[Evaluation of CT, MRI and PET-CT in detecting retropharyngeal lymph node metastasis in nasopharyngeal carcinoma].
At present, the detection of retropharyngeal lymph node (RLN) metastasis in nasopharyngeal carcinoma (NPC) mainly depends on imaging, but the diagnosis criteria are controversial. Recently, the 18F-fluoro-2-deoxy-D-glucose positron emission tomography ((18)FDG-PET-CT) has been widely applied in diagnosing cancer. This study was to evaluate CT, MRI, and (18)FDG-PET-CT in detecting RLN metastasis of NPC, and explore the imaging diagnosis criteria. From Dec. 2003 to Aug. 2005, 53 consecutive patients with NPC treated in Cancer Center of Sun Yat-sen University were enrolled. Contrast enhanced CT, MRI, and (18)FDG-PET-CT were performed on bilateral RLNs of the patients within 10 days before treatment. The diagnosis criteria of RLN metastasis for CT and MRI included single node with maximum diameter of > or =4 mm, and the presence of central necrosis or ringed enhancement; the criteria for (18)FDG-PET-CT included abnormal accumulation of irradiation in the RLN with the standard uptake value (SUV) of > or =2.5. Considering the response of RLN to clinical treatment, the differences of CT, MRI, and PET-CT in detecting RLN metastasis were compared. Among the 106 intropharyngeal spaces of the 53 patients, the positive rate of RLN metastasis was significantly higher detected by CT and MRI than by PET-CT (39.6% and 45.3% vs. 20.8%, P<0.01), but the difference between CT and MRI was not significant (P>0.05). CT detected 27 cases of RLN metastasis with the maximum diameter of 0.4-0.9 cm, MRI detected 31 cases, and PET-CT only detected 5 cases. Especially, MRI detected 7 cases of RLN metastasis with the maximum diameter of < or =0.5 cm, CT detected 4 cases, but PET-CT detected none. CT detected 12 cases of RLN metastasis with the maximum diameter of > or =1.0 cm, MRI detected 14 cases, and PET-CT detected 13 cases. All patients were followed-up after treatment. One case of RLN metastasis with necrosis and the maximum diameter of 1.0 cm, and 1 case with the maximum diameter of 1.5 cm were eliminated within 6 months after treatment; the rest were eliminated within 3 months. In detecting RLN metastasis in NPC, MRI has no obvious advantage over CT, while PET-CT is markedly inferior to CT and MRI, especially in detecting the cases with the maximum diameter of <1.0 cm. Furthermore, the metastatic RLNs with necrosis or the maximum diameter of < or =5 mm can't be detected by PET-CT. Setting the maximum diameter of > or =4 mm as a diagnosis criterion of RLN metastasis for CT or MRI is reasonable and feasible.


The complete mitochondrial genome of Matuta victor (Decapoda: Matutidae) from Beibu Bay.
The true crabs (Brachyura) including Calappidea are one of the most diverse groups of Decapod crustaceans However, despite their great diversity and commercial importance, phylogenetic and classification relationships within Calappidea are still complicated and controversial. In this study, we report the first complete mitochondrial genome of Matuta victos. The mitogenome has 17,782 base pairs (70.1% A + T content) and is made up of a total of 37 genes (13 protein-coding, 22 transfer RNAs and two ribosomal RNAs), plus a putative control region. This study will provide useful molecular resources for clarifying evolutionary and phylogenetic confusion within Calappidea.


The pediatrics milestones: conceptual framework, guiding principles, and approach to development.
The Accreditation Council for Graduate Medical Education (ACGME) and the American Board of Pediatrics (ABP) have partnered to initiate the Pediatrics Milestone Project to further refine the 6 ACGME competencies and to set performance standards as part of the continued commitment to document outcomes of training and program effectiveness. Members of the Pediatrics Milestone Project Working Group searched the medical literature and beyond to create a synopsis of models and evidence for a developmental ontogeny of the elements for 52 subcompetencies. For each subcompetency, we created a series of Milestones, grounded in the literature. The milestones were vetted with the entire working group, engaging in an iterative process of revisions until reaching consensus that their narrative descriptions (1) included all critical elements, (2) were behaviorally based, (3) were properly sequenced, and (4) represented the educational continuum of training and practice. We have completed the first iteration of milestones for all subcompetencies. For each milestone, a synopsis of relevant literature provides background, references, and a conceptual framework. These milestones provide narrative descriptions of behaviors that represent the ontogeny of knowledge, skill, and attitude development across the educational continuum of training and practice. The pediatrics milestones take us a step closer to meaningful outcome assessment. Next steps include undertaking rigorous study, making appropriate modifications, and setting performance standards. Our aim is to assist program directors in making more reliable and valid judgments as to whether a resident is a "good doctor" and to provide outcome evidence regarding the program's success in developing doctors.


Within-person variability in curiosity during daily life and associations with well-being.
Curiosity promotes engagement in novel situations and the accruement of resources that promote well-being. An open question is the extent to which curiosity lability, the degree to which curiosity fluctuates over short timescales, impacts well-being. We use data from a 21-day daily diary as well as trait measures in 167 participants (mean age = 25.37 years, SD = 7.34) to test (a) the importance of curiosity lability for depression, flourishing, and life satisfaction, (b) day-to-day associations among curiosity and happiness, depressed mood, anxiety, and physical activity, and (c) the role of day's mood as a mediator between physical activity and curiosity. We observe positive associations among curiosity lability and depression, as well as negative associations among curiosity lability and both life satisfaction and flourishing. Curiosity is higher on days of greater happiness and physical activity, and lower on days of greater depressed mood. We find evidence consistent with day's depressed mood and happiness being mediators between physical activity and curiosity. Greater consistency in curiosity is associated with well-being. We identify several potential sources of augmentation and blunting of curiosity in daily life and provide support for purported mechanisms linking physical activity to curiosity via mood.


Radiological diagnosis of developmental disorders of the nervous system and adjacent structures.
On the basis of the author's own experiences and drawing on data available in the literature, present possibilities of neuroradiological diagnosis of developmental disorders of the spine and skull, developmental anomalies of meninges of the brain and the spinal cord and of the central nervous system, the brain and the spinal cord are discussed. The rapid development of investigative methods in the past decades has brought about an immense amount of new information gained in the diagnosis of these anomalies and has facilitated their early detection and a more accurate evaluation. The morphological aspect is not, however, the only factor in the estimation of the clinical relevance of these disorders. For this reason not only the morphological, but also the functional relationship of developmental disorders of the nervous system to the adjacent structures is emphasized and, on the other hand, the influence of these anomalies on the nervous system is discussed. Many years' experiences in clinical neurology and problems of manual medicine indicate that these relations may be the cause of the various clinical symptoms. In this respect the presented findings mean a contribution to the treatment of the clinical complaints that are pathogenetically related to developmental disorders.


Compatibility between itinerant synaptic receptors and stable postsynaptic structure.
The density of synaptic receptors in front of presynaptic release sites is stabilized in the presence of scaffold proteins, but the receptors and scaffold molecules have local exchanges with characteristic times shorter than that of the receptor-scaffold assembly. We propose a mesoscopic model to account for the regulation of the local density of receptors as quasiequilibrium. It is based on two zones (synaptic and extrasynaptic) and multilayer (membrane, submembrane, and cytoplasmic) topological organization. The model includes the balance of chemical potentials associated with the receptor and scaffold protein concentrations in the various compartments. The model shows highly cooperative behavior including a "phase change" resulting in the formation of well-defined postsynaptic domains. This study provides theoretical tools to approach the complex issue of synaptic stability at the synapse, where receptors are transiently trapped yet rapidly diffuse laterally on the plasma membrane.


Diagnosis of hantavirus infection in humans and rodents in Ribeirão Preto, State of São Paulo, Brazil.
Hantavirus pulmonary and cardiovascular syndrome (HPCS) is an emerging serious disease in the Americas. Hantaviruses (Bunyaviridae) are the causative agents of this syndrome and are mainly transmitted through inhalation of aerosols containing the excreta of wild rodents. In the Ribeirão Preto region (state of São Paulo, Brazil), HPCS has been reported since 1998, caused by the Araraquara virus (ARAV), for which Necromys lasiurus is the rodent reservoir. This study aimed to show diagnostic results relating to infection in humans and rodents, obtained at the Virology Research Center of the Ribeirão Preto School of Medicine, University of São Paulo, between 2005 and 2008. HPCS was diagnosed by means of ELISA and/or RT-PCR in 11 (21.2%) out of 52 suspected cases, and 54.4% of these were fatal. Furthermore, 595 wild rodents (Necromys lasiurus, Akodon sp, Calomys tener and Oligoryzomys sp) were caught between 2005 and 2008. Fifteen (2.5%) of these rodents presented antibodies for hantavirus, as follows: Necromys lasiurus (4%), Calomys tener (1.9%) and Akodon sp (1.5%). Nucleotide sequences obtained through RT-PCR from one HPCS patient and one Calomys tener rodent were compared with hantavirus sequences from GenBank, which showed that both were homologous with ARAV. This work corroborates previous studies showing that ARAV is the hantavirus causing HPCS in the Ribeirão Preto region. It also shows that rodents infected with hantavirus represent a constant risk of transmission of this virus to man.


Tetranitromethane oxidation of phytochrome chromophore as a function of spectral form and molecular weight.
Tetranitromethane bleaches Avena phytochrome. The phytochrome (far-red absorbing form; Pfr) chromophore of 124 kilodalton (kD) phytochrome is oxidized 8 times more rapidly than the red absorbing form (Pr). Proteolysis of the 124 kD molecule to the extensively studied mixture of 118 and 114 kD polypeptides increases the rate of oxidation of Pfr 5-fold without affecting the rate of Pr oxidation. As a result, the Pfr form of 118/114 kD preparations is oxidized at a rate 40 times greater than the Pr form. Further proteolytic degradation of the chromoprotein to 60 kD results in an additional increase in the oxidation rates of both Pr and Pfr. These differences in reactivity to tetranitromethane indicate that the chromophore of Pfr is either intrinsically more chemically reactive and/or physically more accessible than the Pr chromophore and that the reactivity/accessibility of both spectral forms is increased by proteolysis. The enhanced reactivity of the Pfr chromophore after proteolytic cleavage of the 6 to 10 kD polypeptide segment(s) from the 124 kD species is further evidence that these segment(s) affect the environment of the native photoreceptor.


New optical supramolecular compound constructed from a polyoxometalate cluster and an organic substrate.
A new optical supramolecular compound constructed from a polyoxometalate cluster and an organic substrate [(H3O)(C12H10N3)2(PW12O40)] (1) has been synthesized via a hydrothermal reaction and has been structurally characterized by X-ray diffraction. The solid-state diffuse reflectance, IR, and photoluminescence spectra of the title compound indicate that there is an interaction between the alpha-PW12O40 and the organic substrate. The light-yellow title compound shows a certain second-order nonlinear optical response of I(2omega) = 2I(KDP)(2omega).


QTL mapping for genetic determinants of lipoprotein cholesterol levels in combined crosses of inbred mouse strains.
To identify additional loci that influence lipoprotein cholesterol levels, we performed quantitative trait locus (QTL) mapping in offspring of PERA/EiJxI/LnJ and PERA/EiJxDBA/2J intercrosses and in a combined data set from both crosses after 8 weeks of consumption of a high fat-diet. Most QTLs identified were concordant with homologous chromosomal regions that were associated with lipoprotein levels in human studies. We detected significant new loci for HDL cholesterol levels on chromosome (Chr) 5 (Hdlq34) and for non-HDL cholesterol levels on Chrs 15 (Nhdlq9) and 16 (Nhdlq10). In addition, the analysis of combined data sets identified a QTL for HDL cholesterol on Chr 17 that was shared between both crosses; lower HDL cholesterol levels were conferred by strain PERA. This QTL colocalized with a shared QTL for cholesterol gallstone formation detected in the same crosses. Haplotype analysis narrowed this QTL, and sequencing of the candidate genes Abcg5 and Abcg8 confirmed shared alleles in strains I/LnJ and DBA/2J that differed from the alleles in strain PERA/EiJ. In conclusion, our analysis furthers the knowledge of genetic determinants of lipoprotein cholesterol levels in inbred mice and substantiates the hypothesis that polymorphisms of Abcg5/Abcg8 contribute to individual variation in both plasma HDL cholesterol levels and susceptibility to cholesterol gallstone formation.


Angiotensin II Receptor Physiology Using Gene Targeting.
The study of mice with targeted disruptions of angiotensin receptor genes has provided new insights into the roles of the individual receptor subtypes, i.e., AT(1A), AT(1B), and AT(2), in growth, development, and the regulation of blood pressure.


Subconjunctival Loa Loa worm: case report.
A 38-year old Ghanaian suddenly had the sensation of a foreign body in his right eye. Slit-lamp examination revealed a transparent worm underneath the conjunctiva. With topical anesthesia, the complete worm, 3.5 cm long, was removed surgically. A microbiological analysis at the Institute of Tropical Medicine confirmed the diagnosis of Loa Loa. Laboratory tests showed negative blood eosinophilia, positive blood film examination for microfilariae and positive results for filarial serology. The postoperative treatment consisted of progressive doses of di-ethyl-carbamazine (50-->100-->200 mg/d). A subconjunctival Loa Loa worm is rare in Belgium and usually occurs in immigrants or travellers returning from Tropical (Equatorial) West and Central Africa. Our patient visited Nigeria in 1985 and Ivory Coast in 1986. Those regions are highly endemic for Loa Loa.


Risk factors for candidaemia and their cumulative effect over time in a cohort of critically ill, non-neutropenic patients.
There is an increasing incidence of invasive candidal infections in critically ill patients worldwide, which has prompted development of various risk prediction rules, both clinical and microbiological. To date, however, there is a lack of research into how cumulative risk factors over time affect transition to candidaemia. The aim of this study was to investigate the association of risk factor accumulation over time with candidaemia in a cohort of critically ill, non-neutropenic adult patients. A single centre, retrospective, matched case-control study in a tertiary referral intensive care unit (ICU). Data were retrieved and analysed from 108 patients (54 cases and 54 controls) admitted between 1 January 2008 and 1 August 2016. Primary outcome was the association between time-dependent risk factors and candidaemia. Secondary outcomes were ICU and inhospital mortality. Baseline demographic and clinical factors were similar across both groups. Time dependent univariate factors associated with candidaemia were days of mechanical ventilation, systemic antibiotic use, renal replacement therapy, central venous access, total parenteral nutrition (TPN), systemic inflammatory response syndrome, Candida site colonisation and number of surgeries. Factors persisting on multivariate analysis were days of TPN use (odds ratio [OR], 1.8; 95% CI, 1.02-3.22; P = 0.041) and total Candida site colonisation days (OR, 2.41; 95% CI, 1.30-4.46; P = 0.005). Mortality and length of stay (LOS) was greater in patients with candidaemia v control patients (ICU mortality, 15 [28%] v 10 [19%]; P = 0.254; hospital mortality, 26 [48%] v 16 [30%]; P = 0.048; ICU LOS median, 13 days [interquartile range (IQR), 5-29 days] v 2 days [IQR, 1-5 days]; P < 0.001; hospital LOS median, 36 days [IQR,19- 63 days] v 13 days [IQR, 6-28 days]; P < 0.001). This study demonstrates an association between TPN use, Candida colonisation and cumulative risk over time of developing candidaemia.


Analysis of lorazepam in rat brain using liquid/liquid and solid-phase extraction in combination with high performance liquid chromatography.
A method for the determination of lorazepam in rat brain is described using liquid/liquid and solid-phase extraction, followed by high performance liquid chromatography. After addition of chlordiazepoxide as the internal standard, 100 mg brain tissue was homogenized and incubated with alkaline protease. Lorazepam and chlordiazepoxide were extracted three times with toluene. After treatment through a C18-Bond Elut column, lorazepam and chlordiazepoxide were analyzed isocratically on a reversed-phase column with a mobile phase consisting of methanol +0.025 M sodium phosphate buffer (66:34, v/v). The eluted drugs were monitored by their absorption at 240 nm. The sensitivity limit of this method was 10 ng of lorazepam per 100 mg of brain tissue sample. The standard curve was linear over the range of 20 to 200 ng lorazepam. The coefficient of variation for day-to-day precision established by 21 replicate analyses was 4.5 to 13.6%.


[The importance of water testing for public health in two regions in Rio de Janeiro: a focus on fecal coliforms, nitrates, and aluminum].
In developing countries, due to poor sanitation conditions and poor quality of drinking water, typical water-borne diseases and more recently diseases caused by drinking water with high concentrations of nitrates and certain metals like aluminum have increased the concern over the health effects of these compounds. Several articles have shown associations between nitrates and methemoglobinemia in children, and aluminum and Alzheimer disease in adults. This study identified water quality with several parameters in non-conformity with Brazilian drinking water standards (Ruling 36/90): more than 50% of all samples from both regions contained fecal coliforms; some 31% of water samples from wells in Duque de Caxias had excessive nitrate concentrations; 100% of all groundwater samples from both regions showed aluminum concentrations not conforming to the norm, with the same result for 100% of samples from the drinking water distribution system in São Gonçalo and 75% of same in Duque de Caxias. This lack of conformity poses several health risks for the local population.


FloTrac/Vigileo system monitoring in acute-care surgery: current and future trends.
As acute critical-care surgery evolves, it is imperative to introduce reliable devices that can intraoperatively assess a patient's cardiovascular functions. Owing to the fact that traditional methods are usually invasive, non- or less-invasive innovations have attracted the attention of clinicians in recent decades. The FloTrac system monitors cardiovascular performance by analyzing peripheral arterial waveforms and a preset database, and it decreases the invasiveness by using a pulmonary arterial catheter. The reliability of cardiac output measurements was confirmed in many critically ill subjects in cardiac surgeries and intensive care units. Moreover, the FloTrac system is easy to set up, and interpreting the information is simple. The FloTrac system also provides a useful preload predictor, that is, stroke volume variation (SVV), for fluid management, which has been proven to enhance surgical safety in the treatment of critically ill patients. Goal-directed therapy guided by SVV and other hemodynamic variables was advocated for peri-operative fluid optimization. Although the evolution of each updated algorithm of the FloTrac system has demonstrated improved accuracy and limited shortcomings, the latest third-generation algorithm is still not equal to the gold standard reference. The accuracy of the latest third-generation algorithm is controversial in septic conditions, and its use is still unacceptable in liver transplantation. Due to vasoactive challenges, especially in the administration of norepinephrine, a conclusion could not be reached. Clinicians should recognize the appropriate uses and limitations when using the algorithm during acute critical surgeries.


Posttranscriptional regulation of interferon mRNA levels in peritoneal macrophages.
Low levels of beta interferon (IFN) mRNA are transcribed in freshly explanted murine peritoneal macrophages. Nuclear runoff transcription assays show that this "constitutive" IFN-beta-mRNA transcription does not increase in macrophages treated either with lipopolysaccharide or with IFN-gamma, which induce a marked accumulation of this mRNA and greatly increase IFN secretion. Therefore, these agents promote accumulation of IFN-beta mRNA by posttranscriptional mechanisms. The IFN-alpha 2 gene is also constitutively transcribed by macrophages, but the corresponding mRNA does not accumulate in lipopolysaccharide-treated cells.


Nur77 prevents excessive osteoclastogenesis by inducing ubiquitin ligase Cbl-b to mediate NFATc1 self-limitation.
Osteoclasts are bone-resorbing cells essential for skeletal remodeling. However, over-active osteoclasts can cause bone-degenerative disorders. Therefore, the level of NFATc1, the master transcription factor of osteoclast, must be tightly controlled. Although the activation and amplification of NFATc1 have been extensively studied, how NFATc1 signaling is eventually resolved is unclear. Here, we uncover a novel and critical role of the orphan nuclear receptor Nur77 in mediating an NFATc1 self-limiting regulatory loop to prevent excessive osteoclastogenesis. Nur77 deletion leads to low bone mass owing to augmented osteoclast differentiation and bone resorption. Mechanistically, NFATc1 induces Nur77 expression at late stage of osteoclast differentiation; in turn, Nur77 transcriptionally up-regulates E3 ubiquitin ligase Cbl-b, which triggers NFATc1 protein degradation. These findings not only identify Nur77 as a key player in osteoprotection and a new therapeutic target for bone diseases, but also elucidate a previously unrecognized NFATc1→Nur77→Cblb-•NFATc1 feedback mechanism that confers NFATc1 signaling autoresolution.


Mycobacterium tuberculosis of the Beijing Genotype in Iran and the World Health Organization Eastern Mediterranean Region: A Meta-Analysis.
The Beijing genotype is a distinct genetic lineage of Mycobacterium tuberculosis, which is distributed worldwide, and may cause large outbreaks of multidrug resistance-tuberculosis (MDR-TB). The distribution of such strains in the Eastern Mediterranean region (EMR) is unclear, and clarifying the data is our purpose apart from the presence of Beijing TB in Iran. We searched Published literature from CINAHL Cochrane Library, Current Contents, Database of Abstracts of Reviews of Effects (DARE), ProQuest Google Scholar PubMed, PsycINFO, Thomson Reuters, (SID), and Medical Library (MedLib) to detect relevant studies from the year 2000 to July 2015 with the following keywords: M. tuberculosis, Beijing genotype, EMR, and drug resistance. Random-effect models were used to estimate the proportion of Beijing strains in STATA 14. Heterogeneity was investigated by subgroup analysis and meta-regression. The meta-prevalence of Beijing strains was 4% (CI 95% = 3-5). The prevalence was different based on types of detection techniques (spoligotyping = 4% vs. other techniques = 6%; p = 0.003) and years of study (before the year 2000 = 2% vs. after year 2000 = 4%, p = 0.004). The Beijing family was most prevalent in Iran and Pakistan. A strong relationship with drug resistance was reported in Pakistan and Iran, and an increasing trend was seen in Pakistan. Additional studies of drug-resistant TB distribution among Beijing strains in EMR countries are needed as well as a time-trend analysis of the Beijing strain infection in the region.


[Diagnosis of viral hepatitis in 1991].
Viral hepatitis is a disease which is frequent but often asymptomatic and therefore remains unrecognized until a late stage. Hence the importance of biochemical and, chiefly, virological detection in populations at risk such as transfusion recipients, drug addicts or exposed professionals. Detection is based on systematic transaminase assays during check-ups or in the presence of warning signs. Any rise in transaminase level must lead to serological examinations in search of the A, B, C and Delta markers. Routine vaccination of subjects at risk against hepatitis B should considerably reduce the prevalence of this disease and of Delta hepatitis in the populations exposed.


Somatosensory evoked potentials induced by stimulating a variable number of nerve fibers in rat.
Somatosensory evoked potentials (SEPs) were recorded from rat spinal cord (sSEPs) and cerebral cortex (cSEPs). Stimulus sites included either one or both sural nerve branches having different fiber populations (group A), or distal to a lesion of controlled size of the sural nerve made 1 week earlier (group B). In the two groups of animals, amplitudes of SEPs correlated with the quantity of large myelinated nerve fibers. Peak latencies of sSEPs in group A related to the ratio of sizes of transmitting fibers. sSEPs and cSEPs in both groups A and B could be recorded in a reproducible fashion by stimulating sural nerve branches or lesioned nerve trunks containing only 100 or less nerve fibers greater than 4 microns in size. Thus, presence of sSEPs or cSEPs after stimulation distal to a lesion site does not insure that many nerve fibers have continuity with the central nervous system (CNS).


TGF-beta IL-6 axis mediates selective and adaptive mechanisms of resistance to molecular targeted therapy in lung cancer.
The epidermal growth-factor receptor (EGFR) tyrosine kinase inhibitor erlotinib has been proven to be highly effective in the treatment of nonsmall cell lung cancer (NSCLC) harboring oncogenic EGFR mutations. The majority of patients, however, will eventually develop resistance and succumb to the disease. Recent studies have identified secondary mutations in the EGFR (EGFR T790M) and amplification of the N-Methyl-N'-nitro-N-nitroso-guanidine (MNNG) HOS transforming gene (MET) oncogene as two principal mechanisms of acquired resistance. Although they can account for approximately 50% of acquired resistance cases together, in the remaining 50%, the mechanism remains unknown. In NSCLC-derived cell lines and early-stage tumors before erlotinib treatment, we have uncovered the existence of a subpopulation of cells that are intrinsically resistant to erlotinib and display features suggestive of epithelial-to-mesenchymal transition (EMT). We showed that activation of TGF-beta-mediated signaling was sufficient to induce these phenotypes. In particular, we determined that an increased TGF-beta-dependent IL-6 secretion unleashed previously addicted lung tumor cells from their EGFR dependency. Because IL-6 and TGF-beta are prominently produced during inflammatory response, we used a mouse model system to determine whether inflammation might impair erlotinib sensitivity. Indeed, induction of inflammation not only stimulated IL-6 secretion but was sufficient to decrease the tumor response to erlotinib. Our data, thus, argue that both tumor cell-autonomous mechanisms and/or activation of the tumor microenvironment could contribute to primary and acquired erlotinib resistance, and as such, treatments based on EGFR inhibition may not be sufficient for the effective treatment of lung-cancer patients harboring mutant EGFR.


Non-small-cell lung carcinoma: role of the Notch signaling pathway.
Notch signaling plays a pivotal role during embryogenesis. It regulates three fundamental processes: lateral inhibition, boundary formation, and lineage specification. During post-natal life, Notch receptors and ligands control critical cell fate decisions both in compartments that are undergoing differentiation and in pluripotent progenitor cells. First recognized as a potent oncogene in certain lymphoblastic leukemias and mesothelium-derived tissue, the role of Notch signaling in epithelial, solid tumors has been far more controversial. The overall consequence of Notch signaling and which form of the Notch receptor drives malignancy in humans is deeply debated. Most likely, this is due to the high degree of context-dependent effects of Notch signaling. More recently, it has been discovered that Notch (especially Notch-1) can exert different, even opposite effects in the same tissue under differing microenvironmental conditions. Further complicating the understanding of Notch receptors is the recently discovered role for non-canonical Notch signaling. Additionally, the most frequent Notch signaling antagonists used in biological systems have been inhibitors of the transmembrane protease complex γ-secretase, which itself processes a plethora of class one transmembrane proteins and thus cannot be considered a Notch-specific upstream regulator. Here we review the available empirical evidence gathered in recent years concerning Notch receptors and ligands in non-small-cell lung carcinoma (NSCLC). Although an overview of the field reveals seemingly contradicting results, we propose that Notch signaling can be exploited as a therapeutic target in NSCLC and represents a promising complement to the current arsenal utilized to combat this malignancy, particularly in targeting NSCLC tissues under specific environmental conditions, such as hypoxia.


Low-frequency ultrasound induces nonenzymatic thrombolysis in vitro.
To evaluate whether ultrasound, applied over a distance of several centimeters and in the absence of thrombolytic agents, may have a thrombolytic effect on blood clots. Low-frequency (20 kHz) continuous wave ultrasound at different intensity levels (0.15-1.2 W/cm2) and exposure times (5, 10, and 20 minutes) was assessed for its potential to induce thrombolysis of fresh human blood clots. The ultrasound effect was also studied in combination with recombinant tissue-type plasminogen activator-mediated thrombolysis. Experiments were carried out in a flow model in degassed sodium phosphate buffer at 37 degrees C at a distance of 3 cm from the ultrasonic probe to the blood clots. Regardless of ultrasound exposure times, blood clots in all experimental groups and the control group were left in the flow system for 20 minutes. The use of ultrasound alone showed a significant thrombolytic effect compared with the control group, with a statistically significant effect at 0.15 W/cm2 and exposure of 10 minutes (P = .02). There was a clear correlation between the extent of weight loss and the chosen intensity level and exposure time. Complete disruption in 8 of 10 blood clots occurred at 1.2 W/cm2 within 10 min. Addition of ultrasound to recombinant tissue-type plasminogen activator-mediated thrombolysis significantly enhanced thrombolysis compared with application of recombinant tissue-type plasminogen activator or ultrasound alone (P = .0001), with the results pointing toward a purely additive, nonsynergistic effect of the 2 treatment modalities. Lysis was more effective in fresh thrombi. The use of low-frequency ultrasound alone, without addition of a thrombolytic drug, has the potential to induce thrombolysis over a distance. Combination of ultrasound with recombinant tissue-type plasminogen activator is superior to either treatment alone. Ultrasound is a promising tool for developing an alternative or additional treatment modality for acute cerebral vessel occlusion.


Insulin enhances endothelial alpha2-adrenergic vasorelaxation by a pertussis toxin mechanism.
To investigate whether insulin effect on endothelium is related to a specific signal transduction pathway or reflects a more generalized action of the hormone, we studied in aortic rings of Wistar-Kyoto (WKY) rats the effects of the hormone on endothelium-dependent relaxations generated by acetylcholine, adenosine diphosphate, the selective alpha2-adrenergic agonist UK 14,304, and the calcium ionophore ionomycin. The responses were evaluated both in control conditions and after 30 minutes of exposure to three different levels of insulin (30, 100, and 500 microU/mL). Insulin failed to modify the phenylephrine aortic contractions and the relaxations induced by acetylcholine, adenosine diphosphate, and ionomycin. In contrast, both 100 and 500 microU/mL insulin were able to potentiate the UK 14,304-induced vasorelaxation (+96+/-19% and +91+/-12%, respectively). Pertussis toxin, which causes alpha2-adrenergic receptor Gi uncoupling, reduced the alpha2-adrenergic vasorelaxation and prevented the insulin potentiation of the response to UK 14,304. Furthermore, in primary cultured aortic endothelial cells from WKY, we evaluated the conversion of [3H]arginine to [3H]citrulline in response to acetylcholine, ionomycin, and UK 14,304, both in control conditions and during insulin exposure. Again, insulin did not affect basal citrulline production or the increase induced by acetylcholine and ionomycin, whereas it potentiated the response to UK 14,304. Finally, in aortic rings of spontaneously hypertensive rats, insulin treatment (100 and 500 microU/mL) was unable to enhance the alpha2-adrenergic vasodilator response; in vascular endothelial cells from spontaneously hypertensive rats, insulin did not potentiate the increase in citrulline production evoked by UK 14,304. In conclusion, insulin selectively enhances alpha2-adrenergic endothelial vasorelaxation through a pertussis toxin-sensitive mechanism, by potentiating endothelial nitric oxide production. This vasorelaxant mechanism is altered in spontaneously hypertensive rats.


Human Histologic Evidence of Reosseointegration Around an Implant Affected with Peri-implantitis Following Decontamination with Sterile Saline and Antiseptics: A Case History Report.
The treatment of peri-implant disease is one of the most controversial topics in implant dentistry. The multifactorial etiology and the myriad proposed techniques for managing the problem make successful decontamination of an implant surface affected by peri-implantitis one of the more unpredictable challenges dental practitioners have to face. This article presents the first known published case report demonstrating human histologic evidence of reosseointegration using a plastic curette for mechanical debridement and dilute sodium hypochlorite, hydrogen peroxide, and sterile saline for chemical detoxification. Guided bone regeneration in the infrabony component of the peri-implantitis lesion was accomplished using calcium sulfate and bovine bone as grafting materials and a porcine collagen barrier for connective tissue and epithelial exclusion.


The value of positron-emission tomography/computed tomography for radiotherapy treatment planning: a single institutional series.
This study aimed to compare the clinical target volumes (CTV) delineated by computed tomography (CT) and positron emission tomography (PET)/CT using fluoro-deoxy-glucose to assess the impact of using PET information for radiotherapy (RT) planning. We retrospectively reviewed data on patients with tumors from different sites who had indications for RT and had undergone RT treatment planning with PET/CT at our institution between July 2003 and July 2009. Statistical analysis included the comparison of CTV planned for treatment only by CT (CTV(CT)) with that by PET/CT (CTV(PET)) using the Wilcoxon test for paired samples. Of 105 patients eligible for analysis, 56.2% were men. The most common diagnoses were head and neck cancer (28.6%), lung cancer (21.9%), lymphoma (11.4%), upper gastrointestinal (10.5%), and others (14.3%). Comparison of CTV(CT) and CTV(PET) revealed that RT planning has changed in 77% of cases because of PET/CT additional information, with impact on treatment volume varying according to diagnosis. Despite the absolute difference observed between median CTVs, there was no significant difference between CTV(CT) and CTV(PET) (114 vs. 90.4 ml, respectively; P=0.1266), considering all patients. Nonetheless, a significant difference between CTVs delineated by CT and PET/CT was found when only head and neck, lung and lymphoma cases--representing more than 60% of the sample--were examined (112.5 and 80.7 ml, respectively; P=0.0053). We have shown that PET/CT use promotes significant changes in the CTV delineated for treatment of different tumors, modifying RT planning. Our data suggest that PET/CT has a good potential for optimizing RT treatment planning, especially in head and neck, lung, and lymphoma tumors.


Clinical, histopathological and immunohistological study of lymphoid disorders in the parotid gland of patients with Sjögren's syndrome.
Sjögren's syndrome is a chronic autoimmune systemic disease characterized by polyglandular tissue destruction, leading to keratoconjunctivitis sicca and xerostomia. These patients have 44-fold increased risk of developing salivary gland lymphoma, of which 80% are marginal zone (MALT) type. Having in mind that criteria for distinguishing benign lymphoepithelial lesions from MALT lymphoma are obscure, the aim of this study was to provide practical information that could be integrated into diagnostic practice. Among 32 parotidectomies, 27 cases were identified as having benign lymphoepithelial disorders and 5 cases low grade MALT lymphoma. Histological sections were stained routinely with hematoxylin and eosin (H&E and special stains. Immunohistochemical study was performed by LSAB2 method, by using primary antibodies for CD20, CD3, Kappa and Lambda light chains and Cytokeratin (Dako Denmark). The 27 patients with Sjögren's sialoadenitis (22 women and 5 men), and 5 patients with MALT lymphoma (only women) were included in this analysis. According to the Ann Harbor Classification, all patients with MALT lymphoma had stage IE. Both groups of patients had an indolent clinical course, except permanent, rapid parotid enlargement in the patients with MALT lymphoma. Histologically, the periductal lymphoid infiltrate, gradually extended to the acini, completely replacing them by a sea of polyclonal lymphocytes, immunoblasts, germinal centers and plasma cells (confirmed immunohistochemically), but sparing the ducts and preserving lobular appearance. The histological feature of salivary gland MALT lymphoma included heterogeneous B-cell infiltrate that totally or subtotally had effaced the normal glandular structure. Malign lymphoepithelial lesions, representing infiltration of the ductal and epithelial structures by monoclonal neoplastic B-cells, positive for CD20, were highlighted by antibody to cytokeratin. The optimal diagnosis of salivary gland MALT lymphoma requires careful integration of clinical, morphological and immunohistochemical results.


Synthesis and biological evaluation of heat-shock protein 90 inhibitors: geldanamycin derivatives with broad antiviral activities.
Previous studies have suggested that geldanamycin (GA) inhibits the replication of several viruses in vitro. Here, we aimed to synthesize and evaluate the antiviral activity of 17-amino-17-demethoxygeldanamycin derivatives. A series of 17-substituted and 17-,19-disubstituted GA derivatives were screened for antiviral activities against eight different viral strains, including herpesvirus, hepatitis virus, retrovirus and picornavirus. Most of the tested compounds showed inhibitory activity against the viruses and showed reduced cytotoxicity in vitro as compared with the parent compound GA. In vivo efficacy evaluation results showed that compound 6 noticeably inhibited duckling hepatitis B virus DNA replication in duckling serum after oral administration. Viral rebound did not occur after termination of the treatment. The modified GA derivatives also showed median lethal dose values that were higher than that of the parent GA in mice intraperitoneally treated with the study compounds. Targeting heat-shock protein 90 could be a new antiviral approach that is not prone to the development of drug resistance. The 17-amino-17-demethoxygeldanamycin derivatives could be novel agents with potential antiviral activity.


HES 130/0.4 impairs haemostasis and stimulates pro-inflammatory blood platelet function.
Hydroxyethyl starch (HES) solutions are widely used for volume replacement therapy but are also known to compromise coagulation, impair renal function and increase long-term mortality. To test the hypotheses that HES 130/0.4 has fewer adverse effects than HES 200/0.5 and exerts anti-inflammatory properties, we compared the effects of HES 130/0.4, HES 200/0.5 and saline on in vitro haemostasis and pro-inflammatory platelet function. Whole blood samples from healthy volunteers were mixed with 6% HES 130/0.4, 10% HES 200/0.5, or normal saline to achieve a final haemodilution rate of 10% or 40%. Haemostatic capacity was characterised by thromboelastography (ROTEM) and measurement for FXIIIa activity. Platelet activation and pro-inflammatory platelet functions were characterised by flow cytometry measuring the platelet activation marker CD62P and binding of fibrinogen to platelets as well as the formation of heterotypic platelet-leukocyte conjugates. Compared with saline, HES 130/0.4 dose-dependently impaired formation and firmness of the fibrin clot but did not affect the fibrin crosslinking activity of FXIIIa. At 40% but not at 10% haemodilution rate, HES 200/0.5 also increased platelet fibrinogen binding and both HES solutions increased expression of CD62P, the main receptor for platelet-leukocyte adhesion. HES 130/0.4 but not HES 200/0.5 increased formation of platelet-neutrophil conjugates and, to a lesser degree, platelet-monocyte conjugates. Our data demonstrate that HES 130/0.4 has similar adverse effects as HES 200/0.5. In particular, both types of HES impair coagulation capacity and stimulate, rather than attenuate, pro-inflammatory platelet function.


Isolation and distribution of West Nile virus in embryonated chicken eggs.
West Nile virus (WNV) was identified from domestic psittacine birds by inoculating embryonated chicken eggs. Most of the embryos died 5 days postinoculation; flavivirus was detected in some by negative-staining electron microscopy. Immunohistochemistry performed on the embryos and their supporting structures detected the WNV antigen mainly in the chorioallantoic membrane, regardless of the inoculation route or passage number. WNV antigen was also found in the embryonic muscle (both skeletal and smooth muscles) and in multifocal areas of the skin. WNV was not detected in the viscera of the embryo or yolk sac. This study provides evidence of isolation and identification of WNV via embryonated chicken eggs.


Direct induction of DNA hypermethylation in sea urchin embryos by microinjection of 5-methyl dCTP stimulates early histone gene expression and leads to developmental arrest.
The role of DNA methylation in gene inactivation has been studied extensively in vertebrates but it is not clear whether it serves similar functions in other organisms. We devised a novel approach to induce hypermethylation of both endogenous and injected DNA in the sea urchin Lytechinus pictus in order to study the effect of DNA methylation on gene expression in this invertebrate. By injecting 5-methyl dCTP either alone or together with a cloned DNA construct into fertilized sea urchin eggs, replicating DNA became hypermethylated from the random incorporation of the methylated nucleotide in place of cytosine during DNA synthesis. During subsequent rounds of replication, the injected 5-methyl dCTP became depleted but methylation at CpG sites was still elevated presumably due to the action of a methyltransferase enzyme. Using this approach, we studied the effect of hypermethylation on two members of the sea urchin multigene family, the early H2B and the late H2B genes. De novo methylation was shown to occur at known cis-regulatory regions of the genes. The effect of methylation on gene activity was probed using RNase protection assay. Methylation resulted in increased early H2B histone gene expression but had no effect on late H2B histone gene expression. These results demonstrate that methylation does not necessarily inactivate genes in the sea urchins as previously thought. Interestingly, the development of embryos injected with 5-methyl dCTP typically was arrested at the blastula stage, and analysis of the genomic DNA extracted from injected embryos showed a significant increase in the endogenous methylation content. These data suggest that perturbation of methylation patterns in developmental sea urchin embryos may be responsible for the developmental arrest through altering the gene expression pattern.


Genetic and molecular properties of human and rat renin-binding proteins with reference to the function of the leucine zipper motif.
The presence of a leucine zipper motif was recognized in the deduced amino acid sequences of human and rat renin-binding proteins (RnBPs) on cloning and sequence analysis of the RnBP cDNAs. The in vitro synthesized RnBPs, with the respective cDNAs, formed heterodimers with porcine renin and homodimers. On comparison of these properties with those of porcine RnBP, the leucine zipper motif was suggested to be a functional domain common to animal RnBPs. In addition to the motif, a hydrophobic domain adjacent to the motif and 10 cysteine residues were also well conserved in the three RnBPs. Moreover, about 85% of their amino acid sequences were identical. The RnBP mRNAs were expressed in the kidneys as the same size of 1.5-kb and the genes are suggested to exist as single copies in the genomes. Despite the high similarities in genetic and molecular properties, the molecular weights of human and rat RnBPs were 43,000, which is 1,000 larger than that of porcine RnBP. The immunoreactivities of human and rat RnBPs toward anti-porcine RnBP antiserum were 88 and 8% that of porcine RnBP, respectively, and the affinities of the two RnBPs for porcine renin were remarkably less than that of porcine RnBP. Moreover, the human and rat RnBP homodimers were partly dissociated under the conditions under which porcine RnBP existed as a dimer. These results indicate distinct differences in the molecular properties among the three RnBPs, in spite of their being highly similar structurally and functionally.


Lessons on sustainability for community health projects.
In the Gambia a community-based strategy was tested, in which a traditional snack food was promoted as a dietary supplement to improve women's nutrition during pregnancy. The results suggest how community nutrition programmes can be designed so as to favour sustainability. By and large, the lessons learned are also applicable to other types of community health programme.


Phosphorylation of insulin receptor substrate-1 on multiple serine residues, 612, 632, 662, and 731, modulates insulin action.
Okadaic acid has been described previously as being a negative regulator of insulin signaling, as it inhibits insulin stimulation of glucose transport. In addition, this drug induces on insulin receptor substrate-1 (IRS-1) a decrease in tyrosine phosphorylation, concomitantly with an increase in serine/threonine phosphorylation. The present work was aimed at the identification of the serine/threonine residues that, upon phosphorylation, might be involved in modulating insulin signaling. To this end, we studied double-point mutants of IRS-1, in which serines 612/632 and 662/731 were replaced with alanine. These are four plausible sites of phosphorylation by mitogen-activated protein kinases and are in the immediate proximity of tyrosine residues, which are potential sites of interaction with phosphatidylinositol 3-kinase Src homology 2 domains. Using transient expression in 293 EBNA cells, we demonstrate that serines 612, 632, 662, and 731 and mitogen-activated protein kinases are not involved in the okadaic acid effect on IRS-1. Rather, these serines appear to play a role in modulating basal and insulin-stimulated IRS-1 tyrosine phosphorylation, association of IRS-1, with p85, and phosphatidylinositol 3-kinase activity in the IRS-1.p85 immune complex, since mutation of these sites enhances these events. Our findings suggest the existence of an IRS-1 desensitization mechanism resulting from serine/threonine phosphorylation, occurring at least on serines 612, 632, 662, and 731.


Oocyte cryopreservation in the setting of a vascular endothelial growth factor (VEGF)-producing paraneoplastic syndrome: a case report and review of literature.
Many reproductive aged women with new oncologic diagnoses choose to undergo emergency oocyte or embryo cryopreservation prior to initiating potentially gonadal toxic oncologic therapies. Ovarian hyperstimulation syndrome (OHSS) is a potential complication of these treatments and can be particularly dangerous in these patients due to their underlying medical illness and by delaying lifesaving oncology treatment. This case report details a multipronged approach to OHSS prevention in a patient with a paraneoplastic syndrome defined by overproduction of vascular endothelial growth factor (VEGF), which is also primarily responsible for OHSS. A 29 year old nulligravid woman was diagnosed with a rare multisystem paraneoplastic syndrome (Polyradiculoneuropathy, organomegaly, endocrinopathy, monoclonal plasma cell disorder and skin changes, known as POEMS) and presented for fertility preservation consultation prior to autologous stem cell transplant. She successfully underwent oocyte cryopreservation without complications due to aggressive OHSS prophylaxis with both a dopamine agonist and aromatase inhibitor during and after stimulation and a gonadotropin releasing hormone agonist (GnRH-A) for final oocyte maturation, without delay in her subsequent oncology treatment. This is the first report of a patient with POEMS undergoing controlled ovarian hyperstimulation (COH). Oocyte cryopreservation was successful and without complications due to a combination of prophylactic measures against OHSS (cabergoline, letrozole and GnRH-A trigger) and close collaboration between reproductive endocrinology and oncology. This case demonstrates the use of combined measures in targeting VEGF overproduction and OHSS risk during COH.


Corollary discharge inhibition of wind-sensitive cercal giant interneurons in the singing field cricket.
Crickets carry wind-sensitive mechanoreceptors on their cerci, which, in response to the airflow produced by approaching predators, triggers escape reactions via ascending giant interneurons (GIs). Males also activate their cercal system by air currents generated due to the wing movements underlying sound production. Singing males still respond to external wind stimulation, but are not startled by the self-generated airflow. To investigate how the nervous system discriminates sensory responses to self-generated and external airflow, we intracellularly recorded wind-sensitive afferents and ventral GIs of the cercal escape pathway in fictively singing crickets, a situation lacking any self-stimulation. GI spiking was reduced whenever cercal wind stimulation coincided with singing motor activity. The axonal terminals of cercal afferents showed no indication of presynaptic inhibition during singing. In two ventral GIs, however, a corollary discharge inhibition occurred strictly in phase with the singing motor pattern. Paired intracellular recordings revealed that this inhibition was not mediated by the activity of the previously identified corollary discharge interneuron (CDI) that rhythmically inhibits the auditory pathway during singing. Cercal wind stimulation, however, reduced the spike activity of this CDI by postsynaptic inhibition. Our study reveals how precisely timed corollary discharge inhibition of ventral GIs can prevent self-generated airflow from triggering inadvertent escape responses in singing crickets. The results indicate that the responsiveness of the auditory and wind-sensitive pathway is modulated by distinct CDIs in singing crickets and that the corollary discharge inhibition in the auditory pathway can be attenuated by cercal wind stimulation.


[Application of adsorption resin separation technique in traditional China medicine research].
Adsorption resin separation is one of the new technologies developed after ion exchange resin in the 60's of the 20th century. It was early used in the fields of the wastewater treatment, pharmaceutical industry, chemical engineering, analytical chemistry, clinical detection and treatment. In recent years, the technique of adsorption resin separation has been widely used in research on active constituent of traditional China medicine. The recent application was reviewed in this paper.


Effects of Cortical Cooling on Sound Processing in Auditory Cortex and Thalamus of Awake Marmosets.
The auditory thalamus is the central nexus of bottom-up connections from the inferior colliculus and top-down connections from auditory cortical areas. While considerable efforts have been made to investigate feedforward processing of sounds in the auditory thalamus (medial geniculate body, MGB) of non-human primates, little is known about the role of corticofugal feedback in the MGB of awake non-human primates. Therefore, we developed a small, repositionable cooling probe to manipulate corticofugal feedback and studied neural responses in both auditory cortex and thalamus to sounds under conditions of normal and reduced cortical temperature. Cooling-induced increases in the width of extracellularly recorded spikes in auditory cortex were observed over the distance of several hundred micrometers away from the cooling probe. Cortical neurons displayed reduction in both spontaneous and stimulus driven firing rates with decreased cortical temperatures. In thalamus, cortical cooling led to increased spontaneous firing and either increased or decreased stimulus driven activity. Furthermore, response tuning to modulation frequencies of temporally modulated sounds and spatial tuning to sound source location could be altered (increased or decreased) by cortical cooling. Specifically, best modulation frequencies of individual MGB neurons could shift either toward higher or lower frequencies based on the vector strength or the firing rate. The tuning of MGB neurons for spatial location could both sharpen or widen. Elevation preference could shift toward higher or lower elevations and azimuth tuning could move toward ipsilateral or contralateral locations. Such bidirectional changes were observed in many parameters which suggests that the auditory thalamus acts as a filter that could be adjusted according to behaviorally driven signals from auditory cortex. Future work will have to delineate the circuit elements responsible for the observed effects.


Complications of acute rhinosinusitis in The Netherlands.
Despite the evidence demonstrating that antibiotics are of little benefit in acute rhinosinusitis (ARS), GPs continue to prescribe them, possibly in an attempt to prevent potentially dangerous complications. In this study, we present epidemiological data about the incidence, course and severity of such complications in the Netherlands. This retrospective cohort study included all patients hospitalized in The Netherlands in 2004 with a complication of ARS. Records were made of the symptoms of ARS and the complication, demographics, medical history, medical treatment preceding hospitalization, diagnostic techniques, therapeutic management, course and outcome. Forty-seven patients with 48 complications (16 intracranial and 32 orbital) were included. In the intracranial group (mean age 35.9 years), six patients had been treated with oral antibiotics prior to hospitalization. While hospitalized, all patients were treated with intravenous antibiotics and 15 underwent surgery. Eight patients recovered fully after treatment, three patients had residual symptoms and three patients died (missing data: 2). Of the 31 patients with orbital complications (mean age 17.4 years), 14 received oral antibiotics before admission. While hospitalized, all patients were treated with intravenous antibiotics and 13 underwent surgery. Twenty-seven patients recovered fully and two had residual symptoms (missing data: 2). Severe ARS complications occur in an otherwise healthy population in an estimated 1:12,000 paediatric and 1:32,000 adult cases in the Netherlands. Our study suggests that antibiotic treatment of ARS in general practice does not play a role in preventing complications.


Posterior ischemic optic neuropathy associated with migraine.
Posterior ischemic optic neuropathy (PION) is an uncommon form of optic nerve ischemia that results from damage to the intraorbital, intracanalicular, or intracranial optic nerve. It has been reported perioperatively, in association with systemic vasculitis, and in the nonsurgical setting with no identifiable cause. Review of the literature reveals only 2 patients with PION associated with migraine in a single report. We report a patient who developed PION in the setting of a migraine headache without any other identifiable risk factors.


Genomic profiling shows increased glucose metabolism in luminal B breast cancer.
We had previously reported a close association between pathological response and the maximum tumor standardized uptake value (SUVmax) measured by (18)F-fluorodeoxyglucose positron emission tomography prior to chemotherapy in estrogen receptor (ER)-positive breast cancer. We hypothesized that glucose hypermetabolism by luminal B tumors may result in chemotherapy responsiveness. Using a single-gene expression assay, TargetPrint® (Agendia) and a 70-gene expression classifier, MammaPrint® (Agendia), we divided 20 patients with ER-positive primary breast cancer into luminal A and luminal B subtypes and compared the tumor SUVmax value between the two groups. A significantly higher SUVmax was measured for luminal B tumors (n=10; mean±SD, 7.6±5.6) than for luminal A tumors (n=10; mean±SD, 2.6±1.2; p=0.01). Glucose hypermetabolism could help predict intrinsic subtyping and chemotherapy responsiveness as a supplement to ER, progesterone receptor, HER2, and Ki-67 histochemical scores.


Permeability of intact and dechorionated eggs of the Anopheles mosquito to water vapor and liquid water: a comparison with Drosophila.
As an initial step in the development of cryopreservation methods for Anopheles gambiae embryos, we determined the permeability of intact and dechorionated A. gambiae embryos to water and ethylene glycol and compared those permeabilities with those of Drosophila melanogaster embryos. Most studies were conducted on embryos allowed to develop for 8 h at 26 degrees C or 15 h at 17 degrees C. Intact A. gambiae embryos are some 50 times more permeable to water vapor than are D. melanogaster embryos; e.g., when air dried, half of 15 h/17 degrees C A. gambiae embryos become severely dehydrated in 3. 5-4.0 min, whereas comparably aged D. melanogaster embryos require approximately 4 h or more. Comparable differences between the two species exist with respect to the loss of liquid water under an osmotic driving force. After 90 min in 0.75 M sucrose in 0.26 osmolal D-20 Drosophila medium, 25-40% of intact A. gambiae embryos underwent extensive shrinkage, whereas D. melanogaster embryos showed no shrinkage. The chorion of both species can be removed by a 2.5-min exposure to 50% household bleach (Clorox). Dechorionation increases the rate of water loss two- to sixfold during air drying in both species and during exposure to hyperosmotic sucrose in A. gambiae. Dechorionated D. melanogaster embryos, however, show no shrinkage in hyperosmotic sucrose even after 6 h. The water permeability of dechorionated A. gambiae embryos thus remains many-fold higher than that of dechorionated D. melanogaster embryos. Although moderately permeable to water, neither intact nor dechorionated A. gambiae embryos are permeable to ethylene glycol, as evidenced by the failure of dehydrated embryos to reexpand in 120-180 min in that solution.


A stop-smoking telephone help line that nobody called.
The purpose of this study was to evaluate the reach of a smoker's hotline that provided a variety of services to over 2100 health maintenance organization members. Formative evaluation procedures identified topics of concern, and repeated promotions advertised the service via multiple channels. Excluding a special giveaway promotion, an average of less than three calls per month were made during the 33 months of operation. To be cost-effective, smoker's hotlines should be offered to large populations and should be consistently and intensively publicized.


Efficacy of soluble IL-4 receptor for the treatment of adults with asthma.
IL-4 mediates important proinflammatory functions in asthma, including induction of the IgE isotype switch, increased expression of vascular cell adhesion molecule 1 and promotion of eosinophil transmigration across the endothelium, stimulation of mucus production, and T(H)2 lymphocyte differentiation, leading to release of IL-4, IL-5, IL-9, and IL-13. The current study evaluated the therapeutic potential of inhaled recombinant human soluble interleukin-4 receptor (IL-4R) as an IL-4 antagonist. This study was a randomized, double-blind, placebo-controlled study in 62 subjects involving 12 once weekly nebulizations of 0.75, 1.5, or 3.0 mg of IL-4R or placebo. During screening, subjects documented dependence on inhaled corticosteroids by an exacerbation in asthma induced by one or two 50% dose reductions at 2-week intervals. After restabilization for 2 weeks on the dose above which their asthma flared, inhaled steroids were discontinued, patients were randomized, and study medication was started on day 0. IL-4R was well tolerated. Efficacy was demonstrated by a decline in FEV(1) observed in the placebo group (-0.4 L and -13% predicted), which did not occur in the group receiving 3.0 mg of IL-4R (-0.1 L and -2% predicted; P =.05 over the 3-month treatment period). Daily patient-measured morning FEV(1) also demonstrated a significant decline in the placebo group (-0.5 L and -18% predicted), which did not occur in the group receiving 3.0 mg of IL-4R (-0.1 L and -4% predicted; P =.02 over the 3-month treatment period). The efficacy of IL-4R was further confirmed by the absence of increase in asthma symptom scores in the group receiving 3.0 mg of IL-4R (Delta 0.1) compared with that seen in the placebo group (Delta 1.4 over 1 month; P =.07). Study discontinuation for asthma exacerbation was not significantly different between groups (placebo, 56%; 3.0 mg of IL-4R, 47%; P = not significant). These promising data suggest that IL-4R is safe and effective in the treatment of moderate persistent asthma.


Legislative priorities 2008 National Association for Home Care and Hospice.
Home care and hospice providers seek a number of legislative reforms in order to more effectively address the challenges they face. Following are the top-ranked legislative priorities for 2008, along with specific recommendations on the issues, as chosen on by the membership of the National Association for Home Care & Hospice, and voted by its board of directors at its February 2008 meeting.


Rapid access cardiology--a nine year review.
Rapid access cardiology clinics provide a rapid assessment and evaluation of patients with suspected chest pain, heart failure or cardiac arrhythmia. Longitudinal data regarding the attendances and clinical diagnoses at rapid access clinics is lacking and may provide insight into their efficiency and role in providing a comprehensive referral service for primary care. Data regarding total number of attendances and diagnoses were retrospectively analysed from databases collated by nurse specialists and physicians undertaking rapid access clinics at Charing Cross Hospital between April 2002 and March 2011. A total of 12 000 patients were seen over the 9-year time period. Demand for Rapid Access Chest Pain Clinic and Rapid Access Arrhythmia Clinic has shown a steady rise since 2007 whereas attendances at Rapid Access Heart Failure Clinic have remained constant. The proportion of patients diagnosed with significant cardiac disease across all rapid access clinics has not changed significantly. The most commonly diagnosed arrhythmia was atrial fibrillation (15% of all attendances). Referrals have increased to our service which suggests that demand for a rapid access cardiology service remains high and the proportion of patients diagnosed with significant cardiac disease has remained constant. Despite considerable motivation towards providing primary based care this has not occurred in our local area and we believe this shows that our model, based in secondary care to be both efficient and accessible to local GPs and patients.


FGF-2 deficiency does not influence FGF ligand and receptor expression during development of the nigrostriatal system.
Secreted proteins of the fibroblast growth factor (FGF) family play important roles during development of various organ systems. A detailed knowledge of their temporal and spatial expression profiles, especially of closely related FGF family members, are essential to further identification of specific functions in distinct tissues. In the central nervous system dopaminergic neurons of the substantia nigra and their axonal projections into the striatum progressively degenerate in Parkinson's disease. In contrast, FGF-2 deficient mice display increased numbers of dopaminergic neurons. In this study, we determined the expression profiles of all 22 FGF-ligands and 10 FGF-receptor isoforms, in order to clarify, if FGF-2 deficiency leads to compensatory up-regulation of other FGFs in the nigrostriatal system. Three tissues, ventral mesencephalon (VM), striatum (STR) and as reference tissue spinal cord (SC) of wild-type and FGF-2 deficient mice at four developmental stages E14.5, P0, P28, and adult were comparatively analyzed by quantitative RT-PCR. As no differences between the genotypes were observed, a compensatory up-regulation can be excluded. Moreover, this analysis revealed that the majority of FGF-ligands (18/22) and FGF-receptors (9/10) are expressed during normal development of the nigrostriatal system and identified dynamic changes for some family members. By comparing relative expression level changes to SC reference tissue, general alterations in all 3 tissues, such as increased expression of FGF-1, -2, -22, FgfR-2c, -3c and decreased expression of FGF-13 during postnatal development were identified. Further, specific changes affecting only one tissue, such as increased FGF-16 (STR) or decreased FGF-17 (VM) expression, or two tissues, such as decreased expression of FGF-8 (VM, STR) and FGF-15 (SC, VM) were found. Moreover, 3 developmentally down-regulated FGFs (FGF-8b, FGF-15, FGF-17a) were functionally characterized by plasmid-based over-expression in dissociated E11.5 VM cell cultures, however, such a continuous exposure had no influence on the yield of dopaminergic neurons in vitro.


Spinal cord MRI using multi-array coils and fast spin echo. I. Technical aspects and findings in healthy adults.
It is time-consuming to detect intrinsic spinal cord lesions in multiple sclerosis (MS) by MRI using conventional surface coils and T2-weighted spin-echo pulse sequences. Multi-array coils and fast spin-echo pulse sequences permit the generation of high-resolution T2-weighted sagittal images of the whole spinal cord in about 5 minutes. Using these advances, we found an area of high signal within the cord in only 1/45 (2%) healthy subjects aged 18 to 72 years, whereas 26% of those who underwent brain imaging had cerebral white matter abnormalities. Degenerative vertebral column changes, especially in the cervical region, were present in 64% and were associated with cord compression in 11%. Cord cross-sectional areas in mm2, measured from axial gradient-echo images, were usually highly reproducible and showed a significant correlation with the subject's height. We conclude that (1) MRI signal abnormalities within the spinal cord may be more specific for MS than cerebral white matter lesions, especially in subjects over 50 years old; (2) asymptomatic degenerative changes in the vertebral column are common, even in younger adults; (3) measurement of cord cross-sectional area should allow accurate quantitation of the degree of atrophy in MS and other spinal cord diseases; and (4) multi-array coils and fast spin echo represent an important advance in MRI of the spinal cord.


[Beta-blockers and statins in the context of asthma].
Asthma is a disease with a complex pathogenesis and differentiated clinical picture with airway inflammation in its background. Many cells and cell-released substances are engaged in the course of the disease. The basic treatment strategy in asthma is based on chronic administration of inhaled glucocorticosteroids (with a strong anti-inflammatory effect) and beta2-adrenoreceptor agonists (bronchodilatory effect). Much attention has been recently paid to the effects of other medicines on mechanisms important in the pathogenesis of asthma, including beta-blockers and statins. Many researchers have suggested a potentially useful role of some beta-blockers in chronic asthma therapy, particularly considering their effect on the pharmacodynamics of beta receptors in the bronchi. Moreover, statins, due to their anti-inflammatory and immunomodulatory effects, can also be useful in the management of asthma.


[Progress of study on the demodex blepharitis].
Demodex blepharitis is an infectious ocular disease which is common, but always to be neglected. There were a few related reports about this disease in China. The pathogenic mechanism of demodex blepharitis is not clear at present. Literatures held different ideas about its diagnosis, therapeutic methods and curative effect. This review will summarize the new development of demodex blepharitis in order to provide references for its basic and clinical researches.(Chin J Ophthalmol, 2016, 52: 315-320).


The crystal structure of the signal recognition particle Alu RNA binding heterodimer, SRP9/14.
The mammalian signal recognition particle (SRP) is an 11S cytoplasmic ribonucleoprotein that plays an essential role in protein sorting. SRP recognizes the signal sequence of the nascent polypeptide chain emerging from the ribosome, and targets the ribosome-nascent chain-SRP complex to the rough endoplasmic reticulum. The SRP consists of six polypeptides (SRP9, SRP14, SRP19, SRP54, SRP68 and SRP72) and a single 300 nucleotide RNA molecule. SRP9 and SRP14 proteins form a heterodimer that binds to the Alu domain of SRP RNA which is responsible for translation arrest. We report the first crystal structure of a mammalian SRP protein, that of the mouse SRP9/14 heterodimer, determined at 2.5 A resolution. SRP9 and SRP14 are found to be structurally homologous, containing the same alpha-beta-beta-beta-alpha fold. This we designate the Alu binding module (Alu bm), an additional member of the family of small alpha/beta RNA binding domains. The heterodimer has pseudo 2-fold symmetry and is saddle like, comprising a strongly curved six-stranded amphipathic beta-sheet with the four helices packed on the convex side and the exposed concave surface being lined with positively charged residues.


Morphological changes among hippocampal dentate granule cells exposed to early kindling-epileptogenesis.
Temporal lobe epilepsy is associated with changes in the morphology of hippocampal dentate granule cells. These changes are evident in numerous models that are associated with substantial neuron loss and spontaneous recurrent seizures. By contrast, previous studies have shown that in the kindling model, it is possible to administer a limited number of stimulations sufficient to produce a lifelong enhanced sensitivity to stimulus evoked seizures without associated spontaneous seizures and minimal neuronal loss. Here we examined whether stimulation of the amygdala sufficient to evoke five convulsive seizures (class IV or greater on Racine's scale) produce morphological changes similar to those observed in models of epilepsy associated with substantial cell loss. The morphology of GFP-expressing granule cells from Thy-1 GFP mice was examined either 1 day or 1 month after the last evoked seizure. Interestingly, significant reductions in dendritic spine density were evident 1 day after the last seizure, the magnitude of which had diminished by 1 month. Further, there was an increase in the thickness of the granule cell layer 1 day after the last evoked seizure, which was absent a month later. We also observed an increase in the area of the proximal axon, which again returned to control levels a month later. No differences in the number of basal dendrites were detected at either time point. These findings demonstrate that the early stages of kindling epileptogenesis produce transient changes in the granule cell body layer thickness, molecular layer spine density, and axon proximal area, but do not produce striking rearrangements of granule cell structure.


The contribution of nonlocal electro-opto-thermal interaction to single molecule nonlinear Raman enhancement.
we develop a precise modelling where nonlocal electro-opto-thermal interactions are comprehensively included for the analysis of nonlinear Raman enhancement and plasmonic heating. An interaction enhancement factor G(IEF) is introduced to quantify the coupling between the electromagnetic field and the temperature field which is rarely considered in the estimation of Raman enhancement. For the case of isolated single nanosphere, G(IEF) can be up to ten, indicating a thermal origin which well explains the observed temperature rise, shortened blinking period, and the nonlinearly enhanced Raman cross-section. For the case of nanodimer, the suppression of plasmon heating was analyzed, demonstrating the great capability to mitigate biomolecular degradation and blinking.


Spontaneous singleton and twin pregnancy in two patients with polycystic ovary syndrome and type 2 diabetes following treatment with metformin combined with rosiglitazone.
Two women, a Saudi Arab and a Caucasian, aged 28 and 41 years, had a long history of polycystic ovary syndrome (PCOS) and primary infertility, which was refractory to various treatment modalities. Both later developed type 2 diabetes, managed initially with metformin; rosiglitazone was added later to improve glycaemic control. Both conceived spontaneously, resulting in singleton and twin pregnancies giving birth to healthy babies. The combination of rosiglitazone with metformin may result in improved fertility with ovulation and spontaneous conception in women with PCOS and type 2 diabetes. Randomized controlled trials are needed to assess the feasibility of this combination to improve fertility in women with PCOS. However, in the absence of any safety data, those women, following counselling, should be warned against the possibility of spontaneous pregnancy and contraceptive measures should be discussed and offered where appropriate.


Ambient air arsenic levels along the Texas-Mexico border.
The Texas-Mexico border region is one of the most rapidly developing areas in the state. Unprecedented economic and demographic growth is placing stress on a system already overburdened with air, water, waste, and public health problems. This paper reports the results of an initial survey of the past and present ambient air levels of arsenic along the Texas side of the border. Ambient air arsenic levels have increased over time, exhibit seasonal variations, and are higher in specific areas when compared to the rest of the state. Five of the 20 areas in Texas having the highest 24-hour levels of arsenic are found along the border. In areas where emissions have been significantly reduced at the source, arsenic levels remained high in the environment for over two years. This provides a potential continuum for human exposure. The results of this study emphasize the need for bi-national development and implementation of air quality control programs along the Texas-Mexico border.


Paracoccidioides brasiliensis: attenuation of yeast cells by gamma irradiation.
Paracoccidioides brasiliensis is the agent of paracoccidioidomycosis, the most prevalent mycosis in Latin America, and currently there is no effective vaccine. The aim of this study was to attenuate the yeast form of P. brasiliensis by gamma irradiation for further studies on vaccine research. Paracoccidioides brasiliensis (strain Pb 18) cultures were irradiated at doses between 0.5 and 8.0 kGy. After each dose the viability, reproductive ability and protein metabolism were evaluated. The comparison between the antigenic profile of irradiated and control yeast was made by Western blot and the virulence evaluated by the inoculation in C(57)Bl/J6 mice. At 6.5 kGy the yeast lost its reproductive capacity. The viability and the incorporation of [L-(35)S]-methionine were the same in control and up to 6.5 kGy irradiated cells, but 6.5 kGy-irradiated yeast secreted 40% less proteins. The Western blot profile was clearly similar in control and 6.5 kGy-irradiated yeast. No colony-forming unit (CFU) could be recovered from the tissues of the mice infected with the radioattenuated yeast. We concluded that for P. brasiliensis yeast it is possible to find a dose in which the pathogen loses its reproductive ability and virulence, while retaining its viability, metabolic activity and the antigenic profile.


In vitro activity of echinocandins against 562 clinical yeast isolates from a Romanian multicentre study.
The study presents the echinocandin susceptibility profile of a multi-centre collection of pathogenic yeast isolates from Romanian tertiary hospitals. The 562 isolates were identified using ID32C strips, MALDI-TOF MS and DNA sequencing. Minimal inhibitory concentrations (MICs) of caspofungin (CAS), micafungin (MCA), and anidulafungin (ANI) were assessed and interpreted according to EUCAST guidelines. Minimal fungicidal concentrations (MFC) were determined by plating content from the clear MIC wells. The activity was considered fungicidal at MFC/MIC ≤ 4. The three echinocandins had strongly correlated MICs and high percentages of MIC essential agreement. Most often, MCA had the lowest MICs, followed by CAS and ANI. Against C. parapsilosis and C. kefyr, CAS had the lowest MIC values. The MIC50 values were between 0.03 and 0.25 mg/l, except C. parapsilosis. The MIC90 values were usually one dilution higher. MFCs and MICs were weakly correlated. ANI and MCA had the lowest MFC values. The MFC50 values were between 0.06 and 0.5 mg/l, except C. parapsilosis, C. guilliermondii, and C. dubliniensis. The MFC90 values were usually two dilutions higher. Based on EUCAST breakpoints, 47 isolates (8.4%) were resistant to at least one echinocandin, most often ANI. Most resistant isolates were of C. albicans, C. glabrata, and C. krusei. There were 17 isolates (3%) resistant to echinocandins and fluconazole and most belonged to the same three species. MCA and ANI had the highest rates of fungicidal activity. The high rates of echinocandin resistance and significant multidrug resistance make prophylaxis and empiric therapy difficult.


Pyruvate Decarboxylase from Zea mays L. : I. Purification and Partial Characterization from Mature Kernels and Anaerobically Treated Roots.
Pyruvate decarboxylase (PDC) was purified from mature, dry maize kernels and from roots of anaerobically treated maize seedlings and partially characterized. PDC was purified to a specific activity of 96 units per milligram protein from kernels and to 41 units per milligram protein from root. The subunit molecular masses were estimated to be 61,000 and 60,000 for kernel PDC and 59,000 and 58,000 for root PDC. The pH optimum for each enzyme was 5.8. Since the pH optimum is nearly one pH unit below the value reported for the cytoplasm of anaerobically metabolizing maize roots (pH 6.7 +/- 0.2), we investigated the effects of pH 5.8 and 6.6 on the cooperative kinetics observed for PDC from each source. The maximum Hill coefficients (n(H)) were much greater at each pH for the kernel PDC (pH 5.8, n(H) = 2.5 and pH 6.6, n(H) = 3.2) than for the root PDC (pH 5.8, n(H) = 1.4 and pH 6.6, n(H) = 1.8). The cooperative kinetics observed with respect to pyruvate were asymmetric. Potassium inhibited maize PDC and was competitive with pyruvate (root PDC K(i) = 16 millimolar and kernel PDC K(i) = 10 millimolar).


Interrelations between communicative behaviors at the outset of speech: parents as observers.
The Hebrew Parent Questionnaire for Communication and Early Language (HPQ-CEL) was administered by 154 parents of Hebrew-speaking toddlers aged 1 ; 0 to 1 ; 3 (77 boys, 77 girls). The Questionnaire guided parents in observing and rating their toddlers in six contexts at home. The study aimed to identify inter-correlations between toddlers' non-linguistic behaviors that co-occur during the transition to speech. Seven communicative behaviors were extracted from the questionnaire data: Crying, Vocalizations, Collaboration with Adults, Pointing, Words, Joint Engagement in a Peek-a-Boo Game, and Triadic Interaction in Book Reading. Collaboration with Adults and Triadic Interaction in Book Reading yielded more significant correlations than other prelinguistic behaviors. Participation in social games and book-reading activities was associated with the toddlers' number of words at the period studied.


Gap junctional communication controls the overall endothelial calcium response to vasoactive agonists.
A cytosolic calcium (Ca(2+)(i)) increase is an important activation signal for the endothelium. We investigated whether interendothelial spreading of the Ca(2+) signal via gap junctions (GJs) plays a role for the overall Ca(2+)(i) increase in response to vasoactive agonists. In human umbilical vein endothelial cells (HUVECs), a Ca(2+)(i) increase (Fura2) in response to histamine or ATP occurred initially only in about 30% of the cells (initially responding cells) reflecting the cell fraction expressing H(1) or purinergic receptors (FACS/immunohistochemistry). In the remaining adjacent cells, Ca(2+)(i) increases occurred only after a delay of up to 5 s. Blockade of GJ communication (meclofenamic acid and heptanol, or H(2)O(2); verified by dye injection) did not affect responses in the initially responding cells but abolished the delayed Ca(2+)(i) response of the remaining adjacent cells. The resulting reduction in the global endothelial Ca(2+)(i) response significantly reduced the nitric oxide synthesis (assessed as cGMP levels). Similar Ca(2+)(i) results were obtained in the endothelium of freshly isolated mouse (C57BL/6) aortas stimulated with ATP. The receptor-independent Ca(2+)(i) response to ionomycin occurred simultaneously in all cells, regardless of GJ inhibition. In separate experiments, inhibition of the IP(3) receptor (xestospongin-C; 40, µmol/L) but not of the ryanodine receptor (ryanodine, 250 µmol/L) reduced the spread of the Ca(2+)(i) signal into adjacent cells over longer distances. The global Ca(2+)(i) response of the endothelium to agonists is determined decisively by the functionality of GJs, thus establishing a new role for GJs in controlling endothelial activity and vasomotor function.


Stabilization of neurotoxic Alzheimer amyloid-beta oligomers by protein engineering.
Soluble oligomeric aggregates of the amyloid-beta peptide (Abeta) have been implicated in the pathogenesis of Alzheimer's disease (AD). Although the conformation adopted by Abeta within these aggregates is not known, a beta-hairpin conformation is known to be accessible to monomeric Abeta. Here we show that this beta-hairpin is a building block of toxic Abeta oligomers by engineering a double-cysteine mutant (called Abetacc) in which the beta-hairpin is stabilized by an intramolecular disulfide bond. Abeta(40)cc and Abeta(42)cc both spontaneously form stable oligomeric species with distinct molecular weights and secondary-structure content, but both are unable to convert into amyloid fibrils. Biochemical and biophysical experiments and assays with conformation-specific antibodies used to detect Abeta aggregates in vivo indicate that the wild-type oligomer structure is preserved and stabilized in Abetacc oligomers. Stable oligomers are expected to become highly toxic and, accordingly, we find that beta-sheet-containing Abeta(42)cc oligomers or protofibrillar species formed by these oligomers are 50 times more potent inducers of neuronal apoptosis than amyloid fibrils or samples of monomeric wild-type Abeta(42), in which toxic aggregates are only transiently formed. The possibility of obtaining completely stable and physiologically relevant neurotoxic Abeta oligomer preparations will facilitate studies of their structure and role in the pathogenesis of AD. For example, here we show how kinetic partitioning into different aggregation pathways can explain why Abeta(42) is more toxic than the shorter Abeta(40), and why certain inherited mutations are linked to protofibril formation and early-onset AD.


[Expression of the membrane antigens in native and cryopreserved embryonic precursor cells].
Native and cryoconservated human embryonic liver cells at different stages of gestation (5, 6 and 7 weeks) have been studied. Tissue desagregation was performed by trypsinisation technique. A part of the samples was studied after their trypsinisation, another part - after 3-week storage. Cell viability of all samples was higher than 90%. CD34 antigen expression was highest in 5-week gestation samples. The tendency of the deacrease of CD34 antigen expression was seen in 7-week gestation samples. CD34 antigen expression was higher in the samples after their cryoconservation than in native ones. The CD34 antigen expression afer their cryoconservation was accompanied by an increase in numbers of cells expressing CD38, CD45RA, CD33, CD13 and CD14 antigens as well as markers of erythroid precursor cells -CD71, CD36 and CD45RO. Percentage of alpha-actin positive cells was higher in 6- and 7-week samples. An increase in alpha-actin positive cell fraction has been seen also in samples after their cryogenic conservation.


Dynamic holography using pixelated light modulators.
Dynamic holography using spatial light modulators is a very flexible technique that offers various new applications compared to static holography. We give an overview on the technical background of dynamic holography focusing on pixelated spatial light modulators and their technical restrictions, and we present a selection of the numerous applications of dynamic holography.


The stem cell pluripotency factor NANOG activates transcription with two unusually potent subdomains at its C terminus.
Embryonic stem cells are pluripotent progenitors for virtually all cell types in our body and thus possess unlimited therapeutic potentials for regenerative medicine. NANOG, an NK-2 type homeodomain gene, has been proposed to play a key role in maintaining stem cell pluripotency presumably by regulating the expression of genes critical to stem cell renewal and differentiation. Here, we provide the evidence that NANOG behaves as a transcription activator with two unusually strong activation domains embedded in its C terminus. First, we identified these two transactivators by employing the Gal4-DNA binding domain fusion and reporter system and named them WR and CD2. Whereas CD2 contains no obvious structural motif, the WR or Trp repeat contains 10 pentapeptide repeats starting with a Trp in each unit. Substitution of Trp with Ala in each repeat completely abolished its activity, whereas mutations at the conserved Ser, Gln, and Asn had relatively minor or no effect on WR activity. We then validated the activities of WR and CD2 in NANOG by constructing a reporter plasmid bearing five NANOG binding sites. Deletion of both WR and CD2 from NANOG completely eliminated its transactivation function. Paradoxically, whereas the removal of CD2 reduced NANOG activity by approximately 30-70%, the removal of WR not only did not diminish but actually enhanced its activity by approximately 50-100% depending on the cell lines analyzed. These data suggest that either WR or CD2 is sufficient for NANOG to function as a transactivator.


The choice of radiologic procedures in the diagnosis of breast disease.
We have reviewed the radiologic imaging procedures related to the investigation of breast lesions, particularly carcinoma. We discussed the choice of procedure in certain clinical situations and have made suggestions. Of the imaging methods, x-ray mammography is the most useful. Ultrasound is the next most useful; however, it is not acceptable for screening. The basis for breast cancer detection and diagnosis remains palpation and x-ray mammography. Ultrasound makes a substantial contribution to the diagnostic evaluation of certain lesions. Eventually, biopsy or surgery provides the final answer in those patients with possible lesions after these diagnostic procedures. In view of the high incidence of breast carcinoma, it would be useful for the practitioner dealing with these problems to establish for his practice a standard operating procedure for examination and referral of patients for breast screening and diagnosis as well as a system of follow-up to prevent patients from avoiding the screening and diagnostic process.


CHRONIC USE OF PROTON PUMP INHIBITORS AND THE QUANTITY OF G, D, AND ECL CELLS IN THE STOMACH.
Acid inhibition from chronic proton pump inhibitor use and a possible increase in gastrin can lead to changes in the regulation of hydrochloric acid production. However, it has not known whether such chronic use changes the presence of gastrin, delta, and enterochromaffin-like cells in the stomach or the relationship between gastrin and delta cells. To analyze the number of gastrin-producing gastrin cells, somatostatin-producing cells, and histamine-producing cells in patients who were chronic users of proton pump inhibitor, with or without related Helicobacter pylori infection. Biopsies from 105 patients, including 81 chronic proton pump inhibitor users (experimental group) and 24 controls, were processed immunohistochemically and subjected to counting of gastrin, delta, and enterochromaffin-like cells in high-magnification microscopic fields and in 10 glands. Gastrin cell, delta cell, and enterochromaffin-like cells counts were similar across the groups and appeared to be unaffected by Helicobacter pylori infection. The ratio between gastrin cells and delta cells was higher in the chronic users of proton pump inhibitor group than in controls. Chronic users of proton pump inhibitor does not affect gastrin cell, delta cell, and enterochromaffin-like cell counts significantly, but may alter the ratio between gastrin cells and delta cells.


[Multiomics study of HepG2 cell line proteome].
Current proteomic studies are generally focused on the most abundant proteoforms encoded by canonical nucleic sequences. Transcriptomic and proteomic data, accumulated in a variety of postgenome sources and coupled with state-of-art analytical technologies, allow to start the identification of aberrant (non-canonical) proteoforms. The main sources of aberrant proteoforms are alternative splicing, single nucleotide polymorphism, and post-translational modifications. The aim of this work was to estimate the heterogeneity of HepG2 proteome. We suggested multiomics approach, which combines transcriptomic (RNAseq) and proteomic (2DE-MS/MS) methods, as a promising strategy to explore the proteome.


Expressions of cytokine genes during development of autoimmune sialadenitis in MRL/lpr mice.
Local cytokine gene expression in vivo was analyzed by direct analysis of RNA obtained from salivary gland tissues of MRL/lpr mice with autoimmune sialadenitis. The expression of cytokine genes were assessed by the reverse-transcriptase polymerase chain reaction, and by immunohistochemical analysis. The expression of interleukin-1(IL-1)beta and tumor necrosis factor was detected before the onset of inflammatory lesions in the salivary glands of mice of 1 or 2 months of age, and IL-6 mRNA expression was clearly detected at the time of onset of typical autoimmune sialadenitis at 3 months of age in MRL/lpr mice, and was up-regulated with advancing age. These results suggest that the overexpression of these inflammatory cytokine genes is involved in the development and progression of organ-localized autoimmunity in the salivary glands of MRL/lpr mice.


Cyclosporine-A discourages ethanol intake in C57bl/6j mice: a preliminary study.
Remarkably high rates of abstinence occur among alcohol-dependent persons after a liver transplant. This was thought to be solely due to selection or other nonbiological factors. Observing sustained abstinence for many months to years after a liver transplant, however, we asked whether immunosuppressants might exert a biological effect that supported abstinence from ethanol. We hypothesized that cyclosporine-A-treated mice would drink less ethanol given a choice between water and 10% ethyl alcohol. After a 3-day ethanol conditioning period, C57bl/6j mice (n = 20) medicated with cyclosporine-A (50 mg/kg) and a control group of unmedicated mice (n = 20) injected with saline were given free access to either water or the ethanol solution. The medicated mice drank significantly less ethanol throughout the 11-day choice protocol. This was significant whether measured by mean alcohol consumption per day (p = .003) or by alcohol preference as a percent of total liquid intake per day (p = .03). The two groups did not differ significantly in total daily fluid consumption. These data suggest that cyclosporine-A lowers alcohol preference in mice. Mechanism of action, similar effects by other centrally acting immunosuppressants and translation to humans all remain to be studied. If fruitful, elucidating the actions of cyclosporine and other immunosuppressants that activate central nervous system receptors may illuminate the pathophysiology of alcohol use and addiction.


Trauma and alkali burns induce distinct patterns of cytokine gene expression in the rat cornea.
Cytokines such as the interleukins (IL) and tumor necrosis factor alpha (TNF alpha) have traditionally been associated with paracrine regulation of immune reactions. These proteins also have properties suggestive of functional roles in the inflammatory and reparative responses to tissue injury. In this study, mRNA levels for IL-1 alpha, IL-1 beta, IL-6, TNF alpha, interferon gamma, transforming growth factor beta 1, and CD4 were monitored in rat corneas at times from 1 hour through 2 weeks after incisional trauma or alkali burns. Transcripts for IL-1 alpha, TNF alpha, and TGF beta 1 were present in most corneal samples; whereas those for IFN gamma and CD4 were not detected. As early as 1 hour following either of these non-immunologic forms of injury, expression of IL-6 mRNA levels was induced. Only in corneas with alkali burns did IL-6 induction persist from days 1 through 7. The alkali-injured corneas also had markedly increased IL-1 beta mRNA levels from days 1 through 7. These observations indicate that cytokine mRNA is induced in the cornea by trauma without an apparent immunologic stimulus. Our data are consistent with the hypothesis that corneal tissues respond to different types of injury with different patterns of cytokine gene expression.


Effects of High-Intensity Interval Training and Moderate-Intensity Continuous Training on Cardiometabolic Risk Factors in Overweight and Obesity Children and Adolescents: A Meta-Analysis of Randomized Controlled Trials.
Background: The purpose of this review was to compare the effectiveness of high-intensity interval training (HIIT) and moderate-intensity continuous training (MICT) on cardiometabolic risk factors of obese children and adolescents. Methods: Relevant studies published in PubMed, MEDLINE and Web of Science databases were searched. Only randomized controlled trials (RCTs) that examined the effect of HIIT and MICT on children and adolescents with obesity were included. Meta-analyses were conducted to determine the effect of HIIT on cardiometabolic risk factors using STATA software and potential moderators were explored (i.e., study duration, training modalities, work/rest ratio and work duration time). Results: Twelve RCTs involving 325 participants were included in the meta-analysis. HIIT showed more positive effects on maximal oxygen uptake (VO2max; SMD = 0.87, 95% CI: 0.39 to 1.35, p = 0.000) and systolic blood pressure (SBP; SMD = -0.64, 95% CI: -1.05 to -0.22, p = 0.003) than MICT. However, when compared with MICT, HIIT caused no significant differences in body weight, body mass index, body fat percentage, diastolic blood pressure and glycolipid metabolism markers. Furthermore, subgroup analysis showed that the effects of HIIT on VO2max and SBP were significantly different regarding protocol factors, such as modality, duration, training time, training settings, work/rest ratio and work duration. Conclusions: HIIT has a positive role in promoting cardiometabolic risk factors in obese children and adolescents. Moreover, when compared with MICT, HIIT had a more significant effect on improving cardiorespiratory fitness and systolic blood pressure. The factors of HIIT protocol had an important influence on the intervention effects of childhood obesity.