PCR detection of amplified 132 bp repeats in Marek's disease virus type 1 (MDV-1) DNA can serve as an indicator for critical genomic rearrangement leading to the attenuation of virus virulence.
A radioactive PCR test was developed that amplified the very virulent Marek's disease virus-1 (vvMDV-1) DNA sequence containing the 132 bp repeats. In apathogenic MDV-1 (CVI 988, Rispens), amplified DNA bands containing multiple copies of 132 bp repeats were identified. In the present study this PCR technique was used to monitor the passage level of vvMDV-1 in chicken embryo fibroblasts (CEF) in which the number of tandem 132 bp repeats was increased. It was found that at passage level 32 of vvMDV-1-B isolate, the 132 bp tandem repeat was already markedly amplified and its pattern resembled that of the MDV-1 (CVI 988, Rispens) vaccine virus DNA. In the vvMDV-1Z strain, amplification of the 132 bp repeat was not detectable at a similar passage level. The PCR test demonstrated that the apathogenic MDV-1 Md11/75c virus developed by extensive in vitro passaging has amplified 132 bp DNA repeats similar to those of the commercial vaccine virus (CVI 988, Rispense). It was also found that the pattern of viral RNA from infected cells detectable by Northern blot hybridization was markedly changed from a 2.4 kb RNA species in cells infected with vvMDV-1 viruses, to four RNA species (ranging from 2.2 to 4.4 kb) in cells infected with passage 32 of MDV-1-B strain, to a very large number of undefined RNA species synthesized in cells infected with attenuated MDV-1 viruses (CVI 988, Rispens and Md 11/75c).


The significance of transient changes in trabecular bone remodeling activation.
After menopause, bone turnover is increased due to an increase in the activation of bone remodeling basic multicellular units (BMUs). The importance of changes in BMU activation to bone histomorphometry data and bone volume has not received adequate attention by many skeletal researchers. Therefore, the influence of BMU activation on the above parameters was modeled by way of computer simulation, and the results were compared to data from published post-menopausal bone loss studies. Using control theory concepts, the increase in BMU activation after menopause was modeled as a zero-, first-, or underdamped (oscillatory) second-order transient BMU activation response to the step input: the decline in estrogen. A computer simulation was developed to model the influence of these three transient BMU activation responses on quantitative histological surface parameters and bone volume. The transient BMU activation responses doubled the number of active BMUs. All three types of transient BMU activation responses produced a rapid 5% decline in bone volume due to increased remodeling space. Oscillations in bone volume and histologic surface parameters over time, similar in nature to those seen in studies of ovariectomized animals, were predicted by the simulation for the oscillatory activation response examined, underdamped second order. An oscillatory BMU activation response may explain some of the transient events during menopause. The increased coherence of BMUs created by such a response may increase the likelihood of trabecular perforations. the inherent nature of an oscillatory activation response may cause its detection to be overlooked and bone remodeling data to be misinterpreted.(ABSTRACT TRUNCATED AT 250 WORDS)


Impact of gastric banding on plasma ghrelin, growth hormone, cortisol, DHEA and DHEA-S levels.
Several endocrine abnormalities are reported in obesity. Some are considered as causative factors, whereas others are considered to be secondary effects of obesity. In the current study, we explored the changes in cortisol, growth hormone (GH), DHEA, DHEA-S and GH releasing hormone (ghrelin) plasma levels in morbidly obese subjects who lost abundant weight following laparoscopic adjustable gastric banding (LAGB). 12 morbidly obese adult patients (15 females), age 21-56 years with BMI 46.0+/-4.4 kg/cm(2), were studied. Blood samples were collected before, 6 and 14 months after LAGB. The levels of DHEA, DHEA-S, cortisol, GH, and ghrelin were determined by commercial kits. Statistical analysis was based on one-way repeated measures ANOVA, followed by Student-Newman-Keuls post-hoc test. Mean BMI reduced significantly along the study course (P=.000). Cortisol plasma levels significantly decreased 6 months after surgery (from 541.4+/-242.4 nM to 382.4+/-142.1 nM, P=.004), but did not change further after 14 months (460.2+/-244.9 nM), despite further reduction in BMI (P=.050). GH constantly increased throughout the study from 0.076+/-0.149 ng/ml, to 0.410+/-0.509 ng/ml at 6 months (NS), to 1.224+/-1.738 ng/ml at 14 months after surgery (P=.001). DHEA, DHEA-S and ghrelin plasma levels remained stable throughout the study. GH levels showed a persistent increase during the 14 months following LAGB in association with the weight loss, while a transient decrease in cortisol levels occurred at the 6-months time-point. In contrast, ghrelin, DHEA and DHEA-S were not altered after surgery. The association between GH and cortisol secretion and surgical- and nonsurgical-induced weight reduction merits further investigation.


Pregnancies and births resulting from in vitro fertilization: French national registry, analysis of data 1986 to 1990. FIVNAT (French In Vitro National).
To describe the characteristics of pregnancies and children at birth resulting from IVF-ET and to assess whether they differ from those after natural conception. Prospective multicenter survey. A national registry, representing 80% of all French IVF-ET activity for the period 1986 to 1990. Data on 7,024 pregnancies, 5,371 deliveries, and 6,879 newborn infants. A form is completed for every clinical pregnancy. Spontaneous abortion, ectopic pregnancy, multiple pregnancy, prematurity, hypotrophy, perinatal and neonatal mortality, and congenital malformations rates. The spontaneous abortion and ectopic pregnancy rates were, respectively, 17.6% and 5.8%. More than a quarter (26.8%) of the deliveries were multiple births. The preterm birth rate (29.3%), the low birth weight rate (36.2%), and the perinatal and neonatal mortality rates were higher than the national average. The rate of malformations (2.8%) was comparable with the general population (2.1%). Assisted conception frequently results in multiple pregnancy, and this is the main determinant of pregnancy outcome. Nevertheless, it is not the only risk factor for prematurity, which is also more frequent among IVF singleton newborn. The prevalence of congenital malformations is not higher than after natural conception.


Development of Bis-cyclic Imidazolidine-4-one Derivatives as Potent Antibacterial Agents.
Antibiotic resistance has emerged as one of the biggest public health concerns all over the world. In an effort to combat bacterial infections, a series of imidazolidine-4-one derivatives with potent and broad-spectrum antibacterial activity and low rates of drug resistance were developed by mimicking the salient physiochemical features of host defense peptides. These small molecules displayed potent activity against both Gram-negative and Gram-positive bacteria including several multidrug-resistant bacteria strains. Meanwhile, time-kill kinetics and drug resistance studies suggested that the most potent compound 3 could not only eliminate the bacteria rapidly but also exhibit a low probability of drug resistance in MRSA over many passages. Further mechanistic studies suggested that 3 eradicated bacterial pathogens by disintegrating membranes of both Gram-negative and Gram-positive bacteria. Together with their small molecular weight and low production cost compared with HDPs, these imidazolidine-4-one compounds may be developed into a new generation of antibiotic therapeutics combating emergent drug resistance.


Generation of induced pluripotent stem cells (iPSCs) from an infant with Pompe disease carrying with compound mutations of R608X and E888X in GAA gene.
Induced pluripotent stem cells (iPSCs) were generated from peripheral blood mononuclear cells (PBMCs) isolated from the peripheral blood of a five months-old boy with glycogen storage disease type II(GSD II, also known as Pompe disease, PD) carries compound mutations R608X E888X in GAA gene. PBMCs were reprogrammed using non-integrative Sendai viral vectors containing reprogramming factors OCT4, SOX2, KLF4 and C-MYC. iPSCs were shown to express pluripotent markers, have trilineage differentiation potential, carry GAA-R608X and GAA-E888X compound mutations, have a normal karyotype. It is useful tool for studying GSDII.


A new species of Bembidion Latrielle 1802 from the Ozarks, with a review of the North American species of subgenus Trichoplataphus Netolitzky 1914 (Coleoptera, Carabidae, Bembidiini).
A new species of Bembidion (Trichoplataphus Netolitzky) from the Ozark Plateau of Missouri and Arkansas is described (Bembidion ozarkense Maddison and Hildebrandt). It is distinguishable from the closely related species, Bembidion rolandi Fall, by characteristics of the male genitalia, and sequences of the genes cytochrome oxidase I and 28S ribosomal DNA. A brief review of the North American species of Trichoplataphus is presented, including a key to species.


Gross hematuria as a manifestation of infectious mononucleosis.
Gross hematuria is an uncommon manifestation of infectious mononucleosis. This report describes a 19-year-old male with infectious mononucleosis who developed gross hematuria. Infectious mononucleosis should be considered in the differential diagnosis of adolescents with hematuria.


Measuring PI3K lipid kinase activity.
Class IA phosphoinositide-3 kinases (PI3Ks) signaling has recently emerged as a key element in cancer development because of its ability to trigger a complex panoply of cellular responses controlling survival and proliferation. Many cancers show inappropriately activated PI3K pathway, and tumors with high PI3K activity are frequently resistant to traditional chemotherapy. Indeed, preclinical studies demonstrated a prominent role for the PI3K pathway in cancer cell survival and growth, thus validating PI3K as a potential drug target in cancer. The emerging interest in inhibiting PI3Ks in cancer have prompted the aggressive development of new selective PI3K pathway inhibitors as cancer therapy, and many of these molecules are currently in early-phase clinical trials. In this chapter, we describe methods to measure the PI3K lipid kinase activity in vitro, which is the standard procedure to test the efficacy of inhibitors.


Structural activity relationship between Salmonella-mutagenicity and nitro-orientation of nitroazaphenanthrenes.
Nitroazaphenanthrenes (NAphs) and their N-oxides (NAphOs) were synthesized as derivatives with nitrogen atoms in the 1, 4, and 9 positions of phenanthrene rings, and as nitrated derivatives substituted at the 1, 2, 3, 4, 5, 6, 7, and 8 positions of phenanthrene rings. To determine the structure activity relationship of these derivatives, all 19 isomers were bioassayed with Salmonella tester strains. NAphs substituted at the 4, 6, 7 and 8 positions were mutagenic for TA98, and 1-, 2-, and 3-N-9-AphOs, 6-N-1-AphO and 6-N-4-AphO were mutagenic for TA98 and TA100 without the S9 mix, while 5-N-1-AphO and 5-N-9-AphO were non- or weakly mutagenic. Nitrated derivatives, 6-N-4-Aph, 6-N-9-Aph, 6-N-1-AphO, and 6-N-4-AphO, were powerful mutagens for TA98 and TA100. Mutagenicity was enhanced by mutant strains producing nitroreductase, such as YG1021 and 1026, and by those producing O-acetyltransferase, such as YG1024 and 1029. Nitro derivatives substituted at positions 4 and 5 in the phenanthrene rings were perpendicular, while those at positions 2, 3, 6 and 7 were coplanar to the phenanthrene rings. NAphs substituted at the 1 and 8 positions were noncoplanar due to steric hindrance of the aromatic proton at the peri position. On the other hand, 1,5- and 1,8-dinitro-4-azaphenanthrenes showed high mutagenicity for strains TA98 and TA100 in the absence of the S9 mix, and were strongly enhanced by nitroreductase and O-acetyltransferase, over-producing mutants. Therefore, it was found that the mutagenic potency of NAphs and NAphOs was closely associated with the chemical properties and orientation of nitro substitution of aromatic rings.


Biochemical and electron microscopic observations of cytotoxic effect of a fraction from hemolytic streptococcus on Ehrlich ascites carcinoma cells.
A protein fraction (60-F), obtained from cell-free extract of living hemolytic streptococcus, Su-strain, by 50-60% saturation with ammonium sulfate, inhibited the de novo synthesis of nucleic acids and proteins in Ehrlich ascites carcinoma (EAC) cells. 60-F released RNA but not DNA from EAC cells. This cytotoxic or cytolytic effect of 60-F was substantiated morphologically by scanning and transmission electron microscopic (SEM and TEM) observations, showing that 60-F induced cellular changes such as a loss of microvilli, bleb formation, cell deformity and partial gap of cell membrane in EAC cells. In addition, it was found that 60-F was more stable in RPMI-1640 medium supplemented with fetal calf serum than in other various media examined and was sensitive to temperature, pH changes and trypsin digestion.


Sudden death due to adrenal neuroblastoma: child abuse mimic?
Neuroblastoma is one of the most common malignancies in children. It is a tumor that can clinically present with varied symptoms that can mimic other entities, including child abuse. It is important that forensic pathologists are aware of the diseases that can masquerade as child abuse. A case of sudden, unexpected death in a 2-year-old due to previously undiagnosed neuroblastoma is presented. At initial presentation of the child in the emergency department, child abuse was considered as a possible explanation for the sudden demise. Death occurred before a full clinical workup could be completed and an autopsy was performed which unveiled the cause of death.


The effect of Foreign Body Aspiration training on the knowledge level of pupils.
Educators dealing with 0-6 years old children must be individuals who are willing to take all kind of measures in order to ensure the safety of children in educational institutions and playgrounds, providing protection from diseases, and who are able to apply first aid measures in case of an accident. In this study, we aimed to determine the level of knowledge of all students continuing their education in the department for child development, regarding Foreign Body Aspiration (FBA) and the effect of FBA training on their knowledge level. This semi-experimental study was carried out on high-school students (n=123) continuing their education in the department for child development in Corum, Turkey. The data was evaluated with appropriate statistical methods, and p<0.05 was determined as statistically significant. Before the training, 80% of the high-school children knew the importance of age factor in cases of FBA, improving to 92% after training. The increase in the number of students who were aware of the importance of age factor was statistically significant (p<0.05). As a result of the training, the visual presentation and training on models related to FBA have led to an increase in the knowledge level of the students.


Global, regional, and national burden and quality of care index (QCI) of lip and oral cavity cancer: a systematic analysis of the Global Burden of Disease Study 1990-2017.
To measure the quality of care for lip and oral cavity cancer worldwide using the data from the Global Burden of Disease (GBD) Study 2017. After devising four main indices of quality of care for lip and oral cavity cancer using GBD 2017 study's measures, including prevalence, incidence, years of life lost, years lived with disability, and disability-adjusted life years, we utilised principal component analysis (PCA) to determine a component that bears the most proportion of info among the others. This component of the PCA was considered as the Quality-of-Care Index (QCI) for lip and oral cavity cancer. The QCI score was then reported in both men and women worldwide and different countries based on the socio-demographic index (SDI) and World Bank classifications. Between 1990 and 2017, care quality continuously increased globally (from 53.7 to 59.6). In 1990, QCI was higher for men (53.5 for men compared with 50.8 for women), and in 2017 QCI increased for both men and women, albeit a slightly higher rise for women (57.2 for men compared with 59.9 for women). During the same period, age-standardised QCI for lip and oral cavity cancer increased in all regions (classified by SDI and World Bank). Globally, the highest QCI scores were observed in the elderly age group, whereas the least were in the adult age group. Five countries with the least amount of QCIs were all African. In contrast, North American countries, West European countries and Australia had the highest indices. The quality of care for lip and oral cavity cancer showed a rise from 1990 to 2017, a promising outcome that supports patient-oriented and preventive treatment policies previously advised in the literature. However, not all countries enjoyed such an increase in the QCI to the same extent. This alarming finding could imply a necessary need for better access to high-quality treatments for lip and oral cavity cancer, especially in central African countries and Afghanistan. More policies with a preventive approach and paying more heed to the early diagnosis, broad insurance coverage, and effective screening programs are recommended worldwide. More focus should also be given to the adulthood age group as they had the least QCI scores globally.


Comparative electron-microscopic studies of benign hepatoma and icterus in patients on oral contraceptives.
The author compared the electron-microscopic picture of 6 benign hepatomas and 3 specimens of icteric liver tissue in female patients on oral contraceptives. The majority of ultrastructural changes seen were identical in the tumor and non-tumor cases. It is noteworthy that there is an accumulation of filamentous substance of unknown origin in the bile canaliculi and in the Disse's spaces associated with a damage to the cell membrane. The electron-microscopic picture of the sinusoidal endothelial cells suggested an increased protein producing activity. The most conspicuous feature of the tumours was formation of continuous capillaries at the site of the sinusoids. The importance of systematic electron-microscopic investigation in studying the effect of oral contraceptives on the liver cells is stressed by the author.


Novel brain-targeted nanomicelles for anti-glioma therapy mediated by the ApoE-enriched protein corona in vivo.
The interactions between nanoparticles (NPs) and plasma proteins form a protein corona around NPs after entering the biological environment, which provides new biological properties to NPs and mediates their interactions with cells and biological barriers. Given the inevitable interactions, we regard nanoparticle‒protein interactions as a tool for designing protein corona-mediated drug delivery systems. Herein, we demonstrate the successful application of protein corona-mediated brain-targeted nanomicelles in the treatment of glioma, loading them with paclitaxel (PTX), and decorating them with amyloid β-protein (Aβ)-CN peptide (PTX/Aβ-CN-PMs). Aβ-CN peptide, like the Aβ1-42 peptide, specifically binds to the lipid-binding domain of apolipoprotein E (ApoE) in vivo to form the ApoE-enriched protein corona surrounding Aβ-CN-PMs (ApoE/PTX/Aβ-CN-PMs). The receptor-binding domain of the ApoE then combines with low-density lipoprotein receptor (LDLr) and LDLr-related protein 1 receptor (LRP1r) expressed in the blood-brain barrier and glioma, effectively mediating brain-targeted delivery. PTX/Aβ-CN-PMs were prepared using a film hydration method with sonication, which was simple and feasible. The specific formation of the ApoE-enriched protein corona around nanoparticles was characterized by Western blotting analysis and LC-MS/MS. The in vitro physicochemical properties and in vivo anti-glioma effects of PTX/Aβ-CN-PMs were also well studied. The average size and zeta potential of PTX/Aβ-CN-PMs and ApoE/PTX/Aβ-CN-PMs were 103.1 nm, 172.3 nm, 7.23 mV, and 0.715 mV, respectively. PTX was efficiently loaded into PTX/Aβ-CN-PMs, and the PTX release from rhApoE/PTX/Aβ-CN-PMs exhibited a sustained-release pattern in vitro. The formation of the ApoE-enriched protein corona significantly improved the cellular uptake of Aβ-CN-PMs on C6 cells and human umbilical vein endothelial cells (HUVECs) and enhanced permeability to the blood-brain tumor barrier in vitro. Meanwhile, PTX/Aβ-CN-PMs with ApoE-enriched protein corona had a greater ability to inhibit cell proliferation and induce cell apoptosis than taxol. Importantly, PTX/Aβ-CN-PMs exhibited better anti-glioma effects and tissue distribution profile with rapid accumulation in glioma tissues in vivo and prolonged median survival of glioma-bearing mice compared to those associated with PMs without the ApoE protein corona. The designed PTX/Aβ-CN-PMs exhibited significantly enhanced anti-glioma efficacy. Importantly, this study provided a strategy for the rational design of a protein corona-based brain-targeted drug delivery system. More crucially, we utilized the unfavorable side of the protein corona and converted it into an advantage to achieve brain-targeted drug delivery.


Streptococcal intertrigo of the cervical folds in a five-month-old infant.
We present a case of severe intertrigo in the neck of a 5-month-old infant. The cause was Streptococcus pyogenes.


Bacopa monnieri ameliorates memory deficits in olfactory bulbectomized mice: possible involvement of glutamatergic and cholinergic systems.
This study investigated the effects of alcoholic extract of Bacopa monnieri (L.) Wettst. (BM) on cognitive deficits using olfactory bulbectomized (OBX) mice and the underlying molecular mechanisms of its action. OBX mice were treated daily with BM (50 mg/kg, p.o.) or a reference drug, tacrine (2.5 mg/kg, i.p.), 1 week before and continuously 3 days after OBX. Cognitive performance of the animals was analyzed by the novel object recognition test, modified Y maze test, and fear conditioning test. Brain tissues of OBX animals were used for neurochemical and immunohistochemical studies. OBX impaired non-spatial short-term memory, spatial working memory, and long-term fair memory. BM administration ameliorated these memory disturbances. The effect of BM on short-term memory deficits was abolished by a muscarinic receptor antagonist, scopolamine. OBX downregulated phosphorylation of synaptic plasticity-related signaling proteins: NR1 subunit of N-methyl-D-aspartate receptor, glutamate receptor 1 (GluR1), and calmodulin-dependent kinase II but not cyclic AMP-responsive element binding protein (CREB), and reduced brain-derived neurotrophic factor (BDNF) mRNA in the hippocampus. OBX also reduced choline acetyltransferase in the hippocampus and cholinergic neurons in the medial septum, and enlarged the size of lateral ventricle. BM administration reversed these OBX-induced neurochemical and histological alterations, except the decrease of GluR1 phosphorylation, and enhanced CREB phosphorylation. Moreover, BM treatment inhibited ex vivo activity of acetylcholinesterase in the brain. These results indicate that BM treatment ameliorates OBX-induced cognition dysfunction via a mechanism involving enhancement of synaptic plasticity-related signaling and BDNF transcription and protection of cholinergic systems from OBX-induced neuronal damage.


Immunomodulation of in vitro antigen presentation by cations.
The ability of various cations to alter an antigen-specific (ovalbumin) T cell activation system in vitro has been assessed. This in vitro system includes analysis of antigen capture, processing, and presentation by antigen presenting cells (APC) (splenocytes, B cells or macrophages) in a major histocompatibility complex (MHC) restricted fashion and the evaluation of the activation of a T-cell hybridoma, DO-11.10, which is specific for "processed" ovalbumin in association with I-Ad. Activation is determined by production of IL2 which is quantitated in a bioassay with HT-2 cells. Numerous metals (10 and 100 microM) were screened in the coculture assay (APC and DO-11.10 plus ovalbumin). Metals with inhibitory effects were Cd greater than Cu greater than Pb greater than Zn. Co and Cr had no modulatory effect and Ni had an enhancing effect (increased IL2 production). The effects of the modulatory metals were further assessed for influences on the individual cellular components of this system. Cd was toxic to all cell types whereas Cu was toxic only to irradiated splenocytes and Do-11.10. Pb was the only metal which was not toxic to any cell type but still inhibited antigen presentation. To prevent influences of metals on DO-11.10 or carryover into the bioassay, APC were preincubated with metals and then assessed for presentation capability. After preincubation, only Cd, Pb, and Cu were inhibitory. The inhibition by Cd was due to toxicity. Cu inhibited only irradiated splenocyte presentation and this effect was due to toxicity. Pb inhibition was due to a mechanism other than toxicity and its biochemical influences on APC are discussed.


Recent advances in tailoring and improving the properties of polyimide aerogels and their application.
With the rapid advancements in technology and growing aerospace applications, there is a need for effective low-weight and thermally insulating materials. Aerogels are known for their ultra-lightweight and they are highly porous materials with nanopores in a range of 2 to 50 nm with very low thermal conductivity values. However, due to hygroscopic nature and brittleness, aerogels are not used commercially and in daily life. To enhance the mechanical and hydrophobic properties, reinforcement materials such as styrene, cyanoacrylates, epoxy along with hydroxyl, amines, vinyl groups are added to the surface. The addition of organic materials resulted in lower service temperatures which reduce its potential applications. Polyimides (PI) are commonly used in engine applications due to their suitable stability at high temperatures along with excellent mechanical properties. Previous research on polyimide aerogels reported high flexibility or even foldability. However, those works' strategy was mainly limited to altering the backbone chemistry of polyimide aerogels by changing either the monomer's compositions or the chemical crosslinker. This work aims to summarize, categorize, and highlight the recent techniques for improving and tailoring properties of polyimide aerogels followed by the recent advancements in their applications.


Diastereoselective two-directional synthesis and cation transport ability of the central tristetrahydrofuranyl unit of meso polyether glabrescol as naturally occurring podand.
The diastereoselective synthesis of the central 2,5-linked tristetrahydrofuran (trisTHF) 6 of naturally occurring meso polyether, glabrescol (5), has been achieved in a two-directional manner by the vanadium(V)-catalyzed anti oxidative cyclizations of diol 14. The trisTHF podand 6 and its stereoisomeric analogs 7 and 8 exhibited outstanding cation transport abilities for physiologically important Na+ and K+.


Supraorbital Keyhole Approach to the Sella and Anterior Skull Base via a Forehead Wrinkle Incision.
To evaluate safety and effectiveness of supraorbital keyhole approach to the sella and anterior skull base via a forehead wrinkle incision. We analyzed and reclassified forehead wrinkle types in 100 recruited healthy individuals. Twenty patients with different intracranial lesions in the sella and anterior skull base areas were selected for surgery using the supraorbital keyhole approach via a forehead wrinkle incision by forehead wrinkle type. All clinical patient records were retrospectively reviewed. Based on the feasibility of the surgical approach, the forehead wrinkles in Chinese individuals were categorized into 2 types: horizontal linear and nonlinear. Gross total removal of the lesions was achieved in 90% of the cases using this approach. All patients had excellent cosmetic results. Except for 2 patients with poor prognoses, a high level of comfort and satisfaction with the forehead wrinkle incisions was obtained based on the scale scores of pain from scars and headaches and satisfaction with the cosmetic results. No permanent approach-related complications occurred. The supraorbital keyhole approach via a forehead wrinkle incision was safe and effective. This approach provided a sufficient visual field for lesion resection and satisfactory cosmetic results for patients.


Simplified technique for minimally invasive repair of congenital diaphragmatic hernia using hollow-needle snare and transthoracic traction stitches.
Minimally invasive (MI) congenital diaphragmatic hernia (CDH) repair can be challenging. Placing rib-anchoring stitches without creating skin incisions and closing wider defects are some of the difficulties. In Bochdaleck hernia repair, maintaining visceral reduction and minimizing pneumothorax use are additional obstacles. We describe the use of hollow-needle snares (HNS) and transthoracic traction stitches (TTS) to overcome these challenges. Hollow-needle snares is assembled by passing a prolene stitch through a hollow needle creating a retractable snare, which is used to extract the placed anchoring stitches by passing it over the ribs but through the same stitch's skin entrance site. In Bochdaleck hernia, the early placement of TTS using HNS can facilitate visceral reduction, patch lay down, and tension-free closure of possible residual V-shaped defects. Between July 2009 and April 2011, we performed 10 consecutive MI CDH repairs for 9 patients, including 7 Bochdaleck and 3 Morgagni hernias. The median age was 8 days (range, 3-172 days), and the mean operative time was 148.5 ± 37.8 minutes for Bochdaleck hernia repairs. For Morgagni hernia, the median age was 18.3 months (range, 10.5-37 months), and the mean operative time was 100 ± 26.5 minutes. All cases were completed without conversion. One patient had a hernia recurrence and was repaired similarly, whereas the others had uneventful recovery at a median follow-up of 5.8 months (range, 1.1-23.7 months). Hollow-needle snare and TTS are simple and available tools that can facilitate MI repair of CDH. This initial experience demonstrates the technique's effectiveness and its excellent cosmetic outcomes.


Phylogenetic placement of the enigmatic and critically endangered genus Saniculiphyllum (Saxifragaceae) inferred from combined analysis of plastid and nuclear DNA sequences.
Saniculiphyllum, a monotypic genus distributed in Southwest China, was thought to be extinct before our recent rediscovery. The taxonomic position of this genus has been enigmatic ever since its publication. It was originally treated as the only member of a distinct tribe Saniculiphylleae in the family Saxifragaceae. Some proposed a new family, Saniculophyllaceae, to accommodate this genus, although its affinities are clearly with members of Saxifragaceae. Here we analyzed six DNA regions, the nuclear ribosomal ITS and 26S rDNA and the plastid rbcL, matK, trnL-trnF, psbA-trnH genes, spacers, and intron to explore the phylogenetic position of Saniculiphyllum within Saxifragaceae. The combined nuclear and chloroplast dataset includes 63 ingroup species, representing all genera but Hieronymusia in the family. Results from likelihood, parsimony and Bayesian phylogenetic methods corroborate earlier results. Two clades of Saxifragaceae, the Heucheroid and Saxifragoid clades, were recovered. The topologies obtained from different analyses confirm the placement of Saniculiphyllum in Saxifragaceae, but our analyses reveal that Saniculiphyllum is embedded within the large Heucheroid clade. However, the closest relatives of Saniculiphyllum within the Heucheroid clade remain unclear. Combined with morphological data, our results suggest that Saniculiphyllum should best be regarded as a highly distinctive lineage within the Heucheroid clade of Saxifragaceae. Morphological novelties and conservation status of Saniculiphyllum are also presented.


Τhe genetics of juvenile idiopathic arthritis: Searching for new susceptibility loci.
Juvenile idiopathic arthritis (JIA) is an autoimmune disease that is characterized by persistent chronic arthritis and affected by genetic and environmental factors. Different genetic variations have been reported as risk factors for JIA. However, given that many results could not be replicated in individuals of different ancestral origin, it was assumed that heterogeneous genetic factors are involved in this disease. In the present study, we analyzed three single nucleotide polymorphisms (SNPs), namely PTPRC (rs10919563), TYK2 (rs34536443) and PRKCQ (rs4750316), which were found to be associated with JIA in previous studies. We also investigated whether the intron‑4 located 27‑bp VNTR of endothelial nitric oxide synthase (eNOS), is associated with risk for JIA in Greece. In total, 125 JIA patients and 221 healthy controls from northern Greece were included in the study as a sample set. Samples were then analyzed, and genotyped for the three SNPs with TaqMan primer‑probe sets, using a Real‑Time PCR platform (ViiA™ 7 Real‑Time PCR system), while eNOS VNTR polymorphism was genotyped by PCR. Statistical analysis was performed using a GraphPad Prism statistical program. The χ2 test was used to examine differences of genotype and allele frequencies between patients and controls. Statistical significance was defined by using the two‑tailed P<0.05 test. Bioinformatics analysis was conducted by using BlastP, Pymol, Maestro and Desmond. In the case‑control association study performed, eNOS only was found to be associated with JIA. Genotype a/a and allele 'a' were found in a higher frequency in JIA patients than in controls [p<0.0001, odds ratio (OR)=0.15, 95% confidence intervals (CI): 0.065‑0.37; and p<0.0001, OR=0.34, 95% CI: 0.23‑0.49, respectively]. No associations with JIA were detected for TYK2, PTPRC or PRKCQ. Aiming to investigate the structural consequences and the structure/function relationships accompanying the Pro1104 to Ala (rs34536443) mutation on TYK2 protein, bioinformatics analysis was performed. Combining three‑dimensional (3D)‑modeling and molecular dynamics simulations we identified changes in structural flexibility, affecting the functionality of the kinase domain of TYK2. To the best of our knowledge, this is the first time that eNOS VNTR polymorphism is associated with susceptibility to JIA, suggesting a differential role of allele 'a' in various complex diseases. The current data emphasize the importance of comparative studies in populations of a different ancestral background towards the clarification of the role of specific alleles in the development of JIA.


Moving fluid with bacterial carpets.
We activated a solid-fluid interface by attaching flagellated bacteria to a solid surface. We adsorbed swarmer cells of Serratia marcescens to polydimethylsiloxane or polystyrene. The cell bodies formed a densely packed monolayer while their flagella continued to rotate freely. Motion of the fluid close to an extended flat surface, visualized with tracer beads, was dramatically enhanced compared to the motion farther away. The tracer beads revealed complex ever-changing flow patterns, some linear (rivers), others rotational (whirlpools). Typical features of this flow were small (tens of micro m) and reasonably stable (many minutes). The surface performed active mixing equivalent to diffusion with a coefficient of 2 x 10(-7) cm(2)/s. We call these flat constructs "bacterial carpets". When attached to polystyrene beads or to fragments of polydimethylsiloxane, the bacteria generated both translation and rotation. We call these constructs "auto-mobile beads" or "auto-mobile chips". Given the size and strength of the flow patterns near the carpets, the motion must be generated by small numbers of coordinated flagella. We should be able to produce larger and longer-range effects by increasing coordination.


Effectiveness of antibiotic-loaded bone cement in total joint arthroplasty at a tertiary medical center: A retrospective cohort study.
Data regarding the effectiveness of antibiotic-loaded bone cement (ALBC) in preventing prosthetic joint infections (PJI) after total joint arthroplasty (TJA) is inconsistent. The objective of this study was to evaluate if the routine use of ALBC influenced the risk of revision surgery due to PJI. This is a retrospective cohort study performed between January 2018 and September 2020. Adult patients aged ≥ 18 years who underwent TJA (knee or hip) and received either ALBC or plain cement (PC) were included. The outcome of this study was the rate of revision due to PJI. Multivariate analysis using logistic regression was used to identify factors that may be associated with increased risk of PJI, using STATA 15.1 (StataCorp LP, College Station, Texas, USA). A total of 844 patients were screened and 319 patients were included. There were 247 patients in ALBC group and 72 patients in the PC group. Only vancomycin powder was used in all ALBC cases, with a 2 g dose in 50% of the cases (dose ranged between 1 g and 8 g). The status of the prosthetic joint was assessed and recorded up to 2 years of the TJA. Overall, the difference in the rates of PJI between the two groups after primary arthroplasty was not statistically significant (5.6% vs 1.4%; p = 0.173; OR, 4.2; 95% CI, 0.5-33). ALBC was not associated with a reduction in PJI rates after primary TJA. More research is needed to further evaluate the effectiveness of ALBC in preventing PJI.


Protein quality in cereals and pulses. 3. Bioassays with rats and chickens on sorghum (Sorghum vulgare Pers.), barley and field beans (Vicia faba L.). Influence of polyethylene glycol on digestibility on the protein in high-tannin grain.
1. Two preceding papers in this series describe the application of microbiological and other in vitro tests in the evaluation of sorghum (Sorghum vulgare Pers.), field beans (Vicia faba L.) and barley, and in assessing the influence of polyethylene glycol (PEG 4000) on the nutritional availability of the methionine. The present paper gives for comparison the results of bioassays on some of the same test samples. Net protein utilization (NPU) in rats was measured by the nitrogen balance method, and N digestibility in chickens by the ileal analysis procedure. 2. In rat tests on sorghum, N in grain of high-tannin varieties was poorly digested. Supplementation of the test diets with 0.1 g PEG 4000/g protein gave a large improvemnet, which was partly offset by an apparent decrease in biological value (BV). With chickens N digestibility was even lower, and was similarly improved with PEG 4000. Treatment of high-tannin grain with ammonia solution was also effective in improving N digestibility. 3. With low-tannin sorghum the amino acid digestibilities were uniformly high and were not affected by addition of PEG to the test diet. With high-tannin sorghums they were low and less uniform, and were much improved by PEG 4000. 4. With field beans, the influence of the seed-coat tannin on protein utilization was much less pronounced than with sorghum. In chickens there was a significant effect (P less than 0.05) of PEG 4000 on N idgestibility in a high-tannin variety. With rats the effect was smaller and not significant. 5. In four samples of barley. N digestibility was high (0.87--0.96) and was not further improved by PEG 4000. The BV of a high-lysine cultivar proved marginally inferior to that of a normal variety. Possible reasons for this are discussed. 6. Over all, the results were closely consistent with those from microbiological tests with Streptococcus zymogenes.


Combined clarification and affinity capture using magnetic resin enables efficient separation of rAAV5 from cell lysate.
Recombinant adeno-associated virus (rAAV) vectors have displayed enormous potential as a platform for delivery of gene therapies. Purification of rAAV at industrial scale involves a series of elaborate, material, and time-consuming midstream steps, such as clarification by depth filtration and concentration/buffer exchange by tangential flow filtration. In this study, we developed a filter-less flow capture method for purification of rAAV serotype 5, using a high-gradient magnetic separator and magnetic Mag Sepharose beads coupled to an AVB affinity ligand. In under 2 h, we captured and eluted rAAV5 directly from ∼5 L of cell lysate with a recovery yield of 63% (±5%, n = 3). Compared to cell lysate, the eluate showed a 3-log reduction of host cell DNA and host cell proteins. The process developed eliminates the need for filtration and column chromatography in the early steps of industrial rAAV purification. This will be of high value for industrial-scale manufacturing of rAAVs by reducing time and material in the purification process, without compromising product recovery and purity.


Atherogenic LOX-1 signaling is controlled by SPPL2-mediated intramembrane proteolysis.
The lectin-like oxidized LDL receptor 1 (LOX-1) is a key player in the development of atherosclerosis. LOX-1 promotes endothelial activation and dysfunction by mediating uptake of oxidized LDL and inducing pro-atherogenic signaling. However, little is known about modulators of LOX-1-mediated responses. Here, we show that the function of LOX-1 is controlled proteolytically. Ectodomain shedding by the metalloprotease ADAM10 and lysosomal degradation generate membrane-bound N-terminal fragments (NTFs), which we identified as novel substrates of the intramembrane proteases signal peptide peptidase-like 2a and b (SPPL2a/b). SPPL2a/b control cellular LOX-1 NTF levels which, following self-association via their transmembrane domain, can activate MAP kinases in a ligand-independent manner. This leads to an up-regulation of several pro-atherogenic and pro-fibrotic targets including ICAM-1 and the connective tissue growth factor CTGF. Consequently, SPPL2a/b-deficient mice, which accumulate LOX-1 NTFs, develop larger and more advanced atherosclerotic plaques than controls. This identifies intramembrane proteolysis by SPPL2a/b as a novel atheroprotective mechanism via negative regulation of LOX-1 signaling.


A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA).
The expression of the genes for the major outer membrane proteins OmpF and OmpC are osmoregulated. The ompC locus was found to be transcribed bidirectionally under conditions of high osmolarity and a 174-base transcript encoded upstream of ompC was found to inhibit the OmpF production and to substantially reduce the amount of the ompF mRNA. This RNA [mRNA-interfering complementary RNA (micRNA)] has a long sequence that is complementary to the 5' end region of the ompF mRNA. We propose that the micRNA inhibits the translation of the ompF mRNA by hybridizing with it. This RNA interaction may cause premature termination of the transcription of the ompF gene or destabilization of the ompF mRNA or both.


Patch detachment after mitral valve repair with posterior leaflet augmentation: a case report.
Mitral valve (MV) repair is indicated for patients with severe MR. We report a case of acute MR caused by patch detachment after posterior leaflet augmentation in MV repair. A 65-year-old male underwent MV repair with posterior leaflet augmentation and coronary artery bypass graft 1 month prior to this study. An inverted T-shaped incision was made on the posterior mitral leaflet (PML), and a piece of autologous fresh pericardium was sewn in the PML defect. Seven days after hospital discharge, he started feeling chest pain and presented with pulseless electrical activity. Ultrasonic cardiography showed severe mitral regurgitation (MR), which was suggestive of acute MR. We performed emergency reoperation. The edge of the autologous pericardial patch was detached from the anterior papillary muscle, and MV replacement was performed. He was discharged from the hospital 55 days after the reoperation and returned to his normal daily life. We conclude that avoidance of tension focalization during MV repair may be important.


Methane Carboxylation Using Electrochemically Activated Carbon Dioxide.
Direct synthesis of CH3 COOH from CH4 and CO2 is an appealing approach for the utilization of two potent greenhouse gases that are notoriously difficult to activate. In this Communication, we report an integrated route to enable this reaction. Recognizing the thermodynamic stability of CO2 , our strategy sought to first activate CO2 to produce CO (through electrochemical CO2 reduction) and O2 (through water oxidation), followed by oxidative CH4 carbonylation catalyzed by Rh single atom catalysts supported on zeolite. The net result was CH4 carboxylation with 100 % atom economy. CH3 COOH was obtained at a high selectivity (>80 %) and good yield (ca. 3.2 mmol g-1 cat in 3 h). Isotope labelling experiments confirmed that CH3 COOH is produced through the coupling of CH4 and CO2 . This work represents the first successful integration of CO/O2 production with oxidative carbonylation reaction. The result is expected to inspire more carboxylation reactions utilizing preactivated CO2 that take advantage of both products from the reduction and oxidation processes, thus achieving high atom efficiency in the synthesis.


Age-related pulmonary crackles (rales) in asymptomatic cardiovascular patients.
The presence of age-related pulmonary crackles (rales) might interfere with a physician's clinical management of patients with suspected heart failure. We examined the characteristics of pulmonary crackles among patients with stage A cardiovascular disease (American College of Cardiology/American Heart Association heart failure staging criteria), stratiffed by decade, because little is known about these issues in such patients at high risk for congestive heart failure who have no structural heart disease or acute heart failure symptoms. After exclusion of comorbid pulmonary and other critical diseases, 274 participants, in whom the heart was structurally (based on Doppler echocardiography) and functionally (B-type natriuretic peptide <80 pg/mL) normal and the lung (X-ray evaluation) was normal, were eligible for the analysis. There was a significant difference in the prevalence of crackles among patients in the low (45-64 years; n = 97; 11%; 95% CI, 5%-18%), medium (65-79 years; n = 121; 34%; 95% CI, 27%-40%), and high (80-95 years; n = 56; 70%; 95% CI, 58%-82%) age-groups (P <.001). The risk for audible crackles increased approximately threefold every 10 years after 45 years of age. During a mean follow-up of 11 +/- 2.3 months (n = 255), the short-term (< or =3 months) reproducibility of crackles was 87%. The occurrence of cardiopulmonary disease during follow-up included cardiovascular disease in 5 patients and pulmonary disease in 6. Recognition of age-related pulmonary crackles (rales) is important because such clinically unimportant crackles are so common among elderly patients that, without knowledge of this phenomenon, their existence might interfere with the physician's management of cardiopulmonary patients.


Antibodies against basic fibroblast growth factor inhibit the autocrine growth of pulmonary artery endothelial cells.
Anti-recombinant human basic fibroblast growth factor (rbFGF) antibodies efficiently inhibited the basal proliferation of bovine pulmonary artery endothelial (BAE) cells. The cell-free extract of BAE cells stimulated the proliferation of bovine capillary endothelial cells and this activity was completely abolished by the antibodies. Furthermore, on heparin HPLC the activity was eluted at exactly the same retention time as that for authentic pituitary bFGF. These observations directly indicate that the BAE cells produce bFGF that stimulates their own basal growth by binding to specific receptors expressed on the cell surface.


Medical comorbidities predict the need for colectomy for complicated and recurrent diverticulitis.
The objective of this study was to identify risk factors for recurrent or complicated diverticulitis requiring colectomy. A total of 112 patients were admitted to the West Haven Veterans Affairs Medical Center with the diagnosis of colonic diverticular disease from January 1998 to December 2006. Patients' records were assessed for demographics, past medical history, and physical and biochemical features of presentation. Student t tests, analysis of variance, and chi-square analysis were used to compare binary and categoric data. The medical records of 112 patients admitted to the West Haven Veterans Affairs hospital with the diagnosis of diverticulitis were analyzed retrospectively. A total of 97.3% were male (n = 109), with a mean age of 63.3 years, and a significant smoking history greater than 30 pack-years was present in 70.5% (n = 79) of patients. Eighty-four percent (n = 94) of patients presented with localized abdominal pain and 69.6% (n = 78) had abdominal tenderness without peritoneal signs. Computed tomography was performed in 85.7% (n = 96) of cases. A total of 23.2% (n = 26) of patients proceeded to laparotomy, with free perforation being the most common indication (38.4%, n = 12) followed by a history of 2 or more antecedent attacks of diverticulitis (23.1%, n = 6). Analysis of variance showed that serum albumin levels were significantly lower in the group undergoing colectomy compared with those who did not (3.4 vs 4.1 mg/dL; P = .016). The need for colectomy owing to complicated or recurrent attacks correlated with glucocorticoid use (P < .001) and a history of chronic obstructive pulmonary disease (P < .038), but not with diabetes mellitus, collagen vascular disease, or inflammatory bowel disease. The rules regarding the treatment of diverticulitis are evolving. Comorbid conditions such as hypoalbuminemia, chronic obstructive pulmonary disease, and glucocorticoid use may predispose patients to recurrent or complicated attacks of diverticulitis requiring colectomy. Stratification and reduction of risks may reduce the overall morbidity and mortality of diverticulitis.


Omega-3 fatty acids: how can they be used in secondary prevention?
Omega-3 fatty acids (FAs) are divided into long-chain fatty acids (eicosapentaenoic acid and docosahexaenoic acid ), which are found in fatty fish, and intermediate-chain FAs (alpha-linolenic acid), which are found in vegetable oils. Omega-3 FAs favorably modulate a variety of vascular risk factors and also exert antiarrhythmic effects. Epidemiologic data suggest that increased consumption of marine omega-3 FAs is associated with reduced coronary heart disease (CHD) mortality. Randomized controlled studies also show that supplementation with EPA and DHA reduces CHD risk, primarily in the secondary prevention setting. Data are more limited on the efficacy of marine omega-3 FAs for the primary prevention of CHD and on the role of alpha-linolenic acid. Increased intake of EPA and DHA represents a valuable tool for vascular disease prevention and should be recommended in all patients with CHD.


[3H]-acetylcholine release from rat atria in chronic chagasic cardiopathy.
Isolated superfused rat atria release [3H]-acetylcholine when depolarized with 57 mM potassium. The depolarization-induced [3H]-acetylcholine overflow was significantly reduced in atria from chronically T. cruzi-infected rats with electrocardiographically characterized cardiopathy. This fact suggests the occurrence of functional alterations of cardiac parasympathetic control in these animals, probably related to cardiac ganglion cell destruction.


Simvastatin exerts antifibrotic effect and potentiates the antischistosomal effects of praziquantel in a murine model: Role of IL10.
Previous studies on simvastatin use in experimental schistosomiasis in mice did not provide a full explanation of its mechanism as antischisome. In this study, we tried to find out the role of IL-10 in the mechanism of action of simvastatin. We used 50 clear mice. Ten were used as normal not treated while 40 were infected with shistosome mansoni then divided into 4 groups; 3 treated groups by praziquantel, simvastatin and combined (praziquantel plus simvastatin) respectively and one group non-treated. The simvastatin treated group showed shortening and loss of the tubercles and disappearance of the spines with swelling and twisted shape of the worms. In addition, it also showed mitigation of ovideposition activity of the worms in the liver and reduction of the fibrous component of the liver granuloma producing a protective effect on the liver. This effect was associated with lowering of IL-10. This may explain the role of IL-10 in the protective effect of simvastatin. Combination of treatment with simvastatin plus praziquantel produced more significant effects in different parameters compared with praziquantel treated group. We recommend using simvastatin as add on therapy to standard antischistosomal therapy, praziquantel. Both drugs affect the worm motility and sucker activity and the ova deposition. Simvastatin has an additional pleiotropic effect halting inflammation and decreasing fibrosis due to increasing IL-10 leading to a hepatoprotective effect. Further clinical studies are needed to further validate these findings.


Genome sequence analysis of potential probiotic strain Leuconostoc lactis EFEL005 isolated from kimchi.
Leuconostoc lactis EFEL005 (KACC 91922) isolated from kimchi showed promising probiotic attributes; resistance against acid and bile salts, absence of transferable genes for antibiotic resistance, broad utilization of prebiotics, and no hemolytic activity. To expand our understanding of the species, we generated a draft genome sequence of the strain and analyzed its genomic features related to the aforementioned probiotic properties. Genome assembly resulted in 35 contigs, and the draft genome has 1,688,202 base pairs (bp) with a G+C content of 43.43%, containing 1,644 protein-coding genes and 50 RNA genes. The average nucleotide identity analysis showed high homology (≥ 96%) to the type strain L. lactis KCTC3528, but low homology (≤ 95%) to L. lactis KCTC3773 (formerly L. argentinum). Genomic analysis revealed the presence of various genes for sucrose metabolism (glucansucrases, invertases, sucrose phosphorylases, and mannitol dehydrogenase), acid tolerance (F1F0 ATPases, cation transport ATPase, branched-chain amino acid permease, and lysine decarboxylase), vancomycin response regulator, and antibacterial peptide (Lactacin F). No gene for production of biogenic amines (histamine and tyramine) was found. This report will facilitate the understanding of probiotic properties of this strain as a starter for fermented foods.


Resistance properties and the role of the inner membrane and coat of Bacillus subtilis spores with extreme wet heat resistance.
A protein termed 2Duf greatly increases wet heat resistance of Bacillus subtilis spores. The current work examines the effects of 2Duf on spore resistance to other sporicides, including chemicals that act on or must cross spores' inner membrane (IM), where 2Duf is likely present. The overall aim was to gain a deeper understanding of how 2Duf affects spore resistance, and of spore resistance itself. 2Duf's presence increased spore resistance to chemicals that damage or must cross the IM to kill spores. Spore coat removal decreased 2Duf-spore resistance to chemicals and wet heat, and 2Duf-spores made at higher temperatures were more resistant to wet heat and chemicals. 2Duf-less spores lacking coats and Ca-dipicolinic acid were also extremely sensitive to wet heat and chemicals that transit the IM to kill spores. The new work plus previous results lead to a number of important conclusions as follows. (1) 2Duf may influence spore resistance by decreasing the permeability of and lipid mobility in spores' IM. (2) Since wet heat-killed spores that germinate do not accumulate ATP, wet heat may inactivate some spore IM protein essential in ATP production which is stabilized in a more rigid IM. (3) Both Ca-dipicolinic acid and the spore coat play an important role in the permeability of the spore IM, and thus in many spore resistance properties. The work in this manuscript gives a new insight into mechanisms of spore resistance to chemicals and wet heat, to the understanding of spore wet heat killing, and the role of Ca-dipicolinic acid and the coat in spore resistance.


Blood glucose and plasma insulin responses to fat-free milk and low-lactose fat-free milk in young type 1 diabetics.
The blood glucose and plasma insulin responses to test milk samples were studied in eight insulin-dependent diabetics. After an overnight fast, the subjects (aged 20-45 years) were given a breakfast containing two Weetabix biscuits (20 g carbohydrate) with 500 ml of either regular (S) fat-free milk (25 g lactose) or 500 ml of a new low-lactose (D) fat-free milk (3.75 g lactose and 4.25 g fructose). The regular morning insulin dose was omitted. Mean basal plasma glucose level was slightly higher before S milk (11.4 vs. 10.1 mmol/l). The peak increment in plasma glucose was higher in S milk (9.4 vs. 6.6 mmol/l). The rise was 83% above basal (S) vs. 65% (D). Although the final mean plasma glucose concentration was not significantly higher 3 h after S milk compared with D milk (17.9 vs. 14.3 mmol/l) the incremental area under the plasma glucose curve was much greater (p less than 0.001) with S milk than with D milk (1266 +/- 295 units vs. 909 +/- 242 units). No galactose was detectable in any sample and there was no difference in plasma beta-hydroxybutyrate levels. The results suggest that the low-lactose fat-free milk (D) may be suitable for diabetic diets and weight reducing diets due to the lower contribution of energy. The results suggest that fat-free milk does not exert a fast action on blood glucose concentration and therefore fat-free milk and especially low-lactose fat-free milk may also prove to be suitable for diabetic diets.


Efficacy and safety of interferon beta-1b sc in older RRMS patients--a posthoc analysis of the BEYOND study.
Evidence of a significant improvement of IFNB-1b in clinical severity in the older population with RRMS has not been established so far. The aim of this exploratory post hoc analysis of the 250 mcg IFNB-1b group of the BEYOND study is to compare the efficacy and safety of older versus younger patients using a cut-off at the age of 50 and at the age of 40, respectively. There was no difference between age groups in adjusted relapse risk (age 50 cut-off: P = 0.482, age 40 cut-off: P = 0.073) nor in adjusted time to confirmed EDSS progression (age 50 cut-off: P = 0.096, age 40 cut-off: P = 0.189). There were no significant differences between patients <50 and ≥ 50 years in the adjusted annualized relapse rate (P = 0.285), whereas relapse rate was higher in the <40 as compared to the ≥ 40 group (P = 0.024). The proportion of patients with confirmed disability progression was not significantly different for the 50 cutoff (P = 0.148), whereas significantly fewer <40 than ≥ 40 patients had disability progression (P = 0.047). Only minor differences in adverse event frequencies between the age groups for the two cut-offs were seen. These results indicate that IFNB-1b is as efficacious and safe in patients ≥ 50 years as <50 years of age.


Occlusion of activity dependent synaptic plasticity by late hypoxic long term potentiation after neonatal intermittent hypoxia.
To elucidate the mechanisms of memory impairment after chronic neonatal intermittent hypoxia (IH), we employed a mice model of severe IH administered at postnatal days 3 to 7. Since prior studies in this model did not demonstrate increased cell death, our primary hypothesis was that IH causes a functional disruption of synaptic plasticity in hippocampal neurons. In vivo recordings of Schaffer collateral stimulation-induced synaptic responses during and after IH in the CA1 region of the hippocampus revealed pathological late phase hypoxic long term potentiation (hLTP) (154%) that lasted more than four hours and could be reversed by depotentiation with low frequency stimulation (LFS), or abolished by NMDA and PKA inhibitors (MK-801 and CMIQ). Furthermore, late phase hLTP partially occluded normal physiological LTP (pLTP) four hours after IH. Early and late hLTP phases were induced by neuronal depolarization and Ca2+ influx, determined with manganese enhanced fMRI, and had increased both AMPA and NMDA - mediated currents. This was consistent with mechanisms of pLTP in neonates and also consistent with mechanisms of ischemic LTP described in vitro with OGD in adults. A decrease of pLTP was also recorded on hippocampal slices obtained 2 days after IH. This decrease was ameliorated by MK-801 injections prior to each IH session and restored by LFS depotentiation. Occlusion of pLTP and the observed decreased proportion of NMDA-only silent synapses after neonatal hLTP may explain long term memory, behavioral deficits and abnormal synaptogenesis and pruning following neonatal IH.


Human effector and memory CD8+ T cell responses to smallpox and yellow fever vaccines.
To explore the human T cell response to acute viral infection, we performed a longitudinal analysis of CD8(+) T cells responding to the live yellow fever virus and smallpox vaccines--two highly successful human vaccines. Our results show that both vaccines generated a brisk primary effector CD8(+) T cell response of substantial magnitude that could be readily quantitated with a simple set of four phenotypic markers. Secondly, the vaccine-induced T cell response was highly specific with minimal bystander effects. Thirdly, virus-specific CD8(+) T cells passed through an obligate effector phase, contracted more than 90% and gradually differentiated into long-lived memory cells. Finally, these memory cells were highly functional and underwent a memory differentiation program distinct from that described for human CD8(+) T cells specific for persistent viruses. These results provide a benchmark for CD8(+) T cell responses induced by two of the most effective vaccines ever developed.


EPR studies of in vivo radical production by 3,3',5,5'-tetrabromobisphenol A (TBBPA) in the Sprague-Dawley rat.
Brominated flame retardants (BFRs) are present in many consumer products ranging from fabrics to plastics and electronics. Wide use of flame retardants can pose an environmental hazard and it is of interest to determine the mechanism of their toxicity. Of all the BFRs, 3,3',5,5'-tetrabromobisphenol A (TBBPA) is produced in the largest volume. Previous studies by Szymanska et al. (2000) have shown that TBBPA is hepatotoxic in rats. We report here that when TBBPA (100 or 600 mg/kg) dissolved in DMSO and alpha-(4-pyridyl-1-oxide)-N-t-butylnitrone (POBN) was administered ip to male Sprague-Dawley rats the POBN/CH(3) spin adduct was detected by electron paramagnetic resonance (EPR) in the bile. When (13)C-DMSO was employed the POBN/C(13)H(3) adduct was observed. Also present in the bile was the 2,6-dibromobenzosemiquinone radical derived from 2,6-dibromohydroquinone, a known metabolite of TBBPA. Reaction of the 2,6-dibromobenzosemiquinone radical with oxygen would generate superoxide from which hydrogen peroxide can form by dismutation. The hydroxyl radical generated via the Fenton reaction from hydrogen peroxide reacts in vivo with DMSO to give the methyl radical which is trapped by POBN. These observations suggest that the hepatotoxicity of TBBPA in rats may be due to the in vivo generation of the hydroxyl radical as a result of redox reactions involving the TBBPA metabolite 2,6-dibromohydroquinone and its corresponding semiquinone radical.


Size Regulation: Big Insights from Little Cells.
Reporting in Cell, Harris and Theriot (2016) use modeling and quantitative imaging to analyze bacterial cell growth and division. By manipulating surface and volume growth rates, the authors provide insight into bacterial cell size regulation and propose that a threshold level of unincorporated cell wall material specifies when cells divide.


Models and Simulations as a Service: Exploring the Use of Galaxy for Delivering Computational Models.
We describe the ways in which Galaxy, a web-based reproducible research platform, can be used for web-based sharing of complex computational models. Galaxy allows users to seamlessly customize and run simulations on cloud computing resources, a concept we refer to as Models and Simulations as a Service (MaSS). To illustrate this application of Galaxy, we have developed a tool suite for simulating a high spatial-resolution model of the cardiac Ca(2+) spark that requires supercomputing resources for execution. We also present tools for simulating models encoded in the SBML and CellML model description languages, thus demonstrating how Galaxy's reproducible research features can be leveraged by existing technologies. Finally, we demonstrate how the Galaxy workflow editor can be used to compose integrative models from constituent submodules. This work represents an important novel approach, to our knowledge, to making computational simulations more accessible to the broader scientific community.


A randomized controlled trial of the efficacy of orally administered fluralaner (Bravecto™) against induced Ixodes holocyclus (Australian paralysis tick) infestations on dogs.
Ixodes holocyclus ticks are a frequently fatal threat to dogs in eastern Australia. These ticks secrete a neurotoxin that can produce an ascending paralysis after 72 h attachment that can lead to death in affected animals. Fluralaner is a potent systemic acaricide with immediate and persistent efficacy for tick control including evidence of 100% efficacy against Ixodes ricinus ticks within 72 h. This study investigated the potential for oral fluralaner administration to control I. holocyclus infestation and the subsequent risk of host paralysis. Healthy Foxhound and Foxhound cross dogs immunized against holocyclotoxin were randomly allocated to receive either a single fluralaner (at least 25 mg/kg) dose or no treatment. All dogs were penned individually and infested with 30 adult unfed female I. holocyclus 1 day before treatment and 14, 28, 42, 56, 70, 84, 112 and 140 days following treatment. Ticks were counted and assessed at 24, 48 and 72 h after the initial fluralaner treatment and after each subsequent infestation. Ticks were not removed at the 24 and 48 h assessments, but were removed after the 72 h assessments. On 112 and 140 days post treatment a new group of untreated control dogs was used. Fluralaner treatment efficacy against I. holocyclus was 100% at 72 h post treatment. Following re-infestations the efficacy remained at 100% at the 72 h assessments for 115 days and reached 95.7% at 143 days. The differences between mean live tick counts on treatment and control groups were significant (P < 0.00l) at all assessment time points for 143 days following treatment. Oral fluralaner treatment can prevent Australian paralysis tick infestations for at least 115 days.


Hematopoietic stem cell transplantation for Shwachman-Diamond syndrome: experience of the French neutropenia registry.
Our objective was to study the outcome of allogeneic hematopoietic stem cell transplantation (HSCT) for Shwachman-Diamond Syndrome (SDS). Among 71 SDS patients included in the French Severe Chronic Neutropenia Registry, 10 received HSCT between 1987 and 2004 in five institutions. The indications were bone marrow failure in five cases, and myelodysplastic syndrome (MDS) or leukemia in five cases. The median follow-up of patients who survived without relapse is 6.9 years (3.1-16.8 years). The conditioning regimen consisted of a busulfan-cyclophosphamide combination (n=6) or total body irradiation plus chemotherapy (n=4). Six patients received stem cells from unrelated donors and four from identical siblings. Engraftment was complete in eight patients and unassessable in two patients. These latter two patients died of infections 32 and 36 days after HSCT, with grade IV graft-versus-host disease and multiorgan dysfunction. A third patient died from an acute respiratory distress syndrome 17 months after HSCT with progressive granulocytic sarcoma. One patient had an MDS relapse 4 months after HSCT and died 10 months later. The overall 5-year event-free survival rate is 60+/-15%. We conclude that HSCT is feasible for patients with SDS who develop bone marrow failure or malignant transformation.


An ensemble method based on uninformative variable elimination and mutual information for spectral multivariate calibration.
Based on the combination of uninformative variable elimination (UVE), bootstrap and mutual information (MI), a simple ensemble algorithm, named ESPLS, is proposed for spectral multivariate calibration (MVC). In ESPLS, those uninformative variables are first removed; and then a preparatory training set is produced by bootstrap, on which a MI spectrum of retained variables is calculated. The variables that exhibit higher MI than a defined threshold form a subspace on which a candidate partial least-squares (PLS) model is constructed. This process is repeated. After a number of candidate models are obtained, a small part of models is picked out to construct an ensemble model by simple/weighted average. Four near/mid-infrared (NIR/MIR) spectral datasets concerning the determination of six components are used to verify the proposed ESPLS. The results indicate that ESPLS is superior to UVEPLS and its combination with MI-based variable selection (SPLS) in terms of both the accuracy and robustness. Besides, from the perspective of end-users, ESPLS does not increase the complexity of a calibration when enhancing its performance.


Drosophila nerve cord culture: a tool for studying neural development.
The culture of explanted neural tissues has been a useful tool for the study of cellular and molecular neurobiology in vertebrates. We have developed a technique for the culture of explanted ventral nerve cords from Drosophila embryos. We have examined the morphology and dynamic behaviour of the growth cones that extend from these nerve cords, and the effect of calcium deprivation on the bundling of axons that are regenerated from the explanted tissue. This technique offers a unique opportunity to combine in vitro techniques for neuronal cell culture with the powerful techniques of genetic analysis that are available with Drosophila.


Drosophila Fed ARA and EPA Yields Eicosanoids, 15S-Hydroxy-5Z,8Z, 11Z, 13E-Eicosatetraenoic Acid, and 15S-Hydroxy-5Z,8Z,11Z,13E,17Z-Eicosapentaenoic Acid.
Drosophila melanogaster has been a widely used as a model system for its powerful genetic tools. However, it remains to be illustrated if Drosophila can be used to examine the biochemical and physiological metabolism of eicosanoids. Thus, the analysis on the metabolism of C20 polyunsaturated fatty acids (PUFA) in Drosophila was implemented with high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Fatty acid (FA) analysis of the whole body, head, and thorax-abdomen in Drosophila showed C20 PUFA could only be found in Drosophila fed diets supplemented with eicosapentaenoic acid (EPA) and arachidonic acid (ARA), but not in Drosophila fed base diets. The C20 PUFA were found in abundance in the head. Drosophila fed ARA- and EPA-supplemented diets yielded 15S-hydroxy-5Z,8Z,11Z,13E-eicosatetraenoic acid [15(S)-HETE] and 15S-hydroxy-5Z,8Z,11Z,13E,17Z-eicosapentaenoic acid [15(S)-HEPE], respectively, while other sampled eicosanoids could not be detected. Similar results were obtained by incubating fly tissue supplemented with ARA or EPA. Furthermore, a genome sequence scan indicated that no gene encoding the key enzymes synthesizing eicosanoids were found in Drosophila. These findings demonstrate that Drosophila may possess a special lipid metabolic system, which is different from mammals.


Chemical properties and biological activity in soils of Mallorca following twenty years of treated wastewater irrigation.
On the Mediterranean island of Mallorca, the use of secondary-treated municipal wastewater in irrigation was introduced with the construction of the first wastewater treatment plants in the 1970s. In this study, the chemical properties and biological activity of 21 arable soils, irrigated for more than 20 years with secondary-treated wastewater, were tested in order to assess their quality. Soil quality was evaluated by measuring cation exchange capacity, pH, calcium carbonate equivalent, soil organic matter, total nitrogen, available phosphorus, water-soluble organic carbon, soil microbial biomass, soil basal respiration, and the activities of the enzymes dehydrogenase, β-glucosidase and alkaline phosphatase. No negative effects of the irrigation treatment were observed on the measured soil parameters. Indeed, soil water-soluble organic carbon, soil microbial biomass and β-glucosidase and alkaline phosphatase activities increased under treated wastewater irrigation. Biological activity of soils irrigated with treated wastewater was affected mainly by soil organic matter content. Although the typical crop management of alfalfa, and other forage crops associated with treated wastewater irrigation, may have contributed to the increase of these parameters, the results suggest that irrigation with treated wastewater is a strategy with many benefits to agricultural land management.


Hydrogen sulfide removal by compost biofiltration: effect of mixing the filter media on operational factors.
The overall goal of this work was to determine the effect of mixing the filter media of a compost biofilter on H(2)S removal efficiency. The behavior of important operational factors such as moisture of filter media, pressure drop and sulfate accumulation were evaluated, considering mixing the media. Additionally, tracer studies were performed in order to determine the effect of mixing the media on gas distribution. H(2)S removal capacity decreased over time, from 100% to 90%. When bed mixing was carried out, the removal capacity remained constant, close to 100%, and moisture content and sulfates accumulation were better controlled at 50% and at 12 mg S-SO(4)/g dry media respectively. In addition, under this operational pattern, an improvement in gas and particle size distribution was observed inside the filter media, fitting the axial dispersion model and the Ergun equation.


Intravenous solid tip lead placement in telemetry implanted dogs. Part 1: Surgical methods, signal quality, and pathological endpoints.
Electrocardiogram (ECG) signals in safety pharmacology studies are generally collected via subcutaneous or epicardial leads. Subcutaneous placement is an easier procedure, but signals often contain artifacts. Epicardial leads offer improved quality but require additional surgical expertise. Signal quality and tolerability of intravenous (IV)/diaphragmatic ECG leads were investigated as a less invasive alternative to the epicardial ECG lead approach for cardiovascular assessment in dogs. Twenty-eight beagle dogs were implanted with PCT (n=14) or PCTP (n=14) transmitters with IV (negative)/diaphragmatic (positive) ECG leads arranged in approximate Lead II configuration. Surgical time for previous epicardial and current IV lead placement approaches was compared. The ECG signals were assessed for up to 32 weeks post-surgery. Signal quality was assessed based on good wave/total wave (GW/TW) ratios calculated using ECG PRO (Ponemah Physiology Platform, Version 4.8) and variability in ECG parameter measurements for each surgical model. Clinical pathology was assessed on all animals before surgery and approximately 2 and 12 weeks post-surgery. A specialized necropsy was conducted on four animals (two PCT and two PCTP) to assess the tolerability of telemetry equipment; selected tissues were examined microscopically. Surgical time using the IV lead method was approximately 18% shorter than the epicardial lead method. The GW/TW ratio for IV lead-implanted dogs indicated good durability of signal that was similar to epicardial leads. Intra- and inter-animal variability in ECG parameter measurements was similar between IV lead-implanted and epicardial lead-implanted dogs. Clinical pathology revealed no noteworthy findings, and the IV/diaphragmatic surgical approach had minimal consequences on local vasculature and associated implantation sites. Advantages of the IV/diaphragmatic lead model include a less invasive and shorter surgical procedure; high tissue tolerance, ECG signal quality, and durability; and data processing procedures similar to that of epicardial leads. Therefore, the IV/diaphragmatic lead configuration is a viable alternative to more invasive surgical approaches for telemetry device implantation in dogs.


Structure and quantitative determination of the major urinary metabolite of thromboxane B2 in the guinea pig.
2,3-Dinor-thromboxane B2 was the major urinary metabolite of thromboxane B2 in the guinea pig. The structure was assessed mainly by mass spectrometric analysis of a number of derivatives of the metabolite and by chemical degradation by oxidative ozonolysis. A method for quantitative determination of 2,3-dinor-thromboxane B2 in guinea pig urine based on multiple ion analysis and octadeuterated 2,3-dinor-thromboxane B2 as internal standard was developed. The basal excretion of the metabolite was 65 +/- 36 (S.D.) ng/kg x 24 h (n = 19; range 19--140 ng). This level corresponded to an endogenous synthesis of 543 +/- 300 ng of TXB2. No increase in the excretion was seen after anaphylaxis, in contrast to what has earlier been reported for PGF2 alpha.


siRNA therapeutics: insights, challenges, remedies and future prospects.
Among conventional and novel therapeutic approaches, the siRNA strategy stands out for treating disease by silencing the gene responsible for the corresponding disorder. Gene silencing is supposedly intended to target any disease-causing gene, and therefore, several attempts and investments were made to exploit siRNA gene therapy and advance it into clinical settings. Despite the remarkable beneficial prospects, the applicability of siRNA therapeutics is very challenging due to various pathophysiological barriers that hamper its target reach, which is the cytosol, and execution of gene silencing action. The present review provides insights into the field of siRNA therapeutics, significant in vivo hurdles that mitigate the target accessibility of siRNA, and remedies to overcome these siRNA delivery challenges. Nonetheless, the current review also highlights the on-going clinical trials and the regulatory aspects of siRNA modalities. The siRNAs have the potential to reach previously untreated target sites and silence the concerned gene owing to their modification as polymeric or lipidic nanoparticles, conjugates, and the application of advanced drug delivery strategies. With such mounting research attempts to improve the delivery of siRNA to target tissue, we might shortly witness revolutionary therapeutic outcomes, new approvals, and clinical implications.


Can MRI biomarkers for hearing loss in enlarged vestibular aqueduct be measured reproducibly?
Morphological features of an enlarged endolymphatic duct (ED) and sac (ES) are imaging biomarkers for genotype and hearing loss phenotype. We determine which biomarkers can be measured in a reproducible manner, facilitating further clinical prediction studies in enlarged vestibular aqueduct hearing loss. A rater reproducibility study. Three consultant radiologists independently measured previously reported MRI ED & ES biomarkers (ED midpoint width, maximal ED diameter closest to the vestibule, ES length, ES width and presence of ES signal heterogeneity) and presence of incomplete partition Type 2 from 80 ears (T2 weighted axial MRI). Interclass correlation coefficients (ICC) and Gwet's Agreement Coefficients (AC) were generated to give a measure of reproducibility for both continuous and categorical feature measures respectively. ES length, width and sac signal heterogeneity showed adequate reproducibility (ICC 95% confidence intervals 0.77-0.95, Gwet's AC for sac heterogeneity 0.64). When determining ED midpoint width, measurements from multiple raters are required for "good" reliability (ICC 95% CI 0.75-0.89). Agreement on the presence of incomplete partition Type 2 ranged from "moderate" to "substantial". Regarding MR imaging, the opinion of multiple expert raters should be sought when determining the presence of an enlarged ED defined by midpoint width. ED midpoint, ES length, width and signal heterogeneity have adequate reproducibility to be further explored as clinical predictors for audiological phenotype. We report which ED & ES biomarkers are reproducibly measured. Researchers can confidently utilise these specific biomarkers when modelling progressive hearing loss associated with enlarged vestibular aqueduct.


Equilibrium partitioning theory to predict the sediment toxicity of the anionic surfactant C(12)-2-LAS to Corophium volutator.
The study of the effect of the sorption of linear alkylbenzene sulfonates (LAS) on the bioavailability to marine benthic organisms is essential to refine the environmental risk assessment of these compounds. According to the equilibrium partitioning theory (EqP), the effect concentration in water-only exposure will be similar to the effect concentration in the sediment pore water. In this work, sorption and desorption experiments with two marine sediments were carried out using the compound C(12)-2-LAS. The effect of the sediment sorption on the toxicity of benthic organisms was studied in water-only and in sediment bioassays with the marine mud shrimp Corophium volutator. In addition, three common spiking methods were tested for its application in the toxicity tests, as well as the stability of the surfactant during the water-only and sediment-water test duration. LC50 values obtained from water-only exposure showed a good correspondence with the pore water concentrations calculated from the sorption and desorption isotherms in the spiked sediments.


Mangiferin depresses vesicular glutamate release in synaptosomes from the rat cerebral cortex by decreasing synapsin I phosphorylation.
Mangiferin is a glucosyl xanthone that has been shown to be a neuroprotective agent against brain disorders involving excess glutamate. However, the effect of mangiferin on the function of the glutamatergic system has not been investigated. In this study, we used synaptosomes from the rat cerebral cortex to investigate the effect of mangiferin on glutamate release and identify the possible underlying mechanism. We observed that mangiferin produced a concentration-dependent reduction in the release of glutamate elicited by 4-aminopyridine with an IC50 value of 25 μM. Inhibition of glutamate release was blocked by removing extracellular calcium and by treatment with the vacuolar-type H+-ATPase inhibitor bafilomycin A1, which prevents the uptake and storage of glutamate in vesicles. Moreover, we showed that mangiferin decreased the 4-aminopyridine-elicited FM1-43 release and synaptotagmin 1 luminal domain antibody (syt1-L ab) uptake from synaptosomes, which correlated with decreased synaptic vesicle exocytosis. Transmission electron microscopy in synaptosomes also showed that mangiferin attenuated the 4-aminopyridine-elicited decrease in the number of synaptic vesicles. In addition, antagonism of Ca2+/calmodulin-dependent kinase II (CaMKII) and protein kinase A (PKA) counteracted mangiferin's effect on glutamate release. Mangiferin also decreased the phosphorylation of CaMKII, PKA, and synapsin I elicited by 4-aminopyridine treatment. Our data suggest that mangiferin reduces PKA and CaMKII activation and synapsin I phosphorylation, which could decrease synaptic vesicle availability and lead to a subsequent reduction in vesicular glutamate release from synaptosomes.


Specific interaction of ERp57 and calnexin determined by NMR spectroscopy and an ER two-hybrid system.
Calnexin and ERp57 act cooperatively to ensure a proper folding of proteins in the endoplasmic reticulum (ER). Calnexin contains two domains: a lectin domain and an extended arm termed the P-domain. ERp57 is a protein disulfide isomerase composed of four thioredoxin-like repeats and a short basic C-terminal tail. Here we show direct interactions between the tip of the calnexin P-domain and the ERp57 basic C-terminus by using NMR and a novel membrane yeast two-hybrid system (MYTHS) for mapping protein interactions of ER proteins. Our results prove that a small peptide derived from the P-domain is active in binding ERp57, and we determine the structure of the bound conformation of the P-domain peptide. The experimental strategy of using the MYTHS two-hybrid system to map interaction sites between ER proteins, together with NMR, provides a powerful new strategy for establishing the function of ER complexes.


Size-dependent specific targeting and efficient gene silencing in peritoneal macrophages using a pH-sensitive cationic liposomal siRNA carrier.
Macrophages are key contributors to various inflammatory diseases. Therefore, the development of an efficient in vivo short interference RNA (siRNA) system that can be delivered to macrophages represents a novel treatment strategy for addressing these disorders. It was recently revealed that peritoneal macrophages (PEMs) are involved in several diseases including ovarian cancer, and are now recognized as a promising drug target. We report herein on the use of pH-sensitive cationic YSK05-MENDs as siRNA carriers and on the impact of both the size of the YSK05-MENDs and their administration routes for the efficient targeting PEMs to achieve a high level of gene silencing activity. The size of the YSK05-MENDs had a dramatic effect on their specificity for PEMs when administered intravenously, but not for intraperitoneal injection. Also, significant gene silencing was achieved by an intraperitoneal administration of the YSK05-MEND at a dose in the single digit μg/kg range. To our knowledge, this is the most efficacious method for siRNA delivery for gene silencing in PEMs in vivo reported to date. These findings enabled us to investigate the complex function of PEMs through several gene silencing simultaneously.


Unusual presentation of neurobrucellosis: a solitary intracranial mass lesion mimicking a cerebral tumor : a case of encephalitis caused by Brucella melitensis.
Among the diverse presentations of neurobrucellosis, solitary intracranial mass lesions are extremely rare. To the best of our knowledge, we describe here the second case of neurobrucellosis mimicking a cerebral tumor caused by Brucella melitensis. The mass lesion was clinically and radiologically indistinguishable from a brain tumor. The diagnosis was established by isolating Brucella melitensis in a blood culture and a positive Wright's agglutination test on the cerebrospinal fluid at 1:320 titers. Paraffin sections of the cerebral mass showed nongranulomatous encephalitis. We suggest that patients with an isolated intraparenchymal mass lesion with nongranulomatous encephalitis should also be studied for brucellosis in endemic areas.


Pathological work-up of sentinel lymph nodes in breast cancer. Review of current data to be considered for the formulation of guidelines.
Controversies and inconsistencies regarding the pathological work-up of sentinel lymph nodes (SNs) led the European Working Group for Breast Screening Pathology (EWGBSP) to review published data and current evidence that can promote the formulation of European guidelines for the pathological work-up of SNs. After an evaluation of the accuracy of SN biopsy as a staging procedure, the yields of different sectioning methods and the immunohistochemical detection of metastatic cells are reviewed. Currently published data do not allow the significance of micrometastases or isolated tumour cells to be established, but it is suggested that approximately 18% of the cases may be associated with further nodal (non-SN) metastases, i.e. approximately 2% of all patients initially staged by SN biopsy. The methods for the intraoperative and molecular assessment of SNs are also surveyed.


Identification of a lupus-susceptibility locus leading to impaired clearance of apoptotic debris on New Zealand Black chromosome 13.
Systemic lupus erythematosus is a chronic multi-organ autoimmune disease marked mainly by the production of anti-nuclear antibodies. Nuclear antigens become accessible to the immune system following apoptosis and defective clearance of apoptotic debris has been shown in several knockout mouse models to promote lupus. However, genetic loci associated with defective clearance are not well defined in spontaneously arising lupus models. We previously showed that introgression of the chromosome 13 interval from lupus-prone New Zealand Black (NZB) mice onto a non-autoimmune B6 genetic background (B6.NZBc13) recapitulated many of the NZB autoimmune phenotypes. Here, we show that B6.NZBc13 mice have impaired clearance of apoptotic debris by peritoneal and tingible-body macrophages and have narrowed down the chromosomal interval of this defect using subcongenic mice with truncated NZB chromosome 13 intervals. This chromosomal region (81-94 Mb) is sufficient to produce polyclonal B- and T-cell activation, and expansion of dendritic cells. To fully recapitulate the autoimmune phenotypes seen in B6.NZBc13 mice, at least one additional locus located in the centromeric portion of the interval is required. Thus, we have identified a novel lupus susceptibility locus on NZB chromosome 13 that is associated with impaired clearance of apoptotic debris.


Characterisation of the murine gene encoding the intracellular hyaluronan receptor IHABP (RHAMM).
We have recently shown that the published cDNA sequence encoding the murine cell surface receptor for hyaluronan-mediated motility (RHAMM) in fact represents a partial sequence of the cDNA encoding a new intracellular hyaluronic acid binding protein (IHABP). Here we publish the genomic organisation, including 700bp sequences of the promoter region, of the IHABP gene. The IHABP gene consists of 18 exons and spans more than 25kb. Part of the IHABP gene is identical with the published data on RHAMM. The IHABP gene apparently possesses one promoter region with one major transcriptional start point. IHABP is ubiquitously expressed at the mRNA and the protein level in all murine tissues, suggesting that the function of this intracellular hyaluronan binding protein is not restricted to migrating cells.


Surgical management of gingival recession: A clinical update.
Gingival recession is defined as the apical migration of the junctional epithelium with exposure of root surfaces. It is a common condition seen in both dentally aware populations and those with limited access to dental care. The etiology of the condition is multifactorial but is commonly associated with underlying alveolar morphology, tooth brushing, mechanical trauma and periodontal disease. Given the high rate of gingival recession defects among the general population, it is imperative that dental practitioners have an understanding of the etiology, complications and the management of the condition. The following review describes the surgical techniques to treat gingival recession.


[Metabolic effects of physical countermeasures against deficient weight-bearing in an experiment with 7-day immersion].
Metabolic effects of physical countermeasures against deficient weight-loading were studied in three groups of 21-30 y.o. volunteers for 7-d dry immersion. Blood serum was investigated for 38 biochemical parameters that characterize myocardium, skeletal musculature, hepatobiliary system, kidney, pancreas, GI tract, prostate, and protein-nucleic, carbohydrate, electrolyte and mineral metabolism. Seven-day DI w/o countermeasures (n = 5) increased concentration of conjugated bilirubin, suppressed activities of muscular (creatine phosphokinase MM) and myocardial enzymes (CPK MB, OBDH), and caused an upward trend in cholesterol, its atherogenic LDP fraction and triglycerides. Mechanic sole stimulation (n = 6) intensified, within the physiological norm, erythrocyte hematolysis raising total bilirubin and potassium. Despite the stimulation, activity of muscle and myocardial enzymes made a decrease. Blood creatinine decreased to a less extent than in the immersed group w/o stimulation, however, lipid parameters did not rise. High-frequency stimulation of the lower leg and hip muscles in the course of immersion (n = 5) was noted to heighten the activity of muscle enzymes and potassium level in blood beyond the physiological norm. Change in creatinine did not reach a statistical significance and lipid metabolism parameters were not different from baseline values. Application of these physical methods of counteracting deficiency of weight bearing did not interfere with redistribution of body liquids due to immersion. Values of the parameters under study were mostly within the normal limits throughout the experimental exposure suggesting absence of pathological developments during DI or in consequence of physical stimulation. Therefore, the reactions were obviously of normal adaptive character.


A comparison of white light laryngostroboscopy versus autofluorescence endoscopy in the evaluation of vocal fold pathology.
To prove the diagnostic value of autofluorescence endoscopy (AFE) and white light laryngostroboscopy (WLS) versus the gold standard microlaryngoscopy with histopathological examination in differential diagnostics of laryngeal lesions for experienced phoniatricians and laryngologists, using the PENTAX SAFE-3000 system. Exploratory cohort study. High-resolution rigid WLS was executed in 32 consecutive patients with initial manifestation of benign, precancerous, and malignant vocal fold lesions. Fiberoptic blue light AFE (SAFE-3000; λ = 408 nm) was subsequently performed by an experienced endoscopist in a blinded study setting. Findings were rated based on objective WLS and AFE parameters (e.g., phonatory vibration, mucosal wave propagation, and loss of autofluorescence). The clinically assumed WLS and AFE diagnoses were compared with the final histopathology of biopsied material taken during microlaryngoscopy. In reference to histopathological diagnosis, WLS achieved a higher sensitivity (100% vs. 94%), specificity (94% vs. 69%), and accuracy (97% vs. 81%) than AFE diagnostics. The concordance between both endoscopic techniques was 87.5% (28/32 patients); additional AFE benefits were not detectable. Significant loss of autofluorescence was observed in malignant findings clinically clearly diagnosed by WLS, but also in chronic inflammation, severe dysplasia, granulomas, vascular polyps, and glottal papillomatosis. The evaluation of vocal fold pathology by the clinically experienced examiner precisely applying WLS appears to be more reliable than diagnostics of mucosal tissue changes by means of AFE via the SAFE-3000 system as a relatively nonspecific method. Microlaryngoscopy with histopathological examination and phonomicrosurgical excision of pathologic changes remains the gold standard.


Landscapes within landscapes: A parasite utilizes different ecological niches on the host landscapes of two host species.
Parasites are distributed across populations of hosts, but also across microhabitats on or inside hosts: together the host population distribution and "host landscape" distribution comprise a part of the ecological niche of a parasite. In this paper we examine how a generalist parasite, the tick Ixodes holocyclus, is distributed at both the host population and host landscape scales in two species of host (Perameles nasuta and Rattus rattus) that co-occur. We anaesthetized wildlife to locate ticks from five generalized host body regions; we then analysed the distribution of ticks among the populations of hosts (aggregation) and the distribution of ticks among the available host body regions as niches. Ixodes holocyclus is more aggregated in the R. rattus population. At the host landscape scale, I. holocyclus's utilized niche includes the entire surface of P. nasuta equally, while the niche on R. rattus is focused on the head. Differences in tick aggregation between host species may reflect tick habitat suitability at the host landscape scale, as well as differences in ecological and evolutionary histories. By exploring the distribution of parasites at multiple scales, including host landscapes, we can better understand the complex ecology of parasites.


Reasons for Low Regulatory Body Discipline Rates for Canadian Hospital Pharmacists.
Past research on disciplinary action by pharmacist regulatory bodies has shown that most cases concern community pharmacists, with few occurring in a hospital setting. To investigate how discipline-related issues involving pharmacists are dealt with by hospital pharmacy departments in Canada. Hospital pharmacy directors and managers from small, medium, and large hospitals across Canada were invited to participate in semi-structured telephone interviews. The interview questions focused on the discipline process in participants' organizations, the situations when reporting to the regulatory body is deemed to be warranted, possible penalties, and recommendations for improving the regulatory body or organizational discipline process. Ten participants, from British Columbia, Saskatchewan, Ontario, New Brunswick, Prince Edward Island, and Newfoundland and Labrador, agreed to be interviewed. Five key themes emerged as contributing to lower rates of hospital pharmacist discipline cases being escalated to the regulatory college level: robust organizational discipline processes independent from the regulatory college, a practice environment promoting competence, union representation, preference for a remedial approach to discipline, and lack of clarity about when to report to the regulatory authority. This study identified a number of reasons why discipline of hospital pharmacists by a regulatory body may be less prevalent than discipline relating to community pharmacists. The main reasons may be lack of clarity about when to report a case to the regulator and a lack of transparency, given that many cases are handled internally within hospitals. Environmental supports for competence and employee protections (e.g., through a union) may also reduce discipline cases.