[Bone ultrasonography in kidney disease: applications and limitations].
Quantitative ultrasound (QUS) of the bone is a technique that is generating great interest among bone structure researchers because of its intrinsic features. Its safety and low cost make it an ideal technique for repeated measurements over time such as in chronic disease or when it is necessary to monitor the effects of prescribed therapies. The method was developed for the study of osteoporosis and the sites of measurement are all peripheral, including the distal diaphyses and metaphyses of the phalanges, calcaneus, radius and tibia. QUS parameters, however, cannot be used directly for the diagnosis of osteoporosis according to the WHO criteria, although many authors have shown that ultrasound parameters, particularly those of calcaneal QUS, can predict the risk of osteoporotic fractures independently of MBD. Very promising results with the use of QUS have been obtained in corticosteroid-induced osteoporosis, rheumatoid arthritis, Cushing's syndrome, cystic fibrosis, osteomalacia, thalassemia and osteopenia related to parenteral nutrition. QUS can also monitor the effectiveness of therapy in various pathological conditions. In nephrology the combined use of phalangeal QUS and biochemical markers of bone turnover allows adequate follow-up of patients on dialysis and renal transplant recipients with alterations or disorders of the bone.


Cationic/Anionic Polyelectrolyte (PLL/PGA) Coated Vesicular Phospholipid Gels (VPGs) Loaded with Cytarabine for Sustained Release and Anti-glioma Effects.
Cationic and anionic polymer-modified nanoparticles offer promising properties for the drug and gene delivery. Our study uses cationic/anionic polyelectrolyte coated vesicular phospholipid gels (VPGs) loaded with cytarabine (Ara-C) that enhance in vitro and in vivo anti-glioma effects. Sodium cholesteryl sulfate (SCS) or octadecylamine (ODA) incorporated in a phospholipids phase were used to prepare charged VPGs, and cationic ε-polylysine (PLL) coated VPGs (PLL-SCS VPGs) and anionic γ-polyglutamic acid (PGA) coated VPGs (PGA-ODA VPGs) were prepared via electrostatic interactions, respectively. The morphology, particle size, zeta potential, rheology properties, and in vitro release were then characterized. The in vitro cytotoxicity and cellular uptake were evaluated on U87-MG glioma cells. The in vivo antitumor effects were studied on BALB/c nude mice bearing a right flank U87-MG glioma model. The TEM images and physicochemical properties of cationic/anionic polyelectrolyte coated VPGs exhibited that polymers covered on the vesicular surface. The results of rheologic property analysis showed that cationic/anionic polyelectrolyte coated VPGs enhanced the viscosity of uncoated VPGs. The in vitro release experiments revealed that cationic/anionic polyelectrolyte coated VPGs kept Ara-C sustained release up to 18 days. Specially, compared with PLL-SCS VPGs, PGA-ODA VPGs demonstrated higher in vitro cytotoxicity and cellular uptake efficiency in U87-MG glioma cells, and enhanced in vivo antitumor effects when subcutaneously injected around the tumor. No severe toxicity appeared in the right flank U87-MG glioma model of BALB/c nude mice. Anionic γ-PGA coated VPGs were superior to cationic PLL coated VPGs in terms of improving the anti-glioma effect for local delivery.


Dicer-dependent endothelial microRNAs are necessary for postnatal angiogenesis.
Posttranscriptional gene regulation by microRNAs (miRNAs) is important for many aspects of development, homeostasis, and disease. Here, we show that reduction of endothelial miRNAs by cell-specific inactivation of Dicer, the terminal endonuclease responsible for the generation of miRNAs, reduces postnatal angiogenic response to a variety of stimuli, including exogenous VEGF, tumors, limb ischemia, and wound healing. Furthermore, VEGF regulated the expression of several miRNAs, including the up-regulation of components of the c-Myc oncogenic cluster miR-17-92. Transfection of endothelial cells with components of the miR-17-92 cluster, induced by VEGF treatment, rescued the induced expression of thrombospondin-1 and the defect in endothelial cell proliferation and morphogenesis initiated by the loss of Dicer. Thus, endothelial miRNAs regulate postnatal angiogenesis and VEGF induces the expression of miRNAs implicated in the regulation of an integrated angiogenic response.


Serum MicroRNA Signature Predicts Response to High-Dose Radiation Therapy in Locally Advanced Non-Small Cell Lung Cancer.
To assess the utility of circulating serum microRNAs (c-miRNAs) to predict response to high-dose radiation therapy for locally advanced non-small cell lung cancer (NSCLC). Data from 80 patients treated from 2004 to 2013 with definitive standard- or high-dose radiation therapy for stages II-III NSCLC as part of 4 prospective institutional clinical trials were evaluated. Pretreatment serum levels of 62 miRNAs were measured by quantitative reverse transcription-polymerase chain reaction array. We combined miRNA data and clinical factors to generate a dose-response score (DRS) for predicting overall survival (OS) after high-dose versus standard-dose radiation therapy. Elastic net Cox regression was used for variable selection and parameter estimation. Model assessment and tuning parameter selection were performed through full cross-validation. The DRS was also correlated with local progression, distant metastasis, and grade 3 or higher cardiac toxicity using Cox regression, and grade 2 or higher esophageal and pulmonary toxicity using logistic regression. Eleven predictive miRNAs were combined with clinical factors to generate a DRS for each patient. In patients with low DRS, high-dose radiation therapy was associated with significantly improved OS compared to treatment with standard-dose radiation therapy (hazard ratio 0.22). In these patients, high-dose radiation also conferred lower risk of distant metastasis and local progression, although the latter association was not statistically significant. Patients with high DRS exhibited similar rates of OS regardless of dose (hazard ratio 0.78). The DRS did not correlate with treatment-related toxicity. Using c-miRNA signature and clinical factors, we developed a DRS that identified a subset of patients with locally advanced NSCLC who derive an OS benefit from high-dose radiation therapy. This DRS may guide dose escalation in a patient-specific manner.


Plain packaging: a logical progression for tobacco control in one of the world's 'darkest markets'.
The Australian approach to tobacco control has been a comprehensive one, encompassing mass media campaigns, consumer information, taxation policy, access for smokers to smoking cessation advice and pharmaceutical treatments, protection from exposure to tobacco smoke and regulation of promotion. World-first legislation to standardise the packaging of tobacco was a logical next step to further reduce misleadingly reassuring promotion of a product known for the past 50 years to kill a high proportion of its long-term users. Similarly, refreshed, larger pack warnings which started appearing on packs at the end of 2012 were a logical progression of efforts to ensure that consumers are better informed about the health risks associated with smoking. Regardless of the immediate effects of legislation, further progress will continue to require a comprehensive approach to maintain momentum and ensure that government efforts on one front are not undermined by more vigorous efforts and greater investment by tobacco companies elsewhere.


Endocrine studies in a pregnancy complicated by ovarian theca lutein cysts and hyperreactio luteinalis.
Severe virilization developed in a pregnant woman in association with the occurrence of theca lutein cysts and hyperreactio luteinalis of the ovaries. At term, maternal serum levels of androstenedione (58 ng/mL) and testosterone (20 ng/mL) were elevated massively; estrone (24 ng/mL) and estradiol-17 beta (23 ng/mL) levels were increased moderately. Maternal serum levels of human chorionic gonadotropin (hCG) just before delivery, 22,276 mlU/mL, though twice the mean for normal women at term, were within the normal range. The levels of androstenedione (1.06 ng/mL) and testosterone (0.26 ng/mL) in umbilical cord serum of her normal newborn female infant were normal. The levels of androstenedione and testosterone in serum of this woman declined slowly, but progressively, during the first two months after delivery; the serum levels of androstenedione and testosterone increased substantially, however, after she was treated with hCG eight weeks postpartum. These results are suggestive that, for reasons unknown, markedly increased androgen production with theca lutein cysts and hyperreactio luteinalis in some pregnant women results ultimately because of increased ovarian sensitivity to hCG.


Inhibition of soluble epoxide hydrolase does not protect against endotoxin-mediated hepatic inflammation.
Epoxyeicosatrienoic acids (EETs) are derived from cytochrome P450-catalyzed epoxygenation of arachidonic acid and have emerged as important mediators of numerous biological effects. The major elimination pathway for EETs is through soluble epoxide hydrolase (sEH)-catalyzed metabolism to dihydroxyeicosatrienoic acids (DHETs). Based on previous studies showing that EETs have anti-inflammatory effects, we hypothesized that chronic inhibition of sEH would attenuate a lipopolysaccharide (LPS)-induced inflammatory response in vivo. Continuous dosing of the sEH inhibitors 12-(3-adamantan-1-ylureido)-dodecanoic acid (AUDA), a polyethylene glycol ester of AUDA, and 1-adamantan-1-yl-3-(5-(2-(2-ethoxyethoxy)ethoxy)-pentyl)urea resulted in robust exposure to the inhibitor and target engagement, as evidenced by significant increases in plasma EET/DHET ratios following 6 days of inhibitor treatment. However, sEH inhibitor treatment was not associated with an attenuation of LPS-induced inflammatory gene expression in the liver, and AUDA did not protect from LPS-induced neutrophil infiltration. Furthermore, Ephx2-/-mice that lack sEH expression and have significantly increased plasma EET/DHET ratios were not protected from LPS-induced inflammatory gene expression or neutrophil accumulation in the liver. LPS did have an effect on sEH expression and function, as evident from a significant down-regulation of Ephx2 mRNA and a significant shift in plasma EET/DHET ratios 4 h after LPS treatment. In conclusion, there was no evidence that increasing EET levels in vivo could modulate an LPS-induced inflammatory response in the liver. However, LPS did have significant effects on plasma eicosanoid levels and hepatic Ephx2 expression, suggesting that in vivo EET levels are modulated in response to an inflammatory signal.


[Feasibility of detecting steroid hormones in cattle feces for monitoring ovarian function].
Faecal material from four non pregnant cows was withdrawn in two-day intervalls during a four-week period. At the same time bloodsamples were taken to monitor the ovarian function. Clinical and ultrasonographical examinations were recorded in parallel. To look for the stability of the faecal steroid analogues for P4 (Progesterone) and E2 (Estradiol-17 beta) faecal material was stored at 37 degrees C and samples from that analyzed in two-day intervals. The P4 (Progesterone)-analogues values in the faeces showed good correlation to cycling ovarian functions as monitored by bloodlevels from P4, clinical examination and ultrasonography. In contrast the E2-analogues values showed no cycling pattern. One cow however showed ovarian cysts which was monitored in the P4 values for blood and faeces. Concerning the stability of the steroid analogues it could be demonstrated that the P4-values show a steady decrease with minimum values of 20 ng/g dry material. During the first four days after sampling reliable values for clinical interpretations can be found. The concentrations for E2 are very inconstant and so far not of interest. The investigation shows that the faecal steroid P4 can be used for monitoring ovarian function. It has to be analyzed within four days after sampling when stored at room temperature.


The impact of object animacy on the appraisal of causality.
The goal of this study was to examine whether animacy of objects affected the appraisal of a causal relationship when one object was observed to propel the other either immediately, after a delay, or at a distance. Participants rated the degree of causal interaction either before or after an extended experience with observing the interactions. We expected that spatial distance would have little effect when objects were seen as animate (because social interactions often span spatial gaps) and would have a degrading effect when objects were appraised as inanimate. Motion animacy appeared to attenuate the impact of a gap and to decrease initial causal judgments for direct collisions. Explicitly informing participants about the nature of the objects had a strong impact. Experience with the causal task affected ratings to a greater extent when the objects were explicitly described as nonliving rather than living.


Knockout mice: simple solutions to the problems of genetic background and flanking genes.
Inducing null mutations by means of homologous recombination provides a powerful technique to investigate gene function and has found wide application in many different fields. However, it was realized some time ago that the specific way in which such knockout mutants are generated can be confounding, making it impossible to separate the effects of the induced null mutation from those of alleles originating from the embryonic stem cell donor. In addition, effects from null mutations can be altered on different genetic backgrounds. Here we present some simple breeding strategies to test for flanking gene effects that are compatible with the recommendations of the Banbury Conference on Genetic Background in Mice and with common practices of creating and maintaining mouse knockout lines.


Condensed Tannins in the Gastrointestinal Tract of Cattle after Sainfoin (Onobrychis viciifolia) Intake and Their Possible Relationship with Anthelmintic Effects.
Condensed tannins' (CTs) fate along the digestive tract of ruminants may account for the variable efficacy of CTs against gastrointestinal nematodes. We analyzed CTs in the digesta of cattle fed sainfoin. With the acetone-butanol-HCl assay, the total CTs concentrations in the digesta were close to those in the diets (6.3 and 1.5% of DM in experiments 1 and 2, respectively); thus, CTs remained potentially largely undegraded/unabsorbed. With the thiolysis assay, CTs concentration was much higher in the abomasum (2.3% of DM; expt 1) compared with the rumen and intestines, along with higher mean size and prodelphinidins percentage, corroborating CTs efficacy reported only against Ostertagia ostertagi in the abomasum. In expt 2, the dietary levels of CTs were probably too low to demonstrate anthelmintic effects in the rumen. Overall, the level of CTs accessible to thiolysis is favored under the acidic conditions of the abomasum, which seems critical for anthelmintic activity.


Cortical symptoms described in emergency calls for patients with suspected large vessel occlusion: a descriptive analysis of 157 emergency calls.
Emergency medical dispatchers typically use the dispatch code for suspected stroke when the caller brings up one or more symptoms from the face-arm-speech triad. Paramedics and emergency department physicians are trained to suspect large vessel occlusion stroke when the stroke patient presents with hemiparesis and cortical symptoms: neglect, aphasia, and conjugate eye deviation (CED). We hypothesized that these symptoms could be evident in the emergency call. In this study, we aimed to describe common symptoms mentioned in the emergency calls for paramedic-suspected thrombectomy candidates. Secondly, we wanted to explore how the question about CED arises in the Finnish suspected stroke dispatch protocol. Our third aim was to find out if the symptoms brought up in suspected stroke and non-stroke dispatches differed from each other. This was a retrospective study with a descriptive analysis of emergency calls for patients with paramedic-suspected large vessel occlusion stroke. We listened to the emergency calls for 157 patients transported to Tampere University Hospital, a Finnish comprehensive stroke centre. Two researchers listened for symptoms brought up in these calls and filled out a pre-planned case report form. Speech disturbance was the most common symptom brought up in 125 (80%) calls. This was typically described as an inability to speak any words (n = 65, 52% of calls with speech disturbance). Other common symptoms were falling down (n = 63, 40%) and facial asymmetry (n = 41, 26%). Suspicion of stroke was mentioned by 44 (28%) callers. When the caller mentioned unconsciousness the emergency dispatcher tended to use a non-stroke dispatch code. The dispatchers adhered poorly to the protocol and asked about CED in only 57% of suspected stroke dispatches. We found CED in 12 emergency calls and ten of these patients were diagnosed with large vessel occlusion. In cases where paramedics suspected large vessel occlusion stroke, typical stroke symptoms were described during the emergency call. Speech disturbance was typically described as inability to say anything. It is possible to further develop suspected stroke dispatch protocols to recognize thrombectomy candidates from ischemic cortical signs such as global aphasia and CED.


[Histoplasmosis: not only a tropical disease].
Histoplasmosis is a fungal infection resulting from inhalation of spores from the fungus Histoplasma capsulatum; it is known to be endemic in various parts of the world, especially in North and Latin America, and can produce a spectrum of illness, from subclinical infection to progressive disseminated disease. The majority of infected persons have an asymptomatic, self-limiting illness. Clinical pneumonia occurs in those with exposure to a large number of infecting spores. Disseminated histoplasmosis usually occurs in immunosuppressed patients or in patients with chronic illness. Diagnosis is best made by visualization of yeast in tissue or by culture. In most cases, amphotericin B is the initial drug of choice, followed by one of the azoles for lifelong maintenance therapy. Itraconazole is the drug of choice for treatment of disseminated histoplasmosis in less severe cases, while fluconazole therapy for histoplasmosis is only moderately effective.


Acute pressure increase and intrarenal hemodynamics in conscious WKY and SHR rats.
Spontaneously hypertensive rats (SHR) develop proteinuria and glomerular lesions during the first year of their life. To determine whether an intrarenal hemodynamic abnormality might participate in the pathogenesis of these lesions, normotensive Wistar-Kyoto (WKY) and SHR rats were subjected to acute pressure increase with phenylephrine infusion (2.5 and 5 micrograms X kg-1 X min-1), and renal blood flow (RBF), afferent and efferent arteriolar resistances, glomerular filtration rate (GFR), and glomerular capillary hydrostatic pressure were measured or calculated. The results indicate that the two strains responded differently to the pressure rise. Thus, although the increment in renal perfusion pressure and afferent arteriolar resistance increased similarly in the two strains, efferent resistance increased only in the SHR (7.2 +/- 1.6 vs. 9.0 +/- 2.1 units, P less than 0.02) but decreased in the WKY (5.3 +/- 0.8 vs. 4.0 +/- 0.5 units, P less than 0.05). This was associated with a decreased RBF of SHR (8.31 +/- 0.71 vs. 7.22 +/- 0.57 ml X min-1 X g kidney-1, P less than 0.05) but stable RBF in WKY (7.36 +/- 0.55 vs. 7.79 +/- 0.51 ml X min-1 X g kidney-1); GFR remained unchanged in both strains. Calculated glomerular hydrostatic pressure, however, increased in the SHR (43.6 +/- 3.0 vs. 48.5 +/- 2.2 mmHg, P less than 0.05) but decreased in the WKY (33.2 +/- 2.5 vs. 28.8 +/- 1.7 mmHg, P less than 0.01). The observed higher base-line glomerular pressure in the SHR and a greater rise with phenylephrine infusion may participate in the pathogenesis of the SHR glomerular lesions and proteinuria.(ABSTRACT TRUNCATED AT 250 WORDS)


Evaluation of quantitative (90)Y SPECT based on experimental phantom studies.
In SPECT imaging of pure beta emitters, such as (90)Y, the acquired spectrum is very complex, which increases the demands on the imaging protocol and the reconstruction. In this work, we have evaluated the quantitative accuracy of bremsstrahlung SPECT with focus on the reconstruction algorithm including model-based attenuation, scatter and collimator-detector response (CDR) compensations. The scatter and CDR compensation methods require pre-calculated point-spread functions, which were generated with the SIMIND MC program. The SIMIND program is dedicated for simulation of scintillation camera imaging and only handles photons. The aim of this work was therefore twofold. The first aim was to implement simulation of bremsstrahlung imaging into the SIMIND code and to validate simulations against experimental measurements. The second was to investigate the quality of bremsstrahlung SPECT imaging and to evaluate the possibility of quantifying the activity in differently shaped sources. In addition, a feasibility test was performed on a patient that underwent treatment with (90)Y-Ibritumomab tiuxetan (Zevalin). The MCNPX MC program was used to generate bremsstrahlung photon spectra which were used as source input in the SIMIND program. The obtained bremsstrahlung spectra were separately validated by experimental measurement using a HPGe detector. Validation of the SIMIND generated images was done by a comparison to gamma camera measurements of a syringe containing (90)Y. Results showed a slight deviation between simulations and measurements in image regions outside the source, but the agreement was sufficient for the purpose of generating scatter and CDR kernels. For the bremsstrahlung SPECT experiment, the RSD torso phantom with (90)Y in the liver insert was measured with and without background activities. Projection data were obtained using a GE VH/Hawkeye system. Image reconstruction was performed by using the OSEM algorithm with and without different combinations of model-based attenuation, scatter and CDR compensations. The reconstructed images were then evaluated in terms of the accuracy of the total activity estimate in the liver insert. It was found that the activity in a large source such as the liver was estimated with a bias of around -70%, when no compensations were included in the reconstruction, whereas when compensations were included the bias obtained was between -10 and 16%. It is concluded that although the (90)Y bremsstrahlung spectrum is continuous with no pronounced peak and the count rate is low, it is possible to achieve reasonably accurate activity estimates from bremsstrahlung SPECT images if proper compensations are applied in the reconstruction. This conclusion was also confirmed by the patient study.


Deoxycholic Acid (ATX-101) for Fat Reduction.
Excess, unwanted fat in submental and other body areas has been a focus of new modalities in aesthetics. Invasive and, more recently, non-invasive modalities for removal of unwanted fat have been on an increase. ATX-101 (deoxycholic acid injection) is the only injectable drug approved in the United States and Canada for reduction of moderate or severe submental fat in adults, with ongoing trials testing its efficacy in body contouring and lipomas. It has proven efficacy in submental fat reduction with a good safety profile. This article reviews the pharmacology, mechanism of action, clinical effects and adverse effects of ATX-101. It emphasizes on careful patient selection and advises on appropriate volume administration, number of treatments, and injection technique. The literature research includes peer-reviewed articles (clinical trials or scientific reviews). Studies were identified by searching electronic databases (MEDLINE and PubMed) till December 2019 and reference lists of respective articles. Only articles published in English language were included. J Drugs Dermatol. 2021;20(11):1169-1173. doi:10.36849/JDD.3936.


Photochemical inactivation of enzymes.
The mechanisms of enzyme inactivation by ultraviolet light and visible light in the presence of sensitizing dyes are reviewed. Recent flash photolysis studies on amino acids and enzymes are summarized in terms of proposed models relating the initial photochemical reactions to permanent chemical and biological damage. The generation and reactions of singlet oxygen are discussed in connection with photodynamic processes. The photochemical results are compared with ionizing radiations, particularly pulse radiolytic methods employing radical anions as selective probes. The interrelationships between the various modes of enzyme inactivation are discussed, as well as the new information to be learned about the structure and functions of the native enzymes from selective radiation-induced alterations.


Inhibition of TYK2 and JAK1 ameliorates imiquimod-induced psoriasis-like dermatitis by inhibiting IL-22 and the IL-23/IL-17 axis.
Psoriasis is a chronic autoimmune disease affecting the skin and characterized by aberrant keratinocyte proliferation and function. Immune cells infiltrate the skin and release proinflammatory cytokines that play important roles in psoriasis. The Th17 network, including IL-23 and IL-22, has recently emerged as a critical component in the pathogenesis of psoriasis. IL-22 and IL-23 signaling is dependent on the JAK family of protein tyrosine kinases, making JAK inhibition an appealing strategy for the treatment of psoriasis. In this study, we report the activity of SAR-20347, a small molecule inhibitor with specificity for JAK1 and tyrosine kinase 2 (TYK2) over other JAK family members. In cellular assays, SAR-20347 dose dependently (1 nM-10 μM) inhibited JAK1- and/or TYK2-dependent signaling from the IL-12/IL-23, IL-22, and IFN-α receptors. In vivo, TYK2 mutant mice or treatment of wild-type mice with SAR-20347 significantly reduced IL-12-induced IFN-γ production and IL-22-dependent serum amyloid A to similar extents, indicating that, in these models, SAR-20347 is probably acting through inhibition of TYK2. In an imiquimod-induced psoriasis model, the administration of SAR-20347 led to a striking decrease in disease pathology, including reduced activation of keratinocytes and proinflammatory cytokine levels compared with both TYK2 mutant mice and wild-type controls. Taken together, these data indicate that targeting both JAK1- and TYK2-mediated cytokine signaling is more effective than TYK2 inhibition alone in reducing psoriasis pathogenesis.


Increasing prevalence of fasting hyperglycemia in adolescents aged 10-18 years and its relationship with metabolic indicators: the Korea National Health and Nutrition Examination Study (KNHANES), 2007-2018.
Pediatric diabetes is a common health burden worldwide. This study aimed to investigate the prevalence of fasting hyperglycemia in Korean youth aged 10-18 years and to evaluate its association with metabolic indicators. To assess the risk of diabetes in domestic children and adolescents, the prevalence of fasting hyperglycemia was calculated, a trend was evaluated using multi-year Korea National Health and Nutrition Examination Survey (KNHANES) data, and multivariate analysis was performed to evaluate the relationships between hyperglycemia and metabolic factors. The prevalence of fasting hyperglycemia, defined as impaired fasting glucose (fasting glucose level > 100 mg/dL and < 125 mg/dL), or diabetes mellitus (fasting glucose ≥ 126 mg/dL) was estimated in Korean teenagers. The prevalence increased from the fourth (2007-2009) to the fifth (2010-2012), sixth (2013-2015), and seventh (2016-2018) KNHANES surveys, from 5.39 to 4.79, 10.03, and 11.66 per 100 persons, respectively. In multivariate analysis, systolic blood pressure and serum triglycerides were higher in the fasting hyperglycemia group; systolic blood pressures were 109.83 mmHg and 112.64 mmHg and serum triglycerides were 81.59 mg/dL and 89.60 mg/dL in the normal blood glucose and fasting hyperglycemia groups, respectively. The prevalence of fasting hyperglycemia among children and adolescents has increased over the past decade, and this increase is potentially associated with metabolic abnormalities such as hypertension and hypertriglyceridemia. Effort is urgently required to reduce this chronic medical burden in adolescence.


Thyroid hormone induces the synthesis of a putative protein in the rat granulosa cell which stimulates progesterone release.
125I-3,5,3'-tri-iodothyronine (T3) binds specifically to a pure nuclear preparation from rat granulosa cells. A Scatchard analysis of T3 binding showed a Kd of 0.65 x 10(-9) mol/l and a Bmax of 1.57 pmol/mg DNA. The biological relevance of T3 binding to granulosa cells was evaluated by adding T3 (20 ng/incubation) to granulosa cells (1 x 10(6) cells/incubation) which greatly stimulated progesterone release. T3-stimulated progesterone release was significantly inhibited by actinomycin-D (P < 0.01) and cycloheximide (P < 0.01). T3 caused about a twofold increase in granulosa cell protein synthesis as compared with the control which was inhibited by actinomycin-D and cycloheximide. The addition of T3 to granulosa cell incubations also resulted in a more than 2.5-fold increase in mRNA. The results indicated that T3 stimulation of progesterone release is mediated via T3-induced protein(s) or TIP. TIP was located in the soluble supernatant fraction (100,000 g supernatant; 100 k sup) from T3-incubated cells but could not be detected in the 100 k sup from the control cells or LH-incubated cells. TIP was purified based on its biological activity, i.e. its addition to granulosa cell incubations stimulated progesterone release into the medium. The 100 k sup from T3-incubated granulosa cells was subjected to Sephadex G-75 gel filtration, FPLC Mono Q and FPLC Superose 6 chromatography which resulted a 273-fold purification over the starting material and a clearly homogeneous protein was obtained. SDS-PAGE of purified TIP showed it to be a 53 kDa monomer protein. Experiments conducted with radiolabelled TIP suggested internalization of TIP into the granulosa cell. The results therefore showed that T3 induces the synthesis of mRNA and proteins in rat granulosa cells and that one of the proteins is TIP which, in turn, stimulates progesterone release from the cell, suggesting thereby that this putative protein is a novel mediator of T3 function in the granulosa cell.


Unraveling substantia nigra sequential gene expression in a progressive MPTP-lesioned macaque model of Parkinson's disease.
Taking advantage of a progressive nonhuman primate model mimicking Parkinson's disease (PD) evolution, we monitored transcriptional fluctuations in the substantia nigra using Affymetrix microarrays in control (normal), saline-treated (normal), 6 days-treated (asymptomatic with 20% cell loss), 12 days-treated (asymptomatic with 40% cell loss) and 25 days-treated animals (fully parkinsonian with 85% cell loss). Two statistical methods were used to ascertain the regulation and real-time quantitative PCR was used to confirm their regulation. Surprisingly, the number of deregulated transcripts is limited at all time points and five clusters exhibiting different profiles were defined using a hierarchical clustering algorithm. Such profiles are likely to represent activation/deactivation of mechanisms of different nature. We briefly speculate about (i) the existence of yet unknown compensatory mechanisms is unraveled, (ii) the putative triggering of a developmental program in the mature brain in reaction to progressing degeneration and finally, (iii) the activation of mechanisms leading eventually to death in final stage. These data should help development of new therapeutic approaches either aimed at enhancing existing compensatory mechanisms or at protecting dopamine neurons.


Effects of artificial calculosis on rat ureter motility: peripheral contribution to the pain of ureteric colic.
The contribution of changes in ureter motility produced by a stone to the pain of ureteric calculosis is unclear. In this study we measured ureter motility as changes in intraureter pressure in anesthetized rats 1, 4, and 8 d ys after implantation of an artificial calculus (n = 33) and compared it with motility in normal (n = 8) and ligated (n = 4) ureters. Partial obstruction of the ureter by the stone produced a 478% increase in the amplitude of contractions, a 70% decrease in the rate of contractions, and a 66% decrease in the baseline pressure. The pressures reached during contractions were equivalent to those evoking nociceptive reactions in animals and humans. These changes persisted in rats that had spontaneously eliminated the stone. Complete obstruction of the ureter by the stone or by ligation abolished contractions. We conclude that the increased motility caused by a stone likely contributes to the development and maintenance of visceral pain and referred hyperalgesia in ureteric colic and to the persistence of referred hyperalgesia after elimination of the stone.


Preschool Children Exhibit Evident Compensatory Role of Internal Astigmatism in Distribution of Astigmatism: The Nanjing Eye Study.
To determine the prevalence and associated risk factors for total, corneal, and residual astigmatism and to evaluate the relations between components of astigmatism in Chinese preschool children. In the population-based, cross-sectional Nanjing Eye Study, children were measured for noncycloplegic refractive error using an autorefractor and for biometric parameters using an optical low-coherent reflectometry. Data from right eyes were analyzed to calculate the prevalence of astigmatism using various cutpoints (0.5, 1.0, and 1.5 diopters [D]) and for determining risk factors using logistic regression models. Relations between astigmatism components were assessed using Spearman correlation coefficients (ρ). Of 1817 children (mean ± SD of age: 54.8 ± 3.5 months, 54.2% male), the median (1st and 3rd quartile) of total, corneal, and residual astigmatism (vectorial difference between total and corneal astigmatism) was -0.25 (-0.50, 0), -1.06 (-1.49, -0.72), and -0.92 (-1.23, -0.62) D and their prevalence rate 1.0 D or more was 14.2%, 56.1%, and 44.2%, respectively. With-the-rule was the most common type in total astigmatism (75.2%) and in corneal astigmatism (88.2%) while against-the-rule was predominant in residual astigmatism (75.6%). A negative correlation was found between corneal J0 and internal J0 (ρ = -0.74, P < 0.001) and between corneal J45 and internal J45 (ρ = -0.87, P < 0.001). Based on compensation factor (CF), defined as the minus ratio of internal astigmatism (vectorial difference between total and anterior corneal astigmatism) and anterior corneal astigmatism, internal J0 compensated for total J0 in varying degrees (CF: 0.1-2) in 91.5% cases, while that percentage for J45 component was 77.2%. In univariate logistic regression model, older age was significantly associated with total astigmatism (odds ratio [OR] = 0.96 for per-month increase, P = 0.03), and larger axial length-corneal radius ratio was significantly associated with higher risk of residual astigmatism (OR = 2.28 for per unit increase, P = 0.03). The compensatory role of internal astigmatism on reducing corneal astigmatism was prominent in preschool children. Larger axial length-corneal radius ratio was significantly associated with higher risk of residual astigmatism.


Effect of Benzalkonium Chloride Adaptation on Sensitivity to Antimicrobial Agents and Tolerance to Environmental Stresses in Listeria monocytogenes.
Listeria monocytogenes is an important food-borne pathogen that can persist in food processing environments and thus contaminate food products. Benzalkonium chloride (BC) is a common disinfectant widely used in food industry. Selective pressure associated with exposure to BC may result in adaptation to this agent in L. monocytogenes. In this study, the effect of BC adaptation on susceptibility to antimicrobial agents and tolerance to environmental stresses, as well as the role of efflux pumps in BC adaptation were investigated in Listeria monocytogenes. Exposure of L. monocytogenes to progressively increasing concentrations of BC led to adaptation not only to BC but also to several other antimicrobial agents with different modes of action, including cefotaxime, cephalothin, ciprofloxacin, and ethidium bromide (EtBr), indicating that the disinfectant BC has the ability to select for antibiotic resistance. Reserpine, an efflux pump inhibitor, reduced minimum inhibitory concentrations (MICs) of cephalosporins, ciprofloxacin, and EtBr in BC adapted strains, indicating that efflux pumps are involved in cross-adaptation to these antimicrobial agents. Our results showed that expression levels of the efflux pump MdrL in the BC adapted strains increased significantly relative to the corresponding wild-type strains (P < 0.05), with the highest increase in one BC adapted strain named HL06BCA. Moreover, the knockout mutant HL06BCAΔmdrL showed impaired growth compared to that of HL06BCA when exposed to 2 μg/ml of BC. It suggests that efflux pump MdrL is associated with BC adaptation in L. monocytogenes. However, we did not find mdrL to be associated with cross-adaptation to cephalosporins, ciprofloxacin, and EtBr in HL06BCA. Additionally, increased sensitivity to acid, alkali, osmotic, ethanol, and oxidative stresses was observed in most strains after repeated exposure to BC. These results suggest rotation of different disinfectant is helpful to maintain high effectiveness of BC toward L. monocytogenes and ethanol and hydrogen peroxide are at least the appropriate candidates.


Nurse-sensitive indicators suitable to reflect nursing care quality: a review and discussion of issues.
To review nurse-sensitive indicators that may be suitable to assess nursing care quality. Patient safety concerns, fiscal pressures and patient expectation create a demand that healthcare providers demonstrate the quality of nursing care delivered. As a result, nurse managers are increasingly encouraged to provide evidence of nursing care quality. Nurse-sensitive indicators are being proposed as a means of meeting this need. Literature review. A review of the literature was conducted using CINAHL and MEDLINE from 2002-2011. Key search terms were nurs* and sensitive indicators, outcome measures, indicators, metrics and patient outcomes. Most of the research has examined the relationship between nursing structural variables and patient outcomes in acute care settings and have explored potential indicators for specific patient groups and nursing roles. When using nurse-sensitive indicators, issues concerning the selection, reporting and sustained use are important for nurse managers to consider. Evidence for the nurse-sensitivity of some commonly used indicators is inconsistent due to the disparity in definitions used, data collection and analysis methods. Further research on the application and implementation of these indicators is required to assist nurse managers in attempting to quantify the quality of nursing care. Nurses need to continue to strive to achieve agreement on the definitions of indicators, gather strong consistent evidence of nurse-sensitivity, resolve issues of regular data collection and consider selection, reporting and sustainment when implementing nurse-sensitive indicators. Once identified, nurse-sensitive indicators can be applied for quality improvement purposes, but consensus is required to fully realise their potential. Nurse managers need to be aware of the factors that can influence the use of indicators at unit level. Strategies need to be implemented to promote these indicators becoming integrated with routine nursing care.


Transmission measurements in air using the ESTRO mini-phantom.
The aim of this work is to study the possibility of using the ESTRO mini-phantom for transmission measurements of primary kerma in water at a point free in air. We discuss in-air measurements in general, with special attention given to in-air equivalent measurements using a water equivalent mini-phantom. The study includes four different photon energies (4, 6, 10 and 18 MV), where scoring of dose and primary kerma inside a mini-phantom in narrow beam geometry is performed with the Monte Carlo code EGS4. The results reveal that relative measurements (i.e. with and without a water absorber present) at 10 cm depth in a mini-phantom do not represent the variation of primary kerma in water at a point free in air (deviations as large as 7% at 4 MV are observed). Minimum deviations are obtained at depths somewhat larger than the depth of dose maximum. The observed deviations are due to a considerable beam hardening in the water absorber, which changes the amount of attenuation and scatter inside the mini-phantom.


Lateral Penumbra Modelling Based Leaf End Shape Optimization for Multileaf Collimator in Radiotherapy.
Lateral penumbra of multileaf collimator plays an important role in radiotherapy treatment planning. Growing evidence has revealed that, for a single-focused multileaf collimator, lateral penumbra width is leaf position dependent and largely attributed to the leaf end shape. In our study, an analytical method for leaf end induced lateral penumbra modelling is formulated using Tangent Secant Theory. Compared with Monte Carlo simulation and ray tracing algorithm, our model serves well the purpose of cost-efficient penumbra evaluation. Leaf ends represented in parametric forms of circular arc, elliptical arc, Bézier curve, and B-spline are implemented. With biobjective function of penumbra mean and variance introduced, genetic algorithm is carried out for approximating the Pareto frontier. Results show that for circular arc leaf end objective function is convex and convergence to optimal solution is guaranteed using gradient based iterative method. It is found that optimal leaf end in the shape of Bézier curve achieves minimal standard deviation, while using B-spline minimum of penumbra mean is obtained. For treatment modalities in clinical application, optimized leaf ends are in close agreement with actual shapes. Taken together, the method that we propose can provide insight into leaf end shape design of multileaf collimator.


[In vitro insecticidal activity of seed neem oil on Lutzomyia longipalpis (Diptera: Psychodidae)].
Lutzomyia longipalpis is the main vector of visceral leishmaniasis in Brazil. The objective was to evaluate the effect of oil from (Azadirachta indica) neem seeds on eggs, larvae and adults of the vector. The insects were captured in the field and kept in the laboratory at +/- 27 °C and 80% relative humidity. Five treatments with different concentrations were performed using two negative controls (distilled water and Tween 80) and a positive control. The eggs were sprayed with the oil at different concentrations and the number of hatched larvae evaluated for 10 days. Mortality of larvae was observed to pupation and adult mortality was observed after 24, 48, and 72 hours. Statistical analysis was performed by Tukey test at 5% probability. The highest oil concentration of eggs obtained 65.16 +/- 3.24% efficacy for reducing egg hatching. The test with larvae showed 67.75 +/- 2.21% efficacy at a concentration of 100 mg.mL⁻¹. In adults, the efficacy of the 100 mg.mL⁻¹ concentration was 96.64 +/- 4.11% after 24 hours. The phytochemical analysis revealed the presence of triterpenes. These results demonstrate the potential use of this oil in the control of this vector.


[On the problem of so-called ulcer in the old age (author's transl)].
By our own experiences with 471 stomach resections (gastric ulcers) we have found, that the so-called special situation of ulcer in the old age--from Spang firstly defined--has to be critically discussed. There are three groups of patients: 1. Patients with the so-called old age ulcer. 2. Patients who got their ulcer in early years of life. 3. Patients who are younger than 50 years. We mean there is no existance of a special course of this disease in elderly patients. Only the rate of complications which was most frequently to be found in the first group, differs from the other patients. For the surgeons ulcer operations have the same general problems as to be found in operations due to other reasons by elderly and old patients.


Spatially Periodic Activation Patterns of Retrosplenial Cortex Encode Route Sub-spaces and Distance Traveled.
Traversal of a complicated route is often facilitated by considering it as a set of related sub-spaces. Such compartmentalization processes could occur within retrosplenial cortex, a structure whose neurons simultaneously encode position within routes and other spatial coordinate systems. Here, retrosplenial cortex neurons were recorded as rats traversed a track having recurrent structure at multiple scales. Consistent with a major role in compartmentalization of complex routes, individual retrosplenial cortex (RSC) neurons exhibited periodic activation patterns that repeated across route segments having the same shape. Concurrently, a larger population of RSC neurons exhibited single-cycle periodicity over the full route, effectively defining a framework for encoding of sub-route positions relative to the whole. The same population simultaneously provides a novel metric for distance from each route position to all others. Together, the findings implicate retrosplenial cortex in the extraction of path sub-spaces, the encoding of their spatial relationships to each other, and path integration.


Effects of dietary inulin supplementation on the composition and dynamics of cecal microbiota and growth-related parameters in broiler chickens.
Inulin, a prebiotic, is an attractive alternative to antibiotic growth promoters in chickens. Dietary supplementation with inulin can improve growth performance, carcass yield, immune system activity, and serum biochemical parameters in chickens. A few studies investigated the impact of dietary inulin supplementation on chicken intestinal microbiota. In this study, we investigated how and why dietary supplementation with 1, 2, and 4% inulin can affect body weight gain, feed intake, food conversion rate, immunological parameters, serum biochemical parameters, and composition and dynamics of the cecal microbiota of Tegel broiler chickens using quantitative fluorescence in situ hybridization (qFISH). We showed that inulin inclusion has a negative effect on growth performance parameters before day 21 and a positive effect subsequently up to day 42. Quantitative FISH data revealed an age-dependent change in the cecal microbiota in the control broilers fed no inulin. Thus, relative abundances of Firmicutes and Actinobacteria decreased from 52.8 to 48.3% of total cells and from 8.7 to 1.4% at days 7 and 42, respectively. However, relative abundances of Bacteroidetes and Proteobacteria gradually increased from 9.3 to 26.9% of the total cells and from 10.7 to 21.1%, respectively, over the same periods. Inulin inclusion appeared to lower the relative abundances of Lactobacillus johnsonii and Bifidobacterium species at an early bird age, but it subsequently significantly (P < 0.05) increased their relative abundances. Such increases positively correlated with body weight gain of the birds, determined after day 21. Thus, dietary supplementation with inulin together with the addition of L. johnsonii and Bifidobacterium (B. gallinarum and B. pullorum) cultures at an early age may help overcome its early negative influence on growth performance. We believe that these findings can improve our knowledge on how inulin can change the intestinal microbiota of broiler chickens and help in developing an inulin feeding regime to optimize its beneficial role in chicken development.


Major bile duct injuries during laparoscopic cholecystectomy. Follow-up after combined surgical and radiologic management.
The authors provide the results of follow-up evaluation after combined surgical and radiologic management of 89 patients with major bile duct injuries during laparoscopic cholecystectomy. The incidence and mechanism of injury of major bile duct injuries during laparoscopic cholecystectomy has been clearly defined. Furthermore, a number of series have described the management of these injuries by surgical, endoscopic, and radiologic techniques with excellent short-term results. Long-term follow-up data, however, are lacking in the management of these injuries. Data were collected prospectively on 89 patients treated at a single institution with major bile duct injuries after laparoscopic cholecystectomy managed between July 1, 1990, and July 1, 1996. Patients referred with injuries underwent early percutaneous transhepatic cholangiography and biliary drainage. Based on the cholangiographic appearance and clinical situation, patients were managed by either percutaneous balloon dilatation or surgical reconstruction with a Roux-en-Y hepaticojejunostomy with transanastomotic stenting. Follow-up was obtained by personal interview during October 1996. Two patients died without an attempt at definitive therapy. Both deaths were caused by sepsis and multisystem organ failure present at the time of transfer to the authors' institution. The remaining 87 patients were managed initially by either balloon dilatation (N = 28) or surgical reconstruction (N = 59). Ten patients have not completed treatment and still have biliary stents in place. Evaluation of 25 patients completing treatment after balloon dilatation (mean follow-up, 27.8 months) showed a success rate of 64%. Evaluation of 52 patients completing treatment after surgical reconstruction (mean follow-up, 33.4 months) showed a success rate of 92%. All failures were managed successfully by either surgical reconstruction or balloon dilatation. Major bile duct injuries can be managed successfully by combined surgical and radiologic techniques. This series provides, for the first time, significant follow-up on a large number of patients with overall success rates of 64% after balloon dilatation and 92% after surgical reconstruction. The combination of surgery and balloon dilatation resulted in a successful outcome in 100% of patients treated.


'I wanted to safeguard the baby': a qualitative study to understand the experiences of Option B+ for pregnant women and the potential implications for 'test-and-treat' in four sub-Saharan African settings.
To explore what influences on engagement with Option B+ in four sub-Saharan African settings. In-depth interviews were conducted in 2015, with 22 HIV-positive women who had been pregnant since Option B+ was available, and 15 healthcare workers (HCWs) involved in HIV service delivery. Participants were purposely selected from four health and demographic surveillance sites in Malawi, Tanzania and Uganda. A thematic content analysis was conducted to investigate what influenced engagement with Option B+. Feeling 'ready' was key to pregnant women accepting antiretroviral treatment (ART) on the same day as diagnosis at antenatal clinic; this was influenced by previous knowledge of HIV-positive status, interactions with HCWs and relationship with their partners. The desire to protect their unborn infant was the main issue that motivated women to initiate treatment, temporarily over-riding barriers to starting ART. Many HCWs recognised that pressurising women into starting ART may lead them to stop treatment following delivery. However, their own responsibility to protect the infant sometimes drove HCWs to use strong persuasive techniques to initiate pregnant women onto ART as early as possible, occasionally causing women to disengage. Protecting the baby superseded feelings of unpreparedness for lifelong ART and may explain poor retention observed in Option B+ programmes. Women may benefit from more time to accept their status, and counselling on the long-term value of ART beyond the pregnancy and breastfeeding period. Strategies to promote readiness for same-day initiation of lifelong treatment are urgently needed, and may provide important lessons for universal test-and-treat implementation.


The ancestral stringent response potentiator, DksA has been adapted throughout Salmonella evolution to orchestrate the expression of metabolic, motility, and virulence pathways.
DksA is a conserved RNA polymerase-binding protein known to play a key role in the stringent response of proteobacteria species, including many gastrointestinal pathogens. Here, we used RNA-sequencing of Escherichia coli, Salmonella bongori and Salmonella enterica serovar Typhimurium, together with phenotypic comparison to study changes in the DksA regulon, during Salmonella evolution. Comparative RNA-sequencing showed that under non-starved conditions, DksA controls the expression of 25%, 15%, and 20% of the E. coli, S. bongori, and S. enterica genes, respectively, indicating that DksA is a pleiotropic regulator, expanding its role beyond the canonical stringent response. We demonstrate that DksA is required for the growth of these three enteric bacteria species in minimal medium and controls the expression of the TCA cycle, glycolysis, pyrimidine biosynthesis, and quorum sensing. Interestingly, at multiple steps during Salmonella evolution, the type I fimbriae and various virulence genes encoded within SPIs 1, 2, 4, 5, and 11 have been transcriptionally integrated under the ancestral DksA regulon. Consequently, we show that DksA is necessary for host cells invasion by S. Typhimurium and S. bongori and for intracellular survival of S. Typhimurium in bone marrow-derived macrophages (BMDM). Moreover, we demonstrate regulatory inversion of the conserved motility-chemotaxis regulon by DksA, which acts as a negative regulator in E. coli, but activates this pathway in S. bongori and S. enterica. Overall, this study demonstrates the regulatory assimilation of multiple horizontally acquired virulence genes under the DksA regulon and provides new insights into the evolution of virulence genes regulation in Salmonella spp.


Optimization of genome engineering approaches with the CRISPR/Cas9 system.
Designer nucleases such as TALENS and Cas9 have opened new opportunities to scarlessly edit the mammalian genome. Here we explored several parameters that influence Cas9-mediated scarless genome editing efficiency in murine embryonic stem cells. Optimization of transfection conditions and enriching for transfected cells are critical for efficiently recovering modified clones. Paired gRNAs and wild-type Cas9 efficiently create programmed deletions, which facilitate identification of targeted clones, while paired gRNAs and the Cas9D10A nickase generated smaller targeted indels with lower chance of off-target mutagenesis. Genome editing is also useful for programmed introduction of exogenous DNA sequences at a target locus. Increasing the length of the homology arms of the homology-directed repair template strongly enhanced targeting efficiency, while increasing the length of the DNA insert reduced it. Together our data provide guidance on optimal design of scarless gene knockout, modification, or knock-in experiments using Cas9 nuclease.


Expression in Escherichia coli and purification of human thrombopoietin.
Human thrombopoietin (TPO) has been successfully overexpressed in Escherichia coli, with an expression level of about 12% of total cellular protein. The full-length TPO gene was subcloned into the prokaryotic expression vector pKK233-2 under the control of the inducible tac promoter. The recombinant protein was produced mainly in the form of inclusion body. By efficient renaturation and one-step purification, the recombinant protein was purified to homogeneity. The specific activity and yield of recombinant TPO can reach 2 x 10(4) units/mg and 2 mg/g of wet E. coli cells respectively.


How to directly measure a Kondo cloud's length.
We propose a method to directly measure, by electrical means, the Kondo screening cloud formed by an Anderson impurity coupled to semi-infinite quantum wires, on which an electrostatic gate voltage is applied at distance L from the impurity. We show that the Kondo cloud, and hence the Kondo temperature and the electron conductance through the impurity, are affected by the gate voltage, as L decreases below the Kondo cloud length. Based on this behavior, the cloud length can be experimentally identified by changing L with a keyboard type of gate voltage or tuning the coupling strength between the impurity and the wires.


Pravastatin-induced rhabdomyolysis and purpura fulminans in a patient with chronic renal failure.
Rhabdomyolysis associated with the use of pravastatin has been demonstrated to be a rare but potentially life-threatening adverse effect of statins. Here, we report a rare case of rhabdomyolysis and purpura fulminans in a patient who had used pravastatin and developed chronic renal failure (CRF) necessitating the initiation of dialysis. We present the case of an 86-year-old man with chronic kidney disease (CKD) treated with dialysis who was admitted with back pain. He was prescribed and took pravastatin for almost 3 years to treat hyperlipidemia. He received hemodialysis therapy 7 times prior to presentation. Laboratory values included a serum creatine concentration of 6.6mg/dl and a creatinine phosphokinase (CPK) concentration of 2350IU/L. An abdominal computed tomography scan showed swollen muscles with reduced muscle density and air density in the multifidus muscle. Two days after admission, he had large, tender ecchymotic lesions and purpuric progressive skin necrosis over the back, abdomen, and upper and lower extremities. The patient died 6 days after the initial admission due to disseminated intravascular coagulation (DIC). Based on these findings and the clinical history, a diagnosis of pravastatin-induced rhabdomyolysis and purpura fulminans was made. The long-term use of statin therapy and the initiation of dialysis therapy due to ESRD, followed by a rapid onset of rhabdomyolysis within 6 days, is indicative of an elevated statin concentration. We report an extremely rare case of pravastatin-induced rhabdomyolysis and purpura fulminans with DIC in a patient with CRF.


[Mutations within the interferon sensitivity determining region in Korean patients infected with hepatitis C virus genotype 1b].
The treatment response to interferon could differ with mutations in the interferon-sensitivity-determining region (ISDR) in patients infected with hepatitis C virus (HCV) genotype-1b (HCV-Ib). We examined the pattern of ISDR mutations and analyzed whether the number of amino acid substitutions influences the treatment response to peginterferon plus ribavirin in chronic hepatitis or cirrhotic patients infected with HCV-Ib. The study population comprised 52 patients who visited Seoul Asan Medical Center and Seoul National University Bundang Hospital from January 2006 to December 2008 and who received peginterferon alpha-2a (n=37) or -2b (n=15) plus ribavirin, and whose serum was stored. We analyzed the early virologic response, end-of-treatment response, and sustained virologic response (SVR), and examined the ISDR using direct sequencing. The proportions of patients with ISDR mutation types of wild (0 mutations), intermediate (1-3 mutations), and mutant (> or =4 mutations) were 50.0%, 42.3%, and 7.7%, respectively, and the corresponding SVR rates were 63%, 50%, and 67% (p>0.05). The SVR rates were 59.4% and 50.0% in patients with <2 and > or =2 mutations, respectively (p>0.05). On univariate analysis, age was the only predictive factor for SVR (p=0.016). The pretreatment HCV RNA titer tended to be lower in those with SVR, but without statistical significance (p=0.069). The frequency of ISDR mutations was low in our cohort of Korean patients infected with HCV-Ib. Therefore, ISDR mutations might not contribute to the response to treatment with peginterferon plus ribavirin.


A new automated system for urine analysis: a simple, cost-effective and reliable method for distinguishing between glomerular and nonglomerular sources of haematuria.
To determine the ability, reliability and accuracy of a new automated system of urine analysis in differentiating glomerular from nonglomerular bleeding in the initial investigation of haematuria, and compare its efficacy with conventional phase-contrast microscopy (PCM). One hundred and six urine samples from patients in whom the final diagnosis was available were analysed using electrical flow impedance to detect, enumerate and size red blood cells in a conductive fluid (the cellfacts analyser, Microbial Systems Ltd, Coventry, UK). All the samples were also tested using a dipstick method and PCM was carried out for comparison on 45 of the 106 urine specimens. The results of cellfacts analysis were correlated with the final diagnoses to assess sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of this method; the costs were also analysed. Sixty-nine urine samples tested positive for blood on dipstick urine analysis and all these were confirmed to have red blood cells on cellfacts analysis. The remaining 37 samples were negative for blood on dipstick testing and cellfacts analysis, although seven patients had been referred with previously detected microscopic haematuria, none of whom were found to have any detectable pathology in the urinary tract on clinical examination and investigations. The remaining 30 patients were diagnosed to have urological or nephrological conditions with no haematuria. In the positive group, 20 (29%) patients were from the glomerular group, with a mean (range) red blood cell size of 4.25 (4-5.1) micrometer, and 49 (71%) from the nonglomerular group, with red blood cells of 5.47 (4.67-5.70) micrometer. These ranges overlapped at 4.67-5.1 micrometer at the decision threshold of 4.75 micrometer, the distribution of dysmorphic and eumorphic red blood cells for the glomerular group was 18 (90%) and two (10%), respectively, and for the nonglomerular group was 2 (4%) and 47 (96%), respectively. The sensitivity, specificity, PPV and NPV were 90%, 96%, 90% and 96%, respectively. Consumable and labour costs were very low. Cellfacts analysis is a simple, rapid, objective and cost-effective method for differentiating glomerular from nonglomerular urinary red blood cells, especially when few such cells are present.


Inhibitory effect of papaverine on HVJ (Sendai virus) replication in rat glioma C6 cells.
The replication of HVJ (Sendai virus) in C6 rat glial cells was found to be inhibited by treatment of the cells with papaverine, an inhibitor of cAMP phosphodiesterase, but not with cAMP or dibutyryl cAMP. In addition, cyclic GMP which often manifests a reciprocal relationship to cAMP did not counteract the inhibition of HVJ yield by papaverine. Both viral genome replication and transcription were suppressed slightly by treatment of the cells with papaverine. In the cells cultured in the presence of papaverine, the synthesis of viral proteins and their phosphorylation occurred at normal rates. Membrane immunofluorescence and cell surface immunoprecipitation showed that the viral glycoproteins HN and F0 were expressed on the cell surface of the papaverine-treated cells. Moreover, all the viral structural proteins were associated with plasma membrane isolated from the treated cells. These results indicate that papaverine treatment suppresses some part of the process of virus budding at the plasma membrane.


Antibacterial photodynamic therapy with 808-nm laser and indocyanine green on abrasion wound models.
Infections with pathogens could cause serious health problems, such as septicemia and subsequent death. Some of these deaths are caused by nosocomial, chronic, or burn-related wound infections. Photodynamic therapy (PDT) can be useful for the treatment of these infections. Our aim was to investigate the antibacterial effect of indocyanine green (ICG) and 808-nm laser on a rat abrasion wound model infected with the multidrug resistant Staphylococcus aureus strain. Abrasion wounds were infected with a multidrug resistant clinical isolate of S. aureus. ICG concentrations of 500, 1000, and 2000 μg∕ml were applied with a 450 J∕cm2 energy dose. Temperature change was monitored by a thermocouple system. The remaining bacterial burden was determined by the serial dilution method after each application. Wounds were observed for 11 days posttreatment. The recovery process was assessed macroscopically. Tissue samples were also examined histologically by hematoxylin–eosin staining. Around a 90% reduction in bacterial burden was observed after applications. In positive control groups (ICG-only and laser-only groups), there was no significant reduction. The applied energy dose did not cause any thermal damage to the target tissue or host environment. Results showed that ICG together with a 808-nm laser might be a promising antibacterial method to eliminate infections in animals and accelerate the wound-healing process.


Auditory neuropathy associated with postnatally acquired cytomegalovirus infection in a very preterm infant.
Neurosensory hearing loss is a well-known complication of antenatally acquired cytomegalovirus (CMV) infection. We here report an infant who developed auditory neuropathy after a postnatally acquired CMV infection. Infection probably occurred through ingestion of infected breast milk. Following a cochlear implant, there is normal language perception and a mildly delayed language expression at age 4. We speculate that the long-term effects of perinatal CMV infections are more dependent on the postconceptional age at which infection occurs than on whether the infection occurs antenatally or postnatally. An early acquired neonatal CMV infection in very preterm infants may therefore have long-term neurological sequelae, including auditory deficits.


Skin melanoma deaths within 1 or 3 years from diagnosis in Europe.
The steep increase in incidence of cutaneous malignant melanoma in white populations mainly applies to thin lesions with good survival suggesting overdiagnosis. Based on population-based cancer registries (CRs), we have investigated changes in aggressive melanoma, selecting only cases who died within 1 or 3 years after diagnosis in 11 European countries between 1995 and 2012. Trends in fatal cases were analysed by period of diagnosis, sex, tumour thickness, histologic subtype of the lesion, tumour site and CR with a multivariate generalised linear mixed effects model, where geographical area was considered as a random effect. We collected data on 123 360 invasive melanomas, with 5133 fatal cases at 1 year (4%) and 12 330 (10%) at 3 years. The number of fatal cases showed a 16% decrease at 1 year and 8% at 3 years between the first (1995-2000) and the last (2007-2012) period. The highest proportion of fatal cases was seen for men, older age (≥65 years), thick lesions (>1 mm), nodular melanoma, melanoma on the trunk and for poorly documented cases, lacking information about thickness and histologic subtype. The mixed-effects model showed a remarkable variability among European countries. The majority of registries showed a decreasing trend in fatal cases, but a few registries showed an opposite pattern. Trends in fatal melanoma cases, highlighting real changes in risk not related to overdiagnosis, showed a decrease in most European countries, with a few exceptions. Stronger efforts for early detection could lead to a more efficient treatment of melanoma in general.


Surgical versus medical treatment with cyclooxygenase inhibitors for symptomatic patent ductus arteriosus in preterm infants.
Patent ductus arteriosus (PDA) with significant left to right shunt in preterm infants increases morbidity and mortality. Early closure of the ductus arteriosus may be achieved pharmacologically using cyclooxygenase inhibitors or by surgery. The efficacy of both treatment modalities is well established. However, the preferred initial treatment of a symptomatic PDA in a preterm infant, surgical ligation or treatment with indomethacin, has not been well established. To compare the effect of surgical ligation of PDA vs. medical treatment with cyclooxygenase inhibitors (using indomethacin, ibuprofen, or mefenamic acid), each used as the initial treatment, on neonatal mortality in preterm infants with a symptomatic PDA. The standard search strategy of the Cochrane Neonatal Review Group was used. This included search of electronic databases: Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 2, 2007), MEDLINE (1966 - July 2007), CINAHL (1982 - July 2007), EMBASE (1980 - July 2007); and hand search of abstracts of Pediatric Academic Societies annual meetings published in Pediatric Research (1990 - April 2002) or on line from May 2002 -July 2007. No language restrictions were applied. All trials 1) using randomized or quasi-randomized patient allocation, 2) in preterm infants < 37 weeks gestational age or low-birth-weight infants (< 2500 grams) with symptomatic PDA in the neonatal period (< 28 days) and 3) comparing surgical ligation with medical treatment with cyclooxygenase inhibitors, each used as the initial treatment for closure of PDA. Assessment of methodological quality and extraction of data for included trials was undertaken independently by the authors. RevMan 4.1 was used for analysis of the data. Only one study, trial B in the report of Gersony 1983, was found eligible. No additional studies were identified in the literature searches performed in July 2007. The trial compared the effect of surgical ligation of PDA vs. medical treatment with indomethacin, each used as the primary treatment. No trials comparing surgery to other cyclooxygenase inhibitors (ibuprofen, mefenamic acid) were found. Trial B of Gersony 1983 enrolled 154 infants. The study found no statistically significant difference between surgical closure and indomethacin treatment in mortality during hospital stay, chronic lung disease, other bleeding, necrotizing enterocolitis, sepsis, creatinine level, or intraventricular hemorrhage. There was a statistically significant increase in the surgical group in incidence of pneumothorax [RR 2.68 (95% CI 1.45, 4.93); RD 0.25 (95% CI 0.11, 0.38); NNH 4 (95% CI 3, 9)] and retinopathy of prematurity stage III and IV [RR 3.80 (95% CI 1.12, 12.93); RD 0.11 (95% CI 0.02, 0.20), NNH 9 (95% CI 5, 50] compared to the indomethacin group. There was as expected a statistically significant decrease in failure of ductal closure rate in the surgical group as compared to the indomethacin group: [RR 0.04 (95% CI 0.01, 0.27); RD -0.32 (95% CI -0.43, -0.21), NNT 3 (95% CI 2, 4)]. The data regarding net benefit/harm are insufficient to make a conclusion as to whether surgical ligation or medical treatment with indomethacin is preferred as initial treatment for symptomatic PDA in preterm infants. It should be noted that three recent observational studies indicated an increased risk for one or more of the following outcomes associated with PDA ligation; chronic lung disease, retinopathy of prematurity and neurosensory impairment . It is possible that the duration of the "waiting-time" and transport to another facility with surgical capacity to have the PDA ligated could adversely affect outcomes, as could the perioperative care.


A high-throughput screening platform for Polycystic Kidney Disease (PKD) drug repurposing utilizing murine and human ADPKD cells.
Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common inherited monogenic disorders, characterized by a progressive decline in kidney function due in part to the formation of fluid-filled cysts. While there is one FDA-approved therapy, it is associated with potential adverse effects, and all other clinical interventions are largely supportive. Insights into the cellular pathways underlying ADPKD have revealed striking similarities to cancer. Moreover, several drugs originally developed for cancer have shown to ameliorate cyst formation and disease progression in animal models of ADPKD. These observations prompted us to develop a high-throughput screening platform of cancer drugs in a quest to repurpose them for ADPKD. We screened ~8,000 compounds, including compounds with oncological annotations, as well as FDA-approved drugs, and identified 155 that reduced the viability of Pkd1-null mouse kidney cells with minimal effects on wild-type cells. We found that 109 of these compounds also reduced in vitro cyst growth of Pkd1-null cells cultured in a 3D matrix. Moreover, the result of the cyst assay identified therapeutically relevant compounds, including agents that interfere with tubulin dynamics and reduced cyst growth without affecting cell viability. Because it is known that several ADPKD therapies with promising outcomes in animal models failed to be translated to human disease, our platform also incorporated the evaluation of compounds in a panel of primary ADPKD and normal human kidney (NHK) epithelial cells. Although we observed differences in compound response amongst ADPKD and NHK cell preparation, we identified 18 compounds that preferentially affected the viability of most ADPKD cells with minimal effects on NHK cells. Our study identifies attractive candidates for future efficacy studies in advanced pre-clinical models of ADPKD.


Effects of arginine-glycine-aspartic acid peptide-conjugated quantum dots-induced photodynamic therapy on pancreatic carcinoma in vivo.
Quantum dots (QDs) conjugated with integrin antagonist arginine-glycine-aspartic acid (RGD) peptides (QDs-RGD) are novel nanomaterials with a unique optical property: a high molar extinction coefficient. Previously, we have shown that QDs-RGD demonstrate a photodynamic therapy (PDT) effect as new photosensitizers for the pancreatic cancer cell line SW1990 in vitro. Here, we investigate the application of QDs-RGD in mice bearing pancreatic tumors using PDT. To ensure that more photosensitizers accumulated in tumors, QDs-RGD were injected intratumorally. After selection of an adequate dosage for injection from analyses of biodistribution images captured by an IVIS system, PDT was initiated. Three groups were created according to different PDT procedures. In group 1, mice were injected with QDs-RGD intratumorally, and an optical fiber connected to a laser light was inserted directly into the tumor. Irradiation was sustained for 20 min with a laser light (630 nm) at 100 mW/cm2. In group 2, the laser optical fiber was placed around, and not inserted into, tumors. In group 3, PDT was conducted as in group 1 but without injection of QDs-RGD. After 28 days of observation, tumors on the back of mice in group 1 grew slowly (V/V0 =3.24±0.70) compared with the control groups, whose tumors grew quickly, and the mean V/V0 reached 6.08±0.50 (group 2) and 7.25±0.82 (group 3). Histology of tumor tissues showed more necrotic tissues, more inflammatory cells, and less vascular tissue in the PDT group than those in the control groups. These results suggest that QDs-RGD-mediated PDT, with illumination using an optical fiber inserted directly into the tumor, can inhibit the growth of SW1990 tumors with high efficiency in nude mice.


Effects of decomposition on gunshot wound characteristics: under cold temperatures with no insect activity.
Information on gunshot wound characteristics has been well documented; however, there is little documented information on the effects of decomposition or environmental conditions on gunshot wound characteristics. This study was conducted in order to determine if decomposition would obscure or alter the physical surface characteristics of gunshot wounds when exposed to a low temperature environment. The study was conducted from November 2005 to January 2006 in Nova Scotia, Canada in forested and exposed environments, with air temperatures between -10 degrees C and +10 degrees C. Pigs were used as human models and were shot six times each at three different ranges (contact, 2.5 cm, and 1.5 m). Gunshot wound characteristics persisted until the wounds were covered with ice and snow, after which changes were observed. The changes were recognized as being unique to the different ranges of gunshots and it was concluded that changes due to decomposition under the conditions tested would not affect the collection and interpretation of gunshot wound evidence.


The Impact of Variation in Twin Relatedness on Estimates of Heritability and Environmental Influences.
By taking advantage of the natural variation in genetic relatedness among identical (monozygotic: MZ) and fraternal (dizygotic: DZ) twins, twin studies are able to estimate genetic and environmental contributions to complex human behaviors. Recently concerns have been raised about the accuracy of twin studies in light of findings of genetic and epigenetic changes in twins. One of the concerns raised is that MZ twins are not 100% genetically and epigenetically similar because they show variations in their genomes and epigenomes leading to inaccurate estimates of heritability. This article presents findings from a simulation study that examined the degree of bias in estimates of heritability and environmentality when the genetic and epigenetic similarity of MZ twins differs from 1.00 and when the genetic and epigenetic similarity of DZ twins differs from 0.50. The findings suggest that in the standard biometric model when MZ or DZ twin similarity differs from 1.00 or 0.50, respectively, the variance that should be attributed to genetic influences is instead attributed to nonshared environmental influences, thus deflating the estimates of genetic influences and inflating the estimates of nonshared environmental influences. Although estimates of genetic and nonshared environmental influences from the standard biometric model were found to deviate from "true" values, the bias was usually smaller than 10% points indicating that the interpretations of findings from previous twin studies are mostly correct.


Quantitative expression profiling of highly degraded RNA from formalin-fixed, paraffin-embedded breast tumor biopsies by oligonucleotide microarrays.
Microarray-based gene expression profiling is well suited for parallel quantitative analysis of large numbers of RNAs, but its application to cancer biopsies, particularly formalin-fixed, paraffin-embedded (FFPE) archived tissues, is limited by the poor quality of the RNA recovered. This represents a serious drawback, as FFPE tumor tissue banks are available with clinical and prognostic annotations, which could be exploited for molecular profiling studies, provided that reliable analytical technologies are found. We applied and evaluated here a microarray-based cDNA-mediated annealing, selection, extension and ligation (DASL) assay for analysis of 502 mRNAs in highly degraded total RNA extracted from cultured cells or FFPE breast cancer (MT) biopsies. The study included quantitative and qualitative comparison of data obtained by analysis of the same RNAs with genome-wide oligonucleotide microarrays vs DASL arrays and, by DASL, before and after extensive in vitro RNA fragmentation. The DASL-based expression profiling assay applied to RNA extracted from MCF-7 cells, before or after 24 h stimulation with a mitogenic dose of 17beta-estradiol, consistently allowed to detect hormone-induced gene expression changes following extensive RNA degradation in vitro. Comparable results where obtained with tumor RNA extracted from FFPE MT biopsies (6 to 19 years old). The method proved itself sensitive, reproducible and accurate, when compared to results obtained by microarray analysis of RNA extracted from snap-frozen tissue of the same tumor.


[Changes in transcapillary fluid exchange and the distensibility of small intestinal blood vessels upon adrenergic stimulation].
In acute experiments on cats, the mean coefficient of capillary filtration (CFC) in denervated small intestine was 0.10 ml/min/mm Hg/100 g and the stretching ability of the intestinal venous vessels was 0.21 ml/mm Hg/100 g. Infusion of noradrenaline (5 micrograms/min) reduced the CFC by 33% and the stretching ability by 24%. Blockade of vascular alpha-adrenoreceptors abolished and beta-adrenoreceptors augmented the noradrenaline effect on CFC and stretching ability. The data obtained indicate presence of both alpha- and beta-adrenoreceptors in precapillary sphincters and venous vessels of small intestine. Stimulation of the alpha-adrenoreceptors entails constriction of precapillary sphincters and decreases venous stretching ability whereas stimulation of beta-adrenoreceptors leads to opposite effects. Noradrenaline exerts main effect on alpha-adrenoreceptors of precapillary sphincters and venous vessels in the denervate intestine; alpha-blockade is followed by an obvious beta-effect in the form of relaxation of precapillary sphincters and increase of the venous stretching ability.


Targeted disruption of the IA-2beta gene causes glucose intolerance and impairs insulin secretion but does not prevent the development of diabetes in NOD mice.
Insulinoma-associated protein (IA)-2beta, also known as phogrin, is an enzymatically inactive member of the transmembrane protein tyrosine phosphatase family and is located in dense-core secretory vesicles. In patients with type 1 diabetes, autoantibodies to IA-2beta appear years before the development of clinical disease. The genomic structure and function of IA-2beta, however, is not known. In the present study, we determined the genomic structure of IA-2beta and found that both human and mouse IA-2beta consist of 23 exons and span approximately 1,000 and 800 kb, respectively. With this information, we prepared a targeting construct and inactivated the mouse IA-2beta gene as demonstrated by lack of IA-2beta mRNA and protein expression. The IA-2beta(-/-) mice, in contrast to wild-type controls, showed mild glucose intolerance and impaired glucose-stimulated insulin secretion. Knockout of the IA-2beta gene in NOD mice, the most widely studied animal model for human type 1 diabetes, failed to prevent the development of cyclophosphamide-induced diabetes. We conclude that IA-2beta is involved in insulin secretion, but despite its importance as a major autoantigen in human type 1 diabetes, it is not required for the development of diabetes in NOD mice.


Reliability and Recommended Settings for Pediatric Circumpapillary Retinal Nerve Fiber Layer Imaging Using Hand-Held Optical Coherence Tomography.
To investigate feasibility and reliability of 3-dimensional full circumpapillary retinal nerve fiber layer (cpRNFL) analysis in children, with and without glaucoma, without the use of sedation and to recommend a protocol for hand-held optical coherence tomography use. A cohort of pediatric glaucoma patients and normal children were imaged with hand-held optical coherence tomography to assess the feasibility of obtaining full cpRNFL. Two consecutive scans were acquired in a smaller sample to investigate test-retest repeatability and interassessor reproducibility. The cpRNFL thickness was assessed in four quadrants, at several visual angles from the optic nerve center. Scanning was attempted in both eyes of 90 children with pediatric glaucoma and 180 controls to investigate feasibility (mean age, 6.98 ± 4.42 years). Scanning was not possible in 68 eyes of glaucoma children mainly owing to nystagmus, unclear optical media, or high refractive errors. Where three-dimensional imaging was possible, success at obtaining full cpRNFL was 67% in children with glaucoma and 89% for controls. Seventeen children with pediatric glaucoma and 34 controls contributed to reliability analysis (mean age, 6.3 ± 3.63 years). For repeatability intraclass correlation coefficients across quadrants ranged from 0.63 to 0.82 at 4° and improved to 0.88 to 0.94 at 6°. Intraclass correlation coefficients for reproducibility were also highest at 6° (>0.97 across all quadrants). We demonstrate that acquisition and measurement of cpRNFL thickness values using 3-dimensional hand-held optical coherence tomography volumes in awake children is both feasible and reliable and is optimal at 6° from optic nerve center. Our recommended protocol provides guidance on how pediatric optic nerve pathologies are managed by clinicians.


CalpB modulates border cell migration in Drosophila egg chambers.
Calpains are calcium regulated intracellular cysteine proteases implicated in a variety of physiological functions and pathological conditions. The Drosophila melanogaster genome contains only two genes, CalpA and CalpB coding for canonical, active calpain enzymes. The movement of the border cells in Drosophila egg chambers is a well characterized model of the eukaryotic cell migration. Using this genetically pliable model we can investigate the physiological role of calpains in cell motility. We demonstrate at the whole organism level that CalpB is implicated in cell migration, while the structurally related CalpA paralog can not fulfill the same function. The downregulation of the CalpB gene by mutations or RNA interference results in a delayed migration of the border cells in Drosophila egg chambers. This phenotype is significantly enhanced when the focal adhesion complex genes encoding for α-PS2 integrin ( if), β-PS integrin (mys) and talin (rhea) are silenced. The reduction of CalpB activity diminishes the release of integrins from the rear end of the border cells. The delayed migration and the reduced integrin release phenotypes can be suppressed by expressing wild-type talin-head in the border cells but not talin-head(R367A), a mutant form which is not able to bind β-PS integrin. CalpB can cleave talin in vitro, and the two proteins coimmunoprecipitate from Drosophila extracts. The physiological function of CalpB in border cell motility has been demonstrated in vivo. The genetic interaction between the CalpB and the if, mys, as well as rhea genes, the involvement of active talin head-domains in the process, and the fact that CalpB and talin interact with each other collectively suggest that the limited proteolytic cleavage of talin is one of the possible mechanisms through which CalpB regulates cell migration.


Treatment of outpatient urinary tract infections with cinoxacin.
This paper describes an open assessment of cinoxacin in the treatment of 30 outpatients with symptomatic urinary tract infections caused by Escherichia coli, Klebsiella, Proteus mirabilis, and Enterobacter. Twenty-seven patients (90 percent) had a satisfactory clinical response and in 26 patients, there was a satisfactory microbiological response with elimination of the pathogen. Mild side effects were reported by three patients, none of whom stopped therapy. It is concluded that cinoxacin will be useful in the treatment of urinary tract disease because of the high urinary antibacterial activity produced. The relatively low incidence of side effects and convenience of twice-daily dosage should encourage good compliance by patients treated outside the hospital setting.


Expression and regulation of novel human beta-defensins in gingival keratinocytes.
This study evaluated the expression and regulation of beta-defensins DEFB-104 and the recently identified DEFB-105-14 in gingival keratinocytes. Keratinocytes from healthy subjects were exposed to cytokines, Escherichia coli lipopolysaccharide or Candida species. Total RNA was extracted and defensin expression analyzed by reverse transcription-polymerase chain reaction. Three patterns of expression were seen: no expression, constitutive expression and inducible expression. Constitutive mRNA expression was evident for DEFB-104, 107, 109, 111, and 112. DEFB-108 and 114 were induced by interleukin (IL)-1beta and Candida species. For DEFB-108 expression, synergism was observed when IL-1beta was combined with tumor necrosis factor-alpha or interferon-gamma. Downregulation of DEFB-109 occurred following treatment with Candida albicans. These findings suggest a role for multiple beta-defensins in response to oral infection. Further investigation is needed to better understand their function, both in terms of antimicrobial activities and contributions to innate and acquired immunity.


Analyses of Lysin-motif Receptor-like Kinase (LysM-RLK) Gene Family in Allotetraploid Brassica napus L. and Its Progenitor Species: An In Silico Study.
The LysM receptor-like kinases (LysM-RLKs) play a crucial role in plant symbiosis and response to environmental stresses. Brassica napus, B. rapa, and B. oleracea are utilized as valuable vegetables. Different biotic and abiotic stressors affect these crops, resulting in yield losses. Therefore, genome-wide analysis of the LysM-RLK gene family was conducted. From the genome of the examined species, 33 LysM-RLK have been found. The conserved domains of Brassica LysM-RLKs were divided into three groups: LYK, LYP, and LysMn. In the BrassicaLysM-RLK gene family, only segmental duplication has occurred. The Ka/Ks ratio for the duplicated pair of genes was less than one indicating that the genes' function had not changed over time. The BrassicaLysM-RLKs contain 70 cis-elements, indicating that they are involved in stress response. 39 miRNA molecules were responsible for the post-transcriptional regulation of 12 Brassica LysM-RLKs. A total of 22 SSR loci were discovered in 16 Brassica LysM-RLKs. According to RNA-seq data, the highest expression in response to biotic stresses was related to BnLYP6. According to the docking simulations, several residues in the active sites of BnLYP6 are in direct contact with the docked chitin and could be useful in future studies to develop pathogen-resistant B. napus. This research reveals comprehensive information that could lead to the identification of potential genes for Brassica species genetic manipulation.


Efficiency of the Therascreen® RGQ PCR kit for the detection of EGFR mutations in non-small cell lung carcinomas.
Activating mutations of the EGFR gene in lung carcinoma are associated with response to tyrosine kinase inhibitors. Therefore, a rapid, sensitive assay for mutation detection using routine pathological specimens is mandatory in clinical practice. We have compared our in-house procedure to the Therascreen® EGFR RGQ PCR kit (Qiagen). This assay, based on allele-specific amplification, is approved in the United States, in Europe, Japan and China. We first selected a series of 209 that were representative of our routine practice during the last 2years. Using our assays, EGFR mutations were detected in 36 (17.4%) of these patients (18 p.L858R mutations and 18 exon 19 deletions). All these alterations were also detected using the Therascreen® kit. In addition, this kit allowed us to detect 7 additional alterations: one exon 19 alteration (c.2239_2240TT>CC, p.L747P), 3 p.G719X mutations and 3 p.S768L mutations. In the second part of our study, we selected 81 samples that were identified as deleted for exon 19 using our assay. Eighteen different deletions were described following sequencing. All these samples were tested positive with the Therascreen® kit. The Therascreen® EGFR RGQ kit was found to be very powerful (sensitivity 100%; specificity 100%) for the detection of the most frequent EGFR alterations that are clearly associated with response to tyrosine kinase inhibitors.


[Osseous metaplasia of the endometrium as a cause of infertility. Hysteroscopic approach].
The osseous metaplasia of the endometrium is very rare cause of infertility. The hysteroscopic approach is a effective method to recover the fertility in most of the cases in a short period. The case of a infertile woman who was previous studied and has indications for laparoscopic and hysteroscopic approach. The infertility workup showed a intracavitary calcification on ultrasonography. During the surgery, the diagnosis of endometriosis and osseous metaplasia of the endometrium were done and treated with histologic confirmation. The patient conceived in her second spontaneous cycle. The hysteroscopy is the first approach in the treatment and should be doing with laparoscopy for the detection of other causes of infertility.


Muscarinic receptor-mediated calcium signaling in spiral ganglion neurons of the mammalian cochlea.
Using indo-1 microspectrofluorometry, we examined the effects of cholinergic agonists on the concentration of intracellular Ca(2+) ions ([Ca(2+)](i)) in spiral ganglion neurons, isolated from rat cochleae at different stages of post-natal development (from P3 to P30). Extracellular application of acetylcholine (ACh) or carbamylcholine generated a rapid and transient increase in [Ca(2+)](i). The ACh concentration-response curve indicated an apparent dissociation constant (K(d)) of 8 microM and a Hill coefficient of 1.0. Removing extracellular free Ca(2+) did not suppress the ACh-induced Ca(2+) responses suggesting an intracellular Ca(2+)-release mechanism. When we compared the cholinergic response at different stages of postnatal development, there were no significant differences on the aspect of the Ca(2+) response and the percentage of responsive neurons, which ranged between 50 and 65% per cochlear preparation. The application of muscarine triggered reversible Ca(2+) responses similar to those observed with ACh, with an apparent K(d) of 10 microM and a Hill coefficient of 1.0. The cholinergic-induced Ca(2+)pirenzepine. Nicotine (10 to 100 microM) did not evoke Ca(2+) responses and the nicotinic antagonist curare (10 microM) did not block the ACh-evoked responses. The present study is the first direct demonstration of functional muscarinic receptors (mAChRs) in spiral ganglion neurons. These mAChRs activated by the cholinergic lateral efferent system may participate in the regulation of the electrical activity of the afferent auditory fibers contacting the inner hair cells.


Hypertrophic branchial myopathy with uniform predominance of type 1 fibres. Case report.
A 17-year-old Japanese girl presented with left masseteric hypertrophy. She had a habit of chewing only on the affected side. Histochemical study of the masseter muscle showed myopathic changes characterised by uniform type 1 fibres with gross hypertrophy, increases of internal nuclei and split fibres, and endomysial fibrous tissue. This condition could be compatible with hypertrophic branchial myopathy, because it was confined to the masseter muscle and so cannot be classified into any known type of systemic myopathy.


Bilayer Interactions among Unsaturated Phospholipids, Sterols, and Ceramide.
Using differential scanning calorimetry and lifetime analysis of trans-parinaric acid fluorescence, we have examined how cholesterol and cholesteryl phosphocholine (CholPC) affect gel-phase properties of palmitoyl ceramide (PCer) in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1,2-dioleyol-sn-glycero-3-phosphocholine (DOPC) bilayers. By 2H NMR, we also measured fluid-phase interactions among these lipids using deuterated analogs of POPC, PCer, and cholesterol. The PCer-rich gel phase in POPC bilayers (9:1 molar ratio of POPC to PCer) was partially and similarly dissolved (and thermostability decreased) by both cholesterol and CholPC (sterol was present equimolar to PCer, or in fourfold excess). In DOPC bilayers (4:1 DOPC/PCer molar ratio), CholPC was much more efficient in dissolving the PCer-rich gel phase when compared to cholesterol. This can be interpreted as indicating that PCer interaction with POPC was stronger than PCer interaction with DOPC. PCer-CholPC interactions were also more favored in DOPC bilayers compared to POPC bilayers. In the fluid POPC-rich phase, cholesterol increased the order of the acyl chain of d2-PCer much more than did CholPC. In DOPC-rich fluid bilayers, both cholesterol and CholPC increased d2-PCer acyl chain order, and the ordering induced by CholPC was more efficient in DOPC than in POPC bilayers. In fluid POPC bilayers, the ordering of 3-d1-cholesterol by PCer was weak. In summary, we found that in the gel phase, sterol effects on the PCer-rich gel phase were markedly influenced by the acyl chain composition of the fluid PC. The same was true for fluid-phase interactions involving the sterols. Our results further suggest that PCer did not display high affinity toward either of the sterols used. We conclude that the nature of unsaturated phospholipids (POPC versus DOPC) in bilayers has major effects on the properties of ceramide gel phases and on sterol-ceramide-phospholipid interactions in such complex bilayers.


Drought tolerance response of high-yielding soybean varieties to mild drought: physiological and photochemical adjustments.
Soybean is a crop of agronomic importance that requires adequate watering during its growth to achieve high production. In this study, we determined physiological, photochemical and metabolic differences in five soybean varieties selected from the parental lines of a nested association mapping population during mild drought. These varieties have been described as high yielding (NE3001, HY1; LD01-5907, HY2) or drought tolerant (PI518751; HYD1; PI398881, HYD2). Nevertheless, there has been little research on the physiological traits that sustain their high productivity under water-limited conditions. The results indicate that high-yielding varieties under drought cope with the shortage of water by enhancing their photoprotective defences and invest in growth and productivity, linked to a higher intrinsic water use efficiency. This is the case of the variety N-3001 (HY1), with a tolerance strategy involving a faster transition into the reproductive stage to avoid the drought period. The present study highlights the role of the physiological and biochemical adjustments of various soybean varieties to cope with water-limited conditions. Moreover, the obtained results underscore the fact that the high phenotypic plasticity among soybean phenotypes should be exploited to compensate for the low genetic variability of this species when selecting plant productivity in constrained environments.