detect carriers of the Duchenne muscular dystrophy gene.
We blind-tested a method that in earlier studies had shown increased leucine uptake in muscle fibers of biopsy specimens from three obligate carriers and seven of 11 putative carriers of the gene for Duchenne muscular dystrophy. Here, muscle samples obtained at biopsy in seven obligate carriers and nine control subjects from Brazil were examined in Canada without knowledge of the carrier status or serum enzyme concentrations. Leucine uptake was increased in four controls and within normal range in four carriers, a rate of false-positives and -negatives that underscores the need for blind-testing methods for detecting carriers of this disease.


Pricing for safety and quality in healthcare: A discussion paper.
Increasingly, over the past decade, there has been a global shift in healthcare away from fixed "fee for service" payment mechanisms towards value-based reimbursement models rewarding safety and quality patient outcomes. Curbing the burgeoning costs of healthcare while incentivising higher quality and safer patient care are key drivers of this approach. At face value, this is clearly a worthwhile endeavour. However, there is a lack of conclusive evidence to support the effectiveness of such schemes where they have been introduced internationally. For this reason, Australia has largely been an observer of the shift in payment modalities that are occurring in other countries such as the United States and the United Kingdom. This paper presents an overview of current Australian practice in pricing for safety and quality in Healthcare. Recommendations are provided to help infection control professionals prepare for the upcoming introduction of funding reforms aimed at reducing complications acquired in Australian public hospitals. The implications for infection control professionals are wide-ranging. This will be a period of significant adjustment for the public health system in Australia.


Fabrication and characterization of silicon wire solar cells having ZnO nanorod antireflection coating on Al-doped ZnO seed layer.
In this study, we have fabricated and characterized the silicon [Si] wire solar cells with conformal ZnO nanorod antireflection coating [ARC] grown on a Al-doped ZnO [AZO] seed layer. Vertically aligned Si wire arrays were fabricated by electrochemical etching and, the p-n junction was prepared by spin-on dopant diffusion method. Hydrothermal growth of the ZnO nanorods was followed by AZO film deposition on high aspect ratio Si microwire arrays by atomic layer deposition [ALD]. The introduction of an ALD-deposited AZO film on Si wire arrays not only helps to create the ZnO nanorod arrays, but also has a strong impact on the reduction of surface recombination. The reflectance spectra show that ZnO nanorods were used as an efficient ARC to enhance light absorption by multiple scattering. Also, from the current-voltage results, we found that the combination of the AZO film and ZnO nanorods on Si wire solar cells leads to an increased power conversion efficiency by more than 27% compared to the cells without it.


Surgery and prognostic factors of patients with epidural spinal cord compression caused by hepatocellular carcinoma metastases: retrospective study of 36 patients in a single center.
A retrospective study of 36 patients with metastatic hepatocellular carcinoma (HCC) of the mobile spine was performed by survival analysis. To discuss the factors that may affect outcomes of patients with HCC spinal metastases. HCC is a rare tumor in Western countries. However, HCC is common in Far East (Taiwan, Korea, mainland China), where the hepatitis B virus is epidemic. As the mean survival time of patients with HCC has largely increased in recent years, it is now more common to encounter a patient with epidural spinal cord compression caused by HCC spinal metastases in clinic. The univariate and multivariate analyses of various clinical factors were performed to identify independent variables that could predict prognosis. The survival rate was estimated by the Kaplan-Meier method, and differences were analyzed by the log-rank test. Factors with P values of 0.1 or less were subjected to multivariate analysis for survival rate by multivariate Cox proportional hazards analysis. A total of 36 patients with metastatic HCC of the mobile spine were included in the study. Age (≤45 yr/>45 yr), duration of preoperative symptoms (<6 mo/≥6 mo), preoperative Frankel score (A-C/D-E), Tomita score (5-7/8-10), and bisphosphonate treatment were suggested as the potential prognostic factors through univariate analysis. However, as they were submitted to the multivariate Cox regression model, only Tomita score was found as an independent prognostic factor. Tomita score no more than 7 is a favorable prognostic factor for HCC metastases in the mobile spine. 4.


Acid Production and Proteolytic Activity of Lactobacillus Strains Isolated from Dry Sausages.
The acid-producing capacity and proteolytic activity of 13 strains of Lactobacillus plantarum and 5 strains of Lactobacillus casei isolated from dry sausages was determined at different temperatures and at different NaCl concentrations. Most strains exhibited a maximum acid-producing rate at 30°C. According to the acidification rate at this temperature, strains were divided into three rate groups: fast (I), medium (II) and slow (III), with titratable acidity values above 1.7, between 0.7 and 1.4, and below 0.7, respectively. The decrease in pH ranged between 3.1 and 3.95 according to the group to which the strains belonged. The addition of 3% NaCl produced a marked decrease in the rate of acidification for strains in group II, a slight decrease for those in group I and no effect for those in group III. The proteolytic activity of the strains under study reached a maximum at 40°C, with values between 5.2 and 10 mg% tyrosine released. At 30°C, and in the presence of 3% NaCl, the greatest activity (5.4 mg% tyrosine) was observed in L. plantarum GV 417 and the lowest (3.4 mg% tyrosine) in L. plantarum GV 420. A decrease of approximately 80% in proteolytic activity for all strains was observed in the presence of 5% NaCl.


Magnetic resonance imaging with an optical atomic magnetometer.
We report an approach for the detection of magnetic resonance imaging without superconducting magnets and cryogenics: optical atomic magnetometry. This technique possesses a high sensitivity independent of the strength of the static magnetic field, extending the applicability of magnetic resonance imaging to low magnetic fields and eliminating imaging artifacts associated with high fields. By coupling with a remote-detection scheme, thereby improving the filling factor of the sample, we obtained time-resolved flow images of water with a temporal resolution of 0.1 s and spatial resolutions of 1.6 mm perpendicular to the flow and 4.5 mm along the flow. Potentially inexpensive, compact, and mobile, our technique provides a viable alternative for MRI detection with substantially enhanced sensitivity and time resolution for various situations where traditional MRI is not optimal.


Insular cortex lesions impair the acquisition of conditioned immunosuppression.
Conditioned immunosuppression can be readily obtained in animals by associating a taste with an immunosuppressive drug. On subsequent exposure to the conditioned taste, the animals show an attenuated immune response and also exhibit a conditioned taste aversion. It has been established that insular cortex lesions disrupt the acquisition of conditioned taste aversion. The effect of NMDA-induced lesions in either the insular cortex or the parietal cortex of male Wistar rats was evaluated in the acquisition of conditioned immunosuppression in two experiments. Unlesioned control rats showed the conditioned immunosuppressive response after reexposure to the taste, as indicated by lower hemagglutinating titers to sheep red blood cells in the first experiment and by a decreased IgM production to ovalbumin, measured by ELISA, in the second experiment. Insular cortex-lesioned rats did not show the conditioned immunosuppression in either experiment, while parietal cortex lesions and the sham-lesioned animals presented a clear decrease of hemagglutinating titer and a low IgM production. The insular cortex lesions did not affect the normal immune response, showing normal hemagglutinating titers and IgM production when compared to nonconditioned controls. The immunosuppressive action of cyclophosphamide also remained unaltered. In conclusion, these results show that the insular cortex is essential for the acquisition of conditioned immunosuppression.


[Intravascular lipoma of the superior vena cava].
Lipomas of the major central veins are very rare tumors that have mostly been described in the inferior vena cava. They are often incidentally revealed by computed tomography or magnetic resonance imaging. Histological examination is unusual. We report the case of a lipoma of the superior vena cava discovered on computed tomography performed after a thoracic trauma in a 55-year-old man. The tumor was surgically removed and the fatty nature of the mass was established.


Prediction of short term re-exacerbation in patients with acute exacerbation of chronic obstructive pulmonary disease.
The objective of the study is to develop a scoring system for predicting a 90-day re-exacerbation in hospitalized patients with acute exacerbations of chronic obstructive pulmonary disease (AECOPD). A total of 176 consecutive hospitalized patients with AECOPD were included. The sociodemographic characteristics, status before acute exacerbation (AE), presentations of and treatment for the current AE, and the re-exacerbation in 90 days after discharge from hospital were collected. The re-exacerbation rate in 90 days was 48.9% (86 out of 176). It was associated with the degree of lung function impairment (Global initiative for chronic Obstructive Lung Disease [GOLD] grades), frequency of AE in the previous year, and parameters of the current AE, including pleural effusion, use of accessory respiratory muscles, inhaled long-acting β-2-agonists, inhaled corticosteroids, controlled oxygen therapy, noninvasive mechanical ventilation, and length of hospital stay, but was not associated with body mass index, modified Medical Research Council scale, or chronic obstructive pulmonary disease assessment test. A subgroup of ten variables was selected and developed into the re-exacerbation index scoring system (age grades, GOLD grades, AE times in the previous year, pleural effusion, use of accessory respiratory muscles, noninvasive mechanical ventilation, controlled oxygen therapy, inhaled long-acting β-2-agonists and inhaled corticosteroids, and length of hospital stay). The re-exacerbation index showed good discrimination for re-exacerbation, with a C-statistic of 0.750 (P<0.001). A comprehensive assessment integrating parameters of stable chronic obstructive pulmonary disease, clinical presentations at exacerbation, and treatment showed a strong predictive capacity for short-term outcome in patients with AECOPD. Further studies are required to verify these findings.


Scoliosis vertebral growth plate histomorphometry: Comparisons to controls, growth rates, and compressive stresses.
Scoliosis progression in skeletally immature patients depends on remaining growth. Relationships between vertebral growth plate histomorphometry, growth rates, and mechanical stresses have been reported in several animal studies. Hypertrophic zone heights and chondrocyte heights have been used to assess treatments that aim to modulate growth. The purpose of this study was to determine whether human vertebral physeal hypertrophic zone and cell heights differed between two groups: Severe scoliosis and autopsy controls. Severity was defined at time of surgical planning by curve magnitude and curve stiffness. Physeal samples were obtained from the convex side apex, and from the concave side when feasible. Histologic sections were prepared, and digital images were used to measure hypertrophic zone height, cell height, and cell width. Thirteen spinal deformity patients were included, mean curve magnitude 67° (±23). Etiologies were juvenile and adolescent idiopathic, congenital, neurofibromatosis, neuromuscular, and Marfan syndrome. Five age-matched autopsy specimens without scoliosis served as controls. Results were presented by etiology, then all convex scoliosis specimens were combined and compared to controls. Zone heights for scoliosis, convex side, and controls were 152 µm (±34) and 180 µm (±42) (p = 0.21), cell heights 8.5 µm (±1.1) and 12.8 µm (±1.2) (p < 0.0005), and cell widths 14.9 µm (±1.5) and 15.0 µm (±2.5), respectively. Human values were compared to published animal models and to a quantitative theory of a stress ̶ growth curve. This quantification of vertebral physeal structures in scoliosis may be expected to help assess theories of progression and potential treatments using growth modulation. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:2450-2459, 2018.


Immobilization of Urease onto Modified Egg Shell Membrane through Cross Linking
Immobilization is an approach in industry to improve stability and reusability of urease. The efficiency of this technique depends on the type of membrane and the method of stabilization. The PEI-modified egg shell membrane was used to immobilize urease by absorption and glutaraldehyde cross-linking methods. The membranes were characterized by Fourier-transform infrared spectroscopy (FTIR) and AFM, and Nessler method was applied to measure the kinetic of the immobilized enzymes. Finally, the storage stability (6 °C for 21 days) and reusability (until enzyme activity reached to zero) of the immobilized enzymes were investigated. Based on FTIR, three new peaks were observed in both the absorption- (at 1389.7, 1230.8, and 1074.2 cm-1) and the cross-linking (at 1615-1690, 1392.7, 1450 cm-1) immobilized enzymes. The surface roughness of the native membrane was altered after PEI treatment and enzyme immobilization. The optimal pH of cross-linking immobilized enzymes was shifted to a more neutral pH, while it was alkaline in adsorption-immobilized and free enzymes. The reaction time decreased in all immobilized enzymes (100 min for free enzyme vs. 60 and 30 min after immobilizing by adsorption and cross-linking methods, respectively). The optimal temperature for all enzymes was 70 °C and they had a higher Km and a lower Vmax than free enzyme. The stability and reusability of urease were improved by both methods. Our findings propose these approaches as promising ways to enhance the urease efficiency for its applications in industries and medicines.


The diet of Brycinus nurse (Pisces: Characidae) from Asa reservoir, Ilorin, Nigeria.
From November 1991 to October 1993, 980 specimens of the characid Brycinus nurse were collected from Asa reservoir to examine its diet. The diet was analyzed using the frequency of occurrence, numerical and gravimetric methods. Two hundred and sixty nine (27.45%) of the stomachs examined were empty. The fish was an omnivore feeding extensively on a wide array of plant and animal food items. These consisted of 9 families, 10 genera and 10 species. The most extensively consumed plant food item was aquatic plant parts which occurred in 63.88% of the stomachs, and accounted for 6.06% by number and 12.10% by weight while the ephemeropteran, Povilla adusta was the most dominant animal food item, occurring in 50.92% of the stomachs, and accounting for 11.98% by number and 11.86% by weight. Conversely, the least consumed plant food item was Volvox occurring in 4.49% of the stomachs and accounting for 0.18% by number and 0.35% by weight, while the fish Barbus sp. was the least consumed animal food item occurring in 0.51% of the stomachs, accounting for 0.03% by number and 1.62% by weight. New food items not previously recorded such as a watermite. Aspatharia sinuata and Barbus callipterus were found in the stomach contents. The nonspecific feeding regime of the fish and its ability to utilize different food items effectively was what accounted for the prominence and wide distribution of the fish in the lake.


Krüppel-like factor 8 involved in hypoxia promotes the invasion and metastasis of gastric cancer via epithelial to mesenchymal transition.
Previously, we reported that hypoxia was able to induce invasion and metastasis in gastric cancer and that hypoxia-inducible factor-1 (HIF-1) is a key factor involved in this tumor type. Krüppel-like factor 8 (KLF8) as a transcriptional repressor has been suggested as a promoter of tumor metastasis in breast cancer and an inducer of the epithelial‑mesenchymal transition (EMT). KLF8 is also highly expressed in gastric cancer tissues, contributing to poor prognosis. However, the association between KLF8 and HIF-1 in regulating the progression of human gastric cancer in hypoxia is unclear. In the present study, we found that KLF8 was overexpressed in gastric cancer metastatic tissues and cells. Additionally, KLF8 siRNA significantly inhibited SGC7901 cell invasion and migration compared with SGC7901, SGC7901/Scr-si cells. Hypoxia is thus able to induce KLF8 expression and EMT in SGC7901 cells. However, following the examination of changes in cell morphology and epithelial and mesenchymal markers, it was found that KLF8 siRNA and HIF-1 siRNA strongly reversed EMT in cells undergoing hypoxia. Furthermore, hypoxia-induced KLF8 overexpression was attenuated by HIF-1 siRNA. Experiments using luciferase promoter constructs resulted in a marked increase in the activity of cells exposed to hypoxia and decreased activity in cells co-transfected with HIF-1 siRNA. The chromatin immunoprecipitation assay revealed proximal HRE at -133 is the main HIF-1 binding site in the KLF8 promoter. In conclusion, the results demonstrated that KLF8 is actively enhanced by hypoxia and is a novel HIF-1 target. KLF8 is a novel EMT regulating transcription factor that involved in the progression of gastric cancer. The specific anti-EMT drugs in combination with anti-hypoxia are new promising cancer therapies.


Differential activations of PKC/PKA related to microvasculopathy in diabetic GK rats.
Microvasculopathy is the most serious and predictable threat to the health of diabetic patients, which often results in end-stage renal disease, blindness, and limb amputations. Up to the present, the underlying mechanisms have remained elusive. Here, it was found that the differential activations of PKC/PKA were involved in diabetic microvasculopathy in diabetic GK rats. By real-time PCR, Western blot, immunohistochemistry, and enzyme activity assay, upregulation of PKC was prominent in kidney but was not significant in liver and brain. The expression and activity of PKA were lowered in kidney but comparable in brain and liver during diabetic nephropathy. Furthermore, the generation of reactive oxygen species, production of nitric oxide, and expression of inducible nitric oxide synthase induced by advanced glycation end products were inhibited by PKCβ inhibitor LY-333531 or a PKA agonist in rat glomerular microvascular endothelial cells. Finally, albuminuria was significantly lowered by a PKA agonist and boosted by a PKA antagonist. It suggested that the differential activations of PKC/PKA related to microvasculopathy in diabetes and that activation of PKA may protect the diabetic microvasculature.


Molecular and biochemical analysis of RET/PTC4, a novel oncogenic rearrangement between RET and ELE1 genes, in a post-Chernobyl papillary thyroid cancer.
A post-Chernobyl papillary thyroid cancer, displaying a novel ELE1/RET oncogenic rearrangement with an anomalous fusion transcript, was molecularly characterized. In spite of the presence of a normal breakpoint in exon 5 of the activating ELE1 gene, the sequence of the rearranged genomic DNA showed a previously unreported intra-exonic breakpoint in the RET protooncogene. As a consequence, a cDNA sequence 93 nucleotides larger than the regular one, and with the exon 5 of ELE1 joined to exon 11 instead of exon 12 of RET, is formed. To characterize the product of this new oncogenic ELE1/RET rearrangement, here designated as RET/PTC4, we performed an immunoprecipitation and Western blot analysis on cell extracts from NIH3T3 transfectants. The results showed the presence of two isoforms of the chimeric protein, displaying a constitutive tyrosine phosphorylation. As expected, the molecular weight of this protein was higher than that of RET/ PTC3 protein (p80 and p85, instead of p76 and p81). Previous reports, from our and other laboratories, showed that post-Chernobyl papillary thyroid carcinomas are characterized by a high frequency (about 60%) of RET oncogenic rearrangements (Fugazzola et al., 1995; Klugbauer et al., 1995; Ito et al., 1994). These events predominantly involve ELE1 activating sequence, thus producing RET/PTC3 oncogene (Fugazzola et al., 1995; Klugbauer et al., 1995). Hence, this elevated frequency of RET rearrangements could increase the probability of selecting unusual events as that here described. Alternatively, targeted radiation effects could be responsible for the atypical RET rearrangement producing RET/PTC4 oncogene.


Functional categories and syntactic operations in (Ab)normal language acquisition.
This article argues in favor of the hypothesis that computational complexity determines order of acquisition of functional categories by normal children and patterns of impairment vs. relative preservation of these categories in children with Specific Language Impairment. Complexity is defined in terms of the properties of functional categories of the language the (ab)normal child is acquiring. The results of a study on present/past tense production and comprehension, carried out with children with and without Specific Language Impairment confirm the predictions of the computational complexity hypothesis. It is shown that it is easier to compute kernel functional categories than those that introduce further semantic modifications in the sentence.


Test-and-treat coverage and HIV virulence evolution among men who have sex with men.
HIV set point viral load (SPVL), the viral load established shortly after initial infection, is a proxy for HIV virulence: higher SPVLs lead to higher risk of transmission and faster disease progression. Three models of test-and-treat scenarios, mainly in heterosexual populations, found that increasing treatment coverage selected for more virulent viruses. We modeled virulence evolution in a population of men who have sex with men (MSM) with increasing test-and-treat coverage. We extended a stochastic, dynamic network model (EvoNetHIV). We varied relationship patterns (MSM vs. heterosexual), HIV transmission models (increasing vs. plateauing probability of transmission at very high viral loads), and treatment roll-out (with explicit testing or fixed intervals between infection and treatment). In scenarios most similar to previous models (longer relational durations and the plateauing transmission function), we replicated trends previously found: increasing treatment coverage led to increased virulence (0.12 log10 increase in mean population SPVL between 20% and 100% treatment coverage). In scenarios reflecting MSM behavioral data using the increasing transmission function, increasing treatment coverage selected for viruses with lower virulence (0.16 log10 decrease in mean population SPVL between 20% and 100% treatment coverage). These findings emphasize the impact of sexual network conditions and transmission function details on predicted epidemiological and evolutionary outcomes. Varying these features creates very different evolutionary environments, which in turn lead to opposite effects in mean population SPVL evolution. Our results suggest that, under some realistic conditions, effective test-and-treat strategies may not face the previously reported tradeoff in which increasing coverage leads to evolution of greater virulence. This suggests instead that a virtuous cycle of increasing treatment coverage and diminishing virulence is possible.


[Ventricular dilation during the treatment of subdural hygromas].
Fifty-one cases with subdural hygroma experienced in the past 20 years were retrospectively reviewed. Eight patients showed definite ventricular dilation differing from simple restoration of the ventricles following disappearance of the cavity. The mean age was 72.4 years. Six patients presented with mental change as the initial symptom. On the initial computed tomography (CT), 75% of the cases had bilateral lesions, all were low density, and 88% were crescent shaped. Trepanation performed on six patients yielded watery clear or xanthochromic fluid. Nine to 61 days (mean 4 weeks) after admission, definite ventricular dilation was observed. Cisternography performed in four patients was all abnormal, although cerebrospinal fluid (CSF) pressure was within normal range. Cerebral blood flow images using 123I-iodoamphetamine and single photon emission CT in four patients revealed periventricular low uptake which was disproportionately large compared with the ventricular span on CT. A ventriculoperitoneal shunt was placed in four patients. The final outcome, however, was poor irrespective of treatment. These findings indicate that an impairment of the CSF circulation was not the sole cause of the ventricular dilation. Low CSF pressure and the disproportionately large periventricular low perfusion, compared with the ventricular span on CT scan, suggest a pre-existing periventricular parenchymal damage, which had been subsequently compromised by the presence of subdural mass lesion. Therefore, attention should be paid in aged patients with bilateral low dense, crescent-shaped subdural hygroma, presenting with mental change, for the risk of subsequent ventricular dilation which may affect the functional outcome.


Quantification of Urinary Sex Steroids in the Big Brown Bat (Eptesicus fuscus).
AbstractBats (order Chiroptera) are the second largest group of mammals, diverging ~52.5 million years ago. Many species exhibit an unusual reproductive cycle and extreme longevity without reproductive senescence, yet steroid profiles exist for few bats. Big brown bats (Eptesicus fuscus) are temperate insectivores found throughout North America. They mate promiscuously in fall, store sperm during winter hibernation, and have delayed ovulation and fertilization in spring. Here, we report the first urinary steroid profile in bats by quantifying 17β-estradiol (E2) in captive male and female E. fuscus across their reproductive cycle. Male bats had higher urinary E2 levels than females, and adults had higher levels than yearlings following creatinine adjustment for hydration. In nonpregnant females, several seasonal differences in creatinine-adjusted and unadjusted urinary E2 levels were observed. Urinary E2 was higher in males than females in winter for both conditions and in autumn for creatinine-adjusted levels. We quantified progesterone (P4) in a subset of females. In nonpregnant females, urinary P4 was constant across seasons except for unadjusted levels, which were highest in the summer. In pregnant females, urinary E2 and P4 levels peaked beginning ~20 d before parturition, with both steroids returning to baseline in the following weeks. Knowing how urinary steroid levels fluctuate with age and sex and across the annual season is key to understanding reproductive cycling in bats. Our research furthers the potential for bats as a model for medical reproductive research. Moreover, it complements previous studies on the potential role of steroids in primer pheromonal effects in bats.


Development of a paper-and-pencil semi-adaptive questionnaire for 5 domains of health-related quality of life (PAT-5D-QOL).
To develop a paper-and-pencil semi-adaptive test for 5 domains of health-related quality of life (PAT-5D-QOL) based on item response theory (IRT). The questionnaire uses items from previously developed item banks for 5 domains: (1) walking, (2) handling objects, (3) daily activities, (4) pain or discomfort, and (5) feelings. For each domain, respondents are initially classified into 4 functional levels. Depending on the level, they are instructed to respond to a different set of 5 additional questions. IRT scores for each domain and overall health utility scores are obtained using a simple spreadsheet. The questions were selected using psychometric and conceptual criteria. The format of the questionnaire was developed through focus groups and cognitive interviews. Feasibility was tested in two population surveys. A simulation study was conducted to compare PAT-5D-QOL with a computerized adaptive test (CAT-5D-QOL) and a fixed questionnaire, developed from the same item banks, in terms of accuracy, bias, precision, and ceiling and floor effects. Close to 90 % of the participants in feasibility studies followed the skip instructions properly. In a simulation study, scores on PAT-5D-QOL for all domains tended to be more accurate, more precise, less biased, and less affected by a ceiling effect than scores on a fixed IRT-based questionnaire of the same length. PAT-5D-QOL was slightly inferior to a fully adaptive instrument. PAT-5D-QOL is a novel, semi-adaptive, IRT-based measure of health-related quality of life with a broad range of potential applications.


HIV and Human Coronavirus Coinfections: A Historical Perspective.
Seven human coronaviruses (hCoVs) are known to infect humans. The most recent one, SARS-CoV-2, was isolated and identified in January 2020 from a patient presenting with severe respiratory illness in Wuhan, China. Even though viral coinfections have the potential to influence the resultant disease pattern in the host, very few studies have looked at the disease outcomes in patients infected with both HIV and hCoVs. Groups are now reporting that even though HIV-positive patients can be infected with hCoVs, the likelihood of developing severe CoV-related diseases in these patients is often similar to what is seen in the general population. This review aimed to summarize the current knowledge of coinfections reported for HIV and hCoVs. Moreover, based on the available data, this review aimed to theorize why HIV-positive patients do not frequently develop severe CoV-related diseases.


Ovarian follicular kinetics in response to enucleation and parietal shielding in Rana cyanophlyctis.
Ovarian follicular kinetics, gravimetric changes in the ovary and the oviduct, were studied in the skipper frog, Rana cyanophlyctis, following enucleation (ENX) and parietal shielding (PS), during the breeding reason. PS was to prevent extraoptic photoreception by the pineal via the frontal organ. The gonadosomatic index (GSI) was high in ENX and PS frogs (P < 0.01). The highest GSI occurred in frogs that were both ENX and PS (P < 0.01). Percentage of oviductal weight decreased (P < 0.01) in PS frogs and increased (P < 0.01) in those subjected to both ENX and PS. Oviductal weight of ENX frogs was unchanged. Data on follicular kinetics revealed a decrease (P < 0. 01) in the ENX group in the number of first growth phase (FGP) oocytes and atretic follicles (AF), whereas the number of medium-sized secondary growth phase (MSGP) oocytes and large-sized secondary growth phase (LSGP) oocytes increased (P < 0.01). There was a similar pattern of follicular kinetics in ENX+PS frogs. However, following PS, the number of FGP oocytes and AF increased (P < 0.01) and decreased (P < 0.01) in the MSGP and LSGP oocytes. The lower oviductal weight in PS frogs correlated with reduced SGP oocytes, the major source of estrogen. The importance of lateral eyes and the frontal organs in the transduction of environmental signals to the neuroendocrine-reproductive axis is indicated in this species. That recruitment of FGP to SGP is inhibited only in the PS group suggests that the frontal organ plays an important role in ovarian follicular kinetics.


Effects of regular treatment with combination of salmeterol/fluticasone propionate and salmeterol alone in cough variant asthma.
Cough variant asthma (CVA) is an important cause of chronic cough, and pathophysiological features of the disease appear to be similar to typical asthma. Because CVA is recognized as a precursor of asthma, early intervention with long-term anti-inflammatory agents may be recommended. However, the role of combination therapy with inhaled corticosteroid and β2-agonist in the treatment of CVA has not been elucidated. To evaluate the effectiveness of the combination therapy, we investigated the clinical impact of regular treatment with salmeterol/fliticasone propionate combination (SFC) and inhaled salmeterol (SAL) alone in patients with CVA. The study was a randomized, controlled, parallel-group multi-center trial. Forty-three CVA patients were assigned to SFC (50/100 µg once daily) or SAL (50 µg twice daily) for 12 weeks. Then, these medications were stopped for the next 24 weeks. Main outcome measures were cough symptoms, pulmonary function and airway inflammation. Treatment with each of SFC and SAL significantly decreased cough scores and increased FEV1 and PEF, where the efficacy was more pronounced with SFC than SAL. SFC also decreased sputum eosinophil counts and eosinophil cationic protein contents, whereas SAL had no effect. After discontinuation of the treatment, cough scores increased, pulmonary function and eosinophilic airway inflammation were aggravated and returned to the baseline levels. Maintenance therapy with SFC provides further improvements in cough symptoms, pulmonary function and airway inflammation, and discontinuation of the therapy causes worsening of the disease, indicating that stopping or interrupting anti-inflammatory therapy may not be advisable in patients with CVA.


Typing of Histoplasma capsulatum isolates based on nucleotide sequence variation in the internal transcribed spacer regions of rRNA genes.
The nucleotide sequences of internal transcribed spacer (ITS) regions of rRNA genes of 24 isolates of Histoplasma capsulatum were examined. The results indicate that the sequences of ITS regions in different isolates are not identical. Sequence variations were found at 20 positions in the 496 bp that were sequenced. Ten different sequence patterns, designated types A through H, were observed when the sequences from the 24 isolates were aligned. Twelve isolates from Indianapolis were classified into four different types. Two isolates from New York belonged to type G. Three isolates from different cities were type F. The remaining six isolates were of different types.


Ketone bodies in epilepsy.
Seizures that are resistant to standard medications remain a major clinical problem. One underutilized option for patients with medication-resistant seizures is the high-fat, low-carbohydrate ketogenic diet. The diet received its name based on the observation that patients consuming this diet produce ketone bodies (e.g., acetoacetate, β-hydroxybutyrate, and acetone). Although the exact mechanisms of the diet are unknown, ketone bodies have been hypothesized to contribute to the anticonvulsant and antiepileptic effects. In this review, anticonvulsant properties of ketone bodies and the ketogenic diet are discussed (including GABAergic and glutamatergic effects). Because of the importance of ketone body metabolism in the early stages of life, the effects of ketone bodies on developing neurons in vitro also are discussed. Understanding how ketone bodies exert their effects will help optimize their use in treating epilepsy and other neurological disorders.


Does carbon monoxide treatment alter cytokine levels after endotoxin infusion in pigs? A randomized controlled study.
Carbon monoxide (CO) has recently been suggested to have anti-inflammatory properties, but data seem to be contradictory and species-specific. Thus, in studies on macrophages and mice, pretreatment with CO attenuated the inflammatory response after endotoxin exposure. On the other hand, human studies showed no effect of CO on the inflammatory response. Anti-inflammatory efficacy of CO has been shown at concentrations above 10% carboxyhaemoglobin. This study was undertaken to elucidate the possible anti-inflammatory effects of CO at lower CO concentrations. Effects of CO administration on cytokine (TNF-alpha, IL-6, IL-1beta and IL-10) release were investigated in a porcine model in which a systemic inflammatory response syndrome was induced by endotoxin infusion. Endotoxin was infused in 20 anaesthetized and normoventilated pigs. Ten animals were targeted with inhaled CO to maintain 5% COHb, and 10 animals were controls. In the control group, mean pulmonary artery pressure increased from a baseline value of 17 mmHg (mean, n = 10) to 42 mmHg (mean, n = 10) following 1 hour of endotoxin infusion. Similar mean pulmonary artery pressure values were found in animals exposed to carbon monoxide. Plasma levels of all of the measured cytokines increased in response to the endotoxin infusion. The largest increase was observed in TNF-alpha, which peaked after 1.5 hours at 9398 pg/ml in the control group and at 13395 pg/ml in the carbon monoxide-exposed group. A similar peak was found for IL-10 while the IL-6 concentration was maximal after 2.5 hours. IL-1beta concentrations increased continuously during the experiment. There were no significant differences between carbon monoxide-exposed animals and controls in any of the measured cytokines. Our conclusion is that 5% COHb does not modify the cytokine response following endotoxin infusion in pigs.


Histocompatibility and other risk factors for histological rejection of human cardiac allografts during the first three months following transplantation.
A histological analysis of 2564 endomyocardial biopsies was conducted in 349 cardiac transplant patients to determine potential risk factors for acute cellular rejection during the first three months following transplantation. This analysis dealt with the frequency, time of onset, and duration of cellular rejection. Patients on perioperative RATG experienced significantly less rejection than patients on OKT3 or without antilymphocyte antibody immunoprophylaxis. A trend was noted toward increased rejection in recipients diagnosed originally with chronic myocarditis compared with patients in other disease categories including ischemic heart disease and dilated cardiomyopathy. No significant differences were seen in histological rejection between male and female recipients. On the other hand, patients over 55 years of age were found at lower risk of histological rejection. The results of this analysis have demonstrated quite clearly, but not unexpectedly, that a greater degree of HLA mismatching correlates with increased cellular rejection. This effect was noted not only for the HLA-A,B and DR antigens, but also HLA-DQ and HLA-DRw52/53 antigens. In multivariate analysis, the highest level of statistical significance was obtained for the combined HLA-A,B,DR and DQ group. Sensitized patients with panel-reactive lymphocytotoxic antibodies of greater than 10% experienced more histological rejection than nonsensitized patients. On the other hand, a positive lymphocytotoxic crossmatch did not appear to influence cellular rejection of cardiac allografts. Also, no differences were seen in histological rejection between ABO-identical and compatible heart transplants. These findings further support the concept that donor HLA compatibility and pretransplant sensitization represent significant risk factors for cellular rejection in cardiac transplantation.


Determination of disulfide array and subunit structure of taste-modifying protein, miraculin.
The taste-modifying protein, miraculin (Theerasilp, S. et al. (1989) J. Biol. Chem. 264, 6655-6659) has seven cysteine residues in a molecule composed of 191 amino acid residues. The formation of three intrachain disulfide bridges at Cys-47-Cys-92, Cys-148-Cys-159 and Cys-152-Cys-155 and one interchain disulfide bridge at Cys-138 was determined by amino acid sequencing and composition analysis of cystine-containing peptides isolated by HPLC. The presence of an interchain disulfide bridge was also supported by the fact that the cystine peptide containing Cys-138 showed a negative color test for the free sulfhydryl group and a positive test after reduction with dithiothreitol. The molecular mass of non-reduced miraculin (43 kDa) in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was nearly twice the calculated molecular mass based on the amino acid sequence and the carbohydrate content of reduced miraculin (25 kDa). The molecular mass of native miraculin determined by low-angle laser light scattering was 90 kDa. Application of a crude extract of miraculin to a Sephadex G-75 column indicated that the taste-modifying activity appears at 52 kDa. It was concluded that native miraculin in pure form is a tetramer of the 25 kDa-peptide and native miraculin in crude state or denatured, non-reduced miraculin in pure form is a dimer of the peptide. Both tetramer miraculin and native dimer miraculin in crude state had the taste-modifying activity.


You are who your friends are-nuclear pore proteins as components of chromatin-binding complexes.
Nuclear pore complexes are large multicomponent protein complexes that are embedded in the nuclear envelope, where they mediate nucleocytoplasmic transport. In addition to supporting transport, nuclear pore components, termed nucleoporins (Nups), can interact with chromatin and influence genome function. A subset of Nups can also localize to the nuclear interior and bind chromatin intranuclearly, providing an opportunity to investigate chromatin-associated functions of Nups outside of the transport context. This review focuses on the gene regulatory functions of such intranuclear Nups, with a particular emphasis on their identity as components of several chromatin regulatory complexes. Recent proteomic screens have identified Nups as interacting partners of active and repressive epigenetic machinery, architectural proteins, and DNA replication complexes, providing insight into molecular mechanisms via which Nups regulate gene expression programs. This review summarizes these interactions and discusses their potential functions in the broader framework of nuclear genome organization.


P(700) Chlorophyll a-Protein : Purification, Characterization, and Antibody Preparation.
The P(700) chlorophyll alpha-protein was purified by preparative sodium dodecyl sulfate (SDS) gel electrophoresis from SDS-solubilized barley (Hordeum vulgare L., cv Himalaya) chloroplast membranes. After elution from the gel in the presence of 0.05 to 0.1% Triton X-100, the recovered protein had a chlorophyll/P(700) ratio of 50 to 60/1 and contained no chlorophyll b or cytochromes. Analysis of the polypeptide composition of the chlorophyll-protein revealed a 58 to 62 kilodalton (kD) polypeptide component but no lower molecular weight polypeptides. The 58 to 62 kD component was further resolved into two distinct polypeptide bands which were subsequently mapped by partial cyanogen bromide digestion and Staphylococcus aureus proteolysis. Based on results from the mapping experiments and other data, we suggest that the two components are conformational variants of a single polypeptide. Measurement of the chlorophyll to protein ratio by quantitative amino acid analysis and consideration of the yield of P(700) in the protein isolate suggest that, contrary to previous models (Bengis and Nelson, 1975, 1977), P(700)in vivo is associated with a minimum of four subunits of approximately 60 kD.Antibodies raised against the photochemically active chlorophyll-protein complex from barley reacted specifically with the 58 to 62 kD apoprotein. The same preparative electrophoresis procedure was used to isolate photochemically active P(700) chlorophyll a-protein from soybean (Glycine max L.), tobacco (Nicotiana tobacum L.), petunia (Petunia x hybrida), tomato (Lycopersicum esculentum), and Chlamydomonas reinhardti. The isolated complex from all species exhibited identical polypeptide compositions and chlorophyll/P(700) ratios. Antibodies to the barley protein cross reacted with all species tested demonstrating the highly conserved structure of the apoprotein.


Expression of MUC1 on corneal endothelium of human.
To determine the expression of MUC1 in the human corneal endothelium. Reverse transcription-polymerase chain reaction (RT-PCR) for MUC1 was performed with total RNA from endothelial cells extracted from the human cornea. In situ hybridization with sense and antisense probes of human MUC1 was performed on the human corneal endothelium, immunoblot analysis using monoclonal antibody specific for human MUC1 (HMFG-1, or VU4H5) was performed on collected human corneal endothelial cells, and immunohistochemistry on the human cornea, using the same antibodies. MUC1 mRNA expression was observed by RT-PCR in the human corneal endothelium, and the nucleotide sequence from the amplified band was matched with known human MUC1. In situ hybridization studies showed the localization of MUC1 mRNA in the human corneal endothelium, and immunoblot assay demonstrated the presence of MUC1 protein (MW > 200 kd). In addition, MUC1 protein was observed on the apical surface of cells and at the superficial layer of the cytoplasm in immunohistochemical studies. Human corneal endothelial cells produce MUC1, which is known to have protective and lubricative roles.


Characterization of Candida parapsilosis infection of an in vitro reconstituted human oral epithelium.
Oral candidosis is a common problem in immunocompromised patients, and whilst Candida albicans is regarded as the principal cause of infection, other non-Candida albicans Candida (NCAC) species are increasingly being recognized as human pathogens. Relatively little is known about the virulence factors associated with NCAC species, and the aim of this study was to use a reconstituted human oral epithelium (RHOE) to examine epithelial infection withCandida parapsilosis. Strains originating from the oral and vaginal mucosa and from the urinary tract were all shown to colonize RHOE in a strain-dependent manner. Strain differences were found in the colonizing morphology and in the extent of invasion of the RHOE. Low invasion of RHOE was detected for strains after 12 h, whereas extensive tissue damage was evident after 24 h when assessed using histological examination and lactate dehydrogenase activity determination. Tissue damage was reduced in the presence of pepstatin A, although C. parapsilosis invasion of the tissue was not inhibited. Real-time polymerase chain reaction of secreted aspartyl proteinase (SAP) genes (SAPP1-3) showed that expression was strain dependent, with an increased expression generally occurring for Candida infecting RHOE compared with planktonic equivalents. In summary, C. parapsilosis was not highly invasive of RHOE but did induce significant tissue damage, which could relate to specific SAPgene expression.


Induction of enzymes involved in DNA repair and mutagenesis.
Three phenomena examined here have been claimed to reflect the operation of inducible repair systems. It has been postulated that 'SOS repair' involves the induction by DNA-damaging agents of an error-prone repair system that is capable of affecting the replication of damaged DNA. This system works with bacteriophages and animal viruses, in which it is possible to separate the effects of DNA damage on the viral DNA from that on the host cell. Whether this system also operates in repair of the cell's own DNA is, however, controversial. The system appears to have little effect on survival of bacterial cells and its operation in cellular mutagenesis is still not proven, at least in bacteria with otherwise normal repair capacity. 'Adaptation' is the response of bacteria to low doses of methylating agents. Adapted bacteria are more resistant to the lethal and mutagenic action of alkytlating agents. The process includes the induction of an enzyme that 'removes' O6-alkylated bases from DNA and which, unusually, is consumed during the course of the reaction. The reaction itself is also unknown; it does not depend on a nuclease, glycosylase or demethylase, but could use a transmethylase. There is some evidence that an analogous process occurs in animals. The third process affects the synthesis of high molecular weight DNA in cultured mammalian cells that have been exposed to split doses of DNA-damaging agents. It has been postulated that this system is inducible and error-free, but detailed analysis suggests that the observed effect is an artifact arising from an abnormal distribution of sizes of nascent DNA after the second dose, and as a result of exposure to the first dose. Inducible DNA repair systems may be expected to influence kinetics of the dose--response relationships obtained after exposure of cells to mutagens and carcinogens. The interactions between these effects and those produced by inducible pathways of metabolic activation and detoxification are discussed.


[The evaluation of cerebral function by diffusion weighted imaging after norepinephrine-induced hypertensive perfusion therapy in pig model of cardiac arrest].
To evaluate the changes in cerebral function of pigs with cardiac arrest (CA) after recovery of spontaneous circulation (ROSC) after hypertension perfusion therapy induced by norepinephrine (NE). Ventricular fibrillation (VF) was induced by electrical stimulation, and standard cardiopulmonary resuscitation (CPR) was performed after VF for 4 minutes. The pigs with successful ROSC were randomly divided into two group, each n=5. The pigs in the hypertensive reperfusion group were given with NE immediately to maintain the mean arterial pressure (MAP) at 130% before VF for 4 hours; MAP of the pigs in normal reperfusion group was maintained for 4 hours as baseline. The changes in hemodynamics were observed for 4 hours in both groups. Cerebral cortex was scanned with diffusion weighted imaging (DWI) before VF and 1 hour and 3 hours after ROSC, and the dynamic changes in brain functional imaging were observed. Twenty-four hours after ROSC, brain biopsy were collected and examined after hematoxylin and eosin staining (HE). Compared with the normal reperfusion group, heart rate (HR), MAP, cardiac output (CO) and coronary perfusion pressure (CPP) in the hypertensive reperfusion group showed a tendency to increase (ROSC 30 minutes HR: 167±8 bpm vs. 140±15 bpm, ROSC 1 hour MAP: 131±9 mm Hg vs. 108±10 mm Hg, ROSC 1 hour CO: 4.9±0.1 L/min vs. 3.4±0.5 L/min, ROSC 2 hours CPP: 118±12 mm Hg vs. 88±1 mm Hg, P<0.05 or P<0.01). There was no obvious abnormality as shown by DWI before and after resuscitation, and the apparent diffusion coefficient (ADC) showed a tendency to decrease after resuscitation in both groups. The ADC in the normal reperfusion group was decreased more than that in the hypertensive reperfusion. Pathological study showed that the protective effect of the hypertensive reperfusion on brain tissue was better than that of the normal reperfusion group. Hypertensive reperfusion can produce hemodynamic changes, and an increase in cerebral blood flow, thus it produces a protective effect on brain to promote the recovery of neurological function in pigs with CA after resuscitation.


Unilateral Nephrectomy Stimulates ERK and Is Associated With Enhanced Na Transport.
Nephron loss initiates compensatory hemodynamic and cellular effects on the remaining nephrons. Increases in single nephron glomerular filtration rate and tubular flow rate exert higher fluid shear stress (FSS) on tubules. In principal cell (PC) culture models FSS induces ERK, and ERK is implicated in the regulation of transepithelial sodium (Na) transport, as well as, proliferation. Thus, we hypothesize that high tubular flow and FSS mediate ERK activation in the cortical collecting duct (CCD) of solitary kidney which regulates amiloride sensitive Na transport and affects CCD cell number. Immunoblotting of whole kidney protein lysate was performed to determine phospho-ERK (pERK) expression. Next, sham and unilateral nephrectomized mice were stained with anti-pERK antibodies, and dolichos biflorus agglutinin (DBA) to identify PCs with pERK. Murine PCs (mpkCCD) were grown on semi-permeable supports under static, FSS, and FSS with U0126 (a MEK1/2 inhibitor) conditions to measure the effects of FSS and ERK inhibition on amiloride sensitive Na short circuit current (Isc). pERK abundance was greater in kidney lysate of unilateral vs. sham nephrectomies. The total number of cells in CCD and pERK positive PCs increased in nephrectomized mice (9.3 ± 0.4 vs. 6.1 ± 0.2 and 5.1 ± 0.5 vs. 3.6 ± 0.3 cell per CCD nephrectomy vs. sham, respectively, n > 6 per group, p < 0.05). However, Ki67, a marker of proliferation, did not differ by immunoblot or immunohistochemistry in nephrectomy samples at 1 month compared to sham. Next, amiloride sensitive Isc in static mpkCCD cells was 25.3 ± 1.7 μA/cm2 (n = 21), but after exposure to 24 h of FSS the Isc increased to 41.4 ± 2.8 μA/cm2 (n = 22; p < 0.01) and returned to 19.1 ± 2.1 μA/cm2 (n = 18, p < 0.01) upon treatment with U0126. Though FSS did not alter α- or γ-ENaC expression in mpkCCD cells, γ-ENaC was reduced in U0126 treated cells. In conclusion, pERK increases in whole kidney and, specifically, CCD cells after nephrectomy, but pERK was not associated with active proliferation at 1-month post-nephrectomy. In vitro studies suggest high tubular flow induces ERK dependent ENaC Na absorption and may play a critical role in Na balance post-nephrectomy.


[Tissue-specific regulation of rat liver mitochondrial oxidative phosphorylation by a soluble phase of cells from that organ].
Tissue specificity of mitochondrial respiration stimulation under the effect of a soluble phase of liver cells (SPC) is preserved by addition to dinitrophenol but is reserved in the presence of oligomycin. Addition of rotenon in the presence of SPC entails a tissue-specific increase in respiration that is proportional to the respective increase in respiration of intact mitochondria in the presence of the inhibitor mentioned. SPC tissue-specifically inhibits ATPase activity of liver mitochondria. This fraction of SPC is capable of recovering the coupling of oxidative phosphorylation of mitochondria whose respiration is inhibited by adding ADP. A conclusion is made that SPC is capable not only to decrease tissue-specifically the coupling of intact mitochondria but also to raise it in mitochondria with deranged oxidative phosphorylation. This assures intratissue organization of liver metabolism by means of tissue-specific stabilization of liver cell energy metabolism.


Multimodal strategies as prerequisite for organ-salvaging tumor surgery.
Curative therapy for solid tumors often requires combined-modality treatment. In the following paper the importance of different strategies as prerequisite for organ salvaging tumor surgery and for successful treatment will be shown. Unfavorable surgical conditions (e.g. site, extent of the tumor) often result in nonresectability or mutilating surgery. Preoperative therapies must try to achieve a maximum tumor shrinkage aiming at attaining resectability and preservation of organ function and life. The exact evaluation of the response to preoperative therapy is important since the degree of the response correlates directly with outcome. Unfavorable response to preoperative therapy means unfavorable prognosis and requires further, more aggressive therapy. A very important parameter in this evaluation is the "histologic response", measured by the degree of tumor devitalization at the time of definitive surgery. However, even in cases with complete clinical and histological remission, complete resection of the tumor after preoperative treatment is essential. Despite good response to preoperative treatment followed by complete resection, therapy must be sufficiently continued after surgery. Preoperative therapy should achieve maximum tumor control, that means total necrosis of all tumor cells to make a complete resection of the devitalized tumor easier and to get the possibility to preserve organ function.


High-yielding enzymatic alpha-glucosylation of pyridoxine by marine alpha-glucosidase from Aplysia fasciata.
We recently succeeded in the identification and purification of an interesting marine exo-alpha-glucosidase (EC 126.96.36.199) from the anaspidean mollusc Aplysia fasciata. The enzyme was characterized by good transglycosylation activity toward different acceptors using maltose as donor. High-yielding enzymatic alpha-glycosylation of pyridoxine using this marine enzyme is reported here; the reaction has been optimized, reaching 80% molar yield of products (pyridoxine monoglucosides 24 g/l; pyridoxine isomaltoside 35 g/l). High selectivity toward the 5' position is observed for both monoglucoside and disaccharide formation. This is the first report describing the enzymatic production of pyridoxine isomaltoside.


[Two Cases in Which Eribulin Mesylate Was Effective for Taxane-Resistant Advanced Breast Cancer].
We experienced 2 cases in which eribulin mesylate was effective for taxane-resistant advanced breast cancer. Case 1: A 65- year-old woman was diagnosed with advanced breast cancer(T4cN2aM0, stage III B)and treated with chemotherapy(nabpaclitaxel). Bone metastasis was observed; then, she sequentially received epirubicin and cyclophosphamide, nab-paclitaxel, and bevacizumab and paclitaxel. However, lung metastases appeared, and we changed the regimen to eribulin mesylate. We administered 11 courses of eribulin mesylate before bone marrow metastasis appeared. Eribulin mesylate was effective for more than 1 year. Case 2: A 77-year-old woman was diagnosed with advanced breast cancer(T4bN3cM1, stage IV), and liver and pleural metastasis were observed during an examination at the first visit. Four courses of nab-paclitaxel were administered, but because of the increase in pleural effusion, we changed the regimen to eribulin mesylate. Thirteen courses of eribulin mesylate were administered before the disease progressed. The progression-free survival was 9.53 months. Through the 2 cases in which eribulin mesylate was effective for taxane-resistant advanced breast cancer, the effect of eribulin mesylate compared to taxane was clinically inferred.


Phlogistic properties of peptidoglycan-polysaccharide polymers from cell walls of pathogenic and normal-flora bacteria which colonize humans.
PG-PS polymers which can induce experimental chronic inflammation in joints and other tissues can be isolated from the cell walls of human pathogens, such as group A streptococci, as well as from certain indigenous bacterial species which colonize the human intestinal tract. The structural and biological properties that are required for cell wall fragments to express this remarkable activity are still not well defined, but polymer size, resistance to tissue enzymes, and capacity to sustain activation of complement, macrophages, neutrophils, and T cells are properties associated with the most active preparations. There is increasing evidence that PG-PS structures with arthropathogenic activity occur in the human intestinal lumen and that these polymers can be translocated systemically. These observations support the concept that PG-PS, derived from a variety of bacterial species, can be part of the etiology of rheumatoid arthritis and other chronic inflammatory diseases. Since the PG component provides a common element to which all individuals are exposed, it follows that susceptibility is related to efficiency of disposal of bacterial cell wall debris, as well as to cytokine networks and immune cell function (51).


Genistein anticancer efficacy during induced oral squamous cell carcinoma: an experimental study.
About 7 million people die from various types of cancer every year representing nearly 12.5% of deaths worldwide. This fact raises the demand to develop new, effective anticancer, onco-suppressive, and chemoprotective agents for the future fighting of cancers. Genistein exhibits pleiotropic functions in cancer, metabolism, and inflammation. It functions as an antineoplastic agent through its effect on the cell cycle, apoptotic processes, angiogenesis, invasion, and metastasis. The current study aimed to study the genistein onco-suppressive effects during 7,12-dimethylbenz[a]anthracene (DMBA)-induced oral carcinogenesis in hamsters' buccal pouch utilizing flow cytometry analysis (FMA), as a fast-diagnosing tool, in addition to the histopathology. The buccal mucosa of adult male Syrian hamsters was painted with paraffin oil only (group 1), DMBA mixed in mineral oil (group 2), or orally administrated genistein along with painting DMBA (group 2B). The buccal mucosa was utilized for flow cytometric analysis and histopathological examination. Grossly, DMBA-induced carcinogenesis started at the 9th week. Progressive signs appeared in the following weeks reaching to large ulcerative oral masses and exophytic nodules at the 21st week. Histologically, invasive well-differentiated oral squamous cell carcinoma (OSCC) appeared in the underlying tissues from the 12th week, showing malignant criteria. Genistein had delayed clinicopathological change, which started 6 weeks later, than the DMBA-painted hamsters, as mild epithelial dysplastic changes. This became moderate during the last 6 weeks, without dysplastic changes. Flow cytometry revealed that DMBA led to considerable variation in DNA proliferation activity, aneuploid DNA pattern, in 47.22% of hamsters and significantly raised the S-phase fragment (SPF) values, which drastically reduced after genistein treatment. Taken together, genistein could be employed as an onco-suppressive agent for carcinogenesis. Moreover, FMA could be used as an aiding fast tool for diagnosis of cancer.


Clinical significance of age-related changes of the palpebral fissures between age 2 and 18 years in healthy Caucasians.
The development of surface measurements of the palpebral fissures and age-related changes in the quality of the relationships between the individual measurements were followed in 1552 healthy Caucasians between ages 2 and 18. At age 2, the height of the palpebral fissure and the biocular width (ex-ex) were the most developed features (93.3% and 86%) and the least developed was the intercanthal width (77.6% to 82.9%). The measurements reached adult size between ages 8 (intercanthal width in girls) and 16 (palpebral fissure inclination in boys). The rate of growth in the orbital measurements was usually moderate, seldom above-average and fast only in intercanthal width between ages 3 and 4. The study determined the periods with minimal growth (approximately ages 5 to 7 and 9 to 10) for each of the measurements. After maturation, the changes in measurements were minimal. A knowledge of the developmental levels of the measurements at an early age, their changes with age and their maturation times are of great importance in timing early or final corrective surgical procedures.


Detection of a type IX collagen-related mRNA in an invertebrate, the marine annelid Nereis virens.
Fibrous and non-fibrous collagens have been described in both vertebrate and invertebrate animals. However, there has been limited characterization of non-fibrous collagens and their corresponding genes in invertebrate animals. In the present study we have used as a probe an avian cDNA clone which encompasses the COL3, NC3 and part of the COL2 domain of the collagen alpha 3(IX) subunit. This probe hybridized to mRNA obtained from the cuticle and body of the marine annelid, Nereis virens. Northern blot hybridization exhibited an mRNA of ca. 7.5-8 kilobases which in situ hybridization shows to be most abundant over cuticle-associated cells. Dot-blot hybridization, comparing cuticle mRNA and body mRNA, indicates that this collagen mRNA is five times more abundant in the cuticle. The composite data suggest evolutionary conservation, in both vertebrate and invertebrate animals, of a non-fibrillar collagen.


Induction of cellular immunity by immunization with novel hybrid peptides complexed to heat shock protein 70.
Heat shock proteins 70 (hsp70) derived from tissues and cells can elicit cytotoxic T lymphocyte (CTL) responses against peptides bound to hsp70. However, peptides can markedly differ in their affinity for hsp, and this potentially limits the repertoire of peptides available to induce CTL by the hsp immunization. Hybrid peptides consisting of a high-affinity ligand for the peptide-binding site of hsp70 joined to T cell epitopes by a glycine-serine-glycine linker were constructed. Immunization with hybrid peptides complexed to mouse hsp70 effectively primed specific CTL responses in mice and were more potent than T cell peptide epitopes alone with hsp70. In vivo immunization with hsp70 and hybrid peptides led to rejection of tumors expressing antigen with greater efficacy than immunization with peptide epitope plus hsp70. Induction of CTL responses occurred independently of CD4(+) T cells, suggesting that immunization directly primed antigen-presenting cells to elicit CD8(+) cytotoxic T cell responses without T cell help. Both peptide/hsp70 complexes and mouse hsp70 alone were able to induce cultures of mouse bone marrow-derived dendritic cells (DC) to release cytokines, including DC from endotoxin-resistant C57BL/10Sc mice. Thus, hsp70/hybrid peptide complexes can activate DC for cytokine release, providing a potential adjuvant effect that could bypass T cell help.


Applications of green fluorescent protein in plants.
Green fluorescent protein (GFP) is increasingly being used in plant biology from the cellular level to whole plant level. At the cellular level, GFP is being used as an in vivo reporter to assess frequency of transient and stable transformation. GFP has also proven to be an invaluable tool in monitoring trafficking and subcellular localization of protein. At the organ level and up, many exciting applications are rapidly emerging. The development of brighter GFP mutants with more robust folding properties has enabled better macroscopic visualization of GFP in whole leaves and plants. One interesting example has been the use of GFP to monitor virus movement in and among whole plants. GFP is also emerging as a powerful tool to monitor transgene movement and transgenic plants in the field. In a proof-of-concept study, tobacco was transformed with a modified version of the GFP gene controlled by a constitutive (35S) promoter. GFP expression in progeny plants ranged from 0% to 0.5%, and approximately 0.1% GFP was the minimal amount needed for unambiguous macroscopic detection. GFP is the first truly in vivo reporter system useful in whole plants, and we project its usefulness will increase even further as better forms of GFP genes become available.


Cycling efficiency in trained male and female competitive cyclists.
The aim of this study was to examine differences in cycling efficiency between competitive male and female cyclists. Thirteen trained male (mean ± SD: 34 ± 8 yr, 74.1 ± 6.0 kg, Maximum Aerobic Power (MAP) 414 ± 40 W, VO2max 61.3 ± 5.4 ml·kg(-1)·min(-1)) and 13 trained female (34 ± 9 yr, 60.1 ± 5.2 kg, MAP 293 ± 22 W, VO2max 48.9 ± 6.1 ml·kg(-1)·min(-1)) competitive cyclists completed a cycling test to ascertain their gross efficiency (GE). Leg and lean leg volume of all cyclists was also measured. Calculated GE was significantly higher in female cyclists at 150W (22.5 ± 2.1 vs 19.9 ± 1. 8%; p < 0.01) and 180W (22.3 ± 1.8 vs 20.4 ± 1.5%; p = 0.01). Cadence was not significantly different between the groups (88 ± 6 vs 91 ± 5 rev·min(-1)). Lean leg volume was significantly lower for female cyclists (4.04 ± 0.5 vs 5.51 ± 0.8 dm(3); p < 0.01) and was inversely related to GE in both groups at 150 and 180W (r = -0.59 and -0.58; p < 0.05). Lean leg volume was shown to account for the differences in GE between the males and females. During an "unloaded "pedalling condition, male cyclists had a significantly higher O2 cost than female cyclists (1.0 ± 0.1 vs 0.7 ± 0.1 L·min(-1); p < 0.01), indicative of a greater non-propulsive cost of cycling. These results suggest that differences in efficiency between trained male and female cyclists can be partly accounted for by sex-specific variation in lean leg volume. Key pointsDifferences in GE exist between male and female cyclists.Males have a higher oxygen cost of "unloaded "cycling, as predicted by the intercept of the O2 cost-power output relationshipThis suggests that in addition to work rate, leg volume/mass may be an important determinant of observed differences in oxygen cost and therefore GE, between male and female competitive cyclists.


[qSOFA score for prediction of outcome in surgical patients in intensive care units. Post hoc analysis of the Russian multi-center trial Rises].
To define the informative value of qSOFA score in the prediction of outcomes in surgical patients admitted to the Intensive Care Units. Post hoc analysis of multicenter prospective observational study RISES. The following patient information was analyzed: gender, age, diagnosis, presence of infection, SIRS criteria, qSOFA and SOFA scores, outcomes. The study included data of 331 patients with surgical diseases. Infection was not observed in 174 (52.6%) cases, 157 (47.4%) patients had infection. In the group of patients without infection, area under ROC-curve for SIRS criteria was 0.519 (95% CI 0.429-0.610) and similar to that qSOFA (p=0.535). Area under ROC-curve for SOFA scale was 0.619 (95% CI 0.511-0.726) and did not significantly differ from this value for QSOFA (p=0.241). In the group of surgical patients with infection, area under ROC-curve for SIRS was 0.490 (95% CI 0.419-0.561), that was significantly lower than area under ROC-curve for qSOFA (p=0.016). Area under ROC-curve for SOFA scale was 0.803 (95% CI 0.681-0.924), that significantly exceeded area under ROC-curve for qSOFA (p=0.017). qSOFA scale is important in surgical patients with infection admitted to ICUs. Increased qSOFA score is associated with augmentation of mortality rate. qSOFA scale significantly exceeds the SIRS criteria, but is inferior to the SOFA in the prognosis of mortality in these patients.


Cell fate-specific regulation of EGF receptor trafficking during Caenorhabditis elegans vulval development.
By controlling the subcellular localization of growth factor receptors, cells can modulate the activity of intracellular signal transduction pathways. During Caenorhabditis elegans vulval development, a ternary complex consisting of the LIN-7, LIN-2 and LIN-10 PDZ domain proteins localizes the epidermal growth factor receptor (EGFR) to the basolateral compartment of the vulval precursor cells (VPCs) to allow efficient receptor activation by the inductive EGF signal from the anchor cell. We have identified EGFR substrate protein-8 (EPS-8) as a novel component of the EGFR localization complex that links receptor trafficking to cell fate specification. EPS-8 expression is upregulated in the primary VPCs, where it creates a positive feedback loop in the EGFR/RAS/MAPK pathway. The membrane-associated guanylate kinase LIN-2 recruits EPS-8 into the receptor localization complex to retain the EGFR on the basolateral plasma membrane, and thus allow maximal receptor activation in the primary cell lineage. Low levels of EPS-8 in the neighboring secondary VPCs result in the rapid degradation of the EGFR, allowing these cells to adopt the secondary cell fate. Extracellular signals thus regulate EGFR trafficking in a cell type-specific manner to control pattern formation during organogenesis.


Does vowel inventory density affect vowel-to-vowel coarticulation?
This study tests the output constraints hypothesis that languages with a crowded phonemic vowel space would allow less vowel-to-vowel coarticulation than languages with a sparser vowel space to avoid perceptual confusion. Mandarin has fewer vowel phonemes than Cantonese, but their allophonic vowel spaces are similarly crowded. The hypothesis predicts that Mandarin would allow more coarticulation than Cantonese. Eight native speakers of Cantonese and of Beijing Mandarin were recorded saying the target sequences /pV1 1pV2pV3/ (V = /i a u/) in carrier phrases. F1 and F2 frequencies were measured at vowel edge and midpoint, and were normalized for analyses. The results show that Cantonese and Mandarin do not differ in degree of vowel-to-vowel coarticulation in either F1 or F2. In addition, unstressed vowels exhibit more coarticulation than stressed vowels. Carryover coarticulation exceeds anticipatory coarticulation in both F1 and F2. Unstressed vowels in the carryover position are the most susceptible to coarticulation. The results show that vowel inventory does not predict vowel-to-vowel coarticulation. Fundamental assumptions of the output constraints hypothesis are evaluated to explain its failure in predicting language-specific patterns of vowel-to-vowel coarticulation. The importance of syntagmatic relationships in coarticulation is also discussed.


Comparison of two mammalian surveys made with camera traps in southeastern Brazil, focusing the abundance of wild mammals and domestic dogs.
Sampling allows assessing the impact of human activities on mammal communities. It is also possible to assess the accuracy of different sampling methods, especially when the sampling effort is similar. The present study aimed at comparing two mammalian surveys carried out over a three-year interval, in terms of sampling effort, capture success, abundance of domestic dogs, impact of human activities, and relative biomass using camera traps, in the Serra do Japi Biological Reserve and surroundings, located in Jundiaí, state of São Paulo, southeastern Brazil. The total richness recorded was 13 species, one domestic and 12 wild mammals. Sampling effort in both surveys was similar, but capture success and number of captures differed. The abundance of wild mammals and dogs did also differ between surveys. There was a highly significant correlation between abundance of wild mammals and capture effort for the survey performed in 2006/2007, but not for the survey performed in 2009/2010. The difference between samples may be related to human disturbance, since the number of domestic mammals photographed was higher in the second survey, three years after the first survey. Despite being a reserve, the area is still under pressure from urbanization, biological invasion, environmental degradation, and hunting, which may reduce the abundance of wild mammals.


Making the Most of Midlines: A Retrospective Review of Outcomes.
Decision-making for vascular access device selection is becoming increasingly complex as new technologies come to market and efforts to reduce central line-associated bloodstream infections increase. This retrospective review of 165 midline catheter outcomes was undertaken after a cluster of unexpected failures occurred in a large academic medical center in the southeastern United States. Mean dwell time for midline catheters was 8.5 days; 62.8% lasted to therapy completion, and complications occurred in 15.8%. A quality improvement initiative including implementation of a blood return algorithm and standard education for unit staff reduced infiltration and thrombosis complications.


Mollicutes-wall-less bacteria with internal cytoskeletons.
The structure and motility of the Mollicutes (Spiroplasma, Mycoplasma, and Acholeplasma) are briefly reviewed. The data are presented from the perspective of prokaryotic and eukaryotic motors, cytoskeletons, and cell motility. The Mollicutes are eubacteria derived from Clostridia by regressive evolution and genome reduction to produce the smallest and simplest free-living and self-replicating cells. Structurally, the Mollicutes are characterized by a complete lack of a cell wall and the presence of an internal cytoskeleton. Spiroplasma, which are helical cells with a flat, ribbon-like cytoskeleton, are amenable to structural and geometrical analysis. Motility and shape changes can be explained and modeled by the cytoskeleton acting as a linear motor.


Portable fluorescence lifetime spectroscopy system for in-situ interrogation of biological tissues.
Fluorescence spectroscopy and lifetime techniques are potential methods for optical diagnosis and characterization of biological tissues with an in-situ, fast, and noninvasive interrogation. Several diseases may be diagnosed due to differences in the fluorescence spectra of targeted fluorophores, when, these spectra are similar, considering steady-state fluorescence, others may be detected by monitoring their fluorescence lifetime. Despite this complementarity, most of the current fluorescence lifetime systems are not robust and portable, and not being feasible for clinical applications. We describe the assembly of a fluorescence lifetime spectroscopy system in a suitcase, its characterization, and validation with clinical measurements of skin lesions. The assembled system is all encased and robust, maintaining its mechanical, electrical, and optical stability during transportation, and is feasible for clinical measurements. The instrument response function measured was about 300 ps, and the system is properly calibrated. At the clinical study, the system showed to be reliable, and the achieved spectroscopy results support its potential use as an auxiliary tool for skin diagnostics.


Calcium-dependent binding of annexin 12 to phospholipid bilayers: stoichiometry and implications.
Annexins (ANXs) are a superfamily of proteins whose functional hallmark is Ca2+-dependent binding to anionic phospholipids. Their core domains are usually composed of a 4-fold repeat of a conserved amino acid sequence, with each repeat containing a type II Ca2+ binding site that is generally thought to mediate Ca2+-dependent binding to the membrane. We now report that ANX12 binding to phospholipid vesicles is highly cooperative with respect to Ca2+ concentration (Hill constant approximately 7), thereby suggesting that more than the four well-characterized type II Ca2+ binding sites are involved in phospholipid binding. Two independent approaches, a novel 45Ca2+ copelleting assay and isothermal titration calorimetry, indicate a stoichiometry of approximately 12 mol of Ca2+/mol of ANX12 for binding to phospholipid vesicles. On the basis of the "low-affinity" Ca2+-binding sites in a number of ANX X-ray crystal structures, we propose a model for ANX12 bilayer binding that involves three types of Ca2+ sites in each of the four repeats. In this model, there is a complementarity between the spacing of the ANX12 Ca2+ binding sites and the spacing of the phospholipid headgroups in bilayers. We tested the implications of the model by manipulating the physical state of vesicles composed of phospholipids with saturated acyl chains with temperature and measuring its influence on ANX12 binding. ANX12 bound to vesicles in a Ca2+-dependent manner when the vesicles were in the liquid crystal phase but not when the phospholipid was in the gel phase. Furthermore, ANX12 bound initially to fluid bilayers remained bound when cooled to 4 degrees C, a temperature that should induce the gel phase transition. Overall, these studies suggest that ANX12 is well suited to being a Ca2+ sensor for rapid all-or-none intercellular membrane-related events.


Lesions of the adrenal cortex.
In surgical pathology practice adrenal cortical tumors are rare. However, in autopsy series adrenal cortical nodules are found frequently. These are now being identified more commonly in life when the abdomen is scanned for other disease. It is important to differentiate between benign and malignant lesions as adrenal cortical carcinoma is an aggressive tumor. Molecular genetic investigations are providing new information on both pathogenesis of adrenal tumors and basic adrenal development and physiology. To provide an overview of current knowledge on adrenal cortical development and structure that informs our understanding of genetic diseases of the adrenal cortex and adrenal cortical tumors. Literature review using PubMed via the Endnote bibliography tool. The understanding of basic developmental and physiologic processes permits a better understanding of diseases of the adrenal cortex. The information coming from investigation of the molecular pathology of adrenal cortical tumors is beginning to provide additional tests for the assessment of malignant potential in diagnosis but the mainstay remains traditional histologic analysis.


Feasibility and Safety of a Novel Leg Exercise Apparatus for Venous Thromboembolism Prophylaxis after Total Joint Arthroplasty of the Lower Extremities-A Pilot Study.
Venous thromboembolism (VTE) is a severe complication in orthopedic surgeries. Herein, we developed a novel leg exercise apparatus (LEX) to encourage postoperative limb movement in bedridden patients to prevent VTE. We aimed to evaluate its feasibility and safety in individuals at risk of VTE. Twenty patients (four men, 16 women) who underwent total joint arthroplasty in the lower extremity were enrolled in this prospective study. Exercise using the LEX was performed for 5 min at 30 cycles/min, four times/day during postoperative days 1-7. Clinical assessments included the evaluation of vital signs, venous ultrasonography, and blood tests within seven days postoperatively, and adverse events (pulmonary embolism and cerebral hemorrhage) were monitored. Overall, 16/20 (80%) patients completed the 7-day exercise regimen. There were no cases of severe adverse events, changes in vital signs, or lower-extremity deep vein thrombosis in patients who performed exercises with the LEX. Thus, the results of this pilot study show that this novel apparatus may be a safe and feasible tool for VTE prophylaxis after joint arthroplasty of the lower extremities.


Exclusion of the uniform tetraploid cells significantly improves specificity of the urine FISH assay.
The urine fluorescence in situ hybridization (FISH) assay (UroVysion™), with the current scoring criteria, has a higher sensitivity than routine cytopathology but a lower specificity. Among 215 urine FISH tests we performed, 45 had associated histopathology and clinical follow up. In this study, a cell with four signals for each probe was classified as a uniform tetraploid cell (UTC); a presumed reparative cell which is currently classified as an abnormal cell in the FDA approved assay. By using the existing criteria, the tests were scored as positive or negative before and after exclusion of the UTCs. Before the exclusion, 24 positive, 13 negative, seven false positive, and one false negative result were obtained with 96% sensitivity and 65% specificity. After the exclusion, the results changed to 22 positive, 19 negative, one false positive, and three false negatives resulting in a 88% sensitivity of 88% and a 95% specificity; a significant improvement in the specificity. We conclude that exclusion of the UTCs as abnormal cells would result in a more solid performance of the FISH assay.


Effects of syllable stress in adaptation to altered auditory feedback in vowels.
Unstressed syllables in English most commonly contain the vowel quality [ə] (schwa), which is cross-linguistically described as having a variable target. The present study examines whether speakers are sensitive to whether their auditory feedback matches their target when producing unstressed syllables. When speakers hear themselves producing formant-altered speech, they will change their motor plans so that their altered feedback is a better match to the target. If schwa has no target, then feedback mismatches in unstressed syllables may not drive a change in production. In this experiment, participants spoke disyllabic words with initial or final stress where the auditory feedback of F1 was raised (Experiment 1) or lowered (Experiment 2) by 100 mels. Both stressed and unstressed syllables showed adaptive changes in F1. In Experiment 1, initial-stress words showed larger adaptive decreases in F1 than final-stress words, but in Experiment 2, stressed syllables overall showed greater adaptive increases in F1 than unstressed syllables in all words, regardless of which syllable contained the primary stress. These results suggest that speakers are sensitive to feedback mismatches in both stressed and unstressed syllables, but that stress and metrical foot type may mediate the corrective response.


Ribozymes: applications to functional analysis and gene discovery.
Ribozymes are catalytic RNA molecules that cleave RNAs with high specificity. Since the discovery of these non-protein enzymes, the rapidly developing field of ribozymes has been of particular interest because of the potential utility of ribozymes as tools for reversed genetics. However, despite extensive efforts, the activity of ribozymes in vivo has not usually been high enough to achieve the desirable biological effects. Now, by the use of RNA polymerase III (pol III) promoters, the ribozyme activity in cells has been successfully improved by developing efficient transport systems for the transcripts to the cytoplasm. In addition, it is possible to cleave a specific target RNA in cells by using an allosterically controllable ribozyme or an RNA-protein hybrid ribozyme. These ribozymes are potentially applicable to molecular gene therapy and efficient gene discovery systems. Furthermore, the developed pol III expression system is applicable to the expression of small interfering RNAs (siRNAs). The advantage of such ribozymes over siRNAs is the high specificity of the ribozyme that would not cause interferon responses.


The role of endocannabinoids in the hypothalamic regulation of visceral function.
The hypothalamus plays an important role in the regulation of several visceral processes, including food intake, thermoregulation and control of anterior pituitary secretion. Endogenous cannabinoids and CB(1) cannabinoid receptors have been found in the hypothalamus. In the present review, we would like to clarify the role of the endocannabinoid system in the regulation of the above-mentioned visceral functions. There is historical support for the role of marihuana (i.e. exogenous cannabinoids) in the regulation of appetite. Endocannabinoids also stimulate food intake. Furthermore, the specific CB(1) receptor antagonist SR141716 reduces food intake. Leptin treatment decreases endocannabinoid levels in normal rats and ob/ob mice. These findings provide evidence for the role of the hypothalamic endocannabinoid system in food intake and appetite regulation. Cannabinoids can change body temperature in a dose-dependent manner. High doses cause hypothermia while low doses cause hyperthermia. Cannabinoid administration decreases heat production. It seems that the effects of can- nabinoids on thermoregulation is exerted by altering some neurochemical mediator effects at both the presynaptic and postsynaptic level.THC and endocannabinoids have mainly inhibitory effects on the regulation of reproduction. Administration of anandamide (AEA) decreases serum luteinizing hormone (LH) and prolactin (PRL) levels. AEA causes a prolongation of pregnancy in rats and temporarily inhibits the postnatal development of the hypothalamo-pituitary axis in offspring. The action of AEA on the reproductory parameters occurs at both the hypothalamic and pituitary level. CB(1) receptors have also been found in the anterior pituitary. Further, LH levels in CB(1) receptor-inactivated mice were decreased compared with wild-type mice. Taken together, all these observations suggest that the endocannabinoid system is playing an important part in the regulation of the mentioned visceral functions and it provides the bases for further applications of cannabinoid receptor agonists and/or antagonists in visceral diseases regulated by the hypothalamus.