Insomnia Severity, Subjective Sleep Quality, and Risk for Obstructive Sleep Apnea in Veterans With Gulf War Illness.
Despite the fact that sleep disturbances are common in veterans with Gulf War Illness (GWI), there has been a paucity of published sleep studies in this veteran population to date. Therefore, the present study examined subjective sleep quality (assessed with the Pittsburgh Sleep Quality Index), insomnia severity (assessed with the Insomnia Severity Index), and risk for obstructive sleep apnea (assessed with the STOP questionnaire) in 98 Gulf War veterans. Veterans with GWI, defined either by the Kansas or Centers for Disease Control and Prevention criteria, had greater risk for obstructive sleep apnea (i.e., higher STOP scores) than veterans without GWI. This difference persisted even after accounting for potentially confounding demographic (e.g., age, gender) and clinical variables. Veterans with GWI, defined by either the Kansas or Centers for Disease Control and Prevention criteria, also had significantly greater insomnia severity and poorer sleep quality than veterans without GWI (p < 0.05), even after accounting for potentially confounding variables. Furthermore, there were significant, positive correlations between insomnia severity, subjective sleep quality, and GWI symptom severity (p ≤ 0.01). In stepwise linear regression models, insomnia severity significantly predicted GWI status over and above demographic and clinical variables. Together these findings provide good rationale for treating sleep disturbances in the management of GWI.


Two distinct pituitary cell lines from mouse intermediate lobe tumors: a cell that produces prolactin-regulating factor and a melanotroph [seecomments].
The intermediate lobe (IL) of the pituitary produces a PRL-regulating factor (PRF). Targeted tumorigenesis, using the POMC promoter ligated to SV40 large T antigen (Tag), generated transgenic mice that develop IL tumors with PRF activity. Our goal was to establish and characterize a PRF-producing cell line. Two cell lines, which differ markedly in size and morphology, were independently developed from IL tumors and designated mIL5 and mIL39. These cells are transformed, as judged by rapid proliferation, low serum requirements, and generation of secondary tumors in nude mice. RT-PCR revealed that mIL39, but not mIL5 cells, express POMC and dopamine D2 receptors, typical of a melanotroph phenotype. Although mIL5 cells originated from an IL tumor, they do not express messenger RNA for SV40 Tag. The bioassay for PRF used GH3 cells stably transfected with the PRL promoter ligated to a luciferase reporter gene (GH3/luc). Coculture of mIL5 with GH3/luc cells induced cell-density dependent increases in PRL gene expression and release, whereas mIL39 cells showed negligible PRF activity. Incubation of GH3/luc cells with conditioned media from mIL5, but not mIL39 cells, stimulated PRL gene expression and release up to 10-fold. Coculture of mIL5 cells with primary rat anterior pituitary cells stimulated PRL, but not GH, release. Fractionation of mIL5 cell extracts by reverse phase HPLC resolved PRF activity into one major and one minor peak. In conclusion, we have developed two novel and distinct cell lines from mouse intermediate lobe tumors. The first reported melanotroph cell line, mIL39, could provide a valuable model for studying dopaminergic regulation of POMC gene expression and release. In contrast, the mIL5 cells do not express POMC, D2 receptors, or SV40 Tag and appear to have been immortalized by a spontaneous mutation(s). These cells produce and secrete a potent PRF and could be used for the purification and biochemical characterization of PRF.


Corneal morphogenesis during development and diseases.
To review the use of genetically modified mouse lines for elucidating corneal morphogenesis during embryonic development and diseases. Transgenesis and gene-targeting techniques were used to create doxycycline-inducible mouse models (tet-On) to express transgenes or ablation of LoxP-modified genes or both in corneal cells, e.g., epithelial cells, and keratocytes and periocular mesenchymal cells of neural crest origin. Two driver mouse lines, i.e., Krt12-rtTA and Kera-rtTA, were created, which express reverse tetracycline transcription activator (rtTA) in corneal epithelial cells and keratocytes, respectively. Bitransgenic (Krt12-rtTA/tet-o-FGF7) and triple transgenic mice (Krt12rtTA/tet-o-Cre/Ctnnb1 and Kera-rtTA/tet-o-Cre/Ctnnb1) were obtained through cross-breeding tet-o-FGF7, tet-o-Cre, and Ctnnb1 mice. On doxycycline induction, overexpression of FGF7 by corneal epithelial cells of bitransgenic Krt12-rtTA/tet-o-FGF7 mice caused nuclear translocation of beta-catenin and epithelium hyperplasia resembling human ocular surface squamous neoplasia; in triple transgenic mice (Krt12rtTA/tet-o-Cre/Ctnnb1), constitutive nuclear translocation of mutant beta-catenin (loss of exon 3) leads to hyper proliferation of corneal epithelial cells; in comparison of expression of beta-catenin mutant protein by migrating, periocular mesenchymal cells of Kera-rtTA/tet-o-Cre/Ctnnb1 caused eyelid malformation. Use of genetically modified mice is of great value to study the pathophysiology of ocular surface defects resulting from genetic mutations.


Pulmonary hyalinizing granuloma detected in a family member after confirmation of tuberculosis in his father.
Pulmonary hyalinizing granuloma (PHG) is an uncommon lung disease that usually presents as bilateral multiple nodules, and more rarely as a solitary nodule. An exaggerated immune response to antigenic stimuli resulting from infection or an autoimmune process has been suggested as the cause of PHG. Here, we describe a rare case of solitary PHG that was detected in a family member after tuberculosis had been confirmed in his father, without any background of infectious disease or autoimmune abnormality.


Shark Ig light chain junctions are as diverse as in heavy chains.
We have characterized a small family of four genes encoding one of the three nurse shark Ig L chain isotypes, called NS5. All NS5 cDNA sequences are encoded by three loci, of which two are organized as conventional clusters, each consisting of a V and J gene segment that can recombine and one C region exon; the third contains a germline-joined VJ in-frame and the fourth locus is a pseudogene. This is the second nurse shark L chain type where both germline-joined and split V-J organizations have been found. Since there are only two rearranging Ig loci, it was possible for the first time to examine junctional diversity in defined fish Ig genes, comparing productive vs nonproductive rearrangements. N region addition was found to be considerably more extensive in length and in frequency than any other vertebrate L chain so far reported and rivals that in H chain. We put forth the speculation that the unprecedented efficiency of N region addition (87-93% of NS5 sequences) may be a result not only of simultaneous H and L chain rearrangement in the shark but also of processing events that afford greater accessibility of the V or J gene coding ends to terminal deoxynucleotidyltransferase.


Product Development of Natural Fibre-Composites for Various Applications: Design for Sustainability.
New product development review article aims to consolidate the principles and current literature on design for sustainability to seek the field's future direction. In this point of view, the design for sustainability methods can be established under the idea of sustainability in dimensions of ecology, economy and social pillars. Design for sustainability concept is implemented in concurrent engineering, including concept, embodiment and detail design processes. Integrating sustainability in engineering designs is crucial to producing greener products, system innovation, and services aligned with current market demand. Currently, many concurrent engineering studies related to natural fibre-reinforced polymer composites associated with sustainability enhance the application of design for sustainability techniques by professional designers. However, the current literature is scarce in bridging the design for sustainability concept with concurrent engineering during the design development stage, and these areas should be further developed. Several other future research directions, such as the need for aligning with principles and applications, along with exploring the relationships between the design for sustainability techniques and views of sustainability, are presented in this review paper.


An AGAMOUS MADS-box protein, SlMBP3, regulates the speed of placenta liquefaction and controls seed formation in tomato.
AGAMOUS (AG) MADS-box transcription factors have been shown to play crucial roles in floral organ and fruit development in angiosperms. Here, we isolated a tomato (Solanum lycopersicum) AG MADS-box gene SlMBP3 and found that it is preferentially expressed in flowers and during early fruit developmental stages in the wild-type (WT), and in the Nr (never ripe) and rin (ripening inhibitor) mutants. Its transcripts are notably accumulated in the pistils; transcripts abundance decrease during seed and placental development, increasing again during flower development. SlMBP3-RNAi tomato plants displayed fleshy placenta without locular gel and extremely malformed seeds with no seed coat, while SlMBP3-overexpressing plants exhibited advanced liquefaction of the placenta and larger seeds. Enzymatic activities related to cell wall modification, and the contents of cell wall components and pigments were dramatically altered in the placentas of SlMBP3-RNAi compared with the WT. Alterations in these physiological features were also observed in the placentas of SlMBP3-overexpressing plants. The lignin content of mature seeds in SlMBP3-RNAi lines was markedly lower than that in the WT. RNA-seq and qRT-PCR analyses revealed that genes involved in seed development and the biosynthesis of enzymes related to cell wall modification, namely gibberellin, indole-3-acetic acid, and abscisic acid were down-regulated in the SlMBP3-RNAi lines. Taking together, our results demonstrate that SlMBP3 is involved in the regulation of placenta and seed development in tomato.


Age-related changes and secular trends in hand bone size.
The aim of the present study was to evaluate age- and sex-related changes in the size and shape of long hand bones in a large Chuvashian cohort using cross-sectional and longitudinal study designs. The data were gathered in 1994 (557 individuals) and 2002 (513 individuals). The latter sample included 260 individuals that were studied only during the second expedition, and 253 individuals who were previously investigated in 1994. Statistical analyses included a maximum likelihood-based model-fitting technique and a t-test comparison. We found evidence for secular trend of hand bone size in both males and females within the Chuvashian population. In males, the length and total area of the long hand bones were greater in younger individuals, but mid-shaft bone width remained almost the same in individuals born at different periods of the last century. In females, the length of the hand bones and total bone area remained unchanged in women born after 1937. However, bone mid-shaft width gradually decreased in women born after 1940. Therefore, we argue that, at least within the Chuvashian population, there is a secular trend towards a more gracile appendicular skeleton in both males and females.


Curricular and Co-curricular Coverage of Leadership Competencies and the Influence of Extracurricular Engagement on Leadership Development.
Objective. To evaluate coverage of leadership-related competencies in a Doctor of Pharmacy (PharmD) curriculum and the impact of co-curricular and extracurricular experiences on students' leadership perceptions and self-efficacy. Methods. Course syllabi were used to comprehensively map the PharmD curriculum to 11 competencies related to the Leadership CAPE outcome. A survey was developed and administered to all first year through fourth year pharmacy (P1-P4) students to evaluate their leadership experience and engagement, and to assess their attitudes and self-efficacy in 11 leadership competencies. Descriptive statistics were used to evaluate student level of engagement in leadership activities and the Mann-Whitney U test was used to compare students' attitudes and self-efficacy based on the extent of leadership engagement. Results. Curriculum mapping revealed that all competencies were covered in at least one course at the introductory or reinforce levels. There were 362 students (68% of all PharmD students) who completed the survey. When responses from students who reported active engagement in pharmacy student organizations (defined as e-board member or chair of committee, N=142 or 39% of respondents) were compared to the rest of the cohort, statistically significantly better attitudes and self-efficacy were seen in 7 of 11 competencies. Perceptions and self-efficacy of the 72 students (19.9%) who reported active engagement in non-pharmacy organizations did not significantly differ from those not actively involved in any organizations. Conclusion. The leadership competencies curriculum mapping exercise was useful in identifying gaps in coverage and depth of coverage, inconsistencies in course objectives accurately reflecting coverage of leadership topics and competencies, and considerations that should be given to co-curricular leadership development. Active leadership engagement in pharmacy student organizations should be encouraged because of associated improved leadership perceptions and self-efficacy.


Organ and effective dose evaluation in coronary angiography by using a 320 MDCT based on in-phantom dose measurements with TLDs.
The purpose of this study is to estimate the organ and effective dose (E) from computed tomography coronary angiography (CTCA) on a 320-MDCT scanner. Radiation dose was estimated for the prospectively ECG-gated CTCA in a male phantom. A total of 451 chips of thermoluminescent dosimeter were implanted in the phantom for measuring the organ doses. The effective doses were calculated using measured organ doses and the tissue-weighting factors. The dose length product (DLP) values were recorded and used to develop the conversion coefficient k = 0.017 mSv•(mGy•cm)(-1) (E/DLP). In a 3-beat acquisition, the organ doses ranged from 0.24 to 71.55 mGy, and the doses in breast, bone surface, oesophagus, and lung were higher than 20 mGy. The effective doses in 2-beat and 3-beat acquisition were estimated to be 14.3 and 24.3 mSv. More beats of acquisition led to higher radiation dose. The reported k values for chest CT scan can be used to roughly estimate the E value from CTCA for 320 MDCT.


Primary open-angle glaucoma is associated with sleep apnea syndrome.
The etiology of primary open-angle glaucoma remains unclear. Various risk factors, including vascular abnormalities, have been associated with this disease. Sleep-associated diseases, like sleep apnea syndrome, might also represent a risk factor. Sleep apnea syndrome is characterized by repetitive upper airway obstructions during sleep, inducing hypoxia and sleep disruption with the risk of cardiovascular and neurological sequelae. In this study, we determined the prevalence of sleep apnea syndrome in primary open-angle glaucoma patients. Overnight transcutaneous finger oximetry was performed in 30 consecutive patients having primary open-angle glaucoma. We assessed the oximetry disturbance index during night sleep, a parameter used to diagnose sleep apnea syndrome and to grade its severity. Sleep apnea syndrome was more prevalent among primary open-angle glaucoma patients compared to normal historic controls of the same age and sex distribution (chi(2) = 9.35, d.f. = 3, p < 0.025). The oximetry disturbance index grade was significantly larger in the primary open-angle glaucoma group compared to normal controls (U = 3, 352, p = 0.01). According to the oximetry disturbance index, 20% (6/30) of primary open-angle glaucoma patients had sleep apnea syndrome. Primary open-angle glaucoma is associated with sleep apnea syndrome. Early recognition and treatment of sleep apnea syndrome are important to avoid cardiovascular and neurological complications.


Osteitis fibrosa cystica generalizata with adult T-cell leukaemia: a case report.
We report on a 62-year-old female initially suffering from extreme pain in both her lower extremities. Plain radiographs revealed multiple osteolytic lesions. Laboratory analyses indicated high levels of serum calcium and parathyroid hormone related protein (PTHrP) and detected HTLV-1 antibody. Histological examination of biopsied specimens which were obtained from osteolytic lesions indicated osteitis fibrosa cystica (OFC) without tumour cells. After the initial biopsy, a subcutaneous mass developed in the left thigh. Microsections of the biopsied mass disclosed adult T-cell leukaemia (ATL) neoplastic cells. Immunohistochemical findings revealed that PTHrP secreted from ATL neoplastic cells induced the OFC.


Epidemiology of human listeriosis.
During the 1980s, investigation of several large epidemics of listeriosis confirmed that transmission of L. monocytogenes in food causes human disease. Progress in laboratory detection and subtyping of the organism has enhanced our ability to compare human and environmental isolates of L. monocytogenes. Transmission by foodborne organisms is now recognized as causing both epidemic and sporadic listeriosis. Continued study of dietary risk factors associated with listeriosis is needed in order to develop dietary recommendations for the expanding population at increased risk of disease. Current research application of new molecular methods to the study of L. monocytogenes may improve the ability to diagnose pregnancy-associated disease and permit the rapid detection and control of L. monocytogenes in the food supply.


[Adverse reactions to contrast media for intravenous use. A comparison between ionic and nonionic media].
The purpose of this study was to show that the nonionic contrast media of low osmolarity are better tolerated than ionic media of high osmolarity, which to date are most frequently used in the authors hospital. We found that adverse reaction occurred in 80% of the group in which nonionic contrast medium was used. The authors conclude that nonionic, low osmolarity contrast media significantly reduce the frequency of adverse reactions and increase their safe use.


Atrial natriuretic factor R1 receptor from bovine adrenal zona glomerulosa: purification, characterization, and modulation by amiloride.
The atrial natriuretic factor (ANF) R1 receptor from bovine adrenal zona glomerulosa was solubilized with Triton X-100 and purified 13,000-fold, to apparent homogeneity, by sequential affinity chromatography on ANF-agarose and steric exclusion high-performance liquid chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining of the purified receptor preparation in the absence or presence of dithiothreitol revealed a single protein band of Mr 130,000. Affinity cross-linking of 125I-ANF to the purified receptor resulted in the labeling of the Mr 130,000 band. The purified receptor bound ANF with a specific activity of 6.8 nmol/mg of protein, corresponding to a stoichiometry of 0.9 mol of ANF bound/mol of Mr 130,000 polypeptide. Starting with 500 g of adrenal zona glomerulosa tissue, we obtained more than 500 pmol of purified receptor with an overall yield of 9%. The purified receptor showed a typical ANF-R1 pharmacological specificity similar to that of the membrane-bound receptor. The homogeneous Mr 130,000 receptor protein displayed high guanylate cyclase activity [1.4 mumol of cyclic GMP formed min-1 (mg of protein)-1] which was not stimulated by ANF. This finding supports the notion that the ANF binding and the guanylate cyclase activities are intrinsic components of the same polypeptide. Finally, the purified ANF-R1 receptor retained its sensitivity to modulation by amiloride, suggesting the presence of an allosteric binding site for amiloride on the receptor protein.


[A case report of chemoradiotherapy combined with S-1 responding to squamous cell carcinoma of the anal canal].
A 79-year-old woman was reffered to our hospital with the chief complaint of hematochezia. Type-2 tumor was found on anal canal by colonoscopy, and pathologic examination revealed a poorly differentiated squamous cell carcinoma. Computed tomography of the abdomen demonstrated obturatory node metastasis. The patient was diagnosed as having squamous cell carcinoma of the anal cana (l cStage III). After four months from chemoradiation (66 Gy/33 Fr plus S-1), the ulcer side was improved completely to epithelization, and abdominal CT scan showed a remarkable reduction of obturatory node metastasis. She was obtained a complete response. Now the patient is disease-free for ten months after chemoradiation.


Construction of GSH-triggered cationic fluoropolymers as two-in-one nanoplatforms for combined chemo-gene therapy.
Combined chemo-gene therapy has become a promising approach for enhanced anti-cancer treatment efficacy. However, effective co-delivery of therapeutic genes and drugs into target cells and tissues remains a major challenge. In this work, a GSH-responsive cationic fluoropolymer PSSF was designed as a co-delivery platform and synthesized by introducing a perfluorinated chain into a low-molecular weight PEI-based cationic polymer through a disulfide bond. PSSF exhibits good ability for drug and gene loading, as well as fast drug release in a GSH-rich environment. Gene transfection assay revealed that PSSF could deliver both model genes and the p53 gene into tumor cells smoothly with good protein expression, while maintaining good biocompatibility. It was also demonstrated that PSSF could simultaneously deliver the p53 gene and DOX into the HeLa cells efficiently and realize fast release of DOX. In vitro and in vivo anti-tumor assays both demonstrated that the co-delivery system could inhibit tumor growth more effectively than individual gene or drug therapy. Histopathological analysis of major organs indicated negligible systemic toxicity of such synergistic therapy systems. This rationally designed co-delivery vector provides an effective platform for the development of gene-drug synergistic therapy.


Does any relationship exist between P-glycoprotein-mediated multidrug resistance and intracellular calcium homeostasis.
Multidrug resistance (MDR) of neoplastic tissue represents a real obstacle to the effective chemotherapy of cancer. Several mechanisms of MDR were identified, from which the over-expression and efflux activity of P-glycoprotein (P-gp) - a plasma membrane ATPase (ABCB1 member of ABC transporter family) - represents the most commonly observed reason for neoplastic disease chemotherapy malfunction. The process of P-gp-mediated MDR seems to be related to intracellular calcium homeostasis, at least indirectly, for the following reasons: i. substances blocking calcium influx through L-type of calcium channels like verapamil were often found to antagonize P-gp-mediated MDR; ii. calcium signal abnormalities were observed in cells over-expressing P-gp; iii. cells with P-gp-mediated MDR were often resistant to thapsigargin; iv. several differences in intracellular calcium localization were observed when P-gp-negative and P-gp-positive cells were compared; and v. differences in the contents of several proteins of the endoplasmic reticulum involved in calcium homeostasis were observed to be associated with P-gp over-expression. This current study represents an attempt to summarize the knowledge about the possible relationship between P-gp-mediated MRD and intracellular calcium homeostasis.


Irreversible inhibition of soybean lipoxygenase-1 by hydroperoxy acids as substrates.
Soybean lipoxygenase-1 was irreversibly inactivated by various peroxy acids containing a cis,cis-1,4-pentadiene group. Among these compounds, 15(S)-hydroperoxyeicosatetraenoic acid (15(S)-HPETE)2 was found to be the most effective in the inactivation of lipoxygenase. Although the prior exposure of 15(S)-HPETE to hemoglobin abolished the inhibitory effect of 15(S)-HPETE, the simultaneous inclusion of hemoglobin potentiated the inactivation of lipoxygenase by 15(S)-HPETE alone. Interestingly, the potentiating effect of hemoglobin was observed only in the incubations with peroxy acids possessing the cis,cis-1,4-pentadiene. In either the presence or the absence of hemoglobin, it was commonly observed that the enzyme inactivation, which was maximal at pH 10, was significantly protected by tocopherol, but neither by mannitol nor ethanol, and that the inclusion of arachidonic acid or linoleic acid prevented the enzyme inactivation. Based on these results, it is suggested that the selective inactivation of lipoxygenase by these peroxy acids may be due to unstable intermediates produced from hydroperoxy acids bound to the active site of lipoxygenase.


Optimal Illumination Patterns for Fluorescence Tomography.
Fluorescence tomography has become increasingly popular for detecting molecular targets for imaging gene expression and other cellular processes in vivo in small animal studies. In this imaging modality, multiple sets of data are acquired by illuminating the animal surface with different excitation patterns, each of which produces a distinct spatial pattern of fluorescence. This work addresses one of the most intriguing, yet unsolved, problems of fluorescence tomography, which is to determine how to optimally illuminate the animal surface so as to maximize the information content in the acquired data. The key idea of this work is to parameterize the illumination pattern and to maximize the information content in the data by improving the conditioning of the Fisher information matrix. We formulate our problem as a constrained optimization problem. We compare the performance of different geometric illumination schemes with those generated by this optimization approach using the Digimouse atlas.


TLR5-mediated phosphoinositide 3-kinase activation negatively regulates flagellin-induced proinflammatory gene expression.
Epithelial cells detect motile pathogens via TLR5 ligation of flagellin, resulting in rapid induction of antibacterial/proinflammatory gene expression. Although such flagellin-induced gene expression is quite transient, likely to avoid the negative consequences of inflammation, little is known regarding the molecular mechanisms that mediate its shutdown. We hypothesized that, analogous to the case for TLR4, phosphoinositide 3-kinase (PI3K) might negatively regulate TLR5 signaling. However, because PI3K is an essential positive mediator of some pathways of TLR-mediated gene expression, the opposite hypothesis was also considered. Herein, we observed that flagellin stimulation of epithelial cells indeed induced rapid (<30 min) PI3K activation, as evidenced by Akt phosphorylation, via a TLR5-mediated mechanism. Blockade of PI3K with wortmannin resulted in marked enhancement of flagellin-induced gene expression as assessed by measuring levels of inducible NO synthase, IL-6, and IL-8. Such enhancement of gene expression by PI3K inhibition correlated with prolonged activation of MAPK (p38 and ERK1/2) and was ablated under MAPK inhibition. Such effect of inhibiting PI3K with wortmannin was mimicked by the PI3K inhibitor LY294002, and, conversely, a constitutively active PI3K prevented p38 activation in response to flagellin. Last, to test the significance of these results in vivo, we measured flagellin-induced gene expression in PI3K knockout mice. PI3K-null mice displayed increased levels of flagellin-induced serum IL-6, KC (IL-8 homolog), and nitrite as compared with heterozygous littermates. Thus, TLR5's rapid activation of PI3K serves to limit MAPK signaling, thus limiting proinflammatory gene expression and reducing the potential negative consequences of proinflammatory gene expression.


[Thrombolysis in acute occlusion of cerebral blood vessels].
Thrombolysis may achieve recanalization in cases of occlusion of the cerebral vessels. If therapy is initiated in good time, development of cerebral infarction may be at least partially prevented. Thrombolytic treatment was performed in 14 patients at the Wagner-Jauregg Hospital within a period of one year. Urokinase was given locally, while rtPA was applied locally and/or systemically. 4 patients had an occlusion of the internal carotid artery, 6 an occlusion of the middle cerebral artery, and 4 an occlusion of the basilar artery. Complete recanalization was achieved in 6 patients, partial recanalization in 4, and no recanalization in 4. The neurological outcome of the cases with complete recanalization was good with the exception of one patients who died. Partial recanalization resulted in a fair outcome in 2 patients, while the other 2 died. 3 out of the 4 patients in whom no recanalization was achieved died. Our findings show that this form of therapy may considerably improve the natural history of the disease, provided recanalization is achieved in good time. They encourage us to continue this form of therapy and to work at improving the therapeutic criteria.


Sonication is superior to scraping for retrieval of bacteria in biofilm on titanium and steel surfaces in vitro.
Low-virulence implant infections are characterized by bacterial colonization of the implant with subsequent biofilm formation. In these cases, soft tissue biopsies often prove to be culture negative. Consequently, detachment of the causative adherent bacteria is crucial for correct microbiological diagnosis. Using an in vitro model, we compared 4 methods of biofilm sampling from metal surfaces. Discs of titanium and steel were incubated in the presence of Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, and Propionibacterium acnes in Mueller Hinton broth. Non-adherent bacteria were removed by repeated rinsing of the discs. 10 parallels of each disc were subjected to 1 of 4 methods for bacterial recovery: (A) sonication of the discs, (B) scraping of the discs using surgical blades followed by streaking of the blades onto agar plates, (C) scraping of the discs followed by vortex mixing of the surgical blades, and (D) scraping of the discs followed by sonication of the surgical blades. Quantitative bacterial cultures were performed for each sampling method. With the exception of S. epidermidis on steel, sonication efficiently and reliably dislodged biofilm bacteria. The scraping methods employed did not detach bacteria embedded in biofilm. Scraping of metal surfaces is not an adequate method for sampling of biofilm bacteria in vitro.


Copper-catalyzed cyclization of iodo-tryptophans: a straightforward synthesis of pyrroloindoles.
Pyrroloindoles are a key structural motif found in a wide number of biologically active alkaloids. Intramolecular copper-catalyzed coupling of readily accessible 2-iodo-tryptophan derivatives occurs in excellent yield, affording a wide range of polysubstituted, enantioenriched tetrahydropyrrolo[2,3- b]indoles. Diketopiperazines are also suitable substrates for this cyclization reaction, which affords a straightforward entry to tetra- to hepta-polycyclic systems.


Lysozyme synthesis in osteoclasts.
Osteoclasts may or may not be directly related to monocytes and macrophages, but it is well established that these cell types share a number of features in common. In the present study we sought to extend this comparison by assessing lysozyme synthesis in osteoclasts, an enzyme known to be produced and secreted in large amounts by monocytes and macrophages. Our data show that freshly isolated chicken osteoclasts and osteoclasts in situ contain an abundant amount of lysozyme and correspondingly high steady-state levels of the enzyme's messenger RNA. Marrow macrophages, at various stages of in vitro maturation, also possess lysozyme mRNA but in amounts approximately two to four times lower than osteoclasts. These observations reaffirm the monocyte-macrophage nature of the osteoclast but raise questions about the function of the lysozyme in this cell. At present, the role of the lysozyme in osteoclast activity remains unexplained.


Exercise Intensity Modulates Glucose-Stimulated Insulin Secretion when Adjusted for Adipose, Liver and Skeletal Muscle Insulin Resistance.
Little is known about the effects of exercise intensity on compensatory changes in glucose-stimulated insulin secretion (GSIS) when adjusted for adipose, liver and skeletal muscle insulin resistance (IR). Fifteen participants (8F, Age: 49.9±3.6yr; BMI: 31.0±1.5kg/m2; VO2peak: 23.2±1.2mg/kg/min) with prediabetes (ADA criteria, 75g OGTT and/or HbA1c) underwent a time-course matched Control, and isocaloric (200kcal) exercise at moderate (MIE; at lactate threshold (LT)), and high-intensity (HIE; 75% of difference between LT and VO2peak). A 75g OGTT was conducted 1 hour post-exercise/Control, and plasma glucose, insulin, C-peptide and free fatty acids were determined for calculations of skeletal muscle (1/Oral Minimal Model; SMIR), hepatic (HOMAIR), and adipose (ADIPOSEIR) IR. Insulin secretion rates were determined by deconvolution modeling for GSIS, and disposition index (DI; GSIS/IR; DISMIR, DIHOMAIR, DIADIPOSEIR) calculations. Compared to Control, exercise lowered SMIR independent of intensity (P<0.05), with HIE raising HOMAIR and ADIPOSEIR compared with Control (P<0.05). GSIS was not reduced following exercise, but DIHOMAIR and DIADIPOSEIR were lowered more following HIE compared with Control (P<0.05). However, DISMIR increased in an intensity based manner relative to Control (P<0.05), which corresponded with lower post-prandial blood glucose levels. Taken together, pancreatic insulin secretion adjusts in an exercise intensity dependent manner to match the level of insulin resistance in skeletal muscle, liver and adipose tissue. Further work is warranted to understand the mechanism by which exercise influences the cross-talk between tissues that regulate blood glucose in people with prediabetes.


The lncRNA CCAT2 rs6983267 G allele is associated with decreased susceptibility to recurrent miscarriage.
Genetics might play various roles in susceptibility to recurrent miscarriage, and previous studies suggest that some gene polymorphisms might be associated with abortion and breast cancer onset. Colon cancer-associated transcript 2 (CCAT2) is a novel long noncoding RNA (lncRNA) transcript that might be correlated with susceptibility to multiple cancers, including breast cancer. However, whether lncRNA CCAT2 polymorphisms are related to susceptibility to recurrent miscarriage is unclear. We genotyped two lncRNA CCAT2 polymorphisms (rs6983267 and rs3843549) in 248 patients with recurrent miscarriage and 392 controls through a TaqMan real-time polymerase chain reaction assay, and the strength of each association was evaluated via 95% confidence intervals (CIs) and odds ratios (ORs). Our results showed that the rs6983267 G allele in lncRNA CCAT2 was associated with decreased susceptibility to recurrent miscarriage (TG vs. TT: adjusted OR = 0.603; 95% CI = 0.420-0.866; p = 0.0062; GG/TG vs. TT: adjusted OR = 0.620; 95% CI = 0.441-0.873; p = 0.0061). The combined analysis of the two protective polymorphisms (rs3843549 AA and rs6983267 TG/GG) revealed that individuals with two unfavorable alleles exhibited a lower risk of recurrent miscarriage than those with no or only one unfavorable allele (adjusted OR = 0.531; 95% CI = 0.382-0.739). Moreover, the decreased risk associated with the two protective alleles was most obvious in women aged less than 35 years (OR = 0.551; 95% CI = 0.378-0.8803; p = 0.0019) and in women with two to three miscarriages (adjusted OR = 0.466; 95% CI = 0.318-0.683; p < 0.0001). In conclusion, our study indicates that the rs6983267G allele might contribute to a decreased risk of recurrent miscarriage in the South Chinese population.


Visualization of sigma1 receptors in eyes by ex vivo autoradiography and in vivo positron emission tomography.
Sigma receptors are present on the neurons of the central nervous system and in peripheral organs. They have also been demonstrated in ocular tissues by in vitro membrane binding assays. We have investigated whether sigma(1) receptors can be demonstrated in rat eyes by ex vivo autoradiography using [(11)C]SA4503, a selective radioligand. We also tested whether in vivo positron emission tomography (PET) can be used to show sigma(1) receptors in rabbit eyes. In rats, a high accumulation of [(11)C]SA4503 was found in the iris-ciliary body and retina. A carrier-loading experiment showed that the receptor-specific binding of [(11)C]SA4503 was approximately 75% of the total binding in the brain. Sigma(1) receptors were also detected in the projecting terminals of the retina to the superior colliculus. PET showed radioactivity in the anterior segment including the iris-ciliary body and retina, and pretreatment or displacement by a sigma receptor ligand (haloperidol), suggested that the PET signal reflects radioligand-receptor binding. The high density of sigma(1) receptors in the iris-ciliary body and retina was confirmed by ex vivo autoradiography. In conclusion, the iris-ciliary body and retina are rich in sigma(1) receptors, and PET may be used to investigate the in vivo distribution of these neuroreceptors in the eye.


Getting the Sergeants on your side: the importance of interpersonal relationships and cultural interoperability for generating interagency collaboration between nurses and the police in custody suites.
In this article, I contribute to the literature around interagency collaboration, especially between law enforcement and health care, by reconciling the previous work of Sarah Charman (2014) with the interprofessional teamwork literature. Drawing upon a semi-structured interview-based study with 20 custody nurses working in English police custody suites (analysed using Framework Analysis), I explore the ways they are able to achieve interagency collaboration with a particular police officer, the Desk Sergeant. I argue that nurses accomplish interagency interoperability by interacting regularly with the Desk Sergeant, anticipating their needs and limiting their own goals to those that are commensurate with the Desk Sergeant's, notably providing information and avoiding deaths in custody. Such practices are similar to the strategies of 'Cultural Interoperability' noted by Charman (2014); however, this study also identifies that such strategies are only available once a successful working relationship has formed between Desk Sergeant and nurse, and as a result, similarly to the findings of interprofessional teamwork studies, the success or failure of attempts to collaborate across agencies is underpinned by interpersonal relations. The paper concludes by commenting on the importance of analysing both the interpersonal and organisation levels when studying collaboration.


Trends in US Emergency Department Visits for Mental Health, Overdose, and Violence Outcomes Before and During the COVID-19 Pandemic.
The coronavirus disease 2019 (COVID-19) pandemic, associated mitigation measures, and social and economic impacts may affect mental health, suicidal behavior, substance use, and violence. To examine changes in US emergency department (ED) visits for mental health conditions (MHCs), suicide attempts (SAs), overdose (OD), and violence outcomes during the COVID-19 pandemic. This cross-sectional study used data from the Centers for Disease Control and Prevention's National Syndromic Surveillance Program to examine national changes in ED visits for MHCs, SAs, ODs, and violence from December 30, 2018, to October 10, 2020 (before and during the COVID-19 pandemic). The National Syndromic Surveillance Program captures approximately 70% of US ED visits from more than 3500 EDs that cover 48 states and Washington, DC. Outcome measures were MHCs, SAs, all drug ODs, opioid ODs, intimate partner violence (IPV), and suspected child abuse and neglect (SCAN) ED visit counts and rates. Weekly ED visit counts and rates were computed overall and stratified by sex. From December 30, 2018, to October 10, 2020, a total of 187 508 065 total ED visits (53.6% female and 46.1% male) were captured; 6 018 318 included at least 1 study outcome (visits not mutually exclusive). Total ED visit volume decreased after COVID-19 mitigation measures were implemented in the US beginning on March 16, 2020. Weekly ED visit counts for all 6 outcomes decreased between March 8 and 28, 2020 (March 8: MHCs = 42 903, SAs = 5212, all ODs = 14 543, opioid ODs = 4752, IPV = 444, and SCAN = 1090; March 28: MHCs = 17 574, SAs = 4241, all ODs = 12 399, opioid ODs = 4306, IPV = 347, and SCAN = 487). Conversely, ED visit rates increased beginning the week of March 22 to 28, 2020. When the median ED visit counts between March 15 and October 10, 2020, were compared with the same period in 2019, the 2020 counts were significantly higher for SAs (n = 4940 vs 4656, P = .02), all ODs (n = 15 604 vs 13 371, P < .001), and opioid ODs (n = 5502 vs 4168, P < .001); counts were significantly lower for IPV ED visits (n = 442 vs 484, P < .001) and SCAN ED visits (n = 884 vs 1038, P < .001). Median rates during the same period were significantly higher in 2020 compared with 2019 for all outcomes except IPV. These findings suggest that ED care seeking shifts during a pandemic, underscoring the need to integrate mental health, substance use, and violence screening and prevention services into response activities during public health crises.


Fiber optic temperature sensor using sampled homodyne detection.
A method of homodyne detection using laser emission frequency modulation and sampling of the interferometer output signal is described for a temperature sensor using a Mach-Zehnder all-fiber interferometer configuration. Both ac and dc signal detection are possible with this scheme. Temperature changes over the 5-80 degrees C range are measurable with a resolution of 1.4 mK by fringe counting and electronically determining the slope of the phase change.


Automated array-based genomic profiling in chronic lymphocytic leukemia: development of a clinical tool and discovery of recurrent genomic alterations.
B cell chronic lymphocytic leukemia (B-CLL) is characterized by a highly variable clinical course. Recurrent chromosomal imbalances provide significant prognostic markers. Risk-adapted therapy based on genomic alterations has become an option that is currently being tested in clinical trials. To supply a robust tool for such large scale studies, we developed a comprehensive DNA microarray dedicated to the automated analysis of recurrent genomic imbalances in B-CLL by array-based comparative genomic hybridization (matrix-CGH). Validation of this chip in a series of 106 B-CLL cases revealed a high specificity and sensitivity that fulfils the criteria for application in clinical oncology. This chip is immediately applicable within clinical B-CLL treatment trials that evaluate whether B-CLL cases with distinct chromosomal abnormalities should be treated with chemotherapy of different intensities and/or stem cell transplantation. Through the control set of DNA fragments equally distributed over the genome, recurrent genomic imbalances were discovered: trisomy of chromosome 19 and gain of the MYCN oncogene correlating with an elevation of MYCN mRNA expression.


Unilateral cerebral polymicrogyria with ipsilateral cerebral hemiatrophy.
We evaluated six children in whom MR imaging showed unilateral cerebral polymicrogyria associated with ipsilateral cerebral atrophy and ipsilateral brain stem atrophy. The aim of this study was to clarify whether this disorder based on neuroimaging constitutes a new homogeneous clinical entity. The subjects were six children whose ages at the time of MR imaging ranged from 8 months to 11 years. Their clinical and MR features were analyzed. All of the children were born between 38 and 42 weeks gestation, without any significant perinatal events. Spastic hemiplegia and epilepsy were observed in all of the patients, and mental retardation was observed in four. The MR findings included unilateral cerebral polymicrogyria associated with ipsilateral cerebral hemiatrophy and ipsilateral brain stem atrophy in all patients. The ipsilateral sylvian fissure was hypoplastic in four patients. These patients showed relatively homogeneous clinical and neuroimaging features. Although the additional clinical features varied according to the site and the extent affected by the polymicrogyria, this disorder could constitute a new relatively homogeneous clinical entity.


Characteristics of indoor/outdoor particulate pollution in urban and rural residential environment of Pakistan.
Particulate pollution has emerged as a serious environmental health concern in Pakistan. The use of biomass fuels in traditional stoves produces high levels of indoor air pollutants. In Pakistan, 94% of rural and 58% of urban households depend on biomass fuel. This study investigates variations in indoor/outdoor concentrations of particulate matter during various activities for three different micro-environments in Pakistan. At a rural site, the average indoor/outdoor ratios for PM(10), PM(2.5), and PM(1), in kitchens using biomass fuels were 3.80, 4.36, and 4.11, respectively. A large variation was recorded in the mass concentration of particulate matter during cooking with concentrations in the range 4000-8555 microg/m(3). In a living room at a rural site, the average indoor/outdoor ratios for PM(10), PM(2.5), and PM(1) were 1.74, 2.49, and 3.01, respectively. At the urban site, the average indoor/outdoor ratios for the same size fractions were 1.71, 2.88, and 3.47, respectively. Cooking, cleaning and smoking were identified as principal contributors to the high indoor levels of particulate matter. This study showed considerably high concentrations of particulate matter, particularly in kitchens using biomass fuels, as compared to living areas. Thus women and children face the greatest exposure due to the amount of time they spend in the kitchen. In the developing world, particulate air pollution, both indoor and outdoor, is a substantial health hazard to the public. The very high concentrations of particulate matter in both rural and urban sites, particularly in kitchens using biomass fuels, emphasize the severity of this issue in Pakistan. Women and children are extensively at risk due to amount of time spent in kitchens. This state of affairs calls for a large-scale intervention to reduce the exposure to indoor air pollution.


Recovery 9 years post stroke with standardized electrical stimulation.
ABSTRACT This case report summarizes results of a new standardized evidence-based treatment protocol regarding neuromuscular electrical stimulation (NMES) and somatosensory stimulation (SS) for a chronic stroke patient. The protocol included treatment five times per week, up to 3 hr per day for 20 months. Treatment included 20 min active NMES followed by two to three hr of SS. Repetitive task practice activities were performed during SS for targeted muscle groups. Remarkable functional recovery was noted with all outcome measures, including the Chedoke-McMaster Stroke Assessment and the Action Research Arm Test, suggesting prolonged NMES and SS therapy should be investigated further.


Conceptual design of the tangentially viewing combined interferometer-polarimeter for ITER density measurements.
One of the systems planned for the measurement of electron density in ITER is a multi-channel tangentially viewing combined interferometer-polarimeter (TIP). This work discusses the current status of the design, including a preliminary optical table layout, calibration options, error sources, and performance projections based on a CO2/CO laser system. In the current design, two-color interferometry is carried out at 10.59 μm and 5.42 μm and a separate polarimetry measurement of the plasma induced Faraday effect, utilizing the rotating wave technique, is made at 10.59 μm. The inclusion of polarimetry provides an independent measure of the electron density and can also be used to correct the conventional two-color interferometer for fringe skips at all densities, up to and beyond the Greenwald limit. The system features five chords with independent first mirrors to reduce risks associated with deposition, erosion, etc., and a common first wall hole to minimize penetration sizes. Simulations of performance for a projected ITER baseline discharge show the diagnostic will function as well as, or better than, comparable existing systems for feedback density control. Calculations also show that finite temperature effects will be significant in ITER even for moderate temperature plasmas and can lead to a significant underestimate of electron density. A secondary role TIP will fulfill is that of a density fluctuation diagnostic; using a toroidal Alfvén eigenmode as an example, simulations show TIP will be extremely robust in this capacity and potentially able to resolve coherent mode fluctuations with perturbed densities as low as δn∕n ≈ 10(-5).


Redox properties of cytochrome c.
The redox properties of cytochromes (cyt) c, a ubiquitous class of heme-containing electron transport proteins, have been extensively investigated over the last two decades. The reduction potential (E degrees') is central to the chemistry of cyt c for two main reasons. First, E degrees' influences both the thermodynamic and kinetic aspects of the electron exchange reaction with redox partners. Second, this thermodynamic parameter is remarkably sensitive to changes in the properties of the heme and the protein matrix, and hence can be profitably used for the investigation of the solution chemistry of cyt c. This research area owes much to the exploitation of voltammetric techniques for the determination of E degrees' for metalloproteins, which dates back to the late 1970s. Since then, much effort has been devoted to the comprehension of the molecular factors that control E degrees' in cyt c, which include first coordination sphere effects on the heme iron, the interactions of the heme group with the surrounding polypeptide chain and the solvent, and also include medium effects related to the nature and ionic composition of the solvent, pH, the presence of potential protein ligands, and the temperature. This article provides an overview of the most significant advances made in this field recently.


Emphysematous Aortitis due to Clostridium septicum in an 89-Year-Old Female with Ileus.
Emphysematous aortitis is a rare but lethal form of infectious vasculitis. This condition was found incidentally on computed tomography of the chest during the evaluation of a patient presenting with pneumonia coincident with adynamic ileus. The patient did not have a history of malignancy. While colon cancer could not be ruled out, it is possible that ileus may have contributed to or resulted in bacterial translocation in this case. Appropriate investigations and empirical therapy against Clostridium septicum should be initiated in the presence of clinical and radiological findings suggestive of emphysematous aortitis.


Role of immunotherapy in stage III nonsmall cell lung cancer.
Despite aggressive treatment based on definitive chemoradiotherapy, 5-year overall survival in unresectable stage III nonsmall cell lung cancer remains poor (15-20%). The novel immunotherapy based on immune checkpoint inhibitors (ICIs) presents as the therapeutic 'Holly Grail' in lung cancer treatment. Preclinical models provide evidence of immunotherapy-radiotherapy (IM-RT) synergy. The exposure to ionizing radiation turns tumor in an in-situ vaccine, primes the innate immune system, increases immunotherapy efficacy by overcoming the immunosuppressive microenvironment of immune-resistant tumors and promotes a systemic, out-of-field antitumor T-cell-mediated response called abscopal effect. The immunomodulatory and abscopal effects of radiotherapy can be further enhanced by combining with systemic immunotherapies. The phase III START trial proved that liposomal vaccine - tecemotide (L-BLP25) administered as maintenance therapy after concurrent chemoradiotherapy (CRT) in LA-NSCLC prolongs survival. In the phase III PACIFIC trial consolidation with durvalumab, an anti-PDL-1 antibody, was associated with survival benefit in patients diagnosed with LA-NSCLC who responded to concurrent chemoradiotherapy. PACIFIC trial results are expected to definitely establish durvalumab as standard consolidation strategy in LA-NSCLC. Many clinical trials are ongoing in the field of immunoradiotherapy in LA-NSCLC to define the optimal conditions for this therapeutic combination.


Dual-mode interaction between quercetin and DNA-damaging drugs in cancer cells.
DNA-damaging drugs constitute standard chemotherapy regimen for advanced colorectal cancer. Here, the interactions between quercetin and 5-fluorouracil (5-FU), etoposide, and camptothecin were examined in cancer cells. HCT116 colorectal or PPC1 prostate cancer cells were treated with quercetin and the drugs. Clonogenicity assays, cell cycle profiles, and expressions of p53, p21, BAX, survivin and cyclin B1 proteins were used to examine the effects of the treatments. Quercetin synergistically inhibited the clonogenicity of the wild-type cells, but inhibited the cell cycle effects of all the drugs tested. In p53-null cells, the combination of low dose 5-FU with up to 6 μM quercetin promoted clonogenic survival. Treatment of p53-wild-type cells with 50 μM quercetin reduced drug-induced up-regulation of p53, p21 and BAX. The combination of quercetin and the drugs also reduced the levels of cyclin B1 and survivin proteins. While high doses of quercetin synergize with DNA-damaging agents, the effect of drug combination with quercetin is influenced by the effective doses and the p53 status of the cells.


Phosphorylation and dephosphorylation of Ser852 and Ser889 control the clustering, localization and function of PAR3.
Cell polarity is essential for various asymmetric cellular events, and the partitioning defective (PAR) protein PAR3 (encoded by PARD3 in mammals) plays a unique role as a cellular landmark to establish polarity. In epithelial cells, PAR3 localizes at the subapical border, such as the tight junction in vertebrates, and functions as an apical determinant. Although we know a great deal about the regulators of PAR3 localization, how PAR3 is concentrated and localized to a specific membrane domain remains an important question to be clarified. In this study, we demonstrate that ASPP2 (also known as TP53BP2), which controls PAR3 localization, links PAR3 and protein phosphatase 1 (PP1). The ASPP2-PP1 complex dephosphorylates a novel phosphorylation site, Ser852, of PAR3. Furthermore, Ser852- or Ser889-unphosphorylatable PAR3 mutants form protein clusters, and ectopically localize to the lateral membrane. Concomitance of clustering and ectopic localization suggests that PAR3 localization is a consequence of local clustering. We also demonstrate that unphosphorylatable forms of PAR3 exhibited a low molecular turnover and failed to coordinate rapid reconstruction of the tight junction, supporting that both the phosphorylated and dephosphorylated states are essential for the functional integrity of PAR3.


Persisting demand and supply gap for maternal and newborn care in eastern Uganda: a mixed-method cross-sectional study.
The slow progress in reducing maternal and newborn death in low and middle-income countries is attributed to both demand and supply-side factors. This study assessed the changes in maternal and newborn services in health facilities as well as demand for maternal and newborn health services in Eastern Uganda. The health assessment data were collected in August 2013 and September 2015 in the districts of Kamuli, Pallisa, and Kibuku. We purposively collected data on the availability of services from 40 health facilities that provided maternal and newborn services. In addition, we conducted 24 focus group discussions (FGDs) with women and men; and 18 key informant interviews (KIs) with health workers. On the supply side, most health facilities persistently lacked lifesaving medicines such as misoprostol, IV Ampicillin, IV Gentamycin, IV Metronidazole, Magnesium Sulphate, Ergometrine, Corticosteroids, ferrous Sulphate, Folic Acid, Combined ferrous, Benzyl penicillin, and Diazepam (IM or IV). Basic newborn equipment such as stethoscope, fetal scope, working baby scale, newborn suction devices, newborn resuscitation device, and thermometer were persistently not available in most of the health facilities. Binders for Kangaroo Mother Care, blanket to wrap newborn, baby warmer or heat lamp were persistently not available in at least 80% of the health facilities. Other equipment for the management of labor and abortions such as Manual vacuum aspirator for abortion care, blank partographs and vacuum extractor were not available in most of the health facilities including referral facilities at baseline and follow-up. On the demand side, the qualitative interviews exposed long distances and inadequate transport to the health facilities, inadequate information, poverty, and poor services at the health facilities as major factors that impede women to utilize/access maternal and newborn services. There are distinct influences on both demand and supply side, which restrain both health care uptake and its quality. The frequent disparity between the health facility readiness to provide services and the women readiness to utilize them needs to be addressed as the country intensifies its efforts to reduce maternal and newborn deaths through boosting facility deliveries.


Variations in prenatal screening in a US federal healthcare system: Same coverage, different options.
The Military Health System (MHS) is a federally funded organization that provides care to active duty service members and their beneficiaries. Our objective was to determine what methods of prenatal screening are used by military treatment facilities (MTFs), assess variations between institutions, and determine how practice patterns align with national recommendations. We surveyed all MTFs offering comprehensive prenatal care (n = 49). Departments were asked about aneuploidy screening options, availability of diagnostic testing, and carrier screening. In all, 43 MTFs (88%) completed the survey. Most (39/43) patients were stratified based on risk (predominantly maternal age at delivery and history). The most commonly offered test was combined 1st/2nd trimester screening (59%). Sixty percent routinely offered diagnostic testing, though less than half routinely offered microarrays. The majority offered universal carrier screening for cystic fibrosis (98%) and complete blood count with screening for thalassemias and hemoglobinopathies (88%). At the time of data collection, only five facilities (12%) had implemented spinal muscular atrophy carrier screening. Considerable heterogeneity exists in prenatal aneuploidy testing and carrier screening within the MHS. Standardized guidelines, protocols, and laboratory support would improve processes across the system. Additional resources including genetic counseling support and provider education are needed.


Refractive index sensor using microfiber-based Mach-Zehnder interferometer.
A simple and robust refractive index (RI) sensor based on a Mach-Zehnder interferometer has been demonstrated. A section of optical microfiber drawn from silica fiber is employed as the sensing arm. Because of the evanescent field, a slight change of the ambient RI will lead to the variation of the microfiber propagation constant, which will further change the optical length. In order to compensate the variation of the optical length difference, a tunable optical delay line (ODL) is inserted into the other arm. By measuring the delay of the ODL, the ambient RI can be simply demodulated. A high RI sensitivity of about 7159 μm/refractive index unit is achieved at microfiber diameter of 2.0 μm.


Cost effectiveness in the intensive care unit.
Effective policies to reduce true costs will require integrated information systems and demand behavioral changes from providers. A congenial environment must be created among medical educators, providers, vendors, and consumers if cost reduction is to be accomplished without compromising quality or access to critical care services. Physicians should do everything they believe may be of benefit for their patients, but we have an obligation to educate the public about the limitations of our art and the fact that "doing everything" is not always best for the patient or the grieving family. A significant method of controlling ICU costs is closely monitoring which patients are admitted and when they are discharged. Laboratory tests represent a source of cost reduction, and physicians must learn to order specific tests and not simply a battery of tests which includes the actual test desired. Limits should be placed on the tests that are ordered in terms of number and frequency. Improved efficiency of the utilization of resources should improve the care of our patients. The largest budget item of any or most critical care units is nursing; it is paramount that this essential and invaluable resource be utilized in a cost-effective manner. Diminishing unnecessary activity will both decrease complications and have salutary effects. Having more time to be with patients and their families will decrease our sense of failure and fulfill the important goal of caring. Physicians and nurses can return to thinking, assessing, and decision making instead of frenetically ordering, reacting, and intervening, which, we believe, accurately describes informational overload created by undue emphasis on high technology. In this way, we can respond to Fuch's exhortation that "physicians consider the possibility of contributing more by doing less." In responding, however, we must never forget that the societal, not merely the economic impact of medical care, is our principal consideration. We must first contribute more by achieving a greater understanding of the medical care process. Only then can we know how to do less at the bedside. We can and must distinguish between costly and high-quality care--they are not necessarily synonymous.


Changes in the National Prevalence of Current E-Cigarette, Cannabis, and Dual Use among Reproductive Age Women (18-44 Years Old) in the United States, 2013-2016.
Background: Electronic cigarette (e-cigarette) and cannabis (marijuana) use is rapidly increasing. Objectives: To report percentage prevalence and changes over time in current (past 30 days) e-cigarette, cannabis, and dual (concurrent) use in the population of reproductive age women (18-44 years old) in the United States. Methods: Our cross-sectional analysis involved data of 11, 004 women from Waves 1 to 3 of the Population Assessment of Tobacco and Health (PATH) Study (2013-2016). We estimated weighted percentage prevalence and 95% confidence intervals (CIs) and changes between 2013 and 2016 in current e-cigarette, cannabis, and dual use at each wave overall and across race/ethnicity, age, education, cigarette smoking status, alcohol use, and perceived mental health. Changes were reported as p for trend. Results: Between 2013 and 2016, e-cigarette use increased 13.6% (p for trend <.001) [15.2% (95% CI:14.2, 16.2) in 2013-2014; 22.2% (95% CI: 20.2, 24.3) in 2014-2015; and 28.8% (95% CI: 26.3, 31.3) in 2015-2016]; cannabis use increased 6.2% (p for trend <.001) [23.6% (95% CI: 22.1, 25.1) in 2013-2014; 28.5% (95% CI: 26.0, 31.0) in 2014-2015; and 29.8% (95% CI: 27.9, 31.1) in 2015-2016]; and dual use declined 0.3% (p for trend <.001) [5.8% (95% CI: 5.2, 6.3) in 2013-2014; 4.8% (95% CI: 3.7, 5.8) in 2014-2015; and 5.5% (95% CI: 4.2, 6.7) in 2015-2016]. Increases and declines in prevalence varied across sociodemographic characteristics, cigarette smoking status, alcohol use, and perceived mental health. Conclusions: Continued monitoring can capture further changes in prevalence and patterns to inform targeted public health intervention programs.


Durable Flexible Polymer-Encapsulated Cs4PbI6 Thin Film for High Sensitivity X-ray Detection.
Because of the extraordinary properties including high atomic numbers and large μτ products, metal halide perovskites have been widely employed and used for radiation detecting. Cs4PbI6 material has a high X-ray attenuation coefficient and excellent electrical properties that have a good potential in X-ray detection applications. Here, we have designed a flexible polymer-encapsulated Au/Cs4PbI6/Au X-ray detector with outstanding sensitivity of 256.20 μC Gy-1 cm-2 irradiated by 30 keV X-ray at 10 V bias, long-time stability, and durable flexibility without obvious degradation after bending for 600 cycles. These features demonstrate that this polymer-encapsulated durable flexible and sensitive X-ray detector could open a new possibility for next-generation radiation applications in dosimeter, imaging technologies.


Non-IgE Mediated Hypersensitivity to Food Products or Food Intolerance-Problems of Appropriate Diagnostics.
The diagnosis of food intolerance is tricky due to the different etiologies of adverse reactions. There is also a lack of clear rules for interpreting alternative tests used to diagnose these problems. The analyses of IgG4 concentration in serum or cytotoxic tests became the basis of elimination diets. However, it can result in nutritional deficiencies and loss of tolerance to eliminated foods. Our study aimed to assess the necessity of food elimination in four cases with food intolerance symptoms based on alternative diagnostic tests. Four patients without food allergies, who manifested diverse clinical symptoms after food, were presented due to the following factors: clinical history, diagnostic tests, elimination diet, and filaggrin gene (FLG) mutation. It was found that higher IgG4 levels against foods and higher cytotoxic test values are not clinically relevant in each of the studied individuals. They should not be decisive for the elimination of food products. The study of FLG-SNVs revealed the association of some clinical symptoms in patients with hypersensitivity to several food allergens and reported genetic variants in the FLG gene.


Demountable 3He melting curve thermometer.
We describe a demountable (3)He melting curve thermometer, suitable for use at low temperatures with liquid helium samples at any pressure. The thermometer's resolution is 10 microK below 0.l K. It is designed to be particularly useful for experiments performed in magnetic fields.


Systems Biology Modeling of the Complement System Under Immune Susceptible Pathogens.
The complement system is assembled from a network of proteins that function to bring about the first line of defense of the body against invading pathogens. However, complement deficiencies or invasive pathogens can hijack complement to subsequently increase susceptibility of the body to infections. Moreover, invasive pathogens are increasingly becoming resistant to the currently available therapies. Hence, it is important to gain insights into the highly dynamic interaction between complement and invading microbes in the frontlines of immunity. Here, we developed a mathematical model of the complement system composed of 670 ordinary differential equations with 328 kinetic parameters, which describes all three complement pathways (alternative, classical, and lectin) and includes description of mannose-binding lectin, collectins, ficolins, factor H-related proteins, immunoglobulin M, and pentraxins. Additionally, we incorporate two pathogens: (type 1) complement susceptible pathogen and (type 2) Neisseria meningitidis located in either nasopharynx or bloodstream. In both cases, we generate time profiles of the pathogen surface occupied by complement components and the membrane attack complex (MAC). Our model shows both pathogen types in bloodstream are saturated by complement proteins, whereas MACs occupy <<1.0% of the pathogen surface. Conversely, the MAC production in nasopharynx occupies about 1.5-10% of the total N. meningitidis surface, thus making nasal MAC levels at least about eight orders of magnitude higher. Altogether, we predict complement-imbalance, favoring overactivation, is associated with nasopharynx homeostasis. Conversely, orientating toward complement-balance may cause disruption to the nasopharynx homeostasis. Thus, for sporadic meningococcal disease, our model predicts rising nasal levels of complement regulators as early infection biomarkers.


DNA methylation and leukemia susceptibility in China: Evidence from an updated meta-analysis.
Mounting evidence supports a role for DNA methylation in the pathogenesis of leukemia; however, there no overview of these results in the Chinese population. The present study performed a comprehensive meta-analysis to establish candidate genes with an altered methylation status in Chinese leukemia patients. Eligible studies were identified through searching the National Center of Biotechnology Information PubMed and Wanfang databases. Studies were pooled and overall odds ratios with corresponding confidence intervals were calculated. A total of 4,325 leukemia patients and 2,010 controls from 94 studies on 53 genes were included in this meta-analysis, and 47 genes were found to be aberrantly methylated in leukemia patients. A further subgroup meta-analysis by leukemia subtype demonstrated that hypermethylation of 5 genes, namely cyclin-dependent kinase (CDKN)2A, DNA-binding protein inhibitor-4, CDKN2B, glioma pathogenesis-related protein 1 and p73, contributed to the risk of various subtypes of leukemia. In addition, a strong association between CDKN2A and leukemia was identified in Chinese (P<0.00001) but not in European patients. The aberrantly methylated genes identified in the present meta-analysis may help elucidate the mechanisms underlying the development of leukemia in Chinese patients.


Comparison between the BACTEC 9240 and the Pall eBDS system for detection of bacterial platelet concentrate contamination.
Only two commercially available automated systems have been cleared by the FDA for screening of bacterial contamination in platelet (PLT) products. These are the Pall eBDS (Pall Corp.), based on measurement of oxygen consumption by contaminant organisms, and the BacT/ALERT (bioMérieux), revealing increasing carbon dioxide concentration due to bacterial growth. The authors compared the performance of the Pall eBDS with the BACTEC 9240 (bioMérieux) in detecting PLT contamination. Serial dilutions of 10 bacterial species frequently associated with PLT contamination were prepared in an apheresis PLT unit per organism. Units were from single donors. After 30 minutes from seeding PLT units, a volume of suspension achieving a final bacterial concentration of 1 to 10 colony-forming units/mL for each unit was inoculated in two Pall bags and a BACTEC bottle, and the same was done after 24 hours from seeding. Measurements were performed at 24 and 30 hours. Significant differences between the two instruments were only found when screening PLT units after 24 hours from seeding. The Pall system showed a higher sensitivity than BACTEC 9240, because it revealed 97 and 98% of positive samples at 24 and 30 hours of incubation, respectively, whereas the second detected 86 and 90% of contaminated products. Significance was lost after 35-hour incubation with the BACTEC 9240. By comparing the two instruments, their performances were found to be comparable; the Pall system appeared as a more suitable method when using 24 to 30 hours as times for readings, but the significant difference was lost after 35-hour incubation.


Prognostic Significance of Low HER2 Expression in Patients With Metastatic Hormone Receptor-positive Breast Cancer Treated With First Line CDK4/6 Inhibitors: A Greek Multicenter Real-world Data Analysis.
Low expression of HER2 has defined a new "HER2-low" subgroup of breast cancer with distinct clinicopathological characteristics and both prognostic and predictive implications. The impact of low HER2 expression in metastatic hormone receptor-positive HER2-negative breast cancer treated with first-line CDK4/6 inhibitors has not been studied. Using real-world patient data, we aimed to identify prognostic differences in this patient population according to HER2 expression with immunohistochemistry. We retrospectively analyzed 191 patients from 5 Oncology Department databases in Thessaloniki, Greece, with hormone receptor-positive HER2-negative metastatic breast cancer treated with CDK4/6 inhibitors in the first line, for whom detailed immunohistochemical HER2 data could be retrieved. Median progression-free survival was numerically different among the different HER2 subgroups (3.35 years for HER2 0 tumors, 2.18 years for HER2 +1 tumors, 1.74 years for HER2 +2/ISH-negative tumors), but this difference was not statistically significant (p=0.477). Median PFS was statistically significantly longer in patients without visceral metastases (5.45 years) compared to patients with visceral metastases (1.61 years) (p=0.017). Median PFS was also statistically significantly longer in patients taking an aromatase inhibitor (2.99 years) compared to patients taking fulvestrant (1.33 years) (p<0.0001). There were no statistically significant differences in the other subgroups examined. CDK4/6 inhibitors are equally effective as first-line treatment agents, regardless of the exact level of HER2 expression. Numerical differences, however, do exist among the different HER2 subgroups, and merit further evaluation in future studies to better study this phenomenon.


A case report of acute myeloid leukemia and neurofibromatosis 1.
We report a case of a 65-year old patient affected by neurofibromatosis 1, documented by the presence of germ-line mutation on the NF1 gene, who developed various hyperproliferative malignant and benign diseases. He was brought to our attention for the diagnosis of acute myeloid leukemia revealed by major fatigue and dyspnea. The disease characteristics at diagnosis were hyperleukocytosis and complex karyotype with the inversion of the chromosome 16, classifying as a high-risk leukemia. The association between leukemia and neurofibromatosis 1 is controversial and needs to be further investigated. Nevertheless, such patients present a wide number of comorbidities that make therapeutic strategies most difficult.


Investigation of factors from assistive technology professionals that impact timeliness of wheelchair service delivery: a cross-sectional study.
The purpose of this study was to examine factors associated with variability in time from assessment to device delivery (cycle time). Our hypothesis was that device type and type of insurance would be the strongest predictor of cycle time. Data were extracted from the Functional Mobility Assessment/Uniform Dataset (FMA/UDS) Registry that at the time of analysis contained a sample of 2588 people with disabilities (PWD) who were provided with a wheeled mobility device (WMD) between 21 March 2016 and 29 June 2021. To examine the effect of individual factors on the variability in cycle time, a robust linear regression analysis was conducted. The average national cycle time was 101.5 (SD = 59.9) d. Geographic area (Capital Metro [p < .001], Great Lakes [p = .016], and Northeast area [p < .001]), higher years since onset of disability (p < .001) and customizable devices (p = .021) were associated with higher cycle time. Non-customizable devices (p = .005), scooters (p < .001), Group 2 power wheelchairs (PWCs; p < .001), and funding source (Medicaid managed care (p < .001) and "other" (p = .028)) were associated with lower cycle time. Longer cycle time is likely related to variations in clinical practice, insurance coverage criteria and the level of customizability of the device needed for a particular diagnosis, especially long-term disabilities.Implications for rehabilitationThe national average number of days between initial evaluation and device delivery (cycle time) to deliver a wheeled mobility device (WMD) varies based on specific variables such as type of WMD, diagnosis and payer source.Geographic area, years since onset of disability, device type, primary diagnosis and funding source significantly impact cycle times.Increased complexity of the WMD, both manual and power wheelchairs (PWCs), was associated with longer cycle times.As more service delivery models emerge, specific benefits and challenges need to be reported on how they impact cycle time.


Hydrazide and hydrazine reagents as reactive matrices for matrix-assisted laser desorption/ionization mass spectrometry to detect steroids with carbonyl groups.
Fifty-one reagents (37 hydrazides and 14 hydrazines) were examined to determine if they could function as reactive matrices for the detection of steroids with carbonyl groups in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Among these reagents, 3-hydroxy-2-naphthoic acid hydrazide reacted most efficiently with steroids with carbonyl groups, allowing detection of them at the 1pmol level. Results from accurate mass measurement with a JMS-S3000 Spiral-TOF suggested that the ion peaks corresponding to the protonated fragment of a hydrazone were observed, in addition to the protonated molecular peaks of the resulting hydrazones ([M(D)+H](+)) and their sodium adduct ([M(D)+Na](+)). 3-Hydroxy-2-naphthoic acid hydrazide also could detect cortisol (1.93ng) spiked in a human serum certified reference material, as well as cortisone and cortisol in a human saliva sample collected in the morning. Using a combination of 3-hydroxy-2-naphthoic acid hydrazide and α- cyano-4-hydroxycinammic acid (CHCA) (3-hydroxy-2-naphthoic acid hydrazide:CHCA=4:1, v/v), even cortisone and cortisol in a human saliva sample collected in the afternoon, when levels are typically low, could be detected effectively.


Peroxisome proliferator-activated receptor-γ and its related pathway in bone marrow mesenchymal stem cell differentiation co-cultured with mechanically stretched ligament fibroblasts.
The occurrence of pelvic floor dysfunctional disease (PFD) is closely related with elasticity, toughness, and functional changes of the connective tissue of the pelvic support tissue. Bone marrow mesenchymal stem cells (BMSCs) have been confirmed to have the capacity to differentiate into a variety of cell types such as osteoblasts, chondroblasts, adipocytes and fibroblasts. Therefore, BMSCs have the potential to improve the clinical outcomes for PFD. Peroxisome proliferator-activated receptor-γ (PPAR-γ), a ligand activated transcription factor, has acquired a great deal of attention as it is involved in the fibrosis and cell differentiation. However, how it is regulated during the process of the differentiation of BMSCs into fibroblasts remains to be defined. The present study investigated the underlying mechanisms of PPAR-γ effect of mechanical stretch on the differentiation of BMSCs induced by pelvic ligament fibroblasts. PPAR-γ expression was decreased during the differentiation of BMSCs into fibroblasts by co-cultured stretched fibroblasts. Addition of transforming growth factor-β1 (TGF-β1) reduced PPAR-γ expression and promoted the differentiation of BMSCs. With the employment of endogenous ligand, activation of PPAR-γ suppressed the BMSC differentiation. Similar effects were also observed with overexpression of PPAR-γ gene. In addition, decrease of PPAR-γ by the use of shRNA targeting rat PPAR-γ significantly contributed to BMSC differentiation to fibroblasts. These results indicate that PPAR-γ negatively regulates the differentiation of BMSCs into fibroblasts.


Imaging alternatives to colonoscopy: CT colonography and colon capsule. European Society of Gastrointestinal Endoscopy (ESGE) and European Society of Gastrointestinal and Abdominal Radiology (ESGAR) Guideline - Update 2020.
1. ESGE/ESGAR recommend computed tomographic colonography (CTC) as the radiological examination of choice for the diagnosis of colorectal neoplasia. Strong recommendation, high quality evidence. ESGE/ESGAR do not recommend barium enema in this setting. Strong recommendation, high quality evidence.2. ESGE/ESGAR recommend CTC, preferably the same or next day, if colonoscopy is incomplete. The timing depends on an interdisciplinary decision including endoscopic and radiological factors. Strong recommendation, low quality evidence. ESGE/ESGAR suggests that, in centers with expertise in and availability of colon capsule endoscopy (CCE), CCE preferably the same or the next day may be considered if colonoscopy is incomplete. Weak recommendation, low quality evidence.3. When colonoscopy is contraindicated or not possible, ESGE/ESGAR recommend CTC as an acceptable and equally sensitive alternative for patients with alarm symptoms. Strong recommendation, high quality evidence. Because of lack of direct evidence, ESGE/ESGAR do not recommend CCE in this situation. Very low quality evidence. ESGE/ESGAR recommend CTC as an acceptable alternative to colonoscopy for patients with non-alarm symptoms. Strong recommendation, high quality evidence. In centers with availability, ESGE/ESGAR suggests that CCE may be considered in patients with non-alarm symptoms. Weak recommendation, low quality evidence.4. Where there is no organized fecal immunochemical test (FIT)-based population colorectal screening program, ESGE/ESGAR recommend CTC as an option for colorectal cancer screening, providing the screenee is adequately informed about test characteristics, benefits, and risks, and depending on local service- and patient-related factors. Strong recommendation, high quality evidence. ESGE/ESGAR do not suggest CCE as a first-line screening test for colorectal cancer. Weak recommendation, low quality evidence.5. ESGE/ESGAR recommend CTC in the case of a positive fecal occult blood test (FOBT) or FIT with incomplete or unfeasible colonoscopy, within organized population screening programs. Strong recommendation, moderate quality evidence. ESGE/ESGAR also suggest the use of CCE in this setting based on availability. Weak recommendation, moderate quality evidence.6. ESGE/ESGAR suggest CTC with intravenous contrast medium injection for surveillance after curative-intent resection of colorectal cancer only in patients in whom colonoscopy is contraindicated or unfeasible. Weak recommendation, low quality evidence. There is insufficient evidence to recommend CCE in this setting. Very low quality evidence.7. ESGE/ESGAR suggest CTC in patients with high risk polyps undergoing surveillance after polypectomy only when colonoscopy is unfeasible. Weak recommendation, low quality evidence. There is insufficient evidence to recommend CCE in post-polypectomy surveillance. Very low quality evidence.8. ESGE/ESGAR recommend against CTC in patients with acute colonic inflammation and in those who have recently undergone colorectal surgery, pending a multidisciplinary evaluation. Strong recommendation, low quality evidence.9. ESGE/ESGAR recommend referral for endoscopic polypectomy in patients with at least one polyp ≥6 mm detected at CTC or CCE. Follow-up CTC may be clinically considered for 6-9-mm CTC-detected lesions if patients do not undergo polypectomy because of patient choice, comorbidity, and/or low risk profile for advanced neoplasia. Strong recommendation, moderate quality evidence. Source and scope This is an update of the 2014-15 Guideline of the European Society of Gastrointestinal Endoscopy (ESGE) and the European Society of Gastrointestinal and Abdominal Radiology (ESGAR). It addresses the clinical indications for the use of imaging alternatives to standard colonoscopy. A targeted literature search was performed to evaluate the evidence supporting the use of computed tomographic colonography (CTC) or colon capsule endoscopy (CCE). The Grading of Recommendations Assessment, Development and Evaluation (GRADE) system was adopted to define the strength of recommendations and the quality of evidence.


Methods for detecting disease clustering, with consideration of childhood leukaemia.
In trying to interpret reports of disease clusters in specific localities, it is valuable to know whether the disease in question has a general tendency to cluster spatially. Methods for investigating localized disease clustering were the subject of a comparative study organized by the International Agency for Research on Cancer some years ago. This paper addresses some further aspects of one of the methods used in this exercise, namely the Potthoff-Whittinghill (P-W) test. Particular consideration is given to methodology for estimating the magnitude of overdispersion and for detecting whether one area, in particular, has an undue influence on the evidence for overdispersion using the P-W test, the extent to which is possible to detect clustering over regions of differing sizes using a components-of-variance approach and how adjustment for overdispersion might affect tests for raised disease rates in specific locations. These points are illustrated using data on childhood leukaemia incidence and reference is made to other analyses of the geographical distribution of childhood leukaemia that are based on this approach.


Electrically templated dewetting of a UV-curable prepolymer film for the fabrication of a concave microlens array with well-defined curvature.
This paper presents an economic method, based on electrically templated dewetting of a UV-curable prepolymer, for fabricating a concave microlens array (MLA) of high quality and high density. In our strategy, a voltage is applied to an electrode pair consisting of a conductive substrate coated with a UV-curable prepolymer film and a microhole-arrayed silicon template, sandwiching an air gap, to dewet the prepolymer film into a curved air-liquid interface. At or beyond a critical voltage, the curved prepolymer can be pulled quickly into contact with the protrusive underside of the silicon template. Contact of the prepolymer with the template can be detected by monitoring the leaky current in the polymer, followed by a UV curing of the prepolymer. Finally, by separating the mold from the solidified polymer, a concave MLA is obtained. The curvature of the MLA can be well-defined simply by changing the air gap between the mold and prepolymer film. Besides, the dewetting strategy results in a much smaller adhesion area between the mold and solidified polymer structures, which allows for easy separation of the mold from the MLA in a large-area operation.


Polythiophene-fullerene based photodetectors: tuning of spectral response and application in photoluminescence based (bio)chemical sensors.
A photoluminescence (PL)-based oxygen and glucose sensor utilizing inorganic or organic light emitting diode as the light source, and polythiophene: fullerene type bulk-heterojunction devices as photodetectors, for both intensity and decay-time based monitoring of the sensing element's PL. The sensing element is based on the oxygen-sensitive dye Pt-octaethylporphyrin embedded in a polystyrene matrix.


The synthesis and radiolabeling of 2-nitroimidazole derivatives of cyclam and their preclinical evaluation as positive markers of tumor hypoxia.
The cyclam ligand (1,4,8,11-tetraazacyclotetradecane) was condensed with various azomycin-containing synthons to produce chemical compounds that could chelate radioactive metals. It was expected that these radiolabeled markers would become bound selectively to hypoxic cells on the bioreduction of their azomycin substituent. The markers were radiolabeled with (99m)Tc, (67)Cu, or (64)Cu. Their uptake and binding to tumor cells in vitro was characterized as a function of time and oxygen concentration. These data defined the hypoxia-specific factor, the ratio of the initial rate of marker binding to severely hypoxic relative to aerobic cells. In addition, the concentration of oxygen (in the equilibrium gas phase) that inhibited binding to 50% of the maximum rate was determined. The in vivo biodistribution and clearance kinetics of the favorable markers were investigated with severe combined immune deficiency mice bearing EMT-6 tumors whose radiobiologic hypoxic fraction (RHF) was approximately 40%. The specific activity (percentage injected dose per gram [%ID/g]) in normal and tumor tissue and the tumor-to-blood and tumor-to-muscle ratios of the optimal markers were also measured for Dunning prostate carcinomas of anaplastic (RHF = 15%-20%) and well-differentiated (RHF < 1%) histology growing in Fischer X Copenhagen rats. Planar images were acquired with some markers from these tumor-bearing rats. The tumor uptake of these cyclam-based markers is approximately 10 times higher when they are labeled with copper isotopes than when labeled with (99m)Tc. FC-327 and FC-334, di-azomycin-substituted cyclams, exhibited hypoxia-specific factors > or = 7.0. The oxygen concentration that inhibited their binding to 50% of the maximal rate was approximately 0.5% O(2), similar to that of the radiobiologic oxygen effect. The %ID/g of (64)Cu-FC-334 retained in EMT-6 tumors in mice and in the anaplastic and well-differentiated prostate tumors in rats 6 h after administration was approximately 6.5, 0.4, and 0.1, respectively. Marker activity in tumor was always less than that in liver and kidney. The tumor-to-blood and tumor-to-muscle ratios of (64)Cu-FC-327 and (64)Cu-FC-334 activity in R3327-AT tumor-bearing rats are higher than those observed for (64)Cu-di-acetyl-bis (N(4)-methylthiosemicarbazone) and approach those of beta-D-(125)I-iodinated azomycin galactopyranoside, the optimal hypoxia marker of the azomycin-nucleoside class. These data suggest that some azomycin-cyclams exhibit good hypoxia-marking potential to tumor cells in vitro and to animal tumors of known RHF. Both PET and SPECT could be used to image tumor hypoxia with markers labeled with (64)Cu and (67)Cu, respectively.


A new fibroblast growth stimulating activity from the human megakaryoblastic leukaemia cell line ELF-153: in vitro and in vivo findings.
Although the exact mechanism for the progression of myelofibrosis in acute megakaryoblastic leukaemia is unclear, certain humoral factors released from the proliferating megakaryoblasts that are unable to store these factors in their defective alpha-granules, including platelet derived growth factor (PDGF), fibroblast growth factors (FGF), platelet factor-4 (PF-4), transforming growth factor-beta (TGF-beta) and beta-thromboglobulin, could result in increased collagen synthesis by bone marrow fibroblasts. Recently, the human megakaryoblastic leukaemia cell line MEG-01 has been shown to produce both TGF-beta and PF-4 which have enhanced the growth of bone marrow fibroblasts. Therefore, we have examined the presence of a fibroblast growth stimulating activity and the humoral factors that might be responsible for it in the supernatant of the human megakaryoblastic leukaemia cell line ELF-153 recently established in our laboratory from a patient with acute myelofibrosis. A new fibroblast growth stimulating activity has been identified in the supernatant of the ELF-153 human megakaryoblastic leukaemia cell line that is independent of the percentage of fetal calf serum in NRK-49F fibroblast agar clonogenic assays and is not due to any of the known fibroblast growth stimulating humoral factors including PDGF, epithelial growth factor, TGF-alpha or beta, tumour necrosis factor-alpha, interleukin-1, 2, 4 or 6, FGF, fibronectin, PF-4 and factor VIII AG. Also, in vivo, subcutaneous injection of ELF-153 megakaryoblastic leukaemia cells into nude mice formed, in three out of the five mice after 6 weeks, subcutaneous tumours with a very rigid texture whose histological examination revealed dense infiltration by blast cells and pronounced reticular fibrosis. Immunohistochemistry demonstrated exclusive deposition of collagen III in the extracellular matrix whereas laminin and collagen IV were absent.(ABSTRACT TRUNCATED AT 250 WORDS)


C-terminal ladder sequencing via matrix-assisted laser desorption mass spectrometry coupled with carboxypeptidase Y time-dependent and concentration-dependent digestions.
The utility of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry for the analysis of C-terminal peptide ladders from carboxypeptidase Y (CPY) digestions is discussed. MALDI analysis of aliquots of an optimized time-dependent CPY digestion of ACTH 7-38 fragment allowed for the sequence of the first 19 amino acids from the C-terminus to be determined in 25 min of digestion time. A strategy for performing parallel concentration-dependent digestions on the MAL-DI plate is proven to be superior to the time-dependent approach as the method development time and practical amounts of both peptide and enzyme consumed are reduced significantly. The on-plate approach offered the same sequence information from the ACTH 7-38 fragment and was used to digest 22 peptides of various amino acid composition, size, charge, and polarity. Of the 22 peptides digested on-plate, sequence information was derived from 19 of them. A statistical analysis strategy for ladder sequencing utilizing t-statistics is offered as a method for placing confidence intervals on residue assignments.


Nosocomial infections in burned patients in motahari hospital, tehran, iran.
Burn patients are at high risk of developing nosocomial infection because of their destroyed skin barrier and suppressed immune system, compounded by prolonged hospitalization and invasive therapeutic and diagnostic procedures. Studies on nosocomial infection in burn patients are not well described. The objective of the present study was to identify the causative bacterial of nosocomial infection and to determine the incidence of nosocomial infection and their changing during hospitalization in burned patients admitted to in the Motahari Hospital, Tehran, Iran. During the second part of 2010, 164 patients were included in this study. Samples were taken the first 48 hours and the fourth week after admission to Motahari Burn hospital. Isolation and identification of microorganisms was performed using the standard procedure. Of the 164 patients, 717 samples were taken and 812 bacteria were identified, 610 patients were culture positive on day 7 while 24 (17.2%) on 14 days after admission. The bacteria causing infections were 325 Pseudomonas, 140 Acinetobacter, 132 Staphylococcus aureus, and 215 others. The percentage of mortality was 12%. All of patients had at least 1 positive culture with Pseudomonas and/or with Acinetobacter. Hospitals suggest continuous observationof burn infections and increase strategies for antimicrobial resistance control and treatment of infectious complications.


A case of metastatic adrenocortical carcinoma diagnosed with steroidogenic factor-1 in a sprague-dawley rat.
This report describes the morphological and immunohistochemical characteristics of an adrenocortical carcinoma with distant metastasis in a Sprague-Dawley rat. Macroscopically, a single large mass was observed in the adrenal gland, and multiple nodules were noted in the lung, liver and thyroid. Histologically, the adrenal tumor consisted of a solid growth of eosinophilic round cells with nuclear atypia. Vascular invasion was present, and multiple metastatic lesions were also observed in the lungs, liver, and mediastinal lymph nodes. Immunohistochemically, the nuclei of these tumor cells were positive for Steroidogenic Factor-1 (SF-1). In the thyroid, tumor cells histologically resembling adrenal cells were immunohistochemically negative for SF-1 but positive for calcitonin; thus the lesion was diagnosed as thyroid C-cell carcinoma. From these results, the present case was diagnosed as adrenocortical carcinoma with distant metastases. SF-1 could be a valuable marker for the differential diagnosis of adrenocortical tumors versus other endocrine tumors such as C-cell carcinoma.


Gastropericardial fistula and Candida kruzei pericarditis following laparoscopic Nissen fundoplication (gastropericardial fistula).
We report a case of gastropericardial fistula and Candida kruzei pericarditis one year after laparoscopic Nissen fundoplication. Chest X-ray and CT revealed pneumopericardium. Barium swallow, UGI, endoscopy, and bronchoscopy were negative. Pericardial exploration was performed through a sternotomy. Intraoperative fistulogram revealed a tract into the stomach. A midline abdominal incision allowed identification of the slipped Nissen, resection of the fistula tract, and subsequent re-do fundoplication. The patient was treated with amphotericin and remains symptom-free 10 months after her operation. We recommend both sternotomy and midline abdominal incisions to explore and access the pericardium, stomach, esophagus, and diaphragm.


Psychiatric implications of hepatitis-C infection.
Hepatitis-C virus (HCV) has infected an estimated 130 million people worldwide, most of whom are chronically infected. Infection is marked by both treatment- and non-treatment-related psychiatric symptoms. Symptoms associated with antiretroviral therapy, interferon-alpha (IFN-alpha), include acute confusional states, delirium, depression, irritability, and even mania. These psychiatric symptoms are further complicated by the high rate of substance abuse and comorbid HIV infection inherent to this population. Even in the absence of IFN-alpha therapy, comorbid depression, cognitive decline, and especially fatigue are common in patients suffering HCV. These comorbidities have significant effects on both treatments and outcomes, and thus are reviewed herein.


Uncertainty in least-squares fits to the thermal noise spectra of nanomechanical resonators with applications to the atomic force microscope.
Thermal noise spectra of nanomechanical resonators are used widely to characterize their physical properties. These spectra typically exhibit a Lorentzian response, with additional white noise due to extraneous processes. Least-squares fits of these measurements enable extraction of key parameters of the resonator, including its resonant frequency, quality factor, and stiffness. Here, we present general formulas for the uncertainties in these fit parameters due to sampling noise inherent in all thermal noise spectra. Good agreement with Monte Carlo simulation of synthetic data and measurements of an Atomic Force Microscope (AFM) cantilever is demonstrated. These formulas enable robust interpretation of thermal noise spectra measurements commonly performed in the AFM and adaptive control of fitting procedures with specified tolerances.