Genome-wide analysis of UDP-glycosyltransferase gene family and identification of a flavonoid 7-O-UGT (AhUGT75A) enhancing abiotic stress in peanut (Arachis hypogaea L.).
Glycosylation, catalyzed by UDP-glycosyltransferase (UGT), was important for enhancing solubility, bioactivity, and diversity of flavonoids. Peanut (Arachis hypogaea L.) is an important oilseed and cash crop worldwide. In addition to provide high quality of edible oils and proteins, peanut seeds contain a rich source of flavonoid glycosides that benefit human health. However, information of UGT gene family was quite limited in peanut. In present study, a total of 267 AhUGTs clustered into 15 phylogenetic groups were identified in peanut genome. Group I has greatly expanded to contain the largest number of AhUGT genes. Segmental duplication was the major driving force for AhUGT gene family expansion. Transcriptomic analysis of gene expression profiles in various tissues and under different abiotic stress treatments indicated AhUGTs were involved in peanut growth and abiotic stress response. AhUGT75A (UGT73CG33), located in mitochondria, was characterized as a flavonoid 7-O-UGT by in vitro enzyme assays. The transcript level of AhUGT75A was strongly induced by abiotic stress. Overexpression of AhUGT75A resulted in accumulating less amount of malondialdehyde (MDA) and superoxide, and enhancing tolerance against drought and/or salt stress in transgenic Arabidopsis. These results indicated AhUGT75A played important roles in conferring abiotic stress tolerance through reactive oxygen species scavenging. Our research only not provides valuable information for functional characterization of UGTs in peanut, but also gives new insights into potential applications in breeding new cultivars with both desirable stress tolerance and health benefits.


[Pediatric aspects of osteoporosis].
The skeleton is not a single entity; the age of onset, rate of accrual and age of attainment of peak bone density vary according to gender and according to region. Exposure of life style factors during growth and disease may produce site specific deficits in bone density in adulthood. Since bone mass in the elderly depends beside the rate of involutional bone loss mainly on the peak bone mass its optimal development in childhood and adolescence appears as being the key issue problem in prophylactic strategy of osteoporotic therapy.


Cardiovascular alterations at different stages of hypertension development during ethanol consumption: time-course of vascular and autonomic changes.
The aim of the present work was to establish a time-course correlation between vascular and autonomic changes that contribute to the development of hypertension during ethanol ingestion in rats. For this, male Wistar rats were subjected to the intake of increasing ethanol concentrations in their drinking water during four weeks. Ethanol effects were investigated at the end of each week. Mild hypertension was already observed at the first week of treatment, and a progressive blood pressure increase was observed along the evaluation period. Increased pressor response to phenylephrine was observed from first to fourth week. α1-Adrenoceptor protein in the mesenteric bed was enhanced at the first week, whereas β2-adrenoceptor protein in the aorta was reduced after the second week. In the third week, ethanol intake facilitated the depressor response to sodium nitroprusside, whereas in the fourth week it reduced nitrate content in aorta and increased it plasma. The bradycardic component of the baroreflex was impaired, whereas baroreflex tachycardia was enhanced at the third and fourth weeks. AT1A receptor and C-type natriuretic peptide (CNP) mRNAs in the nucleus tractus solitarius were increased at the fourth week. These findings suggest that increased vascular responsiveness to vasoconstrictor agents is possibly a link factor in the development and maintenance of the progressive hypertension induced by ethanol consumption. Additionally, baroreflex changes are possibly mediated by alterations in angiotensinergic mechanisms and CNP content within the brainstem, which contribute to maintaining the hypertensive state in later phases of ethanol ingestion. Facilitated vascular responsiveness to nitric oxide seems to counteract ethanol-induced hypertension.


Neutrophil/Lymphocyte ratio is associated with non-calcified plaque burden in patients with coronary artery disease.
Elevations in soluble markers of inflammation and changes in leukocyte subset distribution are frequently reported in patients with coronary artery disease (CAD). Lately, the neutrophil/lymphocyte ratio has emerged as a potential marker of both CAD severity and cardiovascular prognosis. The aim of the study was to investigate whether neutrophil/lymphocyte ratio and other immune-inflammatory markers were related to plaque burden, as assessed by coronary computed tomography angiography (CCTA), in patients with CAD. Twenty patients with non-ST-elevation acute coronary syndrome (NSTE-ACS) and 30 patients with stable angina (SA) underwent CCTA at two occasions, immediately prior to coronary angiography and after three months. Atherosclerotic plaques were classified as calcified, mixed and non-calcified. Blood samples were drawn at both occasions. Leukocyte subsets were analyzed by white blood cell differential counts and flow cytometry. Levels of C-reactive protein (CRP) and interleukin(IL)-6 were measured in plasma. Blood analyses were also performed in 37 healthy controls. Plaque variables did not change over 3 months, total plaque burden being similar in NSTE-ACS and SA. However, non-calcified/total plaque ratio was higher in NSTE-ACS, 0.25(0.09-0.44) vs 0.11(0.00-0.25), p<0.05. At admission, levels of monocytes, neutrophils, neutrophil/lymphocyte ratios, CD4+ T cells, CRP and IL-6 were significantly elevated, while levels of NK cells were reduced, in both patient groups as compared to controls. After 3 months, levels of monocytes, neutrophils, neutrophil/lymphocyte ratios and CD4+ T cells remained elevated in patients. Neutrophil/lymphocyte ratios and neutrophil counts correlated significantly with numbers of non-calcified plaques and also with non-calcified/total plaque ratio (r = 0.403, p = 0.010 and r = 0.382, p = 0.024, respectively), but not with total plaque burden. Among immune-inflammatory markers in NSTE-ACS and SA patients, neutrophil counts and neutrophil/lymphocyte ratios were significantly correlated with non-calcified plaques. Data suggest that these easily measured biomarkers reflect the burden of vulnerable plaques in CAD.


Cerebral salt wasting in a child with cervicothoracic hematoma.
Although the syndrome of inappropriate antidiuretic hormone secretion (SIADH) is commonly observed in patients with acute or chronic central nervous system (CNS) disorders, cerebral salt wasting (CSW) that results in hyponatremia has rarely been reported in children. Both SIADH and CSW result in increased urinary sodium excretion and hyponatremia. However, the management protocols for these two conditions are quite different; volume restriction is used in treating SIADH, while volume expansion is necessary for the treatment of CSW. We present a case of CSW in a child with cervicothoracic hematoma secondary to head and cervicothoracic trauma, without evidence of brain edema. The child was diagnosed on the basis of high urinary sodium excretion resulting in hyponatremia and low serum osmolarity. Improvements in serum sodium levels after saline hydration confirmed this diagnosis. We believe that potentially dangerous cases of hyponatremia should be carefully evaluated in children with cervicothoracic hematoma secondary to trauma, including situations in which brain edema is absent.


VBAC: antenatal predictors of success.
To determine antenatal factors that may predict successful vaginal birth after Caesarean section (VBAC), to develop a relevant antenatal scoring system and a nomogram for prediction of vaginal birth after caesarean delivery. A non recurring indication for previous Caesarean section (CS), such as breech presentation or foetal distress, is associated with a much higher successful VBAC rate than recurrent indications, such as cephalopelvic disproportion (CPD). Prior vaginal deliveries are excellent prognostic indicators of successful VBAC, especially if the vaginal delivery follows the prior CS. A low vertical uterine incision does not seem to adversely affect VBAC success rates as compared to a low transverse incision. Maternal obesity and diabetes mellitus adversely affect VBAC outcomes. Foetal macrosomia does not appear to be a contraindication to VBAC, as success rates exceeding 50% are achieved and uterine rupture rates are not increased. An inter-pregnancy interval of <24 months is not associated with a decreased success of VBAC. Success rates decrease when interval increases. Twin gestation does not preclude VBAC. Post-dates pregnancies may deliver successfully by VBAC in greater than two-thirds of cases. There are few absolute contraindications to attempted VBAC. Attempted VBAC will be successful in the majority of attempted cases.


Contribution to the neurohaemal organs in pterygote insects.
In this review a comprehensive account of anatomy and various cell types occurring in the neurohaemal organs in Pterygote insects has been given. The chemical nature and composition of neurohaemal secretions and the mechanisms of liberation of neurosecretory material (NSM) from neurohaemal organs (NHO) have been described. Also the effects of neurhaemal secretions on diuresis, feeding and growth, heart beat, pigmentation of cuticle, carbohydrate metabolism, oocyte development and respiratory metabolism have been incorporated.


Reconstructing adhesion structures in tissues by cryo-electron tomography of vitrified frozen sections.
Cryo-electron tomography enables three-dimensional insights into the macromolecular architecture of cells in a close-to-life state. However, it is limited to thin specimens, <1.0 μm in thickness, typically restricted to the peripheral areas of intact eukaryotic cells. Analysis of tissue ultrastructure, on the other hand, requires physical sectioning approaches, preferably cryo-sectioning, following which electron tomography (ET) may be performed. Nevertheless, cryo-electron microscopy of vitrified sections is a demanding technique and typically cannot be used to examine thick sections, >80-100 nm, due to surface crevasses. Here, we explore the potential use of cryo-ET of vitrified frozen sections (VFSs) for imaging cell adhesions in chicken smooth muscle and mouse epithelial tissues. By investigating 300-400 nm thick sections, which are collected on the EM grid and re-vitrified, we resolved fine 3D structural details of the membrane-associated dense plaques and flanking caveoli in smooth muscle tissue, and desmosomal adhesions in stratified epithelium. Technically, this method offers a simple approach for reconstructing thick volumes of hydrated frozen sections.


[Regulatory selection as an alternative to the neutrality theory].
Regulatory selection is the most typical mode of natural selection responsible for the optimization of the regulatory systems of an organism. In addition to regulatory selection, stringent selection of pleiotropic mutants or recombinant forms with altered ecological potential operates in successful species during vertical evolution. Regulatory selection stabilizes alterations in the ecological potential, acting at intervals between larger regulatory genome alterations occurring at the boundaries between evolutionary phases. Fisher's adaptive model can be applied to regulatory selection. At each separate evolutionary phase, the selection coefficients in selected mutants decline gradually. At the final stage of the evolutionary phase, mutations with a minor selective advantage are selected. Hence, regulatory selection may be regarded as an alternative to the neutral evolution theory.


Survival and prognostic factors in 212 Italian patients with genetic hemochromatosis.
Two hundred twelve Italian patients with genetic hemochromatosis (181 men, mean age 50 +/- 11 yr; and 31 women, mean age 49 +/- 10 yr) were followed for a median period of 44 mo (range = 3 to 218 mo). Alcohol abuse was present in 31 subjects (15%), and chronic HBV and HCV infection were seen in 19 (9%) and 35 (24%) of 145 cases tested, respectively. Twenty-four patients (11%) had concomitant beta-thalassemia trait. Liver biopsy revealed cirrhosis in 146 and a noncirrhotic pattern in the other 66. Perls' stain was degree III in 37 patients and IV in 171 patients. One hundred eighty-five patients underwent weekly venesection, and iron depletion was achieved in 122 cases after total iron removal of 3 to 41 gm. Death occurred in 44 patients after 3 to 198 mo and was due to hepatocellular carcinoma in 20 cases, liver failure in 10, extrahepatic cancer in six, heart failure in three and hemochromatosis unrelated causes in five. Cancer has developed in seven other patients still alive (hepatocellular in five and extrahepatic in two). No deaths were observed among noncirrhotic patients; cumulative survival rates in cirrhotic patients were 85%, 75%, 60% and 47% at 3, 5, 8 and 10 yr, respectively. Univariate analysis in the 146 cirrhotic patients showed that age greater than 60 yr, alcohol abuse, cardiomyopathy, skin pigmentation, portal hypertension, hypoalbuminemia, hypergammaglobulinemia and Child class B or C had significant negative prognostic value. At multivariate analysis, only alcohol abuse, gamma-globulins greater than 2.0 gm/dl and Child class B or C maintained their negative prognostic values (p less than 0.01, hazard ratio 2.7; p less than 0.001, hazard ratio 2.8; and p less than 0.001, hazard ratio 4.3, respectively).


Proper regulation of a sperm-specific cis-nat-siRNA is essential for double fertilization in Arabidopsis.
Natural cis-antisense siRNAs (cis-nat-siRNAs) are a recently characterized class of small regulatory RNAs that are widespread in eukaryotes. Despite their abundance, the importance of their regulatory activity is largely unknown. The only functional role for eukaryotic cis-nat-siRNAs that has been described to date is in environmental stress responses in plants. Here we demonstrate that cis-nat-siRNA-based regulation plays key roles in Arabidopsis reproductive function, as it facilitates gametophyte formation and double fertilization, a developmental process of enormous agricultural value. We show that male gametophytic kokopelli (kpl) mutants display frequent single-fertilization events, and that KPL and a inversely transcribed gene, ARIADNE14 (ARI14), which encodes a putative ubiquitin E3 ligase, generate a sperm-specific nat-siRNA pair. In the absence of KPL, ARI14 RNA levels in sperm are increased and fertilization is impaired. Furthermore, ARI14 transcripts accumulate in several siRNA biogenesis pathway mutants, and overexpression of ARI14 in sperm phenocopies the reduced seed set of the kokopelli mutants. These results extend the regulatory capacity of cis-nat-siRNAs to development by identifying a role for cis-nat-siRNAs in controlling sperm function during double fertilization.


The Cerberus/Dan-family protein Charon is a negative regulator of Nodal signaling during left-right patterning in zebrafish.
We have isolated a novel gene, charon, that encodes a member of the Cerberus/Dan family of secreted factors. In zebrafish, Fugu and flounder, charon is expressed in regions embracing Kupffer's vesicle, which is considered to be the teleost fish equivalent to the region of the mouse definitive node that is required for left-right (L/R) patterning. Misexpression of Charon elicited phenotypes similar to those of mutant embryos defective in Nodal signaling or embryos overexpressing Antivin(Atv)/Lefty1, an inhibitor for Nodal and Activin. Charon also suppressed the dorsalizing activity of all three of the known zebrafish Nodal-related proteins (Cyclops, Squint and Southpaw), indicating that Charon can antagonize Nodal signaling. Because Southpaw functions in the L/R patterning of lateral plate mesoderm and the diencephalon, we asked whether Charon is involved in regulating L/R asymmetry. Inhibition of Charon's function by antisense morpholino oligonucleotides (MOs) led to a loss of L/R polarity, as evidenced by bilateral expression of the left side-specific genes in the lateral plate mesoderm (southpaw, cyclops, atv/lefty1, lefty2 and pitx2) and diencephalon (cyclops, atv/lefty1 and pitx2), and defects in early (heart jogging) and late (heart looping) asymmetric heart development, but did not disturb the notochord development or the atv/lefty1-mediated midline barrier function. MO-mediated inhibition of both Charon and Southpaw led to a reduction in or loss of the expression of the left side-specific genes, suggesting that Southpaw is epistatic to Charon in left-side formation. These data indicate that antagonistic interactions between Charon and Nodal (Southpaw), which take place in regions adjacent to Kupffer's vesicle, play an important role in L/R patterning in zebrafish.


Higher plane of nutrition pre-weaning enhances Holstein calf mammary gland development through alterations in the parenchyma and fat pad transcriptome.
To reduce costs of rearing replacement heifers, researchers have focused on decreasing age at breeding and first calving. To increase returns upon initiation of lactation the focus has been on increasing mammary development prior to onset of first lactation. Enhanced plane of nutrition pre-weaning may benefit the entire replacement heifer operation by promoting mammary gland development and greater future production. Twelve Holstein heifer calves (< 1 week old) were reared on 1 of 2 dietary treatments (n = 6/group) for 8 weeks: a control group fed a restricted milk replacer at 0.45 kg/d (R, 20% crude protein, 20% fat), or an accelerated group fed an enhanced milk replacer at 1.13 kg/d (EH, 28% crude protein, 25% fat). At weaning (8 weeks), calves were euthanized and sub-samples of mammary parenchyma (PAR) and mammary fat pad (MFP) were harvested upon removal from the body. Total RNA from both tissues was extracted and sequenced using the Illumina HiSeq2500 platform. The Dynamic Impact Approach (DIA) and Ingenuity Pathway Analysis (IPA) were used for pathway analysis and functions, gene networks, and cross-talk analyses of the two tissues. When comparing EH vs R 1561 genes (895 upregulated, 666 downregulated) and 970 genes (506 upregulated, 464 downregulated) were differentially expressed in PAR and MFP, respectively. DIA and IPA results highlight a greater proliferation and differentiation activity in both PAR and MFP, supported by an increased metabolic activity. When calves were fed EH, the PAR displayed transcriptional signs of greater overall organ development, with higher ductal growth and branching, together with a supportive blood vessel and nerve network. These activities were mediated by intracellular cascades, such as AKT, SHH, MAPK, and Wnt, probably activated by hormones, growth factors, and endogenous molecules. The analysis also revealed strong communication between MFP and PAR. The transcriptomics and bioinformatics approach highlighted key mechanisms that mediate the mammary gland response to a higher plane of nutrition in the pre-weaning period.


Cassava brown streak virus Ham1 protein hydrolyses mutagenic nucleotides and is a necrosis determinant.
Cassava brown streak disease (CBSD) is a leading cause of cassava losses in East and Central Africa, and is currently having a severe impact on food security. The disease is caused by two viruses within the Potyviridae family: Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV), which both encode atypical Ham1 proteins with highly conserved inosine triphosphate (ITP) pyrophosphohydrolase (ITPase) domains. ITPase proteins are widely encoded by plant, animal, and archaea. They selectively hydrolyse mutagenic nucleotide triphosphates to prevent their incorporation into nucleic acid and thereby function to reduce mutation rates. It has previously been hypothesized that U/CBSVs encode Ham1 proteins with ITPase activity to reduce viral mutation rates during infection. In this study, we investigate the potential roles of U/CBSV Ham1 proteins. We show that both CBSV and UCBSV Ham1 proteins have ITPase activities through in vitro enzyme assays. Deep-sequencing experiments found no evidence of the U/CBSV Ham1 proteins providing mutagenic protection during infections of Nicotiana hosts. Manipulations of the CBSV_Tanza infectious clone were performed, including a Ham1 deletion, ITPase point mutations, and UCBSV Ham1 chimera. Unlike severely necrotic wild-type CBSV_Tanza infections, infections of Nicotiana benthamiana with the manipulated CBSV infectious clones do not develop necrosis, indicating that that the CBSV Ham1 is a necrosis determinant. We propose that the presence of U/CBSV Ham1 proteins with highly conserved ITPase motifs indicates that they serve highly selectable functions during infections of cassava and may represent a euphorbia host adaptation that could be targeted in antiviral strategies.


Waldenström macroglobulinemia: clinical and immunological aspects, natural history, cell of origin, and emerging mouse models.
Waldenström macroglobulinemia (WM) is a rare and currently incurable neoplasm of IgM-expressing B-lymphocytes that is characterized by the occurrence of a monoclonal IgM (mIgM) paraprotein in blood serum and the infiltration of the hematopoietic bone marrow with malignant lymphoplasmacytic cells. The symptoms of patients with WM can be attributed to the extent and tissue sites of tumor cell infiltration and the magnitude and immunological specificity of the paraprotein. WM presents fascinating clues on neoplastic B-cell development, including the recent discovery of a specific gain-of-function mutation in the MYD88 adapter protein. This not only provides an intriguing link to new findings that natural effector IgM(+)IgD(+) memory B-cells are dependent on MYD88 signaling, but also supports the hypothesis that WM derives from primitive, innate-like B-cells, such as marginal zone and B1 B-cells. Following a brief review of the clinical aspects and natural history of WM, this review discusses the thorny issue of WM's cell of origin in greater depth. Also included are emerging, genetically engineered mouse models of human WM that may enhance our understanding of the biologic and genetic underpinnings of the disease and facilitate the design and testing of new approaches to treat and prevent WM more effectively.


Modeling stroke in mice: permanent coagulation of the distal middle cerebral artery.
Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical options are available for a small proportion of stroke patients in the acute phase. Current research is intensively searching for novel therapeutic strategies and is increasingly focusing on the sub-acute and chronic phase after stroke because more patients might be eligible for therapeutic interventions in a prolonged time window. These delayed mechanisms include important pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity and regeneration. In order to analyze these mechanisms and to subsequently evaluate novel drug targets, experimental stroke models with clinical relevance, low mortality and high reproducibility are sought after. Moreover, mice are the smallest mammals in which a focal stroke lesion can be induced and for which a broad spectrum of transgenic models are available. Therefore, we describe here the mouse model of transcranial, permanent coagulation of the middle cerebral artery via electrocoagulation distal of the lenticulostriatal arteries, the so-called "coagulation model". The resulting infarct in this model is located mainly in the cortex; the relative infarct volume in relation to brain size corresponds to the majority of human strokes. Moreover, the model fulfills the above-mentioned criteria of reproducibility and low mortality. In this video we demonstrate the surgical methods of stroke induction in the "coagulation model" and report histological and functional analysis tools.


A developmental exploration of language used to accompany symbolic play in young, normal children (2-4 years old).
Observation of play can provide crucial information about a child's developmental level of language. To make this information available categorization and measurement are necessary. By defining, from a cognitive and pragmatic point of view, separate but parallel measures of play and language it was possible to define four different levels of symbolic functioning: representational play; conceptual play; programmatic play; and script play. Careful longitudinal descriptions of language used to accompany symbolic play of ten 2-4-year-old children are presented. Interesting developmental trends in play and cognitive and pragmatic organization of language were found. Gradually language production was considered as the primary mode for symbolic expression between the ages of 2;6 (2 years; 6 months) and 3 years. Language production increased dramatically between the ages of 3 and 3;6, and became better differentiated from the play context. The scientific study of child language using an analytic tool, such as a play procedure, to describe and analyse spontaneous production of spoken language, with a cognitive and pragmatic framework contributes not only a more accurate understanding of normal play and language development, but may also have an efficient clinical value. Suggestions for and implications of exploring components of language used in a standardized play situation are discussed in reference to developmentally disabled children.


Fertility Intentions and Their Realization in Couples: How the Division of Household Chores Matters.
Most research on Europe indicates that a gender-balanced division of family work tends to increase childbearing probabilities, but empirical results vary substantially. The present article proposes explanations for this observed discrepancy. It develops prior research further by (1) studying short-term fertility intentions and their realization within the subsequent 4 years, (2) analyzing the role of the spouses' satisfaction with the division for the effects that the division may have on childbearing, (3) proving a mediation by relationship satisfaction, and (4) considering gender as well as parity as moderators. Using data from two waves of the Generations and Gender Survey, we show that the division of work affects childbearing intentions. We find that the effect (a) depends on the spouses' satisfaction with the division, (b) is partly moderated by relationship satisfaction, and (c) varies by parity. The division of household labor, however, seems of less importance for the realization of childbearing intentions.


Paracellular glucose transport plays a minor role in the unanesthetized dog.
Traditionally, intestinal glucose absorption was thought to occur through active, carrier-mediated transport. However, proponents of paracellular transport have argued that previous experiments neglected effects of solvent drag coming from high local concentrations of glucose at the brush-border membrane. The purpose of this study was to evaluate glucose absorption in the awake dog under conditions that would maximize any contribution of paracellular transport. Jejunal Thiry-Vella loops were constructed in six female mongrel dogs. After surgical recovery, isotonic buffers containing L-glucose as the probe for paracellular permeability were given over 2-h periods by constant infusion pump. At physiological concentrations of D-glucose (1-50 mM), the fractional absorption of L-glucose was only 4-7% of total glucose absorption. Infusion of supraphysiological concentrations (150 mM) of D-glucose, D-maltose, or D-mannitol yielded low-fractional absorptions of L-glucose (2-5%), so too did complex or nonabsorbable carbohydrates. In all experiments, there was significant fractional water absorption (5-19%), a prerequisite for solvent drag. Therefore, with even up to high concentrations of luminal carbohydrates in the presence of significant water absorption, the relative contribution of paracellular glucose absorption remained low.


Melatonin and adolescent idiopathic scoliosis.
Scoliosis seen in the chicken after pinealectomy resembles adolescent idiopathic scoliosis in man. It has been suggested that in both species, deficiency of the pineal hormone, melatonin, is responsible for this phenomenon. In nine patients with adolescent idiopathic scoliosis and in ten age- and gender-matched controls, the circadian levels of serum melatonin and the excretion of urinary 6-hydroxy-melatonin-sulphate, the principal metabolite of melatonin, were determined. There were no statistically significant differences in the secretion of serum melatonin or the excretion of urinary 6-hydroxy-melatonin-sulphate between the patients and the control group. The hypothesis of melatonin deficiency as a causative factor in the aetiology of adolescent idiopathic scoliosis cannot be supported by our data.


Species correlations between cardiac isomyosins. A comparison of electrophoretic and immunological properties.
Structural relationships between cardiac isomyosins were analyzed in 10 species using native-gel electrophoresis and radioimmunoassay. In the rat and rabbit, three types of ventricular isomyosin, V1, V2, and V3, were identified by electrophoresis. Monoclonal antibodies specific for the heavy chains of either type V1 or type V3 isomyosin in the rat and rabbit were used for comparison of immunological relationships between atrial and ventricular myosins in other species. Normal guinea pig ventricular myosin reacted with both anti-V2 and anti-V3 antibodies, but only a single myosin band was detected in this species by electrophoresis. When thyrotoxic cardiac hypertrophy was induced in guinea pigs, there was a decrease in myosin reactivity with the anti-V3 antibody and an increase in anti-V1 reactivity. This change in immunological reactivity indicated a change in proportions of two cardiac isomyosins in the guinea pig ventricle even though no myosin heterogeneity was detected by electrophoresis. In six other species including Xenopus, chicken, dog, pig, beef, and human, only a single band of myosin was detected by electrophoresis, and each myosin reacted only with the anti-V3 antibody. In the mouse, three types of ventricular myosin were also detected by electrophoresis. However, unlike V1 isomyosin of the rat and rabbit, mouse V1 isomyosin reacted equally with both anti-V1 and anti-V3 antibodies. In conclusion, we have identified highly conserved epitopes in cardiac myosin, which were found to specifically occur on either the high Ca2+-ATPase type V1 isomyosin or the lower ATPase type V3 ventricular isomyosin in most of the species examined.


Lomerizine attenuates LPS-induced acute lung injury by inhibiting the macrophage activation through reducing Ca2+ influx.
Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are life-threatening lung diseases with high mortality rates, predominantly attributable to acute and severe pulmonary inflammation. Lomerizine (LMZ) is a calcium channel blocker previously used in preventing and treating migraine. Here, we found that LMZ inhibited inflammatory responses and lung pathological injury by reducing pulmonary edema, neutrophil infiltration and pro-inflammatory cytokine production in lipopolysaccharide (LPS)-induced ALI mice. In vitro experiments, upon treating with LMZ, the expression of interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α was attenuated in macrophages. The phosphorylation of p38 MAPK, ERK1/2, JNK, and NF-κB p65 was inhibited after LMZ treatment. Furthermore, LPS-induced Ca2+ influx was reduced by treating with LMZ, which correlated with inhibition of pro-inflammatory cytokine production. And L-type Ca2+ channel agonist Bay K8644 (BK) could restore cytokine generation. In conclusion, our study demonstrated that LMZ alleviates LPS-induced ALI and is a potential agent for treating ALI/ARDS.


Effects of alcohol consumption on eight circulating markers of liver fibrosis.
A number of circulating breakdown products of collagen or other components of extracellular matrix, matrix degrading metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) have been proposed as markers of hepatic fibrosis. However, the published results lack consistency. Since many of the patients with fibrosis studied were alcoholics, the question was raised whether recent alcohol consumption may affect the results obtained. Using sandwich-type assays of radioimmunoassay technology with corresponding antibodies, we studied eight markers of liver fibrosis: laminin, tenascin, undulin, TIMP-1, collagen VI, procollagen type III (PIIINP), hyaluronic acid (HA) and MMP-2. A group of 10 alcoholics was studied after significant alcohol consumption and following 2 weeks of abstinence, verified with repeated breath alcohol measurement. Laminin was significantly reduced at 1 week (22%) and at 2 weeks (30%). Similarly, tenascin and undulin were also significantly decreased. By contrast, TIMP-1, collagen VI, PIIINP, HA and MMP-2 did not significantly change. The mode of action of alcohol on these tests is unknown. These differences must be considered when using those measurements to assess liver fibrosis.


Multidrug-Resistant Tuberculous Mediastinal Lymphadenitis, with an Esophagomediastinal Fistula, Mimicking an Esophageal Submucosal Tumor.
Mediastinal tuberculous lymphadenitis rarely mimics esophageal submucosal tumor, particularly in the case of multidrug-resistant tuberculosis (MDR-TB). Herein, we report the case of a 61-year-old woman who visited a local hospital complaining of odynophagia. An initial esophagogastroduodenoscopy revealed an esophageal submucosal tumor, and subsequent chest computed tomography showed subcarinal lymphadenopathy with an esophagomediastinal fistula. The patient was then referred to Samsung Medical Center, and a second esophagogastroduodenoscopy showed deep central ulceration, as well as a suspicious fistula in the esophageal submucosal tumor-like lesion. A biopsy examination of the ulcerative lesion confirmed focal inflammation only. Next, an endobronchial, ultrasound-guided lymph node biopsy was performed, and TB was confirmed. The patient initially began a course of isoniazid, rifampicin, ethambutol, and pyrazinamide. However, after a drug sensitivity test, she was diagnosed with MDR-TB, and second-line anti-TB medications were prescribed. She recovered well subsequently.


Taste discriminating capability to different bitter compounds by the larval styloconic sensilla in the insect herbivore Papilio hospiton (Géné).
Herbivorous animals may benefit from the capability to discriminate the taste of bitter compounds since plants produce noxious compounds, some of which toxic, while others are only unpalatable. Our goal was to investigate the contribution of the peripheral taste system in the discrimination of different bitter compounds by an herbivorous insect using the larvae of Papilio hospiton Géné as the experimental model, showing a narrow choice range of host plants. The spike activity from the lateral and medial styloconic sensilla, housing two and one bitter-sensitive gustatory receptor neurons (GRNs), respectively, was recorded following stimulation with nicotine, caffeine, salicin and quercitrin and the time course of the discharges was analyzed. Nicotine and caffeine activated all three bitter-sensitive GRNs, while salicin and quercitrin affected only two of them. In feeding behavior bioassays, intact larvae ate glass-fiber disks moistened with salicin and quercitrin, but rejected those with nicotine and caffeine, while lateral sensillum-ablated insects also ate the disks with the two latter compounds. The capability to discriminate bitter taste stimuli and the neural codes involved are discussed.


Identification of cholera hotspots in Zambia: A spatiotemporal analysis of cholera data from 2008 to 2017.
The global burden of cholera is increasing, with the majority (60%) of the cases occurring in sub-Saharan Africa. In Zambia, widespread cholera outbreaks have occurred since 1977, predominantly in the capital city of Lusaka. During both the 2016 and 2018 outbreaks, the Ministry of Health implemented cholera vaccination in addition to other preventative and control measures, to stop the spread and control the outbreak. Given the limitations in vaccine availability and the logistical support required for vaccination, oral cholera vaccine (OCV) is now recommended for use in the high risk areas ("hotspots") for cholera. Hence, the aim of this study was to identify areas with an increased risk of cholera in Zambia. Retrospective cholera case data from 2008 to 2017 was obtained from the Ministry of Health, Department of Public Health and Disease Surveillance. The Zambian Central Statistical Office provided district-level population data, socioeconomic and water, sanitation and hygiene (WaSH) indicators. To identify districts at high risk, we performed a discrete Poisson-based space-time scan statistic to account for variations in cholera risk across both space and time over a 10-year study period. A zero-inflated negative binomial regression model was employed to identify the district level risk factors for cholera. The risk map was generated by classifying the relative risk of cholera in each district, as obtained from the space-scan test statistic. In total, 34,950 cases of cholera were reported in Zambia between 2008 and 2017. Cholera cases varied spatially by year. During the study period, Lusaka District had the highest burden of cholera, with 29,080 reported cases. The space-time scan statistic identified 16 districts to be at a significantly higher risk of having cholera. The relative risk of having cholera in these districts was significantly higher and ranged from 1.25 to 78.87 times higher when compared to elsewhere in the country. Proximity to waterbodies was the only factor associated with the increased risk for cholera (P<0.05). This study provides a basis for the cholera elimination program in Zambia. Outside Lusaka, the majority of high risk districts identified were near the border with the DRC, Tanzania, Mozambique, and Zimbabwe. This suggests that cholera in Zambia may be linked to movement of people from neighboring areas of cholera endemicity. A collaborative intervention program implemented in concert with neighboring countries could be an effective strategy for elimination of cholera in Zambia, while also reducing rates at a regional level.


An analysis of immediate postmastectomy breast reconstruction frequency using the surveillance, epidemiology, and end results database.
Mastectomy is used to treat one third of the nearly 180,000 women diagnosed with breast cancer in the United States annually. In this study, we use population-level data from multiple years of the Surveillance, Epidemiology, End Results (SEER) database to further define patient, tumor, and geographic characteristics associated with immediate and early-delayed breast reconstruction. Population level de-identified data for the years 1998 to 2002 were extracted from the National Cancer Institute's (NCI) SEER cancer database. All female patients who were treated with mastectomy for a diagnosis of ductal and/or lobular breast cancer (including Paget disease) were included. The primary end point of interest was odds of reconstruction. Multivariate analysis was performed to control for patient demographic and oncologic characteristics. A total of 52,249 patients met the inclusion criteria. Reconstruction was performed in 8,446 patients (16.2%). Odds of reconstruction varied by region from 0.60 (Seattle) to 2.81 (Atlanta). African Americans were noted to have a significantly lower likelihood of reconstruction when compared with Caucasian patients (OR 0.60 versus 1.00). Patients living in nonmetropolitan regions were also significantly less likely to undergo reconstruction. Receipt of radiation therapy was also negatively correlated with likelihood of reconstruction. In this multicenter, multiyear analysis of factors associated with immediate or early-delayed reconstruction after mastectomy, we demonstrate that younger age, white race, metropolitan locale, and lower stage disease were all independently associated with higher likelihood of reconstruction. This information provides insight into breast reconstruction utilization and will help guide future studies to understand how these factors affect patient and physician decision-making.


Rupture of intracranial aneurysms during treatment with Guglielmi detachable coils: incidence, outcome, and risk factors.
The aim of this study was to assess the incidence and outcome of procedure-related rupture of intracranial aneurysms in patients treated with Guglielmi detachable coils (GDCs) and to identify risk factors for this complication. Procedure-related rupture occurred in seven of 264 treated aneurysms in 239 consecutive patients. Aneurysm size, history of previous subarachnoid hemorrhage (SAH) caused by the treated aneurysm, timing of treatment after SAH, and the use of a temporary occlusion balloon in the seven procedures in which rupture occurred were compared with the remaining 257 procedures, and these findings were correlated with data from 13 studies in the literature, in which results of 2030 aneurysm treatments were reported. Procedure-related rupture of intracranial aneurysms during GDC treatment occurs in 2.5% of cases and is responsible for 1% of treatment-related deaths. Risk factors are as follows: small aneurysm size, previous SAH, and probably the use of a temporary occlusion balloon.


Knowledge-Guided Multi-Label Few-Shot Learning for General Image Recognition.
Recognizing multiple labels of an image is a practical yet challenging task, and remarkable progress has been achieved by searching for semantic regions and exploiting label dependencies. However, current works utilize RNN/LSTM to implicitly capture sequential region/label dependencies, which cannot fully explore mutual interactions among the semantic regions/labels and do not explicitly integrate label co-occurrences. In addition, these works require large amounts of training samples for each category, and they are unable to generalize to novel categories with limited samples. To address these issues, we propose a knowledge-guided graph routing (KGGR) framework, which unifies prior knowledge of statistical label correlations with deep neural networks. The framework exploits prior knowledge to guide adaptive information propagation among different categories to facilitate multi-label analysis and reduce the dependency of training samples. Specifically, it first builds a structured knowledge graph to correlate different labels based on statistical label co-occurrence. Then, it introduces the label semantics to guide learning semantic-specific features to initialize the graph, and it exploits a graph propagation network to explore graph node interactions, enabling learning contextualized image feature representations. Moreover, we initialize each graph node with the classifier weights for the corresponding label and apply another propagation network to transfer node messages through the graph. In this way, it can facilitate exploiting the information of correlated labels to help train better classifiers, especially for labels with limited training samples. We conduct extensive experiments on the traditional multi-label image recognition (MLR) and multi-label few-shot learning (ML-FSL) tasks and show that our KGGR framework outperforms the current state-of-the-art methods by sizable margins on the public benchmarks.


Sub-inhibitory concentrations of different pharmaceutical products affect the meta-transcriptome of river biofilm communities cultivated in rotating annular reactors.
Surface waters worldwide are contaminated by pharmaceutical products that are released into the environment from wastewater treatment plants. Here, we hypothesize that pharmaceutical products have effects on organisms as well as genes related to nutrient cycling in complex microbial communities. To test this hypothesis, biofilms were grown in reactors and subjected low concentrations of three antibiotics [erythromycin, ER, sulfamethoxazole, SL and sulfamethazine, SN) and a lipid regulator (gemfibrozil, GM). Total community RNA was extracted and sequenced together with PCR amplicons of the 16S rRNA gene using 454 pyrosequencing. Exposure to pharmaceutical products resulted in very little change in bacterial community composition at the phylum level based on 16S rRNA gene amplicons, even though some genera were significantly affected. In contrast, large shifts were observed in the active community composition based on taxonomic affiliations of mRNA sequences. Consequently, expression of gene categories related to N, P and C cycling were strongly affected by the presence of pharmaceutical products, with each treatment having specific effects. These results indicate that low pharmaceutical product concentrations rapidly provoke a variety of functional shifts in river bacterial communities. In the longer term these shifts in gene expression and microbial activity could lead to a disruption of important ecosystem processes like nutrient cycling.


[Treatment of anterior glenohumeral instability: personal experience with an arthroscopic stabilization technique, its indications and results].
Arthroscopic treatment of anterior post-traumatic instability of the glenohumeral joint is a recent surgical procedure. The aim of this study was to evaluate the outcomes of the method and define criteria on which indications for this treatment are based. Fifty-six active, young patients less than 30 years of age were distributed into three groups according to the method of treatment. Twenty patients were treated conservatively (group A), 12 patients had open surgery (B) and 23 patients underwent arthroscopic stabilization of the glenohumeral joint (C). The average follow-up periods in groups A, B and C were 26, 38 and 18 months, respectively. The treatment of all patients in groups B and C was preceded by physical therapy lasting at least 3 months. Contraindications to arthroscopic treatment included Hill-Sachs bony defects, instability involving a fracture of the glenoid cavity, conditions after previous surgery, posterior or multidirectional instability and anterior instability due to a full rupture of the rotator cuff. Conservative treatment consisted of early closed reduction, 3 to 4 weeks of immobilization and subsequent physical therapy. Open procedures for Bankart lesions were carried out according to either Cave and Rowe or Bristow, with post-operative immobilization for 4 weeks. Arthroscopic stabilization was performed by the Wolf method. After the size of the defect had been identified, the glenoid rim was scratched to bleed, the detached labrum, including the inferior glenohumeral ligament, was mobilized and, after drilling holes in the glenoid rim, the capsulolabral complex was fixed by means of a Mitek GII anchor with a 1-0 PDS fibre. Three anchors were inserted as a rule. In each group, the number of recurrent dislocations was recorded. In the patients undergoing surgery, the loss of passive external rotation in 90 degrees abduction was assessed and the outcome was evaluated according to the Rowe rating system. Recurrent dislocations were experienced by 13 patients (65%) in group A and two patients (8.7%) in group C; no recurrent dislocation occurred in group B patients. The average loss of external rotation in 90 degrees abduction was 11.3 degrees and 6.7 degrees in groups B and C, respectively. The Rowe scores showed an excellent outcome in 80%, good in 8%, satisfactory in 2% and poor outcomes in 10% of the group B patients; in group C 78% had excellent, 7% had good, 8% had satisfactory, and 7% had poor outcomes. The number of recurrent dislocations (8.7%) in our patients treated by arthroscopy was in agreement with the literature data (1 to 40%); both figures refer to recurrent dislocations in subjects involved in body-contact sports. In our patients treated by arthroscopy, the average post-operative loss of external rotation in 90 degrees abduction was lower than in the open surgery group. There were no differences in the Rowe scores between the two surgically treated groups. The patients treated conservatively showed a high number of recurrent dislocations (65%), thus confirming reports by other authors on the failure of this method. The arthroscopic treatment of anterior post-traumatic glenohumeral instability, using the Wolf method, resulted in a reduction of recurrent dislocations, supposing indication criteria were observed. Its outcomes were comparable with the results of conventional open surgery.


Cervicogenic headache arising from hidden metastasis to cervical lymph node adjacent to the superficial cervical plexus -A case report-.
The differential diagnosis of headache is often difficult because the symptom of headache is overlapping. Superficial cervical plexus block is useful in diagnosis and treatment of headache. Headache arising from the neck and radiating to the frontotemporal regions and possibly to the supraorbital region has been defined as cervicogenic headache. A positive response to anesthetic blocks is one of the diagnostic criteria of cervicogenic headache. We experienced a case of headache arising from direct lymph node metastasis of hepatocellular carcinoma adjacent to the superficial cervical plexus during treatment of cervicogenic headache under ultrasonographic guidance. Especially in patients with medical history of cancer, practitioners should consider the possibility of metastasis to cervical lymph nodes and using ultrasonography to evaluate the cervical area prior to the practice.


Outcomes of systemic to pulmonary artery shunts in patients weighing less than 3 kg: analysis of shunt type, size, and surgical approach.
To evaluate outcomes of systemic to pulmonary artery shunts (SPS) in patients weighing less than 3 kg with regard to shunt type, shunt size, and surgical approach. Patients weighing less than 3 kg who underwent modified Blalock-Taussig or central shunts with polytetrafluoroethylene grafts at our institution from January 1, 2000, to May 31, 2011, were reviewed. Patients who had undergone other major concomitant procedures were excluded from the analysis. Primary outcomes included mortality (discharge mortality and mortality before next planned palliative procedure or definitive repair), cardiac arrest and/or extracorporeal membrane oxygenation (ECMO), and shunt reintervention. In this cohort of 80 patients, discharge survival was 96% (77/80). Postoperative cardiac arrest or ECMO occurred in 6/80 (7.5%), and shunt reintervention was required in 14/80 (17%). On univariate analysis, shunt reintervention was more common in patients with 3-mm shunts (11/30, 37%) compared with 3.5-mm (2/36, 6%) or 4-mm shunts (1/14, 7%) (P < .003). There were no statistically significant associations between shunt type, shunt size, or surgical approach and cardiac arrest/ECMO or mortality. Multiple logistic regression demonstrated that a shunt size of 3 mm (P = .019) and extracardiac anomaly (P = .047) were associated with shunt reintervention, whereas no variable was associated with cardiac arrest/ECMO or mortality. In this high-risk group of neonates weighing less than 3 kg at the time of SPS, survival to discharge and the next planned surgical procedure was high. Outcomes were good with the 3.5- and 4-mm shunts; however, shunt reintervention was common with 3-mm shunts.


Antibacterial immune functions of subadults and adults in a semelparous spider.
Although capacity to mount an efficient immune response plays a critical role in individuals' survival, its dynamics across ontogenetic stages is still largely unexplored. Life stage-dependent variation in the encountered diversity and prevalence of parasites were proposed to contribute to stage-dependent changes in immunity, but differences in life history objectives between developmental stages may also lead to stage-specific changes in efficiency of given immune mechanisms. The reason for this is that juveniles and subadults are unable to reproduce, therefore they invest resources mainly into survival, while adults have to partition their resources between survival and reproduction. The general trade-off between somatic maintenance and reproductive effort is expected to impair immune function. Especially so in semelparous organisms that only reproduce once throughout their lifetime, hence they do not face the trade-off between current and future reproduction. We hypothesised that in a semelparous species individuals would be characterised by decreased investment into somatic maintenance after maturation, in order to maximise their reproductive output. Accordingly, we predicted that (1) elements of somatic maintenance, such as immunity, should be relatively weaker in adults in comparison to subadults, and (2) increased reproductive investment in adults should be associated with lower immune efficiency. We quantified two markers of immunity in subadult and adult individuals of the semelparous wolf spider Pardosa agrestis (Westring, 1861), namely bacterial growth inhibition power and bacterial cell wall lytic activity. We found that subadults showed significantly higher cell wall lytic activity than adults, but the two life stages did not differ in their capacity to inhibit bacterial growth. Also, we found weaker immune measures in mated females compared to virgins. Furthermore, in mated females bacterial growth inhibition power was negatively associated with fecundity.


A Novel WT1 Mutation Identified in a 46,XX Testicular/Ovotesticular DSD Patient Results in the Retention of Intron 9.
The 46,XX testicular DSD (disorder/difference of sexual development) and 46,XX ovotesticular DSD (46,XX TDSD and 46,XX OTDSD) phenotypes are caused by genetic rearrangements or point mutations resulting in imbalance between components of the two antagonistic, pro-testicular and pro-ovarian pathways; however, the genetic causes of 46,XX TDSD/OTDSD are not fully understood, and molecular diagnosis for many patients with the conditions is unavailable. Only recently few mutations in the WT1 (WT1 transcription factor; 11p13) gene were described in a group of 46,XX TDSD and 46,XX OTDSD individuals. The WT1 protein contains a DNA/RNA binding domain consisting of four zinc fingers (ZnF) and a three-amino acid (KTS) motif that is present or absent, as a result of alternative splicing, between ZnF3 and ZnF4 (±KTS isoforms). Here, we present a patient with 46,XX TDSD/OTDSD in whom whole exome sequencing revealed a heterozygous de novo WT1 c.1437A>G mutation within an alternative donor splice site which is used for -KTS WT1 isoform formation. So far, no mutation in this splice site has been identified in any patient group. We demonstrated that the mutation results in the retention of intron 9 in the mature mRNA of the 46,XX TDSD/OTDSD patient. In cases when the erroneous mRNA is translated, exclusively the expression of a truncated WT1 +KTS protein lacking ZnF4 and no -KTS protein occurs from the mutated allele of the patient. We discuss potential mechanisms and pathways which can be disturbed upon two conditions: Absence of Zn4F and altered +KTS/-KTS ratio.


Alcohol, allopregnanolone and aggression in mice.
Aggressive behavior of certain individual animals can be greatly increased when under the influence of low doses of alcohol. One of alcohol's neurochemical actions that may be relevant to alcohol-heightened aggression (AHA) is its positive modulation of the GABA(A) receptor complex. The objective of this study was to investigate whether alcohol interacts with an endogenous modulator of the GABA(A) receptor complex, the neurosteroid allopregnanolone, in stimulating/heightening aggressive behavior. The first experiment was designed to test the hypothesis that neurosteroid modulators of the GABA(A) receptor complex will increase aggression and to compare these effects with alcohol. Male CFW mice were injected with allopregnanolone, alphaxalone (3-30 mg/kg, i.p.), or alcohol (1.0 g/kg, p.o.) 15 min prior to a 5-min confrontation with an intruder. Moderate doses of alcohol and the neurosteroids increased aggression by ca. 50% above baseline; impaired locomotion was seen only at the highest doses. A second experiment compared AHA and ANA (i.e. alcohol-non-heightened aggression) mice by giving allopregnanolone (1-10 mg/kg) with a simultaneous oral injection of alcohol (0.6 or 1.0 g/kg) or water. When administered with water and the 0.6 g/kg dose of alcohol, allopregnanolone increased the aggression of AHA and ANA mice. Administration of the 1.0 g/kg dose of alcohol in ANA mice prevented allopregnanolone-heightened aggression. In AHA mice, addition of allopregnanolone to 1.0 g/kg alcohol dose-dependently reduced alcohol-heightened aggression, suggesting potentiation of alcohol's suppressive effects on aggression. The neuroactive steroid allopregnanolone appears to play an important role in alcohol-heightened aggression. Moreover, the upward shift of the aggression-heightening effects of alcohol and the downward shift at the maximally effective alcohol dose by allopregnanolone point to a shared mechanism for both positive modulators of the GABA(A) receptor complex.


Cyclosporine treatment of RPE allografts in the rabbit subretinal space.
To determine the effects of systemic cyclosporine A (CsA) on the survival of retinal pigment epithelial (RPE) allografts in the subretinal space in an animal model using atraumatic transplantation surgery. Following pars plana vitrectomy, an RPE cell suspension from brown rabbits was injected with a glass micropipette into the subretinal space of 39 albino rabbits. For immunosuppression, 22 rabbits were given an injection of CsA, 20 mg daily intramuscularly, 17 rabbits with RPE grafts were controls. The grafts were monitored by biomicroscopy, color fundus photography, and fluorescein angiography. Rabbits were sacrificed at 1, 3 and 6 months, respectively, and the eyes processed for light and electron microscopy including immunohistochemistry. After three months, the transplanted RPE cells, in both the CsA group and the controls, formed a monolayer in the subretinal space. Although a few macrophages were encountered, there was no massive cellular infiltration and the photoreceptor layer was well preserved. After six months, however, there was a disruption of grafted RPE cells in both groups, characterized by dispersion of melanin pigment in the subretinal space, and invasion of macrophages with focal photoreceptor damage but no infiltration of lymphocytes in the retina or choroid. No significant differences between the CsA treated and the control eyes were discernible. Although the subretinal space has been considered an immunologically privileged site, we found that the survival of RPE allografts was limited. CsA did not prevent RPE allograft destruction in the subretinal space. The transplant seems to be disrupted either by immunological mechanisms that are not inhibited by CsA, or by nonimmunologic events.


Children with hemodynamically significant congenital heart disease can be identified through population-based registers.
Epidemiological research is facilitated in Sweden by a history of national health care registers, making large unselected national cohort studies possible. However, for complex clinical populations, such as children with congenital heart disease (CHD), register-based studies are challenged by registration limitations. For example, the diagnostic code system International Classification of Diseases, 10th version (ICD-10) does not indicate the clinical significance of abnormalities, therefore may be of limited use if used as the sole parameter in epidemiological research. Palivizumab is indicated as a prophylactic treatment against respiratory syncytial virus infections in children with hemodynamically significant CHD. The aim of the study reported here was to develop and validate an algorithm to identify children with hemodynamically significant CHD according to recommendations for palivizumab prophylaxis in register-based research. By using a strategy of combining criteria for age at diagnosis, diagnostic codes, surgical procedure codes, and dispensing records, we created an algorithm to define the specific cases with hemodynamically significant CHD in which palivizumab could be advocated according to recommendations. The algorithm identified 928 children with hemodynamically significant CHD in the Swedish birth cohort born July 1, 2005 to December 31, 2010. A sensitivity (95% confidence interval) of 80% (70-88) for the algorithm was found by analyzing 121 children identified through local hospital data who were treated with palivizumab within a defined region and study period. The positive predictive value was estimated by medical record review in a random sample of 34 cases identified by the algorithm. In 79% (62-91) of these cases, the children were regarded as having hemodynamically significant CHD according to the recommendations for treatment with palivizumab. It was possible to identify a subgroup of children with hemodynamically significant CHD using an epidemiological approach and an algorithm with high validity. Our results will enable well-powered national cohort studies of individuals with complex clinical conditions such as hemodynamically significant CHD.


Diagnostic Pathways to Alzheimer Disease: Costs Incurred in a Medicare Population.
Despite its implications on the personal and policy level, little is currently known about the specific diagnostic pathways that patients with cognitive impairment (CI) pass through before being diagnosed with Alzheimer disease (AD). Four major diagnostic pathways were identified in the Medicare claims records for 2001 to 2006: AD as initial diagnosis, cognitive disturbance followed by AD; dementia with suspected etiologies followed by AD; dementia without known cause followed by AD; and 1 triple pathway, cognitive disturbance followed by dementia without known cause followed by AD. For all of these pathways, previously low medical costs peaked during patients' month of initial diagnosis and then declined to a level substantially higher than before. The 3 CI pathways that transition to AD included another peak in costs when a secondary AD diagnosis occurred. Each time, inpatient and skilled nursing facility services were major cost contributors. The primary diagnoses on Medicare claims for the AD event were usually comorbidities rather than CI. A linear regression model adjusting for demographic factors, selected comorbidities, and overall frailty found that the transitional CI diagnoses were significant independent cost determinants. They increased Medicare expenditures by an estimated $4600 to $14,200 relative to patients whose initial CI diagnosis was AD.


Molecular, immunological, and structural characterization of Phl p 6, a major allergen and P-particle-associated protein from Timothy grass (Phleum pratense) pollen.
Due to the wide distribution and heavy pollen production of grasses, approximately 50% of allergic patients are sensitized against grass pollen allergens. cDNAs coding for two isoforms and four fragments of a major timothy grass (Phleum pratense) pollen allergen, Phl p 6, were isolated by IgE immunoscreening from a pollen expression cDNA library. Recombinant Phl p 6 (rPhl p 6), an acidic protein of 11.8 kDa, was purified to homogeneity as assessed by mass spectrometry and exhibited almost exclusive alpha-helical secondary structure as determined by circular dichroism spectroscopy. Phl p 6 reacted with serum IgE from 75% of grass pollen-allergic patients (n = 171). IgE binding experiments with rPhl p 6 fragments indicated that the N terminus of the allergen is required for IgE recognition. Purified rPhl p 6 elicited dose-dependent basophil histamine release and immediate type skin reactions in patients allergic to grass pollen. A rabbit antiserum raised against purified rPhl p 6 identified it as a pollen-specific protein that, by immunogold electron microscopy, was localized on the polysaccharide-containing wall-precursor bodies (P-particles). The association of Phl p 6 with P-particles may facilitate its intrusion into the deeper airways and thus be responsible for the high prevalence of IgE recognition of Phl p 6. Recombinant native-like Phl p 6 can be used for in vitro as well as in vivo diagnoses of grass pollen allergy, whereas N-terminal deletion mutants with reduced IgE binding capacity may represent candidates for immunotherapy of grass pollen allergy with a low risk of anaphylactic side effects.


Quality assurance program for peripheral blood mononuclear cell cryopreservation.
Seven Brazilian sites participating in the Pediatric AIDS Clinical Trials Group international cryopreservation quality assurance pilot program cryopreserved and shipped peripheral blood mononuclear cells (PBMC) to a central U.S. laboratory for analysis. Cell viability and recovery significantly increased over time. A wet-laboratory training session conducted at the central laboratory significantly improved the quality of the cryopreserved PBMC.


Incorporation of endothelial progenitor cells into mosaic pseudoislets.
Pancreatic islet transplantation is limited by extensive apoptosis and suboptimal function of the implanted islets in the longer term. Endothelial progenitor cells (EPC) may be ideal for enhancing both the survival and function of transplanted islets. Here, we describe for the first time the in vitro formation of rat mosaic pseudoislets comprised of pancreatic β-cells with interspersed vasculogenic EPC. Bone marrow-derived EPC displayed a similar phenotype to non-adherent EPC, recently described in the human and mouse. Mosaic pseudoislet formation was enhanced by the use of an embryoid body forming medium (BPEL) and a spin protocol. Mosaic pseudoislets maintained function in vitro and may represent an enhanced cell therapy delivery approach to enhance the survival and revascularisation of transplanted islets.


Fluorescent in situ hybridization assessment of chromosome 8 copy number in breast cancer.
Conventional cytogenetics of breast and other solid tumors has been hampered by a number of factors. An analysis of breast tumor tissues was therefore undertaken using fluorescent in situ hybridization (FISH). A total of 34 specimens were analyzed using a chromosome 8-specific alpha-satellite probe. Various approaches were tested and compared. Among 30 informative samples, 11 infiltrating ductal carcinomas, not otherwise specified (NOS), 5 ductal carcinomas in situ, 5 lobular carcinomas, 3 papillary carcinomas, and 6 benign lesions were studied. Of the 11 cases of infiltrating ductal carcinomas (NOS) analyzed, four cases showed 3 signals, one case showed 4 signals, and the rest showed 2 signals. Of the 5 cases of ductal carcinoma in situ samples, 1 showed 3 signals and the other 4 cases showed 2 signals. All cases of lobular carcinomas, papillary carcinomas, and benign lesions showed 2 signals. We inferred from these data that 36% of the infiltrating ductal carcinomas (NOS) were trisomic and 9% were tetrasomic, whereas 20% of the ductal carcinomas in situ were trisomic. All samples from lobular carcinomas, papillary carcinomas, and the benign lesions were disomic. From our preliminary data, it can further be concluded that a subset of breast cancer is characterized by chromosome 8 trisomy. These data are consistent with an ever-increasing database on the association of chromosomal 8 trisomy with other cancers such as leukemia, lymphoma, prostate cancer, ovarian carcinoma, salivary gland tumor, malignant melanoma, desmoid tumors, and recently gestational trophoblastic disease. It is also noted that the ability to analyze formalin-fixed, paraffin-embedded archival material will enable a more comprehensive cytogenetic study of breast cancer than is currently available.


Mechanism of the hydroxyl radical oxidation of methacryloyl peroxynitrate (MPAN) and its pathway toward secondary organic aerosol formation in the atmosphere.
Methacryloyl peroxynitrate (MPAN), the acyl peroxynitrate of methacrolein, has been suggested to be an important secondary organic aerosol (SOA) precursor from isoprene oxidation. Yet, the mechanism by which MPAN produces SOA through reaction with the hydroxyl radical (OH) is unclear. We systematically evaluate three proposed mechanisms in controlled chamber experiments and provide the first experimental support for the theoretically-predicted lactone formation pathway from the MPAN + OH reaction, producing hydroxymethyl-methyl-α-lactone (HMML). The decomposition of the MPAN-OH adduct yields HMML + NO3 (∼75%) and hydroxyacetone + CO + NO3 (∼25%), out-competing its reaction with atmospheric oxygen. The production of other proposed SOA precursors, e.g., methacrylic acid epoxide (MAE), from MPAN and methacrolein are negligible (<2%). Furthermore, we show that the beta-alkenyl moiety of MPAN is critical for lactone formation. Alkyl radicals formed cold via H-abstraction by OH do not decompose to HMML, even if they are structurally identical to the MPAN-OH adduct. The SOA formation from HMML, from polyaddition of the lactone to organic compounds at the particle interface or in the condensed phase, is close to unity under dry conditions. However, the SOA yield is sensitive to particle liquid water and solvated ions. In hydrated inorganic particles, HMML reacts primarily with H2O to produce the monomeric 2-methylglyceric acid (2MGA) or with aqueous sulfate and nitrate to produce the associated organosulfate and organonitrate, respectively. 2MGA, a tracer for isoprene SOA, is semivolatile and its accommodation in aerosol water decreases with decreasing pH. Conditions that enhance the production of neutral 2MGA suppress SOA mass from the HMML channel. Considering the liquid water content and pH ranges of ambient particles, 2MGA will exist largely as a gaseous compound in some parts of the atmosphere.


Timing jitter and phase noiseof mode-locked fiber lasers.
The noise properties of mode-locked fiber lasers differ in various respects from those of bulk lasers. The reasons for this are both quantitative and qualitative differences concerning the pulse formation. The underlying theoretical aspects are discussed in detail. It is found that the achievable noise level and the limiting effects depend strongly on the type of fiber laser. Depending on the pulse formation mechanism, noise levels may be much higher than predicted by simplified models.


N-Ethyl-N-nitrosourea induces mammary cancers in the pituitary-isografted mouse which are histologically and genotypically distinct from those induced by N-methyl-N-nitrosourea.
N-Ethyl-N-nitrosourea (ENU) and N-methyl-N-nitrosourea (MNU) are alkylating agents which respectively ethylate or methylate nucleophilic centers in the cell such as DNA. In vitro studies with naked DNA and bacterial mutagenesis assays suggest that these two compounds induce different spectra of genetic lesions. In addition, the ethyl-DNA adducts induced by ENU persist longer than the methyl-DNA adducts induced by MNU. Since MNU is a known mammary carcinogen in the pituitary-isografted mouse, these data suggest that ENU may be an even more potent carcinogen than MNU. The purpose of this study was to determine whether ENU was a mammary carcinogen in the pituitary-isografted mouse and if so, to compare the genotype and phenotype of ENU-induced mammary tumors with those induced by MNU. Fifteen adult female virgin BALB/c mice were isografted with two pituitaries and subsequently treated with a single intravenous injection of ENU (50 micrograms/g body weight). Mammary adenocarcinomas arose in all of the survivors (n=12) with a median latency of 27 weeks and a mean frequency of 1.4 cancers per mouse. When tumor DNA was analyzed for mutations in the 12th and 61st codons of c-Ki-ras or c-Ha-ras protooncogenes, only wild type sequences were found. This is in contrast to MNU which causes a G to A transition mutation in the 12th codon of the c-Ha-ras proto-oncogene in about one of five mammary cancers induced in pituitary-isografted mice. Furthermore, the ENU-induced tumors were solid viable papillary adenocarcinomas, whereas MNU induced tumors are highly necrotic adenocarcinomas with squamous metaplasia. These results demonstrate that, in the pituitary-isografted mouse, ENU is as potent a mammary carcinogen as MNU and suggest that oncogenes other than c-Ki-ras or c-Ha-ras may be involved in ENU-induced mammary cancers.


Effects of different cereal grains in diets for laying hens on production parameters and liver fat content.
Two experiments were conducted with White Leghorn laying hens to study the effects of different cereal grains on production criteria and liver fat content. The results of Experiment 1, in which pullets 21 weeks of age were used for a period of 22 weeks indicated that Gaines wheat or triticale (Trailblazer) were equal to corn in supporting egg production, egg weight and body weight, with comparable feed consumption. Henry wheat(a hard red winter class) was slightly, yet significantly (P less than 0.05) inferior to corn for the above criteria. No significant differences were observed among four treatments in wet liver weight and liver fat content. Hens fed the corn diet had significantly (P less than 0.05) lower carcass fat followed by the hens fed triticale in comparison with those fed Gaines or Henyry wheat. Mortality was very low and not related to dietary treatments. Neither dietary fat nor energy content was related to fat content of liver and carcass of the hens. Body weight and liver fat content were not closely related to each other. Wet liver weight was the only significantly (P less than 0.05) related factor to liver fat content. In the second experiment, in which hens 33 weeks of age were used for an experimental period of 20 weeks, opaque-2 corn was slightly superior to normal corn and triticale was comparable to normal corn in supporting egg production and egg weight. Supplementation of the diets containing the two corns and triticale with lysine failed to improve egg production and egg weight. Hens fed the diets containing either normal corn or opaque-2 corn as the only grain in the diet had significantly (P less than 0.05) higher liver fat content in comparison with hens fed the diet containing triticale as the only grain. Mortality, however, was much higher among hens fed triticale-containing diets in comparison with groups fed corn-containing diets in spite of the fact that they had significantly lower liver fat content. Regardless of dietary treatments or grains used, the hens that died were diagnosed to have fatty liver hemorrhagic syndrome. Dietary fat content was postively and significantly (P less than 0.05) related to liver fat content. Dietary energy or body weight was not closely related to liver content. Liver fat content and mortality were negatively related to each other. The higher for content did not adversely affect laying performance.


Ixekizumab improves spinal pain, function, fatigue, stiffness, and sleep in radiographic axial Spondyloarthritis: COAST-V/W 52-week results.
This analysis assessed improvements in patients with radiographic axial spondyloarthritis (r-axSpA) treated with ixekizumab in the Assessment of Spondyloarthritis International Society (ASAS) treatment response domains and additional patient-reported outcomes at 1 year of treatment. COAST-V and COAST-W were 52-week, phase 3, randomized controlled trials evaluating the efficacy and safety of ixekizumab in biologic disease-modifying antirheumatic drug (bDMARD)-naïve and tumor necrosis factor inhibitor (TNFi)-experienced patients with radiographic spondyloarthritis, respectively. Patients were treated with 80-mg ixekizumab either every 2 weeks or every 4 weeks. Patient-reported outcomes included Patient Global Disease Activity, Spinal Pain, stiffness as measured by Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) Questions 5 and 6, function as measured by the Bath Ankylosing Spondylitis Functional Index, fatigue as measured by the Fatigue Numeric Rating Scale and BASDAI question 1, Spinal Pain at Night, and sleep quality as measured by the Jenkins Sleep Evaluation Questionnaire. Mixed-effects models for repeated measures were used to analyze changes from baseline in patient-reported outcomes from weeks 1 to 16, and descriptive statistics were reported from weeks 20 to 52. Analysis of covariance with Scheffé's method was used for the ASAS response association analyses. This study assessed 341 bDMARD-naïve and 316 TNFi-experienced patients in the placebo-controlled blinded treatment dosing period (weeks 1-16) as well as 329 bDMARD-naïve and 281 TNFi-experienced patients in the dose double-blind extended treatment period (weeks 20-52). bDMARD-naïve or TNFi-experienced patients treated with ixekizumab every 2 weeks and every 4 weeks reported improvements in patient global disease activity, spinal pain, function, stiffness, fatigue, spinal pain at night, and sleep quality through week 52. Greater correlations with improvements in all response domains were seen when comparing ASAS40 responders to ASAS20 non-responders (p < 0.001), with up to 10.5-fold greater improvements observed in ASAS40 responses compared with ASAS20 non-responders. Function and fatigue demonstrated the highest values. Ixekizumab-treated bDMARD-naïve and TNFi-experienced patients with radiographic axial spondyloarthritis achieving ASAS40 reported sustained and consistent improvement in all ASAS response domains and other patient-reported outcomes though week 52, with spinal pain, function, and stiffness as major drivers of the response. NCT02696785 and NCT02696798 , March 2, 2016.


Computational modeling of chemo-electro-mechanical coupling: a novel implicit monolithic finite element approach.
Computational modeling of the human heart allows us to predict how chemical, electrical, and mechanical fields interact throughout a cardiac cycle. Pharmacological treatment of cardiac disease has advanced significantly over the past decades, yet it remains unclear how the local biochemistry of an individual heart cell translates into global cardiac function. Here, we propose a novel, unified strategy to simulate excitable biological systems across three biological scales. To discretize the governing chemical, electrical, and mechanical equations in space, we propose a monolithic finite element scheme. We apply a highly efficient and inherently modular global-local split, in which the deformation and the transmembrane potential are introduced globally as nodal degrees of freedom, whereas the chemical state variables are treated locally as internal variables. To ensure unconditional algorithmic stability, we apply an implicit backward Euler finite difference scheme to discretize the resulting system in time. To increase algorithmic robustness and guarantee optimal quadratic convergence, we suggest an incremental iterative Newton-Raphson scheme. The proposed algorithm allows us to simulate the interaction of chemical, electrical, and mechanical fields during a representative cardiac cycle on a patient-specific geometry, robust and stable, with calculation times on the order of 4 days on a standard desktop computer.


Identification and characterization of amino acid starvation-induced CD24 gene in MCF-7 human breast cancer cells.
Amino acid starvation is a pathophysiological condition that results in protein deprivation due to cancer cachexia. Using the method of differential display of reverse transcription PCR (DDRT-PCR), we isolated a cDNA fragment in MCF-7 human breast cancer cells in response to amino acid starvation, which was identical with human CD24 gene. Northern blot results showed that CD24 mRNA in MCF-7 cells was constitutively expressed and significantly upregulated upon amino acid starvation. This stimulation was time-dependent and the maximal response was at 24 h. The expression of the amino acid starvation-induced CD24 mRNA decreased when starved cells were returned to a medium supplemented with amino acids. This repressive response was also time-dependent. Amino acid starvation-induced CD24 mRNA expression in MCF-7 cells was completely blocked by actinomycin D, which suggested that the regulation of CD24 mRNA by amino acid availability occurred at transcriptional level. When amino acid-starved cells were refed with amino acids for 8 h, the expression of CD24 mRNA declined to the basal levels confirming that CD24 mRNA expression could be stimulated by amino acid starvation. Interestingly, CD24 mRNA was poorly detected in MCF-10 cells, a benign human breast epithelial cell line. In conclusion, CD24 mRNA expression in MCF-7 cells was upregulated upon amino acid starvation. This amino acid starvation-induced upregulation of CD24 mRNA occurred at transcriptional level. The regulation of CD24 mRNA in MCF-7 cells by amino acid availability may play an important role in the progression and metastasis of human breast cancer.


Prevalence of hypothyroidism in infertile women and evaluation of response of treatment for hypothyroidism on infertility.
Prevalence of hypothyroidism is 2-4% in women in the reproductive age group. Hypothyroidism can affect fertility due to anovulatory cycles, luteal phase defects, hyperprolactinemia, and sex hormone imbalance. To study the prevalence of clinical/sub-clinical hypothyroidism in infertile women and the response of treatment for hypothyroidism on infertility. A total of 394 infertile women visiting the infertility clinic for the first time were investigated for thyroid stimulating hormone (TSH) and prolactin (PRL). Infertile women with hypothyroidism alone or with associated hyperprolactinemia were given treatment for hypothyroidism with thyroxine 25-150 μg. Of 394 infertile women, 23.9% were hypothyroid (TSH > 4.2 μIU/ml). After treatment for hypothyroidism, 76.6% of infertile women conceived within 6 weeks to 1 year. Infertile women with both hypothyroidism and hyperprolactinemia also responded to treatment and their PRL levels returned to normal. Measurement of TSH and PRL should be done at early stage of infertility check up rather than straight away going for more costly tests or invasive procedures. Simple, oral hypothyroidism treatment for 3 months to 1 year can be of great benefit to conceive in otherwise asymptomatic infertile women.


Preparation and characterization of cyclodextrin nanosponges for organic toxic molecule removal.
Cyclodextrin-based nanosponges (CD-NS) are considered as safe and biocompatible systems for removing toxic molecules from the body. Rapid removal of toxic molecules that are formed in the body from certain food constituents, is relevant especially for patients affected by chronic kidney disease. Within the scope of this study, innovative cyclodextrin polymers were synthesized to form nanosponges able to remove indole, before it could form the toxic indoxyl sulfate in the body. Furthermore, in vivo studies were carried out using the two optimal CD-NS formulations by assessing physicochemical properties, stability, indole adsorption capacity and in vitro cytotoxicity. NS prepared from β-cyclodextrin cross-linked with toluene diisocyanate was found to be the most effective NS with an in vitro indole adsorption capacity of over 90%. In addition, this derivative was more stable in gastrointestinal media. Animal studies further revealed that oral CD-NSs did not tend to accumulate and damage gastrointestinal tissues and are excreted from the GI tract with minimal absorption. In conclusion, this study suggests that CD-NS formulations are effective and safe in removing toxic molecules from the body. Their potential use in veterinary or human medicine could reduce dialysis frequency and avoid hepatic and cardiac toxicity avoiding the indole formation.


Contraceptive counseling by general internal medicine faculty and residents.
Almost half of US pregnancies are unintended, resulting in many abortions and unwanted or mistimed births. Contraceptive counseling is an effective tool to increase patients' use of contraception. Using an online 20-item questionnaire, we evaluated the frequency of contraceptive counseling provided to reproductive-age women during a prevention-focused visit by University of Colorado internal medicine resident and faculty providers. We also evaluated factors hypothesized to affect contraceptive counseling frequency. Although more than 95% of the 146 medicine faculty and resident respondents agreed that contraceptive counseling is important, only one-quarter of providers reported providing contraceptive counseling "routinely" (defined as ≥80% of the time) to reproductive-age women during a prevention-focused visit. Providing contraceptive counseling routinely was strongly associated with taking an abbreviated sexual history "often"/"routinely" (odds ratio [OR]=11.6 [3.3 to 40.0]) and with high self-efficacy to provide contraceptive counseling (OR=6.5 [1.5 to 29.0]). However, fewer than two-thirds of providers reported taking an abbreviated sexual history "often"/"routinely." More than 70% of providers reported inadequate knowledge of contraceptive methods as a contraceptive counseling barrier. However, providers' perceived inadequate knowledge was not associated with traditional educational exposures, such as lectures and women's health electives. In prevention-focused visits with reproductive-age women, a minority of internal medicine faculty and residents reported routine contraceptive counseling. Future efforts to increase contraceptive counseling among internists should include interventions that increase provider contraceptive counseling self-efficacy and ensure that providers obtain an abbreviated sexual history.


A Novel Machine-Learning Framework Based on a Hierarchy of Dispute Models for the Identification of Fish Species Using Multi-Mode Spectroscopy.
Seafood mislabeling rates of approximately 20% have been reported globally. Traditional methods for fish species identification, such as DNA analysis and polymerase chain reaction (PCR), are expensive and time-consuming, and require skilled technicians and specialized equipment. The combination of spectroscopy and machine learning presents a promising approach to overcome these challenges. In our study, we took a comprehensive approach by considering a total of 43 different fish species and employing three modes of spectroscopy: fluorescence (Fluor), and reflectance in the visible near-infrared (VNIR) and short-wave near-infrared (SWIR). To achieve higher accuracies, we developed a novel machine-learning framework, where groups of similar fish types were identified and specialized classifiers were trained for each group. The incorporation of global (single artificial intelligence for all species) and dispute classification models created a hierarchical decision process, yielding higher performances. For Fluor, VNIR, and SWIR, accuracies increased from 80%, 75%, and 49% to 83%, 81%, and 58%, respectively. Furthermore, certain species witnessed remarkable performance enhancements of up to 40% in single-mode identification. The fusion of all three spectroscopic modes further boosted the performance of the best single mode, averaged over all species, by 9%. Fish species mislabeling not only poses health-related risks due to contaminants, toxins, and allergens that could be life-threatening, but also gives rise to economic and environmental hazards and loss of nutritional benefits. Our proposed method can detect fish fraud as a real-time alternative to DNA barcoding and other standard methods. The hierarchical system of dispute models proposed in this work is a novel machine-learning tool not limited to this application, and can improve accuracy in any classification problem which contains a large number of classes.


Understanding experiences of patients and family caregivers in the Mayo Clinic Care Transitions program: a qualitative study.
Care transitions programs are increasingly used to improve care and reduce re-admission of patients after hospitalization. To learn from the experience of patients who have participated in the Mayo Clinic Care Transitions (MCCT) program and to understand the patient experience, we sought perspectives of patients, caregivers, and providers who worked with participants of the MCCT program. Investigators interviewed 17 patients and nine of their caregivers about their experience with the MCCT program. Eight health care providers described provider experiences with the MCCT program. Data from semistructured interviews were audio recorded, transcribed, and evaluated through content analysis. Inductive coding methods were used to elicit themes about patient experience with the MCCT program. Patients, caregivers, and providers emphasized that the MCCT program prevented hospitalizations and contributed to the health and quality of life of participants. All three stakeholder groups emphasized the value of the home visit and provision of the visit on a patient's "home turf" as central to the program. Patients appreciated speaking to a provider without the stress and exertion of a trip to the clinic. Caregivers appreciated improved communication provided in the home visit and felt that home visits gave them peace of mind. Patients, caregivers, and providers also identified the need for improved phone triage and communication. Patients, caregivers, and providers acknowledged the care transitions problem and emphasized the benefits of seeing patients on their home turf rather than in an office visit. This qualitative study of patient, caregiver, and provider experiences further validates the importance of the MCCT program.


Association of diabetes insipidus, diabetes mellitus, optic atrophy, and deafness. The Wolfram or DIDMOAD syndrome.
Seven patients with a rare syndrome of diabetes insipidus (DI), diabetes mellitus (DM), optic atrophy (OA), neurosensory deafness (D), atony of the urinary tract, and other abnormalities (Wolfram or DIDMOAD syndrome) are reported. Of the seven patients, three siblings were followed up for 10-17 years. All seven patients had diabetes mellitus and optic atrophy; six had diabetes insipidus; and in the four patients investigated there was dilatation of the urinary tract. The severity of diabetes varied, and all required insulin for control of the hyperglycaemia. In one patient the course of the disease simulated maturity onset diabetes of the young; another presented with ketoacidosis; but none had haplotypes usually associated with insulin dependent diabetes mellitus. The diabetes insipidus responded to chlorpropamide, suggesting partial antidiuretic hormone deficiency. Onset of optic atrophy and loss of vision occurred relatively late and progressed slowly, although in one patient there was a rapid deterioration in visual acuity. Deafness was mild, of late onset, and of sensorineural origin. A degenerative process affecting the central and peripheral nervous system can explain all the manifestations of the syndrome except diabetes mellitus. The pathogenesis of the diabetes mellitus remains obscure.


Adhesion of composite to air-abraded enamel and dentin.
The purpose of this study was to compare the shear bond strength of a resin composite to air abraded and not air abraded enamel and dentin surfaces, and to evaluate the need for acid conditioning before the application of a single component adhesive. Enamel (E) and dentin (D) flat surfaces were obtained from a pool of recently extracted human teeth (30 incisors and 30 molars) and prepared with different treatments with and without the application of air abrasion (AA) and phosphoric acid (PA). Composite samples were bonded to the surfaces and stored in distilled water at 37 degrees C. After 24 hours, shear bond strength tests were carried out with an Instron machine and the results were subjected to statistical analysis. Maximum bond strength was obtained in the E + AA + PA group (34.74 MPa--2.70), followed by: E + PA (31.26 MPa--1.87); D + PA (24.09-1.72); D + AA + PA (20.61 MPa--1.79); D + AA (8.96 MPa--0.82) and E + AA (7.17 MPa--1.75). The results were significantly lower in those groups that were not acid conditioned (P < 0.01). The bond strength of composite to enamel was slightly increased as a result of the combination of AA with PA. However, the opposite effect was observed on dentin surfaces. Acid etching is a necessary clinical step even on air abraded surfaces.


Aggressive surgical resection after neoadjuvant chemoradiation therapy for locally advanced intrahepatic cholangiocarcinoma.
The poor prognosis of patients with intrahepatic cholangiocarcinoma (ICC) or hilar cholangiocarcinoma is well known. Herein, we described the first reported case of severe locally advanced ICC in which radical surgery was successfully achieved based on the marked effect of neoadjuvant chemoradiation therapy (NCRT) using gemcitabine. A 54-year-old man was admitted to our institution with obstructive jaundice. Abdominal computed tomography (CT) showed a large low-density mass in the caudate lobe, extensively involving the inferior vena cava and main portal vein. Moreover, nodal involvements of the hepatoduodenal ligament were detected concurrently. We therefore regarded this tumor as a severe locally advanced ICC and attempted to initiate combined treatment with gemcitabine (800 mg/m(2) biweekly) and three-dimensional conformation radiation (45 Gy/25 days). After completion of NCRT, this patient underwent a left trisegmentectomy with combined resection of the portal vein and inferior vena cava. Postoperative microscopic findings surprisingly revealed that more than 90% of tumor cells had disappeared with extensive fibrosis, achieving tumor downstaging and tumor volume reduction which were related to the radical resection. In conclusion, ICC showed a favorable histological response to chemoradiation therapy using gemcitabine. Further studies are needed to conclusively assess the effect of NCRT on locally advanced ICC.


Genetic control of the antibody response. I. Demonstration of determinant-specific differences in response to synthetic polypeptide antigens in two strains of inbred mice.
Immunization of CBA and C57 mice with a branched, multichain synthetic polypeptide, poly (tyr,glu)-poly DL-ala--poly lys, ((T,G)-A--L), in Freund's complete adjuvant results in a tenfold or more difference in the antigen-binding capacity of sera from the two strains, although they respond equally to bovine serum albumin. Immunization of CBA x C57 F(1), F(1) x CBA, and F(1) x C57 mice reveals definite genetic control of the response to (T,G)-A--L, which appears to be due to a single major genetic factor, with perhaps one or more modifying factors. Immunization of CBA and C57 mice with (H,G)-A--L, a synthetic polypeptide in which histidine replaces tyrosine, gives the opposite result, CBA's respond and C57's do not. From this, it appears that the genetic control of the response to (T,G)-A--L is specific for the antigenic determinant. The implications of these results are discussed.


LC/MS analysis of tetrodotoxin and its deoxy analogs in the marine puffer fish Fugu niphobles from the southern coast of Korea, and in the brackishwater puffer fishes Tetraodon nigroviridis and Tetraodon biocellatus from Southeast Asia.
Tetrodotoxin (TTX) and its deoxy analogs, 5-deoxyTTX, 11-deoxyTTX, 6,11-dideoxyTTX, and 5,6,11-trideoxyTTX, were quantified in the tissues of three female and three male specimens of the marine puffer fish, Fugu niphobles, from the southern coast of Korea, and in the whole body of the brackishwater puffer fishes, Tetraodon nigroviridis (12 specimens) and Tetrodon biocellatus (three specimens) from Southeast Asia using LC/MS in single ion mode (SIM). Identification of these four deoxy analogs in the ovarian tissue of F. niphobles were further confirmed by LC/MS/MS. TTX and 5,6,11-trideoxyTTX were detected in all three puffer fish species as the major TTX analogs, similar to Japanese Fugu pardalis. While 6,11-dideoxyTTX was also found to be a major analog in almost all tissues of Korean F. niphobles, this analog was minor in the two Tetraodon species and Japanese F. pardalis. Among the tissues of F. niphobles, the concentrations of TTXs were highest in the ovaries (female) and skin (female and male).


Effects of varenicline on lung tissue in the animal model.
This study aimed to investigate acute and chronic effects of varenicline on lung tissue in an experimental study. A total of 34 rats were randomly allocated into study (varenicline) and control groups. The rats were divided into two groups (i) control group, (ii) varenicline group. Then, the rats in the each group were sub-divided equally in turn as acute (C1; V1) and chronic (C2; V2) ; all rats of acute and chronic groups were sacrificed under the anesthesia on the 45th day for acute group [C1 (n=5) and V1 (n=12)] and the 90th day for chronic group [C2 (n=5) and V2 (n=12)], respectively. Thus, biochemical and histopathological analysis were carried out. Thirty four rats completed the study, 24 were in varenicline group and 10 were in control group. In chronic exposure to varenicline, oxidant levels comprising of malondialdehyde (MDA), and myeloperoxidase (MPO) increased and superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and glutathione peroxidase (GPx) levels, named as antioxidants, decreased significantly when compared to the control group. MDA and MPO levels were also significantly higher and SOD, CAT, GPx, GSH levels were also significantly lower in chronic varenicline group when compared to acute varenicline group. These findings were also supported by histopathological observations. This is the first study, which evaluated pulmonary effects of varenicline experimentally on an animal model. It was observed that chronic varenicline treatments cause inflammation and lung cell injury.