Current and future treatment of amyloid neuropathies.
Amyloid neuropathies of acquired or genetic origin are disabling and life-threatening, until recently there were few treatment options available. Poor prognosis is related to progressive neuropathy and associated, although often underdiagnosed, cardiac involvement in specific transthyretin (TTR) gene mutations. Recent progress has modified prognosis and management of amyloid neuropathies. In TTR-familial amyloidosis with polyneuropathy, major changes have occurred over the last 30 years: better knowledge concerning genetics, phenotypes and epidemiology, and the advent of possible treatments. Liver transplantation, first performed in 1990, stopped disease progression, thus doubling survival in early onset V30M patients. More recently tetramer stabilizers (Tafamidis and Diflunisal) showed a significant reduction of progression of neuropathic scores; Tafamidis is now recommended in Stage I patients. Two multicentric clinical trials are now ongoing to evaluate TTR gene silencing by antisense Oligonucleotides (ASO) or siRNA. In the near future we should have new therapeutical options for patients with amyloid neuropathy.


"Click chemistry" in tailor-made polymethacrylates bearing reactive furfuryl functionality: a new class of self-healing polymeric material.
This investigation reports the effective use of the Diels-Alder (DA) reaction, a "click reaction" in the preparation of thermally amendable and self-healing polymeric materials having reactive furfuryl functionality. In this case, the DA and retro-DA (rDA) reactions were carried out between the tailor-made homo- and copolymer of furfuryl methacrylate prepared by atom-transfer radical polymerization and a bismaleimide (BM). The kinetic studies of DA and rDA reactions were carried out using Fourier transform infrared spectroscopy. The DA polymers were insoluble in toluene at room temperature. When the DA polymers were heated at 100 degrees C in toluene, it was soluble. This is because of the cleavage between furfuryl functionality and BM. The chemical cross-link density was determined by the Flory-Rehner equation. The cross-linked polymer showed much greater adhesive strength at room temperature, but the adhesive strength was quite low at higher temperature. The self-healing capability was studied by using scanning electron microscopy analysis. The thermal and dynamic mechanical properties of the thermally amendable cross-linked materials were investigated by thermogravimetric analysis and dynamic mechanical analysis.


Weather and shade effects on cow corral activities.
Visual scanning at 15-min intervals was used to monitor effects of shade forms and weather types on corral activities of cows. The method allowed one person to observe several dozen cows in a large pen, regardless of grouping pattern. Complete ration eating between 0600 and 1800 h was reduced 13% during hot weather, as compared to cool weather in unshaded corrals or pens with 4-m wide solid strip shades and unprotected manger . Hot afternoon fresh-feed eating response was reduced in comparison to cool weather 14% for unshaded corrals and 46% for strip shaded corrals having unprotected mangers . Cows having the strip shade plus a separate shade over the manger or a combined completely shaded free-stall rest and feeding area had double the ad libitum eating between fresh feed offerings during cool or warm weather over animals with unprotected mangers , and the response was tripled in hot weather. The percentage of cows resting or ruminating in the standing position increased linearly as temperatures increased. The effect was most evident for unshaded cows or those with the strip shade and unprotected manger . The percentage of cows drinking water or lingering around the trough without drinking increased in hot weather for unshaded animals. The observation method is taught easily, and results are useful in extension education programs.


Heterogeneous patterns of pH regulation in glial cells in the dorsal and ventral medulla.
We examined pH regulation in two chemosensitive areas of the brain, the retrotrapezoid nucleus (RTN) and the nucleus tractus solitarius (NTS), to identify the proton transporters involved in regulation of intracellular pH (pHi) in medullary glia. Transverse brain slices from young rats [postnatal day 8 (P8) to P20] were loaded with the pH-sensitive probe 2',7'-bis (2-carboxyethyl)-5,6-carboxyfluorescein after kainic acid treatment removed neurons. Cells were alkalinized when they were depolarized (extracellular K+ increased from 6.24 to 21.24 mM) in the RTN but not in the NTS. This alkaline shift was inhibited by 0.5 mM DIDS. Removal of CO2/HCO3- or Na+ from the perfusate acidified the glial cells, but the acidification after Na+ removal was greater in the RTN than in the NTS. Treatment of the slice with 5-(N-ethyl-N-isopropyl)amiloride (100 microM) in saline containing CO2/HCO3- acidified the cells in both nuclei, but the acidification was greater in the NTS. Restoration of extracellular Cl- after Cl- depletion during the control condition acidified the cells. Immunohistochemical studies of glial fibrillary acid protein demonstrated much denser staining in the RTN compared with the NTS. We conclude that there is evidence of Na+-HCO3- cotransport and Na+/H+ exchange in glia in the RTN and NTS, but the distribution of glia and the distribution of these pH-regulatory functions are not identical in the NTS and RTN. The differential strength of glial pH regulatory function in the RTN and NTS may also alter CO2 chemosensory neuronal function at these two chemosensitive sites in the brain stem.


Histologic malignancy grading of invasive ductal breast carcinoma. A regression analysis of prognostic factors in low-risk carcinomas from a multicenter trial.
In a prospective adjuvant trial including patients with primary operable breast cancer, invasive carcinomas of ductal type were subjected to histological malignancy grading. The parameters investigated were tubule formation, number of mitoses and cell pleomorphism. A Cox regression model for survival data was applied to evaluate the impact of the histological parameters on prognosis in 1809 patients with low-risk carcinomas. Cell pleomorphism proved superior to the other histologic tumor characteristics. It was found that low-risk invasive ductal carcinomas with severe cell pleomorphism had an excess recurrence intensity of 209% relative to carcinomas with no pleomorphism. It is therefore suggested that polymorphous invasive ductal breast carcinomas, other things being equal, should be regarded as high-risk tumors in future clinical trials. Finally it was found that the tripartite malignancy grading 1, 2 and 3 characterizing each of the histological parameters was not equidistant. Consequently, the traditional tripartite histologic scoring needs reconsideration.


First report of two cases of cryptococcosis in Tripoli, Libya, infected with Cryptococcus neoformans isolates present in the urban area.
Cryptococcosis is a potentially fatal fungal disease caused by the basidiomycetes yeasts Cryptococcus neoformans and C. gattii with high predilection to invade the central nervous system mainly in immunocompromised hosts. Skin can be secondarily involved in disseminated infection or be exceptionally involved as primary cutaneous infection by inoculation with contaminated materials. We report the first two Libyan cases of cryptococcal meningitis in HIV patients, in which one of them presented a secondary cutaneous involvement due to systemic dissemination. The first patient was a 17-year-old female, had fever, cough, headache and intractable vomiting as well as itchy water bumps on her skin and upper limbs. The cutaneous eruption prompted the accurate diagnosis. Cultures were positive for C. neoformans in both cerebrospinal fluid and skin specimens, as well as cryptococcal antigen was detected in serum. The isolate was identified, by molecular analysis, as C. neoformans AD-hybrid belonging to molecular type VNIII and mating type αAAα, the same genotype found for some environmental isolates recovered from olive trees in Tripoli. The second patient was a 36-years-old male with a long history of HIV on irregular treatment. Cryptococcal antigen in serum was positive and cultures yielded the growth of C. neoformans var. grubii, molecular type VNI and mating type αA. Both patients did not respond adequately to treatment and died of impaired central nervous system function and respiratory failure, respectively.


The molecular basis of electroporation.
Electroporation is a common method to introduce foreign molecules into cells, but its molecular basis is poorly understood. Here I investigate the mechanism of pore formation by direct molecular dynamics simulations of phospholipid bilayers of a size of 256 and of more than 2000 lipids as well as simulations of simpler interface systems with applied electric fields of different strengths. In a bilayer of 26 x 29 nm multiple pores form independently with sizes of up to 10 nm on a time scale of nanoseconds with an applied field of 0.5 V/nm. Pore formation is accompanied by curving of the bilayer. In smaller bilayers of ca. 6 x 6 nm, a single pore forms on a nanosecond time scale in lipid bilayers with applied fields of at least 0.4 V/nm, corresponding to transmembrane voltages of ca. 3 V. The presence of 1 M salt does not seem to change the mechanism. In an even simpler system, consisting of a 3 nm thick octane layer, pores also form, despite the fact that there are no charged headgroups and no salt in this system. In all cases pore formation begins with the formation of single-file like water defects penetrating into the bilayer or octane. The simulations suggest that pore formation is driven by local electric field gradients at the water/lipid interface. Water molecules move in these field gradients, which increases the probability of water defects penetrating into the bilayer interior. Such water defects cause a further increase in the local electric field, accelerating the process of pore formation. The likelihood of pore formation appears to be increased by local membrane defects involving lipid headgroups. Simulations with and without salt show little difference in the observed pore formation process. The resulting pores are hydrophilic, lined by phospholipid headgroups.


COVID-19 pandemic phases and female precocious puberty: The experience of the past 4 years (2019 through 2022) in an Italian tertiary center.
Since the outbreak of COVID-19 pandemic, several centers of pediatric endocrinology worldwide have observed a significant increase in the number of girls presenting with precocious or early puberty. We aimed to compare the incidence rates of female precocious puberty before and during the different phases of COVID-19 pandemic. We have retrospectively analyzed all the consultations recorded in the outpatient clinic database of the Endocrinology Unit of Bambino Gesù Children's Hospital, Rome, Italy, from the lockdown start in March 2020 up to September 2020, in comparison with the consultations recorded in the same months of 2019, 2021 and 2022. Age, height, weight, body mass index, Tanner's pubertal stage and bone age at presentation, birth weight, ethnicity, family history of central precocious puberty (CPP), maternal age at menarche, history of adoption were retrieved from clinical records. Serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH) both at baseline and after gonadotropin-releasing hormone (GnRH) stimulation, and basal estradiol levels were collected. In 2019, 78 girls with suspected precocious puberty were referred for endocrinological consultation, compared to 202 girls in 2020, 158 girls in 2021 and 112 girls in 2022. A significant increase in the proportion of girls diagnosed with rapidly progressive CPP was observed in 2020, compared to 2019 (86/202 vs. 18/78, p<0.01). In the following periods of 2021 and 2022, a gradual decrease in the number of cases of progressive CPP was evident, so much that the number of cases was not significantly different from that observed in 2019 (56/158 in 2021 and 35/112 in 2022, p=0.054 and p=0.216 respectively, compared to 2019). Our research suggests that drastic lifestyle changes, such as those imposed by COVID-19 lockdown, and the consequent stress may affect the regulation of pubertal timing. The remarkable increase in CPP cases observed during the 2020 first pandemic wave seems to be reduced in 2021 and 2022, concurrently with the progressive resumption of daily activities. These data seem to support the hypothesis of a direct relationship between profound life-style changes related to the pandemic and the rise in precocious puberty cases.


Diffusion Behavior of Chloride Ions in Concrete Box Girder under the Influence of Load and Carbonation.
In order to study the durability degradation characteristics of concrete box girder under load and carbonation and chloride ion erosion, a scale model of concrete box girder was made for experimental research. According to the test results, the diffusion characteristics of chloride ions in the concrete box girder under the coupling effect of load and carbon dioxide were analyzed. By revising the calculation formula of the existing chloride ion concentration considering multiple factors, a calculation model of chloride ion concentration considering the influence of carbonation was proposed, and the test results were verified. The results show that the chloride concentration of the box girder on the same cross section is non-uniformly distributed due to the shear lag effect and the spatial structure. After considering the effect of carbonation, the difference rate of the improved model proposed in this paper is generally within 10%. Compared with the original model, the difference rate is reduced by a maximum of 19%.


Knockout of Vasohibin-1 Gene in Mice Results in Healthy Longevity with Reduced Expression of Insulin Receptor, Insulin Receptor Substrate 1, and Insulin Receptor Substrate 2 in Their White Adipose Tissue.
Vasohibin-1 (Vash1), originally isolated as an endothelium-derived angiogenesis inhibitor, has a characteristic of promoting stress tolerance in endothelial cells (ECs). We therefore speculated that the lack of the vash1 gene would result in a short lifespan. However, to our surprise, vash1-/- mice lived significantly longer with a milder senescence phenotype than wild-type (WT) mice. We sought the cause of this healthy longevity and found that vash1-/- mice exhibited mild insulin resistance along with reduced expression of the insulin receptor (insr), insulin receptor substrate 1 (irs-1), and insulin receptor substrate 2 (irs-2) in their white adipose tissue (WAT) but not in their liver or skeletal muscle. The expression of vash1 dominated in the WAT among those 3 organs. Importantly, vash1-/- mice did not develop diabetes even when fed a high-fat diet. These results indicate that the expression of vash1 was required for the normal insulin sensitivity of the WAT and that the target molecules for this activity were insr, irs1, and irs2. The lack of vash1 caused mild insulin resistance without the outbreak of overt diabetes and might contribute to healthy longevity.


Intergroup contact and prejudice between Dutch majority and Muslim minority youth in the Netherlands.
This study deals with three relatively understudied issues in intergroup contact: negative contact, mediating mechanisms, and the minority perspective. Both direct and extended positive and negative contact experiences are included in the design. Intergroup anxiety is tested as a mediator between different forms of contact and prejudice, and status as Dutch majority or Muslim minority is used as a moderator. A sample of 317 Dutch majority (47.6% female) and 369 Muslim minority (52.0% female) youth, ranging in age from 12 to 19 years completed self-reports about contact experiences, intergroup threat, and prejudice. Results show that status as a Dutch majority or Muslim minority is a moderator in the relations between contact, intergroup anxiety, and prejudice. In the majority sample, all forms of direct and extended contact were related to prejudice and mediated by intergroup anxiety in the expected directions. In the Muslim minority sample, only positive contact was related to prejudice and mediated by intergroup anxiety in the expected direction. These findings underline that studies on intergroup relations should take both positive and negative contact experiences for intergroup attitudes into account as well as the majority or minority status of the groups involved. Moreover, the study suggests that partly different explanations may be needed for minority and majority groups for the role of intergroup contact in intergroup attitudes. (PsycINFO Database Record


Quantitative phase imaging for cell culture quality control.
The potential of quantitative phase imaging (QPI) with digital holographic microscopy (DHM) for quantification of cell culture quality was explored. Label-free QPI of detached single cells in suspension was performed by Michelson interferometer-based self-interference DHM. Two pancreatic tumor cell lines were chosen as cellular model and analyzed for refractive index, volume, and dry mass under varying culture conditions. Firstly, adequate cell numbers for reliable statistics were identified. Then, to characterize the performance and reproducibility of the method, we compared results from independently repeated measurements and quantified the cellular response to osmolality changes of the cell culture medium. Finally, it was demonstrated that the evaluation of QPI images allows the extraction of absolute cell parameters which are related to cell layer confluence states. In summary, the results show that QPI enables label-free imaging cytometry, which provides novel complementary integral biophysical data sets for sophisticated quantification of cell culture quality with minimized sample preparation. © 2017 International Society for Advancement of Cytometry.


Evaporation of ethanol and ethanol-water mixtures studied by time-resolved infrared spectroscopy.
The knowledge of the physics and the chemistry behind the evaporation of solvents is very important for the development of several technologies, especially in the fabrication of thin films from liquid phase and the organization of nanostructures by evaporation-induced self-assembly. Ethanol, in particular, is one of the most common solvents in sol-gel and evaporation-induced self-assembly processing of thin films, and a detailed understanding of its role during these processes is of fundamental importance. Rapid scan time-resolved infrared spectroscopy has been applied to study in situ the evaporation of ethanol and ethanol-water droplets on a ZnSe substrate. Whereas the evaporation rate of ethanol remains constant during the process, water is adsorbed by the ethanol droplet from the external environment and evaporates in three stages that are characterized by different evaporation rates. The adsorption and evaporation process of water in an ethanol droplet has been observed to follow a complex behavior: due to this reason, it has been analyzed by two-dimensional infrared correlation. Three different components in the water bending band have been resolved.


Apparent functionality but impractical quantification of neurotensin receptors on human peripheral lymphocytes.
Neurotensin (NT), a neuromodulator, is also thought to play an immunomodulatory role. We sought to confirm the effects of NT on proliferation in human peripheral blood lymphocytes (PBLs; n = 10), to characterize the binding properties of the NT receptor using Scatchard analysis, and to measure NT receptors using blood volumes (50 ml) applicable to clinical investigation. Incubation of phytohemagglutinin-stimulated PBLs with NT (10[-10] to 10[-13] M) in the presence of 0.07% normal human serum (72 h at 37 degrees C) significantly enhanced proliferation (p < 0.001), which is consistent with PBLs possessing functional NT receptors. However, the various experimental conditions tested constantly yielded low specific NT binding to human PBLs, and hence Scatchard analysis was impossible for 50-ml blood samples. Our data confirm the existence of a link between NT and the immune system and support a physiological significance for this link. However, measurement of NT receptor binding in readily available cells proved unsuitable for clinical investigation.


[A randomized controlled pilot on chronic non-specific low back pain treated with the superficial needling therapy combined with mild moxibustion].
To compare the differences in the clinical therapeutic effects on chronic non-specific low back pain (CNLBP) between the combined treatment of the superficial needling technique and mild moxibustion and the traditional warm acupuncture. A total of 60 patients were randomized into a combined treatment group and a warm acupuncture group, 30 cases in each one. In the combined treatment group, the superficial needling technique was used in combination with the mild moxibustion with moxa box at the muscle region of the bladder meridian on the back. In the warm acupuncture group, the traditional warm acupuncture was adopted on the lumbar region and the upper back. Each treatment lasted 30 min, 3 times a week. The treatment was provided continuously for 3 weeks. The follow-up visit lasted 3 months. Separately, before, 3 times after treatment and at the end of treatment, as well as after the follow-up visit, the visual analogue scale (VAS), the range of motion of the lumbar region (ROM) and the Oswestry disability index (ODI) were observed in the patients of the two groups. During and at the end of treatment, as well as at the follow-up visit, VAS score, ROM and ODI were all improved as compared with those before treatment in the two groups (all P<0.05). The results in the combined treatment group were better than those in the warm acupuncture (all P<0.05). At the end of treatment and the follow-up stage, VAS score, ROM and ODI were better than those during treatment in the two groups (all P<0.05). At the follow-up stage, VAS score, ROM of the backward extension and rotation of spine, as well as ODI were better than those at the end of treatment in the two groups (all P<0.05). The combined treatment of the superficial needling technique and mild moxibustion relieves pain, improves the ROM of the lumbar region and reduces the functional disability in CNLBP. This combined therapy achieves the better effects as compared with the traditional warm acupuncture and is high in the patient's compliance.


Surgical treatment of recidivist lymphedema.
Lymphedema is a chronic disease with a progressively ingravescent evolvement and an appearance of recurrent complications of acute lymphangitic type; in nature it is mostly erysipeloid and responsible for a further rapid increase in the volume and consistency of edema. The purpose of this work is to present our experience in the minimally invasive treatment for recurrence of lymphedema; adapting techniques performed in the past which included large fasciotomy with devastating results cosmetically; but these techniques have been proposed again by the use of endoscopic equipment borrowed from the advanced laparoscopy surgery, which allows a monoskin access of about one cm.


Power Doppler ultrasonographic assessment of the ankle in patients with inflammatory rheumatic diseases.
Ankle involvement is frequent in patients with inflammatory rheumatic diseases, but accurate evaluation by physical examination is often difficult because of the complex anatomical structures of the ankle. Over the last decade, ultrasound (US) has become a practical imaging tool for the assessment of articular and periarticular pathologies, including joint synovitis, tenosynovitis, and enthesitis in rheumatic diseases. Progress in power Doppler (PD) technology has enabled evaluation of the strength of ongoing inflammation. PDUS is very useful for identifying the location and kind of pathologies in rheumatic ankles as well as for distinguishing between inflammatory processes and degenerative changes or between active inflammation and residual damage. The aim of this paper is to illustrate the US assessment of ankle lesions in patients with inflammatory rheumatic diseases, including rheumatoid arthritis, spondyloarthritis, and systemic lupus erythematosus, focusing on the utility of PDUS.


Charge-Transfer Emission in Organoboron-Based Biphenyls: Effect of Substitution Position and Conformation.
A series of organoboron-based biphenyls o,o'-NMe2, o,p'-NMe2, p,p'-NMe2, which contain an electron-donating NMe2 and an electron-accepting BMes2 groups at o,o'-, o,p'-, p,p'-positions of biphenyl skeleton, respectively, as well as o,o'-NBn2, which contains more bulky NBn2 rather than NMe2, were fully characterized to explore the effect of structural modification on the intramolecular charge-transfer emissions. In addition to significant effect of substitution position on photophysical properties, remarkable influence of conformation was also observed for o,o'-substituted compounds. The emission is substantially blue-shifted as conformation changes from the location of NMe2 and BMes2 at same side of biphenyl axis with a close B···N distance, and thus direct B···N electronic interaction in o,o'-NMe2, to the location of NBn2 and BMes2 on two opposite sides in o,o'-NBn2. And o,o'-NMe2 exhibits the longest emission wavelength, but the shortest absorption wavelength, and thus largest Stokes shift among these four organoboron-based biphenyls in both solution and solid state. The theoretical calculations demonstrated that the unique structure of o,o'-NMe2, in which boryl and amino located at the same side of biphenyl axis with close B···N distance and direct B···N electronic interaction, is helpful to stabilize the lowest singly occupied orbital in the exited state.


Ocimum basilicum Essential Oil Modulates Hematotoxicity, Oxidative Stress, DNA Damage, and Cell Cycle Arrest Induced by β-cyfluthrin in Rat Liver.
Pesticides are used in large quantities infrequently, resulting in environmental damage and health issues. The goal of the current study was to explore the ameliorating effect of Ocimum basilicum (Basil) leaves essential oil versus the harmful effects of β-cyfluthrin in rat liver. Male Wistar rats were classified at random into four groups; negative control (corn oil), basil leaves essential oil (BEO, 3 ml/kg), β-cyfluthrin (positive control) (β-Cyf; 15 mg/kg BW, 1/25 LD50), and BEO plus β-Cyf, respectively. The rats were given their doses orally every day for a month. Results revealed that BEO yielded 6.32 mg/g with 33 identified components, representing 97% of the total oil. BEO implicated a considerable level of total phenolic contents, DPPH radical scavenging capacity, ABTS activity, and FRAP. The treatment of β-Cyf dramatically elevated lipid peroxidation (TBARS and H2O2) (LPO), protein oxidation (PC, AOPP, and HYP), and considerably reduced enzymatic (SOD, CAT, GPx, GR, and GST) and non-enzymatic (GSH) antioxidants. After β-Cyf treatment, hematological parameters, body and liver weights, enzyme activity (AST, ALT, ALP, and LDH), as well as protein, albumin, globulin, and total bilirubin levels were all considerably affected. Furthermore, β-Cyf increased the expression of pro-inflammatory genes (TNF-α, IL-6) as well as DNA damage and cell cycle arrest in the G0/G1 phase and decreased the number of cells in S and G2/M phase of liver cells. Moreover, rats given BEO then intoxicated with β-Cyf showed substantial changes in the majority of the parameters tested. Finally, BEO was shown to have high antioxidant efficacy in combating β-Cyf toxicity because of its high phenolic content.


Off-pump surgery: is it beneficial in patients with left ventricular dysfunction?
On-pump coronary artery bypass surgery remains the gold standard for complex multivessel disease. Off-pump revascularization has matured as a technique over the last twenty years, but is used in less than 20% of cases worldwide. The poor uptake has been attributed to the significant learning curve in learning the procedure and conflicting evidence reports, together with concerns over mortality related conversion, graft patency and completeness of revascularization. Given these concerns, patient selection continues to be paramount and the subgroups that benefit most are hotly debated. Patients with left ventricular dysfunction constitute a high-risk subgroup which is enlarging in size. There is some evidence to suggest that avoidance of cardiopulmonary bypass in this group may lead to superior results in terms of early mortality, non-cardiac complications and organ dysfunction. Even with the theoretical risk of incomplete revascularization, the technique may be an attractive option in managing high-risk patients.


Decision-theoretic designs for dose-finding clinical trials with multiple outcomes.
A decision-theoretic framework is proposed for designing sequential dose-finding trials with multiple outcomes. The optimal strategy is solvable theoretically via backward induction. However, for dose-finding studies involving k doses, the computational complexity is the same as the bandit problem with k-dependent arms, which is computationally prohibitive. We therefore provide two computationally compromised strategies, which is of practical interest as the computational complexity is greatly reduced: one is closely related to the continual reassessment method (CRM), and the other improves CRM and approximates to the optimal strategy better. In particular, we present the framework for phase I/II trials with multiple outcomes. Applications to a pediatric HIV trial and a cancer chemotherapy trial are given to illustrate the proposed approach. Simulation results for the two trials show that the computationally compromised strategy can perform well and appear to be ethical for allocating patients. The proposed framework can provide better approximation to the optimal strategy if more extensive computing is available.


Pyrexia, anorexia, adipsia, and depressed motor activity in rats during systemic inflammation induced by the Toll-like receptors-2 and -6 agonists MALP-2 and FSL-1.
Macrophage-activating lipopeptide-2 (MALP-2) from Mycoplasma fermentans has been identified as a pathogen-associated molecular pattern of Mycoplasmas that causes activation of the innate immune system through the activation of the heterodimeric Toll-like receptors (TLRs)-2 and -6. The aim of this study was to characterize the ability of MALP-2 and a synthetic analog fibroblast-stimulating lipopeptide-1 (FSL-1; represents the NH2-terminal sequence of a lipoprotein from M. salivarium) to act as exogenous pyrogens, to induce formation of cytokines (endogenous pyrogens), and to cause sickness behavior, such as depressed motor activity, anorexia, and adipsia. For this purpose, body temperature, activity, food intake, and water intake were recorded for 3 days by use of telemetry devices in several groups of rats treated with MALP-2/FSL-1 or the respective control solutions. Intraperitoneal injections of FSL-1 caused fever at doses of 10 or 100 microg/kg, which was preceded by a pronounced phase of hypothermia in response to a dose of 1,000 microg/kg. The maximal fever (a peak of 1.5 degrees C above baseline) was caused by the 100 microg/kg dose with almost identical responses to both MALP-2 and FSL-1. Fever was accompanied by pronounced rises of the proinflammatory cytokines TNF and IL-6 in plasma. Treatment with the TLR-2 and -6 agonists further induced a dose-dependent manifestation of anorexia and adipsia, as well as a reduction of motor activity. We could thus demonstrate that activation of TLR-2 and -6 can induce systemic inflammation in rats accompanied by the classical signs of brain-controlled illness responses.


[Identification of LicC activity in pneumococcal infection].
To clone and express licC and its truncated form genes (25 amino acids from 3'-terminus were deleted and named deltaLicC) of Streptococcus pneumoniae, analyze the enzymatic activities of the proteins. The recombinant plasmid pQE80-licC, pQE80-deltalicC were constructed, and the target proteins were expressed in E. coli BL21 under isopropy-beta-D-thiogalactoside (IPTG) induction. The proteins' activities were determined using bioluminescence test based on firefly luciferase assay system. The prokaryotic expression vector pQE80-licC, pQE80-deltalicC were successfully constructed and identified.The soluble proteins were obtained through inducing expression in E. coli BL21. It was showed that the activity of deltaLicC was markedly lower than that of LicC (P<0.05). The homemade bioluminescence assay method can miner the activity of LicC reliably and accurately. The important role of 25 amino acids from 3'-terminus for the activity of LicC was confirmed, and it was suggested that suppressing of LicC maybe was a useful method for treatment S.pneumococcal infection, especially drug resistant strain.


Structure modification and extracellular polymeric substances conversion during sewage sludge biodrying process.
Biodrying, an economical and energy-saving biomass waste treatment, removes water from waste using the biological heat generated by organic matter degradation. Technical limitations associated with dewatering complicate the biodrying of sewage sludge. This study investigated the sludge alteration associated with its water removal, focusing on sludge form, extracellular polymeric substances, and free water release. An auto-feedback control technology was used for the biodrying; a scanning electron microscope was used to record the morphological change; three-dimensional excitation-emission matrix fluorescence spectroscopy was used to analyze extracellular polymeric substances (EPS) variation, and time domain reflectometry was used to assess the free water release. Over the 20-day biodrying, there was a 62% water removal rate during the first thermophilic phase. Biodrying created a hollow and stratified sludge structure. Aromatic proteins and soluble microbial byproducts in the EPS were significantly degraded. The thermophilic phase was the phase resulting in the greatest free water release.


The Belisana spiders (Araneae: Pholcidae) from Xishuangbanna Tropical Botanical Garden, Yunnan, China.
Five new species of the spider genus Belisana Thorell, 1898 are described based on material collected in the Xishuangbanna Tropical Botanical Garden (Yunnan, China). They are: B. dian Yao Li sp. nov. (male), B. gupian Yao Li sp. nov. (male, female), B. chenjini Yao Li sp. nov. (male, female), B. xtbg Yao Li sp. nov. (male, female), and B. yangxiaodongi Yao Li sp. nov. (male, female). B. schwendingeri Huber, 2005 and B. zhengi Yao, Pham Li, 2015 are reported from China for the first time. All material studied is deposited in the Institute of Zoology, Chinese Academy of Sciences (IZCAS) in Beijing, China.


Metagenomic characterization of Chesapeake Bay virioplankton.
Viruses are ubiquitous and abundant throughout the biosphere. In marine systems, virus-mediated processes can have significant impacts on microbial diversity and on global biogeocehmical cycling. However, viral genetic diversity remains poorly characterized. To address this shortcoming, a metagenomic library was constructed from Chesapeake Bay virioplankton. The resulting sequences constitute the largest collection of long-read double-stranded DNA (dsDNA) viral metagenome data reported to date. BLAST homology comparisons showed that Chesapeake Bay virioplankton contained a high proportion of unknown (homologous only to environmental sequences) and novel (no significant homolog) sequences. This analysis suggests that dsDNA viruses are likely one of the largest reservoirs of unknown genetic diversity in the biosphere. The taxonomic origin of BLAST homologs to viral library sequences agreed well with reported abundances of cooccurring bacterial subphyla within the estuary and indicated that cyanophages were abundant. However, the low proportion of Siphophage homologs contradicts a previous assertion that this family comprises most bacteriophage diversity. Identification and analyses of cyanobacterial homologs of the psbA gene illustrated the value of metagenomic studies of virioplankton. The phylogeny of inferred PsbA protein sequences suggested that Chesapeake Bay cyanophage strains are endemic in that environment. The ratio of psbA homologous sequences to total cyanophage sequences in the metagenome indicated that the psbA gene may be nearly universal in Chesapeake Bay cyanophage genomes. Furthermore, the low frequency of psbD homologs in the library supports the prediction that Chesapeake Bay cyanophage populations are dominated by Podoviridae.


RNA purification from epidermal suction blisters.
Certain inflammatory skin diseases are accompanied by increased cytokine concentrations in the epidermis. To determine whether these cytokines are synthesized in the epidermis or exported from underlying tissues, epidermal RNA was analysed for the presence of their messenger RNAs. We report a method for RNA extraction from pure epidermal samples isolated by the suction blister method. The yield of total RNA was sufficient for hybridization experiments (12-38 micrograms per seven blisters, 5 mm in diameter). Using RNA extracted by this method, we have demonstrated the presence of messenger RNA for glyceraldehyde-3-phosphate-dehydrogenase in 13 preparations from suction blisters obtained from tuberculin skin reactions, positive patch test reactions, or normal skin. We did not, however, observe messenger RNA for interleukin 1 alpha or 8 in these preparations.


The Rcs phosphorelay: a complex signal transduction system.
RcsC, RcsB, and RcsA were first identified as a sensor kinase, a response regulator, and an auxiliary regulatory protein, respectively, regulating the genes of capsular polysaccharide synthesis. Recent advances have demonstrated that these proteins are part of a complex phosphorelay, in which phosphate travels from the histidine kinase domain in RcsC to a response regulator domain in the same protein; from there to a phosphotransfer protein, RcsD; and from there to RcsB. In addition to capsule synthesis, which requires the unstable regulatory protein RcsA, RcsB also stimulates transcription of a small RNA, RprA; the cell division gene ftsZ; and genes encoding membrane and periplasmic proteins, including the osmotically inducible genes osmB and osmC. The Rcs system appears to play an important role in the later stages of biofilm development; induction of Rcs signaling by surfaces is consistent with this role.


Enzyme-Powered Hollow Mesoporous Janus Nanomotors.
The development of synthetic nanomotors for technological applications in particular for life science and nanomedicine is a key focus of current basic research. However, it has been challenging to make active nanosystems based on biocompatible materials consuming nontoxic fuels for providing self-propulsion. Here, we fabricate self-propelled Janus nanomotors based on hollow mesoporous silica nanoparticles (HMSNPs), which are powered by biocatalytic reactions of three different enzymes: catalase, urease, and glucose oxidase (GOx). The active motion is characterized by a mean-square displacement (MSD) analysis of optical video recordings and confirmed by dynamic light scattering (DLS) measurements. We found that the apparent diffusion coefficient was enhanced by up to 83%. In addition, using optical tweezers, we directly measured a holding force of 64 ± 16 fN, which was necessary to counteract the effective self-propulsion force generated by a single nanomotor. The successful demonstration of biocompatible enzyme-powered active nanomotors using biologically benign fuels has a great potential for future biomedical applications.


Kaposi's sarcoma associated with multiple myeloma.
I have described a patient with multiple myeloma in whom Kaposi's sarcoma of the skin developed during treatment with an alkylating agent. The association of a plasma cell dyscrasia, such as multiple myeloma, and Kaposi's sarcoma is uncommon. The possible role of angiogenesis in deranged immunologic responses is addressed.


Mechanistic insights into the catalytic asymmetric allylboration of ketones: Brønsted or Lewis acid activation?
Binaphthol ligands promote the enantioselective addition of allylboronates to ketones. In this study, we use DFT calculations to establish the identity of the reacting chiral species. Our results show that a cyclic Lewis acid-activated boronate is the most reactive species on the basis of calculated energy barriers, and it is only this species that leads to the correct enantiomer. The stereoinduction can be rationalized in terms of the competing chairlike transition structures.


Discovering Microcircuit Secrets With Multi-Spot Imaging and Electrophysiological Recordings: The Example of Cerebellar Network Dynamics.
The cerebellar cortex microcircuit is characterized by a highly ordered neuronal architecture having a relatively simple and stereotyped connectivity pattern. For a long time, this structural simplicity has incorrectly led to the idea that anatomical considerations would be sufficient to understand the dynamics of the underlying circuitry. However, recent experimental evidence indicates that cerebellar operations are much more complex than solely predicted by anatomy, due to the crucial role played by neuronal and synaptic properties. To be able to explore neuronal and microcircuit dynamics, advanced imaging, electrophysiological techniques and computational models have been combined, allowing us to investigate neuronal ensembles activity and to connect microscale to mesoscale phenomena. Here, we review what is known about cerebellar network organization, neural dynamics and synaptic plasticity and point out what is still missing and would require experimental assessments. We consider the available experimental techniques that allow a comprehensive assessment of circuit dynamics, including voltage and calcium imaging and extracellular electrophysiological recordings with multi-electrode arrays (MEAs). These techniques are proving essential to investigate the spatiotemporal pattern of activity and plasticity in the cerebellar network, providing new clues on how circuit dynamics contribute to motor control and higher cognitive functions.


Three-dimensional endoanal sonography in assessing anal canal injury.
Instrument design limits endosonography of the anal canal to the axial plane, with no capability for longitudinal imaging or measurement. Using three-dimensional reconstructions, the relationship between the radial and linear extent of an anal sphincter tear has been explored, and sex differences in anal canal and sphincter length have been established. Three-dimensional reconstructions were performed in 20 controls and 24 patients with faecal incontinence found to have 25 external and five internal sphincter defects. The radial and linear extent of any sphincter tear was measured. In controls the length of the sphincters was compared with the total anal canal length, and the maximum and mean internal sphincter thickness was compared. The radial angle of an internal or external sphincter defect was significantly related to its length (R2 = 96.8 per cent and R2 = 84.4 per cent respectively; both P < 0.001). The anal canal was longer in men than in women (mean(s.d.) 32.6(5.3) versus 25.1(3.4) mm; P < 0.001). The internal anal sphincter was also longer in men (25.6(6.3) versus 19.8(4.0) mm; P < 0.02), but the mean internal sphincter length as a percentage of total anal canal length did not differ (78.3 versus 78.7 per cent; P not significant). The anterior external anal sphincter was longer in men than in women (32.6(5.3) versus 15.3(2.8) mm; P < 0.001), and formed a greater percentage of total anal canal length (100 versus 62.9 per cent; P < 0.001). Multiplanar imaging has revealed a direct relationship between the length of a sphincter tear and its radial extent as shown on axial scanning. Marked sex differences in sphincter configuration have been demonstrated. In women the shorter anterior sphincter length highlights the risk of complete sphincter disruption with extensive tears.


Comparison of temporomandibular disorders in Iranian dental and nondental students.
The aim of this study was to define the prevalence and severity of temporomandibular disorders (TMD) among Iranian dental and nondental students who attended Shiraz University of Medical Sciences. The samples consisted of 200 subjects, 100 dental and 100 nondental students (equal distribution between males and females). Subjects ages ranged from 18 to 30 years (24.07 ± 2.93). A functional evaluation was performed using the Helkimo dysfunction index (Di). Data were evaluated by the Chi-square test. Among the total study population, 71% showed some degree of dysfunction. Prevalence ratio in dental students was 80%, while in nondental students was 62% (p > 0.05, odds ratio = 0.679). With respect to gender, women (80%) were more affected than men (62%). A significant relationship was found between gender and the occurrence of TMD (p > 0.05). A high prevalence of signs of TMD in Iranian university students was seen which was greater in dental students and women. The results of this study showed that the prevalence of TMD among dental students and women was significantly higher than nondental students and men. These findings might be due to poor postural style or emotional stress during dental practice.


Oleic acid enhances G protein coupled receptor 43 expression in bovine intramuscular adipocytes but not in subcutaneous adipocytes.
We hypothesized that fatty acids would differentially affect G protein coupled receptor (GPR) 43 mRNA expression and GPR43 protein concentrations in bovine intramuscular (IM) and subcutaneous (SC) adipocytes. The GPR43 protein was detected in bovine liver, pancreas, and semimembranosus (MUS) muscle in samples taken at slaughter. Similarly, GPR43 protein levels were similar in IM adipose tissue and SM muscle but was barely detectable in SC adipose tissue. Primary cultures of IM and SC stromal vascular cells were isolated from bovine adipose tissues. Oleic acid (100 μ) stimulated PPARγ gene expression and decreased stearoyl-CoA desaturase (SCD) gene expression but had no effect on GPR43 gene expression, which was readily detectable in both IM and SC adipocytes. Differentiation cocktail (Diff; 10 μ insulin, 4 μ dexamethasone, and 10 μ ciglitizone) stimulated CCAAT/enhancer-binding protein β (C/EBPβ) and PPARγ gene expression in SC but not IM adipocytes, but Diff increased SCD gene expression in both cell types. Linoleic acid (10 µ) increased PPARγ gene expression relative to Diff cocktail in SC adipocytes, whereas linoleic acid and α-linolenic decreased SCD gene expression relative to control adipocytes and adipocytes incubated with Diff ( < 0.05). Increasing concentrations of oleic acid (1, 10, 100, and 500 μM) increased GPR43 protein and mRNA expression in IM but not SC adipocytes. These data indicated that oleic acid alters mRNA and protein concentrations of GPR43 in bovine IM adipocytes.


Chronotropic effects of nitric oxide in the denervated human heart.
Nitric oxide synthase is expressed in the sino-atrial node and animal data suggests a direct role for nitric oxide on pacemaker activity. Study of this mechanism in intact humans is complicated by both reflex and direct effects of nitric oxide on cardiac autonomic control. Thus, we have studied the direct effects of nitric oxide on heart rate in human cardiac transplant recipients who possess a denervated donor heart. In nine patients, the chronotropic effects of systemic injection of the nitric oxide synthase inhibitor N(G)-monomethyl-L-arginine (L-NMMA) (3 mg kg(-1)) or increasing bolus doses of the nitric oxide donor, sodium nitroprusside (SNP), were studied. Injection of L-NMMA increased mean arterial pressure by 17 +/- 2 mmHg (mean +/- S.E.M.; P < 0.001) and also had a significant negative chronotropic effect, lengthening the R-R interval by 54 +/- 8 ms (P < 0.001). This bradycardia was not reflex in origin since injection of the non-NO-dependent vasoconstrictor, phenylephrine (100 microg) achieved a similar rise in mean arterial pressure (18 +/- 3 mmHg; P < 0.001) but failed to change R-R interval duration (Delta R-R = -3 +/- 4 ms). Furthermore, no change in levels of circulating adrenaline was observed with L-NMMA. Conversely, injection of sodium nitroprusside resulted in a positive chronotropic effect with a dose-dependent shortening of R-R interval duration, peak Delta R-R = -25 +/- 8 ms with 130 microg (P < 0.01). These findings indicate that nitric oxide exerts a tonic, direct, positive chronotropic influence on the denervated human heart. This is consistent with the results of animal experiments showing that nitric oxide exerts a facilitatory influence on pacemaking currents in the sino-atrial node.


Image detection and enhancement.
An optimal filtering approach is taken to the problem of detecting and enhancing a photographic image immersed in background or noise. The minimum mean square error criterion is shown to be not relevant to the optical employment of the Wiener filter. It is shown theoretically and demonstrated by simulation that, even without information about the background, a knowledge of the target spectrum alone makes it possible to find and display such a target without distortion while eliminating all uncorrelated background, despite the fact that target and background spectra actually overlap, and virtually without regard to noise level.


Hemodynamic changes due to infiltration of the scalp with epinephrine-containing lidocaine solution: a hypotensive episode before craniotomy.
Epinephrine-containing lidocaine solution is commonly infiltrated on the scalp before craniotomy. But the hemodynamic changes caused by epinephrine-containing lidocaine solution have been less intensely studied. A prospective randomized double blind control study was designed to observe hemodynamic changes caused by epinephrine-containing lidocaine solution in neurosurgical operations under general anesthesia. One hundred twenty patients undergoing scheduled craniotomy were allocated randomly to 4 groups. All the patients received 1% lidocaine 16 mL with different dose (concentration) epinephrine: group 1 with 40 microg (2.5 microg/mL); group 2 with 80 microg (5 microg/mL); group 3 with 160 microg (10 microg/mL); and group 4 (control group) without epinephrine. mean arterial pressure (MAP) and heart rate were recorded at 30-second interval in 5 minutes after the beginning of local infiltration. In group 1, group 2, and group 3, the lowest MAP and the highest MAP during this period also were recorded. Bleeding was assessed after raising the craniotomy flap. Compared with the baseline, significant hemodynamic changes, particularly decrease in MAP with increase in heart rate at 1.5 minutes after the beginning of local infiltration, were observed in group 1, group 2, and group 3 (P<0.001), but not in group 4. The highest MAP increased significantly compared with the baseline in group 3 (P<0.05), but not in group 1 or group 2. Epinephrine-containing lidocaine solution reduced bleeding significantly (P<0.01). Infiltration with epinephrine-containing lidocaine solution elicits temporary but significant hemodynamic changes including hypotension before craniotomy. Commonly clinically used concentrations of epinephrine (2.5 to 10 microg/mL) can reduce the bleeding on the scalp.


A synthesis of the literature on trauma-informed care.
Patients with a history of traumatic life events can become distressed or re-traumatized as the result of healthcare experiences. These patients can benefit from trauma-informed care that is sensitive to their unique needs. However, despite the widespread prevalence of traumatic life experiences such as sexual assault and intimate partner violence, trauma-informed care has not been widely researched or implemented. The purpose of this synthesis of the literature is to examine existing research on trauma-informed care for survivors of physical and sexual abuse. The following themes are discussed: trauma screening and patient disclosure, provider-patient relationships, minimizing distress and maximizing autonomy, multidisciplinary collaboration and referrals, and trauma-informed care in diverse settings. This synthesis also explores implications for trauma-informed care research, practice and policy. The themes identified here could be used as a framework for creating provider and survivor educational interventions and for implementing trauma-informed care across disciplines. The findings of this synthesis support further research on patient and provider experiences of trauma-informed care, and research to test the efficacy of trauma-informed care interventions across healthcare settings. Universal implementation of trauma-informed care can ensure that the unique needs of trauma survivors as patients are met, and mitigate barriers to care and health disparities experienced by this vulnerable population.


Release dependence to a paired stimulus at a synaptic release site with a small variable pool of immediately releasable vesicles.
Monte Carlo simulations were performed on a release model based on experimental data from single glutamatergic synapses containing a single release site in the hippocampal CA1 region of the neonatal rat. These simulations explored what can be learned about the release process by examining how the release probability in response to the second stimulus (P(2)) of a paired stimulus to a synapse depends on the release in response to the first stimulus. Comparisons between experimental data from a number of individual synapses and the simulated data support the notion that the immediately releasable vesicle pool is small (approximately one) and shows substantial intertrial variation. The simulations also show that the release dependence of P(2) is not necessarily an indicator of either intertrial variation in Ca(2+) influx, feedback effects of released transmitter, or activation failure of the axon.


Thapsigargin, Origin, Chemistry, Structure-Activity Relationships and Prodrug Development.
Thapsigargin was originally isolated from the roots of the Mediterranean umbelliferous plant Thapsia garganica in order to characterize the skin irritant principle. Characteristic chemical properties and semi-syntheses are reviewed. The biological activity was related to the subnanomolar affinity for the sarco/endoplasmic reticulum calcium ATPase. Prolonged inhibition of the pump afforded collapse of the calcium homeostasis and eventually apoptosis. Structure-activity relationships enabled design of an equipotent analogue containing a linker. Conjugation of the analogue containing the linker with peptides, which only are substrates for either prostate specific antigen (PSA) or prostate specific membrane antigen (PSMA) enabled design of prodrugs targeting a number of cancer diseases including prostate cancer (G115) and hepatocellular carcinoma (G202). Prodrug G202 has under the name of mipsagargin in phase II clinical trials shown promising properties against hepatocellular carcinoma.


Evaluation of the contribution of cyclooxygenase 1 and cyclooxygenase 2 to the production of PGE2 and PGF2 alpha in epithelial cells from bovine endometrium.
In ruminants, the production of prostaglandins by the endometrium is critical for recognition of pregnancy. In the absence of an embryonic signal, luteolytic pulses of PGF(2 alpha) are released by the uterus. In contrast, the presence of a viable conceptus reduces the production of PGF(2 alpha) relative to PGE(2) and prevents luteolysis through the release of trophoblastic interferon (IFN-tau). Initially, it was thought that epithelial and stromal endometrial cells were specialized in the production of a single type of prostaglandin. However, purified cell populations of both types of cell can produce PGF(2 alpha) and PGE(2); therefore, selective production of PGF(2 alpha) and PGE(2) must be regulated within each type of cell. Two distinct prostaglandin synthases, cyclooxygenase 1 and cyclooxygenase 2, are involved in prostaglandin production and each may catalyse the production of a different prostaglandin. This possibility was investigated in cultured epithelial cells from bovine endometrium. Cells were treated with oxytocin or arachidonic acid, and expression of cyclooxygenase 1 and cyclooxygenase 2 proteins was monitored over time and correlated with prostaglandin accumulation. Cells were also treated with increasing doses of inhibitors of cyclooxygenase 1 or cyclooxygenase 2 (non-steroidal anti-inflammatory drugs; NSAIDs) with or without arachidonic acid or oxytocin: flurbiprofen (0-50 micromol l(-1)) was used as a non-selective inhibitor; valeryl salicylate (0-500 micromol l(-1)) was used as a cyclooxygenase 1 inhibitor and NS-398 (0-1 micromol l(-1)) was used as a cyclooxygenase 2 inhibitor. After stimulation with arachidonic acid or oxytocin, prostaglandin production and expression of cyclooxygenase 2 protein were increased. All inhibitors were able to block basal and stimulated prostaglandin production. These results indicate that in endometrium most, if not all, prostaglandin production is probably processed through cyclooxygenase 2.


Implementation of a stroke code system in Mobile, Alabama. Diagnostic and therapeutic yield.
There is now therapy of proven benefit for acute ischemic stroke. Successful interventional therapy for stroke patients requires implementation of a system that facilitates rapid triage and diagnostic evaluation. We initiated a 24-hour, 7-day-per-week stroke code system at the University of South Alabama Hospitals and prospectively collected data from the first 100 patients whose clinical presentations triggered this system. Seventy-eight patients (78%) had acute ischemic stroke. Of the remaining 22, 9 had evidence of intracerebral hemorrhage. The most common nonstroke diagnosis was seizure (n = 5). Forty-eight of the 87 stroke patients (55%) presented within 6 hours of stroke onset (40/78 = 51% of the ischemic stroke patients), and 35 of the 87 (40%) presented within 3 hours of onset (28/78 = 36% of the ischemic stroke patients). Thirty-one (31% of the group overall; 40% of the ischemic stroke patients) were eligible for acute therapy. Twenty-five of these eligible patients were entered into a treatment study, 4 declined participation, and 2 were treated with open-label tissue plasminogen activator. Implementation of a stroke code system may result in a high yield of patients with acute stroke and relatively few "stroke mimickers." A significant proportion of all cases generated will be eligible for acute treatment under current experimental protocols or with tissue plasminogen activator, but the majority will not.


Theoretical study of plutonium(IV) complexes formed within the PUREX process: a proposal of a plutonium surrogate in fire conditions.
We present a relativistic quantum chemical study to determine the best surrogate for plutonium(IV) to be used in experimental investigations of the behavior of plutonium-nitrate-TBP in fire conditions that might occur in the nuclear fuel refining process known as PUREX. In this study geometries and stabilities of Pu(NO3)6(2-) and Pu(NO3)4(TBP)2 complexes were compared to that of equivalent complexes of selected elements from the lanthanide and actinide series (Ce, Th, U) chosen on the basis of similar ionic radii and stability as tetravalent species. PBE and PBE0 DFT functionals have proven to be sufficient and affordable for qualitative studies, performing as good as the wave function based correlated method MP2. On the basis of our results, cerium(IV) appears to be a good surrogate for plutonium(IV).


A cluster-based approach to selecting representative stimuli from the International Affective Picture System (IAPS) database.
The International Affective Picture System (IAPS; Lang, Bradley, & Cuthbert, 2008) is a stimulus database that is frequently used to investigate various aspects of emotional processing. Despite its extensive use, selecting IAPS stimuli for a research project is not usually done according to an established strategy, but rather is tailored to individual studies. Here we propose a standard, replicable method for stimulus selection based on cluster analysis, which re-creates the group structure that is most likely to have produced the valence arousal, and dominance norms associated with the IAPS images. Our method includes screening the database for outliers, identifying a suitable clustering solution, and then extracting the desired number of stimuli on the basis of their level of certainty of belonging to the cluster they were assigned to. Our method preserves statistical power in studies by maximizing the likelihood that the stimuli belong to the cluster structure fitted to them, and by filtering stimuli according to their certainty of cluster membership. In addition, although our cluster-based method is illustrated using the IAPS, it can be extended to other stimulus databases.


Autocrine regulation of macrophage differentiation and 92-kDa gelatinase production by tumor necrosis factor-alpha via alpha5 beta1 integrin in HL-60 cells.
Tumor necrosis factor-alpha (TNF-alpha) gene is one of the early response genes induced by phorbol 12-myristate 13-acetate (PMA) in human HL-60 myeloid leukemia cells. In the present study, we examined the role of the TNF-alpha autocrine loop in PMA-induced macrophage differentiation and gene expression of 92- and 72-kDa gelatinases (MMP-9 and MMP-2). In HL-60 cells, PMA inhibited cell proliferation and induced cell adhesion and spreading, expression of surface maturation marker OKM1 and phagocytic activity, as well as the expression of both gelatinases, which all characterize the macrophage phenotype. In contrast, TNF-alpha alone was only effective in inhibiting cell proliferation. Blocking the endogenous TNF-alpha activity with neutralizing anti-TNF-alpha antibodies abolished all these PMA-induced events with the exception of MMP-2 gene expression. Since fibronectin (FN)-mediated cell adhesion and spreading are prerequisite for both macrophage differentiation and MMP-9 gene expression in HL-60 cells, we hypothesized that TNF-alpha might be involved in modulating the expression of either the FN or its integrin receptor genes. Whereas PMA substantially enhanced the steady state mRNA and protein levels of both FN and alpha5 beta1 integrins, TNF-alpha alone had little effect on the expression of these genes. However, anti-TNF-alpha antibodies blocked PMA-induced augmentation of both alpha5 and beta1 integrin gene expression without affecting the expression of the FN gene. Our results suggest that TNF-alpha may regulate macrophage differentiation and critical matrix-degrading activities of myeloid progenitor cells in an autocrine manner by augmenting surface levels of the alpha5 beta1 integrin, thus promoting interactions with the extracellular matrix, a key event for maturation and migration of these cells during inflammation.


Comparison of heterologous prime-boost immunization strategies with DNA and recombinant vaccinia virus co-expressing GP3 and GP5 of European type porcine reproductive and respiratory syndrome virus in pigs.
Vaccination is principally used to control and treat porcine reproductive and respiratory syndrome virus (PRRSV) infection. This study investigated immunogenicity and protective efficacy of heterologous prime-boost regimens in pigs, including recombinant DNA and vaccinia virus vectors coexpressing PRRSV European genotype (EU) isolate GP3 and GP5: group A, pVAX1-EU-GP3-GP5 prime and rddVTT-EU-GP3-GP5 boost; group B, rddVTT-EU-GP3-GP5 prime and pVAX1-EU-GP3-GP5 boost; group C, empty vector pVAX1; group D, E3L gene-deleted vaccinia virus E3L- VTT. Vaccine efficacy was tested in an EU-type PRRSV (Lelystad virus strain) challenge pig model based on evaluating PRRSV-specific antibody responses, neutralizing antibodies, cytokines, T lymphocyte proliferation, CD4+ and CD8+ T lymphocytes, clinical symptoms, viremia and tissue virus loads. Plasmid DNA was delivered as chitosan-DNA nanoparticles, and Quil A (Quillaja) was used to increase vaccine efficiency. All piglets were boosted 21 days post the initial inoculation (dpi) and then challenged 14 days later. At 14, 21, 28 and 35 dpi, groups A and B developed significantly higher PRRSV-specific antibody responses compared with control groups C and D. Two weeks after the boost, significant differences in neutralizing antibody and IFN-γ levels were observed between groups A, C, D and B. At 49 dpi, groups A and B had markedly increased peripheral blood CD3+CD4+ T cell levels. Following virus challenge, group A showed viremia, but organ virus loads were lower than those in other groups. Thus, a heterologous prime-boost vaccine regimen (rddVTT-EU-GP3-GP5 prime, pVAX1-EU-GP3-GP5 boost) can improve humoral- and cell-mediated immune responses to provide resistance to EU-type PRRSV infection in vivo.


Development of Health Pathways to Standardize Cancer Care Pathways Informed by Patient-Reported Outcomes and Clinical Practice Guidelines.
High-quality symptom management and supportive care are essential components of comprehensive cancer care. We aimed to describe the development of an evidence-based automated decisional algorithm for patients with cancer that had specific, actionable, clinical, evidence-based recommendations to improve patient care, communication, and management. We reviewed existing literature and clinical practice guidelines to identify priority domains of patient care and potential clinical recommendations. Two multidisciplinary clinical advisory groups used a two-stage consensus decision-making approach to determine domains of care and patient-reported outcome (PRO) measures and subsequently developed automated algorithms with clear clinical recommendations amendable to intervention in clinical settings. Algorithms were developed to inform management of patient symptoms, distress, and unmet needs. Three PRO measures were chosen: Distress Thermometer and problem checklist, Edmonton Symptom Assessment Scale, and the Supportive Care Needs Survey-Screening Tool 9. PRO items were mapped to five domains of patient well-being: physical, emotional, practical, social and family, and maintenance of well-being. A total of 15 actionable clinical recommendations tailored to specific issues of concern were established. Using automated algorithms and clinical recommendations provides a platform for streamlining and systematizing the use of PROs to inform risk-stratified guideline-informed care. The series of algorithms, which set out systematized care pathways for the clinical care of patients with cancer, can be used to potentially inform patient-centered care.


Rapid morphological characterization of isolated mitochondria using Brownian motion.
Mitochondrial morphology has been associated with numerous pathologies including cancer, diabetes, obesity and heart disease. However, the connection is poorly understood-in part due to the difficulty of characterizing the morphology. This impedes the use of morphology as a tool for disease detection/monitoring. Here, we use the Brownian motion of isolated mitochondria to characterize their size and shape in a high throughput fashion. By using treadmill exercise training, mitochondria from heart and gastrocnemius of Balb/c mice were modulated in size and used to investigate the protocol. Consistent with previous reports, the heart mitochondria of untrained mice increased 5% in diameter immediately after a single bout of moderate exercise (1.091 ± 0.004 μm) as compared to completely sedentary controls (1.040 ± 0.022 μm). In addition, no change was observed in the size of gastrocnemius mitochondria (1.025 ± 0.018 μm), which was also in agreement with previous studies. The method was also successfully applied to smaller Saccharomyces cerevisiae mitochondria.


Patient-reported vision-related quality of life differences between superior and inferior hemifield visual field defects in primary open-angle glaucoma.
Previous studies have found that glaucoma is associated with impaired patient-reported vision-related quality of life (pVRQOL) but few, to our knowledge, have assessed how the visual field (VF) defect location impacts the pVRQOL. To investigate the associations of VF defects in the superior vs inferior hemifields with pVRQOL outcomes in patients with primary open-angle glaucoma. Prospective cross-sectional study at a tertiary referral center from March 1, 2012, to January 1, 2013, including patients with primary open-angle glaucoma who had a best-corrected visual acuity in the better eye equal to or better than 20/60 and reliable VF tests. The pVRQOL was assessed by a validated Taiwanese version of the 25-item National Eye Institute Visual Function Questionnaire. Reliable VF tests obtained within 3 months of enrollment were transformed to binocular integrated VF (IVF). The IVF was further stratified by VF location (superior vs inferior hemifield). The association between each domain of the 25-item National Eye Institute Visual Function Questionnaire and superior or inferior hemifield IVF was determined using multivariable linear regression analysis. The analysis included 186 patients with primary open-angle glaucoma with a mean age of 59.1 years (range, 19-86 years) and IVF mean deviation (MD) of -4.84 dB (range, -27.56 to 2.17 dB). In the multivariable linear regression analysis, the MD of the full-field IVF showed positive associations with near activities (β = 0.05; R2 = 0.20; P < .001), vision-specific role difficulties (β = 0.04; R2 = 0.19; P = .01), vision-specific dependency (β = 0.04; R2 = 0.20; P < .001), driving (β = 0.05; R2 = 0.24; P < .001), peripheral vision (β = 0.03; R2 = 0.18; P = .02), and composite scores (β = 0.04; R2 = 0.27; P = .005). Subsequent analysis showed that the MD of the superior hemifield IVF was associated only with near activities (β = 0.04; R2 = 0.21; P < .001) while the MD of the inferior hemifield IVF was associated with general vision (β = 0.04; R2 = 0.12; P = .01), vision-specific role difficulties (β = 0.04; R2 = 0.20; P = .01), and peripheral vision (β = 0.03; R2 = 0.17; P = .03). Superior hemifield IVF was strongly associated with difficulty with near activities. Inferior hemifield IVF impacted vision-specific role difficulties and general and peripheral vision. The impact of a VF defect on a patient's pVRQOL may depend not only on its severity, but also on its hemifield location.


A Pseudoscorpion's Promising Pinch: The venom of Chelifer cancroides contains a rich source of novel compounds.
With pedipalps modified for venom injection, some pseudoscorpions possess a unique venom delivery system, which evolved independently from those of other arachnids like scorpions and spiders. Up to now, only a few studies have been focused on pseudoscorpion venom, which either identified a small fraction of venom compounds, or were based on solely transcriptomic approaches. Only one study addressed the bioactivity of pseudoscorpion venom. Here, we expand existing knowledge about pseudoscorpion venom by providing a comprehensive proteomic and transcriptomic analysis of the venom of Chelifer cancroides. We identified the first putative genuine toxins in the venom of C. cancroides and we showed that a large fraction of the venom comprises novel compounds. In addition, we tested the activity of the venom at specific ion channels for the first time. These tests demonstrate that the venom of C. cancroides causes inhibition of a voltage-gated insect potassium channel (Shaker IR) and modulates the inactivation process of voltage-gated sodium channels from Varroa destructor. For one of the smallest venomous animals ever studied, today's toolkits enabled a comprehensive venom analysis. This is demonstrated by allocating our identified venom compounds to more than half of the prominent ion signals in MALDI-TOF mass spectra of venom samples. The present study is a starting point for understanding the complex composition and activity of pseudoscorpion venom and provides a potential rich source of bioactive compounds useable for basic research and industrial application.


Emergence of single-molecular chirality from achiral reactants.
The synthesis of enantiopure molecules from achiral precursors without the need for pre-existing chirality is a major challenge associated with the origin of life. We here show that an enantiopure product can be obtained from achiral starting materials in a single organic reaction. An essential characteristic of this reaction is that the chiral product precipitates from the solution, introducing a crystal-solution interface which functions as an asymmetric autocatalytic system that provides sufficient chiral amplification to reach an enantiopure end state. This approach not only provides more insight into the origin of life but also offers a pathway to acquire enantiopure compounds for industrial applications.


From empire to democracy: effects of social progress on Turkish writers.
What is the effect of a society's culture on the creative writers living there? Few cultures have had such a event as Ataturk's 1920 revolution that changed Turkey from a monarchy ruled by sultans into a republic. How would such a dramatic shift in a country's history be reflected in the accomplishments and characteristics of its writers? In this study, the authors investigated 948 eminent Turkish writers. Variables of gender, era, type of writing, education level, profession, and winning an award were all analyzed. The type of writing (fiction, poetry, plays, or nonfiction) and the era in which the writing was produced were both predictive of whether an author won a literary award. Before 1920, fiction writers received more awards than poets; after 1920, poets received more awards. In addition, professional writers were more likely to win awards than were professional politicians. Reasons for these findings are discussed with an emphasis on cultural and historical influences.


Heterotopic gastric mucosa of the rectum with a review of the literature.
Heterotopic gastric mucosa has been identified in all levels of the gastrointestinal tract but is a rare finding in the rectum. We describe a patient with heterotopia found during a routine evaluation of persistent symptoms, possibly due to an irritable bowel syndrome, and provide a complete review of the literature through 1992. Of the 29 reported cases, the most common presentation was painless rectal bleeding, with the heterotopia usually identified on pathologic examination of a rectal polyp. The heterotopic tissue was frequently located in the posterolateral region of the rectum between 5 and 8 cm from the anal verge. The definitive treatment was excision, either by endoscopy or surgery, although H2 may also provide symptom relief. Our patient was treated with H2 with resolution of symptoms.


[Role of clinical data in NST: from the viewpoint of the clinical laboratory].
Serum albumin (Alb) sampling is used for patients who object to screening by NST. As Alb is decreased by malnutrition, it is available for nutritional assessment, reflecting the long-term nutritive condition. Recently, Alb was correlated with nutritional assessment proteins with RTP, like RBP, TTR and Tf. We recently set up a screening line of Alb < or =3.0g/dl; however, we will gradually elevate it to < or =3.5g/dl. The role of the medical technologist is to estimate the number of patients with < or =3.5g/dl, and investigate correlations between the Alb concentration and other markers, including ChE, TTR, TP and CRP.


Decoding the Signals: An Analysis of Preference Signaling in the 2023 Orthopaedic Surgery Residency Match.
While previously used in other specialties, the preference signaling program (PSP) was implemented in the 2022 to 2023 orthopaedic surgery residency application process for the first time. The PSP allowed for 30 signaling tokens to be sent by applicants to programs of their choice to indicate particular interest in a program. It remains unknown how the PSP affects applicants and programs in the orthopaedic surgery residency match. Thus, this study's purpose was to assess the utility of preference signaling within the orthopaedic surgery residency application process in its inaugural year of use. An anonymous electronic survey was emailed to all orthopaedic surgery residency applicants who applied to the authors' institution during the 2022 to 2023 application cycle. The survey was sent after match lists were submitted and closed before the release of match results. The survey collected information regarding applicant demographics, preference signaling habits, and attitude toward preference signaling. The survey was completed by 101 applicants. Applicants applied to a mean of 90 programs (range: 10-197) and received an average of 12 interview invitations (range: 0-39). Applicants almost uniformly used all 30 signals, with nearly two-thirds signaling their home programs (65%, 49/76), and nearly all applicants sending signals to programs at which they performed away rotations (95.7%, 88/92). Applicants received a mean of 9 invitations from programs they signaled, compared with 2 invitations from programs they did not signal. Applicants were significantly more likely to receive an interview invite at a program they signaled than one they did not (p < 0.01). Overall, 57% of applicants (57/101) found the PSP to be helpful, whereas 28% (28/101) found it to be unhelpful, and 16% (16/101) had a neutral opinion. This study reports that the PSP in the 2022 to 2023 orthopaedic surgery match was an effective method of expressing interest in a program because applicants were significantly more likely to receive interview invites to signaled programs. More than half of respondents felt PSP to be helpful; however, the effect on application numbers is still unclear. III.


Inhibition of TLR4 Signalling-Induced Inflammation Attenuates Secondary Injury after Diffuse Axonal Injury in Rats.
Increasing evidence suggests that secondary injury after diffuse axonal injury (DAI) damages more axons than the initial insult, but the underlying mechanisms of this phenomenon are not fully understood. Recent studies show that toll-like receptor 4 (TLR4) plays a critical role in promoting adaptive immune responses and have been shown to be associated with brain damage. The purpose of this study was to investigate the role of the TLR4 signalling pathway in secondary axonal injury in the cortices of DAI rats. TLR4 was mainly localized in microglial cells and neurons, and the levels of TLR4 downstream signalling molecules, including TLR4, myeloid differentiation primary response gene 88, toll/IR-1-(TIR-) domain-containing adaptor protein inducing interferon-beta, interferon regulatory factor 3, interferon β, nuclear factor κB (NF-κB) p65, and phospho-NF-κB p65, significantly increased and peaked at 1 d after DAI. Inhibition of TLR4 by TAK-242 attenuated apoptosis, neuronal and axonal injury, and glial responses. The neuroprotective effects of TLR4 inhibition were associated with decreases in the levels of TLR4 downstream signalling molecules and inflammatory factors, including interleukin-1β, interleukin-6, and tumour necrosis factor-α. These results suggest that the TLR4 signalling pathway plays an important role in secondary injury and may be an important therapeutic target following DAI.


Infrared matrix-assisted laser desorption electrospray ionization mass spectrometry imaging analysis of biospecimens.
Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry imaging (MSI) is a technique well suited for analysis of biological specimens. This tutorial review focuses on recent advancements and applications of IR-MALDESI MSI to better understand key biological questions. Through optimization of user-defined source parameters, comprehensive and quantitative MSI data can be obtained for a variety of analytes. The effect of an ice matrix layer is well defined in the context of desorption dynamics and resulting ion abundance. Optimized parameters and careful control of conditions affords quantitative MSI data which provides valuable information for targeted, label-free drug distribution studies and untargeted metabolomic datasets. Challenges and limitations of MSI using IR-MALDESI are addressed in the context of the bioimaging field.


Rare events and discontinuous percolation transitions.
Percolation theory characterizing the robustness of a network has applications ranging from biology, to epidemic spreading, and complex infrastructures. Percolation theory, however, only concerns the average response of a network to random damage of its nodes while in real finite networks, fluctuations around this average behavior are observable. Consequently, for finite networks, there is an urgent need to evaluate the risk of collapse in response to rare configurations of the initial damage. Here, we build a large deviation theory of percolation characterizing the response of a sparse network to rare events. This general theory includes the second-order phase transition observed typically for random configurations of the initial damage, but reveals also discontinuous transitions corresponding to rare configurations of the initial damage for which the size of the giant component is suppressed.


Photodissociation dynamics of N3.
The photodissociation dynamics of N3 + excited from its (linear) 3Σg -/(bent) 3A″ ground to the first excited singlet and triplet states is investigated. Three-dimensional potential energy surfaces for the 1A', 1A″, and 3A' electronic states, correlating with the 1Δg and 3Πu states in linear geometry, for N3 + are constructed using high-level electronic structure calculations and represented as reproducing kernels. The reference ab initio energies are calculated at the MRCI+Q/aug-cc-pVTZ level of theory. For following the photodissociation dynamics in the excited states, rotational and vibrational distributions P(v') and P(j') for the N2 product are determined from vertically excited ground state distributions. Due to the different shapes of the ground state 3A″ potential energy surface and the excited states, appreciable angular momentum j' ∼ 60 is generated in diatomic fragments. The lifetimes in the excited states extend to at least 50 ps. Notably, results from sampling initial conditions from a thermal ensemble and from the Wigner distribution of the ground state wavefunction are comparable.


QTL Mapping of Seed Quality Traits Including Cooking Time, Flavor, and Texture in a Yellow Dry Bean (Phaseolus vulgaris L.) Population.
Manteca yellow dry beans (Phaseolus vulgaris L.) have many quality traits that appeal to consumers, including fast cooking times, creamy texture, and sweet, buttery flavor. They are native to Chile and consumed in regions of South America and Africa but are largely unfamiliar to United States consumers. While cooking time, flavor, and texture have not been prioritized in United States dry bean breeding programs, genetic variability exists such that these traits could be addressed through breeding. In this study, a recombinant inbred line (RIL) population was developed from a cross between Ervilha (Manteca) and PI527538 (Njano), yellow dry beans with contrasting cooking time and sensory attributes. The population and parents were grown for 2 years in Michigan and evaluated for cooking time and sensory attribute intensities, including total flavor, beany, vegetative, earthy, starchy, sweet, bitter, seed-coat perception, and cotyledon texture. Cooking time ranged 19-34 min and exhibited high broad-sense heritability (0.68). Sensory attribute intensities also exhibited variation among RILs, although broad-sense heritability was low, with beany and total flavor exhibiting the highest (0.33 and 0.27). A linkage map of 870 single nucleotide polymorphisms markers across 11 chromosomes was developed for quantitative trait loci (QTL) mapping, which revealed QTL for water uptake (3), cooking time (6), sensory attribute intensities (28), color (13), seed-coat postharvest non-darkening (1), seed weight (5), and seed yield (2) identified from data across 2 years. Co-localization was identified for starchy, sweet, and seed-coat perception on Pv01; for total flavor, beany, earthy, starchy, sweet, bitter, seed-coat perception, cotyledon texture, and color on Pv03; water uptake and color on Pv04; total flavor, vegetative, sweet, and cotyledon texture on Pv07; cooking time, starchy, sweet, and color on Pv08; and water uptake, cooking time, total flavor, beany, starchy, bitter, seed-coat perception, cotyledon texture, color, and seed-coat postharvest non-darkening on Pv10. The QTL identified in this work, in particular CT8.2 and CT10.2, can be used to develop molecular markers to improve seed quality traits in future dry bean varieties. Considering yellow dry beans already excel in quality and convenience, they might be an ideal market class to signal a new focus on consumer-valued traits in the United States.


Deep learning for the prediction of the chemotherapy response of metastatic colorectal cancer: comparing and combining H&E staining histopathology and infrared spectral histopathology.
Colorectal cancer is a global public health problem with one of the highest death rates. It is the second most deadly type of cancer and the third most frequently diagnosed in the world. The present study focused on metastatic colorectal cancer (mCRC) patients who had been treated with chemotherapy-based regimen for which it remains uncertainty about the efficacy for all eligible patients. This is a major problem, as it is not yet possible to test different therapies in view of the consequences on the health of the patients and the risk of progression. Here, we propose a method to predict the efficacy of an anticancer treatment in an individualized way, using a deep learning model constructed on the retrospective analysis of the primary tumor of several patients. Histological sections from tumors were imaged by standard hematoxylin and eosin (HE) staining and infrared spectroscopy (IR). Images obtained were then processed by a convolutional neural network (CNN) to extract features and correlate them with the subsequent progression-free survival (PFS) of each patient. Separately, HE and IR imaging resulted in a PFS prediction with an error of 6.6 and 6.3 months respectively (28% and 26% of the average PFS). Combining both modalities allowed to decrease the error to 5.0 months (21%). The inflammatory state of the stroma seemed to be one of the main features detected by the CNN. Our pilot study suggests that multimodal imaging analyzed with deep learning methods allow to give an indication of the effectiveness of a treatment when choosing.


Morpho-syntactic processing of Arabic plurals after aphasia: dissecting lexical meaning from morpho-syntax within word boundaries.
Within the domain of inflectional morpho-syntax, differential processing of regular and irregular forms has been found in healthy speakers and in aphasia. One view assumes that irregular forms are retrieved as full entities, while regular forms are compiled on-line. An alternative view holds that a single mechanism oversees regular and irregular forms. Arabic offers an opportunity to study this phenomenon, as Arabic nouns contain a consonantal root, delivering lexical meaning, and a vocalic pattern, delivering syntactic information, such as gender and number. The aim of this study is to investigate morpho-syntactic processing of regular (sound) and irregular (broken) Arabic plurals in patients with morpho-syntactic impairment. Three participants with acquired agrammatic aphasia produced plural forms in a picture-naming task. We measured overall response accuracy, then analysed lexical errors and morpho-syntactic errors, separately. Error analysis revealed different patterns of morpho-syntactic errors depending on the type of pluralization (sound vs broken). Omissions formed the vast majority of errors in sound plurals, while substitution was the only error mechanism that occurred in broken plurals. The dissociation was statistically significant for retrieval of morpho-syntactic information (vocalic pattern) but not for lexical meaning (consonantal root), suggesting that the participants' selective impairment was an effect of the morpho-syntax of plurals. These results suggest that irregular plurals forms are stored, while regular forms are derived. The current findings support the findings from other languages and provide a new analysis technique for data from languages with non-concatenative morpho-syntax.


Can 'abnormally' fertilized zygotes give rise to viable embryos?
Conventional practice in in vitro fertilization or intracytoplasmic sperm injection is to select the best quality embryos based on their morphology and cleavage status from a cohort of fertilized oocytes in which two pronuclei were observed at the time they were checked for fertilization. However, in a small proportion of cycles, the selection is limited to embryos that appeared to be either unfertilized (displaying zero pronuclei) or abnormally fertilized (displaying one or three pronuclei) at the time they were checked for fertilization. There is a lack of consensus on whether such embryos should be transferred to the uterus. Cytogenetic analysis of embryos from oocytes with one pronucleus has shown a proportion is diploid. Transfer of such embryos has resulted in healthy births. Limited cytogenetic analysis of oocytes that divide despite the absence of pronuclei at fertilization check indicates that a proportion also have a normal cytogenetic constitution. Cytogenetic analysis of embryos from oocytes with three pronuclei has shown high rates of triploidy and chaotic cell divisions. Subsequent foetuses have extremely unfavourable outcomes. Here, we review the published literature on the cytogenetic analysis of 'unfertilized' and 'abnormally fertilized' embryos and discuss possible pathways which lead to their formation. The limited evidence indicates that oocytes with one pronucleus and oocytes that show normal onward division despite the absence of pronuclei may be considered for replacement in certain circumstances.


Fragile X syndrome.
The fragile X syndrome is characterized by mental handicap, facial dysmorphism and expression of a fragile site at Xq27.3. An expansion of a CGG repeat in the 5' end of the fragile X mental retardation 1 (FMR1) gene results in the absence of the encoded fragile X mental retardation protein, known to play an important role in RNA processing and probably the developmental maturation of brain neurons.


The yeast ubiquitin genes: a family of natural gene fusions.
Ubiquitin is a 76-residue protein highly conserved among eukaryotes. Conjugation of ubiquitin to intracellular proteins mediates their selective degradation in vivo. We describe a family of four ubiquitin-coding loci in the yeast Saccharomyces cerevisiae. UB11, UB12 and UB13 encode hybrid proteins in which ubiquitin is fused to unrelated ('tail') amino acid sequences. The ubiquitin coding elements of UB11 and UB12 are interrupted at identical positions by non-homologous introns. UB11 and UB12 encode identical 52-residue tails, whereas UB13 encodes a different 76-residue tail. The tail amino acid sequences are highly conserved between yeast and mammals. Each tail contains a putative metal-binding, nucleic acid-binding domain of the form Cys-X2-4-Cys-X2-15-Cys-X2-4-Cys, suggesting that these proteins may function by binding to DNA. The fourth gene, UB14, encodes a polyubiquitin precursor protein containing five ubiquitin repeats in a head-to-tail, spacerless arrangement. All four ubiquitin genes are expressed in exponentially growing cells, while in stationary-phase cells the expression of UB11 and UB12 is repressed. The UB14 gene, which is strongly inducible by starvation, high temperatures and other stresses, contains in its upstream region strong homologies to the consensus 'heat shock box' nucleotide sequence. Elsewhere we show that the essential function of the UB14 gene is to provide ubiquitin to cells under stress.


High Refractive Index Polymer Thin Films by Charge-Transfer Complexation.
High refractive index polymers are essential in next-generation flexible optical and optoelectronic devices. This paper describes a simple synthetic method to prepare polymeric optical coatings possessing high refractive indexes. Poly(4-vinylpyridine) (P4VP) thin films prepared using initiated chemical vapor deposition are exposed to highly polarizable halogen molecules to form stable charge-transfer complexes: P4VP-IX (X = I, Br, and Cl). Fourier transform infrared spectroscopy was used to confirm the formation of charge-transfer complexes. Characterized by spectroscopic ellipsometry, the maximum refractive index of 2.08 at 587.6 nm is obtained for P4VP-I2. For P4VP-IBr and P4VP-ICl, the maximum refractive indexes are 1.849 and 1.774, respectively. By controlling the concentration of charge-transfer complexes, either through the halogen incorporation step or polymer composition through copolymerization with ethylene glycol dimethacrylate, the refractive indexes of the polymer thin films can be precisely controlled. The feasibility of P4VP-IX materials as optical coatings is also explored. The refractive index and thickness uniformity of a P4VP-I2 film over a 10 mm diameter circular area were characterized, showing standard deviations of 0.0769 and 1.91%, respectively.


Cell culture systems for hepatitis C virus.
Due to the obligatory intracellular lifestyle of viruses, cell culture systems for efficient viral propagation are crucial to obtain a detailed understanding of the virus-host cell interaction. For hepatitis C virus (HCV) the development of permissive and authentic culture models continues to be a challenging task. The first efforts to culture HCV had limited success and range back to before the virus was molecularly cloned in 1989. Since then several major breakthroughs have gradually overcome limitations in culturing the virus and sequentially permitted analysis of viral RNA replication, cell entry, and ultimately the complete replication cycle in cultured cells in 2005. Until today, basic and applied HCV research greatly benefit from these tremendous efforts which spurred multiple complementary cell-based model systems for distinct steps of the HCV replication cycle. When used in combination they now permit deep insights into the fascinating biology of HCV and its interplay with the host cell. In fact, drug development has been much facilitated and our understanding of the molecular determinants of HCV replication has grown in parallel to these advances. Building on this groundwork and further refining our cellular models to better mimic the architecture, polarization and differentiation of natural hepatocytes should reveal novel unique aspects of HCV replication. Ultimately, models to culture primary HCV isolates across all genotypes may teach us important new lessons about viral functional adaptations that have evolved in exchange with its human host and that may explain the variable natural course of hepatitis C.


Beta-glucanases of the yeast Pichia polymorpha.
Fractionation of proteins secreted into the culture medium by intact cells and protoplasts of Pichia polymorpha showing enzyme activity against laminarin, pustulan or p-nitrophenyl-beta-D-glucopyranoside has been performed, and the results compared with those obtained with cell-free extracts and lysed protoplasts. Fractionation with DEAE Sephadex A50 has proved to be the best method, yielding at least three fractions which hydrolyse laminarin. One of these fractions was active on both laminarin and pustulan. Filtration on Sephadex G-100 column only yielded one active preparation. Evidence supporting the conclusion that there are three different beta-glucanases located in the periplasmic space is presented.


Seeded QED cascades in counterpropagating laser pulses.
The growth rates of seeded QED cascades in counterpropagating lasers are calculated with first-principles two- and three-dimensional QED-PIC (particle-in-cell) simulations. The dependence of the growth rate on the laser polarization and intensity is compared with analytical models that support the findings of the simulations. The models provide insight regarding the qualitative trend of the cascade growth when the intensity of the laser field is varied. A discussion about the cascade's threshold is included, based on the analytical and numerical results. These results show that relativistic pair plasmas and efficient conversion from laser photons to γ rays can be observed with the typical intensities planned to operate on future ultraintense laser facilities such as ELI or Vulcan.


Beneficial Effects of Exercise on Subendothelial Matrix Stiffness are Short-Lived.
Aerobic exercise helps to maintain cardiovascular health in part by mitigating age-induced arterial stiffening. However, the long-term effects of exercise regimens on aortic stiffness remain unknown, especially in the intimal extracellular matrix layer known as the subendothelial matrix. To examine how the stiffness of the subendothelial matrix changes following exercise cessation, mice were exposed to an 8 week swimming regimen followed by an 8 week sedentary rest period. Whole vessel and subendothelial matrix stiffness were measured after both the exercise and rest periods. After swimming, whole vessel and subendothelial matrix stiffness decreased, and after 8 weeks of rest, these values returned to baseline. Within the same time frame, the collagen content in the intima layer and the presence of advanced glycation end products (AGEs) in the whole vessel were also affected by the exercise and the rest periods. Overall, our data indicate that consistent exercise is necessary for maintaining compliance in the subendothelial matrix.