Effect of maxillary incisors, lower lip, and gingival display relationship on smile attractiveness.
This study aimed to evaluate the effect of the relationship between maxillary incisors and lower lip in conjunction with the maxillary gingival display on perception of smile attractiveness. The frontal intraoral photograph of ideally aligned teeth was modified using image processing software. Twenty different smiles were obtained with a combination of 10 different maxillary incisor edge-lower lip distances and 10 different maxillary gingival display amounts. Attractiveness of these 20 smiles was evaluated by orthodontists (n = 101), dentists (n = 107), and laypersons (n = 105). Two-way ANOVA was used to compare alterations on smile esthetics, and repeated measures ANOVA was used to evaluate the interaction between the variables. Bonferroni-adjusted pairwise comparisons were applied for determining the differences between the rater groups. Maxillary incisor edge-lower lip distance (P < 0.01), maxillary gingival display amount (P < 0.001), and rater group (P < 0.01) had statistically significant influences on perception of smile attractiveness. Orthodontists prefer to see a minimal amount of mandibular incisor exposure and almost full crowns of maxillary central incisors. Dentists and laypersons are more tolerant of mandibular incisor exposure and maxillary central incisor coverage by the upper lip. None of the rater groups preferred the coverage of maxillary incisor edges or maxillary gingival display. Elimination of maxillary gingival display helps to improve smile attractiveness, whereas the coverage of maxillary incisor edges has a negative influence.


Autoimmune epilepsy: findings on MRI and FDG-PET.
Autoimmune epilepsy (AE) is becoming increasingly recognized as a potentially reversible cause of frequent or medically intractable seizures and cognitive deterioration. We describe various presentations of autoimmune encephalopathy which have specifically presented with seizure and describe reported imaging findings. This is organized as a review of the more common autoantibodies which can specifically precipitate seizure according to the intracellular or extracellular location of the targeted antigen. For each antibody, we illustrate their pathophysiology, characteristic clinical presentations with typical effective treatments and prognoses and imaging findings on MRI and PET/CT exams. Parenchymal involvement is variable with the limbic structures typically affected; however, non-limbic cortex, cerebellum, brainstem and basal ganglia can also be involved. In the acute setting, affected regions typically demonstrate T2 hyperintensity with mild mass effect from edema and increased 18F-fludeoxyglucose uptake. Chronically involved parenchyma will often undergo atrophy and demonstrate decreased metabolism; mesial temporal sclerosis is often the end result when the limbic system is involved. Without treatment, long-term effects from AE range from ongoing cognitive dysfunction and refractory seizures to death. Familiarity with AE may prompt appropriate antibody screening, particularly in cases of refractory seizure disorders. Early investigation and proper management of AE cases may help to prevent parenchymal and neurologic deterioration in these patients.


Incongruent changes in heart rate variability and body weight after discontinuing aerobic exercise in patients with schizophrenia.
A bidirectional connection exists between obesity and altered heart rate variability (HRV). Schizophrenia has been associated with a high risk of obesity and decreased vagal modulation. Few studies have examined the link between obesity and HRV in patients with schizophrenia. The aim of this study was to investigate the effects of aerobic exercise on body weight and HRV, and if so, whether these effects could be sustained after discontinuation of exercise training. A total of 18 overweight patients with schizophrenia completed an 8-week moderate-intensity aerobic exercise program conducted twice weekly for 50min. Body weight and heart rate variability were measured at baseline, week 8, and 4weeks after discontinuation of exercise training. Compared with the control group (15 overweight patients with schizophrenia without exercise training), the exercise group had reduced 2.3kg at week 8. Furthermore, the exercise program increased the low frequency, high frequency, and low frequency plus high frequency of HRV. However, after discontinuation of the exercise program for 4weeks, the changes in body weight and the HRV parameters diverged. All of the HRV parameters returned to their baseline values, but no change was seen in the reduced body weight. This suggests that HRV analysis is a more sensitive tool to detect health conditions in patients with schizophrenia. Although exercise is an easy and effective way to prevent and improve health problems, mental health providers might have underestimated the benefits of exercise in daily clinical practice. A regular exercise program should be considered as an essential part of treatment strategies for patients with schizophrenia.


Primary pulmonary hypertension. World Health Organization.
Primary pulmonary hypertension (PPH) is a rare disease. The annual incidence in the West is 1-2 cases per million population per year. A recent WHO symposium in 1998 has produced a consensus on classification, methods of screening, risk assessment and treatment. PPH is a diagnosis of exclusion after all other secondary causes of pulmonary hypertension are ruled out. Current treatment strategy involves acute vasodilator drug trial where positive responders are treated with high dose calcium channel blockers and anticoagulation. Those who do not show positive response may be commenced on intravenous prostacyclin. Surgical treatment is one option for patients with severe PPH or for symptomatic relief. Prognosis in general is very poor.


Subcutaneous versus intraperitoneal insulin in the management of diabetics on CAPD: a review.
Intraperitoneal and subcutaneous routes of administration for diabetics on CAPD were compared. The comparison included: (1) Control of blood glucose concentration: both methods can provide satisfactory glycemic control for most patients. Changing the method of insulin administration is warranted when one method fails. (2) Effect on plasma insulin levels: intraperitoneal administration can produce a plasma insulin profile similar to the normal profile. This is unusual with subcutaneous administration. Consequences of hyperinsulinemia (hyperlipidemia, hypertension) seem, however, to be similar between the two methods of insulin administration. (3) Effect on peritoneal permeability: permeability characteristics are maintained unchanged, usually, with either method after long-term CAPD. However, insulin is mitogenic in vitro. Theoretically, intraperitoneal insulin could lead to peritoneal fibrosis. (4) Effect on infectious complications of CAPD: a difference in the rate of peritonitis or overall PD catheter-related infections has not been convincingly demonstrated between the two methods of insulin administration. Exit site and tunnel infections with staphylococcus aureus may be more frequent in diabetics receiving insulin subcutaneously. (5) Effect on hepatic structure and function: subcapsular hepatic steatosis was described in diabetics receiving insulin intraperitoneally. The clinical significance of this finding remains to be demonstrated. We conclude that both methods can be applied for insulin administration in diabetics on CAPD. The intraperitoneal method should be tried first in most instances. Prospective studies comparing the two methods are needed.


Systemic spread of an RNA insect virus in plants expressing plant viral movement protein genes.
Flock house virus (FHV), a single-stranded RNA insect virus, has previously been reported to cross the kingdom barrier and replicate in barley protoplasts and in inoculated leaves of several plant species [Selling, B. H., Allison, R. F. & Kaesberg, P. (1990) Proc. Natl. Acad. Sci. USA 87, 434-438]. There was no systemic movement of FHV in plants. We tested the ability of movement proteins (MPs) of plant viruses to provide movement functions and cause systemic spread of FHV in plants. We compared the growth of FHV in leaves of nontransgenic and transgenic plants expressing the MP of tobacco mosaic virus or red clover necrotic mosaic virus (RCNMV). Both MPs mobilized cell-to-cell and systemic movement of FHV in Nicotiana benthamiana plants. The yield of FHV was more than 100-fold higher in the inoculated leaves of transgenic plants than in the inoculated leaves of nontransgenic plants. In addition, FHV accumulated in the noninoculated upper leaves of both MP-transgenic plants. RCNMV MP was more efficient in mobilizing FHV to noninoculated upper leaves. We also report here that FHV replicates in inoculated leaves of six additional plant species: alfalfa, Arabidopsis, Brassica, cucumber, maize, and rice. Our results demonstrate that plant viral MPs cause cell-to-cell and long-distance movement of an animal virus in plants and offer approaches to the study of the evolution of viruses and mechanisms governing mRNA trafficking in plants as well as to the development of promising vectors for transient expression of foreign genes in plants.


CYP1A1m2 polymorphisms regulate estrogen and interleukin-6 in lung cancer.
Lung cancer, the most common cause of cancer-related death in men and women, is responsible for 1.3 million deaths worldwide annually. Women are diagnosed to a greater extent than men with adenocarcinoma and small cell carcinoma, both of which are secretory-type tumors. Never smokers diagnosed with lung cancer are also predominantly female, demonstrating the association of genetic factors with lung carcinogenesis. Several epidemiologic studies have associates certain CYP1A1 genotypes, alone or in combination, with an increased risk of estrogen-related cancer. The aim of this study was to investigate the impact of the CYP and GST polymorphisms along with estrogen and interleukin-6 (IL-6) levels on the risk of lung cancer. Eighty-six lung cancer patients and 60 controls were included in the study. A significantly higher frequency of polymorphisms in the genes was observed in lung cancer patients compared to controls. Mean estradiol concentration was reduced and IL-6 levels were elevated in patients compared to controls. In conclusion, increased polymorphisms in metabolic genes may be the reason for the reduced estradiol and, thereby, the increased expression of IL-6 in the serum of lung cancer patients.


Fasudil, a Rho kinase inhibitor, suppresses tumor growth by inducing CXCL14/BRAK in head and neck squamous cell carcinoma.
CXCL14/BRAK (BRAK) is a secreted chemokine with anti-tumor activity, and its expression is suppressed in tumor cells. We previously reported the anti-tumor activity of BRAK in cell lines of head and neck squamous cell carcinoma (HNSCC) and the suppression of BRAK secretion in these cells. BRAK secretion in fibrosarcoma cells is restored by Fasudil, which is a Rho-kinase (ROCK) inhibitor. In this study, we examined the anti-tumor effect of BRAK by evaluating its gene expression and protein secretion in HNSCC cell lines. We found that BRAK mediated the suppressive effect of Fasudil against HNSCC cells. Tumor development in female BALB/cAJclnu/nu mice was suppressed by Fasudil. Also secretion of BRAK protein by tumor cell lines in vitro was significantly stimulated by Fasudil treatment. Similarly, the production of BRAK protein was significantly increased by the addition of Fasudil to cultured tumor cells. Furthermore Fasudil significantly increased BRAK gene expression at the mRNA level in HNSCC cell line. Inhibition of the RhoA/ROCK pathway by siRNAs significantly stimulated BRAK gene expression. These results show that the tumor-suppressive effect of Fasudil was mediated by BRAK, suggesting that Fasudil may therefore be useful for the treatment of HNSCC.


Characterization of domestic graywater and graywater solids.
The knowledge of loads and concentrations is fundamental for the design of graywater treatment units, but the data on the characteristics of graywater and in particular graywater solids are weak. As general design values regarding graywater treatment facilities are not available for Germany, the objective of this article is to elaborate the characteristics of graywater and graywater solids. This paper describes the results of six sampling campaigns carried out on graywater systems in the German cities Berlin, Lübeck and Kiel. All graywater samples were collected proportional to the flow and the graywater solids were gathered separately. The collected data include graywater volumes and characteristics regarding the organic pollution (chemical oxygen demand (COD), 5-day biochemical oxygen demand (BOD5)) and nutrients (total nitrogen (TN), total phosphorus (TP)). The graywater volume fluctuated depending on the location. The specific average flow was 68 litre per inhabitant per day (L/inh.d). Inhabitant-specific loads of 49.3 gCODt/inh·d, 28 gBOD5/inh.d, 1 gTNt/inh.d and 0.38 gTPt/inh.d (subscript 't' = total) were found. Information about the composition of graywater solids in terms of quantity and quality is seriously lacking. Therefore, graywater solids were examined with respect to organic matter (COD) and nutrients (TN, TP). The contribution of graywater solids with particle sizes over 200 microns in relation to the total inhabitant-specific load was approximately 3-8% depending on the parameter. The qualitative and quantitative characteristics of the investigated graywater fractions may serve as a base for the estimation of design values.


Combined immunoglobulin and azathioprine in multiple sclerosis.
In an attempt to prevent exacerbations of multiple sclerosis, immunoglobulin therapy was combined with azathioprine (AZA). Intravenous immunoglobulin (i.v.IG) 2 g/kg was given in divided doses over 3 consecutive days followed by monthly booster doses (0.2 g/kg) for 3 years to 38 patients with relapsing-remitting multiple sclerosis (MS). In the 34 patients who completed the trial, the relapse rate decreased (from 1.7 relapses per year to 0 during the 3-year trial period). The Kurtzke Expanded Disability Status Scale decreased from 3.4 +/- 0.72 to 3.0 +/- 0.70. The results suggest that combined i.v.IG and AZA suppress the ongoing pathologic process in relapsing-remitting MS.


Detection of mitochondrial DNA mutations by high-throughput sequencing in the blood of breast cancer patients.
Mitochondrial DNA mutations have been identified in serveral types of cancer. In breast cancer, germline and somatic mitochondrial DNA (mtDNA) mutations have been identified. A number of mtDNA mutations in breast cancer have been identified in protein-coding regions (in protein-coding genes, such as ND2, COX3, ND4, ND5 and CytB). Mutations in these structure proteins cause impaired electron transport function and lead to electron leakage and increased reactive oxygen species (ROS) production, which in turn increases oxidative stress and oxidative damage to the mitochondria, as well as to cells. These data establish an association between mtDNA mutations and breast cancer; however, there is no reliable prediction of breast cancer predisposition or progression based on mtDNA mutation patterns thus far. In this study, we used high-throughput sequencing to detect mtDNA mutations in the blood of breast cancer patients. Some of these mutations may be used as potential markers for breast cancer diagnosis.


MiR-20a regulates fibroblast-like synoviocyte proliferation and apoptosis in rheumatoid arthritis.
STAT3 expression is elevated in the synovial tissue of patients with rheumatoid arthritis (RA). MiR-20a plays a role in mediating synovial inflammation in RA. Bioinformatics analysis has identified a binding site between miR-20 and the 3'-UTR of STAT3 mRNA. This study aimed to investigate the role of miR-20a in the regulation of STAT3 expression and synovial cell proliferation as well as apoptosis. Synovial tissues were collected from RA patients and osteoarthritis (OA) patients to measure miR-20a, STAT3, p-STAT3, and Ki-67 expressions. Fibroblast-like synoviocytes (FLS) were treated with IL-17 (10 ng/ml) and then Ki-67 expression and cell cycle were evaluated by flow cytometry. The targeting relationship between miR-20a and STAT3 was assessed by dual luciferase reporter gene assay. FLS cells were divided into five groups: miR-NC, miR-20a mimic, si-NC, si-STAT3, and miR-20a mimic + si-STAT3 groups. In RA patients, significantly lower MiR-20a expression, and substantially higher STAT3, p-STAT3, and Ki-67 expression were found in the synovial tissues compared with those in OA patients. IL-17A treatment markedly promoted FLS cell proliferation, inhibited cell apoptosis, reduced miR-20a expression, as well as upregulated levels of STAT3, p-STAT3, and Bcl-2. MiR-20a played a regulatory function on the expression of STAT3. MiR-20a mimic and/or si-STAT3 transfection apparently downregulated STAT3, p-STAT3, and Bcl-2 expression, attenuated IL-17A-induced cell proliferation promotive and enhanced cell apoptosis in FLS cells. The expression of miR-20a was reduced in synovial tissue of RA patients with the increased level of STAT3. Downregulation of miR-20a promoted the expression of STAT3, p-STAT3, and Bcl-2, facilitated FLS cell proliferation, reduced apoptosis and, thereby, played a critical role in RA.


Biophysical characterization of interactions between the C-termini of peripheral nerve claudins and the PDZ₁ domain of zonula occludens.
Our recent study has shown that cellular junctions in myelin and in the epi-/perineruium that encase nerve fibers regulate the permeability of the peripheral nerves. This permeability may affect propagation of the action potential. Direct interactions between the PDZ₁ domain of zonula occludens (ZO₁ or ZO₂) and the C-termini of claudins are known to be crucial for the formation of tight junctions. Using the purified PDZ₁ domain of ZO₂ and a variety of C-terminal mutants of peripheral nerve claudins (claudin-1, claudin-2, claudin-3, claudin-5 in epi-/perineurium; claudin-19 in myelin), we have utilized NMR spectroscopy to determine specific roles of the 3 C-terminal claudin residues (position -2, -1, 0) for their interactions with PDZ₁ of ZO₂. In contrast to the canonical model that emphasizes the importance of residues at the -2 and 0 positions, our results demonstrate that, for peripheral nerve claudins, the residue at position -1 plays a critical role in association with PDZ₁, while the side-chain of residue 0 plays a significant but lesser role. Surprisingly, claudin-19, the most abundant claudin in myelin, exhibited no binding to ZO₂. These findings reveal that the binding mechanism of claudin/ZO in epi-/perineurium is distinct from the canonical interactions between non-ZO PDZ-containing proteins with their ligands. This observation provides the molecular basis for a strategy to develop drugs that target tight junctions in the epi-/perineurium of peripheral nerves.


Cluster headache in women: clinical characteristics and comparison with cluster headache in men.
To study the clinical characteristics of cluster headache in women. Cluster headache is a disorder of men (male to female ratio 6-7:1). Retrospective chart review to identify all women diagnosed with cluster headache at an academic headache centre from January 1995 through July 1998. Thirty two women and 69 men were identified. The mean age of onset of cluster headache was 29.4 years in women versus 31.3 years in men. Two peaks of onset in women (2nd and 5th decade) were identified compared with one in men (3rd decade). Episodic cluster headache was present in 75% of women and 77% of men. Women and men had on average 3 attacks a day, but attack duration was shorter in women (67.2 minutes v 88.2 minutes). Cluster headache period duration (11.1 weeks v 10 weeks) and remission periods (21.1 months v 23.1 months) were similar in women and men. Miosis and ptosis seemed to be less common in women (miosis 13.3% v 24.6%, ptosis 41.9% v 58.1%) whereas lacrimation and nasal congestion/rhinorrhoea were almost equally prevalent in women and men. Women had more nausea than men (62.5% v 43.5%, p=0.09) and significantly more vomiting (46.9% v 17.4%, p=0.003). Photophobia occurred in 75% of women and 81.2% of men, and phonophobia occurred in 50% of women and 47.8% of men. The clinical characteristics of cluster headache in women are very similar to those in men. Women develop the disorder at an earlier age of onset and experience more "migrainous symptoms" with cluster headache, especially vomiting. Both men and women have frequent photophobia and phonophobia with cluster headache attacks. These symptoms are not included in the International Headache Society cluster headache criteria, suggesting the need for possible criteria revision.


[Ultrastructural and immunohistochemical analysis of cells during the changeover period in isoenzyme synthesis during drosophila ontogenesis].
The ultrastructural and immunohistochemical studies of the differentiating epithelial cells of the ejaculatory bulb in D. littoralis Maigen (=D. imeretensis Sokolov) have shown that the change of synthesis of non-specific isozyme of carboxyl esterase to synthesis of organospecific carboxyl esterase is realized at the same time in all epithelial cells. The epithelium of ejaculatroy bulb is homogeneous by its celluaar composition at all stages of differetiation under study. No mitosing cells or distinct foci of degeneration were found in the epithelial layer after the flight of imago.


Copper intoxication inhibits aerobic nucleotide synthesis in Streptococcus pneumoniae.
Copper is universally toxic in excess, a feature exploited by the human immune system to facilitate bacterial clearance. The mechanism of copper intoxication remains unknown for many bacterial species. Here, we demonstrate that copper toxicity in Streptococcus pneumoniae is independent from oxidative stress but, rather, is the result of copper inhibiting the aerobic dNTP biosynthetic pathway. Furthermore, we show that copper-intoxicated S. pneumoniae is rescued by manganese, which is an essential metal in the aerobic nucleotide synthesis pathway. These data provide insight into new targets to enhance copper-mediated toxicity during bacterial clearance.


Site-specific incorporation of a redox-active amino acid into proteins.
The redox-active amino acid 3,4-dihydroxy-l-phenylalanine (DHP), which can undergo two-electron oxidation to a quinone, has been incorporated selectively and efficiently into proteins in Escherichia coli in response to a TAG codon. We have demonstrated that DHP can be oxidized electrochemically within the protein. The ability to incorporate a redox-active amino acid site specifically into proteins should facilitate the study of electron transfer in proteins, as well as enable the engineering of redox proteins with novel properties.


Population genetics and morphological comparisons of migratory European (Hirundo rustica rustica) and sedentary East-Mediterranean (Hirundo rustica transitiva) barn swallows.
Speciation processes are largely determined by the relative strength of divergent selection versus the magnitude of gene flow. The barn swallow (Hirundo rustica) has a broad geographic distribution that encompasses substantial geographic variation in morphology and behavior. The European (H. r. rustica) and East-Mediterranean (H. r. transitiva) subspecies are closely related, despite differing in morphological and life-history traits. To explore patterns of genetic differentiation and gene flow, we compared morphological and genetic variation among the nonmigratory breeding population of H. r. transitiva from Israel and the migratory population of H. r. rustica that passes through Israel and compared it with the genetic differentiation between H. r. transitiva from Israel and a breeding population of H. r. rustica from the United Kingdom that uses a different migratory flyway. Mitochondrial haplotype network analysis suggests that the European and East-Mediterranean populations are intermixed, although there was low but significant genetic differentiation between the subspecies based on both mitochondrial (F(ST) = 0.025-0.033) and microsatellite (F(ST) = 0.009-0.014) loci. Coalescent-based analyses suggest recent divergence and substantial gene flow between these populations despite their differences in morphological and behavioral traits. The results suggest that these subspecies are undergoing a differentiation process in the face of gene flow, with selection possibly operating on sexually selected traits.


A clinical Trial of Tofranil in osteo-arthritis.
Animal experiments and uncontrolled clinical experience indicate that imipramine should relieve pain or enhance the effect of analgesics in osteoarthritis. A double-blind crossover trial using Tofranil 25 mg three times daily and matching placebo has, therefore, been designed and initiated.


Unconventional Synthesis of Hierarchically Twinned Copper as Efficient Electrocatalyst for Nitrate-Ammonia Conversion.
Twin boundary (TB) engineering provides exciting opportunities to tune the performance levels of metal-based electrocatalysts. However, the controllable construction of TB greatly relies on surfactants, blocking active sites and electron transfer by surfactants. Here, we propose a surfactant-free and facile strategy for synthesizing copper (Cu) nanocatalysts with dense hierarchical TB networks (HTBs) by the rapid thermal reductions in metastable CuO nanosheets in H2 . As revealed by in situ transmission electron microscopy (TEM), the formation of HTBs is associated with the fragmentation of nanosheets in different directions to generate abundant crystal nuclei and subsequently unconventional crystal growth through the collision and coalescence of nuclei. Impressively, the HTBs endow Cu with excellent electrocatalytic performance for direct nitrate-ammonia conversion, superior to that of Cu with a single-oriented TB and without TB. We discover that the HTBs induce the formation of compressive strains, thereby creating a synergistic effect of TBs and strains to efficiently tune the binding energies of Cu with nitrogen intermediates (i.e., NO2 *) and thus promote the tandem reaction process of NO3 - -to-NO2 - and subsequent NO2 - -to-NH3 electrocatalysis. This work demonstrates the crucial role of HTBs for boosting electrocatalysis via the synergistic effect of TBs and strains. This article is protected by copyright. All rights reserved.


Oxidative degradation of lipids during mashing.
Although hardly any polyunsaturated fatty acids (PUFAs) are present in the endproduct, the ingredients used for the production of beer contain a high concentration of PUFAs, such as linolic and linolenic acid. These compounds are readily oxidized, resulting in the formation of lipid-derived products that reduce the taste and quality of beer enormously. During mashing relatively high amounts of PUFAs are exposed to atmospheric oxygen at a relatively high temperature. This makes mashing a critical step in the brewing process with regard to the formation of lipid-derived off-taste products. F1 phytoprostane (PPF1) changes in antioxidant capacity and monohydroxy fatty acids (OH-FAs) were used as markers for the detection of oxidative damage to fatty acids during mashing. The pattern of OH-FA formation indicates that enzymatic oxidation of PUFAs is more important than nonenzymatic oxidation during the mashing process. Nevertheless, substantial nonenzymatic radical formation is evident from the increase of specific OH-FAs and PPF1s. It was found that a low oxygen tension reduces oxidative damage and gives a high antioxidant capacity of the mashing mixture. This indicates that mashing should be done under low oxygen pressure.


Changes in mitochondrial membrane potential and reactive oxygen species during wogonin-induced cell death in human hepatoma cells.
Flavonoids exist extensively in plants, and several biological effects of them have been demonstrated. Wogonin is an important flavonoid compound. In this study, wogonin showed obvious growth inhibition on Bel-7402 cells. The major mechanisms of inhibition included cell apoptosis and cytotoxic effects. Wogonin-induced cell death showed characteristics of apoptosis including DNA fragmentation, chromatin condensation, appearance of apoptotic bodies, and an increase in hypodiploid cells. However, the percentage of necrosis cells also increased with the increase of wogonin concentration. Furthermore, treatment with wogonin caused changes of reactive oxygen species (ROS) and mitochondrial membrane potentials (DeltaPsim, MMP), the decrease of the ratio of reduced and oxidized glutathione (GSH/GSSG), cytochrome c release and activation of caspase-9.


Macrophage polarization plays roles in bone formation instructed by calcium phosphate ceramics.
To investigate the roles of macrophages in material-instructed bone formation, two calcium phosphate (TCP) ceramics with the same chemistry but various scales of surface topography were employed in this study. After being implanted subcutaneously in FVB mice for 8 weeks, TCPs (TCP ceramics with submicron surface topography) gave rise to bone formation, while TCPb (TCP ceramics with micron surface topography) did not, showing the crucial role of surface topography scale in material-instructed bone formation. Depletion of macrophages with liposomal clodronate (LipClod) blocked such bone formation instructed by TCPs, confirming the role of macrophages in material-instructed bone formation. Macrophage cells (i.e. RAW 264.7 cells) cultured on TCPs in vitro polarized to tissue repair macrophages as evidenced by gene expression and cytokine production, while polarizing to pro-inflammatory macrophages on TCPb. Submicron surface topography of TCP ceramics directed macrophage polarization via PI3K/AKT pathways with the synergistic regulation of integrin β1. Finally, the tissue repair macrophage polarization on TCPs resulted in osteogenic differentiation of mesenchymal stem cells in vitro. At early implantation in FVB mice, TCPs recruited more macrophages which polarized towards tissue repair macrophages with time. The present data demonstrate the important roles of macrophage polarization in bone formation instructed by calcium phosphate ceramics.


A Multiplexed Serologic Test for Diagnosis of Lyme Disease for Point-of-Care Use.
Single multiplexed assays could replace the standard 2-tiered (STT) algorithm recommended for the laboratory diagnosis of Lyme disease if they perform with a specificity and a sensitivity superior or equal to those of the STT algorithm. We used human serum rigorously characterized to be sera from patients with acute- and convalescent-phase early Lyme disease, Lyme arthritis, and posttreatment Lyme disease syndrome, as well as the necessary controls (n = 241 samples), to select the best of 12 Borrelia burgdorferi proteins to improve our microfluidic assay (mChip-Ld). We then evaluated its serodiagnostic performance in comparison to that of a first-tier enzyme immunoassay and the STT algorithm. We observed that more antigens became positive as Lyme disease progressed from early to late stages. We selected three antigens (3Ag) to include in the mChip-Ld: VlsE and a proprietary synthetic 33-mer peptide (PepVF) to capture sensitivity in all disease stages and OspC for early Lyme disease. With the specificity set at 95%, the sensitivity of the mChip-Ld with 3Ag ranged from 80% (95% confidence interval [CI], 56% to 94%) and 85% (95% CI, 74% to 96%) for two panels of serum from patients with early Lyme disease and was 100% (95% CI, 83% to 100%) for serum from patients with Lyme arthritis; the STT algorithm detected early Lyme disease in the same two panels of serum from patients with early Lyme disease with a sensitivity of 48.5% and 75% and Lyme arthritis in serum from patients with Lyme arthritis with a sensitivity of 100%, and the specificity was 97.5% to 100%. The mChip-Ld platform outperformed the STT algorithm according to sensitivity. These results open the door for the development of a single, rapid, multiplexed diagnostic test for point-of-care use that can be designed to identify the Lyme disease stage.


Micro-fluidic chip platform for the characterization of breast cancer cells using aptamer-assisted immunohistochemistry.
We demonstrated a new QIHC (Quantitative Immunohistochemistry) microfluidic PDMS (Polydimethylsiloxane) platform by the introduction of the aptamer specific to the Fc region of the IgG antibody as a reporting probe. The aptamer was designed and synthesized. Various breast cancer cell lines were prepared as paraffin block slides, which were covered by a microfluidic PDMS platform to form a micro-reaction chamber. Primary antibodies specific to marker proteins (HER2, ER, PR, and ki-67) for breast cancer characterization were loaded in the micro-fluidic chip prior to the introduction of the aptamer. A master mixture of QNASBA (Quantitative Nucleic Acid Sequence Based Amplification) was used to quantify marker proteins by real time amplification of the aptamers. The quantitative results of aptamer amplification were linearly proportional to the concentrations of 4 different primary antibodies. The characterization results of the aptamer-assisted IHC using the microfluidic platform were well-correlated with those of conventional IHC for breast cancer cell lines (SK-BR-3, MCF-7, MDA-MB-231). Objective quantitative evaluations were carried out and compared with conventional results for real clinical samples.


Platelet-Rich Plasma Modulates Actions on Articular Cartilage Lubrication and Regeneration.
Osteoarthritis (OA) is characterized by articular cartilage degeneration in the joints and results in pain, swelling, stiffness, muscle atrophy, and attendant functional disability. The progression of OA cannot be prevented by drugs. Current treatments for OA focus on symptomatic relief of pain. Recently, platelet-rich plasma (PRP) injection has attracted much attention for modulating articular cartilage and synovial membrane in OA. Despite the increasing interest in PRP for the treatment of OA, the precise mechanisms underlying the actions of PRP on OA remain unclear. We have reviewed the known actions of PRP in the joint. The in vitro and in vivo evidence is reviewed concerning the potential of the modulation of PRP on normal articular cartilage and OA progression. PRP modulates the repair and regeneration of damaged articular cartilage in the joints and delays the degeneration of cartilage by stimulation of mesenchymal stem cell migration, proliferation, and differentiation into articular chondrocytes. In addition to the symptomatic relief, PRP is a biological response modifier of inflammatory nuclear factor-κB signaling pathway and PRP reduces the pain by decreasing inflammation and angiogenesis of the synovial membrane where pain receptors are localized. PRP has the therapeutic potential not only to promote tissue regeneration, but also to contribute to articular cartilage lubrication by decreasing the friction coefficient and minimizing wear. Although further refinements and improvements are needed in standardized PRP preparations, PRP may modulate regeneration of articular cartilage and retards the progression of OA by stimulating cell migration, proliferation, differentiation of progenitor/stem cells, joint homeostasis, and joint lubrication.


Implementation of an ultra-short-stay program after breast cancer surgery in four hospitals: perceived barriers and facilitators.
The objective of this study was to identify barriers and facilitators that professionals see when implementing a program incorporating ultra-short hospital admission in the treatment of breast cancer. Such an intervention is an essential step when designing a strategy for implementation of a care program that is different from established daily routines. In a prospective quasi-experimental study qualitative data were collected from four hospitals in the Netherlands between January 2005 and July 2006. Potential barriers and facilitators for successful implementation were extracted from detailed notes of all contacts between the researchers and each participating hospital. Subsequently, these items were categorized according to themes. Over 40 items were identified. Most barriers concerned organizational and program-related aspects, whereas the most common facilitators addressed organizational issues. Six of the 29 study recommendations were perceived as impeding or facilitating. Thirty of the 40 barriers were mentioned in one hospital only. Several key factors were found that determine the success of implementation of an ultrashort-stay program. Provision of care in the home setting should be assured. Policy makers and insurance companies should acknowledge that multidisciplinary care teams and teams integrating primary and secondary care fulfill important roles in delivering continuity of care. Specific strategies should be set out to convince everybody in the organization about the new ideas, particularly the minority of people who do not agree with the plans. A set of barriers and facilitators for implementation of the program was described that may be used by any professional preparing to perform breast cancer surgery in an ultrashort-stay facility. The systematic approach that led to this set may be used by any healthcare professional concerned with implementation and consolidation of innovative programs in healthcare in order to enhance the effectiveness of the chosen strategy.


Sacroiliac joint pain after multiple-segment lumbar fusion: a long-term observational study-Non-fused sacrum vs. fused sacrum.
Sacroiliac joint pain (SIJP) after lumbar fusion surgery has recently gained attention as a source of low back pain after lumbar fusion. There are two risk factors for postoperative SIJP, i.e., fusion involving the sacrum and multiple-segment fusion. In this study, we examined whether SIJP could occur more frequently in patients with two risk factors (multiple-segment fusion to sacrum). Further, we examined SIJP after multiple-segment (≥3) lumbar fusion, focusing on the difference between floating fusion (non-fused sacrum) and fixed fusion (fused sacrum). Ninety-one patients who underwent multiple-segment lumbar fusion were included. Patients without preoperative clinical SIJP were considered. Of these, 17 developed new-onset SIJP. We investigated postoperative SIJP development, duration from surgery to SIJP onset, and postoperative treatment outcomes of SIJP patients using Japanese Orthopaedic Association (JOA) scores. We compared the findings between floating fusion group and fixed fusion group. The incidence of SIJP was significantly higher with fixed fusion (32.1%) than with floating fusion (12.7%). The mean time of onset of sacroiliac joint pain was at 8.63 (2-13) months after surgery in the floating fusion group and 3.78 (1-10) months after surgery in the fixed fusion group, indicating that incidence occurred significantly earlier in the fixed fusion group. Our treatment outcome indicated that the mean JOA score significantly improved in the floating fusion group from 5.13 at the time of onset to 9.50 at the time of final follow-up; however, in the fixed fusion group, it improved from 5.78 at the time of onset to 7.33 at the time of final follow-up, indicating no significant improvement. In multiple-segment lumbar fusion, fixed fusion (fused sacrum) has a very high risk of SIJP. In addition, the onset of SIJP in such cases may occur earlier. This aspect deserves consideration, given the difficulty of pain treatment.


Severe hypertension after liver transplantation in alpha 1 antitrypsin deficiency.
Five children with alpha 1 antitrypsin deficiency and terminal liver disease received liver grafts; all five became hypertensive and four developed hypertensive encephalopathy. There was evidence of renal disease preoperatively and renal biopsy specimens showed variable glomerulonephritic histology with IgA nephropathy in one, mesangial-proliferative changes in two, and mesangio-capillary glomerulonephritis type I in two. Four hypertensive episodes were preceded by a fall in creatinine clearance. The association of glomerulonephritis with alpha 1 antitrypsin deficiency in children is more common than has been recognised. Affected patients are prone to severe hypertension of probable renal origin after liver transplantation and the renal lesion may affect long term prognosis.


The modern cardiovascular care unit: the cardiologist managing multiorgan dysfunction.
Despite many advances in the management of critically ill patients, cardiogenic shock remains a challenge because it is associated with high mortality. Even if there is no universally accepted definition of cardiogenic shock, end-perfusion organ dysfunction is an obligatory and major criterion of its definition.Organ dysfunction is an indicator that cardiogenic shock is already at an advanced stage and is undergoing a rapid self-aggravating evolution. The aim of the review is to highlight the importance to diagnose and to manage the organ dysfunction occurring in the cardiogenic shock patients by providing the best literature published this year. The first step is to diagnose the organ dysfunction and to assess their severity. Echo has an important and increasing place regarding the assessment of end-organ impairment whereas no new biomarker popped up. In this review, we aimed to highlight for intensivists and cardiologists managing cardiogenic shock, the recent advances in the care of end-organ dysfunctions associated with cardiogenic shock. The management of organ dysfunction is based on the improvement of the cardiac function by etiologic therapy, inotropes and assist devices but will often necessitate organ supports in hospitals with the right level of equipment and multidisciplinary expertise.


RAGE Plays a Role in LPS-Induced NF-κB Activation and Endothelial Hyperpermeability.
Endothelial functional dysregulation and barrier disruption contribute to the initiation and development of sepsis. The receptor for advanced glycation end products (RAGE) has been demonstrated to be involved in the pathogenesis of sepsis. The present study aimed to investigate the role of RAGE in lipopolysaccharide (LPS)-induced nuclear factor-κB (NF-κB) activation in endothelial cells and the consequent endothelial hyperpermeability. LPS-induced upregulation of RAGE protein expression in human umbilical vein endothelial cells (HUVECs) was detected by western blotting. Activation of NF-κB was revealed using western blotting and immunofluorescent staining. LPS-elicited endothelial hyperpermeability was explored by transendothelial electrical resistance (TER) assay and endothelial monolayer permeability assay. The blocking antibody specific to RAGE was used to confirm the role of RAGE in LPS-mediated NF-κB activation and endothelial barrier disruption. We found that LPS upregulated the protein expression of RAGE in a dose- and time-dependent manner in HUVECs. Moreover, LPS triggered a significant phosphorylation and degradation of IκBα, as well as NF-κB p65 nuclear translocation. Moreover, we observed a significant increase in endothelial permeability after LPS treatment. However, the RAGE blocking antibody attenuated LPS-evoked NF-κB activation and endothelial hyperpermeability. Our results suggest that RAGE plays an important role in LPS-induced NF-κB activation and endothelial barrier dysfunction.


Theoretical study of resonance formation in microhydrated molecules. I. Pyridine-(H2O)n, n = 1,2,3,5.
We present R-matrix calculations for electron scattering from microhydrated pyridine. We studied the pyridine-H2O cluster at static-exchange (SE), SE + polarization, and close-coupling levels, and pyridine-(H2O)n n = 2, 3, and 5 at SE level only in order to investigate the effect of hydrogen bonding on the resonances of pyridine. We analyse the results in terms of direct and indirect effects. We observe that the total (direct plus indirect) effect of microhydration leads to the stabilization of all resonances studied, both shape and core-excited. The size of the shift is different for different resonances and seems to be linked to the dipole moment of the cluster.


[Technical aspects and relevance of energy expenditure and physical activity assessment in clinical research for cystic fibrosis patients].
Cystic fibrosis (CF) is characterized by deteriorating lung function and mal-digestion, which result in growth failure and/or under-nutrition. Several factors, alone or combined, contribute to malnutrition in CF: poor energy intake, elevation of energy loss as a result of malabsorption, increasing resting energy expenditure due to genetic mutation and/or pulmonary exacerbation. Several techniques have been used to assess energy expenditure and physical activity in order to better understand mechanisms of malnutrition in CF and follow therapeutic interventions. Indirect calorimetry (IC) studies have shown that resting energy expenditure (REE) was 10-22% higher than predictive values. This increase could be attributed to chronic inflammation as a result of Pseudomonas aeruginosa (PA) infection. Indeed, intravenous antibiotic therapy decreases REE. Doubly labelled water technique and heart rate monitoring calibrated against IC techniques shows that total energy expenditure (TEE) was not different than in healthy children. Physical activity level assessed by the ratio TEE-REE is also not different between CF of healthy children. Recently, new accelerometry technics, easier to use and less invasive have been successfully used in order to assess physical activity level in CF. Precise and ambulatory assessment of energy expenditure and physical activity permit to check and adapt dietary allowances in CF. These techniques could be simultaneously used and be helpful to assess efficacy of intervention studies.


Reframing gene essentiality in terms of adaptive flexibility.
Essentiality assays are important tools commonly utilized for the discovery of gene functions. Growth/no growth screens of single gene knockout strain collections are also often utilized to test the predictive power of genome-scale models. False positive predictions occur when computational analysis predicts a gene to be non-essential, however experimental screens deem the gene to be essential. One explanation for this inconsistency is that the model contains the wrong information, possibly an incorrectly annotated alternative pathway or isozyme reaction. Inconsistencies could also be attributed to experimental limitations, such as growth tests with arbitrary time cut-offs. The focus of this study was to resolve such inconsistencies to better understand isozyme activities and gene essentiality. In this study, we explored the definition of conditional essentiality from a phenotypic and genomic perspective. Gene-deletion strains associated with false positive predictions of gene essentiality on defined minimal medium for Escherichia coli were targeted for extended growth tests followed by population sequencing and transcriptome analysis. Of the twenty false positive strains available and confirmed from the Keio single gene knock-out collection, 11 strains were shown to grow with longer incubation periods making these actual true positives. These strains grew reproducibly with a diverse range of growth phenotypes. The lag phase observed for these strains ranged from less than one day to more than 7 days. It was found that 9 out of 11 of the false positive strains that grew acquired mutations in at least one replicate experiment and the types of mutations ranged from SNPs and small indels associated with regulatory or metabolic elements to large regions of genome duplication. Comparison of the detected adaptive mutations, modeling predictions of alternate pathways and isozymes, and transcriptome analysis of KO strains suggested agreement for the observed growth phenotype for 6 out of the 9 cases where mutations were observed. Longer-term growth experiments followed by whole genome sequencing and transcriptome analysis can provide a better understanding of conditional gene essentiality and mechanisms of adaptation to such perturbations. Compensatory mutations are largely reproducible mechanisms and are in agreement with genome-scale modeling predictions to loss of function gene deletion events.


Factors Influencing Nutritional Adequacy among Rural Households in Nigeria: How Does Dietary Diversity Stand among Influencers?
This study examined the influence of food consumption diversity on adequate intakes of food calories, proteins and micronutrients among households in rural Nigeria within the framework of panel data econometrics using a nationally representative data. We found that substantial proportion of households suffered deficiency of calories, proteins and certain micronutrients; with higher percentage of sufferer households occurring in the post-planting season. The different measures of dietary diversity (constructed and used for analysis) consistently indicate significant and positive influence of dietary diversity on the likelihood of adequate consumption of food nutrients. While higher level of income, education and non-farm enterprise engagement may strongly stimulate adequate nutrient intakes, increases in the number of adolescents would substantially diminish it. Although our findings call for renewed attention on diet diverseness, we stress the complementary/synergistic roles of education and rural income improvement, especially through non-farm enterprise diversification in tackling multiple nutritional deficiencies in rural Nigeria.


Incidence of renal Fanconi syndrome in patients taking antiretroviral therapy including tenofovir disoproxil fumarate.
The objective of this study was to determine the incidence and predictors of Fanconi Syndrome (FS) in a cohort of patients taking tenofovir disoproxil fumarate (TDF). Clinical records and laboratory investigations from patients receiving TDF between 2002 and 2016 were extracted. FS was defined as normoglycaemic glycosuria and proteinuria and at least one other marker of renal dysfunction. Regression analysis was performed with time to development of FS and the following covariates: ritonavir co-administration, age, gender, co-morbidities (hypertension, hyperlipidaemia, diabetes, viral hepatitis), CD4 cell count nadir and baseline eGFR. One thousand and forty-four patients received TDF without ritonavir and 398 patients with ritonavir. Thirteen cases of FS were identified with a mean duration of exposure of 55 months. The incidence of FS was 1.09/1000PY (0.54-1.63) of TDF exposure (without ritonavir) and 5.50/1000PY (3.66-7.33) of TDF-ritonavir co-administration (p=0.0057). The adjusted hazards ratio for ritonavir co-administration was 4.71 (1.37-16.14, p=0.014). Known risk factors for chronic kidney disease were not associated with development of FS. Ritonavir co-administration, but not other factors, is associated with a greater risk of FS. FS developed late. Known risk factors for chronic kidney disease and length of treatment are not useful for identifying patients most at risk of developing FS in patients taking TDF.


Walking and bicycling: an evaluation of environmental audit instruments.
This paper reviews existing environmental audit instruments used to capture the walkability and bikability of environments. The review inventories and evaluates individual measures of environmental factors used in these instruments. It synthesizes the current state of knowledge in quantifying the built environment. The paper provides health promotion professionals an understanding of the essential aspects of environments influencing walking and bicycling for both recreational and transportation purposes. It serves as a basis to develop valid and efficient tools to create activity-friendly communities. Keyword searches identified journal articles from the computer-based Academic Citation Databases, including the National Transportation Library, the Web of Science Citation Database, and MEDLINE. Governmental publications and conference proceedings were also searched. All instruments to audit physical environments have been included in this review, considering both recreation- and transportation related walking and bicycling. Excluded are general methods devised to estimate walking and cycling trips, those used in empirical studies on land use and transportation, and research on walking inside buildings. Data have been extracted from each instrument using a template of key items developed for this review. The data were examined for quality assurance among three experienced researchers. A behavioral model of the built environment guides the synthesis according to three components: the origin and destination of the walk or bike trip, the characteristics of the road traveled, and the characteristics of the areas surrounding the trip's origin and destination. These components, combined with the characteristics of the instruments themselves, lead to a classification of the instruments into the four categories of inventory, route quality assessment, area quality assessment, and approaches to estimating latent demand for walking and bicycling. Furthermore, individual variables used in each instrument to measure the environment are grouped into four classes: spatiophysical, spatiobehavioral, spatiopsychosocial, and policy-based. Individually, existing instruments rely on selective classes of variables and therefore assess only parts of built environments that affect walking and bicycling. Most of the instruments and individual measures have not been rigorously tested because of a lack of available data on walking and bicycling and because of limited research budgets. Future instrument development will depend on the acquisition of empirical data on walking and bicycling, on inclusion of all three components of the behavioral model, and on consideration of all classes of variables identified.


Royal sun medicinal mushroom, Agaricus brasiliensis Ka21 (higher Basidiomycetes), as a functional food in humans.
The Royal Sun medicinal mushroom, Agaricus brasiliensis, is used as a natural health product. In Japan, however, the quality control of some of these mushroom products has been viewed as a safety problem. Focusing on the quality control of A. brasiliensis KA21, we have performed several safety studies. To date, we have established evidence that this mushroom can be used safely as an immunostimulant and to mediate biochemical parameters associated with obesity or diabetes. Furthermore, to improve the manufacturing process of this mushroom, we have studied the relationship between its pharmaceutical actions and the conditions of its cultivation and thermal management. The purpose of this review is to report the findings of basic and clinical studies of the fruit body of A. brasiliensis KA21.


Time-resolved fluorescence of the single tryptophan of Bacillus stearothermophilus phosphofructokinase.
The fluorescence of the single tryptophan in Bacillus stearothermophilus phosphofructokinase was characterized by steady-state and time-resolved techniques. The enzyme is a tetramer of identical subunits, which undergo a concerted allosteric transition. Time-resolved emission spectral data were fitted to discrete and distributed lifetime models. The fluorescence decay is a double exponential with lifetimes of 1.6 and 4.4 ns and relative amplitudes of 40 and 60%. The emission spectra of both components are identical with maxima at 327 nm. The quantum yield is 0.31 +/- 0.01. The shorter lifetime is independent of temperature; the longer lifetime has weak temperature dependence with activation energy of 1 kcal/mol. The fluorescence intensity and decay are the same in H2O and D2O solutions, indicating that the indole ring is not accessible to bulk aqueous solution. The fluorescence is not quenched significantly by iodide, but it is quenched by acrylamide with bimolecular rate constant of 5 x 10(8) M-1 s-1. Static and dynamic light scattering measurements show that the enzyme is a tetramer in solution with hydrodynamic radius of 40 A. Steady-state and time-resolved fluorescence anisotropies indicate that the tryptophan is immobile. The allosteric transition has little effect on the fluorescence properties. The fluorescence results are related to the x-ray structure.


Analgesic and tolerance studies with AP-237, a new analgesic.
1-n-Butyryl-4-cinnamylpiperazine hydrochloride, AP-237, is a new analgesic that has been reported in both Japan and the United States to have good analgesic activity in animals and is currently undergoing clinical trials. A detailed study of the analgesic and side effect profile of this compound is presented. AP-237 was an effective analgesic in a variety of species by several different routes of administration. Results of studies in an inflamed paw pressure and tail flick test in rats, hot plate and writhing test in mice, tooth pulp test in rabbits and intra-arterial bradykinin test in dogs are reported. AP-237 had a low physical dependence liability in rodents as measured by a mouse jumping test and a lack of withdrawal syndrom. Tolerance and cross tolerance results are reported. AP-237 is compared to morphine and pentazocine in most tests.


Taxometric analyses of borderline personality features in a large-scale male and female offender sample.
Few studies to date have examined the extent to which borderline personality features are best construed as representing an underlying dimension or a discrete class or taxon. The authors conducted taxometric analyses using a large-scale sample of male (n=787) and female (n=368) prison inmates who had completed the Personality Assessment Inventory (L. C. Morey, 2007). Analyses based on the 4 subscales of the Borderline Features Scale offered compelling support for a dimensional structure in both the full sample and the female subsample--even after controlling for the potentially confounding effects of negative response distortion. Theoretical and pragmatic implications of these findings are reviewed.


A new mechanism for void-cascade interaction from nondestructive depth-resolved atomic-scale measurements of ion irradiation-induced defects in Fe.
The nondestructive investigation of single vacancies and vacancy clusters in ion-irradiated samples requires a depth-resolved probe with atomic sensitivity to defects. The recent development of short-pulsed positron beams provides such a probe. Here, we combine depth-resolved Doppler broadening and positron annihilation lifetime spectroscopies to identify vacancy clusters in ion-irradiated Fe and measure their density as a function of depth. Despite large concentrations of dislocations and voids in the pristine samples, positron annihilation measurements uncovered the structure of vacancy clusters and the change in their size and density with irradiation dose. When combined with transmission electron microscopy measurements, the study demonstrates an association between the increase in the density of small vacancy clusters with irradiation and a remarkable reduction in the size of large voids. This, previously unknown, mechanism for the interaction of cascade damage with voids in ion-irradiated materials is a consequence of the high porosity of the initial microstructure.


The Immunosuppressant Macrolide Tacrolimus Activates Cold-Sensing TRPM8 Channels.
TRPM8 is a polymodal, nonselective cation channel activated by cold temperature and cooling agents that plays a critical role in the detection of environmental cold. We found that TRPM8 is a pharmacological target of tacrolimus (FK506), a macrolide immunosuppressant with several clinical uses, including the treatment of organ rejection following transplants, treatment of atopic dermatitis, and dry eye disease. Tacrolimus is an inhibitor of the phosphatase calcineurin, an action shared with cyclosporine. Tacrolimus activates TRPM8 channels in different species, including humans, and sensitizes their response to cold temperature by inducing a leftward shift in the voltage-dependent activation curve. The effects of tacrolimus on purified TRPM8 in lipid bilayers demonstrates conclusively that it has a direct gating effect. Moreover, the lack of effect of cyclosporine rules out the canonical signaling pathway involving the phosphatase calcineurin. Menthol (TRPM8-Y745H)- and icilin (TRPM8-N799A)-insensitive mutants were also activated by tacrolimus, suggesting a different binding site. In cultured mouse DRG neurons, tacrolimus evokes an increase in intracellular calcium almost exclusively in cold-sensitive neurons, and these responses were drastically blunted in Trpm8 KO mice or after the application of TRPM8 antagonists. Cutaneous and corneal cold thermoreceptor endings are also activated by tacrolimus, and tacrolimus solutions trigger blinking and cold-evoked behaviors. Together, our results identify TRPM8 channels in sensory neurons as molecular targets of the immunosuppressant tacrolimus. The actions of tacrolimus on TRPM8 resemble those of menthol but likely involve interactions with other channel residues.SIGNIFICANCE STATEMENT TRPM8 is a polymodal TRP channel involved in cold temperature sensing, thermoregulation, and cold pain. TRPM8 is also involved in the pathophysiology of dry eye disease, and TRPM8 activation has antiallodynic and antipruritic effects, making it a prime therapeutic target in several cutaneous and neural diseases. We report the direct agonist effect of tacrolimus, a potent natural immunosuppressant with multiple clinical applications, on TRPM8 activity. This interaction represents a novel neuroimmune interface. The identification of a clinically approved drug with agonist activity on TRPM8 channels could be used experimentally to probe the function of TRPM8 in humans. Our findings may explain some of the sensory and anti-inflammatory effects described for this drug in the skin and the eye surface.


β-lapachone: A Promising Anticancer Agent with a Unique NQO1 Specific Apoptosis in Pancreatic Cancer.
Cancer, one of the major health problems all over the world, requires more competent drugs for clinical use. One recent possible chemotherapeutic drug under research is β-lapachone. β- lapachone (1,2-naphthoquinone) has promising activity against those tumors showing raised levels of Nicotinamide di-phosphate Quinone Oxidoreductases-1 (NQO1). NQO1 is found to be up-regulated in pancreatic tumor cells, and thus β-lapachone could generate cytotoxicity in various cancers like pancreatic tumors. β-lapachone harborage independent growth and clonogenic cell survival in agar. The cell-killing effects of β-lapachone can be stopped by using dicumarol, an inhibitor of NAD(P)H Quinone Oxidoreductases-1. In previously established pancreatic cancer xenografts in mice, β- lapachone inhibited the tumor growth when given orally rather than when combined with cyclodextrin to improve its bioavailability.


The Electronic Bee Spy: Eavesdropping on Honeybee Communication via Electrostatic Field Recordings.
As a canary in a coalmine warns of dwindling breathable air, the honeybee can indicate the health of an ecosystem. Honeybees are the most important pollinators of fruit-bearing flowers, and share similar ecological niches with many other pollinators; therefore, the health of a honeybee colony can reflect the conditions of a whole ecosystem. The health of a colony may be mirrored in social signals that bees exchange during their sophisticated body movements such as the waggle dance. To observe these changes, we developed an automatic system that records and quantifies social signals under normal beekeeping conditions. Here, we describe the system and report representative cases of normal social behavior in honeybees. Our approach utilizes the fact that honeybee bodies are electrically charged by friction during flight and inside the colony, and thus they emanate characteristic electrostatic fields when they move their bodies. These signals, together with physical measurements inside and outside the colony (temperature, humidity, weight of the hive, and activity at the hive entrance) will allow quantification of normal and detrimental conditions of the whole colony. The information provided instructs how to setup the recording device, how to install it in a normal bee colony, and how to interpret its data.


The place of osmophobia and taste abnormalities in migraine classification: a tertiary care study of 1237 patients.
This study evaluates osmophobia and taste abnormalities in relationship to sensitivity and specificity in the classification of migraine. Consecutive International Headache Society (IHS) classified patients (n = 1237) were evaluated. Symptoms were graded from 0 to 3. Osmophobia and taste abnormalities were tested for sensitivity and specificity in migraine diagnosis. The patients were 85.4% female and their mean age was 38.1 years. Of 673 patients 24.7% complained of osmophobia, and 24.6% of 505 complained of taste abnormalities. In the absence of nausea and vomiting the combinations of two symptoms gave the following sensitivity and specificity percentages, respectively: photophobia and phonophobia, 10.6 and 84.9; photophobia and osmophobia, 1.1 and 99.0; phonophobia and osmophobia, 1.1 and 98.6; photophobia and taste abnormality, 9.6 and 99.0; phonophobia and taste abnormality, 9.6 and 98.8; and osmophobia and taste abnormality, 4.2 and 99.4. Osmophobia and taste abnormalities were demonstrated to be very specific in diagnosing migraine IHS 1.1-1.6, but very insensitive.


[Clinical aspects of trimetazidine (preductal) administration as an antianginal drug].
To study antianginal effectiveness of trimetazidine (TMS) in monotherapy and combined antianginal therapy; to design approaches to prediction of its effect. An open cross-over randomised trial enrolled 29 anginal patients. They were given a single TMS dose (60 mg) and its effect was compared to that of placebo, propranolol, verapamil, isosorbide dinitrate, nifedipine). The effect was estimated by an increment of bicycle exercise (BE) duration necessary to depress ST segment by 1 mm. 10 patients retained signs of myocardial ischemia at BE though they were treated with specially selected antianginal drugs. In them, we assessed additional benefit of adjuvant TMS. The drugs were tried under acute test and one-month treatment. In acute test a significant antianginal effects of TMS were registered against the placebo. Differences in efficiency of the antianginal drugs were insignificant. The addition of TMS to the selected drugs gave rise to the trend to prolongation of time to ST segment depression by 1 mm. We found a strong correlation between TMS efficiency in acute pharmacological test and in long-term administration. This enables using BE for prediction of long-term effect of the drug. TMS has a significant antianginal effect. It is especially beneficial in cases of insufficient effectiveness of conventional antianginal drugs or when their use is problematic because of marked disturbances of hemodynamics.


Regulation of intracellular Mg2+ by superoxide in amnion cells.
Changes of intracellular free Mg2+ concentration ([Mg2+]i) in human amnion cells induced by superoxide anion were determined using a highly Mg(2+)-sensitive fluorescent dye Mg(2+)-fura2 or Mg(2+)-indol. Superoxide anion, produced by addition of xanthine oxidase to hypoxanthine, induced decrease of [Mg2+]i. The decrease was significantly inhibited by an anion channel blocker, 4,4'diisothiocyano-2,2' disulfonic acid stilbene (DIDS). Superoxide dismutase (SOD), injected into cells by cell fusion, also inhibited the change of [Mg2+]i, but catalase did not. Superoxide anion induced prompt increase of intracellular pH (pHi) as well as decrease of [Mg2+]i and subsequently activated the increase of intracellular free Ca2+ ([Ca2+]i) and the release of arachidonate. In contrast to superoxide anion, NH4Cl which induces increase of pHi in amnion cells increased [Mg2+]i. The elevation of basal level of [Mg2+]i by Mg(2+)-ionophore inhibited the change of [Ca2+]i and the release of arachidonate induced by superoxide anion. These results suggest that superoxide anion, transported through anion channels into cells, decreases [Mg2+]i directly, not due to a pH-effect and that the decrease of [Mg2+]i may regulate biological functions of the cells via increase of [Ca2+]i.


Dermoscopy in Leprosy: A Clinical and Histopathological Correlation Study.
Leprosy, an insidious infectious granulomatous disease, is diagnosed traditionally through clinical examination coupled with skin smears and histopathology. It has myriad clinical presentations that pose diagnostic challenges. Lately, dermoscopy has emerged as a rapid, noninvasive diagnostic modality for many dermatoses. We evaluated the dermoscopic findings of various manifestations of leprosy and correlated them with clinical and histopathological features. This prospective, cross-sectional study was conducted in our skin outpatient department for a period of 1 year. Patients newly diagnosed as having leprosy or those undergoing leprosy treatment for less than 6 months were included. The most representative lesion was dermoscopically evaluated and later biopsied. We included 73 patients in the study. Results indicated an obvious correlation between dermoscopic findings and histopathology. We noted orangish yellow and white structureless areas, steadily throughout the spectrum, depicting dermal granuloma. Additionally, we observed focal vascular structures such as branching, linear, and crown vessels that result from the pressure of granuloma pushing the dilated vessels upwards. The relative absence of skin appendages aided in differentiating leprosy from other granulomatous disorders. Novel findings of our study were the detection of a branch-like pattern of clofazimine-induced pigmentation on dermoscopy and orange globules on onychoscopy. Other unique findings included violaceous structureless areas, characteristic large telangiectatic vessels, follicular plugging, star-shaped silvery-white scaling, and white globules in type 1 reaction; white shiny steaks were observed in patients with borderline lepromatous leprosy, and central white dots and keratotic plugs were observed in patients with histoid leprosy. Dermoscopy, as a noninvasive modality, could aid in the quick diagnosis of leprosy and should be used as a handy tool to complement other investigative tools for this disease.


Effects of pharmacologic antagonists of epidermal growth factor receptor, PI3K and MEK signal kinases on NF-kappaB and AP-1 activation and IL-8 and VEGF expression in human head and neck squamous cell carcinoma lines.
We previously reported that expression of angiogenesis factors interleukin-8 (IL-8) and vascular endothelial growth factor (VEGF) is promoted by coactivation of transcription factors nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1) by interleukin-1alpha in human head and neck squamous cell carcinomas (HNSCC). However, expression of IL-1 receptor antagonist incompletely blocked reporter gene activity and cytokine expression, suggesting that other upstream signals may contribute to activation. Overexpression and autocrine activation of epidermal growth factor receptor (EGFR) is detected in 90% of HNSCC, and EGFR inhibitors have been reported to inhibit IL-8 and VEGF expression, but the intermediary signal pathways and transcription factors by which EGFR modulates proangiogenic factors is unknown. EGFR can activate the phosphotidylinositol-3 kinase (PI3K) and mitogen-activated/extracellular signal-regulated kinase (MEK) pathways, which can potentially modulate activation of NF-kappaB and AP-1, respectively. In our study, we examined the effect of EGF and antagonists of EGFR, PI3K and MEK on NF-kappaB and AP-1 activation and IL-8 and VEGF expression in HNSCC cell lines UM-SCC-9 and 11B in which EGFR is overexpressed and activated. Recombinant EGF induced EGFR phosphorylation, activation of NF-kappaB and AP-1 reporter genes and IL-8 and VEGF expression, indicating that EGFR can mediate coactivation of both transcription factors and cytokine genes in HNSCC. EGFR antagonist PD153035 and anti-EGFR antibody C225 completely inhibited EGF-induced reporter activity and cytokine expression, but only partially inhibited constitutive activity. MEK inhibitor U0126 preferentially blocked AP-1 activity and expression of both IL-8 and VEGF, while PI3K inhibitor LY-294002 or a dominant negative inhibitor-kappaB preferentially blocked NF-kappaB activation and expression of IL-8 but not VEGF. EGFR, PI3K and MEK antagonists inhibited growth of HNSCC. We conclude that antagonists of EGFR, PI3K and MEK signal pathways have inhibitory activity against EGFR-induced NF-kappaB and AP-1 activation, IL-8 and VEGF expression and growth by HNSCC. Published 2002 Wiley-Liss, Inc.


['Will I receive a liver transplant in time?'; chance of survival of patients on the liver transplant waiting list].
To calculate the chance of receiving a liver transplant for patients on the liver transplant waiting list in the Netherlands. Retrospective cohort research. Data of all patients in the Netherlands on the waiting list for liver transplantation, from the introduction of the model of end-stage liver disease score on 16th December 2006 through to 31st December 2013 were collected. Survival analysis was computed with competing risk analyses. A total of 851 patients were listed, of whom 236 patients with hepatocellular carcinoma, 147 patients with primary sclerosing cholangitis, 142 patients with post-alcoholic liver disease, 93 patients with metabolic liver disease, 78 with viral hepatitis and 155 patients listed for other indications. The median waiting time till transplantation was 196 days. The chance to be transplanted at two years from listing was 65% and the risk of death was 17%. Patients with metabolic liver disease had the highest chance of undergoing liver transplantation. Patients with viral hepatitis were at highest risk of death while on the list, as well as having the lowest chance of undergoing liver transplantation. Our study shows a 65% chance of getting transplanted in time after a median waiting time of 6 months in the Netherlands. Sadly, 1 in 6 patients die before liver transplantation can be performed, with the highest risk of death occurring in patients with viral hepatitis.


Residential proximity to abandoned uranium mines and serum inflammatory potential in chronically exposed Navajo communities.
Members of the Navajo Nation, who possess a high prevalence of cardiometabolic disease, reside near hundreds of local abandoned uranium mines (AUM), which contribute uranium, arsenic and other metals to the soil, water and air. We recently reported that hypertension is associated with mine waste exposures in this population. Inflammation is a major player in the development of numerous vascular ailments. Our previous work establishing that specific transcriptional responses of cultured endothelial cells treated with human serum can reveal relative circulating inflammatory potential in a manner responsive to pollutant exposures, providing a model to assess responses associated with exposure to these waste materials in this population. To investigate a potential link between exposures to AUM and serum inflammatory potential in affected communities, primary human coronary artery endothelial cells were treated for 4 h with serum provided by Navajo study participants (n=145). Endothelial transcriptional responses of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and chemokine ligand 2 (CCL2) were measured. These transcriptional responses were then linked to AUM exposure metrics, including surface area-weighted AUM proximity and estimated oral intake of metals. AUM proximity strongly predicted endothelial transcriptional responses to serum including CCL2, VCAM-1 and ICAM-1 (P<0.0001 for each), whereas annual water intakes of arsenic and uranium did not, even after controlling for all major effect modifiers. Inflammatory potential associated with proximity to AUMs, but not oral intake of specific metals, additionally suggests a role for inhalation exposure as a contributor to cardiovascular disease.


[Immunotherapy in childhood asthma].
It was accomplished a prospective study with 100 pediatric patients (60 children- and 40 girls) with the diagnoses of chronic rhinitis and bronchial asthma and reactivity allergic to three or more antigens environmental related to exacerbation of disease and whose symptoms were impeding a normal life. It was then included in a program of immunotherapy in compliance with the plan that has in the Centro Regional para la Prevención y el Tratamiento de las Enfermedades Alérgicas (CRPTEA) of the Hospital Universitario de la Universidad Autónoma de Nuevo León, as well as an education and rehabilitation program of the patient and family. There was a notable improvement, clinically demonstrated by the important decrease in the quantity of asthmatic crises and rapid answer to drugs, being integrated to a life of better quality, with good scholastic exploitation, suitable allowance to exercise and quiet sleep. It was concluded that, in well selected asthmatic patients, it is justified the immunotherapy, when it can be demonstrated specific sensitisation to allergens by tests cutaneous and that unfetter the symptoms and that they do not answer adequately to pharmacotherapy of first line.


MicroRNA expression profile in myocardial bridging patients.
Myocardial bridging (MB), a common benign coronary anomaly, may bring about some unwanted complications such as angina-like chest pain. The only way of MB detection is coronary arteriography or coronary computed tomographic angiography, which is costly and invasive. This study intended to profile a panel of circulating microRNAs (miRNAs) as potential biomarkers for the diagnosis of MB. Using TaqMan Low-Density Array followed by quantitative reverse transcriptase PCR (qRT-PCR) validation, we analyzed the expression of miRNAs in serum samples from 90 MB patients and 50 non-MB controls. The Low-Density Array data showed that 196 miRNAs were differentially expressed in MB patient sera in comparison with controls. After qRT-PCR validation and receiver operating characteristic (ROC) analysis, a list of five miRNAs (miR-29b, miR-151-3p, miR-126, miR-503-3p and miR-645) showed the ability to distinguish MB patients from controls. The area under curve (AUC) values range from 0.722-0.938. We have demonstrated that this panel of five serum miRNAs is expected to become potential non-invasive biomarkers for detection of MB.


Clinical features and oncologic outcomes in patients with rectal cancer and ulcerative colitis: a single-institution experience.
Patients with chronic ulcerative colitis are at increased risk of developing colorectal cancer. Limited data exists in ulcerative colitis patients with rectal cancer regarding clinical and oncologic outcomes, and the ideal operative approach. To describe our experience in the management of patients with rectal cancer in the setting of chronic ulcerative colitis and their outcomes. This study is a retrospective review of all patients with ulcerative colitis who underwent a colorectal operation between 1990 and 2009. This study was conducted at a tertiary care center. Adult patients with rectal adenocarcinoma undergoing a colorectal operation for chronic ulcerative colitis were included in this study. Patients with colonic malignancy, indeterminate colitis, and Crohn's disease were excluded. Clinical features and long-term oncologic outcomes are described. Forty-one patients were identified; their mean age was 53.9 years. Mean duration of ulcerative colitis was 22.5 years. Thirty-four patients (83%) were known to have cancer preoperatively; in 7 patients it was discovered on postoperative pathology. Eight of the tumors were in the proximal rectum, 19 in the mid rectum, and 13 in the distal rectum. The most common operation performed was total proctocolectomy with end ileostomy (n = 21), followed by IPAA. The majority of patients (n = 28, 68%) had stage I or II disease. Estimates of overall survival at 1 and 5 years were 83% and 62%, and, for disease-free survival, the estimates were 93% and 62%. Local and distant recurrence was seen in 5 and 9 patients. Eighty-nine percent of the observed recurrences were in patients with stage III and IV disease. Pouch failure occurred in 2 patients. This retrospective study was possibly underpowered, given the small sample sizes. In our cohort, rectal cancer in the setting of chronic ulcerative colitis was rare, often presented at an early stage, and was not always diagnosed preoperatively. The presence of early-stage rectal cancer should not be considered a contraindication for IPAA.


Vector description of higher-order modes in photonic crystal fibers
We extensively study the propagation features of higher-order modes in a photonic crystal fiber (PCF). Our analysis is based on a full-vector modal technique specially adapted to accurately describe light propagation in PCF's. Unlike conventional fibers, PCF's exhibit a somewhat unusual mechanism for the generation of higher-order modes. Accordingly, PCF's are characterized by the constancy of the number of modes below a wavelength threshold. An explicit verification of this property is given through a complete analysis of the dispersion relations of higher-order modes in terms of the structural parameters of this kind of fiber. The transverse irradiance distributions for some of these higher-order modes are also presented, showing an excellent agreement with recent experimental measurements. In the same way, the full-vector nature of our approach allows us to analyze the rich polarization structure of the PCF mode spectrum.


Molecular and Biochemical Insights Into Early Responses of Hemp to Cd and Zn Exposure and the Potential Effect of Si on Stress Response.
With the intensification of human activities, plants are more frequently exposed to heavy metals (HM). Zinc (Zn) and cadmium (Cd) are frequently and simultaneously found in contaminated soils, including agronomic soils contaminated by the atmospheric fallout near smelters. The fiber crop Cannabis sativa L. is a suitable alternative to food crops for crop cultivation on these soils. In this study, Cd (20 μM) and Zn (100 μM) were shown to induce comparable growth inhibition in C. sativa. To devise agricultural strategies aimed at improving crop yield, the effect of silicon (Si; 2 mM) on the stress tolerance of plants was considered. Targeted gene expression and proteomic analysis were performed on leaves and roots after 1 week of treatment. Both Cd- and Zn-stimulated genes involved in proline biosynthesis [pyrroline-5-carboxylate reductase (P5CR)] and phenylpropanoid pathway [phenylalanine ammonia-lyase (PAL)] but Cd also specifically increased the expression of PCS1-1 involved in phytochelatin (PC) synthesis. Si exposure influences the expression of numerous genes in a contrasting way in Cd- and Zn-exposed plants. At the leaf level, the accumulation of 122 proteins was affected by Cd, whereas 47 proteins were affected by Zn: only 16 proteins were affected by both Cd and Zn. The number of proteins affected due to Si exposure (27) alone was by far lower, and 12 were not modified by heavy metal treatment while no common protein seemed to be modified by both CdSi and ZnSi treatment. It is concluded that Cd and Zn had a clear different impact on plant metabolism and that Si confers a specific physiological status to stressed plants, with quite distinct impacts on hemp proteome depending on the considered heavy metal.


A Mobile Phone-Based Program to Promote Healthy Behaviors Among Adults With Prediabetes Who Declined Participation in Free Diabetes Prevention Programs: Mixed-Methods Pilot Randomized Controlled Trial.
Despite evidence that Diabetes Prevention Programs (DPPs) can delay or prevent progression to type 2 diabetes mellitus (T2DM), few individuals with prediabetes enroll in offered programs. This may be in part because many individuals with prediabetes have low levels of autonomous motivation (ie, motivation that arises from internal sources) to prevent T2DM. This study aims to examine the feasibility and acceptability of a mobile health (mHealth) intervention designed to increase autonomous motivation and healthy behaviors among adults with prediabetes who previously declined participation free DPPs. In addition, the study aims to examine changes in autonomous motivation among adults offered 2 versions of the mHealth program compared with an information-only control group. In this 12-week, parallel, 3-arm, mixed-methods pilot randomized controlled trial, participants were randomized to (1) a group that received information about prediabetes and strategies to prevent T2DM (control); (2) a group that received a mHealth app that aims to increase autonomous motivation among users (app-only); or (3) a group that received the app plus a physical activity tracker and wireless-enabled digital scale for self-monitoring (app-plus). Primary outcome measures included rates of intervention uptake (number of individuals enrolled/number of individuals assessed for eligibility), retention (number of 12-week survey completers/number of participants), and adherence (number of device-usage days). The secondary outcome measure was change in autonomous motivation (measured using the Treatment Self-Regulation Questionnaire), which was examined using difference-in-difference analysis. Furthermore, we conducted postintervention qualitative interviews with participants. Overall, 28% (69/244) of eligible individuals were randomized; of these, 80% (55/69) completed the 12-week survey. Retention rates were significantly higher among app-plus participants than participants in the other 2 study arms combined (P=.004, χ2). No significant differences were observed in adherence rates between app-only and app-plus participants (43 days vs 37 days; P=.34). Among all participants, mean autonomous motivation measures were relatively high at baseline (6.0 of 7.0 scale), with no statistically significant within- or between-group differences in follow-up scores. In qualitative interviews (n=15), participants identified reasons that they enjoyed using the app (eg, encouraged self-reflection), reasons that they did not enjoy using the app (eg, did not consider personal circumstances), and strategies to improve the intervention (eg, increased interpersonal contact). Among individuals with prediabetes who did not engage in free DPPs, this mHealth intervention was feasible and acceptable. Future work should (1) examine the effectiveness of a refined intervention on clinically relevant outcomes (eg, weight loss) among a larger population of DPP nonenrollees with low baseline autonomous motivation and (2) identify other factors associated with DPP nonenrollment, which may serve as additional potential targets for interventions. ClinicalTrials.gov NCT03025607; https://clinicaltrials.gov/ct2/show/NCT03025607 (Archived by WebCite at http://www.webcitation.org/73cvaSAie).


Characterization, phylogeny, alternative splicing and expression of Sox30 gene.
Members of the Sox gene family isolated from both vertebrates and invertebrates have been proved to participate in a wide variety of developmental processes, including sex determination and differentiation. Among these members, Sox30 had been considered to exist only in mammals since its discovery, and its exact function remains unclear. Sox30 cDNA was cloned from the Nile tilapia by RT-PCR and RACE. Screening of available genome and EST databases and phylogenetic analysis showed that Sox30 also exists in non-mammalian vertebrates and invertebrates, which was further supported by synteny analyses. Tissue expression in human, mouse and tilapia suggested that Sox30 was probably a gonad-specific gene, which was also supported by the fact that Sox30 EST sequences were obtained from gonads of the animal species. In addition, four alternatively spliced isoforms were isolated from tilapia gonad. Their temporal and spatial expression patterns during normal and sex reversed gonadal development were investigated by RT-PCR and in situ hybridization. Our data suggest that expressions of Sox30 isoforms are related to stage and phenotypic-sex, observed in the germ cells of male gonad and in somatic cells of the female gonad. Sox30 is not a gene only existed in mammals, but exists widely throughout the animal kingdom as supported by our bioinformatic, phylogenetic and syntenic analyses. It is very likely that Sox30 is expressed exclusively in gonads. Expression analyses revealed that Sox30 may be involved in female and male gonadal development at different stages by alternative splicing.


Non-convulsive status epilepticus associated with glutamic acid decarboxylase antibody.
Autoimmune encephalitis associated with glutamic acid decarboxylase antibodies (GAD-Ab) often presents with treatment-resistant partial seizures, as well as other central nervous system symptoms. In contrast to several other well-characterized autoantibodies, GAD-Ab has very rarely been associated with status epilepticus. We report a 63-year-old woman initially admitted with somnolence and psychiatric findings. The EEG findings, of generalized and rhythmical slow spike-wave activity over the posterior regions of both hemispheres, together with the clinical deterioration in responsiveness, led to the diagnosis of non-convulsive status epilepticus. Investigation of a broad panel of autoantibodies, revealed only increased serum GAD-Ab levels. Following methylprednisolone and intravenous immunoglobulin treatments, the patient's neurological symptoms improved, EEG findings disappeared and GAD-Ab levels significantly decreased. GAD-Ab should be added to the list of anti-neuronal antibodies associated with non-convulsive status epilepticus. Disappearance of clinical findings and seroreversion after immunotherapy suggest that GAD-Ab might be involved in seizure pathogenesis.


Tourette's disorder and related problems: a review and update.
Tourette's disorder is a complex, multifaceted condition of neurological origin with psychiatric symptomatology characterized by multiple motor and vocal tics. It begins during childhood and has a waxing and waning course. Coexisting obsessive-compulsive symptoms, attentional problems, and other behavioral features are common. Pathophysiology involves dysfunction in basal ganglia and related corticothalamic circuits. Many patients who present in clinical settings have mild to moderate symptoms and require only education, monitoring, and long-term follow-up. Those with more-severe symptoms require treatment, typically including both pharmacotherapy and nonmedication approaches. In addition to traditional neuroleptic treatment, a variety of agents such as clonidine, serotonin-reuptake inhibitors, and tricyclic antidepressants can be used. Further investigation is needed in a variety of areas, including longitudinal follow-up studies of children, adolescents, and adults with Tourette's disorder; systematic investigation of psychiatric comorbidity and the heterogeneity of the disorder; clarification of the phenomenological similarities and differences between Tourette's disorder and obsessive-compulsive disorder; and neuroimaging and neuropsychological studies of Tourette's disorder and related problems.


Measles control in Australia - threats, opportunities and future needs.
Control of measles was the focus of a national workshop held in 2015 in Sydney, Australia, bringing together stakeholders in disease control and immunisation to discuss maintaining Australia's measles elimination status in the context of regional and global measles control. The global epidemiology of measles was reviewed, including outbreaks in countries that have achieved elimination, such as the Disneyland outbreak in the United States and large outbreaks in Sydney, Australia. Transmission of measles between Australia and New Zealand occurs, but has not been a focus of control measures. Risk groups, the genetic and seroepidemiology of measles as well as surveillance, modelling and waning vaccine-induced immunity were reviewed. Gaps in policy, research and practice for maintaining measles elimination status in Australia were identified and recommendations were developed. Elimination of measles globally is challenging because of the infectiousness of measles and the need for 2-dose vaccine coverage rates in excess of 95% in all countries to achieve it. Until this occurs, international travel will continue to permit measles importation from endemic countries to countries that have achieved elimination. When measles cases are imported, failure to diagnose and isolate cases places the health system at risk of measles outbreaks. Vaccine funding models can result in gaps in vaccine coverage for adults and migrants. Australia introduced a whole-of-life immunisation register in 2016 and catch-up vaccination for at-risk communities, which will improve measles control. Research on diagnosis, immunology, case management and modelling of vaccination strategies are important to ensure continued control of measles.


A developmental staging table for Astyanax mexicanus surface fish and Pachón cavefish.
Every model species requires its own developmental table. Astyanax mexicanus, a teleost fish comprising both sighted river and blind cave populations, is becoming more and more important in the field of developmental and evolutionary biology. As such, a developmental staging table is increasingly necessary, particularly since comparative analysis of early developmental events is widely employed by researchers. We collected freshly spawned embryos from surface fish and Pachón cavefish populations. Embryos were imaged every 10-12 min during the first day of development, and less frequently in the following days. The results provide an illustrated comparison of selected developmental stages from one cell to hatching of these two populations. The two morphs show an essentially synchronous development regarding major events such as epiboly, neurulation, somitogenesis, heart beating, or hatching. We also present data on particular morphological characters appearing during larval development, such as eye size, yolk regression, swim bladder, and fin development. Some details about the development of F1 Pachón cave×surface hybrids are also given. Comparisons are made with Danio rerio (zebrafish) development.


Anaesthetic management of the morbidly obese.
The problems associated with obesity are brought into sharp focus at the time of a surgical operation. The anaesthetist has a major role to play in preventing serious complications and must be aware of the many potential hazards. Extra vigilance is required to guide these patients safely through the perioperative period.


Ser67Asp and His68Asp substitutions in candida parapsilosis carbonyl reductase alter the coenzyme specificity and enantioselectivity of ketone reduction.
A short-chain carbonyl reductase (SCR) from Candida parapsilosis catalyzes an anti-Prelog reduction of 2-hydroxyacetophenone to (S)-1-phenyl-1,2-ethanediol (PED) and exhibits coenzyme specificity for NADPH over NADH. By using site-directed mutagenesis, the mutants were designed with different combinations of Ser67Asp, His68Asp, and Pro69Asp substitutions inside or adjacent to the coenzyme binding pocket. All mutations caused a significant shift of enantioselectivity toward the (R)-configuration during 2-hydroxyacetophenone reduction. The S67D/H68D mutant produced (R)-PED with high optical purity and yield in the NADH-linked reaction. By kinetic analysis, the S67D/H68D mutant resulted in a nearly 10-fold increase and a 20-fold decrease in the k(cat)/K(m) value when NADH and NADPH were used as the cofactors, respectively, but maintaining a k(cat) value essentially the same with respect to wild-type SCR. The ratio of K(d) (dissociation constant) values between NADH and NADPH for the S67D/H68D mutant and SCR were 0.28 and 1.9 respectively, which indicates that the S67D/H68D mutant has a stronger preference for NADH and weaker binding for NADPH. Moreover, the S67D/H68D enzyme exhibited a secondary structure and melting temperature similar to the wild-type form. It was also found that NADH provided maximal protection against thermal and urea denaturation for S67D/H68D, in contrast to the effective protection by NADP(H) for the wild-type enzyme. Thus, the double point mutation S67D/H68D successfully converted the coenzyme specificity of SCR from NADP(H) to NAD(H) as well as the product enantioselectivity without disturbing enzyme stability. This work provides a protein engineering approach to modify the coenzyme specificity and enantioselectivity of ketone reduction for short-chain reductases.


Serum Wisteria floribunda agglutinin-positive Mac-2 binding protein in non-alcoholic fatty liver disease.
Wisteria floribunda agglutinin-positive Mac-2 binding protein (WFA+-M2BP) has been suggested to be useful for the assessment of disease severity in non-alcoholic fatty liver disease (NAFLD). Consecutive adult NAFLD patients who had a liver biopsy were included. Serum WFA+-M2BP level was measured using a lectin-antibody sandwich immunoassay using a chemiluminescence enzyme immunoassay machine (HISCL-5000, Sysmex, Kobe, Japan). The measured levels were indexed using the following equation: Cut-off index (COI) = ([WFA+-M2BP]sample-[WFA+-M2BP]NC) / ([WFA+-M2BP]PC-[WFA+-M2BP]NC), where PC = positive control and NC = negative control. Histopathological examination of liver biopsy specimen was reported according to Non-Alcoholic Steatohepatitis (NASH) Clinical Research Network Scoring System. Data for 220 cases were analyzed. The AUROC of the COI for the diagnosis of NASH was 0.65. The AUROC of the COI for the diagnosis of steatosis grade ≥2 and 3 was 0.64 and 0.53, respectively. The AUROC of the COI for the diagnosis of lobular inflammation grade ≥1, ≥2 and 3 was 0.57, 0.68 and 0.59, respectively. The AUROC of the COI for the diagnosis of hepatocyte ballooning grade ≥1 and 2 was 0.64 and 0.65, respectively. The AUROC of the COI for the diagnosis of fibrosis stage ≥1, ≥2, ≥3 and 4 was 0.61, 0.71, 0.74 and 0.84, respectively. Out of the 220 cases, 152 cases were the same 76 patients who had a repeat liver biopsy after 48 weeks of intervention. The AUROC of the change in the COI to detect improvement in steatosis, lobular inflammation, hepatocyte ballooning and fibrosis was 0.57, 0.54, 0.59 and 0.52, respectively. In conclusion, serum WFA+-M2BP was most useful for the diagnosis of significant fibrosis, advanced fibrosis and cirrhosis in NAFLD patients. However, it was less useful for differentiating NASH from non-NASH, and for diagnosis and follow-up of the individual histopathological components of NASH.