Structural Characterization of the D179N and D179Y Variants of KPC-2 β-Lactamase: Ω-Loop Destabilization as a Mechanism of Resistance to Ceftazidime-Avibactam.
Klebsiella pneumoniae carbapenemases (KPC-2 and KPC-3) present a global clinical threat, as these β-lactamases confer resistance to carbapenems and oxyimino-cephalosporins. Recent clinically identified KPC variants with substitutions at Ambler position D179, located in the Ω loop, are resistant to the β-lactam/β-lactamase inhibitor combination ceftazidime-avibactam, but susceptible to meropenem-vaborbactam. To gain insights into ceftazidime-avibactam resistance conferred by D179N/Y variants of KPC-2, crystal structures of these variants were determined. The D179N KPC-2 structure revealed that the change of the carboxyl to an amide moiety at position 179 disrupted the salt bridge with R164 present in wild-type KPC-2. Additional interactions were disrupted in the Ω loop, causing a decrease in the melting temperature. Shifts originating from N179 were also transmitted toward the active site, including ∼1-Å shifts of the deacylation water and interacting residue N170. The structure of the D179Y KPC-2 β-lactamase revealed more drastic changes, as this variant exhibited disorder of the Ω loop, with other flanking regions also being disordered. We postulate that the KPC-2 variants can accommodate ceftazidime because the Ω loop is displaced in D179Y or can be more readily displaced in D179N KPC-2. To understand why the β-lactamase inhibitor vaborbactam is less affected by the D179 variants than avibactam, we determined the crystal structure of D179N KPC-2 in complex with vaborbactam, which revealed wild-type KPC-2-like vaborbactam-active site interactions. Overall, the structural results regarding KPC-2 D179 variants revealed various degrees of destabilization of the Ω loop that contribute to ceftazidime-avibactam resistance, possible substrate-assisted catalysis of ceftazidime, and meropenem and meropenem-vaborbactam susceptibility.


Human immunodeficiency virus type 1 subtype C molecular phylogeny: consensus sequence for an AIDS vaccine design?
An evolving dominance of human immunodeficiency virus type 1 subtype C (HIV-1C) in the AIDS epidemic has been associated with a high prevalence of HIV-1C infection in the southern African countries and with an expanding epidemic in India and China. Understanding the molecular phylogeny and genetic diversity of HIV-1C viruses may be important for the design and evaluation of an HIV vaccine for ultimate use in the developing world. In this study we analyzed the phylogenetic relationships (i) between 73 non-recombinant HIV-1C near-full-length genome sequences, including 51 isolates from Botswana; (ii) between HIV-1C consensus sequences that represent different geographic subsets; and (iii) between specific isolates and consensus sequences. Based on the phylogenetic analyses of 73 near-full-length genomes, 16 "lineages" (a term that is used hereafter for discussion purposes and does not imply taxonomic standing) were identified within HIV-1C. The lineages were supported by high bootstrap values in maximum-parsimony and neighbor-joining analyses and were confirmed by the maximum-likelihood method. The nucleotide diversity between the 73 HIV-1C isolates (mean value of 8.93%; range, 2.9 to 11.7%) was significantly higher than the diversity of the samples to the consensus sequence (mean value of 4.86%; range, 3.3 to 7.2%, P < 0.0001). The translated amino acid distances to the consensus sequence were significantly lower than distances between samples within all HIV-1C proteins. The consensus sequences of HIV-1C proteins accompanied by amino acid frequencies were presented (that of Gag is presented in this work; those of Pol, Vif, Vpr, Tat, Rev, Vpu, Env, and Nef are presented elsewhere [http://www.aids.harvard.edu/lab_research/concensus_sequence.htm]). Additionally, in the promoter region three NF-kappa B sites (GGGRNNYYCC) were identified within the consensus sequences of the entire set or any subset of HIV-1C isolates. This study suggests that the consensus sequence approach could overcome the high genetic diversity of HIV-1C and facilitate an AIDS vaccine design, particularly if the assumption that an HIV-1C antigen with a more extensive match to the circulating viruses is likely to be more efficacious is proven in efficacy trials.


Effect of Instructions Emphasizing Velocity or Accuracy Given in a Random or Blocked Order on Performance Testing and Kinematics in Dart Throwing.
The aims of this study were to investigate the effects of throwing instruction (i.e., velocity and accuracy) and testing order (random or blocked) on dart throwing performance testing and on the movement strategies leading to this performance. Twelve physical education students (nine men and three women, age: 24 ± 7.5; mass: 77.7 ± 15.8, height: 1.77 ± 0.06 m) performed dart throws with four different instructions, varying in priority regarding velocity and accuracy, instructed in a blocked and random manner. The main findings were that dart velocity decreased when the priority of accuracy increased. However, when accuracy was the main priority, accuracy increased only when measured for consistency. Testing order influenced peak joint kinematics of wrist flexion in addition to finger extension and the time of occurrence of elbow extension. Instructions emphasizing velocity and/or accuracy showed a clear speed-accuracy trade-off in dart throwing and thereby followed Fitts' law. Testing order had a minor effect on the speed-accuracy trade-off. The blocked testing order appeared to increase performance outcomes that favored the priority set by the instruction in contrast to the random test order. These differences were based upon adjustments of joint movements, which were based upon the knowledge of the previous attempts. These adjustments were visible between the different instructions through changes in the execution timing of peak wrist flexion and elbow and finger extension.


Melatonin inhibits apoptosis and improves the developmental potential of vitrified bovine oocytes.
Vitrification of oocytes has been shown to be closely associated with increased levels of reactive oxygen species (ROS) and apoptotic events. However, little information is available the effect of melatonin on the ROS levels and apoptotic events in vitrified oocytes. Therefore, we studied the effect of melatonin on ROS and apoptotic events in vitrified bovine oocytes by supplementing vitrification solution or in vitro maturation (IVM) and vitrification solution with 10(-9) m melatonin. We analyzed the ROS, mitochondrial Ca(2+) (mCa(2+) ) and membrane potential (ΔΨm), externalization of phosphatidylserine (PS), caspase-3 activation, DNA fragmentation, mRNA expression levels of Bax and Bcl2 l1, and developmental potential of vitrified bovine oocytes. Vitrified bovine oocytes exhibited increased levels of ROS, mCa(2+) , Bax mRNA, and caspase-3 protein and higher rates of PS externalization and DNA fragmentation, and decreased ΔΨm and Bcl2 l1 mRNA expression level. However, melatonin supplementation in vitrification solution or IVM and vitrification solution significantly decreased the levels of ROS, mCa(2+) , Bax mRNA expression, and caspase-3 protein, and PS externalization and DNA fragmentation rates, and increased the ΔΨm and Bcl2 l1 mRNA expression level in vitrified oocytes, resulting in an increased developmental ability of vitrified bovine oocytes after parthenogenetic activation. The developmental ability of vitrified oocytes with melatonin supplementation in IVM and vitrification solution was similar to that of fresh ones. This study showed that supplementing the IVM and vitrification medium or vitrification medium with 10(-9) m melatonin significantly decreased the ROS level and inhibited apoptotic events of vitrified bovine oocytes, consequently increasing their developmental potential.


Age-related changes in vertebral trabecular bone architecture--assessed by a new method.
Cylindrical trabecular bone specimens (d = 7 mm) were drilled in a vertical direction from the central part of the third vertebral body (L3) from 23 normal individuals aged 15-87 years (10 males and 13 females). The bone samples were embedded in methylmetacrylate and sawn in 400 microns thick sections with an arbitrary rotation but a fixed vertical axis. The sections were investigated in polarized light at a magnification of x 8. By using this technique, vertical and horizontal trabeculae were clearly separated due to different colors. Photographs were taken. These were magnified, and trabecular thickness and intertrabecular distance were measured, using a Zeiss-integration plate II. A significant age-related decrease was found in the mean horizontal trabecular thickness (r = -0.71, p less than 0.001), while the mean thickness of the vertical trabeculae was unchanged with age (r = 0.06, n.s.). Furthermore, a significant increase was found both for the mean distance between the horizontal trabeculae (r = 0.79, p less than 0.001) and between the vertical trabeculae (r = 0.75, p less than 0.001). The present study gave a clear and striking visual presentation of both the thinning and disappearance of the horizontal supporting struts in the vertebral trabecular lattice and the total removal of some of the vertical trabeculae--leading to the dramatic loss of bone strength previously demonstrated.


2-Substitution of N6-benzyladenosine-5'-uronamides enhances selectivity for A3 adenosine receptors.
Adenosine derivatives bearing an N6-(3-iodobenzyl) group, reported to enhance the affinity of adenosine-5'-uronamide analogues as agonists at A3 adenosine receptors (J. Med. Chem. 1994, 37, 636-646), were synthesized starting from methyl beta-D-ribofuranoside in 10 steps. Binding affinities at A1 and A2a receptors in rat brain membranes and at cloned rat A3 receptors from stably transfected CHO cells were compared. N6-(3-Iodobenzyl)adenosine was 2-fold selective for A3 vs A1 or A2a receptors; thus it is the first monosubstituted adenosine analogue having any A3 selectivity. The effects of 2-substitution in combination with modifications at the N6- and 5'-positions were explored. 2-Chloro-N6-(3-iodobenzyl)adenosine had a Ki value of 1.4 nM and moderate selectivity for A3 receptors. 2-Chloro-N6-(3-iodobenzyl)adenosine- 5'-N-methyluronamide, which displayed a Ki value of 0.33 nM, was selective for A3 vs A1 and A2a receptors by 2500- and 1400-fold, respectively. It was 46,000-fold selective for A3 receptors vs the Na(+)-independent adenosine transporter, as indicated in displacement of [3H]N6-(4- nitrobenzyl)-thioinosine binding in rat brain membranes. In a functional assay in CHO cells, it inhibited adenylate cyclase via rat A3 receptors with an IC50 of 67 nM. 2-(Methylthio)-N6-(3-iodobenzyl)-adenosine-5'-N-methyluronamide and 2-(methylamino)-N6-(3-iodobenzyl)adenosine-5'-N-methyluronamide were less potent, but nearly as selective for A3 receptors. Thus, 2-substitution (both small and sterically bulky) is well-tolerated at A3 receptors, and its A3 affinity-enhancing effects are additive with effects of uronamides at the 5'-position and a 3-iodobenzyl group at the N6-position.


The effects of non-invasive radiofrequency electric field hyperthermia on biotransport and biodistribution of fluorescent [60]fullerene derivative in a murine orthotopic model of breast adenocarcinoma.
The aim of this study is to understand the combined and differential biokinetic effects of radiofrequency (RF) electric-field hyperthermia as an adjunctive therapy to [60]fullerene nanoparticle-based drug delivery systems in targeting the micro-vasculature and micro-environments of breast cancer tumors. Intravital microscopy (IVM) is an ideal tool to provide the spatial and temporal resolution needed for quantification in this investigation. The water-soluble and fluorescent [60]fullerene derivative (C60-serPF) was designed to be an amphiphilic nanostructure, which is able to cross several biological membranes and accumulate in tumor tissues by passing through abnormally leaky tumor blood vessels. To elucidate the coupled effects of the highly permeable, but heterogeneous tumor vasculature, with the permeabilizing effects of mild (40-42°C) hyperthermia produced by a local RF field, we controlled variables across tumor and non-tumor mammary gland microvasculature with and without application of RF hyperthermia in each condition. We notice that tumor tissue is characterized by more intense drug extravasation than in contralateral mammary fat pad tissue, which is consistent with enhanced permeability and retention (EPR) effects. The analysis of a permeability parameter (Papp), C60-serPF velocity, and the time of compound influx into the intra- and extra-vascular space suggest that mild RF hyperthermia can improve nanoparticle delivery into tumor tissue.


A selective inhibitor of heme biosynthesis in endosymbiotic bacteria elicits antifilarial activity in vitro.
Lymphatic filariasis and onchocerciasis are severe diseases caused by filarial worms and affect more than 150 million people worldwide. Endosymbiotic α-proteobacteria Wolbachia are essential for these parasites throughout their life cycle. Using a high-throughput chemical screen, we identified a benzimidazole compound, wALADin1, that selectively targets the δ-aminolevulinic acid dehydratase (ALAD) of Wolbachia (wALAD) and exhibits macrofilaricidal effects on Wolbachia-containing filarial worms in vitro. wALADin1 is a mixed competitive/noncompetitive inhibitor that interferes with the Mg(2+)-induced activation of wALAD. This mechanism inherently excludes activity against the Zn(2+)-dependent human ortholog and might be translatable to Mg(2+)-responsive orthologs of other bacterial or protozoan pathogens. The specificity profile of wALADin1 derivatives reveals chemical features responsible for inhibitory potency and species selectivity. Our findings validate wALADins as a basis for developing potent leads that meet current requirements for antifilarial drugs.


Promising half-metallicity in ductile NbF3: a first-principles prediction.
Materials with half-metallicity are long desired in spintronics. Using first-principles calculations, we predicted that the already-synthesized NbF3 crystal is a promising half-metal with a large exchange splitting and stable ferromagnetism. The mechanical stability, ductility and softness of the NbF3 crystal were confirmed by its elastic constants and moduli. The Curie temperature (TC = 120 K) estimated from the Monte Carlo simulations based on the 3D Ising model is above the liquid nitrogen temperature (78 K). The ferromagnetism and half-metallicity can be preserved on the surfaces of NbF3. The NbOF2 formed by substituting F with O atoms, however, has an antiferromagnetic ground state and a normal metallic band structure. This work opens an avenue for half-metallic materials and may find applications in spintronic devices.


A Standardized Extract of Lentinula edodes Cultured Mycelium Inhibits Pseudomonas aeruginosa Infectivity Mechanisms.
The priority pathogen list of the World Health Organization classified Pseudomonas aeruginosa as the second top critical pathogen. Hence, the development of novel antibacterial strategies to tackle this bacterium is highly necessary. Herein we explore the potential antibacterial effect of a standardized extract of cultured mycelium of Lentinula edodes (AHCC®) on P. aeruginosa. AHCC® was found to inhibit the growth rate and biofilm formation of strain PAO1. No change in swarming was observed, but AHCC® hampered swimming and twitching motility. In accordance, a decreased expression of metabolism, growth, and biofilm formation genes was shown. AHCC® also diminished the levels of exotoxin A and bacteria inside IEC18 cells and the secretion of IL-6, IL-10 and TNF by infected macrophages. This effect was related to a reduced phosphorylation of MAPKs and to bacteria internalization. Taken together, our data suggest that AHCC® has a potential role to prevent P. aeruginosa infections and may lead to the development of new therapies.


Protective Effect of Water Extract of Citrus Pomace against AAPH-Induced Oxidative Stress In Vitro in Vero Cells and In Vivo in Zebrafish.
Citrus pomace (CP) is a by-product occurred during juice or other products processing. The enormous amount of CP caused serious environmental issues. However, CP is rich in a variety of bioactive compounds. In the present study, a water extract of CP (CPW) was prepared from the by-product and the in vitro and in vivo antioxidant activities of CPW were investigated. The in vitro antioxidant activities of CPW were evaluated by measuring the free radical scavenging activity and protective effects against 2,2-azobis(2-amidinopropane) hydrochloride (AAPH)-induced oxidative stress in Vero cells. CPW scavenges 1,1-diphenyl-2-picrylhydrazyl, alkyl, and hydroxyl radicals at IC50 of 0.16±0.00, 0.31±0.01, and 0.86±0.02 mg/mL, respectively. In addition, CPW improved cell viability and scavenged intracellular reactive oxygen species (ROS) in AAPH-stimulated Vero cells in a dose-dependent manner. The in vivo antioxidant activities of CPW were investigated in a model of AAPH-induced zebrafish embryos. CPW significantly improved the survival rates and reduced heartbeat rates in AAPH-stimulated zebrafish. Furthermore, the intracellular ROS and cell death levels were remarkably decreased in CPW-treated zebrafish. Therefore, the present results indicated that CPW possesses potent in vitro and in vivo antioxidant properties and could be a potential ingredient used in food, pharmaceutical, and cosmetic industries.


Decomposition analysis as a framework for understanding heterogeneity of treatment effects in non-randomized health care studies.
This paper uses the decomposition framework from the economics literature to examine the statistical structure of treatment effects estimated with observational data compared to those estimated from randomized studies. It begins with the estimation of treatment effects using a dummy variable in regression models and then presents the decomposition method from economics which estimates separate regression models for the comparison groups and recovers the treatment effect using bootstrapping methods. This method shows that the overall treatment effect is a weighted average of structural relationships of patient features with outcomes within each treatment arm and differences in the distributions of these features across the arms. In large randomized trials, it is assumed that the distribution of features across arms is very similar. Importantly, randomization not only balances observed features but also unobserved. Applying high dimensional balancing methods such as propensity score matching to the observational data causes the distributional terms of the decomposition model to be eliminated but unobserved features may still not be balanced in the observational data. Finally, a correction for non-random selection into the treatment groups is introduced via a switching regime model. Theoretically, the treatment effect estimates obtained from this model should be the same as those from a randomized trial. However, there are significant challenges in identifying instrumental variables that are necessary for estimating such models. At a minimum, decomposition models are useful tools for understanding the relationship between treatment effects estimated from observational versus randomized data.


Clock1a affects mesoderm development and primitive hematopoiesis by regulating Nodal-Smad3 signaling in the zebrafish embryo.
Circadian clock and Smad2/3/4-mediated Nodal signaling regulate multiple physiological and pathological processes. However, it remains unknown whether Clock directly cross-talks with Nodal signaling and how this would regulate embryonic development. Here we show that Clock1a coordinated mesoderm development and primitive hematopoiesis in zebrafish embryos by directly up-regulating Nodal-Smad3 signaling. We found that Clock1a is expressed both maternally and zygotically throughout early zebrafish development. We also noted that Clock1a alterations produce embryonic defects with shortened body length, lack of the ventral tail fin, or partial defect of the eyes. Clock1a regulates the expression of the mesodermal markers ntl, gsc, and eve1 and of the hematopoietic markers scl, lmo2, and fli1a Biochemical analyses revealed that Clock1a stimulates Nodal signaling by increasing expression of Smad2/3/4. Mechanistically, Clock1a activates the smad3a promoter via its E-box1 element (CAGATG). Taken together, these findings provide mechanistic insight into the role of Clock1a in the regulation of mesoderm development and primitive hematopoiesis via modulation of Nodal-Smad3 signaling and indicate that Smad3a is directly controlled by the circadian clock in zebrafish.


East Indian Sandalwood Oil (EISO) Alleviates Inflammatory and Proliferative Pathologies of Psoriasis.
Psoriasis, a chronic inflammatory skin disease marked by hyper proliferation and aberrant differentiation of keratinocytes, affects 2-3% of the world's population. Research into the pathogenesis of psoriasis has been hampered by the lack of models that accurately reflect the biology of the psoriatic phenotype. We have previously reported that East Indian Sandalwood oil (EISO) has significant anti-inflammatory properties in skin models and hypothesized that EISO might provide therapeutic benefit to psoriasis patients due to its anti-inflammatory and anti-proliferative properties. Here we present interim results from an on-going proof-of-concept Phase 2 clinical trial in which topically applied EISO is demonstrating to be well tolerated and helpful in alleviating mild to moderate psoriasis symptoms. This led us to evaluate the ability of EISO to affect the psoriatic phenotype using MatTek Corporation reconstituted organotypic psoriatic and normal human skin models. EISO had no impact on the phenotype of the normal skin tissue model, however, EISO treatment of the psoriasis tissue model reverted psoriatic pathology as demonstrated by histologic characterization and expression of keratinocyte proliferation markers, Ki67 and psoriasin. These phenotypic affects correlated with suppressed production of ENA-78, IL-6, IL-8, MCP-1, GM-CSF, and IL-1β. Demonstration of the ability of EISO to abrogate these psoriasis symptoms in well-characterized in vitro psoriatic tissue models, supports the hypothesis that the clinically observed symptom alleviation is due to suppression of intrinsic tissue inflammation reactions in afflicted lesions. This study presents a systematic approach to further study the underlying mechanisms that cause psoriasis, and presents data supporting the potential of EISO as a new ethnobotanical therapeutic concept to help direct and accelerate the development of more effective therapies.


Apelin and Apelin Receptor in Follicular Granulosa Cells of Buffalo Ovaries: Expression and Regulation of Steroidogenesis.
Apelin (APLN), as a ligand for APJ (an orphan G-protein-coupled receptor), is an adipokine with pleiotropic effects in many physiological processes of the body. It has an important role in the control of reproduction particularly in females (mainly in control of ovarian function). This study was carried out to investigate the mRNA and protein amounts of APLN/APJ in granulose cells (GCs) of ovarian follicles with small (SF), medium (MF), and large (LF) sizes of buffalo (Bubalus bubalis) and the effect of IGF1 and follicle-stimulating hormone (FSH) on the expression levels of APLN/APJ. In addition, we evaluated the effect of various doses of APLN (isoforms -13 and -17) singly or in combination with IGF1 and FSH on estradiol (E2) and progesterone (P4) secretion in GCs. The mRNA and protein abundance of APLN was the highest in GCs of LF while the APJ expression enhanced with follicle enlargement in GCs (p-value <0.01). IGF1 and FSH elevated the mRNA and protein amounts of APLN and FSH, and IGF1 increased the expression of APJ in buffalo GCs (p-value <0.01). Both isoforms of APLN (-13/-17) singly or in the presence of IGF1 or FSH increased the secretion of E2 and P4 with or without preincubation of cells with APJ antagonist (ML221 10 µM), although we had some variation in the effects. Concurrently, APLN-13/-17 significantly increased the mRNA and protein expression of CYP19A1 and StAR (p-value <0.01). ML221 substantially diminished the secretion of E2 and P4 and also the expression of CY19A1 and StAR in buffalo GCs (p-value <0.01). We also revealed that APLN-13/-17 (10-9 M), singly or in response to IGF1 and FSH, increased the production of E2 and P4 in different times of stimulation. In conclusion, APLN may play a crucial role in steroidogenesis and follicular development in ovarian GCs of buffalo.


Papain in organic solvents: determination of conditions suitable for biocatalysis and the effect on substrate specificity and inhibition.
Synthesis of N-CBZ-(N-Carbobenzoxy)-1-amino-acid methyl esters from N-CBZ-amino acids and methanol has been used as an assay to examine the properties of papain in organic solvents containing small amounts of water. Papain is active in solvents ranging in polarity from acetonitrile to tetrachloromethane. The optimal activity in each solvent varied only about three to four fold, but the amount of added water required to achieve it varied from 4% (v/v) in acetonitrile to 0.05% (v/v) in tetrachloromethane. The enzyme was generally more stable in hydrophobic solvents and at lower water contents. The apparent K(m) value of CBZ-glycine was 26 times higher in acetonitrile than in toluene due to differential partitioning of the substrate between aqueous and organic phases. The substrate specificity of the enzyme was qualitatively little different from that in aqueous solution, with amino acid derivatives still the best substrates. Nitrile analogs of substrates inhibited the enzyme, as they do in aqueous solution, and inhibition by a variety of substituted aromatic hydrocarbons showed that the main specificity of papain for hydrophobic side chains at its S(2) subsite, was little affected. The results show that papain can catalyze reactions under a variety of conditions in organic solvents but its substrate specificity is little changed from that in aqueous media.


An intra-thoracic follicular carcinoma of thyroid: An uncommon presentation.
Follicular carcinoma of thyroid is the second most common type of carcinoma of thyroid, and it may metastasize to bone, lung, brain, and skin. However, the initial presentation of follicular carcinoma of the thyroid as a large intrathoracic mass without any symptoms of thyroid gland enlargement and dysfunction is very rare. We hereby report a case of a 50-year-old male who presented with chief complaints of chest and low back pain. Preliminary evaluation led to the provisional diagnosis of left-sided intrathoracic mass with vertebral metastasis which was suspected to be a case of bronchogenic carcinoma with distant metastasis. Surprisingly, transthoracic biopsy and histopathology revealed metastasis from follicular carcinoma of thyroid. This prompted us for a retrograde evaluation for a primary thyroid malignancy for which an ultrasound and contrast enhanced computed tomography (CECT) of the neck was done which confirmed the presence of a solitary thyroid nodule. Ultrasonography-guided fine-needle aspiration cytology of the nodule revealed follicular carcinoma of thyroid. Histopathological evaluation subsequent to total thyroidectomy revealed follicular carcinoma thyroid, further confirming the diagnosis. The patient was then referred to Department of Nuclear Medicine and Radiotherapy for radionuclide ablation and chemotherapy. We chose to report this case because of its rare presentation as a large intrathoracic mass and the retrograde diagnosis of follicular carcinoma of thyroid. To the best of our knowledge, this is the first report of such a rare case.


Development of Crohn's disease in a patient with multiple sclerosis treated with copaxone.
Copaxone (glatiramer acetate) is a synthetic copolymer mimicking a portion of myelin basic protein, one of several putative autoantigens in multiple sclerosis (MS). Copaxone suppresses the production of tumor necrosis factor (TNF)-alpha, a key mediator of inflammation in MS as well as in other pathologies, such as colitis of interstitial bowel disease (IBD). Copaxone is a drug approved for the treatment of MS, and one that is very well tolerated with a high safety profile and relatively few side effects. Crohn's disease has not been associated with its administration. We describe a patient with MS in remission who had not exhibited any signs of IBD in the past. She had been on Copaxone 20 mg/day treatment for 2 years when she first exhibited gastrointestinal symptoms. Our patient developed Crohn's disease while on Copaxone treatment as a consequence of long-term immunosuppression. Clinicians should be aware that Crohn's disease is a potential novel adverse drug effect of Copaxone.


Nearly exclusive growth of small diameter semiconducting single-wall carbon nanotubes from organic chemistry synthetic end-cap molecules.
The inability to synthesize single-wall carbon nanotubes (SWCNTs) possessing uniform electronic properties and chirality represents the major impediment to their widespread applications. Recently, there is growing interest to explore and synthesize well-defined carbon nanostructures, including fullerenes, short nanotubes, and sidewalls of nanotubes, aiming for controlled synthesis of SWCNTs. One noticeable advantage of such processes is that no metal catalysts are used, and the produced nanotubes will be free of metal contamination. Many of these methods, however, suffer shortcomings of either low yield or poor controllability of nanotube uniformity. Here, we report a brand new approach to achieve high-efficiency metal-free growth of nearly pure SWCNT semiconductors, as supported by extensive spectroscopic characterization, electrical transport measurements, and density functional theory calculations. Our strategy combines bottom-up organic chemistry synthesis with vapor phase epitaxy elongation. We identify a strong correlation between the electronic properties of SWCNTs and their diameters in nanotube growth. This study not only provides material platforms for electronic applications of semiconducting SWCNTs but also contributes to fundamental understanding of the growth mechanism and controlled synthesis of SWCNTs.


Contribution of the PD-L1/PD-1 pathway to T-cell exhaustion: an update on implications for chronic infections and tumor evasion.
Recent clinical data support ideas of Programmed death receptor-ligand 1 (PD-L1; also called B7-H1, CD274) playing an important role in immune evasion of tumor cells. Expression of PD-L1 on tumors strongly correlates with the survival of cancer patients. PD-L1 on tumors interacts with the co-inhibitory molecule Programmed death receptor-1 (PD-1, CD279) on T cells mediating decreased TCR-mediated proliferation and cytokine production. In animal tumor models, blockade of PD-L1/PD-1 interactions resulted in an improved tumor control. In addition, exhausted T cells during chronic viral infections could be revived by PD-L1 blockade. Thus, targeting PD-L1/PD-1 interactions might improve the efficacy of adoptive cell therapies (ACT) of chronic infections as well as cancers. Obstacles for a general blockade of PD-L1 might be its role in mediating peripheral tolerance. This review discusses the currently available data concerning the role of PD-L1 in tumor immune evasion and envisions possibilities for implementation into ACT for cancer patients.


Clinical Application of the PES/WES Index on Natural Teeth: Case Report and Literature Review.
The use of reliable indices to evaluate the aesthetic outcomes in the aesthetic area is an important and objective clinical aid to monitor the results over time. According to the literature various indices were proposed to evaluate aesthetic outcomes of implant-prosthetic rehabilitation of the anterior area like Peri-Implant and Crown Index [PICI], Implant Crown Aesthetic Index [ICAI], Pink Esthetic Score/White Esthetic Score [PES/WES], and Pink Esthetic Score [PES] but none of them was related to prosthetic rehabilitation on natural teeth. The aim of this study is to verify the validity of PES/WES index for natural tooth-prosthetic rehabilitation of the anterior area. As secondary objective, we proposed to evaluate the long-term predictability of this clinical application, one of which is presented below, following the analysis of the most currently accepted literature.


Tough germanium nanoparticles under electrochemical cycling.
Mechanical degradation of the electrode materials during electrochemical cycling remains a serious issue that critically limits the capacity retention and cyclability of rechargeable lithium-ion batteries. Here we report the highly reversible expansion and contraction of germanium nanoparticles under lithiation-delithiation cycling with in situ transmission electron microscopy (TEM). During multiple cycles to the full capacity, the germanium nanoparticles remained robust without any visible cracking despite ∼260% volume changes, in contrast to the size-dependent fracture of silicon nanoparticles upon the first lithiation. The comparative in situ TEM study of fragile silicon nanoparticles suggests that the tough behavior of germanium nanoparticles can be attributed to the weak anisotropy of the lithiation strain at the reaction front. The tough germanium nanoparticles offer substantial potential for the development of durable, high-capacity, and high-rate anodes for advanced lithium-ion batteries.


Transcriptional regulation of stress kinase JNK2 in pro-arrhythmic CaMKIIδ expression in the aged atrium.
c-jun N-terminal kinase (JNK) is a critical stress response kinase that activates in a wide range of physiological and pathological cellular processes. We recently discovered a pivotal role of JNK in the development of atrial arrhythmias in the aged heart, while cardiac CaMKIIδ, another pro-arrhythmic molecule, was also known to enhance atrial arrhythmogenicity. Here, we aimed to reveal a regulatory role of the stress kinase JNK2 isoform on CaMKIIδ expression. Activated JNK2 leads to increased CaMKIIδ protein expression in aged human and mouse atria, evidenced from the reversal of CaMKIIδ up-regulation in JNK2 inhibitor treated wild-type aged mice. This JNK2 action in CaMKIIδ expression was further confirmed in HL-1 myocytes co-infected with AdMKK7D-JNK2, but not when co-infected with AdMKK7D-JNK1. JNK2-specific inhibition (either by a JNK2 inhibitor or overexpression of inactivated dominant-negative JNK2 (JNK2dn) completely attenuated JNK activator anisomycin-induced CaMKIIδ up-regulation in HL-1 myocytes, whereas overexpression of JNK1dn did not. Moreover, up-regulated CaMKIIδ mRNA along with substantially increased phosphorylation of JNK downstream transcription factor c-jun [but not activating transcription factor2 (ATF2)] were exhibited in both aged atria (humans and mice) and transiently JNK activated HL-1 myocytes. Cross-linked chromatin-immunoprecipitation assays (XChIP) revealed that both c-jun and ATF2 were bound to the CaMKIIδ promoter, but significantly increased binding of c-jun only occurred in the presence of anisomycin and JNK inhibition alleviated this anisomycin-elevated c-jun binding. Mutated CaMKII consensus c-jun binding sites impaired its promoter activity. Enhanced transcriptional activity of CaMKIIδ by anisomycin was also completely reversed to the baseline by either JNK2 siRNA or c-jun siRNA knockdown. JNK2 activation up-regulates CaMKIIδ expression in the aged atrium. This JNK2 regulation in CaMKIIδ expression occurs at the transcription level through the JNK downstream transcription factor c-jun. The discovery of this novel molecular mechanism of JNK2-regulated CaMKII expression sheds new light on possible anti-arrhythmia drug development.


A rapid validated UV-HPLC method for the simultaneous determination of the antiretroviral compounds darunavir and raltegravir in their dosage form.
A rapid, simple and sensitive high-performance liquid chromatography (HPLC) method with ultraviolet detection has been developed for quantification of darunavir and raltegravir in their pharmaceutical dosage form. The assay enables the measurement of both drugs with a linear calibration curve (R2= 0.999) over the concentration range 5-100 mg/L. The determination was performed on an analytical Tracer Excel 120 ODSB (15x0.4.6 cm) column at 35ºC. The selected wavelength was 254 nm. The mobile phase was a mixture of 0.037 M sodium dihydrogen phosphate buffer, acetonitrile and methanol (40:50:10, v/v/v) at a flow rate of 2.0 mL/min Nevirapine (50 mg/L) was used as internal standard. Accuracy, intra-day repeatability (n = 5), and inter-day precision (n = 3) were found to be satisfactory, being the accuracy from -4.33 to 3.88% and precisions were intra-day and inter-day, 0.25% and 4.42% respectively in case of darunavir. Raltegravir intra-day and inter-day precisions lower of 1.01 and 2.36%, respectively and accuracy values bet from -4.02 to 1.06%. Determination of the darunavir and raltegravir in their dosage form was done with a maximum deviation of 4%. This analytical method is rapid, easily implantable and offers good results.


Tailoring electronic states of a single molecule using adamantane-based molecular tripods.
Adsorption structures and electronic states of molecular tripods, having a Br atom (BATT) and a ferrocene derivative (Ferrocene-ATT) at the head part of the adamantane-based trithiolate, adsorbed on Au(111) have been investigated using low temperature scanning tunneling microscopy (STM) and scanning tunneling spectroscopy (STS). We found that BATT and Ferrocene-ATT form self-assembled monolayers (SAMs), and their orderings are identical to one another, which suggests that the adsorption structure of adamantane-based molecular tripods is independent of the type of functional substituent attached to the head part. The electronic states originated from the ferrocene group were confirmed in the STS spectrum of Ferrocene-ATT whereas those are absent in the BATT spectrum. We note that the ferrocene part has few interactions with the Au substrate owing not only to the upright geometry of Ferrocene-ATT but also to the insulative properties of the adamantane base. The STS mapping revealed the spatial distribution of the electronic state of Ferrocene-ATT.


Registered Nurses' and Medical Doctors' Experiences of Patient Safety in Health Information Exchange During Interorganizational Care Transitions: A Qualitative Review.
This systematic review aimed to identify, critically appraise, and synthesize the best available literature on registered nurses' and medical doctors' experiences of patient safety in health information exchange (HIE) during interorganizational care transitions. The review was conducted according to the JBI methodology for systematic reviews of qualitative evidence. A total of 5 multidisciplinary databases were searched from January 2010 to September 2020 to identify qualitative or mixed methods studies. The qualitative findings were pooled using JBI SUMARI with the meta-aggregation approach. The final review included 6 original studies. The 53 distinct findings were aggregated into 9 categories, which were further merged into 3 synthesized findings: (1) HIE efficiency and accuracy support patient safety during interorganizational care transitions; (2) inaccuracies in content and structure, along with poor HIE usability, jeopardize patient safety during interorganizational care transitions; and (3) health care professionals' (HCP) actions in HIE are associated with patient safety during interorganizational care transitions. The results of this review identified several advantages of HIE, namely, improvements in patient safety based on reduced human error. Nevertheless, a lack of usability and functionality can amplify the effects of human error and increase the risk of adverse events. In addition, HCPs' individual actions in HIE were found to influence patient safety. Hence, the cognitive and sociotechnical perspectives of work related to HIE should be studied. In addition, HCPs' experiences of each stage of HIE deployment should be clarified to ensure a high standard of patient safety. Registration: PROSPERO CRD42020220631, registered on November 13, 2020.


Continuous theta burst transcranial magnetic stimulation reduces resting state connectivity between visual areas.
Continuous theta burst stimulation (cTBS) is a technique that allows for altering of brain activity. Research to date has focused on the effect of cTBS on the target area, but less is known about its effects on the resting state functional connectivity between different brain regions. We investigated this issue by applying cTBS to the occipital cortex and probing its influence in retinotopically defined regions in early visual cortex using functional MRI. We found that occipital cTBS reliably decreased the resting state functional connectivity (i.e., the correlation of spontaneous activity) between regions of the early visual cortex. In the context of a perceptual task, such an effect could mean that cTBS affects the strength of the perceptual signal, its variability, or both. We investigated this issue in a second experiment in which subjects performed a perceptual discrimination task and indicated their level of certainty on each trial. The results showed that occipital cTBS decreased both subjects' accuracy and confidence. Signal detection modeling suggested that these impairments resulted primarily from a decreased strength of the perceptual signal, with a nonsignificant trend of a decrease in signal variability. We discuss the implications of these experiments for understanding the mechanisms by which cTBS influences brain activity and perceptual processes.


Rituximab for treatment of pachymeningitis in granulomatosis with polyangiitis.
Pachymeningitis is a rare manifestation of granulomatosis with polyangiitis (GPA), and there is a paucity of literature regarding the efficiency of rituximab (RTX) in such cases. We present a woman in her early 50s with GPA who initially responded adequately to conventional therapy. However, the treatment course was complicated by pulmonary cryptococcosis. The patient later developed non-infectious pachymeningitis, which failed to respond to steroids and methotrexate. However, using RTX upfront led to good clinical response and radiological resolution.


Beyond D'Arcy Thompson: Future challenges for quantitative biology.
The centennial of "On Growth and Form" is a good opportunity to reflect on the progress of the quantitative study of living systems and where we would like to see it heading. The era of the physical sciences being a mere vehicle for tool building for biological investigations is over. The approaches taken nowadays are analogous to those that physical scientists have taken within their respective fields for centuries, only that now they ask them about biological phenomena and function. Here I give a brief reflection on where we are and where we should direct our focus next, both from the perspective of the research endeavor as a whole, but also with respect to teaching the next generation of scientists joining the field.


ART and conventional root restorations in elders after 12 months.
Successful use of atraumatic restorative treatment (ART) in children has been reported, but little information is available regarding its use in older adults. The hypothesis of this study was that survival rates of root restorations placed by both ART and the conventional technique were similar. Root-surface caries lesions in 103 institutionalized elders in Hong Kong were treated randomly by either: (1) the conventional approach-caries removed by dental burs, and the cavity filled with light-cured resin-modified glass ionomer; or (2) the ART approach-caries removed by hand instruments, and the cavity filled with chemically cured high-strength glass ionomer. In total, 84 conventional and 78 ART restorations were placed. After 12 months, 63 conventional and 59 ART restorations were reviewed, and the respective 12-month survival rates were 91.7% and 87.0% (p > 0.05). It is concluded that the survival rates of both types of root restorations were high and similar.


Mutations in the nuclear lamin proteins resulting in their aberrant assembly in the cytoplasm.
We have constructed a series of mutations in the human A lamin cDNA to identify and alter the nuclear localization signal using an in vivo functional assay system. The nuclear localization signal in the lamin proteins has both structural and functional similarities with that of the SV40 large T-antigen. Mutations within this functional domain result in the assembly of cytoplasmic tubular structures, and the behavior of these mutants suggests a post-translational dimerization of the lamin proteins prior to their transport into the nucleus. In the course of this work other regions of the carboxy terminus of the A/C lamin proteins have been implicated in the proper assembly and structure of the nuclear envelope.


Lymphocytic adenohypophysitis and lymphocytic infundibuloneurohypophysitis.
One hundred and twenty-four cases of lymphocytic adenohypophysitis have been reported since 1962. Nearly 60% of the cases in women involved inset of the disease in relation to pregnancy. Headache and visual field defects were the most frequent symptoms. Most patients showed signs of either isolated or multiple anterior pituitary hormone deficiency. ACTH secretion was impaired the most frequently, followed by TSH, gonadotropins, GH and PRL secretion. One third of the cases involved hyperprolactinemia. Tissue from patients with lymphocytic infundibuloneurohypophysitis, also suffering from DI, revealed lymphocytic inflammation limited to the infundibulum, stalk, and neurohypophysis. Twenty of these 124 lymphocytic adenohypophysitis patients developed DI before treatment, and neuroimaging studies revealed thickening of the pituitary stalk in some. At least in a few cases, chronic lymphocytic infiltration occurred in both the infundibuloneurohypophysitis and adenohypophysis. Although both lymphocytic adenohypophysitis and infundibuloneurohypophysitis may be caused by autoimmune disorders, the antigens involved may differ.


The Critical Assessment of Oxidative Stress Parameters as Potential Biomarkers of Carbon Monoxide Poisoning.
In conventional clinical toxicology practice, the blood level of carboxyhemoglobin is a biomarker of carbon monoxide (CO) poisoning but does not correspond to the complete clinical picture and the severity of the poisoning. Taking into account articles suggesting the relationship between oxidative stress parameters and CO poisoning, it seems reasonable to consider this topic more broadly, including experimental biochemical data (oxidative stress parameters) and patients poisoned with CO. This article aimed to critically assess oxidative-stress-related parameters as potential biomarkers to evaluate the severity of CO poisoning and their possible role in the decision to treat. The critically set parameters were antioxidative, including catalase, 2,2-diphenyl-1-picryl-hydrazyl, glutathione, thiol and carbonyl groups. Our preliminary studies involved patients (n = 82) admitted to the Toxicology Clinical Department of the University Hospital of Jagiellonian University Medical College (Kraków, Poland) during 2015-2020. The poisoning was diagnosed based on medical history, clinical symptoms, and carboxyhemoglobin blood level. Blood samples for carboxyhemoglobin and antioxidative parameters were collected immediately after admission to the emergency department. To evaluate the severity of the poisoning, the Pach scale was applied. The final analysis included a significant decrease in catalase activity and a reduction in glutathione level in all poisoned patients based on the severity of the Pach scale: I°-III° compared to the control group. It follows from the experimental data that the poisoned patients had a significant increase in level due to thiol groups and the 2,2-diphenyl-1-picryl-hydrazyl radical, with no significant differences according to the severity of poisoning. The catalase-to-glutathione and thiol-to-glutathione ratios showed the most important differences between the poisoned patients and the control group, with a significant increase in the poisoned group. The ratios did not differentiate the severity of the poisoning. The carbonyl level was highest in the control group compared to the poisoned group but was not statistically significant. Our critical assessment shows that using oxidative-stress-related parameters to evaluate the severity of CO poisoning, the outcome, and treatment options is challenging.


[Sjögren's syndrome associated with childhood-onset systemic lupus erythematosus].
Sjögren syndrome (SS) is a common autoimmune disease that exhibits broad organ-specific and systemic manifestations, the most prevalent being decreased lacrimal and salivary gland function, xerostomia, and keratoconjunctivitis sicca. Secondary SS occurs associated with autoimmune diseases, most commonly systemic lupus erythematosus (SLE). Thirty-four childhood-onset SLE patients (2 boys and 32 girls, mean onset age 11.5+/-2.6 years) were evaluated for the presence of secondary SS using the classification criteria for SS revised by Japanese Ministry of Welfare in 1999. Clinical manifestations, serological findings, and renal pathology in SLE patients complicated with SS (SLE+SS, n=14) were compared with those in SLE with no SS autoantibodies (Ro/SS-A or La/SS-B) (SLE-no SS antibodies, n=14). Of all 34 cases, 20 (58.8%) were with positive Ro/SS-A or La/SS-B antibody. Fourteen of the 20 (70.0%) were diagnosed as having secondary SS and all of them were subclinical SS with no sicca symptoms. However, findings of Schirmer test, salivary flow test and minor salivary gland biopsy were positive in 2 of 16 (13.3%), 5 of 16 (31.3%), and 14 of 17 (82.4%) patients, respectively. Compared with the SLE-no SS antibodies group, patients with SLE+SS showed higher level of serum IgG and had higher frequency of anti-U1 RNP antibody and significantly more severe renal involvement. The revised criteria is useful for diagnosis of SS in early stage before the development of exocrine gland damage and appears to be helpful for long term follow-up in childhood-onset SLE associated with SS.


Variations in sequence and occurrence of SSU rDNA group I introns in Monilinia fructicola isolates.
A group I intron of 418 base pairs in the Monilinia fructicola ribosomal small-subunit sequence was characterized. The absence of such an intron in M. laxa and M. fructigena led to a PCR test for M. fructicola identification based on the presence of this intron. The failure to amplify a PCR fragment for some isolates of M. fructicola recently lead to speculation that the intron might not be present always in M. fructicola. In this study, we analyzed 13 isolates of M. fructicola and found that the intron was absent in four isolates and we determined from sequence analysis that there are several nucleotide variations that allow the M. fructicola ribosomal SSU intron to be grouped into 6 polymorphic types.


Sudden Unexpected Death in Epilepsy (SUDEP): A Review of Risk Factors and Possible Interventions in Children.
Sudden unexpected death in epilepsy (SUDEP) is a serious and devastating, yet poorly understood outcome in epilepsy. This review discusses the current knowledge and understanding of SUDEP in children and adolescents. Established risk factors for SUDEP include history of generalized tonic-clonic seizures and nocturnal seizures. Other proposed risk factors include the use of multiple antiseizure medications and poor medication adherence. Possible prevention strategies for SUDEP include improved medication adherence, surgical interventions, nighttime safety, seizure detection devices, and diet. Pediatric providers have a great opportunity to educate families about SUDEP, assess medication adherence, and provide families with tools to improve medication adherence and learn about SUDEP in children and adolescents with epilepsy. Future research in SUDEP aims to further understand the etiology and risk factors of SUDEP, while developing more intervention strategies to prevent SUDEP.


[Neonatal arterial hypertension].
Neonatal arterial hypertension is usually related to an acquired renovascular disease. An increasing number of cases has been reported during the last 10 years, because of the more frequent survival of very sick neonates and the practice of umbilical arterial catheterization. Improved monitoring of arterial pressure in the infant allows an early diagnosis leading to an efficient medical treatment. Available drugs are now accurate to control arterial hypertension and improving vital prognosis. Later outcome claims for long term survey.


Interactive effects of dietary composition and hormonal treatment on reproductive development of cultured female European eel, Anguilla anguilla.
Farmed female eels were fed two experimental diets with similar proximate composition but different n-3 polyunsaturated fatty acid (PUFA) levels. Both diets had similar levels of arachidonic acid (ARA), while levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in one diet were approximately 4.5 and 2.6 times higher compared to the other diet, respectively. After the feeding period, each diet group was divided into two and each half received one of two hormonal treatments using salmon pituitary extract (SPE) for 13 weeks: i) a constant hormone dose of 18.75mg SPE/kg initial body weight (BW) and ii) a variable hormone dosage that increased from 12.5mg SPE/kg initial BW to 25mg SPE/kg initial BW. Results showed a significant interaction between diets and hormonal treatments on gonadosomatic index (GSI), indicating that the effect of broodstock diets on ovarian development depends on both nutritional status and hormonal regime. Females fed with higher levels of n-3 series PUFAs and stimulated with the constant hormonal treatment reached higher GSIs than those receiving the variable hormonal treatment. However, when females were fed lower levels of n-3 series PUFAs there was no difference in the effect of hormonal treatments on GSI. We also found that, independent of hormonal treatment, the diet with higher levels of n-3 series PUFAs led to the most advanced stages of oocyte development, such as germinal vesicle migration. Concentration of sex steroids (E2, T, and 11-KT) in the plasma did not differ between diets and hormonal treatments, but was significantly correlated with ovarian developmental stage. In conclusion, increasing dietary levels of n-3 PUFAs seemed to promote oocyte growth, leading to a more rapid progression of ovarian development in European eel subjected to hormonal treatment.


The treatment of heart failure: the role of neurohumoral activation.
Neurohumoral activation refers to increased activity of the sympathetic nervous system, renin-angiotensin system, vasopressin and atrial natriuretic peptide. It is now known that neurohumoral activation contributes to the transition from ventricular dysfunction to clinical heart failure, and is an independent predictor of poor prognosis in heart failure. Although the treatment of heart failure has traditionally focused on drugs to improve ventricular function, there is increasing evidence that therapeutic modulation of neurohumoral activation is a key to successful treatment of heart failure. For example, there is mounting evidence that angiotensin converting enzyme inhibitors (the unquestioned cornerstone for treatment of heart failure), beta receptor blockers, digitalis, and endurance exercise training exert their benefit in heart failure in large part through neurohumoral modulation. This observation--discussed in this brief review--highlights the concept that compensatory neurohumoral activation to decreased cardiac function may itself contribute to the development of heart failure and its poor prognosis.


Cognitive function of youths born to mothers with opioid and poly-substance abuse problems during pregnancy.
Previous research has provided inconclusive evidence regarding the neuropsychological difficulties of children born to mothers partaking in opioid or poly-drug use during pregnancy. Little is known about how these children fare as they get older. The present longitudinal study includes follow-up data on 45 children born to mothers who used heroin and poly-drugs and a group of 48 children without prenatal drug exposure. Most of the drug-exposed youths were placed in permanent foster or adoptive homes before one year of age. The youths (ages 17 to 21) were administered 10 neuropsychological tests. The drug-exposed youths had cognitive and fine motor functions within the normal range compared to population norms but performed significantly worse than the non-exposed group. There were indications of generally lower cognitive functions rather than specific problems with executive functioning. Lower mean birthweight in the risk group (619 grams mean difference, p < .001) only partially mediated the group differences in cognitive functioning. There was a tendency for youths who had few and early changes in their caregivers or who were born to mothers who had used the least number of different drugs during pregnancy to have the best cognitive scores. The study indicates that youths born to mothers who used multiple drugs during pregnancy are vulnerable relative to their peers within a wide range of cognitive functions. The vulnerability seems to be related not only to the mother's drug use during pregnancy but also to factors such as birthweight and unstable parental care during infancy.


Activation of the alternative complement pathway of guinea-gip by liposomes incorporated with trinitrophenylated phosphatidylethanolamine.
By incorporation of trinitrophenylamino-caproyldipalmitoylphosphatidylethanolamine (TNP-Cap-DPPE) into liposomes composed of an equimolecular mixture of dimyristoylphosphatidylcholine (DMPC) and cholesterol (TNP-Cap-liposomes), liposomes became readily lysed by guinea-pig serum (GPS) in Mg++-EGTA-GVB (gelatin veronal buffered saline containing 2 mM MgCl2 and 10 mM ethyleneglycol-bis (beta-amino-ethyl ether)N-N'-tetraacetate) as well as in GVB++ (gelatin veronal buffered saline containing 0.15 mM CaCl2 and 0.5 mM MgCl2). Since the classical complement pathway (CCP) does not work in Mg++-EGTA-GVB, TNP-Cap-liposome lysis by GPS in Mg++-EGTA-GVB was thought to be mediated by the activation of the alternative complement pathway (ACP). This conclusion was supported by observations that heating of GPS at 50 degrees impaired its lytic activity while C4-deficient GPS was capable of lytic activity, no lysis occurred in EDTA, and there was noted consumption of complement in GPS treated with TNP-Cap-liposomes at 30 degrees. For TNP-Cap-liposome lysis by GPS in Mg++-EGTA-GVB, the epitope density of the TNP hapten was required to be 5% or more of the DMPC. Changing the acyl group of the phosphatidylcholine (PC) significantly influenced the ACP activating capacity of TNP-Cap-liposome. Dipalmitoyl-PC, DMPC and distearoyl-PC facilitated the ACP activating capacity of the TNP-Cap-liposome, while dilauroyl-PC, egg-PC and dioleoyl-PC did not. Furthermore, the length of spacer between TNP and dipalmitoylphosphatidylethanolamine (DPPE) also influenced the ACP activating capacity and maximum activation was noted when the spacer was aminocaproyl. These physicochemical characteristics which increase the ACP activating capacity coincided with those reported to increase the immunogenicity of hapten-sensitized liposomes.


[LUNG CANCER TREATMENT IN 2015: ADVANCES AND FUTURE OUTLOOK].
Every third cancer death in Israel is due to lung cancer. Still, 80% of the lung cancer cases are related to tobacco smoking; and therefore, more must be done for primary prevention, screening for lung cancer among risk cohort and against the tobacco industry. In the last decade, significant progress in the molecular understanding of this disease brought to a marked improvement in the overall survival and in the quality of life of these patients. The treatment paradigm is shifting from "one treatment fits all" regimens to personalized treatment strategies that allow patients to be treated with a variety of novel drugs that prolong their survival and maintain their quality of life. The latest revolutionary concept of immunotherapy in lung cancer is emerging and promises further hope to this population


Prostaglandins E1 and E2 analogues ameliorate mucosal injury secondary to distal colonic obstruction.
After colonic obstruction, increased intraluminal pressure may impair blood circulation and cause mucosal injury that sometimes progresses to perforation. This experimental study aims to evaluated effects of prostaglandin E (PGE) analogues in order to prevent mucosal injury secondary to distal colonic obstruction. This study was carried out on 40 male Wistar rats equally divided into four groups: group 1, control; group 2, colonic obstruction by ligation of the sigmoid colon; group 3, obstruction and administration of PGE1; group 4, obstruction and administration of PGE 2. Mucosal structural changes were assessed and scored by histopathological examination at 24 h postoperatively. The thickness of the mucosal layer of the cecal wall was measured as 179.3 and 122 microm (p <.001) in the control and obstruction alone group, respectively. Mucosal thickness was preserved by PGE1 (170 microm) and PGE2 (157.3 microm) administration. The mean damage score was 0.73 in the control and 3.3 (p <.001) in the obstruction alone group. This score was significantly lower in obstruction groups treated with both PGE1 and PGE2, 1.13 and 1.26 respectively (p <.001). Both PGE1 and PGE2 (PGE1 better than PGE2) ameliorate mucosal injury during distal colonic obstruction. We concluded that the administration of PGE analogues in case of mechanical obstruction of the left colon was beneficial for preserving the structure of the mucosal layer.


The direct actions of cannabidiol and 2-arachidonoyl glycerol at GABAA receptors.
Cannabidiol (CBD) is a major non-intoxicating component of cannabis and possesses anti-epileptic, anxiolytic and anti-hyperalgesic properties. The mechanism of action of CBD in producing such effects remains unclear. Despite evidence that some endogenous and synthetic cannabinoids interact with GABAA receptors, no-one has yet investigated the effects of CBD. Here we used two-electrode voltage clamp electrophysiology to compare the actions of CBD with those of the major central endocannabinoid, 2-arachidonoyl glycerol (2-AG) on human recombinant GABAA receptors (synaptic α1-6βγ2 and extrasynaptic α4β2δ) expressed on Xenopus oocytes. CBD and 2-AG were positive allosteric modulators at α1-6βγ2 receptors, with low micromolar potencies. The maximal level of enhancement seen with either CBD or 2-AG were on α2-containing GABAA receptor subtypes, with approximately a 4-fold enhancement of the GABA EC5 evoked current, more than twice the potentiation seen with other α-subunit receptor combinations. Further we observed β-subunit selectivity, whereby modulatory activity was higher at β2/β3 over β1 subunits. The β1-subunit homologous mutant β2(V436T) substantially diminished the efficacy of both drugs to a third of that obtained with wild-type β2 subunit combinations, but without changing potency. The potency of CBD increased and efficacy preserved in binary α1/α2β2 receptors indicating that their effects do not involve the classic benzodiazepine site. Exploration of extrasynaptic α4β2δ receptors revealed that both compounds enhanced GABA EC5 evoked currents at concentrations ranging from 0.01-1μM. Taken together these results reveal a mode of action of CBD on specifically configured GABAA receptors that may be relevant to the anticonvulsant and anxiolytic effects of the compound.


Lipid metabolism and potentials of biofuel and high added-value oil production in red algae.
Biomass production is currently explored in microalgae, macroalgae and land plants. Microalgal biofuel development has been performed mostly in green algae. In the Japanese tradition, macrophytic red algae such as Pyropia yezoensis and Gelidium crinale have been utilized as food and industrial materials. Researches on the utilization of unicellular red microalgae such as Cyanidioschyzon merolae and Porphyridium purpureum started only quite recently. Red algae have relatively large plastid genomes harboring more than 200 protein-coding genes that support the biosynthetic capacity of the plastid. Engineering the plastid genome is a unique potential of red microalgae. In addition, large-scale growth facilities of P. purpureum have been developed for industrial production of biofuels. C. merolae has been studied as a model alga for cell and molecular biological analyses with its completely determined genomes and transformation techniques. Its acidic and warm habitat makes it easy to grow this alga axenically in large scales. Its potential as a biofuel producer is recently documented under nitrogen-limited conditions. Metabolic pathways of the accumulation of starch and triacylglycerol and the enzymes involved therein are being elucidated. Engineering these regulatory mechanisms will open a possibility of exploiting the full capability of production of biofuel and high added-value oil. In the present review, we will describe the characteristics and potential of these algae as biotechnological seeds.


Adverse renal effects of anaplastic lymphoma kinase inhibitors and the response to alectinib of an ALK+ lung cancer patient with renal dysfunction.
A 62-year-old female patient with renal dysfunction and pulmonary adenocarcinoma developed postoperative recurrence and received carboplatin/pemetrexed and maintenance pemetrexed. As an anaplastic lymphoma kinase (ALK) gene translocation was identified, the therapy was changed to crizotinib. However, the patient's blood creatinine level increased, and her physical status worsened. Alectinib also induced exacerbation of renal dysfunction but was controlled by dose reduction of 140 mg twice daily for 2 weeks treatment and 2 weeks break were repeated, and exhibited a partial response for 16 months. Here, we describe the case in which alectinib treatment had beneficial clinical effects on ALK-positive lung adenocarcinoma, which controlled the adverse renal effects by dose reduction and drug breaks.


Anterior cervicothoracic meningocele treated by cystopleural shunt. Case report and review of the literature.
Anterior meningocele is a rare disease which is commonly observed in the sacral region. Anterio cervicothoracic meningocele is generally diagnosed in adult age and there are few reports in infancy. Different treatment procedures are performed such as ligation, resection and wrapping. The case of a 9-month-old male infant with acute respiratory distress and cyanosis is described. He had also recurrent lung infections for 3 months. The magnetic resonance imaging revealed an anterior cervicothoracic meningocele which was compressing the lung, aorta, esophagus and trachea associated with skeletal abnormalities and C2 - C7 syringohydromyelia. There was no evidence of neurofibromatosis. He was submitted to thoracotomy and treated with cystopleural shunt without valve system. Postoperatively his respiratory distress improved dramatically. Radiologically, the size of meningocele decreased and the syringomyelic cavity disappeared. There was no evidence of recurrence during his followup. Cystopleural shunt procedure is a reliable, simple and effective treatment modality in anterior cervicothoracic meningocele cases. This treatment regimen prevents possible neural damage and also treatments to the associated anomalies such as syringohydromyelia.


Pathological ventricular remodeling: mechanisms: part 1 of 2.
Despite declines in heart failure morbidity and mortality with current therapies, rehospitalization rates remain distressingly high, substantially affecting individuals, society, and the economy. As a result, the need for new therapeutic advances and novel medical devices is urgent. Disease-related left ventricular remodeling is a complex process involving cardiac myocyte growth and death, vascular rarefaction, fibrosis, inflammation, and electrophysiological remodeling. Because these events are highly interrelated, targeting a single molecule or process may not be sufficient. Here, we review molecular and cellular mechanisms governing pathological ventricular remodeling.


Anti-ganglioside antibodies in patients with Guillain Barré syndrome and other neurological disorders.
A study was performed on 59 Guillain-Barré syndrome (GBS) cases, 58 neurological controls (NC) and 60 non-neurological controls (NNC) to investigate the association of anti-ganglioside antibodies in GBS and other neurological disorders. Campylobacter jejuni was isolated from 5.7% of GBS patients. Anti-ganglioside immunoglobulin G was present in 82% and immunoglobulin M in 46% in acute inflammatory demyelinating polyneuropathy patients, 70% and 44% respectively in acute motor axonal neuropathy subgroup and 38% each in acute motor sensory axonal neuropathy subgroup. Though high intensity of anti-gangliosides was present in the GBS patients, the NC patients also had adequate anti-gangliosides compared with the NNC group.


Arthroscopic Management of the Stiff Shoulder.
Shoulder stiffness affects a diverse population of patients suffering a decrease in function and shoulder pain. Arthroscopic management of this debilitating spectrum of pathology is a safe and effective course of action in cases recalcitrant to nonoperative therapy. Arthroscopic management of the stiff shoulder has been reported to be effective in the treatment of stiffness due to adhesive capsulitis, birth palsy, stiffness in the setting of rotator cuff tears, and osteoarthritis in the posttraumatic patient, in the postoperative patient, and in the throwing athlete. Arthroscopic management is most effective in treating the stiff shoulder in the setting of adhesive capsulitis recalcitrant to nonoperative therapy or posttraumatic stiffness. Results are more guarded in the treatment of postoperative stiffness. Excessive force and trauma to the shoulder, including fracture of the humerus, can be avoided with a 360[degrees] capsular release for shoulder stiffness rather than manipulation under anesthesia.


Immunohistochemical localization of tubulin and actin in rat prostate.
Immunohistochemical localization of the cytoskeleton components alpha tubulin and actin in the rat prostate was achieved. Formalin-fixed paraffin sections were prepared and examined by the avidin-biotin-peroxidase-complex method (ABC method). We observed that the cytoplasm of the glandular epithelial cells were stained by alpha tubulin and actin antibodies. The staining intensity for these proteins was decreased following castration, and recovered by testosterone plus 17 beta-estradiol-administration to the castrated rats. Based on these data, we suggest that alpha tubulin and actin in the rat prostate very likely participate in the secretory and metabolic activities of the glandular epithelial cells of the prostate.


[Diffusion of ofloxacin (OFLX) into prostatic tissue in patients with benign prostatic hypertrophy].
Diffusion of ofloxacin (OFLX) into prostatic tissue was studied in 31 patients with benign prostatic hypertrophy. Concentrations of OFLX in the serum and prostatic tissue were measured at scheduled intervals after 200 mg OFLX oral administration. The mean OFLX level in prostatic tissue and tissue/serum ratio at 2 hours, 4 hours and 6 hours was 3.33 +/- 0.96 micrograms/g (1.25 +/- 0.28) in 10 patients, 2.21 +/- 0.55 micrograms/g (0.92 +/- 0.32) in 9 patients and 2.10 +/- 0.99 micrograms/g (1.01 +/- 0.23) in 12 patients, respectively. OFLX levels in prostatic tissue covered the minimum inhibitory concentration for several pathogenic bacteria detected from the infected prostatic fluid. Therefore, OFLX was thought to be a very useful drug for the treatment of bacterial prostatitis and postoperative infection of prostatic surgery.


Endocrine causes of erectile dysfunction.
Erectile dysfunction (ED) is one of the commonest disorders of male sexual function. Penile erection depends on a complex interaction of psychological, neural, vascular and endocrine factors. Testosterone has an important role in both central and peripheral domains of this process. In this article we discuss the role of testosterone in male sexual function and endocrine causes of ED.


Well-Being Is Associated With Local to Remote Cortical Connectivity.
Wellbeing refers to cognitive and emotional appraisal of an individual's life and social functioning, which is of great significance to the quality of life of an individual and society. Previous studies have revealed the neural basis of wellbeing, which mostly focused on human brain morphology or network-level connectivity. However, local-to-remote cortical connectivity, which plays a crucial role in defining the human brain architecture, has not been investigated in wellbeing. To examine whether wellbeing was associated with local-to-remote cortical connectivity, we acquired resting-state images from 60 healthy participants and employed the Mental Health Continuum Short Form to measure wellbeing, including three dimensions, namely, emotional wellbeing, psychological wellbeing, and social wellbeing. Functional homogeneity (ReHo) and seed-based functional connectivity were used to evaluate local-to-remote cortical connectivity in these participants. For local connectivity, our results showed that ReHo in the right orbitofrontal sulcus was significantly positively correlated with psychological wellbeing but negatively correlated with social wellbeing. For remote connectivity, connectivity within the right orbitofrontal cortex and interhemispheric connectivity of the orbitofrontal sulcus were both positively associated with psychological wellbeing; functional connectivity between the right orbitofrontal sulcus and the left postcentral sulcus was positively associated with social wellbeing. Our results showed that wellbeing was indeed associated with local-to-remote cortical connectivity, and our findings supplied a new perspective of distance-related neural mechanisms of wellbeing.


The Body Mass Index of Adolescents Attending Seventh-Day Adventist Schools in Australia: 2001-2012.
We examined the body mass index (BMI) of students attending Seventh-day Adventist (Adventist) schools in Australia in 2001 and 2012. A total of 3069 students attending Adventist schools in Australia responded to a health and lifestyle survey in 2001 (N = 1335) and 2012 (N = 1734). The survey captured self-reported height and weight, demographics (age, sex, year level, religion), and select health behaviors. Compared with national norms, lower rates of overweight and obesity were observed in the study cohort, but higher rates of underweight. There was no change in the mean BMI of the students attending Adventist schools in Australia from 2001 to 2012. Regression analyses indicated that a lower BMI was associated with age, sex, more regularly eating breakfast, consuming less soft drink, and having a regular exercise program. The students reported a high consumption of fruits, vegetables, and whole grains compared with Australian national norms, and 29% claimed to be vegetarian. Students attending Adventist schools appear to have a lower prevalence of overweight and obesity than the secular population, but a higher prevalence of underweight. The mechanisms through which Adventist schools may influence student's BMI warrants further investigation.


Holliday junctions in FLP recombination: resolution by step-arrest mutants of FLP protein.
The FLP "recombinase" of the 2-micron circle yeast plasmid can resolve synthetic FLP site-Holliday junctions. Mutants of the FLP protein that are blocked in recombination but are normal in substrate cleavage can also mediate resolution. The products of resolution by these mutants are almost exclusively nicked molecules with a protein-bound 3' end. There is no significant asymmetry in strand cleavage (top versus bottom) by the mutants in linear or in circular FLP substrates; nor is there a bias in resolution (toward parentals or toward recombinants) of Holliday junctions (corresponding to top- or to bottom-strand exchange) by wild-type FLP. During normal FLP recombination, a small amount of the expected Holliday intermediate can be detected.


Urogenital schistosomiasis and associated determinant factors among senior high school students in the Dutsin-Ma and Safana Local Government Areas of Katsina State, Nigeria.
Human schistosomiasis is a chronic parasitic disease of poverty caused by the cercariae of digenetic trematodes of the genus Schistosoma. The disease is a major source of morbidity and mortality in 77 low- and middle-income countries in the tropics where 700 million people are at risk. In a bid to provide relevant epidemiological information to boost control of urogenital schistosomiasis at the state level in Nigeria, we conducted this study with the aim of investigating the disease's prevalence and intensity, and the determinant factors responsible for its endemicity. Data on risk factors were obtained from a total of 645 students aged 12─25 years using well- designed questionnaires. Samples were collected between 09:45 and 14:00 in universal bottles. Each10μl centrifuged sample was examined for the eggs of S. haematobium using Motic® (Binocular) Light Microscope (model S-10-P) with a x10 objective. Average infection intensity was recorded as number of eggs per 10 ml of urine sample. Survey data were entered into Microsoft Excel 2010 and analyzed using Epi Info™ 7. Associations among variables were determined using the chi square test and bivariate and multivariate logistic regressions. Prevalence of urogenital schistosomiasis was 30.54 % among the study population, with a mean infection intensity of 30.27 eggs/10 ml of urine. Prevalence and average intensity were higher in males (28.37 % and 32.21 eggs/10 ml of urine respectively) than in females (2.17 % and 5 eggs/10 ml of urine respectively). Water contact activities (X (2) = 29.031, P = 0.0000), sex (male) [X (2) = 109.82; P<0.0001], location (Dutsin─Ma) [X (2) = 7.19; P = 0.0073], age group 18-20 (X (2) = 4.819, P = 0.0281), altitude (531─560 m) [X (2) = 6.84, p = 0.0089], fathers doing other brown─collar jobs (X (2) = 8.449, P = 0.0037) and mothers' occupation (X (2) = 9.470, P = 0.0021) were found to be significantly associated with urogenital schistosomiasis. Boys were six times more likely to be infected with the cercariae of S. haematobium compared to girls [AOR (95 % CI): 6.34 (4.89─8.22)]. Dutsin-Ma and Safana were classified as moderate-risk Local Government Areas for urogenital schistosomiasis. The strong association between the disease and mother's occupation is of utmost importance and suggests a promising control measure: that is, directing health education as well as grassroots mass chemotherapeutic intervention with praziquantel at mothers. A good network including treated pipe-borne water, drainage system, and sewage disposal facilities available should be improved upon. Molluscicides should be provided at highly subsidized rate to help control the disease.


Management of bacterial corneal ulcers.
A prospective microbiological study of 48 patients with corneal ulcers due to bacterial infection was performed. Positive cultures of corneal ulcer samples were obtained in 60% of all patients; about half of these patients had received antimicrobial treatment prior to sampling. A relatively high incidence of Staphylococcus epidermidis was isolated from ulcer patients (27%) compared with normal controls (10%). Gram stains of ulcer samples were positive for organisms in only 27% of all patients and were not considered useful in determining initial therapy in this series. We concluded that treatment should be started with a broad combination of antibiotics while awaiting the culture results.


Fungal keratitis in London: microbiological and clinical evaluation.
To review cases of culture-positive fungal keratitis seen at Moorfields Eye Hospital over a 13-year period to January 2007. Isolates were identified retrospectively from laboratory reports. The clinical records were reviewed. The fungal type, risk factors for infection, in vitro sensitivity, and clinical outcome were recorded. There were 66 isolates from 65 patients (men, 53.8%). Forty (60.6%) of the isolates were subspecies of Candida. The average interval from the onset of keratitis to confirmation of fungal infection was 3.4 weeks (median, 1.0 week; range, 0-16 weeks). Prior ocular surface disease (OSD) or a penetrating keratoplasty (PK) was present in 38 (97.4%) patients with Candida infection, and 29 (74.4%) patients with Candida infection were using topical steroid at the time of diagnosis. The principal risk factors for filamentary fungal infection were trauma (8 cases, 30.8%) or cosmetic contact lens wear (8 cases, 30.8%), with OSD or a prior PK each present in 5 (19.2%) cases. The difference in the proportions of risk factors between the 2 fungal groups was statistically significant (P < 0.000). The visual outcome was similar between groups, and at final review, 27 (41.5%) eyes had a visual acuity of < or = 1/60 and 3 (4.6%) eyes were eviscerated. In vitro sensitivity testing showed full or part sensitivity in 100% of 55 isolates tested against econazole, 87.9% of 58 isolates tested against amphotericin, 75% of 40 isolates tested against itraconazole, and 100% of 20 isolates tested against voriconazole. Candida was the principal isolate, usually from eyes with OSD or a prior PK treated with topical steroids.


Marker-trait association analysis of frost tolerance of 672 worldwide pea (Pisum sativum L.) collections.
Frost stress is one of the major abiotic stresses causing seedling death and yield reduction in winter pea. To improve the frost tolerance of pea, field evaluation of frost tolerance was conducted on 672 diverse pea accessions at three locations in Northern China in three growing seasons from 2013 to 2016 and marker-trait association analysis of frost tolerance were performed with 267 informative SSR markers in this study. Sixteen accessions were identified as the most winter-hardy for their ability to survive in all nine field experiments with a mean survival rate of 0.57, ranging from 0.41 to 0.75. Population structure analysis revealed a structured population of two sub-populations plus some admixtures in the 672 accessions. Association analysis detected seven markers that repeatedly had associations with frost tolerance in at least two different environments with two different statistical models. One of the markers is the functional marker EST1109 on LG VI which was predicted to co-localize with a gene involved in the metabolism of glycoproteins in response to chilling stress and may provide a novel mechanism of frost tolerance in pea. These winter-hardy germplasms and frost tolerance associated markers will play a vital role in marker-assisted breeding for winter-hardy pea cultivar.


Predicting Coronary Artery Aneurysms in Kawasaki Disease at a North American Center: An Assessment of Baseline z Scores.
Accurate risk prediction of coronary artery aneurysms (CAAs) in North American children with Kawasaki disease remains a clinical challenge. We sought to determine the predictive utility of baseline coronary dimensions adjusted for body surface area (z scores) for future CAAs in Kawasaki disease and explored the extent to which addition of established Japanese risk scores to baseline coronary artery z scores improved discrimination for CAA development. We explored the relationships of CAA with baseline z scores; with Kobayashi, Sano, Egami, and Harada risk scores; and with the combination of baseline z scores and risk scores. We defined CAA as a maximum z score (zMax) ≥2.5 of the left anterior descending or right coronary artery at 4 to 8 weeks of illness. Of 261 patients, 77 patients (29%) had a baseline zMax ≥2.0. CAAs occurred in 15 patients (6%). CAAs were strongly associated with baseline zMax ≥2.0 versus <2.0 (12 [16%] versus 3 [2%], respectively, P<0.001). Baseline zMax ≥2.0 had a C statistic of 0.77, good sensitivity (80%), and excellent negative predictive value (98%). None of the risk scores alone had adequate discrimination. When high-risk status per the Japanese risk scores was added to models containing baseline zMax ≥2.0, none were significantly better than baseline zMax ≥2.0 alone. In a North American center, baseline zMax ≥2.0 in children with Kawasaki disease demonstrated high predictive utility for later development of CAA. Future studies should validate the utility of our findings.


[Effect of integrative Chinese and western medicine on T-lymphocyte subsets in treating patients with severe acute respiratory syndrome].
To investigate the therapeutic effect of integrative Chinese and western medicine (ICWM) in treating severe acute respiratory syndrome (SARS) and its influence on T-lymphocyte subsets. Comparative study was conducted in 133 SARS inpatients in Beijing Ditan Hospital, who were divided into 3 groups according to the treatment applied, the basic treated group, the low dose steroid group and the high dose steroid group, and all the 3 groups were subdivided into two groups, Chinese herbs and non-Chinese herbs added, respectively. Chinese drugs for clearing-up heat, dispelling dampness, detoxication, removing stasis, supplementing Qi and nourishing Yin were selected according to patients' syndrome and given additionally to all the three ICWM groups. Retrospective analysis for significance test on changes of T-lymphocyte subsets before and after treatment were carried out. T-lymphocyte counts, including CD3+, CD4+, CD8+, lowered in all patients before treatment, but increased significantly after treated for 3 weeks, the increment in all the low dose steroid treated groups was higher than that in the basic treated groups, and that in ICWM groups was higher than that in non-ICWM groups, respectively. Glycocorticosteroid and Chinese herbal medicine treatments could promote the recovery of T-lymphocyte profile, rationally use of them is the effective therapeutic method.


[Shigella boydii strains possessing a new serovar (SM00-27) isolated from diarrhea of overseas travelers in Japan].
Five Shigella strains isolated from stool cultures of five sporadic imported diarrheal cases in Japan during 1999-2001, did not react to any antisera of the established Shigella serovars. These strains had the typical biochemical characteristics of Shigella boydii, and were biochemically identical. All strains were positive in a PCR assay and a cultured-cell invasion test for invasiveness; these indicate that they can cause shigellosis in humans. The results of antigenic analysis revealed that they did not belong to any of the recognized or provisional serovars, and were serologically indistinguishable. Strain SM00-27 is designated as the test strain for this new S. boydii serovar.


A call for the better utilization of physical activity and exercise training in the defense against cardiovascular disease.
Statins, also known as 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, effectively reduce elevated levels of serum LDL-C concentration and in turn lower cardiovascular morbidity and mortality. Regular exercise and physical activity also have significant preventive effects against cardiovascular diseases by simultaneously reducing multiple risk factors. However, statins also produce a number of adverse events, including muscle pain, which increases dramatically in statin users who also exercise, likely limiting the cardiovascular benefits. Most importantly, reduced physical activity participation due to statin-related side effects can cancel out the benefits of the pharmacological treatment. Although exercise training offers more modest benefits compared to pharmacological therapy against traditional risk factors, considering the total impact of exercise on cardiovascular health, it is now evident that this intervention may offer a greater reduction of risks compared to statin therapy alone. However, primary recommendations regarding cardiovascular therapy still center around pharmacological approaches. Thus a new outlook is called for in clinical practice that provides room for physical activity and exercise training, thus lipid targets can be reached by a combined intervention along with improvements in other cardiovascular parameters, such as endothelial function and low-grade inflammation. Databases such as Pubmed and Google Scholar as well as the reference list of the relevant articles were searched to collect information for this opinion article.


The evidence of apelin has the bidirectional effects on feeding regulation in Siberian sturgeon (Acipenser baerii).
Apelin is a peptide, mainly produced in the brain, which participates in several physiologic effects. However, knowledge about the mechanism of appetite regulation in teleosts, including the role of apelin is not well understood. The aim of this study is to explore the effect of feeding status on the expression of apelin mRNA in the whole brain and the effects of injection of apelin on food intake in Siberian sturgeon (Acipenser baerii). In this study, we first cloned the apelin cDNA sequence of the Siberian sturgeon. We obtained a 1046-bp cDNA fragment, including a 237-bp open reading frame (ORF) that encoded 78 amino acids. Apelin was widely distributed in 11 tissues related to feeding regulation, with the highest expression in thewhole brain, followed by the spleen and trunk kidney. In addition, we measured the effects of periprandial (preprandial and postprandial) change, fasting and re-feeding on apelin mRNA expression in whole brain. The level of apelin mRNA was significantly decreased 1h after feeding. The results of the fasting experiment showed that the expression of apelin mRNA in the brain was significantly reduced after 1day of fasting but consistently increased throughout the 15-day food deprivation period. When the 15-day fasted fish were re-fed, apelin mRNA expression in the brain was significantly increased as compared to that of the control. These results suggest that apelin may play a bidirectional role in the regulation of food intake in the Siberian sturgeon. In order to further examine the effect of apelin on feeding regulation in Siberian sturgeons, acute and chronic intraperitoneal (i.p.) injection experiments were performed and food intakes were recorded. Results showed that acute i.p. injection of apelin-13 reduced food intake, however, chronic i.p. injection apelin-13 increased the food intake for 7days in Siberian sturgeons. In conclusion, our results show that apelin has a bidirectional effect on feeding regulation in Siberian sturgeons by acting as a satiety factor in short-term feeding regulation and a starvation factor in long-term feeding regulation.


Identification and characterization of TIFY family genes in Brachypodium distachyon.
The TIFY family is a plant-specific gene family encoding proteins characterized by a conserved TIFY domain. This family encodes four subfamilies of proteins, including ZIM-like (ZML), TIFY, PPD and JASMONATE ZIM-Domain (JAZ) proteins. TIFY proteins play important roles in plant development and stress responses. In this study, 21 BdTIFYs were identified in Brachypodium distachyon through genome-wide analysis, including 15 JAZ and 6 ZML genes. Analysis of the distribution of conserved domains showed that there are three additional domains (CCT domain, GATA domain and Jas domain) in the BdTIFY proteins besides the TIFY domain. Phylogenetic analysis indicated that these 21 proteins were classified into two major groups. Expression profile of BdTIFY genes in response to abiotic stresses and phytohormones was analyzed using quantitative real-time RT-PCR. Among 21 BdTIFY genes, 12 of them were induced by JA treatment, and 4 of them were induced by ABA treatment. Most of BdTIFY genes were responsive to one or more abiotic stresses including drought, salinity, low temperature and heat. Especially, BdTIFY5, 9a, 9b, 10c and 11a were significantly up-regulated by multiple abiotic stresses. These results provided important clues for functional analysis of TIFY family genes in B. distachyon.


Type 1 diabetes mellitus associated with Graves' disease and Vogt-Koyanagi-Harada syndrome.
A 30-year-old woman was diagnosed as having Vogt-Koyanagi-Harada (VKH) syndrome. Her past history was Graves' disease. She was administered 1 g of methylprednisolone for three days, which was followed by oral prednisolone. Four weeks later, she developed hyperglycemic hyperosmolar coma. She was treated with intravenous normal saline and insulin, and glycemic control was improved. She was diagnosed as having type 1 diabetes mellitus (DM) because of positive test results for glutamic acid decarboxylase (GAD) antibodies and islet cell antibodies. Her human leukocyte antigen (HLA) genotypes were DQB1(*)0401 and DRB1(*)0405, which were susceptible genes for VKH syndrome, type 1 DM and Graves' disease. Therefore, we should consider that VKH syndrome might be associated with type 1 DM or Graves' disease.


Trends in uterine rupture in Enugu, Nigeria.
Our objective was to determine the trend of rupture of the gravid uterus at Enugu, Nigeria and to determine any change in pattern of presentation, management and outcome of such patients. The birth register of 4,333 deliveries at the University of Nigeria Teaching Hospital Enugu from January 1997 and December 2000 were reviewed. Forty-one cases of ruptured uterus were identified and analysed. The incidence of uterine rupture was 1 in 106 deliveries with a mean maternal age of 31.2 years. The majority (75.6%) of the patients were multiparous and had some form of antenatal care (61%) with 19.5% of the total booked at the Teaching Hospital. Many (78.1%) of the patients were in labour for 24?hours or less and 22.0% had oxytocin to augment or induce labour. The majority (68.3%) had a previously scarred uterus and many (53.6%) had lower segment ruptures. At laparotomy 31.7% had repair alone, 29.2% had repair with tubal ligation, 22.0% subtotal hysterectomy and 17.1% total hysterectomy. Perinatal mortality was high (87.8%) and maternal mortality rate 48.8 per 1,000 deliveries. Labour in a previously scarred uterus was the most common aetiological factor followed by obstructed labour in a multiparous woman. The incidence of ruptured uterus is still rising at Enugu, Nigeria but maternal mortality, due to uterine rupture continues to fall. The most commonly performed surgery is repair with or without sterilisation rather than hysterectomy.


Distal femoral cut perpendicular to the mechanical axis may induce varus instability in flexion in medial osteoarthritic knees with varus deformity in total knee arthroplasty: a pitfall of the navigation system.
Two factors that influence the external rotation angle of the femoral rotational axis in total knee arthroplasty (TKA) were assessed in 40 medial osteoarthritic knees with varus deformity. First, the anatomic configuration of the femur was assessed using standardized radiographs of the patients' lower extremities before TKA. Second, the degree of medial soft tissue release was assessed during TKA. The radiographs showed that the characteristics of the femur were lateral bowing of the shaft and external rotation of the condyle in the coronal plane. Therefore, when the distal femur is cut perpendicular to the mechanical axis, the cut surface may be in too much of a valgus position. Furthermore, some degree of medial soft tissue release was necessary in all knees. Medial soft tissue release rotates the femur externally in extension in the coronal plane, and it rotates the femur externally around the femoral axis in flexion relative to the tibia. A distal femoral cut in too much of a valgus position and medial soft tissue release induces varus instability in flexion in knees with lateral bowing of the femoral shaft. Anatomic variation such as femoral bowing should be considered when a navigation system is used for TKA because the navigation system shows only the mechanical axis.


Olanzapine/fluoxetine combination in patients with treatment-resistant depression: rapid onset of therapeutic response and its predictive value for subsequent overall response in a pooled analysis of 5 studies.
To characterize response profiles of olanzapine/fluoxetine combination therapy in treatment-resistant depression (TRD) and to investigate predictive relationships of early improvement with olanzapine/fluoxetine combination for subsequent response/remission during the acute phase of treatment. Results were pooled from 5 outpatient studies comparing oral olanzapine/fluoxetine combination, fluoxetine, or olanzapine for a maximum of 8 weeks in patients with TRD who had at least 1 historical antidepressant treatment failure during the current episode and who failed a prospective antidepressant therapy during the study lead-in period. Mean Montgomery-Asberg Depression Rating Scale (MADRS) total and core mood items scores from the 8-week evaluation period were compared across treatment groups. Positive and negative predictive values (PPVs, NPVs) were computed from olanzapine/fluoxetine combination-treated patients demonstrating response and remission based on whether they demonstrated early improvement. Mean olanzapine/fluoxetine combination MADRS score reductions were significantly greater than fluoxetine by week 0.5 and olanzapine by week 1. Significantly more olanzapine/fluoxetine combination patients demonstrated MADRS onset of response compared with fluoxetine and olanzapine patients (P < .001 for both MADRS total and core mood items). In olanzapine/fluoxetine combination patients, 38.1% exhibited MADRS total score response versus 26.9% of fluoxetine patients (P < .001) and 22.2% of olanzapine patients (P < .001). NPVs for MADRS total and core mood items response and remission ranged from 85.7% to 92.1%; PPVs ranged from 29.9% to 45.1%. Olanzapine/fluoxetine combination is superior to fluoxetine and olanzapine in producing early improvement in patients with TRD. The absence of early improvement is highly predictive for overall response failure. clinicaltrials.gov Identifier: NCT00035321.