[Effect of a fragment of pBR322 plasmid in the region of tetracycline resistance gene on the stability of this plasmid].
A fragment destabilizing the pBR322 plasmid has been localized in the region of pBR322 tet gene promoter. On the basis of stability analysis of pBR322 derivatives comprising the modified region of tet gene, we deduced that it is the DNA sequence localized at the beginning of tet gene in the region of HindIII splitting site that ensures the plasmid destabilization, and not tet gene product or the active transcription of this region. The destabilizing effect of this fragment of the plasmid is manifested both in cis and in trans. Possible molecular mechanisms of the phenomenon are discussed.


[Is the pneumoperitoneum minimally invasive during laparoscopic colonic surgery?].
The importance of laparoscopic colonic surgery has increased considerably in the past decade. However, a minimally invasive operation with induction of pneumoperitoneum does not imply a minimally invasive anaesthesia. The haemodynamic effects of intraperitoneal carbon dioxide insufflation depend an the extent of intraabdominal pressure elevation, severity of preexisting cardiopulmonary diseases, alterations of arterial PCO (2) and pH, volume state of the patient and co-medications. In addition, positioning of the patient for laparoscopic colonic surgery and endocrinological reactions during and after induction of pneumoperitoneum may significantly affect systemic and pulmonary haemodynamics. Intraabdominal operations may impair respiratory function independent from anaesthesia. Preoperative evaluation of the high risk patient is of utmost importance. Assessment of expiratory PCO (2), extended cardiopulmonary monitoring and maintenance of intraabdominal pressure in the range of 5 - 7 mmHg are recommended during laparoscopic colonic surgery.


TNFα-blockade stabilizes local airway hyperresponsiveness during TLR-induced exacerbations in murine model of asthma.
Viral infections are a common cause of asthma exacerbation. These maladies are sometimes complicated by bacterial infections. Toll-like receptors (TLRs) are in the forefront of our microbial defence, with TLR3 responding to viral and TLR4 to bacterial stimulation. The present study was designed to evaluate the effect of concomitant TLR3 and TLR4 stimulation in a murine model of allergic asthma.BALB/c mice were stimulated intranasally with a combination of poly(I:C) and LPS activating TLR3 and TLR4, respectively. This resulted in the development of airway hyperresponsiveness (AHR) in the proximal part of the lung, along with signs of neutrophilic inflammation. Analysis of the bronchioalveolar lavage fluid (BALF) revealed a marked increase in TNFα. In contrast, the allergic airway inflammation induced by ovalbumin administration to sensitized mice caused AHR in the whole lung along with an increase in eosinophils and lymphocytes in the BALF and lung.When poly(I:C) + LPS were given to mice with an ongoing allergic airway inflammation induced by ovalbumin, the AHR was further increased in the peripheral lung and neutrophils appeared together with eosinophils and lymphocytes in the BALF and lung. Treatment with the TNFα-blocking antibody infliximab blunted the AHR increase, without affecting the cells influx in BALF.To conclude; a combined TLR3- and TLR4-stimulation, representing a concomitant viral and bacterial infection, causes an AHR that is further exaggerated during an ongoing allergic inflammation. The airway stabilizing effect of infliximab indicates the possible future use of TNFα blockade in treatment of microbial induced exacerbations of allergic asthma.


Intraoperative electrical stimulation of the brain in patients with obsessive-compulsive neurosis.
Twenty patients with intractable obsessive-compulsive neurosis were operated under local anesthesia. Each patient had a lesion produced in 1 of the 4 brain targets: anterior internal capsule, rostral cingulum, middle cingulum, and genu of the corpus callosum. Before destructive permanent lesions were produced, the target area was stimulated electrically. Stimulation gave subjective or objective reactions in 30% of the patients: diminished anxiety, 3 patients; increased anxiety, 1 patient, and motor responses, 2 patients. None experienced an obsessive reaction to stimulation. The clinical effect of surgery was usually good.


Attachment style and subjective motivations for sex.
The relation of attachment style to subjective motivations for sex was investigated in an Internet survey of 1999 respondents. The relations of attachment anxiety and avoidance to overall sexual motivation and to the specific motives for emotional closeness, reassurance, self-esteem enhancement, stress reduction, partner manipulation, protection from partner's negative affect and behavior, power exertion, physical pleasure, nurturing one's partner, and procreation were explored. As predicted, attachment anxiety was positively related to overall sexual motivation and to all specific motives for sex, with the exception of physical pleasure. Avoidance was negatively related to emotional closeness and reassurance as goals of sex and positively related to manipulative use of sex but minimally related to most other motives. Sexual passion was positively related to attachment anxiety and negatively related to avoidance, and anxiety was related to the maintenance of passion over time, whereas avoidance was related to loss of passion over time.


Photophysical properties of a 1,2,3,4,5,6-hexasubstituted fullerene derivative.
The photophysical properties of a novel 1,2,3,4,5,6-hexasubstituted fullerene derivative (1) are examined in this study. In addition to the ground state absorption spectrum of 1, we report its triplet-triplet absorption spectrum and molar extinction coefficient (Deltae(T-T)), as well as the triplet quantum yield (PhiT), lifetime (tauT), and energy (ET). The saturation of a single six-member ring on the fullerene cage results in significant changes in the triplet state properties as compared to that of pristine C60. The triplet-triplet absorption spectrum shows a hypsochromic shift in long wavelength absorption, and both the triplet state lifetime and the triplet quantum yield are decreased. The triplet energy was found to be similar to that of C60. In addition, the quantum yield (PHI(delta)) of singlet oxygen generated by 1 was calculated and is found to be significantly less than in the case of C60.


Ruthenium terpyridine complexes containing a pyrrole-tagged 2,2'-dipyridylamine ligand-synthesis, crystal structure, and electrochemistry.
Ruthenium(II) terpyridine complexes containing the pyrrole-tagged 2,2'-dipyridylamine ligand PPP (where PPP stands for N-(3-bis(2-pyridyl)aminopropyl)pyrrole with the general formula [Ru(tpy)(PPP)X](n+) (1, X = Cl(-); 2, X = H(2)O; 3, X = CH(3)CN; tpy = 2,2':6',2"-terpyridine) have been synthesized and characterized by (1)H NMR, IR, UV-vis, mass spectrometry, and elemental analysis. 1 and 2 have been structurally characterized by X-ray crystallography. Both 1 and 2 were successfully immobilized onto glassy carbon electrode via anodic oxidation of the pyrrole moiety on the PPP ligand to give stable and highly electroactive polymer films. Cyclic voltammetric studies of 1 in acetonitrile revealed a Ru(III)/Ru(II) couple at 0.4 V vs Cp(2)Fe(+/0) initially, but another redox couple resulting from chloride substitution by acetonitrile developed at E(1/2) = 0.82 V upon repetitive potential scan. This ligand substitution was induced by the acidic local environment caused by the release of protons during pyrrole polymerization. The electropolymerization of 2 in aqueous medium allowed the observation of the formation of Ru(IV)═O species in polypyrrole film. As the film grew thicker, the size of the Ru(III)/(/)Ru(II) couple (E(1/2) = 0.8 V vs SCE at pH 1) of poly[Ru(tpy)(PPP)(OH(2))](n+) increased accordingly, whereas the growth of the Ru(IV)/Ru(III) couple (E(1/2) = 0.89 V vs SCE at pH 1) leveled off after the film had reached a certain thickness. The Pourbaix diagram of the E(1/2) of the Ru(III) /Ru(II) and Ru(IV)/Ru(III) couples vs pH of the electrolyte medium has been obtained. The resulting poly[Ru(tpy)(PPP)(OH(2))](n+) film is electrocatalytically active toward the oxidation of benzyl alcohol.


Evaluation of Ginger (Zingiber officinale Roscoe) Bioactive Compounds in Increasing the Ratio of T-cell Surface Molecules of CD3+CD4+:CD3+CD8+ In-Vitro.
The potential ability of ginger bioactive compounds in increasing the ratio of T-cell surface molecules of CD3+CD4+:CD3+CD8+ was investigated using dual tagging FITC and PE of monoclonal antibody anti-human with its fluorescence measured by flow cytometer. Oleoresin was extracted using sinkhole distillation technique. Its components namely, gingerol in fraction-1, shogaol in fraction 2 and zingeron in fraction-3 were separated by column vacuum chromatography method. The doses of oleoresin, gingerol, shogaol, and zingeron tested were 50, 100,150, 200, and 250 μg/ml. Lymphocytes (2x106 cell/ml) from human peripheral blood were isolated using ficoll density gradient technique, and cultured in the presence of the compounds in RPMI-1640 medium and phytohemaglutinin (PHA) mitogen for 96 h under normal conditions. Percentages of T-cell surface molecules (CD4+ and CD8+) were determined using dual-tagging FITC and PE fluorescents labeled on monoclonal antibody anti human. The fluorescence-labeled bands on the T-cell surface molecules were counted using flow cytometer. The experiment revealed that oleoresin and its three fractions increased the percentage of CD3+CD4+. The compound in fraction 3 of oleoresin at 200 μg/ml increased by the highest percentage of CD3+CD4+ of 9%, but slightly decreased the percentage of CD3+CD8+. These ginger bioactive compounds increased the ratio of CD3+CD4:CD3+CD8+ T-cells with the highest increment of 30% from effects of 200 μg/ml fraction 3 of oleoresin. This in vitro finding revealed that ginger bioactive compounds potentially increased cellular and humoral immune response. Further clinical studies are needed to confirm the benefits of these ginger bioactive compounds as a potential functional food for testing on HIV infected patients.


Assessment of the usefulness of skin phototype and skin color as the parameter of cutaneous narrow band UVB sensitivity in psoriasis patients.
Many reports have been released to assess skin types, skin colors and cutaneous sensitivity to broad band UVB or UVA. This study was performed to investigate the usefulness of skin type and skin color as the parameter of narrow band UVB (NBUVB) sensitivity. The minimal erythema dose (MED) of 40 psoriasis patients was investigated by irradiating several doses ranging from 200 to 1500 mJ/cm2. Before phototesting, the skin color of buttock was measured with a tristimulus colorimeter. The median and mode value of MED of NBUVB was 950 mJ/cm2. Skin type was well correlated with the MED and there was a significant relationship between the L* value and MED, but not for the a* and b* values. The MED value of NBUVB in our study is a basic data to set the phototherapy protocol. Our result showed that skin type and L* value might be useful for predicting the sensitivity to NBUVB irradiation.


Build-up mechanisms determining the topology of mutualistic networks.
The frequency distribution of the number of interactions per species (i.e., degree distribution) within plant-animal mutualistic assemblages often decays as a power-law with an exponential truncation. Such a truncation suggests that there are ecological factors limiting the frequency of supergeneralist species. However, it is not clear whether these patterns can emerge from intrinsic features of the interacting assemblages, such as differences between plant and animal species richness (richness ratio). Here, we show that high richness ratios often characterize plant-animal mutualisms. Then, we demonstrate that exponential truncations are expected in bipartite networks generated by a simple model that incorporates build-up mechanisms that lead to a high richness ratio. Our results provide a simple interpretation for the truncations commonly observed in the degree distributions of mutualistic networks that complements previous ones based on biological effects.


Inhibition by erythroid differentiation factor (activin A) of P-glycoprotein expression in multidrug-resistant human K562 erythroleukemia cells.
The K562/VCR cell line, exhibiting acquired multidrug resistance (MDR) with increased expression of a cell surface glycoprotein (P-glycoprotein), was isolated from human erythroleukemia K562 cells. Various compounds that induced erythroid differentiation of K562 cells were tested for their effects on growth and differentiation of these K562/VCR cells. Sodium butyrate, hemin, 1-beta-D-arabinofuranosylcytosine, and erythroid differentiation factor (EDF) induced erythroid differentiation of K562/VCR cells as well as K562 cells. The MDR of K562/VCR cells was partly overcome by treatment with EDF but not with the other inducers. Expression of P-glycoprotein by K562/VCR cells was measured by radioimmunoassay using MRK16 monoclonal antibody. Results showed that EDF caused down-regulation of P-glycoprotein in K562/VCR cells, whereas the other inducers did not cause its down-regulation. Thus, in addition to inducing erythroid differentiation, EDF enhanced the sensitivity of K562/VCR cells to multidrugs and suppressed expression of P-glycoprotein. These results suggest that differentiation inducers may be useful in chemotherapy of leukemic MDR cells.


Isoprenoid biosynthesis in dandelion latex is enhanced by the overexpression of three key enzymes involved in the mevalonate pathway.
Latex from the dandelion species Taraxacum brevicorniculatum contains many high-value isoprenoid end products, e.g. triterpenes and polyisoprenes such as natural rubber. The isopentenyl pyrophosphate units required as precursors for these isoprenoids are provided by the mevalonate (MVA) pathway. The key enzyme in this pathway is 3-hydroxy-methyl-glutaryl-CoA reductase (HMGR) and its activity has been thoroughly characterized in many plant species including dandelion. However, two enzymes acting upstream of HMGR have not been characterized in dandelion latex: ATP citrate lyase (ACL), which provides the acetyl-CoA utilized in the MVA pathway, and acetoacetyl-CoA thiolase (AACT), which catalyzes the first step in the pathway to produce acetoacetyl-CoA. Here we isolated ACL and AACT genes from T. brevicorniculatum latex and characterized their expression profiles. We also overexpressed the well-characterized HMGR, ACL and AACT genes from Arabidopsis thaliana in T. brevicorniculatum to determine their impact on isoprenoid end products in the latex. The spatial and temporal expression profiles of T. brevicorniculatum ACL and AACT revealed their pivotal role in the synthesis of precursors necessary for isoprenoid biosynthesis in latex. The overexpression of A. thaliana ACL and AACT and HMGR in T. brevicorniculatum latex resulted in the accumulation of all three enzymes, increased the corresponding enzymatic activities and ultimately increased sterol levels by ~5-fold and pentacyclic triterpene and cis-1,4-isoprene levels by ~2-fold. Remarkably high levels of the triterpene precursor squalene were also detected in the triple-transgenic lines (up to 32 mg/g root dry weight) leading to the formation of numerous lipid droplets which were observed in root cross-sections. We could show the effective expression of up to three transgenes in T. brevicorniculatum latex which led to increased enzymatic activity and resulted in high level squalene accumulation in the dandelion roots up to an industrially relevant amount. Our data provide insight into the regulation of the MVA pathway in dandelion latex and can be used as a basis for metabolic engineering to enhance the production of isoprenoid end products in this specialized tissue.


Scabies presenting as cutaneous nodules or malar erythema: reports of patients with scabies surrepticius masquerading as prurigo nodularis or systemic lupus erythematosus.
Scabies surrepticius is a unifying term that represents non-classical presentations of scabies mite infestation. A patient with scabies surrepticius is described: a man with scabies masquerading as prurigo nodularis. The 91-year-old man had metastatic prostate cancer and presented with diffuse pruritic nodules. Prurigo nodularis was suspected; however, the biopsy revealed scabies mites in the stratum corneum. He was successfully treated with topical permethrin 5% cream and oral ivermectin. In addition, the features of a woman with scabies mimicking systemic lupus erythematosus are summarized. The 47-year-old woman had idiopathic thrombocytopenic purpura and presented with malar erythema and a positive antinuclear antibody (titer 1:320). A diagnosis of systemic lupus erythematous was entertained until skin scraping and mineral oil preparation revealed scabies mites; she was successfully treated with oral ivermectin. In conclusion, Sarcoptes scabiei infestation can present with atypical clinical morphology and an absence of classical lesions such as burrows conventionally distributed in the interdigital web spaces, volar wrists, periumbilical area, or genitalia. Scabies surrepticius is a term that has been designated to describe these unusual presentations. Prurigo nodularis and systemic lupus erythematosus can be added to the litany of conditions masquerading as scabies and are included amongst the guises of scabies surrepticius.


Health-related quality of life after urinary diversion: continent diversion with the Kock pouch compared with ileal conduit. A questionnaire study.
A questionnaire study was carried out to compare the health-related quality of life after continent urinary diversion using a Kock pouch with that following ileal conduit diversion. 76 patients completed the questionnaire: 26 patients had a continent and 50 an ileal conduit diversion, median follow-up was 0.9 year (range 0.5-4.4) and 4.9 years (range 2.4-14.7), respectively. The type of operation carried significant relation to the presence of leakage distress associated with urine leakage; 78% among conduit patients versus 50% among continent diversion patients experienced distress. Physical contact between patient and partner was reduced significantly less following continent diversion: 16% against 43% among patients with conduit diversion. Global satisfaction with life was high and similar in the two groups. Continent urinary diversion appears to be superior to conventional ileal conduit diversion.


DOI and alpha-methylserotonin: comparative vascular and nonvascular smooth muscle effects and central 5-hydroxytryptamine2 receptor affinities.
Both alpha-methylserotonin and 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) are agonists at 5-hydroxytryptamine2 (5-HT2) receptors. The present study compared these agonists for their binding affinities at the high- and low-affinity states of the 5-HT2 receptor and for their contractile activities in certain smooth muscle preparations. Both agonists contracted the rat aorta and rat jugular vein, tissues possessing 5-HT2 receptors, and contraction was blocked by ketanserin. However, alpha-methylserotonin produced greater maximal response (80-90% maximum response to serotonin) than DOI. In the rat jugular vein, the calculated dissociation constant of DOI (-log Kb = 7.7) corresponded well with its affinity for [3H]ketanserin- (pKi = 7.5) but not [125I]DOI- (pKi = 8.6) radiolabeled sites. This might suggest that binding to the agonist low-affinity state of the 5-HT2 receptor is more relevant to vascular agonist activity. alpha-Methylserotonin was slightly more potent than serotonin in contracting the rat aorta but not the jugular vein, whereas DOI was more potent in the jugular vein than in the aorta. In the jugular vein but not the aorta, the relative potency of these agents corresponded well with their relative affinities for either the [3H]ketanserin- or the [125I]DOI-labeled 5-HT2 receptor.(ABSTRACT TRUNCATED AT 250 WORDS)


The occurrence of l-lactate dehydrogenase in the inner mitochondrial compartment of pig liver.
Although pig represents a model species in biomedical research including studies dealing with liver patho-physiology, some aspects of liver metabolism need to be addressed. In particular, whether and how pig mitochondria can metabolize l-lactate remains to be established. We show here that pig liver mitochondria (PLM) possess their own l-lactate dehydrogenase (mL-LDH). This was shown both via immunological analysis and by assaying photometrically the L-LDH reaction in solubilised PLM. The mL-LDH reaction shows hyperbolic dependence on the substrate concentration, it is inhibited by oxamate and proves to differ from the cytosolic activity (cL-LDH), as revealed by the difference found in both pH profiles and temperature dependence of m- and cL-LDH. Titration experiments with digitonin show that mL-LDH is restricted in mitochondrial inner compartment. In agreement with the above findings, three genes in Sus scrofa genome encoded for L-LDH subunits which are predicted to have mitochondrial localization, as investigated by Target P 1.1 and PredSL analysis.


Study of the influence of auxiliary substances on the partition coefficients of some substituted phenylcarbamic acid derivatives.
In the course of the drug form development of newly synthetized potential drugs the influence of the auxiliary substances, glycerol (GL), propylene glycol (PG), and sorbitol (SO) in concentration of 5, 10, 15 and 20%, on the partition coefficient (P') of some phenylcarbamic acid derivatives was studied. Because of the great variations of this influence, attempts were made to find optimal additive for the further processing. The obtained values of P' show that an optimal additive for the drug H+B seems to be SO while in the case of the substances XIX M and XIX the optimal additives are PG or GL and SO, respectively.


Endothelial cell interactions and integrins.
Adhesion molecules on the endothelium and leukocytes are involved in mediating cell-cell adhesion, which is an initial step in the leukocyte migration response. These adhesion molecules, some of which are present constitutively and others that can be up-regulated in response to chemotactic and proinflammatory stimuli, regulate the trafficking of leukocytes across the vascular endothelial barrier. A basic mechanism of interaction between the endothelium and leukocytes involves CD11/CD18 integrins, which bind to members of the immunoglobulin super gene family-intercellular adhesion molecule (ICAM)-1 and ICAM-2. The interaction between CD11/CD18 integrins and ICAM-1 is required for leukocyte migration. The other important adhesive interaction between leukocytes and the endothelium involves binding of selectins to their endothelium carbohydrate counterreceptors on leukocytes. The selectin-mediated binding is involved in leukocyte "rolling," whereas CD11/CD18 integrins are responsible for strengthening and stabilizing the leukocyte adhesion. This review summarizes the current literature on leukocyte adhesion molecules, in particular, the endothelial cell interactions mediated by the CD11/CD18 integrins.


Resolving the so-called "probabilistic paradoxes in legal reasoning" with Bayesian networks.
Examples of reasoning problems such as the twins problem and poison paradox have been proposed by legal scholars to demonstrate the limitations of probability theory in legal reasoning. Specifically, such problems are intended to show that use of probability theory results in legal paradoxes. As such, these problems have been a powerful detriment to the use of probability theory - and particularly Bayes theorem - in the law. However, the examples only lead to 'paradoxes' under an artificially constrained view of probability theory and the use of the so-called likelihood ratio, in which multiple related hypotheses and pieces of evidence are squeezed into a single hypothesis variable and a single evidence variable. When the distinct relevant hypotheses and evidence are described properly in a causal model (a Bayesian network), the paradoxes vanish. In addition to the twins problem and poison paradox, we demonstrate this for the food tray example, the abuse paradox and the small town murder problem. Moreover, the resulting Bayesian networks provide a powerful framework for legal reasoning.


Coomassie brilliant blue R-250 as a highly sensitive pre-stain for immediate visualization of human serum proteins on polyacrylamide gel disc electrophoresis and raising monospecific antibodies.
We hereby describe electrophoretic analysis of normal human serum pre-stained with Coomassie Brilliant Blue R-250 (CBB). The proposed method was optimized by studying as many as 450 disc electrophoretic separations and 30 variables. The method when compared with the post-electrophoretic staining by Amido Black (AB) revealed that the pre-stained discs were intensely well defined and resolved within 2 hours with a transparent gel. Gels stained with AB, despite a prolonged destaining, showed a residual dye retention making identification of the faint components difficult. Protein eluted from the CBB pre-stained gels retained its purity and immunoreactivity and conjugates of the two prototype proteins namely the albumin and the transferrin produced high-titre monospecific antisera in immunized rabbits.


Informed consent in pediatric research.
Pediatric drug research is gradually becoming more and more accepted as the norm for assessing whether a drug is safe and efficacious for infants and children. The process of informed consent and assent for these trials presents a major challenge. The aim of this review is to map historical, ethical and legal aspects relevant to the challenges of informed consent in the setting of pediatric drug research. The impact of age, level of maturity and life circumstances on the process of obtaining informed consent as well as the relations between consent and assent are discussed. There appears to be a lack of regulatory clarity in the area of pediatric clinical trials; while numerous statements have been made regarding children's rights to autonomy and their ability to care for themselves and for younger ones, the ever changing status of adolescence is still difficult to translate to informed consent. This may delay scientific and clinical advancement for children who are at the very junction of being independent and not needing parental permission. Obtaining consent and assent for pediatric clinical trials is a delicate matter, as both parent and child need to agree to participate. The appropriate transfer of information to guardians and the children, especially concerning potential risks and benefits, is at the heart of informed consent, as it serves to protect both patient and physician. As many adults lack health literacy, one must ensure that guardians receive relevant information at a level and in forms they can understand regarding the trials their children are asked to participate in.


Micronuclei in genotoxicity assessment: from genetics to epigenetics and beyond.
Micronuclei (MN) are extra-nuclear bodies that contain damaged chromosome fragments and/or whole chromosomes that were not incorporated into the nucleus after cell division. MN can be induced by defects in the cell repair machinery and accumulation of DNA damages and chromosomal aberrations. A variety of genotoxic agents may induce MN formation leading to cell death, genomic instability, or cancer development. In this review, the genetic and epigenetic mechanisms of MN formation after various clastogenic and aneugenic effects on cell division and cell cycle are described. The knowledge accumulated in literature on cytotoxicity of various genotoxins is precisely reflected and individual sensitivity to MN formation due to single gene polymorphisms is discussed. The importance of rapid MN scoring with respect to the cytokinesis-block micronucleus assay is also evaluated.


FDG PET/CT and mediastinal nodal metastasis detection in stage T1 non-small cell lung cancer: prognostic implications.
We aimed to compare the prognoses of patients with pathologically true negative (P-TN) N2 and PET/CT false negative (FN) results in stage T1 non-small cell lung cancer (NSCLC). Our institutional review board approved this retrospective study with a waiver of informed consent. The study included 184 patients (124 men and 60 women; mean age, 59 years) with stage T1 NSCLC who underwent an integrated PET/CT and surgery. After estimating the efficacy of PET/CT for detecting N2 disease, we determined and compared disease-free survival (DFS) rates in three groups (P-TN [n = 161], PET/CT FN [n = 12], and PET/CT true positive [TP, n = 11]) using the Kaplan-Meier analysis and log-rank test. Pathologic N2 disease was observed in 23 (12%) patients. PET/CT had an N2 disease detection sensitivity of 48% (11 of 23 patients), a specificity of 95% (153 of 161), and an accuracy of 89% (164 of 184). The 3-year DFS rate in the PET/CT FN group (31%, 95% confidence interval [CI]; 13.6-48.0%) was similar to that of the TP group (16%, 95% CI; 1.7-29.5%) (p = 0.649), but both groups had significantly shorter DFS rates than the P-TN group (77%, 95% CI; 72.0-81.2%) (p < 0.001). The PET/CT shows a high specificity, but low sensitivity for detecting N2 disease in stage T1 NSCLC. Patients with PET/CT FN N2 disease have survival rates similar to PET/CT TP N2 disease patients, which are both substantially shorter than the survival rate of P-TN patients.


Comparison of Plasma and Intrapulmonary Concentrations of Nafithromycin (WCK 4873) in Healthy Adult Subjects.
The nafithromycin concentrations in the plasma, epithelial lining fluid (ELF), and alveolar macrophages (AM) of 37 healthy adult subjects were measured following repeated dosing of oral nafithromycin at 800 mg once daily for 3 days. The values of noncompartmental pharmacokinetic (PK) parameters were determined from serial plasma samples collected over a 24-h interval following the first and third oral doses. Each subject underwent one standardized bronchoscopy with bronchoalveolar lavage (BAL) at 3, 6, 9, 12, 24, or 48 h after the third dose of nafithromycin. The mean ± standard deviation values of the plasma PK parameters after the first and third doses included maximum plasma concentrations (Cmax) of 1.02 ± 0.31 μg/ml and 1.39 ± 0.36 μg/ml, respectively; times to Cmax of 3.97 ± 1.30 h and 3.69 ± 1.28 h, respectively; clearances of 67.3 ± 21.3 liters/h and 52.4 ± 18.5 liters/h, respectively, and elimination half-lives of 7.7 ± 1.1 h and 9.1 ± 1.7 h, respectively. The values of the area under the plasma concentration-time curve (AUC) from time zero to 24 h postdosing (AUC0-24) for nafithromycin based on the mean or median total plasma concentrations at BAL fluid sampling times were 16.2 μg · h/ml. For ELF, the respective AUC0-24 values based on the mean and median concentrations were 224.1 and 176.3 μg · h/ml, whereas for AM, the respective AUC0-24 values were 8,538 and 5,894 μg · h/ml. Penetration ratios based on ELF and total plasma AUC0-24 values based on the mean and median concentrations were 13.8 and 10.9, respectively, whereas the ratios of the AM to total plasma concentrations based on the mean and median concentrations were 527 and 364, respectively. The sustained ELF and AM concentrations for 48 h after the third dose suggest that nafithromycin has the potential to be a useful agent for the treatment of lower respiratory tract infections. (This study has been registered at ClinicalTrials.gov under registration no. NCT02453529.).


Co-existence of severe coarctation of the aorta and aortic valve stenosis in adulthood: a case report.
Aortic coarctation is a congenital heart disease that usually presents and is treated in the childhood. The aortic coarctation is often associated with concomitant cardiac pathologies, such as aortic stenosis and bicuspid aortic valve. We report the case of a 56-year-old man, admitted in our cardiologic unit, for sudden onset of chest pain, dyspnea and syncope. Aortic coarctation with aortic valve stenosis was diagnosed. The aortic valve was successfully replaced in the first stage; the coarctation had to be treated in a second time.


Trends in fatal occupational injuries in Latino/a workers relative to other groups, North Carolina 2000-2017.
Latino/a workers may experience higher fatal occupational injury rates than non-Latino/a workers. In North Carolina, the Latino/a population more than doubled between 2000 and 2017. We examined fatal occupational injuries among Latino/a and non-Latino/a workers in North Carolina over this period. Information on fatal occupational injuries was abstracted from records of the North Carolina Office of the Chief Medical Examiner and the death certificate records held by the North Carolina Office of Vital Records. Estimates of the working population were derived from the decennial census and American Community Survey. Estimates of annual rates of fatal occupational injury for the period January 1, 2000 to December 31, 2017 were derived for Latino/a workers and compared to Black and White workers not identified as Latino/a. Over the study period, 1,783 fatal occupational injuries were identified among non-Latino/a workers and 259 fatal occupational injuries among Latino/a workers in North Carolina. The majority of fatal occupational injuries among Latino/a workers occurred among males employed in construction and agriculture. While the fatal occupational injury rate among Latino/a workers declined over the study period, the rate among Latino/a workers was higher than among non-Latino/a White and Black workers; moreover, fatal occupational injury rates for Latino/a workers trended upwards during the most recent years of the study period. Latino/a workers in North Carolina have the highest fatal occupational injury rate of any race/ethnicity group.


[Diagnostic value of pan-cavography in obstructed central venous return of intrinsic causes].
The authors, after having analyzed the clinical patterns and the causes producing cava syndrome, propose a sequential diagnostic approach to it, whose course makes use of radiological procedures more or less invasive, and of endoscopic and histobioptic techniques. Among radiological investigations pan-cavography has allowed the authors to formulate the diagnosis of limited venous return because of intrinsic causes, conferring so an aetiological connotation and a physiopathological correlation indispensable for a correct and rapid therapy of the uncommon "intrinsic cava syndrome" of two patients, suffering respectively from "idiopathic thrombosis of superior vena cava, concerning also the beginning of inferior vena cava" and from "Fiedler cardiomyopathy".


Effects of Tamsulosin and Tolterodine on double J stent-related symptoms: A double-blind, randomized, placebo-controlled trial.
Ureteral double J stent are routinely applied for urologic patients although stent-related symptoms are common. Several attempts have been reported to minimize these symptoms. To compare Tolterodine, Tamsulosin, and placebo effects on double J stent-related symptoms. In all, 125 patients (82 males and 43 females) with double J stent were randomly divided into three groups (group 1, n: 42, group2, n: 40 and group 3, n: 43). Each patient randomly received one pack of drug in different colors by a nurse unaware of the content to take Tamsulosin 0.4 mg before sleep (MODALUSINE), Tolterodine 2 mg twice a day or placebo once daily (capsules filled with starch): group 1 received placebo, group 2 Tamsulosin and group 3 Tolterodine for 1 month in a double-blind manner. Ureteral stent-related morbidity indices which analyzed include urinary symptom, pain, general health, quality of work and sex scores. All of indices measured by Ureteral Symptom Score Questionnaire for first and fourth weeks after drug consumption and the first week after double J stent removal (labeled as w1, w4, and w5, respectively). The mean age was 44.8 years (range: 15-83 years). There was no statistically significant difference in background characteristics between groups (p value > 0.05). The most important and statistically significant results were Tolterodine-reduced urinary symptom score (p value = 0.001) and improved general health score (p value = 0.007) of the fourth week. The pain score in groups of Tamsulosin and Tolterodine significantly reduced between weeks 4 and 1 and 5 and 1 (both with the p value < 0.05), but in other indices, there was no significant difference between them. According to our results, we suggest Tolterodine to minimize stent-related urinary symptom and improve general health in patients with double J stent.


Eucheuma cottonii Sulfated Oligosaccharides Decrease Food Allergic Responses in Animal Models by Up-regulating Regulatory T (Treg) Cells.
In the present study, the anti-food allergy activity of Eucheuma cottonii sulfated oligosaccharide (ESO) was investigated. ESO was obtained by enzymatic degradation and purified by column chromatography. RBL-2H3 cells and BALB/c mouse model were used to test the anti-food allergy activity of ESO. The effects of ESO on the regulatory T (Treg) cells and bone marrow-derived mast cells (BMMCs) were investigated by flow cytometry. The results of in vivo assay showed that ESO decreased the levels of mast cell protease-1 and histamine and inhibited the levels of specific IgE by 77.7%. In addition, the production of interleukin (IL)-4 and IL-13 was diminished in the ESO groups compared to the non-ESO-treated group. Furthermore, ESO could up-regulate Treg cells by 22.2-97.1%. In conclusion, ESO decreased the allergy response in mice by reducing basophil degranulation, up-regulating Treg cells via Forkhead box protein 3 (Foxp3), and releasing IL-10. ESO may have preventive and therapeutic potential in allergic disease.


A simple isothermal DNA amplification method to screen black flies for Onchocerca volvulus infection.
Onchocerciasis is a debilitating neglected tropical disease caused by infection with the filarial parasite Onchocerca volvulus. Adult worms live in subcutaneous tissues and produce large numbers of microfilariae that migrate to the skin and eyes. The disease is spread by black flies of the genus Simulium following ingestion of microfilariae that develop into infective stage larvae in the insect. Currently, transmission is monitored by capture and dissection of black flies and microscopic examination of parasites, or using the polymerase chain reaction to determine the presence of parasite DNA in pools of black flies. In this study we identified a new DNA biomarker, encoding O. volvulus glutathione S-transferase 1a (OvGST1a), to detect O. volvulus infection in vector black flies. We developed an OvGST1a-based loop-mediated isothermal amplification (LAMP) assay where amplification of specific target DNA is detectable using turbidity or by a hydroxy naphthol blue color change. The results indicated that the assay is sensitive and rapid, capable of detecting DNA equivalent to less than one microfilaria within 60 minutes. The test is highly specific for the human parasite, as no cross-reaction was detected using DNA from the closely related and sympatric cattle parasite Onchocerca ochengi. The test has the potential to be developed further as a field tool for use in the surveillance of transmission before and after implementation of mass drug administration programs for onchocerciasis.


Influenza A H1N1 pdm09-associated myocarditis during zanamivir therapy.
A 9-year-old girl developed influenza A H1N1 pdm09-associated myocarditis and pericarditis 2 days after starting zanamivir therapy. The virus was detected in the respiratory tract but not in the serum or pericardial effusion. The virus sampled from the respiratory tract had normal susceptibility to neuraminidase inhibitors. Although no differences in interferon-γ, interleukin (IL)-1β, and tumor necrosis factor-α were observed between the plasma and pericardial effusion, some inflammatory cytokines or chemokines (IL-6 and IL-8) and vascular endothelial growth factor were remarkably elevated in the pericardial effusion compared with the plasma. This suggested that the influenza virus, after infecting the respiratory tract, affected the myocardium, causing myocarditis to gradually develop, which might have been followed by an autoreactive pericarditis causing increased pericardial effusion. Therefore, influenza-associated myocarditis should be considered when influenza patients have respiratory and cardiac involvement, even during treatment with a neuraminidase inhibitor.


Accuracy diagrams: a novel way to illustrate uncertainty of estimated GFR.
Most studies that validate GFR equations present accuracy results stratified by measured GFR (mGFR; diagnostic correctness) or by estimated GFR (eGFR; diagnostic predictiveness) only, without a clear distinction in interpretation. The accuracy of a GFR equation is normally reported in percent (e.g. P30), but is often misinterpreted when stratified by eGFR. The aim of the study was to develop new accuracy measures and diagrams that allow straightforward interpretations and illustrations of the uncertainty in eGFR in clinical practice. We applied quantile regression to the distribution of estimation errors for two creatinine-based GFR equations, LM-REV and CKD-EPI, in a clinical cohort (n = 3495) referred for GFR measurement (plasma clearance of iohexol). Measures of bias and precision and accuracy intervals (AIs) were expressed in mL/min/1.73 m2. Diagrams with AIs were chosen as a novel way to present the error margin in eGFR at a pre-specified certainty level. It was shown that creatinine-based equations are still quite inaccurate in that large estimation errors could not be ruled out with satisfactory certainty. As an example, the 75% AI for the most accurate equation, LM-REV, was approximately ±10 mL/min/1.73 m2 at eGFR = 45 mL/min/1.73 m2, whereas it ranged between -13 and +20 mL/min/1.73 m2 at eGFR = 90 mL/min/1.73 m2. Accuracy intervals presented in diagrams can be used to illustrate the uncertainty of eGFR. Future validation studies should assess the variability in the predictiveness of eGFR across populations and clinical settings using tools and performance measures that are easy to interpret.


Role of α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor regulation in stress-induced pain chronification.
Persistent postsurgical pain is a serious issue in public health, which has received increased interest in recent years. Previous studies have reported that psychological factors promote the development of chronic postsurgical pain. However, it is unclear how chronification of postsurgical pain occurs. The α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor (AMPA) phosphorylation in the central nervous system plays a critical role in synaptic plasticity and contributes to central sensitization and chronic pain development. Here, we discuss the role of AMPA receptor regulation in stress-induced pain chronification after surgery.


COVID-19 has no impact on disease activity, progression and cognitive performance in people with multiple sclerosis: a 2-year study.
Sequelae of COVID-19 in people with multiple sclerosis (PwMS) have not been characterised. We explored whether COVID-19 is associated with an increased risk of disease activity, disability worsening, neuropsychological distress and cognitive dysfunction during the 18-24 months following SARS-COV-2 infection. We enrolled 174 PwMS with history of COVID-19 (MS-COVID) between March 2020 and March 2021 and compared them to an age, sex, disease duration, Expanded Disability Status Scale (EDSS), and a line of treatment-matched group of 348 PwMS with no history of COVID-19 in the same period (MS-NCOVID). We collected clinical, MRI data and SARS-CoV2 immune response in the 18-24 months following COVID-19 or baseline evaluation. At follow-up, PwMS also underwent a complete neuropsychological assessment with brief repeatable battery of neuropsychological tests and optimised scales for fatigue, anxiety, depression and post-traumatic stress symptoms. 136 MS-COVID and 186 MS-NCOVID accepted the complete longitudinal evaluation. The two groups had similar rate of EDSS worsening (15% vs 11%, p=1.00), number of relapses (6% vs 5%, p=1.00), disease-modifying therapy change (7% vs 4%, p=0.81), patients with new T2-lesions (9% vs 11%, p=1.00) and gadolinium-enhancing lesions (7% vs 4%, p=1.00) on brain MRI. 22% of MS-COVID and 23% MS-NCOVID were cognitively impaired at 18-24 months evaluation, with similar prevalence of cognitive impairment (p=1.00). The z-scores of global and domain-specific cognitive functions and the prevalence of neuropsychiatric manifestations were also similar. No difference was detected in terms of SARS-CoV2 cellular immune response. In PwMS, COVID-19 has no impact on disease activity, course and cognitive performance 18-24 months after infection.


The diagnosis of internal mammary node metastases of breast cancer.
It is important to know the status of internal mammary lymph node (IMN) in estimating the prognosis in patients with breast cancer and/or planning the treatment. In the first series of this study, the diagnostic value of internal mammary lymphoscintigraphy (IMLS) and that of parasternal sonography (PS) were evaluated in 57 patients. The overall diagnostic accuracy of IMLS was 72% and that of PS, 84%, compared with IMN metastases confirmed on a biopsy. In the second series, 23 patients who underwent extended radical mastectomy and were found to have IMN metastases, were examined for the sizes and locations of metastatic IMN. Minute metastatic lymph nodes not detectable by IMLS or PS were found in 9 (39%) of the patients. However, the incidence of metastases to the first and/or second intercostal spaces was 97% for the patients. We concluded that a biopsy of the first and second intercostal spaces is useful in providing the information of IMN status.


Analysis of variability in length of sleep state bouts reveals memory-free sleep subcomponents consistent among primary insomnia patients.
The statistical distributions of bout lengths for the different (macro) sleep states (wake, N1, N2, N3, and REM sleep) are essential to understanding whether any memory-free subcomponent ("micro state") is involved in the organization of sleep. Micro state detection can be prevented by the fusion of data including various sources of variability, in particular by the differences in sleep architecture between individuals, along sleep time (or nighttime), or between different nights. In this analysis, a mathematical model of sleep was adopted to disentangle these features and advance the understanding of the dynamics and mechanisms of sleep and its states. The analysis involved 116 primary insomnia patients taking placebo before going to bed and undergoing polysomnography for one night. The individual sequences of macro sleep states had been previously modeled with a mixed-effect nonhomogeneous modified Markov chain model, from which individual conditional probability distributions for the bout durations were derived in this analysis as functions of sleep time. The probability distributions, affected by neither subject, night-time, nor multiple-night pooling, substantially changed at ¼ and ¾ sleep time, had modified exponential shape, and were best described as the sum of one to four exponentials, depending on the sleep state. The time constants and proportions of bouts contributing to each exponential were similar in the different subjects, changing over sleep time. Variability in bout durations thus indicated the presence of multiple memory-free sleep subcomponents whose mean residence times and access probabilities could be identified and shown to be consistent among the studied subjects. NEW & NOTEWORTHY We present a new methodology for deriving, from polysomnography data, the individual conditional probability for the duration of the bouts of wake, N1, N2, N3, and REM sleep. We evaluated the variability of this probability within and between primary insomnia patients and along sleep time. The multiexponential shapes of the probability distributions within the individuals revealed memory-free mechanisms and sleep subcomponents with consistent features in the studied population.


Use of primary prevention implantable cardioverter-defibrillators in a population-based cohort is associated with a significant survival benefit.
Underuse of implantable defibrillators has been previously noted in patients at risk for sudden cardiac death, as well as for survivors of sudden cardiac death. We sought to determine the utilization rates in a primary prevention implantable cardioverter-defibrillator (ICD)-eligible population and mortality in this group compared with a group that had undergone implantation of this therapy. A retrospective cohort of patients from April 1, 2006, to December 31, 2009, was used to define a primary prevention ICD-eligible population. Two groups were compared on the basis of ICD implantation (no-ICD versus ICD). The primary outcome measure was mortality. Of the 717 patients found to be potentially eligible for a primary prevention ICD, 116 (16%) were referred. The remaining cohort of 601 patients were compared with an existing cohort of primary prevention ICD patients (n=290). A significant survival benefit was associated with primary prevention ICD implantation (hazard ratio, 0.46; 95% CI [0.33-0.64]; P<0.0001). When adjusted for prespecified variables known to be associated with overall mortality and propensity score, a similar survival benefit was seen (hazard ratio, 0.59; 95% CI [0.40-0.87]; P=0.01). Appropriate ICD therapy occurred in 26% of those in the ICD group, during a mean follow-up of 2.7 years. A significant mortality benefit was observed for patients who underwent primary prevention ICD implantation compared with those who did not. Vigilance is required to ensure that patients eligible for primary prevention ICDs are appropriately referred and assessed to allow such patients to benefit from this life-saving therapy.


H7N9 and H5N1 avian influenza suitability models for China: accounting for new poultry and live-poultry markets distribution data.
In the last two decades, two important avian influenza viruses infecting humans emerged in China, the highly pathogenic avian influenza (HPAI) H5N1 virus in the late nineties, and the low pathogenic avian influenza (LPAI) H7N9 virus in 2013. China is home to the largest population of chickens (4.83 billion) and ducks (0.694 billion), representing, respectively 23.1 and 58.6% of the 2013 world stock, with a significant part of poultry sold through live-poultry markets potentially contributing to the spread of avian influenza viruses. Previous models have looked at factors associated with HPAI H5N1 in poultry and LPAI H7N9 in markets. However, these have not been studied and compared with a consistent set of predictor variables. Significant progress was recently made in the collection of poultry census and live-poultry market data, which are key potential factors in the distribution of both diseases. Here we compiled and reprocessed a new set of poultry census data and used these to analyse HPAI H5N1 and LPAI H7N9 distributions with boosted regression trees models. We found a limited impact of the improved poultry layers compared to models based on previous poultry census data, and a positive and previously unreported association between HPAI H5N1 outbreaks and the density of live-poultry markets. In addition, the models fitted for the HPAI H5N1 and LPAI H7N9 viruses predict a high risk of disease presence for the area around Shanghai and Hong Kong. The main difference in prediction between the two viruses concerned the suitability of HPAI H5N1 in north-China around the Yellow sea (outlined with Tianjin, Beijing, and Shenyang city) where LPAI H7N9 has not spread intensely.


Combined rapid-steam hydrolysis and organosolv pretreatment of mixed southern hardwoods.
Various types of pretreatments are used for biomass conversion of woods. The major objective of most pre treatments is to increase the susceptibility of cellulose and lignocellulose material to acid and enzymatic hydrolysis. In this study, southern mixed hardwoods were pretreated by combined rapid steam hydrolysis (RASH) and organosolv methods. It was found that the major factor in the pretreatment was the RASH temperatures. The organosolv temperature had only a minor effect on the reactivity of the final product. The enzymatic rate studies indicated that the RASH process helps in increasing the accessibility of cellulose to enzymatic hydrolysis and increased the amount of soluble lignin While the organosolv process only removed solubilized lignin. Another effect of the combined treatment was the decreasing of the enzymatic rate relative to a single RASH pretreatment. All hemicellulose is lost during these pretreatments. Three alcohols (methanol, ethanol, and butanol) were studied using a combined RASH organosolv process. At lower temperatures there were small differences between the alcohols; however, at higher temperatures all alcohols were equally effective. At longer RASH times, the percentage of glucose in the final product, as well as the amount of solubilized lignin, increased. However, the longer RASH times led to a decrease in enzymatic rates, Organosolv residence time studies of 15, 30, and 45 minutes displayed little effect on the product. Various wood-to-solvent ratios and water-to-alcohol ratios had very little effect on the yield of products. The stability of RASH treated material be fore organosolv process was studied under various storage conditions. The storage conditions had no apparent effect on the product.


Medical packaging: towards global harmony.
The goal of one uniform set of rules that can be interpreted and applied to all medical device packages, wherever they are produced or utilised, is closer to becoming reality. Just a few more operational steps are required for completion. The shape of the future as well as current and future test methods are outlined.


Clinical Evaluation of Short (6 mm) and Longer Implants Placed Side by Side in Posterior Partially Edentulous Area: A 3-Year Observational Study.
Short implants have been proposed as an alternative solution for the rehabilitation of atrophic posterior region. To compare the clinical outcomes between 6 mm short implants and conventional implants placed under similar conditions of bone quality and occlusal loading. Nine patients received atone 6 mm implant and one standard-length (8 mm length or longer) implants in a total of 10 partially edentulous areas. Implants were left submerged for 3-6 months healing period and the screw-retained splinted prostheses were delivered. When the provisional or final restoration was placed, and at each year after loading, standardized intraoral radiograph was taken for themarginal bone level (MBL) changes around the implants. Subsequently, the patients were recalled for the clinical examination evaluating the implant survival, sulcus bleeding index, suppuration, and the incidence of prosthetic complications at every 6 months after the definitive crown delivery. The observation period was continued to 3 years (mean follow-up was 3.4 ± 0.3 years) after functional loading. Nine patients (10 short implants and 10 standard length implants) were selected in this study. Cumulative survival rates of the short implants and standard-length implants were 100% in both groups, and no biological and prosthetic complication were found in 3 years observation period. Cortical bone thickness of implant insertion sites was 1.39 ± 0.45 mm at short implants and 1.38 ± 0.69 mm at standard-length implants, and trabecular bone computed tomography values of implant insertion sites was 424.1 ± 290.1 at short implants and 410.9 ± 267.9 at standard-length implants. The MBL changes were -0.30 ± 0.71 mm at short implants and -0.19 ± 0.78 mm at standard-length implants at 3 years follow-up visit. No significant difference was found in the average of MBL changes between implant length. Within the limits of this study, it can be concluded that 6 mm short implants in a posterior edentulous region showed excellent results compared with conventional implants.


Gonadal steroid regulation of substance P (SP) and SP-encoding messenger ribonucleic acids in the rat anterior pituitary and hypothalamus.
Substance P-like immunoreactivity (SP-LI) is present in the rat anterior pituitary (AP) and in hypothalamic neurons that may be involved in the control of AP secretion and/or reproductive function. The presence of multiple SP-encoding mRNAs and tachykinin peptides and their regulation by steroid hormones were examined in APs and hypothalami from normal, gonadectomized, and steroid-treated male and female rats. SP-encoding mRNAs were identified by nuclease protection assays of RNA, and tachykinin peptides were identified by combined HPLC-RIA of tissue extracts, beta- and gamma-preprotachykinin (PPT) mRNAs and SP, neurokinin A, and neuropeptide gamma peptides were identified in the AP. The alpha-, beta-, and gamma-PPT mRNAs and SP, neurokinin A, neuropeptide gamma, neuropeptide K, and neurokinin B peptides were present in hypothalamic tissue. Previous studies have established that in the AP, SP is differentially regulated by gonadal steroids; estrogen decreases and androgen increases AP SP. Steroid effects were further analyzed in experiments using RIAs to measure SP levels in the AP and median eminence (ME) of steroid- and oil-treated gonadectomized rats. To assess whether steroids alter steady state PPT mRNA levels and presumably SP synthesis in these tissues, potential effects on AP and hypothalamic SP-encoding mRNAs were determined. Ovariectomized rats treated for 10 days with estradiol benzoate showed a 50% decrease in AP SP and a 90% decrease in AP beta- and gamma-PPT mRNAs compared to ovariectomized oil-treated controls. Estradiol benzoate replacement had no effect on SP levels in the isolated ME, but did cause a 50% increase in alpha-, beta-, and gamma PPT mRNAs in the hypothalamus. Although there was no significant effect of testosterone propionate on AP SP levels in castrated males, 10 days of testosterone propionate replacement did cause a significant increase in beta- and gamma PPT mRNAs in the AP. No androgen effects were seen on either ME SP or hypothalamic SP-encoding mRNAs. These data demonstrate that estrogen up-regulates SP-encoding mRNAs in the hypothalamus, whereas it down-regulates SP-encoding mRNAs in the pituitary. These results implicate SP and other tachykinins derived from the SP gene as steroid-regulated modulators of AP secretion and possibly reproductive function.


Nocturnal asthma: role of snoring and obstructive sleep apnea.
In this study, we documented the clinical features of patients who had obstructive sleep apnea (OSA) and coexisting asthma and assessed the safety of nocturnal nasal continuous positive airway pressure (nCPAP) therapy on the stability of asthma. Nine patients (8 men and 1 woman) with asthma and OSA confirmed on all-night sleep study were studied. All patients suffered from frequent nocturnal asthma attacks, resulting in hospitalizations and respiratory arrests in 3. All patients had symptoms of heavy snoring, nocturnal choking, frequent sleep arousals, and excessive daytime sleepiness. They recorded their daily peak expiratory flow rates (PEFR) in the mornings and evenings, before and after bronchodilator in three 2-wk periods consisting of control, nCPAP, and control. During the period of nCPAP therapy, all patients recorded improvement in their PEFR. The mean prebronchodilator and postbronchodilator PEFR for the 9 patients were significantly higher during nCPAP therapy than during both control periods. This study confirms that nCPAP therapy can be used safely in treating patients with OSA and coexisting asthma. Furthermore, nCPAP treatment improves the asthma control and, in particular, the nocturnal attacks in this group of patients. These results also suggest that recurrent upper airway obstruction and snoring may be important triggering mechanisms of nocturnal asthma attacks.


Regulation of protein activities by phosphoinositide phosphates.
Phosphoinositide phosphates (PIPs) correspond to phosphorylated derivatives of phosphatidylinositol (PI). Despite their relatively low abundance in the plasma membrane, PIPs play a crucial role as precursors of second messengers and are themselves important signaling and targeting molecules. Indeed, modulation of levels of PIPs affects, for example, cortical actin organization, membrane dynamics, and cell migration. The focus of this review is on selected interesting targets of PIPs. Those proteins that bind PIPs and are involved in regulation of actin assembly, actin membrane linkage, and actin contractility are discussed, as well as those that are involved in signaling, such as small GTPases, protein kinases, and phosphatases, or in regulation of membrane dynamics.


Bradycardia, Renal Failure, Atrioventricular Nodal Blockade, Shock and Hyperkalaemia Syndrome Involving Digoxin Toxicity: A Case Report.
Bradycardia, renal failure, atrioventricular nodal blockade, shock and hyperkalemia (BRASH) syndrome is named after the pentad of symptoms experienced by patients with this clinical entity, and is propagated via a synergistic mechanism. Herein, we describe a case of an 81-year-old male who presented with bradycardia, dyspnoea on exertion, and confusion. He was also initially found to be in cardiogenic shock. In a setting of elevated digoxin levels, acute renal failure and hyperkalemia, he was diagnosed with BRASH syndrome. Prompt interventions of continuous renal replacement therapy and digoxin antibody administration were performed to treat this patient. His renal function improved and his hyperkalemia and bradycardia resolved over the course of 4 days, and the patient was discharged to a subacute rehabilitation facility after stabilization. BRASH syndrome is a clinical entity requiring prompt diagnosis for life-saving treatment, including renal replacement therapy, vasoactive medications, transvenous pacing, and reversing agents, when appropriate.


Effects of whey protein isolate on body composition, lipids, insulin and glucose in overweight and obese individuals.
The health benefits currently associated with increased dairy intake may be attributable to the whey component of dairy proteins. The present study evaluated the effects of whey protein supplementation on body composition, lipids, insulin and glucose in comparison to casein and glucose (control) supplementation in overweight/obese individuals for 12 weeks. The subjects were randomised to whey protein, casein or glucose supplementation for 12 weeks according to a parallel design. Fasting blood samples and dual-energy X-ray absorptiometry measurements were taken. Seventy men and women with a mean age of 48.4 (SEM 0.86) years and a mean BMI of 31.3 (SEM 0.8) kg/m2 completed the study. Subjects supplemented with whey protein had no significant change in body composition or serum glucose at 12 weeks compared with the control or casein group. Fasting TAG levels were significantly lowered in the whey group compared with the control group at 6 weeks (P = 0.025) and 12 weeks (P = 0.035). There was a significant decrease in total cholesterol and LDL cholesterol at week 12 in the whey group compared with the casein (P = 0.026 and 0.045, respectively) and control groups (P < 0.001 and 0.003, respectively). Fasting insulin levels and homeostasis model assessment of insulin resistance scores were also significantly decreased in the whey group compared with the control group (P = 0.049 and P = 0.034, respectively). The present study demonstrated that supplementation with whey proteins improves fasting lipids and insulin levels in overweight and obese individuals.


A Nanoprimer To Improve the Systemic Delivery of siRNA and mRNA.
Despite tremendous interest in gene therapies, the systemic delivery of nucleic acids still faces substantial challenges. To successfully administer nucleic acids, one approach is to encapsulate them in lipid nanoparticles (LNPs). However, LNPs administered intravenously substantially accumulate in the liver where they are taken up by the reticuloendothelial system (RES). Here, we administer prior to the LNPs a liposome designed to transiently occupy liver cells, the Nanoprimer. This study demonstrates that the pretreatment of mice with the Nanoprimer decreases the LNPs' uptake by the RES. By accumulating rapidly in the liver cells, the Nanoprimer improves the bioavailability of the LNPs encapsulating human erythropoietin (hEPO) mRNA or factor VII (FVII) siRNA, leading respectively to more hEPO production (by 32%) or FVII silencing (by 49%). The use of the Nanoprimer offers a new strategy to improve the systemic delivery of RNA-based therapeutics.


Reduced renal phosphate threshold concentration in alcoholic cirrhosis.
Hypophosphatemia is frequently found in patients with alcoholic cirrhosis, and its postulated causes include inadequate phosphorus (PO4) intake, reduced gastrointestinal absorption, and inappropriate PO4 loss in the urine. Of these, the first two would be expected to be associated with a high threshold concentration (Tm/GFR) and the last with a low Tm/GFR. 24 patients with alcoholic cirrhosis were studied. All had normal renal function. Simultaneous mid-morning serum and urine samples were obtained and analyzed for creatinine, Ca, PO4, and Mg. Median serum PO4 was 2.75 mg/dl, including 8 patients who were hypophosphatemic (serum PO4 less than 2.5 mg/dl). The median Tm/GFR was 2.3 mg/dl; 13 of the patients had values below 2.5 mg/dl. There was a significant correlation between serum PO4 and Tm/GFR (p less than 0.001). The median serum Ca was 8.5 mg/dl and did not correlate with Tm/GFR. The median serum Mg was 2.0 mg/dl. Serum magnesium was significantly correlated with Tm/GFR (p less than 0.05). We conclude that hypophosphatemia in alcoholic cirrhosis is associated with a low Tm/GFR and that this may be related to hypomagnesemia.


Fermentation, isolation, purification and characterization of an antitubercular antibiotic from Streptomyces luridus MTCC 4402.
Tuberculosis, caused by Mycobacterium tuberculosis, continues to be a serious public health problem around the world, and it urges the need for development of new antitubercular drugs. An antibiotic producing strain, Streptomyces luridus (MTCC 4402) was earlier isolated from soil by our group. In this work, the phylogenic status was established by 16S rRNA gene sequence analysis. The strain was found to be active against clinically resistant strains. The culture was grown in shake flasks in a medium containing cornsteep liquor, glucose, CaCO(3), soyabean meal and starch. Antibiotic production reached maximum at the end of 72 h. and fermentation profile was obtained. The active compound was extracted into ethyl acetate and was subjected to activity guided purification by column chromatography using silica gel, TLC and HPLC methods. The pure compound eluted at 16.7 min. by gradient elution was subjected to (1)H, (13)C NMR and mass spectral analyses. The acquired data was compared with that of natural products' data base and found to be a known antibiotic, spiramycin. The purified compound was studied for mutagenic, cytotoxicity, antitubercular activities. It was non mutagenic at the concentration of 1000 μg/mL, non cytotoxic and active as antitubercular agent at a concentration of 64 mg/mL and was comparable to rifampicin.


Impact of GLP-1 and GLP-1 receptor agonists on cardiovascular risk factors in type 2 diabetes.
Type 2 diabetes (T2D) is associated with increased cardiovascular disease and mortality. Most diabetes treatments have not proven to reduce this risk and may be associated with worsening of specific cardiovascular risk factors. GLP-1 receptor agonists (GLP-1R agonists) are new incretin-based therapies for the treatment of T2D. They improve glucose control by stimulating insulin secretion and suppressing glucagon release, both in a glucose-dependent manner. There are two GLP-1R agonists approved for the treatment of T2D: once daily liraglutide and twice daily exenatide, both administered by sc injection. Based on recent clinical trials, GLP-1R agonists suggest having a protective role in cardiovascular risk factors besides improving glycemic control, compared to placebo and to standard diabetes therapies. Both liraglutide and exenatide have demonstrated to induce clinically significant weight loss and to reduce systolic blood pressure. Liraglutide also has a positive effect on the lipid profile and cardiovascular risk biomakers. Furthermore, recent data shows a direct effect of GLP-1 and its metabolites in the vascular endothelium and the myocardium, leading to vasodilator effects and improved cardiac function in humans with acute myocardial infarction or congestive heart failure. GLP-1R agonists have a positive impact on cardiovascular risk factors otherwise not addressed by most standard diabetes therapies. Whether these new compounds actually decrease cardiovascular disease and mortality remains to be demonstrated in outcome studies.


Rare case of severe serotonin syndrome leading to bilateral compartment syndrome.
The term 'serotonin syndrome' describes a constellation of symptoms caused by serotonergic overstimulation. Its characteristic clinical presentation consists of encephalopathy, neuromuscular signs and autonomic hyperactivity. After removal of the offending agent, the clinical course is usually self-limited but can occasionally lead to severe symptoms. We report the case of a 68-year-old woman who presented emergently with encephalopathy. Home medications included paroxetine and dextroamphetamine/amphetamine. Physical examination revealed tachycardia, tachypnoea, diaphoresis, rigidity, hyperreflexia and clonus. Given the fast onset of symptoms, a diagnosis of serotonin syndrome was made. Laboratory studies showed acute-on-chronic kidney injury and elevated creatine kinase. The patient's mental status quickly returned to baseline with supportive care. Her rhabdomyolysis, however, persisted and led to acute compartment syndrome in her lower extremities. After bilateral leg fasciotomies and treatment of a severe wound infection with intravenous antibiotics, the patient has now recovered with complete resolution of her symptoms.


[The Chinese osteoporosis-targeted quality of life short questionnaire: design and reliability and validity assessment].
To design a disease-targeted life-of-quality questionnaire for Chinese patients with primary osteoporosis. From October 2013 to January 2014, 144 patients out- and inpatients were selected for assessment of their quality of life using the Chinese Osteoporosis-targeted Quality-of-Life short questionnaire (COQOL, 16 items) designed and reformed based on QUALEFFO-41. Statistics analysis was conducted to assess the validity of the COQOL. Statistics analysis showed good test-retest reliability, internal consistency, content validity, construct validity and criterion validity of COQOL. COQOL is reliable and valid for evaluating participants with low bone mass in clinical practice.


Dosimetric advantages of intensity modulated radiation therapy in locally advanced lung cancer.
Radiation therapy plays an essential role in the treatment of locally advanced lung cancer, but it inevitably leads to incidental and unnecessary dose to critical organs, including the lung, heart, and esophagus. Numerous radiation dose-volumetric parameters have been associated with increased risk of morbidity and mortality. The purpose of the present study is to quantify differences in normal tissue radiation exposure with intensity modulated radiation therapy (IMRT) compared with 3-dimensional conformal radiation therapy (3D-CRT). Twenty-four consecutive patients with locally advanced lung cancer undergoing definitive IMRT were enrolled on a phase 1 protocol. For each patient, an optimized 3D-CRT plan was also designed. Plans were normalized in terms of planning target coverage with a standard dose of 60 Gy in 2-Gy fractions with a subset of patients also receiving elective nodal irradiation to a dose of 44 Gy in 2-Gy fractions. Normal tissue dosimetric comparisons were made for the lung, heart, and esophagus. IMRT decreased incidental dose to the lungs, heart, and esophagus. For lung, both V20 Gy (21.5% vs 26.5%, P < .01) and mean lung dose (11.9 Gy vs 14.9 Gy, P < .01) were improved with IMRT without a corresponding increase in V5 Gy (P = .76). For heart, there was improvement in V5 (28.9% vs 33.7%, P < .01) but no difference in V30 Gy (9.8% vs 15.9%. P = .10). For esophagus, all dosimetric endpoints were improved (V20 Gy, V45 Gy, V60 Gy, mean dose). For example, V60 was 6.5% with IMRT compared with 21% with 3D-CRT (P < .01). IMRT significantly decreased unnecessary dose to critical organs with equivalent coverage of planning target volumes. IMRT may therefore improve the tolerability of therapy.


Orthodontically induced inflammatory root resorption. Part II: The clinical aspects.
Over the past 10 years, orthodontically induced inflammatory root resorption (OIIRR) has been increasingly recognized as an iatrogenic consequence of orthodontic treatment. With this in mind, orthodontists should take all known measures to reduce the occurrence of OIIRR. The evidence that we present in this review suggests several procedures known today that can avert this phenomenon; however, none of them can be relied on to completely prevent OIIRR. We believe that future studies might clarify the exact cause and course of OIIRR and, hopefully, help eliminate it. In Part I, we discussed the basic sciences aspects of OIIRR; in Part II, we present the clinical aspects of this phenomenon.


Patterns and management outcome of paediatrics burn in a general surgical department: experience from a secondary referral hospital.
Burns are considered the fourth most common type of traumatic injury worldwide with significant morbidity and mortality. The aim of this study is to disclose the incidence, etiological factors, treatment and outcome of paediatric burns in Sudan. This is a retrospective hospital-based study, conducted during 2016-2019. Patients who presented with burn injuries and managed in the hospital were included in the study. Flowchart sheets were used to collect the data. Then, it was entered and analysed using Statistical Package for the Social Sciences version 21 statistical software (IBM Corporation, Chicago, IL). The total number of patients who fulfill our criteria is 85 with a median age of 4 years (7 months-15 years). Sixty-five percent of them were under 5 years with male predominance (55%). Most patients' burns occurred at home. The median percentage of burn is 18% (2%-90%) which is caused by scalding burn among 55.3% followed by the flame burn in 41.2% of patients. Regarding fluid management, 60% received fluid. Open dressing for burn wounds was done for 75% and close dressing among 15% of patients. More than 80% of the patients in this study were treated and discharged in a good condition while the mortality was 5.9%. This study showed the significant burden of paediatric burns management on general surgery units in a referral hospital. Therefore, equipping the hospital and founding these specialities will decrease this load and will improve the outcome.


Who are the vulnerable lung cancer patients at risk for not receiving first-line curative or palliative treatment?
To identify non-small-cell lung cancer (NSCLC) patients in need of comprehensive support, we examined the association between patient and disease-related factors of vulnerability related to not receiving guideline-recommended treatment. We identified 14,597 non-small-cell lung cancer (NSCLC) patients with performance status <3 during 2013-2018 in the Danish Lung Cancer Registry. Multivariate logistic regression models were used to estimate Odds Ratios (ORs) and 95% confidence intervals (CIs) for receiving guideline-recommended treatment according to stage, comorbidities, age, performance status, long distance to hospital, cohabitation status, education and alcohol abuse. 21% of stage I-IIIA NSCLC patients did not receive curative treatment while 10% with stage IIIB-IV did not receive any oncological therapy. Factors associated with reduced likelihood of receiving curative treatment included: advanced stage (OR = 0.45; 95% CI = 0.42-0.49), somatic comorbidity (OR = 0.72; 95% CI = 0.63-0.83), age ≥ 80 years (OR = 0.59; 95% CI = 0.55-0.64), performance status = 2 (OR = 0.33; 95% CI = 0.28-0.39) and living alone (OR = 0.79; 95% CI = 0.69-0.90). Results were similar for stage IIIB-IV NSCLC patients, although a statistically significant association was also seen for long distances to the hospital (OR = 0.71; 95% CI = 0.58-0.86). Several factors are associated with not receiving guideline-recommended NSCLC treatment with age, performance status, comorbidity and stage being most predictive of no treatment receipt. Efforts should be made to develop support for vulnerable lung cancer patients to improve adherence to optimal first-line therapy.


Assessment of effective dose from concomitant exposures required in verification of the target volume in radiotherapy.
The requirement of the Ionising Radiation (Medical Exposure) Regulation 2000 [IR(ME)R] of justifying all exposures to ionizing radiation includes those from radiotherapy double exposure portal images resulting in exposure to normal tissues outside the treatment volume. Typical effective doses were calculated for a range of common sites using CT data to outline those parts of specific organs subject to concomitant radiation and generate dose-volume histograms. The product of the mean dose and the relative probability of inducing a fatal cancer in specific organs was used to determine a representative total effective dose in mSv per monitor unit for each site. A table of representative effective doses, ranging from 0.32 mSv to 2.56 mSv per monitor unit, was produced, which may be used to monitor cumulative effective doses of individual patients from double exposure portal images, in addition to those received from localization procedures.


Assessment of polychlorinated biphenyls and mercury levels in soil and biological samples from San Felipe, Nuevo Mercurio, Zacatecas, Mexico.
The aim of this work was to evaluate contamination by polychlorinated biphenyls and mercury in soil, and to assess exposure level to both contaminants in children living in San Felipe, Nuevo Mercurio, Zacatecas, Mexico. We found soil levels of total polychlorinated biphenyls ranging from non detectable (nd) to 190 μg/kg. Mercury soil levels ranged from 8.9 to 10215.0 mg/kg. Exposure levels of total polychlorinated biphenyls assessed in blood and urinary mercury in children living in the studied community were 1,600 ± 8,800 ng/g lipid and 4.2 ± 7.1 μg/g creatinine, respectively.


Testosterone and body functions.
Testosterone supplementation can help reduce many of the symptoms associated with androgen deficiency in the aging male by its effects on various parts of the body. Bone mineral density can decrease in the hypogonadal man and this may contribute to the increased fracture rate in the elderly. Testosterone therapy can improve bone mineral density and bone architecture by increasing bone formation and decreasing bone resorption - the possible benefits on fracture rate are unknown. Testosterone also improves body composition by reducing body fat mass and increasing lean body mass, and by increasing epidermal thickness, but its effects on muscle strength are still debated. In patients with diabetes and androgen deficiency, testosterone supplementation appears to reduce blood glucose and this could have important implications for cardiovascular risk reduction in patients with diabetes or the metabolic syndrome. The wide-ranging benefits of testosterone therapy in young and old men are clear and it appears that the route of administration (intramuscular, oral, or transdermal) does not alter this fact, but future work could illustrate even more profound effects of testosterone (e.g., in reducing cardiovascular risk) that could result in its recommended use in a wider range of patients.


Right hepatic artery injury associated with laparoscopic bile duct injury: incidence, mechanism, and consequences.
Because most bile duct injuries involve the common hepatic duct, the right hepatic artery, which is nearby, can also be injured. Reports on the frequency and significance of right hepatic artery injury (RHAI) associated with bile duct injury are sparse but suggest that RHAI increases mortality and decreases the success of the biliary repair. We studied the incidence, mechanism, and consequences of RHAI accompanying major bile duct injury. A total of 261 laparoscopic bile duct injuries were analyzed. Distribution was as follows: class I, 6%; class II, 22%; class III, 61%; and class IV, 11%. RHAI was present in 84 cases (32%): class I, 6%; class II, 17%; class III, 35% (P < 0.04 vs. class I/II); and class IV, 64% (P < 0.007 vs. class I/II/III). RHAI was more commonly associated with abscess, bleeding, hemobilia, right hepatic lobe ischemia, and subsequent hepatectomy (54% with RHAI vs. 11% without RHAI; P < 0.0001). RHAI had no influence on the success of the bile duct injury repair or on the mortality rate. Complications occurred more often with RHAI among cases repaired by the primary surgeon (41% RHAI vs. 2% no RHAI; P < 0.0001) but not among repairs by a biliary surgeon (3% RHAI vs. 2% no RHAI, P=NS; P < 0.0001 primary vs. biliary surgeon). RHAI increased morbidity, and occurred more often with class III and IV injuries reflecting the mechanisms of these injuries. RHAI did not increase the mortality rate or alter the success of biliary repair. Among biliary injuries repaired by the primary surgeon, RHAI was associated with a higher incidence of postoperative abscess, bleeding, hemobilia, hepatic ischemia, and the need for hepatic resection. A similar increase in the complication rate was not seen in patients treated by a biliary specialist.


A tale of two dogs: analyzing two models of canine ventricular electrophysiology.
The extensive development of detailed mathematical models of cardiac myocyte electrophysiology in recent years has led to a proliferation of models, including many that model the same animal species and specific region of the heart and thus would be expected to have similar properties. In this paper we review and compare two recently developed mathematical models of the electrophysiology of canine ventricular myocytes. To clarify their similarities and differences, we also present studies using them in a range of preparations from single cells to two-dimensional tissue. The models are compared with each other and with new and previously published experimental results in terms of a number of their properties, including action potential morphologies; transmembrane currents during normal heart rates and during alternans; alternans onsets, magnitudes, and cessations; and reentry dynamics of spiral waves. Action potential applets and spiral wave movies for the two canine ventricular models are available online as supplemental material. We find a number of differences between the models, including their rate dependence, alternans dynamics, and reentry stability, and a number of differences compared with experiments. Differences between models of the same species and region of the heart are not unique to these canine models. Similar differences can be found in the behavior of two models of human ventricular myocytes and of human atrial myocytes. We provide several possible explanations for the differences observed in models of the same species and region of the heart and discuss the implications for the applicability of models in addressing questions of mechanism in cardiac electrophysiology.


High follicle density does not decrease sweat gland density in Huacaya alpacas.
When exposed to high ambient temperatures, mammals lose heat evaporatively by either sweating from glands in the skin or by respiratory panting. Like other camelids, alpacas are thought to evaporate more water by sweating than panting, despite a thick fleece, unlike sheep which mostly pant in response to heat stress. Alpacas were brought to Australia to develop an alternative fibre industry to sheep wool. In Australia, alpacas can be exposed to ambient temperatures higher than in their native South America. As a young industry there is a great deal of variation in the quality and quantity of the fleece produced in the national flock. There is selection pressure towards animals with finer and denser fleeces. Because the fibre from secondary follicles is finer than that from primary follicles, selecting for finer fibres might alter the ratio of primary and secondary follicles. In turn the selection might alter sweat gland density because the sweat glands are associated with the primary follicle. Skin biopsy and fibre samples were obtained from the mid-section of 33 Huacaya alpacas and the skin sections were processed into horizontal sections at the sebaceous gland level. Total, primary, and secondary follicles and the number of sweat gland ducts were quantified. Fibre samples from each alpaca were further analysed for mean fibre diameter. The finer-fibred animals had a higher total follicle density (P<0.001) and more sweat glands (P<0.001) than the thicker-fibred animals. The fibre diameter and total follicle density were negatively correlated (R(2)=0.56, P<0.001). Given that the finer-fibred animals had higher follicle density and more sweat glands than animals with thicker fibres, we conclude that alpacas with high follicle density should not be limited for potential sweating ability.


Change of stretch reflex threshold in spasticity: effect of botulinum toxin injections.
Spasticity is a disorder of hypertonus associated with neurological diseases, characterized by a decrease in stretch reflex threshold. Stretch reflex threshold of wrist flexors has been recorded in subjects affected by forearm spasticity due to acute neurological lesions, occurred from one to sixty-one months before. In all the subjects a decreased stretch reflex threshold was recorded and a negative correlation between stretch reflex threshold and time of the disease resulted. In five subjects affected by mild spasticity the velocity stretch reflex threshold was tested one-three months after stroke and then six months later. In three cases a further decrease in stretch reflex threshold was recorded. Sixteen subjects affected by heavy forearm spasticity (quantified by Ashworth scale), were treated with Botulinum toxin injections to reduce spasticity. Fourteen of 16 subjects were responsive to the antispastic therapy: a decrease of at least 1 point in the Ashworth scale was detected after the treatment. In all the responsive cases an increase of stretch reflex threshold was recorded. The results confirm that the stretch reflex threshold is decreased in spastic muscles; it decreases progressively in time after the acute lesion. In addition, these results demonstrate that the decreased stretch reflex threshold can be reversed with Botulinum toxin injections. It is known that Botulinum toxin reduce the presynaptic release of Acetylcholine of neuromuscular synapses, but there are experimental evidences that it acts even on spindle's fibres, decreasing the sensitivity of intrafusal muscle fibres. This effect explains how Botulinum toxin increases the stretch reflex threshold in spastic muscles.


[Enhancement pattern of thyroid carcinoma with contrast-enhanced ultrasound].
To investigate the enhancement pattern of thyroid carcinoma with contrast-enhanced ultrasound (CEUS) for the diagnostic value. Thirty-one cases with thirty-five thyroid Occupied were retrospectively reviewed. The final diagnosis was confirmed by biopsy pathology. Contrast agent SonoVue and contrast pulse sequencing (CPS) technique were used in this study. Thirty-five thyroid carcinoma presented three enhancement patterns with CEUS. Type I: twenty-three lesions enhanced in a pattern of ring with centripetal fill-in, however, the central part of no contrast agent filling. Type II: five lesions enhanced regularly and homogeneously. Type III: seven lesions enhanced irregularly and homogeneously. Thyroid benign and malignant lesions perfusion time curve, compared to papillary thyroid carcinoma with nodular goiter in the AT, TTP and WT indicators P < 0.05; papillary thyroid carcinoma and thyroid adenoma, compared to WT alone an indicator of P < 0.05. Identification of the different enhancement patterns of thyroid carcinoma could improve the diagnostic ability of CEUS.


Metallothionein (I/II) suppresses genotoxicity caused by dimethylarsinic acid.
Arsenic is an environmental chemical of considerable concern due to its association with an increased risk of human cancer. Dimethylarsinic acid (DMAA) is one of the major methylated metabolites of ingested arsenicals in most mammals. To better clarify the role of metallothionein (MT) in modifying DMAA genotoxicity, MT-I/II null mice, and the corresponding wild-type mice, were exposed to DMAA (0, 188, 375 or 750 mg/kg body weight) via a single oral dose. Twenty-four hours after the DMAA injection, there was increased formation of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in serum and urine and a higher number of DNA strand breaks in peripheral blood cells. These increased levels were concomitant with increasing dose concentrations of DMAA in both strains of mice and they were significantly higher in MT-I/II null mice than in wild-type mice. Furthermore, the induction of apoptotic cells in the urinary bladder epithelium of MT-I/II null mice was significantly higher than in dose-matched wild-type mice exposed to DMAA. On the other hand, in both liver and in the alveolar and bronchial areas of the lung, the extent of DMAA-induced apoptosis was not different between wild-type and MT-I/II null mice and was increased in both strains. In addition, the concentration of hepatic MT in wild-type mice increased in a DMAA dose-dependent manner but was undetectable in MT-I/II null mice and could not subsequently be induced by DMAA. In conclusion, DMAA exposure causes oxidative stress, DNA damage and specific induction of apoptosis in target organs of arsenic carcinogenesis, which may be attributable to the mechanism(s) of arsenic-induced carcinogenesis in rodents. MT exhibited some protective roles during DNA damage presumably by acting as an antioxidant.


Physical Variables Underlying Tactile Stickiness During Fingerpad Detachment.
One may notice a relatively wide range of tactile sensations even when touching the same hard, flat surface in similar ways. Little is known about the reasons for this variability, so we decided to investigate how the perceptual intensity of light stickiness relates to the physical interaction between the skin and the surface. We conducted a psychophysical experiment in which nine participants actively pressed their finger on a flat glass plate with a normal force close to 1.5 N and detached it after a few seconds. A custom-designed apparatus recorded the contact force vector and the finger contact area during each interaction as well as pre- and post-trial finger moisture. After detaching their finger, participants judged the stickiness of the glass using a nine-point scale. We explored how sixteen physical variables derived from the recorded data correlate with each other and with the stickiness judgments of each participant. These analyses indicate that stickiness perception mainly depends on the pre-detachment pressing duration, the time taken for the finger to detach, and the impulse in the normal direction after the normal force changes sign; finger-surface adhesion seems to build with pressing time, causing a larger normal impulse during detachment and thus a more intense stickiness sensation. We additionally found a strong between-subjects correlation between maximum real contact area and peak pull-off force, as well as between finger moisture and impulse.


Spontaneous carbonate formation in an amorphous, amine-rich, polymeric drug substance: sevelamer HCl product quality.
Spectral differences among multiple manufacturers/lots of sevelamer HCl were observed by Fourier transform infrared spectroscopy, and further characterization was performed to identify the cause for these differences. The drug substance is a polymer that possesses a large molecular weight, is amorphous, and is practically insoluble in both water and organic solvents. Thus, solid-state characterization methods (spectroscopic and thermal) were required to identify and characterize differences among the samples to assess possible differences in product quality. ¹³C cross-polarization-magic-angle-spinning nuclear magnetic resonance spectroscopy of sevelamer HCl substances demonstrated the presence of a carbonyl-containing species, which was attributed to a carbonate impurity among samples. Stability studies demonstrated that this carbonate impurity formed spontaneously upon exposure of the drug substance to atmospheric water vapor and carbon dioxide, even under ambient conditions. Mechanistically, this behavior likely arises from the large number of primary and secondary amine groups, the hygroscopicity of the HCl salt, and a high degree of molecular mobility due to the amorphous nature of the drug substance.


Engineering a potent inhibitor of matriptase from the natural hepatocyte growth factor activator inhibitor type-1 (HAI-1) protein.
Dysregulated matriptase activity has been established as a key contributor to cancer progression through its activation of growth factors, including the hepatocyte growth factor (HGF). Despite its critical role and prevalence in many human cancers, limitations to developing an effective matriptase inhibitor include weak binding affinity, poor selectivity, and short circulating half-life. We applied rational and combinatorial approaches to engineer a potent inhibitor based on the hepatocyte growth factor activator inhibitor type-1 (HAI-1), a natural matriptase inhibitor. The first Kunitz domain (KD1) of HAI-1 has been well established as a minimal matriptase-binding and inhibition domain, whereas the second Kunitz domain (KD2) is inactive and involved in negative regulation. Here, we replaced the inactive KD2 domain of HAI-1 with an engineered chimeric variant of KD2/KD1 domains and fused the resulting construct to an antibody Fc domain to increase valency and circulating serum half-life. The final protein variant contains four stoichiometric binding sites that we showed were needed to effectively inhibit matriptase with a Ki of 70 ± 5 pm, an increase of 120-fold compared with the natural HAI-1 inhibitor, to our knowledge making it one of the most potent matriptase inhibitors identified to date. Furthermore, the engineered inhibitor demonstrates a protease selectivity profile similar to that of wildtype KD1 but distinct from that of HAI-1. It also inhibits activation of the natural pro-HGF substrate and matriptase expressed on cancer cells with at least an order of magnitude greater efficacy than KD1.


Sex-Dependent Association of Vitamin D With Insulin Resistance in Humans.
Animal studies suggested that vitamin D might decrease insulin resistance. Estrogen increased insulin sensitivity and glucose tolerance in rodents. However, sex-specific association of vitamin D with insulin resistance in humans remains unclear. To investigate the sex-dependency of the association of insulin resistance and 25-hydroxyvitamin D [25(OH)D] in a large Caucasian population. Cross-sectional study from out-patients' blood samples with measurements of 25(OH)D and homeostatic model assessment of insulin resistance (HOMA-IR) drawn at exactly the same day (n = 1887). This cohort was divided into 3 groups: (1) group with vitamin D deficiency (n = 1190), (2) group with vitamin D sufficiency (n = 686), and (3) vitamin D excess groups (n = 11); the vitamin D excess group was excluded from further analysis due to the small size. Analysis of the entire study population showed that serum 25(OH)D was inversely associated with HOMA-IR [Spearman correlation coefficient (rs) = -0.19, P < 0.0001]. When considering the vitamin D status, this association was only seen in the vitamin D deficiency group but not in the vitamin D sufficient group. The correlation was sex-dependent: HOMA-IR was inversely correlated with vitamin D in women with vitamin D deficiency (rs = -0.26, P < 0.0001) but not in men with vitamin D deficiency (rs = 0.01, P = 0.714). After multivariate linear regression analysis considering confounding factors, this relationship was again only seen in women. Vitamin D was inversely and independently associated with insulin resistance only in women with vitamin D deficiency. Based on our data, we suggest that in particular vitamin D deficient women might benefit from vitamin D substitution by improving insulin resistance. This, however, needs to be proven in adequately designed double-blind placebo-controlled clinical studies.


An unusual case of otogenic pneumocephalus.
Otogenic pneumocephalus is a rare entity usually caused by temporal bone trauma. This paper describes a case of otogenic pneumocephalus of traumatic origin, in which the type of the fracture (a bony spicula was detached from the mastoid) and the location (Trautmann's triangle) were uncommon.