Characterization of a soluble KGF receptor cDNA from human corneal and breast epithelial cells.
Keratinocyte growth factor (KGF) is a member of the fibroblast growth factor (FGF) family FGF-7. It exhibits potent mitogenic activity for epithelial cells, including corneal and mammary epithelial cells. A messenger RNA has been reported that is generated by alternative splicing of bek that putatively codes only for the extracellular ligand-binding domain of KGF receptor (soluble KGF receptor). In the present study, the expression of the mRNA coding for this alternative bek transcript was examined and the corresponding protein characterized. Alternative messenger RNA transcripts were detected in various cell lines or tissues using reverse transcription-polymerase chain reaction (RT-PCR) and RNase protection assay. NIH/3T3 fibroblast cells and 293 kidney embryonic epithelial cells were stably transfected with soluble KGF receptor cDNA and transmembrane KGF receptor cDNA. Soluble KGF receptor protein was produced using a baculovirus-insect expression system. Soluble KGF receptor protein was detected using western and dot blot analyses. Binding assays and cross-linking labeling were used to determine the affinity and specificity of soluble KGF receptor. A mitogenic assay was performed to examine the function of the soluble KGF receptor. The soluble KGF receptor mRNA was primarily expressed in epithelial cells, including cells from the cornea and breast. Cross-linking labeling and affinity-binding assays with 125I-KGF showed that the soluble KGF receptor bound KGF (FGF-7) but not FGF-1 or FGF-2. Soluble KGF receptor was detected in the culture medium of cells stably transfected with soluble KGF receptor cDNA but not with transmembrane KGF receptor cDNA, suggesting that the soluble receptor was generated by mRNA splicing and probably not by proteolysis or posttranslational processing. Soluble KGF receptor inhibited KGF binding to transmembrane KGF receptor and DNA synthesis in BALB/MK epidermal keratinocytes in response to KGF, suggesting that soluble KGF receptor expression could provide a mechanism for the cell to downregulate responses to KGF. A truncated soluble KGF receptor expressed in corneal and other epithelial cells probably functions to downregulate the response of the cell to KGF.


In this case report, a 61-year-old male presented with odynophagia and ulceration in palatum durum after inhalating dust from machinery containing a weak acid. It was at first diagnosed as an acidic ulcer due to two biopsies verifying this. Because of progressing ulceration a third biopsy was taken - this time with the diagnosis extranodal NK/T-cell lymphoma, nasal type. This illustrates the diagnostic challenges of the illness, typically requiring multiple biopsies, and one should have this differential diagnosis in mind in case of progressing ulceration.


The Online Art Museum.
This article was migrated. The article was marked as recommended. With the onset of the coronavirus-19 (COVID-19) pandemic, we transformed an in-person art museum-based course for medical students into an online format. This brought new challenges but offered unexpected advantages. The course included daily close-looking of artworks using the Visual Thinking Strategies method, group arts-based activities, reflective writing, and independent creating assignments. The virtual format allowed us to incorporate important features that were unavailable in our in-person elective: multi-media activities, access to nearly unlimited international works of art, and personal reflection from one's private space. As instructors, the experience enlightened us on the value of online arts-based teaching.


Trauma-induced central diabetes insipidus in a cat.
A 1-year-old neutered male domestic shorthair cat presented with a 4-week history of polydipsia that began immediately after an 8 m fall. Trauma-induced central diabetes insipidus was suspected on the basis of the identification of hyposthenuria, normal haematology and serum biochemistry profile and unremarkable abdominal ultrasound examination. Failure to concentrate urine with water deprivation followed by production of hypersthenuric urine with administration of the synthetic antidiuretic hormone, Damino-8-D-arginine vasopressin (DDAVP), confirmed the diagnosis of central diabetes insipidus. Treatment via conjunctival administration of DDAVP failed to attenuate the polydipsia, however, resolution of polydipsia was achieved with subcutaneous administration of DDAVP and the cat remains eudipsic with twice daily subcutaneous DDAVP administration 17 months after diagnosis.


Neuroendocrine tumor of the common bile duct: case report.
Neuroendocrine tumors (NET) are a very heterogeneous group of neoplasms; in recent years we have seen an increase in their incidence (3.65 /100.000/year). They can be associated with hereditary endocrine syndromes (MEN, Von Hippel Lindau); they can occur at any age and the incidence is slightly higher in men than women. The aetiology of the neuroendocrine tumors is unclear; in most cases, inflammation of the bile ducts may be the underlying cause and for this reason, the initial patient's evaluation should be focused on the different aspects concerning the oncological one and the possible sequelae of the biliary obstructions that can evolve in biliary sepsis. All neuroendocrine tumors have malignant potential. The most frequent sites of extrahepatic biliary NETs are the common hepatic duct and the distal common bile duct (19.2%), followed by the middle of the common bile duct (17.9%), the cystic duct (16.7%), and the proximal common bile duct (11.5%). We can divide them into: well-differentiated and poorly differentiated. Considering the clinical features, neuroendocrine tumors can be divided into functional and non-functional. As regards the staging, we distinguish localized, regional and metastatic tumors. Tumors derived from the bile duct are difficult to diagnose preoperatively, mainly because of its low incidence and difficult diagnostic process. However since cholangiocarcinomas account for about 80% of all primary biliary tumors, it is important to think about other options despite their low frequency when a patient presents with abnormal characteristics. The most sensitive immunohistochemical markers are expressing neuron-specific enolase, synaptophisin and chromogranin A. Liver function tests, alkaline phosphatase and bilirubin are often high. Sometimes an anemia can appear in the presence of a chronic disease or in patients with more advanced disease. It is known that the measurement of chromogranin A is useful for the preoperative diagnosis of neuroendocrine tumors. Chromogranin A is elevated in 90% of neuroendocrine tumors of the intestine, and the levels correlate with tumor burden and the possibility of recurrence and, therefore, chromogranin A can be an effective biological marker for preoperative diagnosis of neuroendocrine tumors. Bile endocrine tumors remain silent until metastasizing or growing into neighboring organs, because of its uncommon diagnosis in early stages due to its low incidence, absence of serum markers and lack of symptoms related to the hormonal pattern. Preoperative diagnosis of common bile duct carcinoma is extremely difficult, because it is foretold by non-specific symptoms that include pain or discomfort in the right upper quadrant level and weight loss. A 51- year-old woman presented a jaundice and severe bile duct dilatation. The enhanced CT scan showed a mass, approximately 15 mm in diameter, in the distal common biliary duct. The MRI and ERCP confirmed the mass. Cromogranin A value was negative. The diagnosis of well differentiated endocrine tumor of the biliary tract was done after its surgical resection was performed. The postoperative period was uneventful. Extrahepatic biliary NETs are rare, and extrahepatic bile ducts reportedly account for only 0.32% of primary NET sites. The prognosis for NET of the bile duct appears to be poor.


In Vitro Safety "Clinical Trial" of the Cardiac Liability of Hydroxychloroquine and Azithromycin as COVID19 Polytherapy.
Despite global efforts, there are no effective FDA-approved medicines for the treatment of SARS-CoV-2 infection. Potential therapeutics focus on repurposed drugs, some with cardiac liabilities. Here we report on a preclinical drug screening platform, a cardiac microphysiological system (MPS), to assess cardiotoxicity associated with hydroxychloroquine (HCQ) and azithromycin (AZM) polytherapy in a mock clinical trial. The MPS contained human heart muscle derived from patient-specific induced pluripotent stem cells. The effect of drug response was measured using outputs that correlate with clinical measurements such as QT interval (action potential duration) and drug-biomarker pairing. Chronic exposure to HCQ alone elicited early afterdepolarizations (EADs) and increased QT interval from day 6 onwards. AZM alone elicited an increase in QT interval from day 7 onwards and arrhythmias were observed at days 8 and 10. Monotherapy results closely mimicked clinical trial outcomes. Upon chronic exposure to HCQ and AZM polytherapy, we observed an increase in QT interval on days 4-8.. Interestingly, a decrease in arrhythmias and instabilities was observed in polytherapy relative to monotherapy, in concordance with published clinical trials. Furthermore, biomarkers, most of them measurable in patients' serum, were identified for negative effects of single drug or polytherapy on tissue contractile function, morphology, and antioxidant protection. The cardiac MPS can predict clinical arrhythmias associated with QT prolongation and rhythm instabilities. This high content system can help clinicians design their trials, rapidly project cardiac outcomes, and define new monitoring biomarkers to accelerate access of patients to safe COVID-19 therapeutics.


Characterization of glucocorticoid receptors and glucocorticoid receptor mRNA in human leukemia cells: stabilization of the receptor by diisopropylfluorophosphate.
We have shown that cytosol samples from human leukemia cells frequently contain glucocorticoid receptor fragments that have a mol wt (Mr) of approximately 52,000. In the present study we demonstrate that the Mr approximately 52,000-receptor fragments are derived from intact glucocorticoid receptors (Mr approximately 97,000) by the action of a serine protease. Mr approximately 52,000-receptor fragments were present in cytosol from 24 of 52 leukemia cell samples. Only normal size glucocorticoid receptors were present in cytosol samples if diisopropylfluorophosphate (DFP), a potent inhibitor of serine proteases, was added to the hypotonic buffer used for cytosol preparation. Receptor proteolysis was not inhibited by hydrolyzed DFP, benzamidine, phenylmethylsulfonylfluoride, aprotinin, iodoacetamide, or mercuric chloride. The leukemia cell protease digests the receptor at a different site than chymotrypsin, which digests the intact receptor to produce a Mr approximately 40,000 receptor fragment. Receptor messenger RNA (mRNA) in S49 mouse lymphoma cells and in human leukemia cells was analyzed by Northern hybridization with a cDNA for the normal glucocorticoid receptor. Mutant S49 mouse lymphoma cells that have abnormally small glucocorticoid receptors (Mr approximately 48,000) make a 5.0-kilobase receptor transcript in addition to the normal size 6.5-kilobase receptor transcript. A normal size receptor transcript of 6.5 kilobases was present in all of the human leukemia cells whether or not Mr approximately 52,000-receptor fragments were present. Therefore, abnormalities of glucocorticoid receptor mRNA, which may give rise to the synthesis of foreshortened receptors in certain mutant mouse lymphoma cells, are apparently absent from human leukemia cells.


Monocarboxylate transporter 1 mediates biotin uptake in human peripheral blood mononuclear cells.
Here the hypothesis was tested that monocarboxylate transporters (MCT) mediate biotin transport in human lymphoid cells. Uptake of [(3)H]biotin was measured in human lymphoid cells [peripheral blood mononuclear cells (PBMC) and Jurkat cells] under conditions known to affect MCT-mediated transport. When biotin uptake into PBMC was measured in the presence of excess concentrations of competitors (MCT substrates) and MCT inhibitors, transport rates decreased significantly to <75 and <67%, respectively, of controls. Biotin uptake correlated with the concentration of protons in culture media, consistent with cotransport of protons and the carboxylate biotin by MCT. Efflux of biotin from PBMC was stimulated by extracellular lactate (a known substrate for MCT), consistent with countertransport of the two substrates by the same transporter. PBMC responded to proliferation with parallel increases of transport rates for both biotin and lactate, providing circumstantial evidence that the same transporter mediates uptake of the two substrates in PBMC. Transfection of Jurkat cells with an expression vector encoding MCT1 caused a 50% increase in biotin uptake; in contrast, overexpression of MCT1 did not affect biotin uptake in various nonlymphoid cell lines. These findings are consistent with the hypothesis that MCT mediate biotin uptake in human lymphoid cells.


Collagenase production by human mononuclear cells in culture: inhibition by gold containing compounds and other antirheumatic agents.
Human peripheral blood mononuclear cells in adherent cultures have been shown to synthesise and secrete collagenase. In the present study we have examined the modulation of collagenase production in these cultures by several antirheumatic agents. Incubation of monocytes in serum free medium with sodium aurothiomalate in concentrations varying from 7.7 X 10(-7) to 7.7 X 10(-3) mol/l resulted in marked dose dependent inhibition of the collagenase production. This inhibition was apparently selective in that total protein synthesis or the viability of the cells were not affected. Similar inhibition of the collagenase production was also noted with auranofin, aurothioglucose, and chloroauric acid. The inhibition with auranofin was achieved with a concentration as low as 7.4 X 10(-8) mol/l. To examine the mechanisms of the inhibition of the collagenase activity induced by sodium aurothiomalate the production of prostaglandin E2 was also measured in the same cell cultures. Sodium aurothiomalate in concentrations greater than 7.7 X 10(-4) mol/l significantly inhibited the prostaglandin E2 production; the prostaglandin E2 production was not inhibited, however, in 7.7 X 10(-5) mol/l concentration, while the collagenase production was reduced by 51.0%. Also, exogenous prostaglandin E2 added to the cultures only slightly reversed the inhibition of the collagenase production by sodium aurothiomalate. Thus the inhibition of collagenase production by sodium aurothiomalate in human adherent mononuclear cell cultures appears to be independent of the inhibition of prostaglandin E2 production. The inhibition of collagenase produced by monocyte-macrophages, as shown here in vitro, may contribute to the clinical efficacy of the compounds tested in the treatment of rheumatoid arthritis.


Relationship between preoperative patency of the inferior mesenteric artery and subsequent occurrence of type II endoleak in patients undergoing endovascular repair of abdominal aortic aneurysms.
The purpose of this study was (1) to find out whether preoperative inferior mesenteric artery (IMA) patency (on radiographic imaging) predicts IMA-related endoleaks after endovascular repair of infrarenal abdominal aortic aneurysms, (2) to determine feasibility of measuring aneurysm sac pressures in patients with endoleaks, and (3) to report early evidence of effective endovascular obliteration of IMA endoleaks. We studied 76 consecutive cases of infrarenal aortic aneurysms that were repaired with an endovascular approach (March 1998-April 1999). There were 13 (17%) endoleaks persistent 30 days after the procedure. Eleven (85%) of these 13 were IMA-related endoleaks, which were documented with selective superior mesenteric artery angiography. The preoperative finding (on computed tomographic scan) of a patent IMA does not always predict an IMA-related endoleak, but results in a statistically and clinically significant higher ratio of patients with IMA-related endoleaks in the immediate postoperative period (24% versus 3%, P <.035). In eight of the 11 patients with persistent IMA-related endoleaks, measurement of intra-aneurysm sac pressures was possible, and six of these patients had systemic pressures within the excluded aneurysm sac. Nine (82%) of 11 IMA-related endoleaks were successfully obliterated by means of selective IMA embolization. Many endoleaks are caused by a patent IMA, and this can result in persistence of systemic pressure within the aneurysm sac. The preoperative finding (on computed tomographic scan) of a patent IMA is a predictor of increased rates of IMA endoleaks, and IMA endoleaks can be successfully obliterated through endovascular procedures, after endovascular abdominal aortic aneurysm repair.


Membrane potential as a modulator of the free intracellular Ca2+ concentration in agonist-activated endothelial cells.
We have used combined patch clamp and fura-2 fluorescence to elucidate the role of membrane potential in the regulation of the cytosolic Ca2+ concentration ([Ca2+]i) in a human umbilical vein derived endothelial cell-line, EA.hy926 (EA cells) stimulated with vasoactive agonists, such as ATP, histamine and bradykinin. This stimulation caused hyperpolarization and sustained Ca2+ plateau in nonclamped cells. Clamping agonist-stimulated cells at negative potentials enhanced the amplitude of this plateau, whereas it was smaller at more depolarized potentials, indicating that Ca2+ influx follows its driving force. Depolarization of the membrane by increasing extracellular K+ or by applying charybdotoxin, a blocker of big conductance Ca2+-dependent K+ channels during agonist stimulation diminished the plateau rise in [Ca2+]i. It is concluded that the membrane potential is an efficient regulator of Ca2+ influx during the plateau phase of agonist-mediated Ca2+ signals. In addition, the modulating effects on Ca2+ signals should be interpreted with caution if the membrane potential of the cells is not controlled.


Production and characterization of monoclonal antibodies to insulin secretory granule membranes.
Monoclonal antibodies to insulin secretory granule membranes were obtained following immunization of mice with granule membranes purified from a rat transplantable insulinoma. The specificities of the antibodies were investigated by using binding assays with different insulinoma subcellular fractions, by indirect immunofluorescence studies with intact and permeabilized cells, and by immunoblotting of granule membrane proteins fractionated by SDS/polyacrylamide-gel electrophoresis. Fifty-six antibodies were characterized initially, and 21 representative cell lines were cloned. The antibodies fell into four categories: (1) binding preferentially to secretory granules, and reacting with a component of approx. 80,000 Da on immunoblots (antigen designated SGM 80); (2) binding preferentially to secretory granules, and reacting with components of approx. 110,000 and 50,000 Da on immunoblots (antigen designated SGM 110); (3) binding preferentially to secretory granules but unreactive on immunoblots; (4) binding to membrane antigen(s) with a widespread intracellular distribution which included granules and plasma membranes. The antigens SGM 80 and SGM 110 were studied in more detail and both were shown to be integral membrane glycoproteins with antigenic determinants located on the internal face of the secretory granule membrane. These antigens were also present in normal rat islets of Langerhans and similar components were detected by immunoblotting in secretory granules from anterior pituitary and adrenal medulla. Proteins which were immunologically related to SGM 80 and SGM 110, but distinct in molecular size, were also identified in liver. It is concluded that secretory granules contain specific components which are restricted in subcellular location but widespread in tissue distribution. The antibodies obtained will be valuable reagents in the further investigation of the biogenesis and turnover of insulin secretory granules.


Application of P450RGS reporter gene system as a bioindicator of sediment PAH contamination in the vicinity of Incheon Harbor, Korea.
Sixty seven sediment samples were collected from Kyeonggi Bay, Korea, including the mouth of Han River, Incheon Harbor, the Namdong industrial complex, and the open sea. Collections were conducted in December, 1995 and samples were maintained frozen (-20 °C) until analysis. Dichloromethane extracts were analysed for their content of CYP1A1-inducing compounds with a P450RGS (reporter gene system) assay, and for polycyclic aromatic hydrocarbons (PAHs). Sediment samples were also analysed for organic carbon (OC) content and grain size, to aid in evaluating the relationship between contamination and physical nature of the sediments. The responses of the P450RGS assay to the sediment extracts were expressed as μg of benzo[a]pyrene toxic equivalents per g dry weight (μg g(-1) BaPTEQ), and these values correlated well (r(2) = 0.624 with total PAHs. BaPTEQ values were also highly correlated with the OC content of the sediments. The determination of P450RGS BaPTEQ is a useful tool, because it is both a rapid and inexpensive means of assessing the potential toxicity of organic compounds in environmental sediment samples. These values represent an estimate of the levels of compounds in the sediment that are potentially available to organisms through chronic exposure to pore water or ingestion of benthic species. We believe BaPTEQ values are more useful than tables of specific PAH concentrations, if the purpose of the investigation is to either obtain a rapid screening of an area or to develop some form of ecological or human health risk assessment.


[Analysis of the Cochrane Review: Cortisteroid Implants for Chronic Non-Infectious Uveitis. Cochrane Database Syst Rev. 2016;2:CD010469].
Non-infectious uveitis is a heterogeneous collection of inflammatory eye diseases and is one of the most important causes of blindness among active adults in developed countries. Inflammation control is crucial in the prevention of ocular structural and functional damage. Regarding acute inflammatory control, corticosteroids are the mainstay of treatment. Several types of intra-ocular corticosteroids have been used with the aim of enhanced efficacy compared to their topical or peri-ocular administration, while minimizing its adverse effects associated with the systemic administration. The purpose of this Cochrane review was to synthetize the available evidence regarding the efficacy and safety of corticosteroid implants in comparison with standard treatment. Due to the heterogeneity in the design of the studies and outcome measures assessed, authors could not conclude that implants are superior to traditional systemic therapy in Non-infectious uveitis. The safety analysis suggested increased risks of post-implant surgery for cataract and high intraocular pressure compared with standard-of-care therapy.


Myonuclear addition to uninjured laryngeal myofibers in adult rabbits.
In normal mature limb skeletal muscle, satellite cells are quiescent and myonuclei do not divide after formation of their associated myofibers in the absence of injury. The possibility of myonuclear addition in uninjured laryngeal myofibers of adult rabbits was investigated in an immunohistochemical pilot study. Bromodeoxyuridine (brdU), a marker for cell division, was administered by intraperitoneal injection over a 12-hour period in rabbits. The number of brdU-positive myonuclei per myofiber was determined on cross sections through the thyroarytenoid (TA) and posterior cricoarytenoid (PCA) muscles. In the TA muscle, 0.13% +/- 0.03% (mean +/- SEM) of the myofibers counted had a brdU-positive nucleus. In the PCA muscle, 0.13% +/- 0.01% of the myofibers counted had a brdU-positive nucleus. Approximately 0.2% and 0.3% of the myofibers of the TA and PCA muscles, respectively, had brdU-positive satellite cells associated with them. Tibialis anterior and pectoralis major muscle controls were negative for brdU-positive myonuclei. These data support the possibility of continuous remodeling in uninjured adult laryngeal myofibers and accentuate the distinct nature of laryngeal muscle relative to limb skeletal muscle in the rabbit model.


Application of meta-transcriptomics and -proteomics to analysis of in situ physiological state.
Analysis of the growth-limiting factor or environmental stressors affecting microbes in situ is of fundamental importance but analytically difficult. Microbes can reduce in situ limiting nutrient concentrations to sub-micromolar levels, and contaminated ecosystems may contain multiple stressors. The patterns of gene or protein expression by microbes in nature can be used to infer growth limitations, because they are regulated in response to environmental conditions. Experimental studies under controlled conditions in the laboratory provide the physiological underpinnings for developing these physiological indicators. Although regulatory networks may differ among specific microbes, there are some broad principles that can be applied, related to limiting nutrient acquisition, resource allocation, and stress responses. As technologies for transcriptomics and proteomics mature, the capacity to apply these approaches to complex microbial communities will accelerate. Global proteomics has the particular advantage that it reflects expressed catalytic activities. Furthermore, the high mass accuracy of some proteomic approaches allows mapping back to specific microbial strains. For example, at the Rifle IFRC field site in Western Colorado, the physiological status of Fe(III)-reducing populations has been tracked over time. Members of a "subsurface clade" within the Geobacter predominated during carbon amendment to the subsurface environment. At the functional level, proteomic identifications produced inferences regarding (i) temporal changes in anabolism and catabolism of acetate, (ii) the onset of N(2) fixation when N became limiting, and (iii) expression of phosphate transporters during periods of intense growth. The application of these approaches in situ can lead to discovery of novel physiological adaptations.


[Tyr1]-nociceptin, a novel nociceptin analog, decreases systemic arterial pressure by a naloxone-insensitive mechanism in the rat.
The heptadecapeptide nociceptin, also known as Orphanin FQ, is a newly discovered endogenous ligand for the opioid-like G-protein-coupled receptor ORL1. In the present study, responses to a novel nociceptin analog, [Tyr1]-nociceptin, were investigated in the systemic vascular bed of the rat. [Tyr1]-nociceptin induced dose-related decrease in systemic arterial pressure when injected in doses of 1-30 nmol/kg i.v. In terms of relative vasodepressor activity, [Tyr1]-nociceptin was comparable in potency to nociceptin. The decreases in systemic arterial pressure in response to [Tyr1]-nociceptin were not altered by the opioid receptor antagonist naloxone in a dose that reduced responses to Met-enkephalin. The results of the present study show that vasodepressor responses to [Tyr1]-nociceptin are not mediated by the activation of a naloxone-sensitive opioid receptor and are not dependent on the presence of Phe at the N-terminus of the nociceptin sequence.


Molecular mechanism for the interaction between gibberellin and brassinosteroid signaling pathways in Arabidopsis.
Plant development is modulated by the convergence of multiple environmental and endogenous signals, and the mechanisms that allow the integration of different signaling pathways is currently being unveiled. A paradigmatic case is the concurrence of brassinosteroid (BR) and gibberellin (GA) signaling in the control of cell expansion during photomorphogenesis, which is supported by physiological observations in several plants but for which no molecular mechanism has been proposed. In this work, we show that the integration of these two signaling pathways occurs through the physical interaction between the DELLA protein GAI, which is a major negative regulator of the GA pathway, and BRASSINAZOLE RESISTANT1 (BZR1), a transcription factor that broadly regulates gene expression in response to BRs. We provide biochemical evidence, both in vitro and in vivo, indicating that GAI inactivates the transcriptional regulatory activity of BZR1 upon their interaction by inhibiting the ability of BZR1 to bind to target promoters. The physiological relevance of this interaction was confirmed by the observation that the dominant gai-1 allele interferes with BR-regulated gene expression, whereas the bzr1-1D allele displays enhanced resistance to DELLA accumulation during hypocotyl elongation. Because DELLA proteins mediate the response to multiple environmental signals, our results provide an initial molecular framework for the integration with BRs of additional pathways that control plant development.


Genome-Based Metabolic Reconstruction of a Novel Uncultivated Freshwater Magnetotactic coccus "Ca. Magnetaquicoccus inordinatus" UR-1, and Proposal of a Candidate Family "Ca. Magnetaquicoccaceae".
Magnetotactic bacteria are widely represented microorganisms that have the ability to synthesize magnetosomes. The magnetotactic cocci of the order Magnetococcales are the most frequently identified, but their classification remains unclear due to the low number of cultivated representatives. This paper reports the analysis of an uncultivated magnetotactic coccus UR-1 collected from the Uda River (in eastern Siberia). Genome analyses of this bacterium and comparison to the available Magnetococcales genomes identified a novel species called "Ca. Magnetaquicoccus inordinatus," and a delineated candidate family "Ca. Magnetaquicoccaceae" within the order Magnetococcales is proposed. We used average amino acid identity values <55-56% and <64-65% as thresholds for the separation of families and genera, respectively, within the order Magnetococcales. Analyses of the genome sequence of UR-1 revealed a potential ability for a chemolithoautotrophic lifestyle, with the oxidation of a reduced sulfur compound and carbon assimilation by rTCA. A nearly complete magnetosome genome island, containing a set of mam and mms genes, was also identified. Further comparative analyses of the magnetosome genes showed vertical inheritance as well as horizontal gene transfer as the evolutionary drivers of magnetosome biomineralization genes in strains of the order Magnetococcales.


Soft tissue sarcomas of the foot and ankle: impact of unplanned excision, limb salvage, and multimodality therapy.
Foot and ankle sarcomas have historically been treated with amputation because of the difficulty in achieving local disease control and maintaining a functional foot. Potential opportunities for limb salvage may be further compromised by unplanned excisions. We reviewed 52 consecutive patients with soft tissue sarcomas of the foot and ankle and analyzed the impact of planned versus unplanned initial excision, limb salvage, and multimodality therapy on treatment and outcomes. Unplanned excisions had been performed in 29 (55.8%) patients. Limb salvage was performed in 38 patients, with 14 requiring free soft tissue transfers. At an average followup of 99 (range, 24 to 216) months, the 5-year overall survival estimate was 76.3%. Although not statistically significant, we noted clinically relevant potential differences in local recurrence-free, disease-free, and oncologic survival between the planned and unplanned excision groups. Seven patients (13.5%) had a local recurrence, five of these following an unplanned excision. Functional scores averaged 83.2% for all patients which were similar between planned versus unplanned and amputation versus limb salvage groups. Significantly more patients with unplanned excisions required free flaps for limb salvage (p = 0.017) and received adjuvant radiotherapy (p = 0.0004). Unplanned surgery for soft tissue sarcomas of the foot and ankle often results in the need for more aggressive surgery and/or adjuvant radiotherapy and may impact oncologic outcomes, but does not necessarily portend worse functional outcomes. Multimodal therapy and judicious use of soft tissue flap reconstruction allows limb salvage in most patients with favorable outcomes.


Convergence of CONSTITUTIVE PHOTOMORPHOGENESIS 1 and PHYTOCHROME INTERACTING FACTOR signalling during shade avoidance.
Shade-avoidance responses require CONSTITUTIVE PHOTOMORPHOGENESIS 1 (COP1) but the mechanisms of action of COP1 under shade have not been elucidated. Using simulated shade and control conditions, we analysed: the transcriptome and the auxin levels of cop1 and phytochrome interacting factor 1 (pif1) pif3 pif4 pif5 (pifq) mutants; the dynamics of ELONGATED HYPOCOTYL 5 (HY5) and LONG HYPOCOTYL IN FAR-RED (HFR1) proteins; and the epistatic relationships between cop1 and pif3, pif4, pif5, hy5 and hfr1 mutations in Arabidopsis thaliana. Despite severely impaired shade-avoidance responses, only a few genes that responded to shade in the wild-type failed to do so in cop1. Shade enhanced the convergence between cop1 and pifq transcriptomes, mainly on shade-avoidance marker genes. Shade failed to increase auxin levels in cop1. Residual shade avoidance in cop1 was not further reduced by the pif3, pif4 or pif5 mutations, suggesting convergent pathways. HFR1 stability decreased under shade in a COP1-dependent manner but shade increased HY5 stability. The cop1 mutant retains responses to shade and is more specifically impaired in shade avoidance. COP1 promotes the degradation of HFR1 under shade, thus increasing the ability of PIFs to control gene expression, increase auxin levels and promote stem growth.


A 3-year Health Care Coalition Experience in Advancing Hospital Evacuation Preparedness.
This report outlines a 3-year health care coalition effort to advance and test community capacity for a large-scale hospital evacuation. The multi-year effort utilized a variety workshops, seminars, webinars, tabletops, functional exercises, and culminated with a full-scale exercise testing hospital evacuation. While most hospital evacuation exercises focus on internal movement of patients, this exercise process tested command-level decision making and it tested external partners such as transportation agencies, law enforcement, receiving hospitals, and local emergency management. This process delivered key coalition-building activities and offered a variety of training and exercise opportunities to assist a number of organizations, all at different stages of hospital evacuation planning. The 2012 Hospital Preparedness Program outlined the incorporation of health care coalition activities to transform individual organization preparedness to community-level readiness. This report outlines a health care coalition effort to deliver training and exercises to advance community capacity for a large-scale hospital evacuation. Lowe JJ , Hansen KF , Sanger KK , Obaid JM . A 3-year health care coalition experience in advancing hospital evacuation preparedness. Prehosp Disaster Med. 2016;31(6):658-662.


Serum concentrations of glial fibrillary acidic protein (GFAP) do not indicate tumor recurrence in patients with glioblastoma.
Recent studies identified serum concentrations of the astroglial protein glial fibrillary acidic protein (GFAP) to be indicative of glioblastoma (GBM) in patients with newly diagnosed space occupying cerebral mass lesions. Until now, no data is available whether GFAP serum concentrations decrease after first therapy and whether GFAP may be used as a predictor of survival and an indicator of tumor recurrence. In this prospective study, we included 44 patients with a single space occupying cerebral mass lesion suspicious for GBM. GBM was histopathologically proven in 33 cases. After initial therapy, patients were followed up until tumor recurrence (defined according to the RANO criteria) or death (maximum observation period 78 weeks). Blood was sampled on a regular basis, and GFAP serum levels were determined using an immunofluorescence assay. Prior to any intervention, 14 of the 33 GBM patients had elevated GFAP serum concentrations (median 0.25 µg/L, interquartile range 0.13-0.53), whereas only one out of 11 patients having other tumor entities revealed a slightly increased GFAP serum level (0.06 µg/L). Following surgery (i.e., biopsy, full or partial resection), all initially GFAP positive GBM patients showed decreased serum concentrations. During the follow-up period, we found a minimal GFAP increase in one patient only (0.04 µg/L; week 52), although 23 out of 31 available GBM patients developed tumor progression or died. No difference was found regarding the survival rate and the time to tumor recurrence between initially GFAP positive and GFAP negative GBM patients. In GBM patients, initially elevated GFAP serum concentrations decrease after the first diagnostic or therapeutic intervention. GFAP was not predictive for tumor recurrence.


Topiramate effects on the EEG and alertness in healthy volunteers: a different profile of antiepileptic drug neurotoxicity.
Previous quantitative EEG (QEEG) studies of carbamazepine (CBZ), oxcarbazepine (OXC), and phenytoin (PHT) revealed a pattern of EEG slowing and an increase in drowsiness on the awake maintenance task (AMT). EEG slowing has been shown to correlate with negative effects on cognitive tests. Topiramate (TPM) is a novel AED with relatively large negative effects on cognitive function. We tested the hypothesis that TPM would induce significant slowing of EEG background rhythms and an increase in AMT drowsiness. Forty healthy volunteers were randomized to TPM, gabapentin (GBP), or placebo. Doses were escalated as tolerated to a maximum of 400mg/day for TPM or 3600 mg/day for GBP, over a 10-week period, followed by a minimum 2-week plateau period. Volunteers underwent an EEG, cognitive tests, and the AMT prior to starting an AED and again 12 weeks later. The EEG was captured using a structured recording protocol and quantified using the fast Fourier transform. Four target measures were derived from the averaged occipital electrodes (peak frequency of the dominant posterior rhythm, median frequency, percentage theta, and percentage delta). Test-retest changes for all measures were scored against similar test-retest distributions previously obtained from untreated healthy volunteers. TPM produced no significant change in any of the four target EEG measures or on the AMT, even though several target cognitive tests revealed moderate or greater negative effects. There were also no significant changes in the placebo group. GBP slowed the peak and median frequency EEG measures and increased the percentage of theta and delta activity. Neither TPM, GBP, nor placebo caused a significant increase in drowsiness on the AMT. TPM has a unique neurotoxicity profile. It has no effect on EEG background measures or on the AMT, but induces moderate to large negative changes in many cognitive test scores. This profile differs from those of CBZ, OXC, PHT, and GBP.


A comparison of carbohydrate metabolism after sucrose, sorbitol, and fructose meals in normal and diabetic subjects.
Sucrose, sorbitol, and fructose (35 g) were fed to normal and diabetic subjects as a component of a 400-calorie breakfast. In both normal and diabetic subjects, the mean peak increment in plasma glucose was highest after the sucrose meals (44.0 mg/dl for normal subjects; 78.0 mg/dl for diabetic subjects); lowest after sorbitol meals (9.3 mg/dl for normal subjects; 32.3 mg/dl for diabetic subjects); and intermediate after the fructose meals (29.0 mg/dl for normal subjects; 48.0 mg/dl for diabetic subjects). In normal subjects, the mean peak increment of plasma immunoreactive insulin followed a similar pattern, but in diabetic subjects there was no significant difference between the three groups. We conclude that fructose or sorbitol, given as part of a meal, results in lower glucose levels in both normal and diabetic subjects, but that the latter is not related to a difference in insulin release.


Disease2Vec: Encoding Alzheimer's Progression via Disease Embedding Tree.
For decades, a variety of predictive approaches have been proposed and evaluated in terms of their prediction capability for Alzheimer's Disease (AD) and its precursor - mild cognitive impairment (MCI). Most of them focused on prediction or identification of statistical differences among different clinical groups or phases, especially in the context of binary or multi-class classification. The continuous nature of AD development and transition states between successive AD related stages have been typically overlooked. Though a few progression models of AD have been studied recently, they were mainly designed to determine and compare the order of specific biomarkers. How to effectively predict the individual patient's status within a wide spectrum of continuous AD progression has been largely understudied. In this work, we developed a novel learning-based embedding framework to encode the intrinsic relations among AD related clinical stages by a set of meaningful embedding vectors in the latent space (Disease2Vec). We named this process as disease embedding. By Disease2Vec, our framework generates a disease embedding tree (DETree) which effectively represents different clinical stages as a tree trajectory reflecting AD progression and thus can be used to predict clinical status by projecting individuals onto this continuous trajectory. Through this model, DETree can not only perform efficient and accurate prediction for patients at any stages of AD development (across five fine-grained clinical groups instead of typical two groups), but also provide richer status information by examining the projecting locations within a wide and continuous AD progression process. (Code will be available: https://github.com/qidianzl/Disease2Vec.).


1p34.3 deletion involving GRIK3: Further clinical implication of GRIK family glutamate receptors in the pathogenesis of developmental delay.
A growing body of evidence suggests an association between microdeletion/microduplication and schizophrenia/intellectual disability. Abnormal neurogenesis and neurotransmission have been implicated in the pathogenesis of these neuropsychiatric and neurodevelopmental disorders. The kainate/AMPA-type ionotropic glutamate receptor (GRIK = glutamate receptor, ionotropic, kainate) plays a critical role in synaptic potentiation, which is an essential process for learning and memory. Among the five known GRIK family members, haploinsufficiency of GRIK1, GRIK2, and GRIK4 are known to cause developmental delay, whereas the roles of GRIK3 and GRIK5 remain unknown. Herein, we report on a girl who presented with a severe developmental delay predominantly affecting her language and fine motor skills. She had a 2.6-Mb microdeletion in 1p34.3 involving GRIK3, which encodes a principal subunit of the kainate-type ionotropic glutamate receptor. Given its strong expression pattern in the central nervous system and the biological function of GRIK3 in presynaptic neurotransmission, the haploinsufficiency of GRIK3 is likely to be responsible for the severe developmental delay in the proposita. A review of genetic alterations and the phenotypic effects of all the GRIK family members support this hypothesis. The current observation of a microdeletion involving GRIK3, a kainate-type ionotropic glutamate receptor subunit, and the neurodevelopmental manifestation in the absence of major dysmorphism provides further clinical implication of the possible role of GRIK family glutamate receptors in the pathogenesis of developmental delay.


Early adolescents' motivations to defend victims in school bullying and their perceptions of student-teacher relationships: A self-determination theory approach.
The aim of this study was to investigate whether various dimensions of student-teacher relationships were associated with different types of motivation to defend victims in bullying and to determine the association between these types of motivations and various bystander behaviors in bullying situations among early adolescents in Italy. Data were collected from 405 Italian adolescents who completed a survey in their classroom. Results showed that warm student-teacher relationships were positively associated with defending victims and with autonomous motivation to defend victims. In contrast, conflictual student-teacher relationships were positively associated with passive bystanding and with extrinsic motivation to defend victims. Different forms of motivation to defend were found to be mediators between student-teacher relationship qualities and bystander behaviors in school bullying. Our findings suggest that teachers should build warm and caring student-teacher relationships to enhance students' autonomous motivation to defend victims of bullying as well as their inclination to defend the victims in practice.


Survey of susceptibilities of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis isolates to 26 antimicrobial agents: a prospective U.S. study.
An antimicrobial susceptibility surveillance study of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis isolates was performed during the winter of 1996-1997 in order to determine their susceptibilities to 5 fluoroquinolones and 21 other antimicrobial agents. Broth microdilution MICs were determined for 2, 752 isolates from 51 U.S. medical centers. Of the 1,276 S. pneumoniae isolates, 64% were susceptible, 17% were intermediate, and 19% were highly resistant to penicillin. On the basis of the MICs at which 90% of isolates are inhibited and modal MICs, the hierarchy of the five fluoroquinolones from most to least active was grepafloxacin > sparfloxacin > levofloxacin = ciprofloxacin > ofloxacin. For S. pneumoniae isolates for which penicillin MICs were elevated, the MICs of the cephalosporins, macrolides, clindamycin, tetracycline, and trimethoprim-sulfamethoxazole were also elevated, but the MICs of the fluoroquinolones, vancomycin, and rifampin were not. The prevalence of penicillin-susceptible pneumococci varied by U.S. Bureau of the Census region (range, 44% in the East South Central region to 75% in the Pacific region). In addition, S. pneumoniae isolates from blood were significantly more susceptible to penicillin than those from respiratory, ear, or eye specimens, and pneumococci from patients </=2 years old were significantly more resistant to penicillin than those from older patients (by chi-square analysis, P < 0.05). beta-Lactamase was produced by 35% of H. influenzae isolates and 93% of M. catarrhalis isolates, resulting in increased MICs of amoxicillin and certain cephalosporins. We noted that the antimicrobial resistance patterns of S. pneumoniae isolates, which correlate with the penicillin susceptibility phenotype, vary by site of infection, age group of the patient, and geographic source of the isolate.


RESEARCH: Managing Mountainous Degraded Landscapes After Farmland Abandonment in the Central Spanish Pyrenees
/ Plant succession and pasture resources have been studied in abandoned fields of the central Spanish Pyrenees, in an environment severely affected by strong demographic pressure in the past. Several hydromorphological features (runoff and sediment yield) were also analyzed for different environments of the abandoned fields, in order to forecast the effects of their reclamation and transformation into areas for livestock use. The availability and accessibility of pastures as well as soil and water conservation is related to the process of colonization of Genista scorpius. Under a dense shrub cover both runoff and sediment yield are strongly controlled. As the shrub cover becomes open, sediment yield and runoff increase greatly. A dense herbaceous cover yields high runoff coefficients but moderate soil losses. From the results obtained, the possibility of abandoned field reclamation by means of selective clearing of scrub is discussed.KEY WORDS: Abandoned fields; Plant succession; Degraded environments; Soil erosion; Runoff; Spanish Pyrenees


Identification of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) as a novel target of bisphenol A.
Bisphenol A (BPA) forms the backbone of plastics and epoxy resins used to produce packaging for various foods and beverages. BPA is also an estrogenic disruptor, interacting with human estrogen receptors (ER) and other related nuclear receptors. Nevertheless, the effects of BPA on human health remain unclear. The present study identified DNA-dependent protein kinase catalytic subunit (DNA-PKcs) as a novel BPA-binding protein. DNA-PKcs, in association with the Ku heterodimer (Ku70/80), is a critical enzyme involved in the repair of DNA double-strand breaks. Low levels of DNA-PK activity are previously reported to be associated with an increased risk of certain types of cancer. Although the Kd for the interaction between BPA and a drug-binding mutant of DNA-PKcs was comparatively low (137 nM), high doses of BPA were required before cellular effects were observed (100-300 μM). The results of an in vitro kinase assay showed that BPA inhibited DNA-PK kinase activity in a concentration-dependent manner. In M059K cells, BPA inhibited the phosphorylation of DNA-PKcs at Ser2056 and H2AX at Ser139 in response to ionizing radiation (IR)-irradiation. BPA also disrupted DNA-PKcs binding to Ku70/80 and increased the radiosensitivity of M059K cells, but not M059J cells (which are DNA-PKcs-deficient). Taken together, these results provide new evidence of the effects of BPA on DNA repair in mammalian cells, which are mediated via inhibition of DNA-PK activity. This study may warrant the consideration of the possible carcinogenic effects of high doses of BPA, which are mediated through its action on DNA-PK.


Counteracting Immunosenescence-Which Therapeutic Strategies Are Promising?
Aging attenuates the overall responsiveness of the immune system to eradicate pathogens. The increased production of pro-inflammatory cytokines by innate immune cells under basal conditions, termed inflammaging, contributes to impaired innate immune responsiveness towards pathogen-mediated stimulation and limits antigen-presenting activity. Adaptive immune responses are attenuated as well due to lowered numbers of naïve lymphocytes and their impaired responsiveness towards antigen-specific stimulation. Additionally, the numbers of immunoregulatory cell types, comprising regulatory T cells and myeloid-derived suppressor cells, that inhibit the activity of innate and adaptive immune cells are elevated. This review aims to summarize our knowledge on the cellular and molecular causes of immunosenescence while also taking into account senescence effects that constitute immune evasion mechanisms in the case of chronic viral infections and cancer. For tumor therapy numerous nanoformulated drugs have been developed to overcome poor solubility of compounds and to enable cell-directed delivery in order to restore immune functions, e.g., by addressing dysregulated signaling pathways. Further, nanovaccines which efficiently address antigen-presenting cells to mount sustained anti-tumor immune responses have been clinically evaluated. Further, senolytics that selectively deplete senescent cells are being tested in a number of clinical trials. Here we discuss the potential use of such drugs to improve anti-aging therapy.


Mitochondrial pathway is involved in hydrogen-peroxide-induced apoptotic cell death of oligodendrocytes.
Oligodendrocytes, the myelin-forming cells of the CNS, are specifically sensitive to oxidative stress and respond by the onset of programmed cell death (PCD). To further unravel the molecular events underlying their enhanced susceptibility, we have investigated whether mitochondrial damage occurs during oxidative stress-induced PCD in cultured rat brain oligodendrocytes. Mitochondria are considered as a central control point of apoptosis, and mitochondrial dysfunction has been linked to neurodegenerative disease. Upon a number of stimuli through the release of cytochrome c, they coordinate caspase activation, causing morphological and biochemical changes associated with PCD. Oxidative stress was exerted by the application of hydrogen peroxide. The data show that hydrogen peroxide-induced apoptosis in oligodendrocytes involves mitochondrial damage and cytochrome c release and is accompanied by the activation of the death-related caspases 3 and 9. Concomitantly, the activation and nuclear translocation of extracellular signal regulated kinases ERK1,2 are observed, which have been implicated to participate in the regulation of cell death and survival. DNA fragmentation could not be attenuated by the ERK1,2 inhibitor PD 98059, indicating that the ERK1,2- pathway in oligodendrocytes may be involved in the initial survival response after exposure to stressful stimuli.


Exercise Predicts Long-Term Weight Loss in Women With Class 1 and Class 2 Obesity Through Effects on Emotional Eating and its Correlates.
The relationship between exercise and long-term weight loss is definitive. However, in deconditioned individuals, the basis of that relationship beyond minimal energy expenditures is unclear. Effects emanating from exercise's proposed association with changes in emotional eating and its psychosocial correlates were tested. Women with class 1 (n = 67) and class 2 (n = 53) obesity, participating in a new community-based weight-loss treatment based on tenets of social cognitive theory, were assessed at baseline and Months 3, 6, 12, and 24 on weight and measures of exercise outputs, emotional eating, eating self-regulation, and negative mood. An exercise-support component was provided for 2 months prior to group nutrition-change sessions. Improvements from baseline were significant on all measures but did not significantly differ by group. Although not for the initial 3 months, changes in exercise significantly predicted weight changes over 6, 12, and 24 months from baseline. However, only 26%-31% of the lost weight was directly attributable to exercise-related energy expenditures. Changes in emotional eating significantly mediated the exercise-weight-loss relationship, and changes in self-regulation and mood significantly mediated the exercise-emotional eating relationship. Findings supported exercise's role in long-term weight loss primarily through psychosocial factors.


Progressive degradation and subsequent refinement of acoustic representations in the adult auditory cortex.
Correlated neuronal activity is believed to play an important role in refining and maintaining cortical circuitry during early development. Here we provide evidence that globally and locally correlated activity mediate different forms of adult plasticity. Pulses of broad-spectrum noise were used to activate time-locked responses across large areas of the rat auditory cortex, globally synchronizing cortical activity. Brief tone pips were used to activate relatively small groups of neurons, generating locally correlated activity. Pairing pulsed noises with nucleus basalis (NB) stimulation in awake rats for 4 weeks broadened spectral tuning, disrupted tonotopic maps, and reduced spontaneous discharge correlation in the primary auditory cortex (AI), as examined under anesthesia. Those effects caused AI neurons to appear qualitatively similar to neurons in nonprimary auditory fields of naive animals. Subsequent pairing of tone pips with NB stimulation for a period of 4 weeks completely reversed these effects induced by previous noise-NB pairing. These findings further demonstrate that the adult auditory cortex retains a substantial capacity for receptive field plasticity and tonotopic map reorganization and that locally correlated activity plays an important role in plasticity in the adult, as in the developing cortex.


Alveolar soft-part sarcoma: Primary metastatic disease and metastatic relapse occurring during long-term follow-up: Treatment results of four Cooperative Weichteilsarkom Studiengruppe (CWS) trials and one registry.
Patients with metastatic alveolar soft-part sarcoma (ASPS) are known to have a very poor prognosis. Little is known about best treatment of primary metastatic disease (MD) or relapsed metastatic disease (rMD). Patients with localized disease (LD), primary MD, and metastatic recurrence after complete remission (CR) treated within the CWS-86, -91, -96, -2002P trials and the recent registry SoTiSaR (1985-2016) were analyzed. Fifteen of 61 patients had primary metastases at initial diagnosis at the age of 14.6 years (range, 7.8-19.7). Nine of 46 patients with initial LD suffered of rMD at a median age of 9.9 years (range, 3.5-30), 3.75 years (0.75-21) after CR of primary disease. Complete resection (microscopically or macroscopically) was possible in 2 of 15 patients with MD and in 5 of 9 with rMD. RT was administered in 4 of 15 MD and 1 of 9 rMD. Chemotherapy was administered to 11 of 15 MD and 3 of 9 rMD, targeted therapy to 3 of 15 MD and 1 of 9 rMD. Median time to progression of patients treated with targeted therapy (n = 4), CHT (n = 14), and resection only (n = 6) was 56, 17, and 23 months, respectively. The 5-year event-free survival and overall survival (OS) rates were 19.8% and 61%, respectively, for patients with MD compared with 79% and 98% for patients with LD. The 5-year progression-free survival and OS were 67% and 100% for patients with rMD. Complete tumor resection correlates with long-term survival in patients with primary and relapsed MD.


Study of the adenovascular body of the posterior part of the nasal septum.
The mucosa of the posterior part of the nasal septum is studied by in vivo observations in 40 seemingly healthy adults, vascular injection of two anatomical specimens rendered transparent, histological studies of serial sections of five anatomical specimens, and macroscopic and histological examination of specimens obtained from 20 cadavers. The presence of an "adenovascular" body was noted in the posterior part of the nasal septum; this body occupies about one fifth of the total area of this region. This study shows that this is a normal structure of the nose and suggests that its name be changed to adenovascular body of the posterior part of the nasal septum.


Zygosity diagnosis in young twins by questionnaire for twins' mothers and twins' self-reports.
The present study deals with the determination of zygosity in twins of childhood age by simple questionnaire. The subjects were 224 twin pairs and their mothers, consisting of 159 monozygotic and 65 same-sex dizygotic pairs, identified by genetic markers including DNA samples. Mothers of twins responded to 19 questionnaire items dealing with twin similarity in 16 items about physical features and 3 items about the degree of similarity and frequency of being mistaken (confusion of identity) when twins were about 1 year of age. The twins themselves responded to three questionnaire items dealing with only confusion of identity items. The results of stepwise logistic regression analysis were as follows: the total accuracy of the mothers' questionnaire was 91.5% when using only the items dealing with confusion of identity. This accuracy was slightly lower than that obtained by twins' self-reports dealing with nearly the same question items of confusion of identity, answered by both twins separately with 93.3% accuracy. The total accuracy of mothers' questionnaire responses rose to 95.1% when we used all 19 items. In addition to "the frequency of being mistaken", two physical features, namely "shape of fingers" and "shape of eyebrow", were very informative. In conclusion, twin zygosity can be estimated by the use of the mothers' simple questionnaire with sufficient accuracy even in very young twins about 1 year of age.


Thermodiffusion of citrate-coated γ-Fe2O3 nanoparticles in aqueous dispersions with tuned counter-ions - anisotropy of the Soret coefficient under a magnetic field.
Under a temperature gradient, the direction of thermodiffusion of charged γ-Fe2O3 nanoparticles (NPs) depends on the nature of the counter-ions present in the dispersion, resulting in either a positive or negative Soret coefficient. Various counter-ions are probed in finely tuned and well characterized dispersions of citrate-coated NPs at comparable concentrations of free ionic species. The Soret coefficient ST is measured in stationary conditions together with the mass-diffusion coefficient Dm using a forced Rayleigh scattering method. The strong interparticle repulsion, determined by SAXS, is also attested by the increase of Dm with NP volume fraction Φ. The Φ-dependence of ST is analyzed in terms of thermophoretic and thermoelectric contributions of the various ionic species. The obtained single-particle thermophoretic contribution of the NPs (the Eastman entropy of transfer ŝNP) varies linearly with the entropy of transfer of the counter-ions. This is understood in terms of electrostatic contribution and of hydration of the ionic shell surrounding the NPs. Two aqueous dispersions, respectively, with ST > 0 and with ST < 0 are then probed under an applied field H[combining right harpoon above], and an anisotropy of Dm and of ST is induced while the in-field system remains monophasic. Whatever the H[combining right harpoon above]-direction (parallel or perpendicular to the gradients and ), the Soret coefficient is modulated keeping the same sign as in zero applied field. In-field experimental determinations are well described using a mean field model of the interparticle magnetic interaction.


Investigating Potential Dose-Response Relationships between Vitamin D Status and Cognitive Performance: A Cross-Sectional Analysis in Middle- to Older-Aged Adults in the Busselton Healthy Ageing Study.
Low vitamin D status has been linked to adverse cognitive outcomes in older adults. However, relationships at higher levels remain uncertain. We aimed to clarify patterns of association between vitamin D status and cognitive performance, using flexible regression methods, in 4872 middle- to older-aged adults (2678 females) from the Busselton Healthy Ageing Study. Cross-sectional associations of serum levels of 25-hydroxyvitamin D (25OHD) and performance in cognitive domains were modelled using linear regression and restricted cubic splines, controlling for demographic, lifestyle, and health factors. Mean ± SD serum 25OHD levels were 78 ± 24 nM/L for women and 85 ± 25 nM/L for men. Increasing levels in women were associated with better global cognition (linear trend, p = 0.023) and attention accuracy (continuity of attention), with improvement in the latter plateauing around levels of 80 nM/L (nonlinear trend, p = 0.035). In men, increasing levels of serum 25OHD were associated with better attention accuracy (linear trend, p = 0.022), but poorer semantic verbal fluency (linear trend, p = 0.025) and global cognition (nonlinear trend, p = 0.015). We identified patterns of association between serum 25OHD levels and cognitive performance that may reflect early dose-response relationships, particularly in women. Longitudinal analyses extending through to older ages may help to clarify the nature, strength, and temporality of these relationships.


The principle of stationary action in biophysics: stability in protein folding.
We conceptualize protein folding as motion in a large dimensional dihedral angle space. We use Lagrangian mechanics and introduce an unspecified Lagrangian to study the motion. The fact that we have reliable folding leads us to conjecture the totality of paths forms caustics that can be recognized by the vanishing of the second variation of the action. There are two types of folding processes: stable against modest perturbations and unstable. We also conjecture that natural selection has picked out stable folds. More importantly, the presence of caustics leads naturally to the application of ideas from catastrophe theory and allows us to consider the question of stability for the folding process from that perspective. Powerful stability theorems from mathematics are then applicable to impose more order on the totality of motions. This leads to an immediate explanation for both the insensitivity of folding to solution perturbations and the fact that folding occurs using very little free energy. The theory of folding, based on the above conjectures, can also be used to explain the behavior of energy landscapes, the speed of folding similar to transition state theory, and the fact that random proteins do not fold.


[Realization of Long Bone Stitching Technology in DR Chest Radiography System].
Using a common DR chest radiography system to realize a long bone stitching technology. Introduce the role of long bone stitching technology in medical diagnosis and treatment, and the principle of long bone stitching technology to make a long bone stitching radiographic device, and combine with the chest radiography system to take the long bone stitching image experiment. The hospitals of class Ⅱ (or more lower levels) can realize the long bone stitching technology using a common DR chest radiography system. Using this technology can save the hospital costs, reduce the burden on patients, achieve good social and economic benefits.


Removing structural disorder from oriented TiO2 nanotube arrays: reducing the dimensionality of transport and recombination in dye-sensitized solar cells.
We report on the influence of morphological disorder, arising from bundling of nanotubes (NTs) and microcracks in films of oriented TiO2 NT arrays, on charge transport and recombination in dye-sensitized solar cells (DSSCs). Capillary stress created during evaporation of liquids from the mesopores of dense TiO2 NT arrays was of sufficient magnitude to induce bundling and microcrack formation. The average lateral deflection of the NTs in the bundles increased with the surface tension of the liquids and with the film thicknesses. The supercritical CO2 drying technique was used to produce bundle-free and crack-free NT films. Charge transport and recombination properties of sensitized films were studied by frequency-resolved modulated photocurrent/photovoltage spectroscopies. Transport became significantly faster with decreased clustering of the NTs, indicating that bundling creates additional pathways via intertube contacts. Removing such contacts alters the transport mechanism from a combination of one and three dimensions to the expected one dimension and shortens the electron-transport pathway. Reducing intertube contacts also resulted in a lower density of surface recombination centers by minimizing distortion-induced surface defects in bundled NTs. A causal connection between transport and recombination is observed. The dye coverage was greater in the more aligned NT arrays, suggesting that reducing intertube contacts increases the internal surface area of the films accessible to dye molecules. The solar conversion efficiency and photocurrent density were highest for DSSCs incorporating films with more aligned NT arrays owing to an enhanced light-harvesting efficiency. Removing structural disorder from other materials and devices consisting of nominally one-dimensional architectures (e.g., nanowire arrays) should produce similar effects.


The periovulatory period in cattle: progesterone, prostaglandins, oxytocin and ADAMTS proteases.
Ovulation has long been recognized as one of the most dramatic reproductive processes. Decades of research on how the LH/FSH surge leads to ovulation have made it clear that the surge induces a very complex cascade of changes. Studies of genetically modified mice have pointed to progesterone (P4) and its receptor (PGR) and the prostaglandins (PGs) as critical components of the ovulatory cascade. In cattle, the gonadotropin surge also induces oxytocin (OT), which does not appear to increase in rodent periovulatory follicles. This review is an attempt to summarize studies by our laboratory on the temporal patterns, roles, regulation, and interrelationships among P4/PGR, PGs, and OT in bovine periovulatory follicles. Most of these results are based on an experimental model in which the dominant follicle of the first follicular wave of the estrous cycle is induced to develop into a preovulatory follicle by injection of PGF(2α) on Day 6 of the cycle, followed 36 h later by an injection of GnRH to induce the LH/FSH surge. The results suggest that the effects of the gonadotropin surge on PG production by bovine granulosa cells are mediated by the gonadotropin-induced increase in intrafollicular P4 and that P4 acts by binding to its nuclear receptor and increasing the abundance of mRNA for the enzyme PTGS2 (COX-2). Our data thus far also support the hypothesis that PGs, especially PGE(2), can stimulate progesterone secretion by both follicular cell types and suggest a positive feedback relationship between P4/PGR and the PGs. Additional results suggest a positive feedback loop between P4/PGR and OT. The finding that levels of mRNA for several ADAMTS proteases are regulated by the LH/FSH surge in vivo and by P4/PGR and/or PGs in vitro suggests a role for this family of proteases in remodeling the bovine ovulatory follicle in preparation for ovulation and the formation of the corpus luteum. It is important to remember that a process essential for reproduction, such as ovulation, may involve redundant mechanisms and that these mechanisms may have evolved differently from rodents in larger mammalian species, such as ruminants and humans.


Development of a bio-inspired mechatronic chest wall simulator for evaluating the performances of opto-electronic plethysmography.
Instrumented gait analysis based on optoelectronic systems is an expensive technique used to objectively measure the human movement features and it is generally considered as the gold standard. Opto-electronic plethysmography (OEP) is a particular motion analysis system able to: (i) determine chest wall kinematic via the evaluation of marker displacements placed on the thorax and (ii) compute respiratory volumes during breathing. The aim of this work is to describe the performances of a custom made, bio-inspired, mechatronic chest wall simulator (CWS), specifically designed to assess the metrological performances of the OEP system. The design of the simulator is based on the chest wall kinematic analysis of three healthy subjects previously determined. Two sets of experiments were carried out: (i) to investigate the CWS dynamic response using different target displacements (1 - 12 mm), and (ii) to assess the CWS accuracy and precision in simulating quite breathing, covering the physiological range of respiratory frequency and tidal volume. Results show that the CWS allows simulating respiratory frequency up to ~ 60 bpm. The difference between the actual displacement and the set one is always < 9 μm. The precision error, expressed as the ratio between measurement uncertainty and the actual displacement, is lower than 0.32 %. The observed good performances permit to consider the CWS prototype feasible to be employed for assessing the performances of OEP system in periodical validation routines.


Characterization of vitreous B-cell infiltrates in patients with primary ocular lymphoma, using CDR3 size polymorphism analysis of antibody transcripts.
Histopathology usually cannot be performed and cytology is unfortunately frequently insufficient to confirm a suspicion of primary intraocular lymphoma (PIOL). The purpose of this study was to evaluate the Immunoscope technique for the identification of ocular B-cell monoclonal infiltrates in patients with malignant or immune conditions. Polymorphism analysis of the size of the third complementarity-determining region (CDR3) of heavy chain antibody transcripts was used to differentiate between a polyclonal infiltrate and a monoclonal infiltrate within a clinical vitreous sample of two groups of patients. PIOL was confirmed in all patients of the first group (n = 6). Five patients with autoimmune uveitis or immune-recovery uveitis associated with AIDS were included in the control group. The level of IL-10 in the vitreous was determined in all patients. In five cases of severe PIOL, CDR3 polymorphism analysis confirmed the presence of a dominant B-cell clone within the eye. In one case of confirmed PIOL presenting with mild vitritis, CDR3 polymorphism analysis was consistent with the existence of a polyclonal profile in the ocular sample studied. Conversely, it was shown that the detection of an intraocular monoclonal B-cell population does not necessarily imply ocular lymphoma. A clonal expansion was detected in a control patient who exhibited merely a nonmalignant response associated with immune-recovery uveitis. This PCR-based technique can make an important contribution to the characterization of intraocular B cells, but it alone cannot confirm or exclude the existence of a malignant lymphocyte proliferation. In the evaluation of a patient with intraocular inflammation in whom PIOL is suspected, CDR3 polymorphism analysis is recommended to confirm clonality. In general, the information about lymphocyte diversity provided by this technology opens up new possibilities for the analysis of ocular infiltrates.


Congenital anomalies of the spleen from an embryological point of view.
The spleen is the major accumulation of lymphoid tissue in the human body, an organ which prenatally produces and postnatally controls blood cells. Normally, a developed spleen lies in the upper left quadrant in parallel with the long axis of the 10th rib. It is a mesodermal derivate which first appears as a condensation of mesenchymal cells inside the dorsal mesogastrium at the end of the fourth embryonic week. Some congenital anomalies of the spleen are common, such as splenic lobulation and accessory spleen, while other conditions are rare, such as wandering spleen and polysplenia. Splenogonadal fusion is also a rare developmental anomaly, resulting from abnormal fusion of the splenic and gonadal primordia during prenatal development. The purpose of this article is to describe the normal development of the human spleen, supplemented with our own photomicrographs and a review of congenital anomalies of the spleen with their possible embryonic basis.


A Pragmatic Approach to Inpatient Diabetes Management during the COVID-19 Pandemic.
The pandemic of COVID-19 has presented new challenges to hospital personnel providing care for infected patients with diabetes who represent more than 20% of critically ill patients in intensive care units. Appropriate glycemic management contributes to a reduction in adverse clinical outcomes in acute illness but also requires intensive patient interactions for bedside glucose monitoring, intravenous and subcutaneous insulin administration, as well as rapid intervention for hypoglycemia events. These tasks are required at a time when minimizing patient interactions is recommended as a way of avoiding prolonged exposure to COVID-19 by health care personnel who often practice in settings with limited supplies of personal protective equipment. The purpose of this manuscript is to provide guidance for clinicians for reconciling recommended standards of care for infected hospitalized patients with diabetes while also addressing the daily realities of an overwhelmed health care system in many areas of the country. The use of modified protocols for insulin administration, bedside glucose monitoring, and medications such as glucocorticoids and hydroxychloroquine that may affect glycemic control are discussed. Continuous glucose monitoring systems have been proposed as an option for reducing time spent with patients, but there are important issues that need to be addressed if these are used in hospitalized patients. On-site and remote glucose management teams have potential to provide guidance in areas where there are shortages of personnel who have expertise in inpatient glycemic management.


Formononetin suppresses the proliferation of human non-small cell lung cancer through induction of cell cycle arrest and apoptosis.
Formononetin is a novel herbal isoflavonoid isolated from Astragalus membranaceus and possesses antitumorigenic properties. In the present study, we investigated the anti-proliferative effects of formononetin on human non-small cell lung cancer (NSCLC), and further elucidated the molecular mechanism underlying the anti-tumor property. MTT assay showed that formononetin treatment significantly inhibited the proliferation of two NSCLC cell lines including A549 and NCI-H23 in a time- and dose-dependent manner. Flow cytometric analysis demonstrated that formononetin induced G1-phase cell cycle arrest and promoted cell apoptosis in NSCLC cells. On the molecular level, we observed that exposure to formononetin altered the expression levels of cell cycle arrest-associated proteins p21, cyclin A and cyclin D1. Meanwhile, the apoptosis-related proteins cleaved caspase-3, bax and bcl-2 were also changed following treatment with formononetin. In addition, the expression level of p53 was dose-dependently upregulated after administration with formononetin. We also found that formononetin treatment increased the phosphorylation of p53 at Ser15 and Ser20 and enhances its transcriptional activity in a dose-dependent manner. Collectively, these results demonstrated that formononetin might be a potential chemopreventive drug for lung cancer therapy through induction of cell cycle arrest and apoptosis in NSCLC cells.


Pretreatment EGFR T790M mutation and BRCA1 mRNA expression in erlotinib-treated advanced non-small-cell lung cancer patients with EGFR mutations.
Advanced non-small-cell lung cancer (NSCLC) patients harboring epidermal growth factor receptor (EGFR) mutations (deletion in exon 19 or L858R) show an impressive progression-free survival of 14 months when treated with erlotinib. However, the presence of EGFR mutations can only imperfectly predict outcome. We hypothesized that progression-free survival could be influenced both by the pretreatment EGFR T790M mutation and by components of DNA repair pathways. We assessed the T790M mutation in pretreatment diagnostic specimens from 129 erlotinib-treated advanced NSCLC patients with EGFR mutations. The expression of eight genes and two proteins involved in DNA repair and four receptor tyrosine kinases was also examined. The EGFR T790M mutation was observed in 45 of 129 patients (35%). Progression-free survival was 12 months in patients with and 18 months in patients without the T790M mutation (P = 0.05). Progression-free survival was 27 months in patients with low BRCA1 mRNA levels, 18 months in those with intermediate levels, and 10 months in those with high levels (P = 0.02). In the multivariate analysis, the presence of the T790M mutation (HR, 4.35; P = 0.001), intermediate BRCA1 levels (HR, 8.19; P < 0.0001), and high BRCA1 levels (HR, 8.46; P < 0.0001) emerged as markers of shorter progression-free survival. Low BRCA1 levels neutralized the negative effect of the T790M mutation and were associated with longer progression-free survival to erlotinib. We advocate baseline assessment of the T790M mutation and BRCA1 expression to predict outcome and provide alternative individualized treatment to patients based on T790M mutations and BRCA1 expression.


Diagnostic Biomarkers of Alzheimer's Disease as Identified in Saliva using 1H NMR-Based Metabolomics.
Using 1H NMR metabolomics, we biochemically profiled saliva samples collected from healthy-controls (n = 12), mild cognitive impairment (MCI) sufferers (n = 8), and Alzheimer's disease (AD) patients (n = 9). We accurately identified significant concentration changes in 22 metabolites in the saliva of MCI and AD patients compared to controls. This pilot study demonstrates the potential for using metabolomics and saliva for the early diagnosis of AD. Given the ease and convenience of collecting saliva, the development of accurate and sensitive salivary biomarkers would be ideal for screening those at greatest risk of developing AD.


Comparison of computational methods to model DNA minor groove binders.
There has been a profound interest in designing small molecules that interact in sequence-selective fashion with DNA minor grooves. However, most in silico approaches have not been parametrized for DNA ligand interaction. In this regard, a systematic computational analysis of 57 available PDB structures of noncovalent DNA minor groove binders has been undertaken. The study starts with a rigorous benchmarking of GOLD, GLIDE, CDOCKER, and AUTODOCK docking protocols followed by developing QSSR models and finally molecular dynamics simulations. In GOLD and GLIDE, the orientation of the best score pose is closer to the lowest rmsd pose, and the deviation in the conformation of various poses is also smaller compared to other docking protocols. Efficient QSSR models were developed with constitutional, topological, and quantum chemical descriptors on the basis of B3LYP/6-31G* optimized geometries, and with this ΔT(m) values of 46 ligands were predicted. Molecular dynamics simulations of the 14 DNA-ligand complexes with Amber 8.0 show that the complexes are stable in aqueous conditions and do not undergo noticeable fluctuations during the 5 ns production run, with respect to their initial placement in the minor groove region.


Oral contraceptives, androgens, and the sexuality of young women: II. The role of androgens.
The relationships between plasma free testosterone (FT) and measures of sexual attitude, sexual behavior, and gender role behavior were assessed in 55 oral contraceptive-using and 53 nonusing female undergraduates. Plasma FT and other measures of androgenicity were substantially lower in the oral contraceptive (OC) group. Correlations between FT and certain behavioral and attitudinal measures were found in the OC users but not the nonusers. In the OC users, FT was positively associated with frequency of sexual intercourse but not with frequency of masturbation. It was negatively associated with restrictive sexual morality. Correlations between FT and measures of gender role behavior were negligible, and FT was unrelated to proceptivity, homosexual interest, or the use of sexual fantasy. The occurrence of some predicted correlations among pill-using women but not the nonusers requires explanation, particularly in view of the substantially lower levels of FT in the pill-using group. It is suggested that androgen-behavior relationships in women are easily obscured by psychosocial influences and in this sample of young women such influences may have been more powerful among those not using OCs. Such psychosocial influences are likely to differ at different stages of women's life cycles. The importance of controlling for such influences in any study of hormone-sexual behavior relationships in women is emphasized, and the need for prospective studies of women before and after starting on steroidal contraception is recognized.


Syngeneic thyroglobulin is immunogenic in good responder mice.
Mouse thyroglobulin (MTg) or thyroid extract (TE) was given repeatedly to good responder C3H/Anf (H-2k) and poor responder BALB/c (H-2d) mice in the absence of adjuvant. Anti-MTg antibodies reached high levels in good responder mice given high doses of thyroid antigen. To eliminate stimulation by alloantigenic determinants and reduce the chance of denaturation. TE from syngeneic mice was prepared freshly each week and injected into good and poor responder strains. Again, significant antibody titers were observed in good responder mice. The antibody was specific for MTg since (a) it was not inhibited by extracts of other organs and (b) it reacted strongly with the closely related rat Tg and weakly with Tg from other species. Histology revealed mononuclear cell infiltration of the thyroid of good responder, but not of poor responder mice, regardless of the strain used to provide the thyroid antigen. The data demonstrate the presence of both T and B cell populations reactive with this self antigen and their stimulation by repeated high doses of antigen, without the aid of adjuvant, to override the regulatory controls that normally prevent autoimmune responses.


Impact of hospital case volume on testicular cancer outcomes and practice patterns.
Given the rarity of testicular germ cell tumors (TGCTs) and the complex aspects of management, we evaluate the effect of hospital TGCT case volume on overall survival outcomes and practice patterns. The National Cancer Database was queried for patients diagnosed with seminoma or nonseminomatous germ cell tumor (NSGCT). Hospitals were classified by case volume as high (99th percentile, ≥26.1 cases annually), high-intermediate (95-99th percentile, 14.6-26.0 cases annually), intermediate (75-95th percentile, 6.1-14.5 cases annually), low-intermediate (25-75th percentile, 1.8-6.0 cases annually), and low (25th percentile,<1.8 cases annually). The median (interquartile range) number of TGCT cases per institution per year was 3.4 (1.8-6.1). A total of 33,417 patients with TGCT diagnosed from 1,239 institutions met inclusion criteria. Despite worse disease characteristics of patients treated at higher volume institutions, hospital volume was positively associated with survival outcomes in more advanced cases of TGCT. In the overall cohort, compared to the high-volume hospitals, patients treated at high-intermediate, intermediate, low-intermediate, and low volume hospitals the hazard ratio for overall mortality was 1.28, 1.45, 1.48, and 1.83, respectively (P<0.05). The association between survival and hospital volume was not apparent for seminoma or stage I NSGCT. Patients treated at higher volume hospitals were more likely to undergo surveillance for stage I seminoma, primary retroperitoneal lymph node dissection (RPLND) for stage I NSGCT, and postchemotherapy RPLND for stage II/III NSGCT. Our analysis of a nationwide cancer registry demonstrated that increased hospital TGCT case volume was associated with significant differences in management strategies and improved survival outcomes, in particular for more advanced disease.


Preventing early childhood transmission of hepatitis B in remote aboriginal communities in Northern Australia.
Chronic hepatitis B is a public health concern in Aboriginal communities in the Northern Territory of Australia with prevalence almost four times the non-Aboriginal population. Infection is suspected to mainly occur in early life, however, the mode of transmission and vaccine effectiveness is not known in this population. WHO has set a target for hepatitis B elimination by 2030; elimination in this disproportionately affected population in Australia will require understanding of the modes of transmission and vaccine effectiveness. We conducted the study at four very remote Aboriginal communities. We approached mothers who had chronic hepatitis B and had given birth between 1988 and 2013 for consent. We obtained hepatitis B serology, immunisation and birth details from the medical record. If both mother and child had hepatitis B viral DNA detected, we performed viral whole genome sequencing. We approached 45 women for consent, of whom 23 agreed to participate. We included 20 mothers and 38 of their children. Of the 20 included mothers, 5 (25%) had children who were hepatitis B immune by exposure and 3 (15%) had children with evidence of chronic hepatitis B infection at the time of assessment. Hepatitis B immunoglobulin (HBIg) had been given at birth in 29/38 (76.3, 95% CI 59.8-88.6) children, and 26 children (68.4, 95% CI 51.3-82.5) were fully vaccinated. Of the 3 children who had chronic hepatitis B, all had received HBIg at birth and two were fully vaccinated. Of the 5 who were immune by exposure, 4 had received HBIg at birth and one was fully vaccinated. Whole genome sequencing revealed one episode of definite mother to child transmission. There was also one definite case of horizontal transmission. Chronic hepatitis B in this context is a sensitive issue, with a high proportion of women refusing consent. Although uncommon, there is ongoing transmission of hepatitis B to Aboriginal children in remote northern Australia despite vaccination, and this is likely occurring by both vertical and horizontal routes. Prevention will require ongoing investment to overcome the many barriers experienced by this population in accessing care.


Fentanyl-Induced Chest Wall Rigidity as a Cause of Acute Respiratory Failure in the Intensive Care Unit.
We aim to report a case of chest wall rigidity induced by high-dose fentanyl infusion sedation and analgesia in the intensive care unit (ICU) for management of pneumonia and asthma. The patient is an 80-year-old woman, who presented to the hospital with complaints of fever and productive cough with yellowish expectoration of 2 days duration. She also had lethargy over the same time period and had sick contacts in the form of two daughters who both had recently recovered from a "flu-like" illness. She was known to have bronchial asthma treated with seretide 25/250 two puffs ON + PO monteleukast 10 mg ON, hypertension treated with PO losartan 50 mg BD, type 2 diabetes mellitus controlled with PO linagliptin 5 mg OM and a previous right thalamic ischemic stroke 5 years ago for which she was on PO clopidogrel 75 mg OM and PO simvastatin. She developed severe ventilator desynchrony characterized by dramatic sudden onset of severe hypercarbia, severely decreased pulmonary compliance and episodic breath holding. She was empirically treated for asthma exacerbation and treated with steroids, bronchodilators and manual ventilation but despite doing so during this episode the patient failed to respond. The patient was clinically evaluated and dynamic hyperinflation was excluded as a cause of the respiratory failure. There was no evidence of pneumothorax and worsening pneumonia. Considerations of insufficient sedation and analgesia led to deepening sedation and analgesia without good response. Ventilation was dramatically improved after small doses of neuromuscular relaxation. This cycle was repeated many times. The patient was kept on high-dose propofol and fentanyl, but there were repeated cycles of sudden persistent severe hypercarbia, severely decreased pulmonary compliance and episodic breath holding. Eventually a suspicion of fentanyl-induced chest wall rigidity was made after excluding causes of airway resistance and reduction in pulmonary compliance. Gradual reduction in fentanyl infusion was associated with a reduction of episodes of reduced lung compliance and improvement in ventilation. Fentanyl is often used for analgesia and sedation in the ICU. It has a good side effect profile but it is not without harm. High doses of fentanyl can lead to dramatic worsening of respiratory mechanics that may be life threatening. Fentanyl-induced chest wall rigidity is an important side effect that needs to be considered in the differential diagnosis of respiratory failure in the ICU.


Clinical implication of voltage-dependent anion channel 1 in uterine cervical cancer and its action on cervical cancer cells.
Two-dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometry were performed to investigate the influence of human nonmetastatic clone 23 type 1 (nm23-H1), a metastasis-associated gene on proteomic alterations in cancer cells of the uterine cervix. It was validated by RT-PCR and Western blot analysis. The expression of voltage-dependent anion channel 1 (VDAC1) was increased in nm23-H1 gene silenced SiHa or CaSki cervical cancer cells. The clinical implication was shown that cervical cancer tissues with positive VDAC1 immunoreactivity exhibited deep stromal invasion (>10 mm in depth) and large tumor size (> 4 cm in diameter). Cervical cancer patients with positive VDAC1 immunoreactivity displayed higher recurrence and poorer overall survival than those with negative VDAC1. Silencing of VDAC1 reduced cell proliferation and migratory ability. Mitochondrial membrane potential was decreased and reactive oxygen species generation was increased in the VDAC1 gene-silenced cervical cancer cells. Cell cycle progression and autophagy were not changed in VDAC1 silencing cells. The cytotoxicity of cisplatin was significantly enhanced by knockdown of cellular VDAC1 and the compounds that interfere with hexokinase binding to VDAC. Therapeutic strategies may be offered using VDAC1 as a target to reduce cell growth and migration, enhance the synergistic therapeutic efficacy of cisplatin and reduce cisplatin dose-limiting toxicity.


Results of a survey of diagnosed HIV infections prevalent in 1996 in England and Wales.
This paper describes a survey undertaken to assess the caseload of HIV infected patients who received medical care from statutory service providers in England and Wales in 1996 in order to inform health authorities about the size and composition of their resident population of such patients. A total of 13,670 HIV infected patients were identified as living in England and Wales, 70% of whom lived in the Thames NHS executive regions. Over half the total caseload received care within their health authority of residence. Regional care centres attracted patients from wider areas, however, particularly in the North West and Thames regions. This survey of prevalent diagnosed HIV infections, one of a series conducted annually, provides public health specialists with information relevant to their localities without compromising patient confidentiality. Along with other data from the surveillance of AIDS cases and HIV infections it contributes to the assessment and projection of demands on health and social services and provides evidence on which to develop and direct national and local health campaigns.


Time-dependent activity and expression of glutathione S-transferases in the human colon adenocarcinoma cell line Caco-2.
The human colon carcinoma cell line Caco-2 was examined for glutathione S-transferase (GST) composition and activity. Freshly seeded cells and cells until 4 days after confluency contain only the placental (Pi) form of glutathione transferase. Cells in culture for longer periods start to express class-Alpha GST isoenzymes. Confluent cells in culture for 20 days or longer contain up to 90% class-Alpha GST. Class-Mu GSTs are not detectable. GST activity gradually increases from 564 +/- 28 to 5381 +/- 165 nmol/min per mg of protein at day 0 and 32 after confluency respectively. With regard to GST composition, Caco-2 cells in culture for longer periods most resemble small-intestinal cells, whereas short-time cultures have characteristics of colonic cells. This cell line is very well suited for the study of both the in vitro properties and the expression of class Alpha and Pi GSTs.


The pattern of surgically treatable anorectal diseases in University of Port Harcourt Teaching Hospital, Rivers State, Nigeria.
Disorders of the anus and rectum occurcommonly in surgical practice. Common anorectal disorders include haemorrhoids, carcinoma of the rectum, anal fissures, anorectal fistulae and abscesses. Less frequently encountered disorders include perineal injuries, anal cancers and rectal polyps. The purpose of this study is to determine the pattern of anorectal diseases which were surgically treated in University of Port Harcourt Teaching Hospital. This is a 4 year retrospective study of all adult patients with anorectal diseases who were admitted into the surgical wards of University of Port Harcourt Teaching Hospital. Relevant data were retrieved and analyzed. One hundred and fifty cases were seen over the 4 year period. There were 92 (61.33%) males and 58 (38.67%) females making a male to female ratio of 1.6:1. The peak age incidence was 31-40 years. The commonest anorectal disorder was haemorrhoids. Other commonly encountered conditions in decreasing order were carcinoma of the rectum, anorectal fistulae, anal fissures and rectal prolapsed. Five patients died and they all had carcinoma of the rectum. Haemorrhoids remain the commonest anorectal disorder in University of Port Harcourt Teaching Hospital. Carcinoma of the rectum which was not noted among the studied population in ourcentre 10 years ago now assumes the second position and this reflects an increasing incidence of the disease. Therefore, early diagnosis and treatment are imperative for increase in survival rate.


Nonstructural Protein 2 Is Critical to Infection Efficiency of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus on PAMs and Influence Virulence In Vivo.
Porcine reproductive and respiratory syndrome (PRRS) is an important viral disease, causing significant economic losses to the swine industry worldwide. Atypical cases caused by highly pathogenic PRRS virus (HP-PRRSV) emerged in 2006 in China. The vaccine strain HuN4-F112 has been developed from the wild-type HP-PRRSV HuN4 through repeated passages on MARC-145 cells. However, the mechanisms of attenuation have yet to be defined. Previous studies have shown that the vaccine strain HuN4-F112 could not effectively replicate in porcine alveolar macrophages (PAMs). In the present study, a series of chimeric and mutant PRRSVs were constructed to investigate regions associated with the virus attenuation. Firstly, the corresponding genome regions (ORF1a, ORF1b and ORFs 2-7) were exchanged between two infectious clones of HuN4 and HuN4-F112, and then the influence of small regions in ORF1a and ORF2-7 was evaluated, then influence of specific amino acids on NSP2 was tested. NSP2 was determined to be the key gene that regulated infection efficiency on PAMs, and amino acids at 893 and 979 of NSP2 were the key amino acids. The results of in vivo study indicated that NSP2 was not only important for infection efficiency in vitro, but also influenced the virulence, which was indicated by the results of survival rate, temperature, viremia, lung score and tissue score.