Downregulation of low-density lipoprotein receptor mRNA in lymphatic endothelial cells impairs lymphatic function through changes in intracellular lipids.
Rationale: Impairment in lymphatic transport is associated with the onset and progression of atherosclerosis in animal models. The downregulation of low-density-lipoprotein receptor (LDLR) expression, rather than increased circulating cholesterol level per se, is involved in early atherosclerosis-related lymphatic dysfunction. Enhancing lymphatic function in Ldlr-/- mice with a mutant form of VEGF-C (VEGF-C 152s), a selective VEGFR-3 agonist, successfully delayed atherosclerotic plaque onset when mice were subsequently fed a high-fat diet. However, the specific mechanisms by which LDLR protects against lymphatic function impairment is unknown. Methods and results: We have thus injected wild-type and Pcsk9-/- mice with an adeno-associated virus type 1 expressing a shRNA for silencing Ldlr in vivo. We herein report that lymphatic contractility is reduced upon Ldlr dowregulation in wild-type mice only. Our in vitro experiments reveal that a decrease in LDLR expression at the mRNA level reduces the chromosome duplication phase and the protein expression of VEGFR-3, a membrane-bound key lymphatic marker. Furthermore, it also significantly reduced the levels of 18 lipid subclasses, including key constituents of lipid rafts as well as the transcription of several genes involved in cholesterol biosynthesis and cellular and metabolic processes. Exogenous PCSK9 only reduces lymphatic endothelial-LDLR at the protein level and does not affect lymphatic endothelial cell integrity. This puts forward that PCSK9 may act upon lymphatic muscle cells to mediate its effect on lymphatic contraction capacity in vivo. Conclusion: Our results suggest that treatments that specifically palliate the down regulation of LDLR mRNA in lymphatic endothelial cells preserve the integrity of the lymphatic endothelium and sustain lymphatic function, a prerequisite player in atherosclerosis.


Changes in peripapillary retinal nerve fiber layer distribution by axial length.
To investigate the effect of axial length on the peripapillary retinal nerve fiber layer (RNFL) distribution and the risk of misdiagnosing normal subjects when using the instrument's built-in normative database. Healthy young volunteers underwent ophthalmologic examinations including RNFL thickness analysis with spectral-domain optical coherence tomography (OCT), autorefraction, and axial length measurement. The average RNFL thickness values reported along the calculation circle for whole circle, quadrants, and clock hours were recorded and evaluated at the 5% probability level for significant disparities in comparison with the built-in RNFL database. Angular locations with maximal RNFL thickness in the superotemporal and inferotemporal RNFL humps were determined. The relationships of axial length with these variables were analyzed using both linear regression and logistic regression models. A total of 485 eyes of 485 subjects were analyzed. The average 360° RNFL thickness became thinner as the axial length increased (r = -0.244, p < 0.001). As the axial length increased, the average RNFL thicknesses in the 1, 2, 4, 5, 6, and 12-o'clock hour sectors decreased whereas those of the 7, 8, 9, 10, and 11-o'clock hour sectors increased. In addition, as the axial length increased, two major RNFL humps were located more temporally and it became significantly more likely for the 1, 2, 5, 6, and 12-o'clock hour sectors to fall below the p < 0.05 significance limits for normal thickness. Peripapillary RNFL thickness changes differently in different peripapillary locations as axial length increases. Because long eyes may lead to misleading findings of OCT parameters depending on the peripapillary location, their relationship to the axial length should be considered in the built-in RNFL thickness normative database of OCT.


Integration of Facial Expression and Gaze Direction in Individuals with a High Level of Autistic Traits.
We investigated whether individuals with high levels of autistic traits integrate relevant communicative signals, such as facial expression, when decoding eye-gaze direction. Students with high vs. low scores on the Autism Spectrum Quotient (AQ) performed a task in which they responded to the eye directions of faces, presented on the left or the right side of a screen, portraying different emotional expressions. In both groups, the identification of gaze direction was faster when the eyes were directed towards the center of the scene. However, in the low AQ group, this effect was larger for happy faces than for neutral faces or faces showing other emotional expressions, whereas participants from high AQ group were not affected by emotional expressions. These results suggest that individuals with more autistic traits may not integrate multiple communicative signals based on their emotional value.


Studies of Coquillettidia (coquillettidia) crassipes (Vanderwulp, 1881) in relation to transmission of Cardiofilaria nilesi.
Field surveillance of Cq. crassipes was conducted in an open housing estate near Kuala Lumpur using IMR traps baited with chicken and gerbils. Chicken bait attracted more Cq. crassipes. There was significant difference between chicken and gerbil as bait. The mosquitoes preferred to feed at canopy level. The parous rate was 20.41% and the infection rate for Cardiofilaria nilesi was 2.04%. None of the 120 chicken and 5 ducks, in a village close to the open housing estate, examined for microfilariae were positive. Ar. durhami and Ar. subalbatus did not support development of C. nilesi. However, Cq. crassipes is an extremely efficient vector of C. nilesi.


A resounding success or a disastrous failure: re-examining the interpretation of evidence on the Portuguese decriminalisation of illicit drugs.
In this Harm Reduction Digest two observers and scholars of the 2001 Portuguese drug policy reform consider divergent accounts of the reform which viewed it as a 'resounding success' or a 'disastrous failure'. Acknowledging from their own experience the inherent difficulties in studying drug law reform, Caitlin Hughes and Alex Stevens take the central competing claims of the protagonists and consider them against the available data. They remind us of the way all sides of the drug policy debates call upon and alternatively use or misuse 'evidence' to feed into discussions of the worth, efficacy and desirability of different illicit drug policies. In doing so they provide pause for thought for those of us who operate as drug policy researchers and drug policy advocates.


Microporous modified atmosphere packaging to extend shelf life of fresh foods: A review.
In recent years, microporous modified atmosphere packaging has been widely concerned because of its adjustable air permeability and low processing cost. With the development and increasing demand of fresh food industry, the limited permeability of film in modified atmosphere packaging can't meet the fresh-keeping requirements of fresh foods, especially vegetables and fruits. Microporous film can flexibly adjust the gas permeability according to the physiological metabolic characteristics of fresh foods, which has gradually become a fresh-keeping technology in the domain of vegetables and fruits. This paper reviewed the research progress of microporous modified atmosphere packaging and its extension on shelf life of fresh foods. The latest applied researches were described in a comprehensive manner, particularly fruits and vegetables. Besides, this article also covered theoretical support and analysis, including the perforation mode, air permeability mechanism and mathematical model of microporous film, the characteristics of fresh foods, pore parameters and traits of film materials. This paper payed attention to the application of environmentally friendly degradable film materials (biological film materials, nano materials) in fruits and vegetables preservation. Research has shown that the degradable material can enlarge the fresh-keeping effect of microporous modified atmosphere packaging, which is worthy of further research and development. Finally, the development trends and directions in the future were discussed.


Laser annealing to improve PbSe thin film photosensitivity and specific detectivity.
PbSe thin films were deposited on SiO2/Si wafers using chemical bath deposition for mid-wave infrared (MWIR) detection. To enhance the photosensitivity of PbSe thin films, oxidation, followed by iodization, was performed to create a PbI2/PbSe two-layer system for efficient MWIR detection in the spectral range from 3 µm to 5 µm. A near-infrared (IR) laser annealing was performed after sensitization with 1070 nm wavelength at an energy density of 1J/cm2 to selectively heat the PbSe thin films. After IR laser annealing, the change in resistance between dark condition and MWIR illumination improved significantly from 19.8% to 22.6%. In addition, the dark resistance increased by 32.5% after IR laser annealing. IR photoluminescence spectra after IR laser annealing shows an increase in the sub-peak intensities from iodine incorporation. The results indicate that more iodine is incorporated into Se sites at the outer regions of PbSe grains. Therefore, more donors (electrons) from iodine diffuse into PbSe and recombine with holes so that PbSe thin film after IR laser annealing shows much higher dark resistance. Test devices with NiCr electrodes at the bottom of PbSe were fabricated with feature sizes of 40 µm to investigate the effect of IR laser annealing on electrical properties and specific detectivity (D∗). I-V characteristics show dark resistance increased after IR laser annealing. The specific detectivity increases significantly after IR laser annealing at the applied bias of 10 V at 270 K from 0.55×1010cmHz1/2W-1 to 1.23×1010cmHz1/2W-1 due to dramatic noise reduction, which is originated from higher dark resistance.


Adenoid cystic carcinoma of the superonasal conjunctival fornix.
The case of a 32-year-old female with a mass in the superonasal conjunctival fornix is described. Histological examination revealed an adenoid cystic carcinoma, which we consider to be most likely derived from an accessory lacrimal gland. The clinical presentation, histological features, management and follow-up are described. A literature review of orbital adenoid cystic carcinoma arising outside the main lacrimal gland is presented.


Delivery of HFA and CFC salbutamol from spacer devices used in infancy.
The aim of the study was to compare the in vitro delivery of four salbutamol pressurized metered-dose inhalers (pMDIs) via the three spacer devices commonly used in European infants: Aerochamber-Infant, Babyhaler, and metallic NES-spacer. Emitted dose (ED) and fine particle dose (FPD, particles<5.8 microm) of each combination of spacer device and pMDI (chlorofluorocarbon-based Ventoline, Eolène, Spréor, and hydrofluoroalkane-based Airomir were measured respectively using unit dose sampling tubes (n=30 per combination) and an 8-stage cascade impactor (n=6 per group). The results were compared by analysis of variance and the Student-Newman-Keuls method. ED of Airomir was always greater than for Ventoline (P<0.05). FPD obtained with Ventoline was the lowest, with Eolène>Airomir=Spréor>Ventoline (P<0.05). Only Airomir produced a similar FPD with all three spacer devices. Chlorofluorocarbon-salbutamol pMDIs are not generics when used with spacer devices. The three spacer devices may be used interchangeably with Airomir.


Diagnosis and treatment of bacterial pneumonia in liver transplantation recipients: report of 33 cases.
Bacterial pneumonia in the recipients of liver transplantation (LTX) is a common postoperative complication influencing the prognosis greatly. In this article, the diagnosis and treatment of bacterial pneumonia in 33 LTX recipients are reported. From February 1999 to January 2003, a total of 103 patients underwent allogeneic LTX at our center; afterwards, a retrospective analysis was made on their postoperative clinical manifestations, including symptoms (expectoration, panting and fever), sign (rale), results of laboratory examinations (white blood cell count and sputum culture of tracheal secretions or pleural fluid culture), and chest X-ray films. The following data of the pneumonia and non-pneumonia groups were collected, and the rank sum test (SPSS 11.0, Wilcoxon's method) was used to analyze the duration of postoperative respirator utilization and the volume of pleural effusion through pleurocentesis or pleural drainage. In the 103 patients, 33 experienced 53 episodes of bacterial pneumonia during their hospital stay after transplantation, 14 of them (42.42%) had more than three manifestations of the seven mentioned above. The pathogens causing bacterial pneumonia included Pseudomonas aeruginosa (17.48%), Klebsiella pneumoniae (15.53%), Acinetobacter baumannii (10.68%), and Staphylococcus aureus (7.77%). Amilkacin, tienam, ciprofloxacin, vancomycin, etc. were the antibiotics of choice against those bacteria. Acute rejection occurred during the treatment of bacterial pneumonia in 16 patients, and 5 of them died. Wilcoxon's rank sum test of the data indicated that the pneumonia group had longer duration of postoperative ventilator treatment and larger volume of pleural effusion than the non-pneumonia group (P < 0.05). The clinical manifestations of pneumonia after LTX might be atypical, and special attention should be paid to the respiratory symptoms and signs within 2 months after LTX. Whenever the diagnosis of bacterial pneumonia is confirmed, consideration should be given to reasonable use of antibiotics and regulation of immunity in addition to other routine therapies.


The roles for cytokines in the generation and maintenance of regulatory T cells.
As an essential mechanism for self-tolerance, immune suppression has attracted much attention since the discovery of suppressor T cells, now called regulatory T cells (Tregs), in the 1990s. Different types of Tregs have been described based on distinct expression patterns of surface markers and cytokines. Cytokines are not only essential for function but also important for the generation of Tregs. Interleukin-2 (IL-2), transforming growth factor-beta, IL-10, and other immunoregulatory molecules have been shown to control the generation of Tregs. The presence of other types of cells, in particular antigen-presenting cells (APCs), is critical for the generation of Tregs. Cytokines can serve as either initiators or intermediates for the interactions between APCs and Tregs. This review discusses our current knowledge of how cytokines regulate the generation and maintenance of Tregs.


Reconceiving delusion.
Delusions are critical components in a number of mental disorders, schizophrenia foremost. What are they? The standard view is that they are a type of belief--a pathological belief. Unfortunately, the standard view does not consistently correspond to clinical practice, where the term 'delusion' often applies to non-beliefs. We review the case for saying that non-beliefs can count as delusions. We argue that delusions are complexes of higher and lower order attitudes. They constitute a distinctive type of failure of self-knowledge and self-management. We describe the relevant type. One of the conceptual implications of our view is that beliefs need not be central to delusions.


Molecular detection of Treponema species organisms in foremilk and udder cleft skin of dairy cows with digital dermatitis.
Identification of reservoirs and transmission routes of digital dermatitis (DD)-associated Treponema spp. is considered an effective means for controlling DD infection in dairy cows. The objective of this study is to identify and characterize the potential reservoir niches for DD-associated Treponema spp. from healthy udder cleft skin and foremilk in lactating dairy cows. A large dairy farm was visited weekly from March to July 2015. Clinical investigation revealed that a total of 25 lame cows had DD lesions located at the plantar aspect of the interdigital cleft. A total of 75 samples, three per cow, were collected including deep swabs from DD lesions (n = 25), non-aseptically collected foremilk samples (n = 25) and skin swabs from udder cleft (n = 25). Treponema spp. were identified using nested PCR assays and confirmed by DNA sequencing. Results revealed that Treponema phagedenis (T. phagedenis)-like was the most identified species in the foremilk 40% (10/25), in comparison with DD lesions and udder cleft skin samples with 32% (8/25) and 20% (5/25), respectively. On the other hand, Treponema pedis (T. pedis) was the most identified species in the udder cleft skin 80% (20/25), in comparison with DD lesions and foremilk samples with 68% (17/25) and 60% (15/25), respectively. None of the examined samples were identified by PCR as containing DNA from Treponema medium (T. medium) or Treponema vincentii (T. vincentii)-like. To the best of our knowledge, this is the first report for detection of T. phagedenis-like and T. pedis from healthy skin of udder cleft and foremilk samples. Detection of DD Treponema spp. from udder cleft skin and foremilk samples indicates that these sites could be potential reservoirs for spirochetes involved in DD. Udder cleft skin and foremilk may have a role in transmission routes of DD Treponema in dairy farms.


Results in the management of locally unresectable pancreatic carcinoma.
Between 1973 and 1983, 43 patients with histologically proven unresectable pancreatic carcinoma were irradiated in the UCLA Department of Radiation Oncology. Ten patients received irradiation alone and 33 were nonrandomly assigned to receive chemotherapy in addition to irradiation. Of those patients receiving chemotherapy, 30 were given 5-fluorouracil and three were given a combination of agents. Forty-one of the 43 patients have died with a median survival of 7 months. Actuarial survival at 1 and 2 years was 24% and 3%. Local control was achieved in three of 43 patients. Two patients are alive with no evidence of disease at 11 and 30 months. The median survivals with and without chemotherapy were 9.5 and 4 months, respectively (p = 0.06). Survival dependent on nodal status, surgical bypass, primary site, and dose are also reported. No significant differences were found. Acute complications were noted in 23 patients but were a reason for discontinuing therapy in none. Late complications were noted in nine patients. Six patients with an upper gastrointestinal hemorrhage or a small bowel obstruction all had local recurrence. There were two patients with posttreatment diabetes mellitus and one with pancreatitis. The limits of conventional therapy for unresectable pancreatic cancer have been reached. Creative sequencing of induction combination chemotherapy, newer radiation modalities, and maintenance chemotherapy are required if systemic and local progression of this lethal disease is to be eliminated.


[Historical observations on shoulder dystocia. 1) From the origin to Jacquemier].
The result of a literature survey on shoulder dystocia from year 1609 to 1860 are presented. Operative maneuvers for the resolution of dystocia were mainly examined, as well as the development of knowledge about the pathogenetic mechanism of this serious complication.


In situ monitoring of soil dissolution dynamics: a rapid and simple method for determining worst-case soils for cleaning validation.
A common approach utilized in validating the cleaning of multi-product equipment is to challenge the cleaning cycle with the hardest-to-clean, or "worst-case", soil. The worst-case soil is often determined through bench-scale experiments. These experiments can be tricky and time-consuming, especially when the dissolution dynamics of the soils are characterized by crossovers beyond the limit of visual detection. This paper describes a simple in situ method for monitoring soil dissolution dynamics and rapidly determining worst-case soils for cleaning validation. The method utilizes the real-time response of electrical conductivity to identify crossovers and to determine the optimal time for endpoint analysis. The results address the sensitivity and reproducibility of the method and the identification of critical experimental parameters. The dissolution dynamics of several parenteral products are compared.


Antifungal triazoles and polymorphonuclear leukocytes synergize to cause increased hyphal damage to Scedosporium prolificans and Scedosporium apiospermum.
Scedosporium prolificans and Scedosporium apiospermum (Pseudallescheria boydii) cause pulmonary and disseminated infections refractory to most currently used antifungal agents in immunocompromised patients. We therefore investigated the potential antifungal activities of the triazoles itraconazole (ITC), voriconazole (VRC), and posaconazole (PSC) in combination with human polymorphonuclear leukocytes (PMNs) against the hyphae of these fungal pathogens. A colorimetric assay with (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]2H-tetrazolium-5-carboxanilide) sodium salt was used for the measurement of hyphal damage as an indicator of antifungal activity. We found that the newer triazoles VRC and PSC displayed synergistic effects with PMNs against S. prolificans hyphae after 24 h (P < 0.05), whereas the effect of ITC in combination with PMNs was additive (P < 0.01). All three triazoles displayed additive antifungal activities in combination with PMNs against S. apiospermum hyphae (P < 0.05). The synergistic or additive effects that these triazoles exhibited, combined with the antifungal activities of human PMNs, may have important therapeutic implications for the management of infections due to S. prolificans and S. apiospermum.


Can we teach core clinical obstetrics and gynaecology skills using low fidelity simulation in an interprofessional setting?
Core clinical skills acquisition is an essential component of undergraduate medical and midwifery education. Although interprofessional education is an increasingly common format for learning efficient teamwork in clinical medicine, its value in undergraduate education is less clear. We present a collaborative effort from the medical and midwifery schools of Monash University, Melbourne, towards the development of an educational package centred around a core skills-based workshop using low fidelity simulation models in an interprofessional setting. Detailed feedback on the package was positive with respect to the relevance of the teaching content, whether the topic was well taught by task trainers and simulation models used, pitch of level of teaching and perception of confidence gained in performing the skill on a real patient after attending the workshop. Overall, interprofessional core skills training using low fidelity simulation models introduced at an undergraduate level in medicine and midwifery had a good acceptance.


DFT Studies on cis-1,4-Polymerization of Dienes Catalyzed by a Cationic Rare-Earth Metal Complex Bearing an Ancillary PNP Ligand.
Dnsity functional theory (DFT) calculations have been carried out for the highly selective cis-1,4-polymerization of butadiene catalyzed by a cationic rare-earth metal complex bearing an ancillary PNP ligand. It has been found that the chain initiation and propagation of butadiene polymerization occurs via the favorable cis-1,4-insertion route. The trans-1,4 and 1,2-insertion are unfavorable both kinetically and thermodynamically. The chain growth follows the π-allyl-insertion mechanism. The analyses of energy decomposition of transition states indicate that the likelihood of rival insertion pathways is predominantly controlled by the interaction energy of butadiene with a metal center and the deformation energy of butadiene moiety. The electronic factor of the central metal has a decisive influence on the cis- vs. trans-insertion and the regioselectivity (cis-1,4- vs. cis-1,2-insertion) is mainly determined by steric hindrance. Tetrahydrofuran (THF) coordination made monomer insertion less favorable compared with THF-free case and had more noticeable impact on the trans-monomer insertion compared with the cis case. During the chain propagation, cis-insertion of monomer facilitates THF de-coordination and the THF molecule could therefore dissociate from the central metal.


Distortion of aortic valve from mechanical traction imposed by the mitral valve prosthesis: The three-dimensional transesophageal echocardiographic perception.
Iatrogenic injury to the aortic valve is a rare but frequently reported complication during mitral valve surgeries. Intraoperative 2-dimensional transesophageal echocardiography (2D TEE) has a major impact in diagnosing these injuries, so that timely intervention is possible. However, 2D TEE has lot of limitations during the perioperative period, which can be overcome by the three dimensional echocardiography (3D-TEE). We report a case where 3D TEE has undoubtedly delineated the cause for distortion of aortic sinus after mitral valve replacement and helped in the successful outcome.


Privacy Act of 1974; systems of records--HCFA. Notice of proposed new routine uses for existing systems of records.
One of the top priorities of the Department of Health and Human Services is to assure high quality and effective health care while pursuing strategies to contain or moderate health care costs and Medicare program expenditures. One such program to limit Medicare programs expenditures was the Medicare Secondary payer (MSP) provisions (42 U.S.C. 1395y(b)). The purpose of this routine use is to enable HCFA and other "entities responsible for payment" to engage in exchanges of information concerning primary or secondary responsibility for a Medicare beneficiary's health care expenses. This notice is not the basis for a mandatory reporting requirement for "entities responsible for making payment" as required by 42 U.S.C. 1395y(b). Currently no statute mandates this type of exchange of information, therefore, these exchanges of information are strictly voluntary. The Health Care Financing Administration (HFCA) is amending systems notices for (1) Carrier Medicare Claims Records HHS/HCFA/BOP No. 09-70-0501; (2) Health Insurance Master Record HHS/HCFA/BOP No. 09-70-0502 and (3) Intermediary Medicare Claims Records HHS/HCFA/BPO No. 09-70-0503, to add a new routine use. The proposed routine use will enhance our capability for identifying Medicare Secondary payer situations where Medicare can assume a position of reduced liability due to the presence of other entities responsible for making primary payment in accordance with 42 U.S.C. 1395y(b).


Sheep pulmonary adenomatosis: characterization of two pathological forms associated with jaagsiekte retrovirus.
Pathological and immunohistochemical studies were performed on the lungs of 10 sheep with lesions of "classical" sheep pulmonary adenomatosis (SPA) and six sheep with "atypical" lung tumours. Lung tumour samples and other tissues from the same 16 animals were tested for the presence of jaagsiekte retrovirus (JSRV) by a polymerase chain reaction (PCR) that amplified a portion of the U3 long terminal repeat. The differences in the gross appearance of the classical and atypical forms paralleled the histopathological differences. The latter mainly concerned the stroma of the tumours which in the atypical cases was more heavily infiltrated by inflammatory cells and connective tissue fibres. JSRV major capsid protein was detected immunohistochemically in the epithelial transformed cells of both classical and atypical tumours, but the immune reactivity was slightly milder in atypical SPA. Proviral U3 sequences of JSRV were detected by specific PCR in all the tumour samples. Furthermore, the sequences of amplimers obtained from the two different pathological forms of the tumour were very similar. However, the dissemination of JSRV to other organs was greater in sheep with classical SPA than in those with atypical SPA. The pathological and virological features of these two forms of tumour are compared in an attempt to clarify whether classical and atypical SPA are two separate diseases or different expressions of a single disease spectrum.


Accelographic train-of-four at near-threshold currents.
The authors evaluated train-of-four (TOF) fade, as quantified by accelography, in response to neurostimulation at currents ranging from 10 to 60 mA. This was done to determine the range of currents over which measurements of fade remain consistent. In 31 patients (ASA Physical Status 1,2, and 3), anesthesia was induced with fentanyl, midazolam, and thiopental and was maintained with isoflurane and 66% nitrous oxide in oxygen. Surface stimulating electrodes were placed over the ulnar nerve, and an acceleration transducer was placed on the thumb. Succinylcholine was administered to facilitate tracheal intubation; after neuromuscular recovery, a bolus of vecuronium (0.01-0.05 mg.kg-1) and an infusion (0.25-1.5 micrograms.kg-1.min-1) were administered. After documentation of a stable TOF ratio, accelographic TOF responses were quantified in response to 200-microseconds stimulation at 10, 15, 20, 30, 40, 50, and 60 mA, in random order. A total of 95 data sets were collected at different depths of blockade. The TOF ratios maintained intercurrent consistency (P = not significant by nonparametric repeated measures analysis of variance), except at currents near the fourth-twitch (T4) threshold current. This inconsistency was eliminated by testing at greater than or equal to 10 mA above threshold. TOF ratios obtained at 10 mA above T4 threshold correlated highly with those at 60 mA (Spearman r value = 0.94). The authors conclude that the TOF ratio is consistent over a wide range of stimulating currents and that testing with submaximal currents can be performed reliably at greater than or equal to 10 mA above the T4 threshold.(ABSTRACT TRUNCATED AT 250 WORDS)


Efficacy and safety of recombinant follicle stimulating hormone (Puregon) in infertile women pituitary-suppressed with triptorelin undergoing in-vitro fertilization: a prospective, randomized, assessor-blind, multicentre trial.
The objective of this study was to compare the efficacy and safety of a recombinant follicle stimulating hormone (FSH) preparation (Org 32489, Puregon) with a urinary FSH preparation (Metrodin) in infertile women undergoing in-vitro fertilization (IVF and embryo transfer and who were pituitary-suppressed with triptorelin. In an assessor-blind, group-comparative, multicentre study, 60 women were randomized to Org 32489 and 39 to urinary FSH. An evaluation of the main parameter, the mean total number of oocytes recovered, indicated a similar efficacy for the two preparations: 9.7 with Org 32489 versus 8.9 with urinary FSH. In addition, there were no significant between-group differences with respect to other efficacy variables such as the total dose used, the duration of the treatment, the number of follicles > or = 17 mm in diameter and embryo quality. The ongoing pregnancy rates per attempt (30.2 versus 17.4%) and per transfer (34.0 versus 18.8%) were higher with Org 32489, but this difference was not statistically significant. No clinically relevant differences between Org 32489 and urinary FSH were seen with respect to safety variables. Serum antibodies were not detected in any of the subjects. It is concluded that Org 32489 compares favourably with urinary FSH in the treatment of infertile pituitary-suppressed women undergoing IVF and embryo transfer.


A Review of Strawberry Photobiology and Fruit Flavonoids in Controlled Environments.
Rapid technology development in controlled environment (CE) plant production has been applied to a large variety of plants. In recent years, strawberries have become a popular fruit for CE production because of their high economic and nutritional values. With the widespread use of light-emitting diode (LED) technology in the produce industry, growers can manipulate strawberry growth and development by providing specific light spectra. Manipulating light intensity and spectral composition can modify strawberry secondary metabolism and highly impact fruit quality and antioxidant properties. While the impact of visible light on secondary metabolite profiles for other greenhouse crops is well documented, more insight into the impact of different light spectra, from UV radiation to the visible light spectrum, on strawberry plants is required. This will allow growers to maximize yield and rapidly adapt to consumer preferences. In this review, a compilation of studies investigating the effect of light properties on strawberry fruit flavonoids is provided, and a comparative analysis of how light spectra influences strawberry's photobiology and secondary metabolism is presented. The effects of pre-harvest and post-harvest light treatments with UV radiation and visible light are considered. Future studies and implications for LED lighting configurations in strawberry fruit production for researchers and growers are discussed.


Tunable Plasmonic Cavity for Label-free Detection of Small Molecules.
Owing to its high sensitivity and high selectivity along with rapid response time, plasmonic detection has gained considerable interest in a wide variety of sensing applications. To improve the fieldwork applicability and reliability of plasmonic detection, the integration of plasmonic nanoparticles into optical devices is desirable. Herein, we propose an integrated label-free detection platform comprising a plasmonic cavity that allows sensitive molecular detection via either surface-enhanced Raman scattering (SERS) or plasmon resonance energy transfer (PRET). A small droplet of metal ion solution spontaneously produces a plasmonic cavity on the surface of uncured poly(dimethylsiloxane) (PDMS), and as PDMS is cured, the metal ions are reduced to form a plasmonic antennae array on the cavity surface. Unique spherical feature and the integrated metallic nanoparticles of the cavity provide excellent optical functions to focus the incident light in the cavity and to rescatter the light absorbed by the nanoparticles. The optical properties of the plasmonic cavity for SERS or PRET are optimized by controlling the composition, size, and density of the metal nanoparticles. By using the cavity, we accomplish both 1000-fold sensitive detection and real-time monitoring of reactive oxygen species secreted by live cells via PRET. In addition, we achieve sensitive detection of trace amounts of toxic environmental molecules such as 5-chloro-2-methyl-4-isothiazolin-3-one/2-methyl-4-isothiazol-3-one (CMIT/MIT) and bisphenol A, as well as several small biomolecules such as glucose, adenine, and tryptophan, via SERS.


Prevalence of scoliosis in beta-thalassemia.
The objective of this study was to determine the prevalence and possible pathogenesis of scoliosis in beta-thalassemia in our country, and to compare its characteristics to those of patients with idiopathic scoliosis from the same geographic area. Twenty-four [13 male and 11 female thalassemic patients aged 16 +/- 7 years (range 7-32 years)] of 115 examined patients with beta-thalassemia showed scoliosis of 14 degrees +/- 11 (range 10-65 degrees) radiologically. The prevalence of scoliosis in the thalassemic population was 21% in this series, whereas the overall prevalence of scoliosis in the general Greek population was 6% (Smyrnis PN, Valavanis J, Alexopoulos A, Siderakis G, Giannestras NJ: School screening for scoliosis in Athens, J Bone Joint Surg 61B:215-217, 1979). The scoliosis prevalence in the general population was significantly higher in the females (5%) than in the males (1%), whereas no difference in prevalence was found between the two sexes in the thalassemic population. The most common curve pattern in thalassemia was the left lumbar (38%) followed by the right lumbar (21%), whereas in patients with idiopathic scoliosis the left thoracolumbar most commonly appeared (25%) followed by the left lumbar (14%). No patient with thalassemia showed radiographic signs of congenital spinal deformities and spinal fractures, whereas all patients showed a significant retardation of their skeletal maturation. The age of the thalassemic patients with scoliosis was significantly (p = 0.0003) higher than in patients without scoliosis. The hematocrit of the thalassemic patients with scoliosis was significantly (p = 0.0012) lower than in those without scoliosis, whereas the rate of transfusions was not correlated with the magnitude of the scoliosis. The level of ferritin was significantly (p = 0.025) higher in the thalassemic patients with scoliosis than in those without scoliosis. The duration of Desferal treatment was significantly (p = 0.0357) longer in thalassemic patients with scoliosis when compared with those without scoliosis. Thus, the prevalence, curve pattern, and etiology of scoliosis in beta-thalassemia differ from those of idiopathic scoliosis, indicating that the spinal deformities in thalassemia represent a distinct type of scoliosis.


Examining the multidimensional role of self-efficacy for writing on student writing self-regulation and grades in elementary and high school.
The powerful role of self-efficacy on student achievement is clear. Less clear, however, is our understanding of the specific roles of the different factors of writing self-efficacy on student writing self-regulation and success. This study expands our knowledge about student writing self-efficacy - including its dimensions and the relationship between its dimensions and practical writing outcomes with students from different age groups. Participants were 992 elementary and 518 high school students across 65 teachers and 6 schools in the United States. Data were collected via online questionnaires, one assessing student writing self-efficacy and the others teachers' reports of student writing self-regulation and writing/ELA grades. Confirmatory factor analysis (CFA) results demonstrated significantly improved model fit for a three-factor model of writing self-efficacy compared to a one-factor model. A measurement invariance analysis of the writing self-efficacy scale indicated scalar invariance between the elementary and secondary samples. Structural equation modelling (SEM) results indicated that the second self-efficacy factor, self-efficacy for writing conventions, was statistically significantly related to writing/ELA grades for elementary students (β = .33, p < .001) as well as for high school students (β = .27, p < .01). Conventions were also the only factor statistically significantly related to teacher-reported student writing self-regulation for both elementary students (β = .26, p < .01) and high school students (β = .37, p < .001). Self-efficacy for ideation and self-efficacy for self-regulation were not found to be significantly related to either student writing/ELA grades or teacher-reported self-regulation (all ps > .19). Results support the multidimensionality of writing self-efficacy. In addition, findings emphasize a meaningful link between students' beliefs in their ability to conform to the rules of writing and their writing self-regulation and success.


Two- and three-dimensional image reconstructions from stained and autoradiographed histological sections.
A complete system has been developed to utilize histological serial sections for two- and three-dimensional image reconstructions. Eighty to 120 sections are digitized using a personal computing system augmented with a imaging board and CCD camera. The image files are transmitted to a VAX computer for processing and image reconstruction, and the processed images are transmitted back to the personal computer for display and recording using a film recorder or PostScript printer. The software developed for the system allows serial sections to be placed into proper registration in a 256(3) array, 256 grey levels. Autoradiographs of the sections are obtained in the presence of appropriate standards which are used to recalibrate grey levels to represent linearly the radioactivity of each pixel in the sections and scale the values to allow maximum use of the grey scale. Starting from coronally sectioned material the system has been used to analyse and reconstruct rat nasal turbinates. In two dimensions horizontal and sagittal sections have been obtained while in three dimensions back-to-front and surface-rendered images have been constructed. Useful rendering of differential metabolic activity within an organ of complex geometry has been obtained, and there appears to be no reason why the system cannot be used for any material for which serial sectioning is appropriate.


An optimal, generalized sampling time of 30 +/- 6 h after double dosing in the mouse peripheral blood micronucleus test.
Double dosing and single sampling seems to be the simplest and most reliable method for detecting clastogens in the mouse peripheral blood micronucleus test. Optimal sampling times after double dosing are studied here. Eleven clastogens [water soluble: colchicine, cyclophosphamide, cytosine arabinoside, 5-fluoro-2'-deoxyuridine, 5-fluorouracil (5-FU), 6-mercaptopurine, methotrexate (MTX), mitomycin C; water insoluble: N-2-acetylaminofluorene, diethyl sulfate, 7,12-dimethyl-benz[a]anthracene (DMBA)] were given i.p. twice to MS/Ae mice and peripheral blood samples were collected at 6 h intervals starting 24 h after the second treatment. Samples collected at 30 +/- 6 h were nearly optimal for all 11 chemicals. When DMBA, 5-FU and MTX were given, elevated micronucleus frequencies tended to last longer relative to those induced by direct-acting chemicals. Since samples collected 30 +/- 6 h after the second treatment with these three chemicals showed almost the same micronucleus frequencies observed in later samples, the time 30 +/- 6 h could be applied for these long-acting chemicals. Micronucleated reticulocytes persist in the peripheral blood (PB) longer than micronucleated polychromatic erythrocytes are retained in the bone marrow, which thus provides a simple, effective and generalized protocol for the mouse short-term PB micronucleus test, namely double dosing and sampling 30 +/- 6 h after the second dose. When one sample time has to be selected, 30 h after the second treatment is recommended.


Enhanced electrocatalytic activity of Pt subnanoclusters on graphene nanosheet surface.
Graphene nanosheet (GNS) gives rise to an extraordinary modification to the properties of Pt cluster electrocatalysts supported on it. The Pt/GNS electrocatalyst revealed an unusually high activity for methanol oxidation reaction compared to Pt/carbon black catalyst. The Pt/GNS electrocatalyst also revealed quite a different characteristic for CO oxidation among the measured catalyst samples. It is found that Pt particles below 0.5 nm in size are formed on GNS, which would acquire the specific electronic structures of Pt, modifying its catalytic activities.


Fulminant multiple system atrophy in a young adult presenting as motor neuron disease.
A 34-year-old man demonstrated rapidly progressive motor neuron disease and, late in his 9-month clinical course, exhibited ophthalmoplegia and dysautonomic symptoms. Neuropathology showed spinal and bulbar motor neuron disease with severe involvement of extraocular motor nuclei, degeneration of spinal sympathetic and bulbar parasympathetic nuclei, striatonigral degeneration, and early olivopontocerebellar atrophy. This case underscores the diversity of multiple system atrophy and demonstrates an unusually rapid course in a young patient.


Insulin modulates STAT3 protein activation and gene transcription in hepatic cells.
Treatment of rat hepatoma cells with insulin attenuated the interleukin 6 (IL-6) stimulation of acute phase plasma protein genes. To identify the potential mechanism of this action, the influence of insulin on IL-6 signal transduction was determined. An insulin dose-dependent reduction in signal transducer and activator of transcription 3 (STAT3) gene transcription, mRNA accumulation, protein concentration, and IL-6-inducible DNA binding activity was detected. A reduction in the IL-6-activated sis-inducible element binding of STAT3 was observed within 4 h of insulin treatment, whereas a maximal 3-4-fold lower STAT protein concentration was measured after 8-24 h of insulin treatment. Insulin mediated a similar magnitude reduction in the amount of mRNA encoding the IL-6 receptor alpha subunit and IL-6 binding activity. These effects of insulin appear to contribute to the strongly suppressed transcriptional induction of the IL-6-responsive acute phase plasma protein genes.


The expression of the renin-angiotensin-aldosterone system in the skin and its effects on skin physiology and pathophysiology.
The local renin-angiotensin-aldosterone system (RAAS) is fully expressed in the human skin at the mRNA and protein level. Local RAAS is known to play a regulatory function in epidermal proliferation, wound healing, scarring, cutaneous heating adaptation, and aging. There are also some indications of its role in the regulation of hair growth and sebum secretion. Impaired wound healing, skin diseases associated with diabetes, cancer development, psoriasis, and scleroderma may be related to changes in skin RAAS activity. Extensive research has shown that RAAS-modulating drugs can affect the skin when applied orally or topically, creating new therapeutic approaches against dermatological diseases.


Correlation of CT, MR, and Histopathology in Incomplete Partition-II Cochlear Anomaly.
To correlate the computed tomography (CT) and magnetic resonance imaging (MR) findings in a patient with incomplete partition-type II (IP-II) anomaly with histopathology in a similar human temporal bone specimen. To discover the histologic correlate of a common finding on MR of an "apparent" interscalar septum (IS). A patient with sensorineural hearing loss and imaging findings characteristic of IP-II, and a patient with histopathologic IP-II anomaly. High-resolution CT, MR, and review of postmortem temporal bone histopathology. Correlation of temporal bone histopathology with CT and MR findings. Consistent findings of IP-II anomaly on CT, absence of the IS between the more distal turns and flattening of the interscalar ridge between the distal basal turn and the middle turn, were present. The signal void surrounding the cochlea on MR also demonstrated flattening of the interscalar ridge. However, a thin band-like area of low T2 signal was seen, which could be mistaken for an IS. Correlation with temporal bone histopathology revealed that the modiolus was foreshortened, and the spiral ganglion neuron dendritic processes continued toward the upper middle turn through the osseous spiral lamina, likely accounting for the MR finding. Correlation of CT, MR, and histopathology in IP-II shows an "apparent" segmentation representing a continuum of neurosensory elements in approximately the same location of the expected location of a normal IS. Care should be taken when interpreting MR imaging in isolation. Understanding the bony outline of the cochlea on imaging may prove to be complementary.


An NMR, CD, molecular dynamics, and fluorometric study of the conformation of the bradykinin antagonist B-9340 in water and in aqueous micellar solutions.
A detailed NMR, CD, fluorometry, and molecular modeling study of a novel bradykinin antagonist B-9340, containing a novel amino acid D-Igl (alpha-(2-indanyl)glycine) at position 7, was carried out. The sequence of B-9340 is D-Arg0-Arg1-Pro2-Hyp3-Gly4-Thi5-Ser6-D- Igl7-Oic8-Arg9, where Hyp is hydroxyproline, Thi is beta-(2-thienyl)alanine, and Oic is (3aS,7aS)-octahydroindole-2-carboxylic acid. The CD results exhibit a striking effect of SDS on the spectrum of the BK antagonist, indicating that interaction with the surfactant induces a folded peptide structure. The interaction of this antagonist with phosphatidylinositol was monitored by fluorometry, indicating that the interaction of the peptide with the lipid is cooperative, and gives a Hill coefficient of 2.3. The two-dimensional proton NMR measurements indicate that B-9340 has no stable secondary structure in water solution and contains about 10-15% cis peptide bonds arising from Pro2, Hyp3, and Oic8. In SDS micelles, NMR reveals the existence of two beta-turns based on a number of medium-range connectivities that were useful for molecular modeling. The actual molecular modeling and dynamic runs were performed on B-9340 in an environment consisting of a layer of octyl sulfate anions and water. Ther results indicate that the structure of B-9340 in a micellar environment is characterized by a nonideal betaII-turn comprising residues Pro2 to Thi5, a nonideal betaII'-turn comprising residues Ser6-Arg9, and broad folding in the middle part of the molecule. The structure is stabilized by several hydrogen bonds and by a salt bridge between the guanidine moiety of Arg1 and the carboxyl group of Arg9, whereas the middle part of the peptide is buried in the micelle. The structure is deposited as Brookhaven PDB file 1 BDK.


Bismuth-Based Quadruple Therapy versus Metronidazole-Intensified Triple Therapy as a First-Line Treatment for Clarithromycin-Resistant Helicobacter pylori Infection: A Multicenter Randomized Controlled Trial.
Clarithromycin resistance is a main factor for treatment failure in the context of Helicobacter pylori infection. However, the treatment regimen for clarithromycin-resistant H. pylori infection has not yet been determined. We aimed to compare the efficacy and cost-effectiveness of 14-day bismuth-based quadruple therapy versus 14-day metronidazole-intensified triple therapy for clarithromycin-resistant H. pylori infection with genotypic resistance. This was a multicenter, randomized, controlled trial. A total of 782 patients with H. pylori infection examined using sequencing-based clarithromycin resistance point mutation tests were recruited between December 2018 and October 2020 in four institutions in Korea. Patients with significant point mutations (A2142G, A2142C, A2143G, A2143C, and A2144G) were randomly assigned to receive either 14-day bismuth-based quadruple therapy (n=102) or 14-day metronidazole-intensified triple therapy (n=99). The overall genotypic clarithromycin resistance rate was 25.7% according to the sequencing method. The eradication rate of 14-day bismuth-based quadruple therapy was not significantly different in the intention-to-treat analysis (80.4% vs 69.7%, p=0.079), but was significantly higher than that of 14-day metronidazole-intensified triple therapy in the per-protocol analysis (95.1% vs 76.4%, p=0.001). There were no significant differences in the incidence of side effects. In addition, the 14-day bismuth-based quadruple therapy was more cost-effective than the 14-day metronidazole-intensified triple therapy. Fourteen-day bismuth-based quadruple therapy showed comparable efficacy with 14-day metronidazole-intensified triple therapy, and it was more cost-effective in the context of clarithromycin-resistant H. pylori infection.


Prevalence of diabetes mellitus in the elderly of Namwon County, South Korea.
Ethnic and geographic differences exist in the prevalence of diabetes mellitus which has increased dramatically in South Korea. A few community-based epidemiologic studies with oral glucose tolerance test were performed in South Korea. The purpose of this study was to determine the prevalence of diabetes mellitus by the World Health Organization (WHO) and the American Diabetic Association (ADA) diagnostic criteria and to investigate their associated risk factors. Also, we compared and analyzed the characteristics of Koreans by WHO and ADA diagnostic criteria. Between March 22, 1999 and July 14, 1999, a random sampling of 1,445 residents over 40 years of age in five villages (3 myons and 2 dongs) in Namwon City. Chollabuk-do Province, South Korea was carried out. WHO and ADA diagnostic criteria were used for the prevalence of DM, IGT and IFG. The associated factors of subjects were analyzed. After age adjustment for the population projection of Korea (1999), the prevalence of DM and IGT was 13.7% and 13.8%, respectively, by WHO criteria, while the prevalence of DM, IGT and IFG was 15.8%, 12.8% and 5.7%, respectively, by ADA criteria, and the previous diagnosed diabetics were 5.8% in 665 adults over 40 years of age in the Namwon area. The age-adjusted prevalence of previously diagnosed diabetics was 5.8%. When the subjects classified by both criteria were compared, the level of agreement between WHO and ADA diagnostic criteria, except IFG, was very high (kappa = 0.94; p < 0.001). The ROC curve analysis determined FSG of 114.5 mg/dL (6.4 mmol/L) to yield optimal sensitivity and specificity corresponding to a PP2SG 200 mg/dL (11.1 mmol/L). The prevalence of DM and IGT with ADA diagnostic criteria rose with increasing age (p < 0.05). The body mass index was mean 23.8 +/- 3.4 in all the subjects, 23.75 +/- 3.46 in NGT group and 23.67 +/- 3.16 in DM group, but the differences in the prevalence of DM, IGT and IFG by BMI were not significant. The prevalence of DM rose significantly with the increase in the waist-hip ratio (p < 0.05). The prevalence of DM significantly increased in subjects by increases in blood pressure, and triglyceride and the relative risk in the prevalence of DM was significantly high with dyslipidemia (Odds ratio 2.29, 95% CI: 1.16-3.49). The prevalence of Diabetes Mellitus in the population over 40 years of age in Namwon City. South Korea remarkably increased compared with the 1970s and 1980s and was similar to that of the West. Ethnic differences in obesity of normal, DM and IGT subjects and in the effect on the prevalence of DM may exist in the Korean population, but they were not significant. As there is a limit in number, it is considered that a general population-based epidemiologic study on a large scale is required to investigate ethnic and geographic differences for the risk factors of DM in South Korea. The level of agreement, except IFG, by WHO and ADA diagnostic criteria was high, which indicates that these results may show that not only fasting serum glucose but also postprandial 2-h serum glucose are important for diagnosing diabetes in Korean.


Screening of carbohydrates in urine by capillary electrophoresis.
Carbohydrate analysis of urine is clinically important in assisting diagnosis of disorders of carbohydrate metabolism and understanding its pathologic significance. Paper chromatography and thin-layer chromatography are the techniques that are often employed for the determination of urinary carbohydrates. An aim of our experiments was to investigate the utility of capillary electrophoresis to develop a fast screening procedure of urinary carbohydrates. Simultaneous resolution of eight carbohydrates involving maltose, lactose, D-mannose, D-glucose, D-ribose, D-xylose, L-arabinose and D-galactose as markers was obtained with 130 mM borate (pH 10.2). Ethanol/water (80/20 v/v) and acetonitrile proved to be efficient reagents for urine sample clean-up which produced symmetrical peaks. The urine sample from a normal subject was determined to contain lactose, glucose, xylose and arabinose that fall within normal ranges of these carbohydrates in urine. The investigations made in this study may be potentially useful in carbohydrate screening, especially in neonatal urine screening.


Genomic instability in squamous cell carcinoma of the head and neck.
The role of genomic instability in the development of squamous cell carcinoma (SCCHN) has become apparent with the publication of three major allelotype analysis of this disease, as well as many publications which have concentrated on specific target regions. The measurement of accumulated genetic alterations or fractional allele loss, as determined by allelotype analysis, provides a useful molecular indicator of tumour behaviour. In one major study, a positive correlation was found between FAL > median value and lymph node metastasis and also with a poor clinical outcome. In addition the recognition of microsatellite instability as a marker of DNA repair defects has provided a further molecular marker of the disease process and that loss of heterozygosity analysis and microsatellite instability appear to be independent genetic events in the development of SCCHN. Furthermore, the recognition of a number of novel target regions in SCCHN on chromosome arms, 1 p, 3p, 8p, 9p, 13q, 17p and 18q and our understanding of the role of certain oncogenes and tumour suppressor genes and their interaction with human papillomavirus has provided further elucidation of the neoplastic process. Even though this review describes a number of molecular events in SCCHN, the sequence of events still eludes the scientific community at present.


Effects of loading on maximum vertical jumps: Selective effects of weight and inertia.
A novel loading method was applied to explore selective effects of externally added weight (W), weight and inertia (W+I), and inertia (I) on maximum counter-movement jumps (CMJ) performed with arm swing. Externally applied extended rubber bands and/or loaded vest added W, W+I, and I corresponding to 10-40% of subjects' body mass. As expected, an increase in magnitude of all types of load was associated with an increase in ground reaction forces (GRF), as well as with a decrease in both the jumping performance and power output. However, of more importance could be that discernible differences among the effects of W, W+I, and I were recorded despite a relatively narrow loading range. In particular, an increase in W was associated with the minimal changes in movement kinematic pattern and smallest reduction of jumping performance, while also allowing for the highest power output. Conversely, W+I was associated with the highest ground reaction forces. Finally, the lowest maxima of GRF and power were associated with I. Although further research is apparently needed, the obtained finding could be of potential importance not only for understanding fundamental properties of the neuromuscular system, but also for optimization of loading in standard athletic training and rehabilitation procedures.


Effect of Salicylic Acid Pre-Treatment after Long-Term Desiccation in the Moss Syntrichia ruralis (Hedw.) Web. and Mohr.
The main objective of this research was to examine the effects of exogenous salicylic acid (SA) and to study the seasonal variation of the chlorophyll a fluorescence parameters and antioxidant enzymatic activities in desiccation-tolerant moss species Syntrichia ruralis (Hedw.) Web. and Mohr. Aqueous 0.001 M SA solution was sprayed on the moss cushions collected from semi-arid sandy grassland, Hungary in three seasons (spring, summer, autumn). These cushions were kept under the observation for 10 Days. Chlorophyll a fluorescence parameters, i.e., maximum photochemical quantum yield of PS II (Fv/Fm), effective photochemical quantum yield of PS II (ΦPSII), non-photochemical quenching (NPQ), and antioxidant enzymatic activities, i.e., ascorbate peroxidase (APX), catalase (CAT), guaiacol peroxidase (POD), and protein content were determined. The results showed the increase of Fv/Fm in spring and autumn season while ΦPSII was reduced significantly during spring and summer season after treatment with SA compared to control. SA-treated mosses showed higher values of non-photochemical quenching (NPQ) during the spring and autumn season than in summer. Activities of enzyme APX and CAT were found to increase in SA-treated except POD activity. In SA-treated moss cushions, lower protein content was found. It can be concluded that seasonal variation has been observed in chlorophyll fluorescence and antioxidant system after long term of desiccation in S. ruralis species that could be because of SA and might be due to fluctuations in conditions of their habitat, duration of light intensity, temperature and precipitation.


An integrative multi-omics analysis based on liquid-liquid phase separation delineates distinct subtypes of lower-grade glioma and identifies a prognostic signature.
Emerging evidences have indicated that the aberrant liquid-liquid phase separation (LLPS) leads to the dysfunction of biomolecular condensates, thereby contributing to the tumorigenesis and progression. Nevertheless, it remains unclear whether or how the LLPS of specific molecules affects the prognosis and tumor immune microenvironment (TIME) of patients with lower-grade glioma (LGG). We integrated the transcriptome information of 3585 LLPS-related genes to comprehensively evaluate the LLPS patterns of 423 patients with LGG in The Cancer Genome Atlas (TCGA) cohort. Then, we systematically demonstrated the differences among four LLPS subtypes based on multi-omics analyses. In addition, we constructed the LLPS-related prognostic risk score (LPRS) for individualized integrative assessment. Based on the expression profiles of 85 scaffolds, 355 regulators, and 3145 clients in LGG, we identified four LLPS subtypes, namely LS1, LS2, LS3 and LS4. We confirmed that there were significant differences in prognosis, clinicopathological features, cancer hallmarks, genomic alterations, TIME patterns and immunotherapeutic responses among four LLPS subtypes. In addition, a prognostic signature called LPRS was constructed for individualized integrative assessment. LPRS exhibited a robust predictive capacity for prognosis of LGG patients in multiple cohorts. Moreover, LPRS was found to be correlated with clinicopathological features, cancer hallmarks, genomic alterations and TIME patterns of LGG patients. The predictive power of LPRS in response to immune checkpoint inhibitor (ICI) therapy was also prominent. This study provided a novel classification of LGG patients based on LLPS. The constructed LPRS might facilitate individualized prognosis prediction and better immunotherapy options for LGG patients.


Catalase Degradation in Sunflower Cotyledons during Peroxisome Transition from Glyoxysomal to Leaf Peroxisomal Function.
First order rate constants for the degradation (degradation constants) of catalase in the cotyledons of sunflower (Helianthus annuus L.) were determined by measuring the loss of catalase containing (14)C-labeled heme. During greening of the cotyledons, a period when peroxisomes change from glyoxysomal to leaf peroxisomal function, the degradation of glyoxysomal catalase is significantly (P = 0.05) slower than during all other stages of cotyledon development in light or darkness. The degradation constant during the transition stage of peroxisome function amounts to 0.205 day(-1) in contrast to the constants ranging from 0.304 day(-1) to 0.515 day(-1) during the other developmental stages. Density labeling experiments comprising labeling of catalase with (2)H(2)O and its isopycnic centrifugation on CsCl gradients demonstrated that the determinations of the degradation constants were not substantially affected by reutilization of (14)C-labeled compounds for catalase synthesis. The degradation constants for both glyoxysomal catalase and catalase synthesized during the transition of peroxisome function do not differ. This was shown by labeling the catalases with different isotopes and measuring the isotope ratio during the development of the cotyledons. The results are inconsistent with the concept that an accelerated and selective degradation of glyoxysomes underlies the change in peroxisome function. The data suggest that catalase degradation is at least partially due to an individual turnover of catalase and does not only result from a turnover of the whole peroxisomes.


Rapid characterization of N-linked glycosylation site using non-specific protease digestion of gel-separated glycoproteins and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry.
Rapid identification of glycosylation sites of glycoproteins is urgently needed in glycoproteomics study. In the present work, a rapid and simple method based on non-specific digestion of gel-separated glycoproteins and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry was described, which can efficiently identify the N-linked glycosylation sites. One-step in-gel digestion of Ribonuclease B (RNase B) by proteinase K was employed to generate glycopeptides with short and discrepant peptide composition. When compared with glycopeptides prepared by two-step in gel-digestion using trypsin-proteinase K or trypsin-pronase, the direct proteinase K treatment showed obvious superiority in both glycopeptide recovery and preparation simplicity. Most importantly, it helps to generate greater variety of glycopeptide series with rich information for glycosylation site identification. In addition, binary matrices 5-chloro-2-mercaptobenzothiazole (CMBT) /2,5-dihydroxybenzoic acid (DHB) were found to form homogeneous microcrystal on the target with the purified glycopeptides, leading to improved detection sensitivity. Thus, the present work provides an optimized solution to speed up the characterization of N-linked glycosylation sites in glycoproteins.


Application of LC-MS/MS MRM to Determine Staphylococcal Enterotoxins (SEB and SEA) in Milk.
Staphylococcus aureus is one of the important aetiological agents of food intoxications in Europe and can cause gastro-enteritis through the production of various staphylococcal enterotoxins (SEs) in foods. Due to their stability and ease of production and dissemination, some SEs have also been studied as potential agents for bioterrorism. Therefore, specific and accurate analytical tools are required to detect and quantify SEs. Online solid-phase extraction liquid chromatography electrospray ionization tandem mass spectrometry (online SPE-LC-ESI-MS/MS) based on multiple reaction monitoring (MRM) was used to detect and quantify two types of SE (A and B) spiked in milk and buffer solution. SE extraction and concentration was performed according to the European Screening Method developed by the European Reference Laboratory for Coagulase Positive Staphylococci. Trypsin digests were screened for the presence of SEs using selected proteotypic heavy-labeled peptides as internal standards. SEA and SEB were successfully detected in milk samples using LC-MS/MS in MRM mode. The selected SE peptides were proteotypic for each toxin, allowing the discrimination of SEA and SEB in a single run. The detection limit of SEA and SEB was approximately 8 and 4 ng/g, respectively.


Differences in CD8αα and cecal microbiome community during proliferation and late cytolytic phases of Marek's disease virus infection are associated with genetic resistance to Marek's disease.
Marek's disease (MD) is an important neoplastic disease of chickens caused by Marek ': s disease virus (MDV), a highly oncogenic alphaherpesvirus. In this study using two chicken lines, one resistant and another susceptible to MD, splenic T cells and cecal microbiome were profiled to gain a better understanding of primary differences in these lines. The percent of splenic CD4+ T cells were similar regardless of MDV challenge status in both bird lines. In contrast, CD8αα profiles were different (P < 0.005) between chicken lines under naïve status and under MDV challenge, suggesting that CD8αα T cells play a key role in mediating MDV infection. Microbiome composition was different between naïve resistant (Blautia spp.) and susceptible birds (Streptococcus spp.) (P < 0.05) during initial colonization. With MDV challenge, both chicken lines showed lower numbers of beneficial Faecalibacterium spp. and increased number of Lactobacillus spp. Metabolic profiles between naïve chicken types were similar but with MDV challenge, there were differences in metabolism in both chicken lines, with amino acid metabolism impacted in resistant birds and lipid metabolism in susceptible birds. These results provide insights into immune response and potential interplay with the microbiome during infection with an oncogenic virus.


Applications of the Heligman/Pollard model mortality schedule.
"The United Nations working manual for MORTPAK-Lite, a software package for demographic measurement, includes among its 16 computer programs a routine (UNABR) that graduates a set of age-specific probabilities of dying...for the standard set of five-year age groups into a set of single-year probabilities of dying. The graduation is effected with an eight-parameter 'law of mortality' known as the Heligman/Pollard model mortality schedule. Model mortality schedules...are useful in mortality analysis and forecasting. Several such applications are illustrated in this article: a historical time-series analysis, a forecasting application, studies of causes of death, spatial differences and sex-specific mortality disaggregated by race." The geographical focus is on the United States, England, and Australia.


Impact of the Breakpoint Region on the Leukemogenic Potential and the TKI Responsiveness of Atypical BCR-ABL1 Transcripts.
Chronic Myeloid Leukemia (CML) is a hematological disorder characterized by the clonal expansion of a hematopoietic stem cell carrying the Philadelphia chromosome that juxtaposes the BCR and ABL1 genes. The ensuing BCR-ABL1 chimeric oncogene is characterized by a breakpoint region that generally involves exons 1, 13 or 14 in BCR and exon 2 in ABL1. Additional breakpoint regions, generating uncommon BCR-ABL1 fusion transcripts, have been detected in various CML patients. However, to date, the impact of these infrequent transcripts on BCR-ABL1-dependent leukemogenesis and sensitivity to tyrosine kinase inhibitors (TKIs) remain unclear. We analyzed the transforming potential and TKIs responsiveness of three atypical BCR-ABL1 fusions identified in CML patients, and of two additional BCR-ABL1 constructs with lab-engineered breakpoints. We observed that modifications in the DC2 domain of BCR and SH3 region of ABL1 affect BCR-ABL1 catalytic efficiency and leukemogenic ability. Moreover, employing immortalized cell lines and primary CD34-positive progenitors, we demonstrate that these modifications lead to reduced BCR-ABL1 sensitivity to imatinib, dasatinib and ponatinib but not nilotinib. We conclude that BCR-ABL1 oncoproteins displaying uncommon breakpoints involving the DC2 and SH3 domains are successfully inhibited by nilotinib treatment.


Bilateral fungal keratitis with ring infiltrates: a rare scenario.
A 12-year-old boy presented to the emergency department with chief complaints of pain, redness, discharge and diminution of vision in both eyes over the previous 20 days. There was no history of preceding trauma, contact lens use, any eye drop usage or ocular surgery. Systemic history was not significant. Presenting uncorrected visual acuity in his right eye was counting fingers at 1 m and 20/200 in the left eye, with accurate projection of rays in both eyes. Slit lamp biomicroscopy showed the presence of bilateral diffuse conjunctival congestion, corneal ring infiltrates and epithelial defect with corneal oedema. Potassium hydroxide wet mount showed the presence of septate fungal hyphae. The patient was treated with topical 5% natamycin and 1% voriconazole over a period of 6 weeks. Best-corrected visual acuity was 20/600 in the right eye and 20/20 in the left eye at 6-month follow-up.


Impact of Telemedicine on Severity of Illness and Outcomes Among Children Transferred From Referring Emergency Departments to a Children's Hospital PICU.
To compare the severity of illness and outcomes among children admitted to a children's hospital PICU from referring emergency departments with and without access to a pediatric critical care telemedicine program. Retrospective cohort study. Tertiary academic children's hospital PICU. Pediatric patients admitted directly to the PICU from referring emergency departments between 2010 and 2014. None. Demographic factors, severity of illness, and clinical outcomes among children receiving care in emergency departments with and without access to pediatric telemedicine, as well as a subcohort of children admitted from emergency departments before and after the implementation of telemedicine. Five hundred eighty-two patients from 15 emergency departments with telemedicine and 524 patients from 60 emergency departments without telemedicine were transferred and admitted to the PICU. Children admitted from emergency departments using telemedicine were younger (5.6 vs 6.9 yr; p< 0.001) and less sick (Pediatric Risk of Mortality III score, 3.2 vs 4.0; p < 0.05) at admission to the PICU compared with children admitted from emergency departments without telemedicine. Among transfers from emergency departments that established telemedicine programs during the study period, children arrived significantly less sick (mean Pediatric Risk of Mortality III scores, 1.2 units lower; p = 0.03) after the implementation of telemedicine (n = 43) than before the implementation of telemedicine (n = 95). The observed-to-expected mortality ratios of posttelemedicine, pretelemedicine, and no-telemedicine cohorts were 0.81 (95% CI, 0.53-1.09), 1.07 (95% CI, 0.53-1.60), and 1.02 (95% CI, 0.71-1.33), respectively. The implementation of a telemedicine program designed to assist in the care of seriously ill children receiving care in referring emergency departments was associated with lower illness severity at admission to the PICU. This study contributes to the body of evidence that pediatric critical care telemedicine programs assist referring emergency departments in the care of critically ill children and could result in improved clinical outcomes.


A novel method for simultaneous and continuous determination of thermal properties during phase transition applied to Calanus finmarchicus.
The thermal properties of a product are the most important parameters for practical engineering purposes and models in food science. Calanus finmarchicus is currently being examined as a marine resource for uncommon aquatic lipids and proteins. Thermal conductivity, specific heat, enthalpy and density were measured over the temperature range from -40 to +20 degrees C. The initial freezing point was determined to be -2.3 degrees C. The thermal properties were recorded continuously on 4 samples using a new method, and the results were compared with predictive models. The accuracy of the new method is demonstrated by different calibration runs. Significant differences in the thermal conductivity of the frozen material were found between the parallel-series model and the data, whereas the model of Pham and Willix (1989) or the Maxwell-Euken adaption showed better agreement. The measured data for specific heat, enthalpy, and density agreed well with the model. Practical Application: The thermal data obtained can be used directly in food engineering and technology applications, for example, in a thin layer model for freezing food for which precise thermal data for each layer are now available, enabling the more accurate prediction of freezing times and temperature profiles. Dimensionless numbers (such as the Biot number) can also be based on measured data with minor deviations compared to more general modeled thermal properties. Future activities will include the generation of a comprehensive database for different products.


The role of Fe(III) on phosphate released during the photo-decomposition of organic phosphorus in deionized and natural waters.
The photo-decomposition of organic phosphorus is an important route for the phosphorus cycle by which phosphate is regenerated in the aquatic environment. In this study, the role of Fe3+ as a natural photosensitizer toward the decomposition of organic phosphorus to release phosphate was examined in deionized and natural waters under UV and sunlight irradiation using glyphosate as the organic phosphorus model. The results showed that the concentration of glyphosate decreased with irradiation time in the Fe3+/UV and Fe3+/sunlight systems and TOC gradually decreased, which confirmed that glyphosate was degraded by Fe3+. The amount of phosphate released from the photo-decomposition of glyphosate was higher in the presence of Fe3+ than that of the control experiment under UV and sunlight irradiation conditions, and the generation rate of phosphate also increased with increasing Fe3+concentrations. The formation of hydroxyl radicals (·OH) in the Fe3+/UV and Fe3+/sunlight systems was identified according to the photoluminescence spectra (PL) using coumarin as the trapping molecule, and the steady-state concentrations of ·OH for the Fe3+/UV and Fe3+/sunlight systems were 1.06 × 10-14 M and 0.09 × 10-14 M, respectively. When natural water was spiked with glyphosate and Fe3+, the phosphate that was released in the Fe3+ was higher than that of the control, and the phosphate that was released was inhibited when isopropanol was added to the reaction. All of these results demonstrate that the photochemical activity of Fe3+ has significantly impact in the release of phosphate from the photo-decomposition of organic phosphorus.


Liposomal Bupivacaine (Bupigel) Demonstrates Minimal Local Nerve Toxicity in a Rabbit Functional Model.
We previously reported the development of a novel formulation of an ultra-long-acting local anesthetic based on bupivacaine encapsulated in large multivesicular liposomes (Bupisomes) embedded in hydrogel. This formulation (Bupigel) prolonged bupivacaine release from the formulation in dissolution-like studies in vitro and analgesia in vivo in mouse, rat, and pig models. In this study we assessed Bupigel neurotoxicity on rabbit sciatic nerve using histopathology and electrophysiologic testing. Sciatic nerves of both hind limbs were injected dropwise with different formulations. Nerve conduction studies and needle electromyography two weeks after perineural administration showed signs of neural damage after injection of free lidocaine and bupivacaine, while there was no sign of neural damage after injection with saline, demonstrating the validity of the method. This test also did not show evidence of motor or sensory nerve damage after injection with liposomal bupivacaine at a dose 10-times higher than free bupivacaine. Histologically, signs of neural damage could be observed with lidocaine. Nerves injected with Bupigel showed mild signs of inflammation and small residues of hydrogel in granulomas, indicating a long residence time of the hydrogel at the site of injection, but no histopathological signs of nerve damage. This demonstrated that early signs of neural damage were detected electrophysiologically, showing the usefulness and sensitivity of electrodiagnostic testing in detection of neural damage from new formulations.


Discordance in Tumor Mutation Burden from Blood and Tissue Affects Association with Response to Immune Checkpoint Inhibition in Real-World Settings.
Tumor mutation burden (TMB), a biomarker for immune checkpoint inhibitor (CPI) response, is reported by both blood- and tissue-based next-generation sequencing (NGS) vendors. However, the agreement between TMB from blood (bTMB) and tissue (tTMB) in real-world settings, both in absolute value and association with CPI response, is not known. This study utilizes Sarah Cannon's precision medicine platform, Genospace, to harmonize clinico-genomic data from 17 206 patients with cancer with NGS results from September 2015 to August 2021. A subset of patients have both bTMB and tTMB results. Statistical analyses are performed in R and include (1) correlation (r) and concordance (ρ) between patient-matched bTMB-tTMB pairs, (2) distribution of total bTMB and tTMB values, and (3) association of bTMB and tTMB with time to CPI therapy failure. In 410 patient-matched bTMB-tTMB pairs, the median bTMB (m = 10.5 mut/Mb) was significantly higher than the median tTMB (m = 6.0 mut/Mb, P < .001) leading to conflicting "high" and "low" statuses in over one-third of cases at a threshold of 10 mut/Mb (n = 410). Significant differences were observed in the distribution of bTMB values from blood-NGS vendors, with guardant health (GH) reporting higher (m = 10.5 mut/Mb, n = 2183) than Foundation Medicine (FMI, m = 3.8 mut/Mb, n = 462, P < .001). bTMB from GH required a higher threshold (≥40 mut/Mb) than bTMB from FMI (≥12 mut/Mb) in order to be associated with CPI response. This study uncovers variability in bTMB reporting among commercial NGS platforms, thereby evidencing a need for assay-specific thresholds in identifying patients who may respond to CPI therapy.


Light Emission Enhancement by Tuning the Structural Phase of APbBr3 (A = CH3NH3, Cs) Perovskites.
Lead halide perovskite (APbX3) has recently emerged as a promising active layer in light-emitting diodes (LEDs) as well as an absorber for photovoltaic devices. For better LED properties, it is important to understand the fundamental mechanism of the optoelectronic behaviors, e.g., how the nanostructure of the APbX3 thin film correlates with its emitting properties. We investigated the effect of APbBr3 (A = CH3NH3, Cs) crystallite size on the photophysical properties regarding its crystallographic changes and spin-orbit coupling. Photoluminescence lifetime measurements, X-ray and electron diffraction analyses, and density functional theory calculations were performed. We demonstrate that the emitting properties of mesoscale APbBr3 crystallites are improved due to the formation of a pure cubic phase that leads to the spin- and momentum-allowed carrier recombination. Our findings provide fundamental insights into the emitting behavior of APbBr3, which suggests a control of its optoelectronic properties by means of modulating the crystal morphology and resultant electronic band structures.


Agent-based modeling: case study in cleavage furrow models.
The number of studies in cell biology in which quantitative models accompany experiments has been growing steadily. Roughly, mathematical and computational techniques of these models can be classified as "differential equation based" (DE) or "agent based" (AB). Recently AB models have started to outnumber DE models, but understanding of AB philosophy and methodology is much less widespread than familiarity with DE techniques. Here we use the history of modeling a fundamental biological problem-positioning of the cleavage furrow in dividing cells-to explain how and why DE and AB models are used. We discuss differences, advantages, and shortcomings of these two approaches.


Tailoring the Structural and Optical Properties of Germanium Telluride Phase-Change Materials by Indium Incorporation.
Chalcogenide phase-change materials (PCMs) based random access memory (PCRAM) enter the global memory market as storage-class memory (SCM), holding great promise for future neuro-inspired computing and non-volatile photonic applications. The thermal stability of the amorphous phase of PCMs is a demanding property requiring further improvement. In this work, we focus on indium, an alloying ingredient extensively exploited in PCMs. Starting from the prototype GeTe alloy, we incorporated indium to form three typical compositions along the InTe-GeTe tie line: InGe3Te4, InGeTe2 and In3GeTe4. The evolution of structural details, and the optical properties of the three In-Ge-Te alloys in amorphous and crystalline form, was thoroughly analyzed via ab initio calculations. This study proposes a chemical composition possessing both improved thermal stability and sizable optical contrast for PCM-based non-volatile photonic applications.


A case of Crohn's disease with hydronephrosis caused by ureteropelvic junction obstruction.
Ureteral obstruction is a rare extraintestinal manifestation of Crohn's disease (CD). We report the case of a 20-year-old man who presented with fever, diarrhoea, and lower abdominal pain. Diagnostic studies confirmed CD and revealed an abdominal mass obstructing the right ureter and hydronephrosis. Ureteropelvic junction (UPJ) obstruction was diagnosed. Despite an elemental diet and mesalazine therapy, the hydronephrosis persisted, and the patient eventually required surgery. Successful laparoscopic pyeloplasty was performed. This is the first report of CD associated with UPJ obstruction. Ureteral obstruction as a complication of CD is discussed.


Localization of diacylglycerol lipase-alpha around postsynaptic spine suggests close proximity between production site of an endocannabinoid, 2-arachidonoyl-glycerol, and presynaptic cannabinoid CB1 receptor.
2-arachidonoyl-glycerol (2-AG) is an endocannabinoid that is released from postsynaptic neurons, acts retrogradely on presynaptic cannabinoid receptor CB1, and induces short- and long-term suppression of transmitter release. To understand the mechanisms of the 2-AG-mediated retrograde modulation, we investigated subcellular localization of a major 2-AG biosynthetic enzyme, diacylglycerol lipase-alpha (DAGLalpha), by using immunofluorescence and immunoelectron microscopy in the mouse brain. In the cerebellum, DAGLalpha was predominantly expressed in Purkinje cells. DAGLalpha was detected on the dendritic surface and occasionally on the somatic surface, with a distal-to-proximal gradient from spiny branchlets toward somata. DAGLalpha was highly concentrated at the base of spine neck and also accumulated with much lower density on somatodendritic membrane around the spine neck. However, DAGLalpha was excluded from the main body of spine neck and head. In hippocampal pyramidal cells, DAGLalpha was also accumulated in spines. In contrast to the distribution in Purkinje cells, DAGLalpha was distributed in the spine head, neck, or both, whereas somatodendritic membrane was labeled very weakly. These results indicate that DAGLalpha is essentially targeted to postsynaptic spines in cerebellar and hippocampal neurons, but its fine distribution within and around spines is differently regulated between the two neurons. The preferential spine targeting should enable efficient 2-AG production on excitatory synaptic activity and its swift retrograde modulation onto nearby presynaptic terminals expressing CB1. Furthermore, different fine localization within and around spines suggests that the distance between postsynaptic 2-AG production site and presynaptic CB1 is differentially controlled depending on neuron types.


Avian Hepatitis E Virus ORF2 Protein Interacts with Rap1b to Induce Cytoskeleton Rearrangement That Facilitates Virus Internalization.
Avian hepatitis E virus (HEV) causes liver diseases and multiple extrahepatic disorders in chickens. However, the mechanisms involved in avian HEV entry remain elusive. Herein, we identified the RAS-related protein 1b (Rap1b) as a potential HEV-ORF2 protein interacting candidate. Experimental infection of chickens and cells with an avian HEV isolate from China (CaHEV) led to upregulated expression and activation of Rap1b both in vivo and in vitro. By using CaHEV capsid as mimic of virion to treat cell in vitro, it appears that the interaction between the viral capsid and Rap1b promoted cell membrane recruitment of the downstream effector Rap1-interacting molecule (RIAM). In turn, RIAM further enhanced Talin-1 membrane recruitment and retention, which led to the activation of integrin α5/β1, as well as integrin-associated membrane protein kinases, including focal adhesion kinase (FAK). Meanwhile, FAK activation triggered activation of downstream signaling molecules, such as Ras-related C3 botulinum toxin substrate 1 RAC1 cell division cycle 42 (CDC42), p21-activated kinase 1 (PAK1), and LIM domain kinase 1 (LIMK1). Finally, F-actin rearrangement induced by Cofilin led to the formation of lamellipodia, filopodia, and stress fibers, contributes to plasma membrane remodeling, and might enhance CaHEV virion internalization. In conclusion, our data suggested that Rap1b activation was triggered during CaHEV infection and appeared to require interaction between CaHEV-ORF2 and Rap1b, thereby further inducing membrane recruitment of Talin-1. Membrane-bound Talin-1 then activates key Integrin-FAK-Cofilin cascades involved in modulation of actin kinetics, and finally leads to F-actin rearrangement and membrane remodeling to potentially facilitate internalization of CaHEV virions into permissive cells. IMPORTANCE Rap1b is a multifunctional protein that is responsible for cell adhesion, growth, and differentiation. The inactive form of Rap1b is phosphorylated and distributed in the cytoplasm, while active Rap1b is prenylated and loaded with GTP to the cell membrane. In this study, the activation of Rap1b was induced during the early stage of avian HEV infection under the regulation of PKA and SmgGDS. Continuously activated Rap1b recruited its effector RIAM to the membrane, thereby inducing the membrane recruitment of Talin-1 that led to the activation of membrane α5/β1 integrins. The triggering of the signaling pathway-associated Integrin α5/β1-FAK-CDC42&RAC1-PAK1-LIMK1-Cofilin culminated in F-actin polymerization and membrane remodeling that might promote avian HEV virion internalization. These findings suggested a novel mechanism that is potentially utilized by avian HEV to invade susceptible cells.


Manipulation of the immune response by foreign gene expression in the thymus.
Retroviral gene transfer vectors have been developed for optimal in vivo gene therapy. Ideally, these vectors should target gene expression specifically to selected tissues or organs. Our studies focus on the development of retroviral vectors for gene delivery to the thymus. The goal of these studies is to utilize thymic expression of exogenous genes to manipulate the immune repertoire. We have characterized the selective thymic tropism of a molecular clone of Gross murine leukemia virus, GD-17, to thymic medullary epithelial cells using immunohistochemical staining and confocal microscopy. Specific expression of viral antigens in the thymus lead to the induction of immunologic tolerance to GMuLV proteins. This tissue specific vector may thus be used to study the requirements of epithelial mediated tolerance induction, and provide a more efficient tool for gene therapy.


Composition and Function of Thylakoid Membranes from Grana-rich and Grana-deficient Chloroplast Mutants of Barley.
Chlorophyll-deficient barley (Hordeum vulgare) mutants were studied that had chlorophyll a/b ratios either higher or lower than the wild type. Mutants with high ratios (>5.2) had a reduced proportion of their photosynthetic lamellae appressed into grana ("grana-deficient" mutants) compared with wild type (chlorophyll a/b = 3.2), while the majority of lamellae in the chloroplasts with low chlorophyll a/b ratios (2.0-2.4) were organized into grana ("grana-rich" mutants).All mutants catalyzed photosystem I and photosystem II electron transport, were tightly coupled as evidenced by increased rates of electron transport in the presence of methylamine, and were able to generate a light-dependent transmembrane proton gradient. Differences were evident in rates of electron transport per mole of chlorophyll. The mutants having high chlorophyll a/b ratios catalyzed 15- to 50-fold higher rates of ferricyanide photoreduction than the mutants having low chlorophyll a/b ratios, and 5- to 7-fold higher than the wild type.Low temperature absorption spectra of chloroplast fragments showed that the grana-deficient mutant with a high a/b ratio had a chlorophyll spectrum characteristic of a PSI preparation while mutants with the low ratio had a spectrum typical of a PSII preparation.The temperature fluorescence emission spectra of thylakoid membrane fragments from the two types of mutants were also strikingly different from one another, as were the electrophoretic patterns of the thylakoid polypeptides.


Three-dimensional metal azide coordination polymers with amino carboxylate coligands: synthesis, structure, and magnetic properties.
The reaction of M(II) ions with azido ligands in the presence of different amino carboxylic acids gave four three-dimensional metal-azido coordination polymers, [Mn(3,5-daba)(N(3))](n) (1), [Cd(3,5-daba)(N(3))](n) (2; 3,5-daba = 3,5-diaminobenzoate), [Mn(4-aba)(N(3))](n) (3; 4-aba = 4-aminobenzoate), and [Cu(2)(gly)(2)(N(3))(2)](n) (4; gly = glycinate), which display different topological structures. Polymers 1 and 2 present 4,6-connected 3D networks with different Schlafli symbols. However, 3 and 4 feature an unprecedented trinodal 3,6-connected network with the Schlafli symbol (4(2).6)(4.6(2))(4(3).6(6).8(6)) and an unusual 4-connected 3D net with the Lonsdaleite (hexagonal diamond) topology, respectively. Magnetic susceptibility measurements revealed dominant antiferromagnetic couplings for 1 and 3 and an overall dominant ferromagnetic coupling for 4, which presents metamagnetic behavior with a magnetic phase transition at a critical temperature of 6 K and a transition field of ca. 6030 Oe. The results demonstrate that the EE azido and syn-anti carboxylato bridges in our cases induce an antiferomagnetic interaction, and the anti-anti carboxylato bridge in 4 mediates a ferromagnetic interaction. The magnetic interaction through the EO azido bridge in 3 and 4 has a dependence on the value of the M-N-M bond angle.


The dominant-negative interplay between p53, p63 and p73: A family affair.
The tumor suppression activity of p53 is frequently impaired in cancers even when a wild-type copy of the gene is still present, suggesting that a dominant-negative effect is exerted by some of p53 mutants and isoforms. p63 and p73, which are related to p53, have also been reported to be subjected to a similar loss of function, suggesting that a dominant-negative interplay might happen between p53, p63 and p73. However, to which extent p53 hotspot mutants and isoforms of p53, p63 and p73 are able to interfere with the tumor suppressive activity of their siblings as well as the underlying mechanisms remain undeciphered. Using yeast, we showed that a dominant-negative effect is widely spread within the p53/p63/p73 family as all p53 loss-of-function hotspot mutants and several of the isoforms of p53 and p73 tested exhibit a dominant-negative potential. In addition, we found that this dominant-negative effect over p53 wild-type is based on tetramer poisoning through the formation of inactive hetero-tetramers and does not rely on a prion-like mechanism contrary to what has been previously suggested. We also showed that mutant p53-R175H gains the ability to inhibit p63 and p73 activity by a mechanism that is only partially based on tetramerization.


[Ciliary ablation with diode laser. Long-term study].
The effects of contact transscleral cyclophotocoagulation with diode laser for refractory glaucoma have been analyzed. A diode laser system was used to treat twenty one eyes with refractory glaucoma. The mean follow-up was 22.24+/-9.89 months. All the eyes were treated with 15 laser spots placed over 270 degrees, 1 mm posterior to the limbus, using 3.0 J (1.0 W X 3.0 sg). The mean preoperative intraocular pressure was 40.95+/-11.76 mmHg and the mean intraocular pressure at the end of the follow-up was 24.28+/-7.80 mmHg (p<0.5). An intraocular pressure below 21 mmHg was obtained in 10 out the 13 eyes (76.92%). Six of the 8 painful eyes had pain relief (75.00%). The principal complications included a decreased visual acuity in 5 eyes and chronic uveitis in 2 patients. These results suggest that semiconductor diode transscleral cyclophotocoagulation can be used successfully to reduce intraocular pressure in the treatment of refractory glaucoma.


Implementation of program of prevention and early detection of osteoporosis among women of Primorsko-goranska County.
The aim of this paper is to present preliminary data of Program of prevention and early detection of osteoporosis among women in Primorsko-goranska County. Osteoporosis is recognized as a public health problem for which clearly preventive measures are defined. Measurement of bone density was done by ultrasound densitometry of the calcaneus among women aged from 45 to 69 years old. 688 women were examined and they were classified in five five-year age groups. The women with the osteoporosis (T-score < or = 2.5) were 141; osteopenia (T-score from -2.5 to -1) were found in 400 women, and those with normal range of T-score were 147. All of five groups of women had T-score in range of osteopenia (T-score < or = 1). A statistically significant difference was between the first and fourth groups of women (p = 0.002) and the second and fourth groups (p = 0.001). After examination, depending on the value of T-score, women were recommended to visit family doctor and they also got educative booklet with advices for healthy nutrition and physical activity. Implementation of this program indicated the importance of proper lifestyle in the prevention of osteoporosis. Average T-scores of all five groups of women show that osteopenia occurs also in the youngest ones. This indicates the need for a systematic approach to preventing osteoporosis through education of women including younger ones and creating conditions for organized physical activities at the community level.


Retrovirus-mediated WASP gene transfer corrects Wiskott-Aldrich syndrome T-cell dysfunction.
The Wiskott-Aldrich syndrome (WAS) is an X-linked disorder characterized by thrombocytopenia, eczema, and immunodeficiency. At present, the only definitive therapy for the disease is allogeneic bone marrow transplantation (BMT). Because of the frequent lack of suitable donors and the potential severe complications associated with BMT, the development of gene-based therapeutic strategies for WAS is highly desirable. To study whether corrective gene transfer into WAS T cells can lead to restoration of the immunologic defects of WAS, a retroviral vector expressing the WAS protein (WASP) gene was used to transduce human T-lymphotropic virus type 1-transformed T-cell lines and primary T lymphocytes from patients with WAS. After transduction, WAS T cells showed levels of WASP expression similar to those found in cells from normal individuals. In addition, the reconstituted WASP interacted in vitro with proteins containing SH3 domain such as Grb2, PLC-gamma1, and Fyn, each of which are connected to signaling pathways linked to the actin cytoskeleton. Furthermore, after CD3 cross-linking, transduced WAS T lines showed improvement of actin polymerization and T-cell receptor/CD3 down-regulation. More importantly, primary WAS T lymphocytes transduced with WASP acquired the ability to proliferate in response to anti-CD3 stimulation. These findings suggest that biologic defects of WAS T cells can be corrected in vitro by retrovirus-mediated gene transfer and pose the basis for future investigation of gene therapy as treatment for WAS.