Variations in maternal care alter corticosterone and 17beta-estradiol levels, estrous cycle and folliculogenesis and stimulate the expression of estrogen receptors alpha and beta in the ovaries of UCh rats.
Variations in maternal care are associated with neonatal stress, hormonal disturbances and reproductive injuries during adulthood. However, the effects of these variations on sex hormones and steroid receptors during ovary development remain undetermined. This study aimed to investigate whether variations in maternal care are able to influence the hormonal profile, follicular dynamics and expression of AR, ER-alpha and ER-beta in the ovaries of UCh rat offspring. Twenty-four adult UCh rats, aged 120 days, were randomly divided into two groups (UChA and UChB) and mated. Maternal care was assessed from birth (day 0) to the 10th postnatal day (PND). In adulthood, twenty adult female rats (UChA and UChB offspring; n = 10/group), aged 120 days, were euthanized by decapitation during the morning estrus. UChA females (providing high maternal care) more frequently displayed the behaviors of carrying pups, as well as licking/grooming and arched back nursing cares. Also, mothers providing high care had elevated corticosterone levels. Additionally, offspring receiving low maternal care showed the highest estrous cycle duration, increased corticosterone and 17beta-estradiol levels, overexpression of receptors ER-alpha and ER-beta, increased numbers of primordial, antral and mature follicles and accentuated granulosa cell proliferation. Our study suggests that low maternal care alters corticosterone and 17beta-estradiol levels, disrupting the estrous cycle and folliculogenesis and differentially regulating the expression of ER-alpha and ER-beta in the ovaries of adult rats.


Using electronic health records to conduct children's health insurance surveillance.
Health insurance options are changing. Electronic health record (EHR) databases present new opportunities for providers to track the insurance coverage status of their patients. This study demonstrates the use of EHR data for this purpose. Using EHR data from the OCHIN Network of community health centers, we conducted a retrospective cohort study of data from children presenting to a community health center in 2010-2011 (N = 185,959). We described coverage patterns for children, used generalized estimating equation logistic regression to compare uninsured children with those with insurance, and assessed insurance status at subsequent visits. At their first visit during the study period, 21% of children had no insurance. Among children uninsured at a first visit, 30% were uninsured at all subsequent visits. In multivariable analyses (including gender, age, race, ethnicity, language, income, location, and type of clinic), we observed significant differences in the characteristics of children who were uninsured as compared with those with insurance coverage. For example, compared with white, non-Hispanic children, nonwhite and/or Hispanic children had lower odds of being uninsured than having Medicaid/Medicare (adjusted odds ratio, 0.73; 95% confidence interval: 0.71-0.75) but had higher odds of being uninsured than having commercial insurance (adjusted odds ratio, 1.50; 95% confidence interval: 1.44-1.56). Nearly one-third of children uninsured at their first visit remained uninsured at all subsequent visits, which suggests a need for clinics to conduct insurance surveillance and develop mechanisms to assist patients with obtaining coverage. EHRs can facilitate insurance surveillance and inform interventions aimed at helping patients obtain and retain coverage.


Meta-Analyses of Genome-Wide Association Studies for Postpartum Depression.
Postpartum depression (PPD) is a common subtype of major depressive disorder (MDD) that is more heritable, yet is understudied in psychiatric genetics. The authors conducted meta-analyses of genome-wide association studies (GWASs) to investigate the genetic architecture of PPD. Meta-analyses were conducted on 18 cohorts of European ancestry (17,339 PPD cases and 53,426 controls), one cohort of East Asian ancestry (975 cases and 3,780 controls), and one cohort of African ancestry (456 cases and 1,255 controls), totaling 18,770 PPD cases and 58,461 controls. Post-GWAS analyses included 1) single-nucleotide polymorphism (SNP)-based heritability ([Formula: see text]), 2) genetic correlations between PPD and other phenotypes, and 3) enrichment of the PPD GWAS findings in 27 human tissues and 265 cell types from the mouse central and peripheral nervous system. No SNP achieved genome-wide significance in the European or the trans-ancestry meta-analyses. The [Formula: see text] of PPD was 0.14 (SE=0.02). Significant genetic correlations were estimated for PPD with MDD, bipolar disorder, anxiety disorders, posttraumatic stress disorder, insomnia, age at menarche, and polycystic ovary syndrome. Cell-type enrichment analyses implicate inhibitory neurons in the thalamus and cholinergic neurons within septal nuclei of the hypothalamus, a pattern that differs from MDD. While more samples are needed to reach genome-wide levels of significance, the results presented confirm PPD as a polygenic and heritable phenotype. There is also evidence that despite a high correlation with MDD, PPD may have unique genetic components. Cell enrichment results suggest GABAergic neurons, which converge on a common mechanism with the only medication approved by the U.S. Food and Drug Administration for PPD (brexanolone).


Recognition of Pneumocystis carinii antigen on its surface by immunohistochemistry and immunoelectron microscopy.
The aim of this study was to detect the surface antigens in different stages of experimental induced Pneumocystis carinii in Sprague-Dawley rats. Immunohistochemical staining with monoclonal (900, 902 and 904) and polyclonal (SP-D) antibodies demonstrated that the P. carinii organisms were mostly in the alveolar lumina. The binding sites of the monoclonal (900, 902 and 904) and polyclonal (SP-D) antibodies developed against P. carinii were examined at the ultrastructural level by using a post-embedding immunogold labeling. The gold particles were observed evenly on the surface of precyst and cyst stages of the P. carinii. In the trophozoite stage, scattered gold particles were seen on the pellicles and tubular expansions. The monoclonal antibodies reacted mainly with pellicles of P. carinii, whereas SP-D labeled pellicles, intracystic bodies, cytoplasms of alveolar macrophages, free floating surfactant material in the alveolar spaces, and adjacent type II epithelial cells. In the immunogold labeling, basically no significant differences were found in the precyst, cyst, and ruptured cyst stages. These results indicate that the gold particles were observed adhering to every stage of P. carinii, mostly concentrated on the pellicles, and more concentrated in the precyst or cyst stage than trophozoite stage which may be due to an increase in antigen accumulation during development from the trophozoite to the cyst.


Neurological adverse effects of methylphenidate may be misdiagnosed as meningoencephalitis.
We present a case of adverse neurological effects of methylphenidate therapy for attention deficit and hyperactivity disorder (ADHD). A 7-year-old boy presented to the emergency department (ED) having developed ataxic gait, orofacial dyskinesias and choreoathetosis of the limbs. The results of all blood investigations, EEG and CT scan of the head were unremarkable. Subsequently, a detailed history revealed he was being treated for ADHD, being started on methylphenidate in the past 3 months. Discontinuation of methylphenidate led to significant and rapid amelioration of neurological adverse effects.


Oritavancin: a new opportunity for outpatient therapy of serious infections.
Oritavancin is a new antibiotic for the treatment of serious infections with Gram-positive bacteria. It has been shown to be effective against methicillin-susceptible and -resistant Staphylococcus aureus as well as enterococci. With a terminal half-life of 393 hours, oritavancin lends itself to a convenient and potentially cost-effective single-dose regimen. The single-dose regimen is currently being evaluated in pivotal phase 3 studies. This unique property provides an opportunity to assure consistent, effective, and safe treatment for serious infections while reducing the costs of care through the elimination of multiple infusions, reduced medical care staff, shorter hospital stays, and avoidance of hospital-acquired infections. These features seem ideal for the use of oritavancin in the outpatient management of serious infections. The impact that oritavancin will have on outpatient therapy is unclear. Current models will need to change with only a single infusion. Physician monitoring of the infection and underlying diseases may not be as frequent despite the need for close follow-up and frequent evaluations. There will be less need for a team of outpatient infusion specialists. Outpatient therapy will be compensated less without multiple infusions. With the possibility of fewer physician and other medical visits, there will be more responsibility for the patient and family and a reliance on patients to care for themselves. Although oritavancin offers tremendous theoretical advantages in the outpatient treatment of serious infections, care should be taken to assure the quality of care through changes in reimbursement, patient education, and development of systems to monitor care and outcomes.


N2-(1-carboxyethyl)methionine. A 'pseudo-opine' in octopine-type crown-gall tumours.
A novel methionine-containing plasmid-determined compound, N2-(1-carboxyethyl)methionine (NCEM) has been identified in crown-gall tumours induced by octopine-type strains of Agrobacterium tumefaciens. NCEM is probably synthesized by octopine synthase. Cell-free preparations from octopine-type strains of A. tumefaciens can degrade NCEM; however, the bacterium cannot transport the compound into the cell, although these strains can take up and degrade the octopine family of opines.


Life cycle of Leishmania major (Kinetoplastida: Trypanosomatidae) in the neotropical sand fly Lutzomyia longipalpis (Diptera: Psychodidae).
The development of Leishmania major Yakimoff & Schokhor in the New World sand fly Lutzomyia longipalpis (Lutz & Neiva) was examined by light and electron microscopy. In this unnatural host, parasites differentiated into 10 typical morphological forms, multiplied at three sites, migrated anteriorly and established in the foregut, and attached to gut surfaces. In the blood meal, amastigotes divided and transformed into two successive dividing, stumpy promastigote stages. Elongate nectomonad promastigotes developed from stumpy forms and subsequently rounded up in some flies into paramastigotes and opisthomastigotes. Differentiation into round opisthomastigotes and the apparent fusion of paramastigotes in the blood meal were novel observations in this study. Three nectomonad promastigotes--elongate, short, and metacyclic--were free-swimming in the midgut lumen. Elongate nectomonad promastigotes were highly oriented in the midgut, with their flagella embedded between the epithelial microvilli. Short haptomonad promastigotes were the predominant form attached to the intima of the stomodeal valve, whereas pear-shaped haptomonad promastigotes and paramastigotes colonized surfaces of the esophagus and pharynx. Peripylarian attachment of promastigotes and paramastigotes in the pylorus, ileum, and colon was noted in 21% of flies, suggesting that suprapylarian leishmanias have not lost the ability to colonize the hindgut. L. longipalpis was a successful biological host for L. major, allowing complete development of the parasite.


Increased concentration of glucocorticoid receptors in rabbit iris--ciliary body compared to rabbit liver.
Dexamethasone binding to both rabbit iris-ciliary body and rabbit liver glucocorticoid receptors was compared by means of a batch assay in which the binding components were adsorbed to hydroxylapatite. The results showed that homogenates of both the rabbit iris-ciliary body and the rabbit liver contained a single class of high-affinity receptors. Binding affinity was virtually identical for the two receptors. A comparison of total binding sites in homogenates of the iris-ciliary body and the liver revealed that the iris-ciliary body glucocorticoid receptor was present in nearly twice the concentration of the hepatic receptor.


Relationship between levels of very virulent MDV in poultry dust and in feather tips from vaccinated chickens.
To assess the effect of various vaccine strains on replication and shedding of virulent Marek's disease virus from experimentally infected chickens, quantitative PCR (q-PCR) methods were developed to accurately quantify viral DNA in infected chickens and in the environment in which they were housed. Four groups of 10 chickens, kept in poultry isolators, were vaccinated at 1 day old with one of four vaccines covering each of the three vaccine serotypes, then challenged with very virulent MDV strain Md5 at 8 days of age. At regular time-points, feather tips were collected from each chicken and poultry dust was collected from the air-extract prefilter of each isolator. DNA was extracted from feather and dust samples and subjected to real-time q-PCR, targeting the U(S)2 gene of MDV-1, in order to measure Md5 level per 10(4) feather tip cells or per microgram of dust. Accuracy of DNA extraction from dust and real-time q-PCR were validated by comparing either q-PCR cycle threshold values or the calculated MDV genome level; for use in q-PCR, DNA was extracted from serial dilutions of MDV-infected dust diluted with noninfected dust, or DNA from MDV-infected dust was diluted with DNA from noninfected dust. The results confirmed the accuracy and sensitivity of dust DNA extraction and subsequent q-PCR and showed that differences in virus levels between dust samples truly reflect differences in shedding. Vaccination delayed both replication of Md5 in feather tips and shedding of Md5. First detection of Md5 in feather tips always preceded or coincided with first detection in dust in each group. pCVI988 and HVT+SB-1 were the most efficient vaccines in reducing both replication and shedding of Md5. There was close correlation between mean virus level in feathers of each group and mean virus level in the dust shed by that group. This relationship was similar in each of the vaccinated groups, demonstrating that measurement of the virus in dust can be used to monitor accurately both the infection status of the chickens and environmental contamination by MDV.


Arrhythmia-induced cardiomyopathies: the riddle of the chicken and the egg still unanswered?
The hypothesis testing of inappropriate fast, irregular, or asynchronous myocardial contraction provoking cardiomyopathy has been the primary focus of numerous research efforts, especially during the last few decades. Rapid ventricular rates resulting from supraventricular arrhythmias and atrial fibrillation (AF), irregularity of heart rhythm-basic element of AF-and asynchrony, as a consequence of right ventricular pacing, bundle branch block, or frequent premature ventricular complexes, have been established as primary causes of arrhythmia-induced cardiomyopathy. The main pathophysiological pathways involved have been clarified, including neurohumoral activation, energy stores depletion, and abnormalities in stress and strain. Unfortunately, from a clinical point of view, patients usually seek medical advice only when symptoms develop, while the causative arrhythmia may be present for months or years, resulting in myocardial remodelling, diastolic, and systolic dysfunction. In some cases, making a definite diagnosis may become a strenuous exercise for the treating physician, as the arrhythmia may not be present and, additionally, therapy must be applied for the diagnosis to be confirmed retrospectively. The diagnostic process is also hardened due to the fact that strict diagnosing criteria are still a matter of discrepancy. Therapy options include pharmaceutical agents trials, catheter-based therapies and, in the context of chronic ventricular pacing, resynchronization. For the majority of patients, partial or complete recovery is expected, although they have to be followed up thoroughly due to the risk of recurrence. Large, randomized controlled trials are more than necessary to optimize patients' stratification and therapeutic strategy choices.


Topologically associating domains and their role in the evolution of genome structure and function in Drosophila.
Topologically associating domains (TADs) were recently identified as fundamental units of three-dimensional eukaryotic genomic organization, although our knowledge of the influence of TADs on genome evolution remains preliminary. To study the molecular evolution of TADs in Drosophila species, we constructed a new reference-grade genome assembly and accompanying high-resolution TAD map for D. pseudoobscura Comparison of D. pseudoobscura and D. melanogaster, which are separated by ∼49 million years of divergence, showed that ∼30%-40% of their genomes retain conserved TADs. Comparative genomic analysis of 17 Drosophila species revealed that chromosomal rearrangement breakpoints are enriched at TAD boundaries but depleted within TADs. Additionally, genes within conserved TADs show lower expression divergence than those located in nonconserved TADs. Furthermore, we found that a substantial proportion of long genes (>50 kbp) in D. melanogaster (42%) and D. pseudoobscura (26%) constitute their own TADs, implying transcript structure may be one of the deterministic factors for TAD formation. By using structural variants (SVs) identified from 14 D. melanogaster strains, its three closest sibling species from the D. simulans species complex, and two obscura clade species, we uncovered evidence of selection acting on SVs at TAD boundaries, but with the nature of selection differing between SV types. Deletions are depleted at TAD boundaries in both divergent and polymorphic SVs, suggesting purifying selection, whereas divergent tandem duplications are enriched at TAD boundaries relative to polymorphism, suggesting they are adaptive. Our findings highlight how important TADs are in shaping the acquisition and retention of structural mutations that fundamentally alter genome organization.


Pricklier with the proper predator? Predator-induced small-scale changes of spinescence in Daphnia.
Phenotypic plasticity in defensive traits is a common response of prey organisms to variable and unpredictable predation regimes and risks. Cladocerans of the genus Daphnia are keystone species in the food web of lentic freshwater bodies and are well known for their ability to express a large variety of inducible morphological defenses in response to invertebrate and vertebrate predator kairomones. The developed defenses render the daphnids less susceptible to predation. So far, primarily large-scale morphological defenses, like helmets, crests, and tail-spines, have been documented. However, less is known on whether the tiny spinules, rather inconspicuous traits which cover many Daphnia's dorsal and ventral carapace margins, respond to predator kairomones, as well. For this reason, we investigated two Daphnia species (D. magna and D. longicephala) concerning their predator kairomone-induced changes in dorsal and ventral spinules. Since these small, inconspicuous traits may only act as a defense against predatory invertebrates, with fine-structured catching apparatuses, and not against vertebrate predators, we exposed them to both, an invertebrate (Triops cancriformis or Notontecta maculata) and a vertebrate predator (Leucaspius delineatus). Our results show that the length of these spinules as well as spinules-covered areas vary, likely depending on the predator the prey is exposed to. We further present first indications of a Daphnia species-specific elongation of the spinules and an increase of the spinules-bearing areas. Although we cannot exclude that spinescence is altered because it is developmentally connected to changes in body shape in general, our results suggest that the inducible alterations to the spinule length and spinules-covered areas disclose another level of predator-induced changes in two common Daphnia species. The predator-induced changes on this level together with the large-scale and ultrastructural defensive traits may act as the overall morphological defense, adjusted to specific predator regimes in nature.


Joint Sparse Representation and Embedding Propagation Learning: A Framework for Graph-Based Semisupervised Learning.
In this paper, we propose a novel graph-based semisupervised learning framework, called joint sparse representation and embedding propagation learning (JSREPL). The idea of JSREPL is to join EPL with sparse representation to perform label propagation. Like most of graph-based semisupervised propagation learning algorithms, JSREPL also constructs weights graph matrix from given data. Different from classical approaches which build weights graph matrix and estimate the labels of unlabeled data in sequence, JSREPL simultaneously builds weights graph matrix and estimates the labels of unlabeled data. We also propose an efficient algorithm to solve the proposed problem. The proposed method is applied to the problem of semisupervised image clustering using the ORL, Yale, PIE, and YaleB data sets. Our experiments demonstrate the effectiveness of our proposed algorithm.


The clinical significance of perivalvular pannus in prosthetic mitral valves: Can cardiac CT be helpful?
The clinical significance of pannus in the prosthetic mitral valve (MV) is not well documented. To investigate the clinical significance of pannus on cardiac computed tomography (CT) in patients with a prosthetic MV. A total of 130 patients with previous MV replacement who underwent cardiac CT were retrospectively included in this study. The presence of pannus, paravalvular leak (PVL) around the prosthetic MV and limitation of motion (LOM) of the MV were analyzed using CT. Between patients with MV pannus and those without pannus, CT, echocardiographic, and redo-surgery findings were compared. The diagnostic performance of CT and transesophageal echocardiography (TEE) for the detection of MV pannus was also compared, using surgical findings as a standard reference. MV pannus was observed on cardiac CT in 32.3% of the study population. Patients with MV pannus detected on CT more commonly had LOM (28.2% vs. 15.2%) and less frequently had PVL of the prosthetic MV (16.7% vs. 25%) than patients without MV pannus (P>0.05). Prosthetic valve obstruction (PVO) due prosthetic MV pannus requiring redo-surgery was present in only five patients (11.9%). Cardiac CT detected MV pannus with sensitivity of 65.2% and specificity of 80.9% and showed better diagnostic performance than TEE (P<0.05). Prosthetic MV pannus can frequently be seen on cardiac CT. However, its clinical significance should be assessed with careful consideration, because PVO due to MV pannus is relatively uncommon, and pannus can be seen in patients without any clinical problems.


An antibody to VacA of Helicobacter pylori in cerebrospinal fluid from patients with Guillain-Barre syndrome.
To detect antibodies to recombinant vacuolating cytotoxin (r-VacA) of Helicobacter pylori in cerebrospinal fluid (CSF) from patients with Guillain-Barre syndrome (GBS). CSF samples from 13 patients with GBS (electrophysiologically classified as eight acute inflammatory demyelinating polyradiculoneuropathy (AIDP), four acute motor axonal neuropathy (AMAN), and one unexcitable nerve conduction) and eight disease control patients were studied. The r-VacA protein was separated by SDS/PAGE, and Western blot analysis was carried out. Six of the 13 patients with GBS had a specific IgG antibody to VacA of H pylori, which was confirmed by absorption experiments using r-VacA. Every patient with positive CSF anti-r-VacA IgG had AIDP. The sequence homology previously found between VacA and human (Na(+)+K(+))-ATPase A subunit suggests that antibodies to VacA involve ion channels in abaxonal Schwann cell plasmalemma resulting in demyelination in some patients with GBS.


A study on using texture analysis methods for identifying lobar fissure regions in isotropic CT images.
The major hurdle for segmenting lung lobes in computed tomographic (CT) images is to identify fissure regions, which encase lobar fissures. Accurate identification of these regions is difficult due to the variable shape and appearance of the fissures, along with the low contrast and high noise associated with CT images. This paper studies the effectiveness of two texture analysis methods - the gray level co-occurrence matrix (GLCM) and the gray level run length matrix (GLRLM) - in identifying fissure regions from isotropic CT image stacks. To classify GLCM and GLRLM texture features, we applied a feed-forward back-propagation neural network and achieved the best classification accuracy utilizing 16 quantized levels for computing the GLCM and GLRLM texture features and 64 neurons in the input/hidden layers of the neural network. Tested on isotropic CT image stacks of 24 patients with the pathologic lungs, we obtained accuracies of 86% and 87% for identifying fissure regions using the GLCM and GLRLM methods, respectively. These accuracies compare favorably with surgeons/radiologists' accuracy of 80% for identifying fissure regions in clinical settings. This shows promising potential for segmenting lung lobes using the GLCM and GLRLM methods.


Effects of nociceptin/orphanin FQ in the acquisition of contextual and tone fear conditioning in rats.
Nociceptin, or orphanin FQ (N/OFQ), the endogenous ligand of NOP receptors, is known to regulate learning and memory processes. To verify the role of N/OFQ in the acquisition of contextual (CFC) and tone fear conditioning (TFC), Wistar male rats received intracerebroventricular injections of N/OFQ (0.1-5.0 nmol) before training, and were tested 24 and 48 hr later to access the freezing response to context and tone, respectively. The intermediate doses (1.0 and 2.5 nmol) impaired the CFC test, sparing TFC. The highest dose (5.0 nmol) reduced freezing during both tests, a result that may be due to nonspecific effects. The posttraining injection of N/OFQ (1 or 5 nmol) did not interfere with CFC and TFC, suggesting a specific effect of the peptide in acquisition processes. Moreover, the impairment observed with N/OFQ (1 nmol) in CFC cannot be attributed to a state-dependent learning because it was not reversed by its pretest administration. The data support the negative role of N/OFQ in the acquisition of aversively motivated tasks, which encompass a spatial component and depend on the hippocampus.


Genomic analysis of cardiac surgery-associated Mycobacterium chimaera infections in Italy.
One hundred and twenty-two Mycobacterium chimaera strains isolated in Italy from cardiac surgery-related patients, cardiac surgery-unrelated patients and from heater-cooler units, were submitted to whole-genome sequencing and to subsequent SNP analysis. All but one strains isolated from cardiac surgery-related patients belonged to Subgroup 1.1 (19/23) or Subgroup 1.8 (3/23). Only 28 out of 79 strains isolated from heater-cooler units belonged to groupings other than 1.1 and 1.8. The strains isolated from cardiac surgery-unrelated patients were instead distributed across the phylogenetic tree. Our data, the first on isolates from Italy, are in agreement with a recent large genomic study suggesting a common source, represented by strains belonging to Subgroups 1.1 and 1.8, of cardiac surgery-related Mycobacterium chimaera infections. The strains belonging to groupings other than 1.1 and 1.8 isolated from heather-cooler units evidently resulted from contaminations at hospital level and had no share in the Mycobacterium chimaera outbreak. One Mycobacterium chimaera strain investigated in this study proved distant from every previously known Mycobacterium chimaera Groups (1, 2, 3 and 4) and we propose to assign to a novel group, named "Group 5".


A methodological procedure for evaluating the impact of hemolysis on circulating microRNAs.
Circulating microRNAs (miRNAs) are promising non-invasive biomarkers whose expression may be affected by confounding factors, including hemolysis, that should be considered in studies of miRNA discovery. The present study proposes a methodology for evaluating the impact of hemolysis on the expression of miRNAs. An experiment of in vitro controlled hemolysis was designed for assessing if changes in the expression of eight miRNAs observed to be circulating in plasma may be associated with hemolysis, and also to estimate the level of red blood cell (RBC) contamination in plasma samples where the expression of these miRNAs will be measured. It was confirmed that four miRNAs, miR-16, miR-92a, miR-451 and miR-486, known to be present in blood cells, were influenced by contamination of RBCs. Furthermore, it was demonstrated that miR-378 and miR-30c are hemolysis-independent and that the expression of miR-320 and miR-324-3p was associated with the level of RBC contamination. This procedure is proposed as a tool for the evaluation of the influence of hemolysis on candidate circulating miRNA biomarkers prior to their analysis in plasma samples.


Gangliosides expressed by the renal cell carcinoma cell line SK-RC-45 are involved in tumor-induced apoptosis of T cells.
It is now understood that the genetic plasticity of cancer cells can lead to alterations that confer selective growth advantages to the tumor, some of which play a role in immune escape. A number of mutations veiling tumor cells from host immune defenses have been well characterized but more recent studies suggest that a variety of tumors can also express products that are actually toxic for the immune effectors. A component of this tumor-induced T-cell death has been attributed to receptor-mediated apoptosis. Some tumors, however, synthesize soluble factors that mediate similar effects. In this regard, we previously showed that supernatants from explanted renal cell carcinoma (RCC) tumors sensitized normal T cells to activation induced cell death, and the responsible products had the features of gangliosides. We have also shown that renal tumor lines, including SK-RC-45, induce apoptosis of both Jurkat cells and normal T lymphocytes. Here, we used the ganglioside synthesis inhibitor PPPP to define the role of gangliosides in RCC cell line (SK-RC-45)-mediated T cell and Jurkat cell apoptosis and to elucidate the proapoptotic molecular events by which the glycosphingolipids produce their effects. The ganglioside-synthesizing SK-RC-45 line stimulated the TUNEL (terminal deoxynucleotidyl transferase-mediated nick end labeling) positivity of cocultured T cells by a mechanism that involved decreasing lymphocyte expression levels of Bcl-2 and Bcl-(XL), inducing cytochrome c release from their mitochondria and activating caspases 9 and 3. These proapoptotic events were partially or completely abrogated when tumor cells were pretreated with PPPP for 5 days before the SK-RC-45/T lymphocyte coincubation, a regimen that reduced tumor-associated ganglioside levels by 70-80%. Our results suggest that gangliosides may be key mediators of RCC-induced T-cell apoptosis and imply that they contribute to the T-cell dysfunction in the tumor microenvironment.


Osteoblasts produce monocyte chemoattractant protein-1 in a murine model of Staphylococcus aureus osteomyelitis and infected human bone tissue.
Incidences of osteomyelitis caused by Staphylococcus aureus have increased dramatically in recent years, in part, due to the appearance of community-acquired antibiotic-resistant strains. Therefore, understanding the pathogenesis of this organism has become imperative. Recently, we have described the surprising ability of bone-forming osteoblasts to secrete a number of important immune mediators when exposed to S. aureus in vitro. In the present study, we provide the first evidence for the in vivo production of the pivotal inflammatory chemokine, monocyte chemoattractant protein-1 (MCP-1), by osteoblasts during S. aureus-associated bone infection. Quantitative real-time PCR was employed to determine levels of mRNA encoding MCP-1 in vivo using a mouse model that closely resembles the pathology of trauma-induced staphylococcal osteomyelitis. Expression of this inflammatory chemokine and osteoblast-specific markers was investigated by confocal laser scanning microscopy in bone tissue from organ cultures of neonatal mouse calvaria and from the in vivo mouse model. Furthermore, the clinical relevancy of these findings was investigated by performing similar studies on infected human bone tissue from patients with S. aureus-associated osteomyelitis. Here, we confirm that expression of mRNA encoding MCP-1 is elevated in bacterially infected murine bone tissue. Importantly, we show that these increases translate into marked elevations in the expression of MCP-1 protein that co-localizes with osteoblast markers in infected bone tissue. Such increases could not be attributed solely to mechanical damage as a similar response was observed in infected but otherwise undamaged organ cultures. Finally, we have demonstrated the in vivo production of MCP-1 by osteoblasts in bone specimens from patients with S. aureus-associated osteomyelitis. As such, these studies demonstrate that bacterial challenge of osteoblasts during bone diseases such as staphylococcal osteomyelitis induces cells to produce a key inflammatory chemokine that can direct appropriate host responses or may contribute to progressive inflammatory damage.


Reactivity assessment of chalcones by a kinetic thiol assay.
The electrophilic nature of chalcones (1,3-diphenylprop-2-en-1-ones) and many other α,β-unsaturated carbonyl compounds is crucial for their biological activity, which is often based on thiol-mediated regulation processes. To better predict their biological activity a simple screening assay for the assessment of the second-order rate constants (k(2)) in thia-Michael additions was developed. Hence, a clear structure-activity relationship of 16 differentially decorated hydroxy-alkoxychalcones upon addition of cysteamine could be established. Moreover, amongst other naturally occurring α,β-unsaturated carbonyl compounds k(2) values for curcumin and cinnamaldehyde were gained while cinnamic acids or esters gave no or very slow reactions.


Genetic profile of an Oka varicella vaccine virus variant isolated from an infant with zoster.
Varicella virus vaccine strain Oka (V-Oka) has in rare cases caused zoster in vaccinated people. Despite broad usage of V-Oka, little is known about varicella-zoster virus genomic sequence variation of strains in vaccine and isolates from patients with vaccine adverse events. Direct sequencing of 20 regions of V-Oka-GSK was compared to the sequences of the original V-Oka-Biken, GlaxoSmithKline Oka vaccine (V-Oka-GSK), and Oka-parental (P-Oka) strains. We analyzed single nucleotide polymorphisms (SNP) differentiating the Oka parental and Oka vaccine strains identified in open reading frames (ORFs) 6, 9A, 10, 21, 31, 39, 50, 51, 52, 54, 55, and 59 and eight base substitutions within ORF 62. Sixteen of these SNP impose an amino acid change in the corresponding gene product. The genotypic analysis revealed that (i) both V-Oka-GSK and V-Oka-Biken comprise mixtures of strains represented in variable proportion from lot to lot; (ii) V-Oka-GSK/zoster isolated from the zoster patient had six wild-type SNP in ORF 9A, 10, 21, 52, 55, and 62 (mutation 108838); (iii) none of the six revertant SNP would reliably discriminate Oka vaccine from the wild type; and (iv) the genomic variation found in V-Oka/zoster might be associated with changes in the biological behavior of the virus. Further studies will be needed to identify potential virulence factors in variant vaccine strains.


Impact of FDG-PET on radiation therapy volume delineation in non-small-cell lung cancer.
Locoregional failure remains a significant problem for patients receiving definitive radiation therapy alone or combined with chemotherapy for non-small-cell lung cancer (NSCLC). Positron emission tomography (PET) with [(18)F]fluoro-2-deoxy-d-glucose (FDG) has proven to be a valuable diagnostic and staging tool for NSCLC. This prospective study was performed to determine the impact of treatment simulation with FDG-PET and CT on radiation therapy target volume definition and toxicity profiles by comparison to simulation with computed tomography (CT) scanning alone. Twenty-six patients with Stages I-III NSCLC were studied. Each patient underwent sequential CT and FDG-PET simulation on the same day. Immobilization devices used for both simulations included an alpha cradle, a flat tabletop, 6 external fiducial markers, and a laser positioning system. A radiation therapist participated in both simulations to reproduce the treatment setup. Both the CT and fused PET/CT image data sets were transferred to the radiation treatment planning workstation for contouring. Each FDG-PET study was reviewed with the interpreting nuclear radiologist before tumor volumes were contoured. The fused PET/CT images were used to develop the three-dimensional conformal radiation therapy (3DCRT) plan. A second physician, blinded to the results of PET, contoured the gross tumor volumes (GTV) and planning target volumes (PTV) from the CT data sets, and these volumes were used to generate mock 3DCRT plans. The PTV was defined by a 10-mm margin around the GTV. The two 3DCRT plans for each patient were compared with respect to the GTV, PTV, mean lung dose, volume of normal lung receiving > or =20 Gy (V20), and mean esophageal dose. The FDG-PET findings altered the AJCC TNM stage in 8 of 26 (31%) patients; 2 patients were diagnosed with metastatic disease based on FDG-PET and received palliative radiation therapy. Of the 24 patients who were planned with 3DCRT, PET clearly altered the radiation therapy volume in 14 (58%), as follows. PET helped to distinguish tumor from atelectasis in all 3 patients with atelectasis. Unsuspected nodal disease was detected by PET in 10 patients, and 1 patient had a separate tumor focus detected within the same lobe of the lung. Increases in the target volumes led to increases in the mean lung dose, V20, and mean esophageal dose. Decreases in the target volumes in the patients with atelectasis led to decreases in these normal-tissue toxicity parameters. Radiation targeting with fused FDG-PET and CT images resulted in alterations in radiation therapy planning in over 50% of patients by comparison with CT targeting. The increasing availability of integrated PET/CT units will facilitate the use of this technology for radiation treatment planning. A confirmatory multicenter, cooperative group trial is planned within the Radiation Therapy Oncology Group.


Protein sectors: statistical coupling analysis versus conservation.
Statistical coupling analysis (SCA) is a method for analyzing multiple sequence alignments that was used to identify groups of coevolving residues termed "sectors". The method applies spectral analysis to a matrix obtained by combining correlation information with sequence conservation. It has been asserted that the protein sectors identified by SCA are functionally significant, with different sectors controlling different biochemical properties of the protein. Here we reconsider the available experimental data and note that it involves almost exclusively proteins with a single sector. We show that in this case sequence conservation is the dominating factor in SCA, and can alone be used to make statistically equivalent functional predictions. Therefore, we suggest shifting the experimental focus to proteins for which SCA identifies several sectors. Correlations in protein alignments, which have been shown to be informative in a number of independent studies, would then be less dominated by sequence conservation.


Comparison of plethysmographic and helium dilution lung volumes in patients with a giant emphysematous bulla as selection criteria for endobronchial valve implant.
To assess whether the difference in lung volume measured with plethysmography and with the helium dilution technique could differentiate an open from a closed bulla in patients with a giant emphysematous bulla and could be used as a selection criterion for the positioning of an endobronchial valve. We reviewed the data of 27 consecutive patients with a giant emphysematous bulla undergoing treatment with an endobronchial valve. In addition to standard functional and radiological examinations, total lung capacity and residual volume were measured with the plethysmographic and helium dilution technique. We divided the patients into 2 groups, the collapse or the no-collapse group, depending on whether the bulla collapsed or not after the valves were put in position. We statistically evaluated the intergroup differences in lung volume and outcome. In the no-collapse group (n = 6), the baseline plethysmographic values were significantly higher than the helium dilution volumes, including total lung capacity (188 ± 14 vs 145 ± 13, P = 0.0007) and residual volume (156 ± 156 vs 115 ± 15, P = 0.001). In the collapse group, there was no significant difference in lung volumes measured with the 2 methods. A difference in total lung capacity of ≤ 13% and in residual volume of ≤ 25% measured with the 2 methods predicted the collapse of the bulla with a success rate of 83% and 84%, respectively. Only the collapse group showed significant improvement in functional data. Similar values in lung volumes measured with the 2 methods support the hypothesis that the bulla communicates with the airway (open bulla) and thus is likely to collapse when the endobronchial valve is implanted. Further studies are needed to validate our model.


Predictions derived from modelling the hippocampal role in navigation.
A computational model of the lesion and single unit data from navigation in rats is reviewed. The model uses external (visual) and internal (odometric) information from the environment to drive the firing of simulated hippocampal place cells. Constraints on the functional form of these inputs are drawn from experiments using an environment of modifiable shape. The place cell representation is used to guide navigation via the creation of a representation of goal location via Hebbian modification of synaptic strengths. The model includes consideration of the phase of firing of place cells with respect to the theta rhythm of hippocampal EEG. A series of predictions for behavioural and single-unit data in rats are derived from the input and output representations of the model.


Assessment of Device-Related Thrombus and Associated Clinical Outcomes With the WATCHMAN Left Atrial Appendage Closure Device for Embolic Protection in Patients With Atrial Fibrillation (from the PROTECT-AF Trial).
Left atrial appendage closure with the WATCHMAN device is an alternative to anticoagulation for stroke prevention in selected patients with atrial fibrillation (AF). LA device-related thrombus (DRT) is poorly defined and understood. We aimed to (1) develop consensus echocardiographic diagnostic criteria for DRT; (2) estimate the incidence of DRT; and (3) determine clinical event rates in patients with DRT. In phase 1 (training), a training manual was developed and reviewed by 3 echocardiographers with left atrial appendage closure device experience. All available transesophageal (TEE) studies in the WATCHMAN left atrial appendage system for embolic protection in patients with atrial fibrillation (PROTECT-AF) trial patients with suspected DRT were reviewed in 2 subsequent phases. In phase 2 (primary blind read), each reviewer independently scored each study for DRT, and final echo criteria were developed. Unanimously scored studies were considered adjudicated, whereas all others were reevaluated by all reviewers in phase 3 (group adjudication read). DRT was suspected in 35 of 485 patients by the site investigator, the echocardiography core laboratory, or both; 93 of the individual TEE studies were available for review. In phase 2, 3 readers agreed on 67 (72%) of time points. Based on phases 1 and 2, 5 DRT criteria were developed. In phase 3, studies without agreement in phase 2 were adjudicated using these criteria. Overall, at least 1 TEE was DRT positive in 27 (5.7%) PROTECT-AF patients. Stroke, peripheral embolism, or cardiac/unexplained death occurred in subjects with DRT at a rate of 3.4 per 100 patient-years follow-up. In conclusion, DRT were identified on at least 1 TEE in 27 PROTECT-AF patients, indicating a DRT incidence of 5.7%. Primary efficacy events in patients with DRT occurred at a rate of 3.4 per 100 patient-years follow-up, intermediate in frequency between event rates previously reported for the overall device and warfarin arms in PROTECT-AF.


Prevalence of Alternative Diagnoses and Implications for Management in Idiopathic Normal Pressure Hydrocephalus Patients.
Following Bayes theorem, ventriculomegaly and ataxia confer only a 30% chance of idiopathic Normal Pressure Hydrocephalus (NPH). When coupled with positive responses to best diagnostic testing (extended lumbar drainage), 70% of patients recommended for shunting will not actually have NPH. This is inadequate clinical care. To determine the proportion of alternative and treatable diagnoses in patients referred to a multidisciplinary NPH clinic. Patients without previously diagnosed NPH were queried from prospectively collected data. At least 1 neurosurgeon, cognitive neurologist, and neuropsychologist jointly formulated best treatment plans. Of 328 total patients, 45% had an alternative diagnosis; 11% of all patients improved with treatment of an alternative diagnosis. Of 87 patients with treatable conditions, the highest frequency of pathologies included sleep disorders, and cervical stenosis, followed by Parkinson disease. Anti-cholinergic burden was a contributor for multiple patients. Of 142 patients undergoing lumbar puncture, 71% had positive responses and referred to surgery. Compared to NPH patients, mimickers were statistically significantly older with lower Montreal Cognitive Assessment (MoCA) score and worse gait parameters. Overall, 26% of the original patients underwent shunting. Pre-post testing revealed a statistically significant improved MoCA score and gait parameters in those patients who underwent surgery with follow-up. Because the Multidisciplinary NPH Clinic selected only 26% for surgery (corroborating 30% in Bayes theorem), an overwhelming majority of patients with suspected NPH will harbor alternative diagnoses. Identification of contributing/confounding conditions will support the meticulous work-up necessary to appropriately manage patients without NPH while optimizing clinical responses to shunting in correctly diagnosed patients.


Dynamic utility: the sixth reciprocity mechanism for the evolution of cooperation.
Game theory has been extensively applied to elucidate the evolutionary mechanism of cooperative behaviour. Dilemmas in game theory are important elements that disturb the promotion of cooperation. An important question is how to escape from dilemmas. Recently, a dynamic utility function (DUF) that considers an individual's current status (wealth) and that can be applied to game theory was developed. The DUF is different from the famous five reciprocity mechanisms called Nowak's five rules. Under the DUF, cooperation is promoted by poor players in the chicken game, with no changes in the prisoner's dilemma and stag-hunt games. In this paper, by comparing the strengths of the two dilemmas, we show that the DUF is a novel reciprocity mechanism (sixth rule) that differs from Nowak's five rules. We also show the difference in dilemma relaxation between dynamic game theory and (traditional) static game theory when the DUF and one of the five rules are combined. Our results indicate that poor players unequivocally promote cooperation in any dynamic game. Unlike conventional rules that have to be brought into game settings, this sixth rule is universally (canonical form) applicable to any game because all repeated/evolutionary games are dynamic in principle.


Consolation Without Religious Topoi: Thematic Analysis of References to "Afterlife" in Secular Eulogies in Communist Czechoslovakia (1948-1989).
The article focuses on practice of secular eulogistic rhetoric in communist Czechoslovakia (1948-1989), specifically it investigates references to "afterlife" used in eulogies, which is generally conceived as allowing the bereaved to positively reappraise the death of the loved one. However, in secular eulogies, the use of "afterlife" in religious interpretation ("resurrection", "heaven") is explicitly excluded. Based on the thematic analysis of 61 secular eulogies, 2 secular interpretations of "afterlife" are identified: symbolic continuity (continuity of memory of the deceased, through his/her legacy/work done, as a part of Nature, through children/future generations, or continuity of the deceased's love) and factual (but deprived) continuity (death is presented as a sleep, journey, or pain relief).


An educational rationale for deaf students with multiple disabilities.
Deaf students with with multiple disabilities have a long history of limited opportunity, including limited access to educational opportunities available to their deaf peers. This article places the individual needs of deaf students with multiple disabilities in the context that guides much of deaf education--the importance of language acquisition. That emphasis provides a basis for placement and curriculum options for deaf students with multiple disabilities. The authors review the evolution of placement options, describe assumptions that should guide placement and curriculum decisions, and recommend practices for optimizing these students' education. Descriptions of three service delivery models--multidisciplinary, interdisciplinary, and transdisciplinary--are provided, as well as an overview of the effectiveness of person-centered planning for deaf students with multiple disabilities. Disability-specific resources are highlighted that relate to mental retardation, autism, visual impairments, learning disabilities, attention deficit hyperactivity disorder, emotional disorders, medical issues, and general resources.


Discovering misannotated lncRNAs using deep learning training dynamics.
Recent experimental evidence has shown that some long non-coding RNAs (lncRNAs) contain small open reading frames (sORFs) that are translated into functional micropeptides, suggesting that these lncRNAs are misannotated as non-coding. Current methods to detect misannotated lncRNAs rely on ribosome-profiling (Ribo-Seq) and mass-spectrometry experiments, which are cell-type dependent and expensive. Here, we propose a computational method to identify possible misannotated lncRNAs from sequence information alone. Our approach first builds deep learning models to discriminate coding and non-coding transcripts and leverages these models' training dynamics to identify misannotated lncRNAs-i.e. lncRNAs with coding potential. The set of misannotated lncRNAs we identified significantly overlap with experimentally validated ones and closely resemble coding protein sequences as evidenced by significant BLAST hits. Our analysis on a subset of misannotated lncRNA candidates also shows that some ORFs they contain yield high confidence folded structures as predicted by AlphaFold2. This methodology offers promising potential for assisting experimental efforts in characterizing the hidden proteome encoded by misannotated lncRNAs and for curating better datasets for building coding potential predictors. Source code is available at https://github.com/nabiafshan/DetectingMisannotatedLncRNAs. Supplementary data are available at Bioinformatics online.


Dental care of the pediatric cancer patient.
Although the rate for childhood cancer has remained relatively stable for the past 2 decades, there have been drastic declines in mortality due to early diagnosis and improvements in therapy. Now over 75% of children diagnosed with cancer survive more than 5 years. The pediatric dental professional plays an important role in the prevention, stabilization, and treatment of oral and dental problems that can compromise the child's health and quality of life before, during, and after the cancer treatment. This manuscript discusses recommendations for the dental care of the pediatric oncology patient.


Propeptide-mediated inhibition of cognate gingipain proteinases.
Porphyromonas gingivalis is a major pathogen associated with chronic periodontitis. The organism's cell-surface cysteine proteinases, the Arg-specific proteinases (RgpA, RgpB) and the Lys-specific proteinase (Kgp), which are known as gingipains have been implicated as major virulence factors. All three gingipain precursors contain a propeptide of around 200 amino acids in length that is removed during maturation. The aim of this study was to characterize the inhibitory potential of the Kgp and RgpB propeptides against the mature cognate enzymes. Mature Kgp was obtained from P. gingivalis mutant ECR368, which produces a recombinant Kgp with an ABM1 motif deleted from the catalytic domain (rKgp) that enables the otherwise membrane bound enzyme to dissociate from adhesins and be released. Mature RgpB was obtained from P. gingivalis HG66. Recombinant propeptides of Kgp and RgpB were produced in Escherichia coli and purified using nickel-affinity chromatography. The Kgp and RgpB propeptides displayed non-competitive inhibition kinetics with K(i) values of 2.04 µM and 12 nM, respectively. Both propeptides exhibited selectivity towards their cognate proteinase. The specificity of both propeptides was demonstrated by their inability to inhibit caspase-3, a closely related cysteine protease, and papain that also has a relatively long propeptide. Both propeptides at 100 mg/L caused a 50% reduction of P. gingivalis growth in a protein-based medium. In summary, this study demonstrates that gingipain propeptides are capable of inhibiting their mature cognate proteinases.


[The role of biological clock in glucose homeostasis].
The mechanism of the biological clock is based on a rhythmic expression of clock genes and clock-controlled genes. As a result of their transcripto-translational associations, endogenous rhythms in the synthesis of key proteins of various physiological and metabolic processes are created. The major timekeeping mechanism for these rhythms exists in the central nervous system. The master circadian clock, localized in suprachiasmatic nucleus (SCN), regulates multiple metabolic pathways, while feeding behavior and metabolite availability can in turn regulate the circadian clock. It is also suggested that in the brain there is a food entrainable oscillator (FEO) or oscillators, resulting in activation of both food anticipatory activity and hormone secretion that control digestion processes. Moreover, most cells and tissues express autonomous clocks. Maintenance of the glucose homeostasis is particularly important for the proper function of the body, as this sugar is the main source of energy for the brain, retina, erythrocytes and skeletal muscles. Thus, glucose production and utilization are synchronized in time. The hypothalamic excited orexin neurons control energy balance of organism and modulate the glucose production and utilization. Deficiency of orexin action results in narcolepsy and weight gain, whereas glucose and amino acids can affect activity of the orexin cells. Large-scale genetic studies in rodents and humans provide evidence for the involvement of disrupted clock gene expression rhythms in the pathogenesis of obesity and type 2 diabetes. In general, the current lifestyle of the developed modern societies disturbs the action of biological clock.


Exploring front-line hospital workers' contributions to patient and worker safety.
Concerns about stubbornly persistent high rates both of error-related patient injuries and of occupational injuries among healthcare workers have generated intense exploration of etiologies, interventions, and the role of underlying safety culture. Much of this work has centered on the role of physicians and nurses in health care, and suggests common issues related to safety culture. However, the role of front-line health care workers, such as nursing assistants, ward clerks, environmental service workers, food workers and transportation workers, among others, has not been explored sufficiently. This article provides the background for a workshop held in Washington, D.C., to identify gaps and opportunities for integrating front-line hospital workers into safety efforts. It provides a brief review of available information, the results of a series of focus groups of front-line workers from a single urban hospital addressing the question, and a series of framing questions for the workshop itself.


Predicted acetylcholine layer around embryonic motoneurones.
The secretion of acetylcholine (ACh) from cultures of dissociated rat purified embryonic motoneurones into the medium was measured at different times using a chemiluminescent assay. From these results it is proposed that a motoneurone is able to generate an extracellular ACh layer surrounding its membrane. This layer is expected to modulate the ACh concentration at the membrane in a range where ACh receptors can be sensitive, and thus to affect the response of the motoneurone. The influences of both membrane-bound and soluble-secreted acetylcholinesterase (AChE) were simulated and a local ACh membrane efflux was deduced. Since ACh usually exerts an inhibitory effect on neurite outgrowth, a possible autocrine influence of the ACh layer in motoneurone morphogenesis is discussed.


Liver tumor promoting effects of fenbendazole in rats.
In order to examine whether fenbendazole has tumor-promoting activity, a total of 70 male Fischer 344 rats were initiated with a single intraperitoneal injection of 100 mg/kg of diethylnitrosamine (DEN) or were given the saline vehicle alone; beginning 1 wk later, rats were given a diet containing 3,600; 1,800; 600; 200; 70; or 0 ppm of fenbendazole for 8 wk. Subgroups of 5 rats each from the DEN+ 1,800; DEN+0; 1,800; and 0 ppm groups were euthanatized after 1 wk of fenbendazole treatment, and the remaining animals were euthanatized at 8 wk. After 1 wk, relative liver weights (ratios to body weights) were significantly increased in the DEN+ 1,800 and 1,800 ppm groups, and based on light microscopy, periportal hepatocellular hypertrophy was evident in these groups. After 8 wk, relative liver weights were significantly increased in the groups given > or =600 ppm with or without DEN initiation. Periportal hepatocellular hypertrophy, characterized by a marked increase in smooth endoplasmic reticulum, was observed in the groups given > or =600 ppm with or without DEN initiation. Induction of cytochrome P-450 (CYP) 1A2, 2B1, or 4A1 was noted in the fenbendazole-treated groups with or without DEN initiation; that associated with CYP 1A2 was most marked. Positive immunostaining for anti-CYP 1A1/2 or CYP 2B1/2 was observed diffusely in the livers of animals in the DEN+1,800 and DEN+3,600 ppm groups. The numbers and areas of connexin 32 (Cx32)-positive spots per square centimeter in centrilobular hepatocytes were significantly decreased in an almost dose-dependent manner with fenbendazole treatment after DEN initiation. In situ hybridization for Cx32 mRNA revealed a remarkable decrease in its expression in the centrilobular hepatocytes in the DEN+70 ppm group. The numbers of glutathione S-transferase placental-form positive single cells (plus mini foci) were significantly increased in the DEN+ 1,800 and DEN+3,600 ppm groups. Since those agents that induce CYP 2B1/2 isozymes and reduce Cx32 in centrilobular hepatocytes have been suggested to be liver tumor promoters, the present results indicate that fenbendazole may be a liver tumor promoter.


The association between fracture rates and neighborhood characteristics in Washington, DC, children.
Effects of neighborhood contextual features have been found for many diseases, including bone fractures in adults. Our study objective was to evaluate the association between neighborhood characteristics and pediatric bone fracture rates. We hypothesized that neighborhood indices of deprivation would be associated with higher fracture rates. Pediatric bone fracture cases treated at a tertiary, academic, urban pediatric emergency department between 2003 and 2006 were mapped to census block groups using geographical information systems software. Fracture rates were calculated as fractures per 1000 children in each census block. Exploratory factor analysis of socioeconomic indicators was performed using 2000 census block data. Factor scores were used to predict odds of bone fracture at the individual level while adjusting for mean age, sex composition, and race/ethnicity composition at census block level using our sample data. We analyzed 3764 fracture visits in 3557 patients representing 349 distinct census blocks groups. Fracture rates among census blocks ranged from 0 to 207 per 1000 children/study period. Logistic regression modeling identified 2 factors (race/education and large families) associated with increased fracture risk. Census variables reflecting African American race, laborer/service industry employment, long-term block group residence, and lower education levels strongly loaded on the race/education factor. The large families factor indicated the children-to-families ratio within the block group. The poverty factor was not independently associated with fracture risk. Thus, neighborhood characteristics are associated with risk for fractures in children. These results can help inform translational efforts to develop targeted strategies for bone fracture prevention in children.


Saliva protein binding to streptococcal layers placed at different oral sites in 48 persons.
Preliminary studies of 10 subjects suggested that saliva protein binding to oral bacteria might vary among oral sites. This study investigated saliva protein binding to layers of oral streptococci in an expanded sample of 48 subjects. Those persons were at opposite extremes for unstimulated whole saliva amylase, sIgA, lactoferrin, and lysozyme in an initial screening of 128 individuals. Layers of Streptococcus gordonii Blackburn or Streptococcus oralis 10557 on enamel chips were placed on buccal left and right upper premolars and molars (UL, UR), labial upper central incisors (UC), and lingual lower central incisors (LL). After a 10-minute exposure to saliva, bacterial extracts were assayed for bound amylase, sIgA, lactoferrin, and lysozyme. Those proteins also were quantified in unstimulated whole saliva collected after chip exposure. Both strains bound significantly more amylase at UL and UR, and significantly less at UC. Blackburn bound more amylase than 10557 at all sites. Significantly less sIgA was bound at UC; strain differences for sIgA were inconsistent across sites. Significantly more lactoferrin and lysozyme were bound at LL. There were no strain differences for lactoferrin; 10557 bound significantly more lysozyme at UL and UR. Subjects at opposite extremes for saliva protein concentrations differed for bound amylase and lactoferrin; those differences were smaller than site and strain differences. Bound protein levels were correlated across sites and strains. Correlations between whole saliva and bound proteins were moderate and were most consistent at LL. These findings suggest that saliva protein effects on oral ecology may vary among oral sites.


[Various problems of prevention of ischemic heart disease].
Measures aimed at promoting health and preventing diseases form part of medical prophylaxis. Modifying the pattern of death causes has caused changes in applying approaches to implementation of preventive measures whose success largely depends on lifestyle, environment, nutrition and living conditions. It is necessary to change the population and community's attitude to health problems. There are reserves for enhancing the effectiveness of measures against coronary heart disease in medical institutions: improvement of physicians's training in prevention of diseases, active detection of patients with early CHD signs and subjects at a high risk for the disease, their timely adequate follow-up and multimodality drug and non-drug treatment.


IL-15, TIM-3 and NK cells subsets predict responsiveness to anti-CTLA-4 treatment in melanoma patients.
Despite the success of immune checkpoint blockade in melanoma, the majority of patients do not respond. We hypothesized that the T and NK cell subset frequencies and expression levels of their receptors may predict responses and clinical outcome of anti-CTLA-4 treatment. We thus characterized the NK and T cell phenotype, as well as serum levels of several cytokines in 67 melanoma patients recruited in Italy and Sweden, using samples drawn prior to and during treatment. Survival correlated with low expression of the inhibitory receptor TIM-3 on circulating T and NK cells prior to and during treatment and with the increased frequency of mature circulating NK cells (defined as CD3-CD56dim CD16+) during treatment. Survival also correlated with low levels of IL-15 in the serum. Functional experiments in vitro demonstrated that sustained exposure to IL-15 enhanced the expression of PD-1 and TIM-3 on both T and NK cells, indicating a causative link between high IL-15 levels and enhanced expression of TIM-3 on these cells. Receptor blockade of TIM-3 improved NK cell-mediated elimination of melanoma metastasis cell lines in vitro. These observations may lead to the development of novel biomarkers to predict patient response to checkpoint blockade treatment. They also suggest that induction of additional checkpoints is a possibility that needs to be considered when treating melanoma patients with IL-15.


c-Src-dependent MAPKs/AP-1 activation is involved in TNF-α-induced matrix metalloproteinase-9 expression in rat heart-derived H9c2 cells.
TNF-α plays a critical mediator in the pathogenesis of chronic heart failure contributing to cardiac remodeling and peripheral vascular disturbances. The implication of TNF-α in inflammatory responses has been shown to be mediated through up-regulation of inflammatory genes, including matrix metalloproteinase-9 (MMP-9). However, the detailed mechanisms of TNF-α-induced MMP-9 expression are largely unclear in the heart cells. Here, we demonstrated that in rat embryonic-heart derived H9c2 cells, TNF-α could induce MMP-9 mRNA expression associated with an increase in the secretion of MMP-9, determined by real-time PCR, zymography, and promoter activity assays. TNF-α-mediated responses were attenuated by pretreatment with the inhibitor of c-Src (PP1), EGFR (AG1478), PDGFR (AG1296), PI3K (LY294002), Akt (SH-5), MEK1/2 (U0126), p38 MAPK (SB202190), JNK1/2 (SP600125), or AP-1 (Tanshinone IIA) and transfection with siRNA of c-Src, EGFR, PDGFR, p110, Akt, or c-Jun. TNF-α stimulated c-Src, PDGFR, and EGFR phosphorylation, which were reduced by PP1. In addition, TNF-α-stimulated Akt phosphorylation was inhibited by PP1, AG1478, AG1296, or LY294002. We further demonstrated that TNF-α markedly stimulated p38 MAPK, p42/p44 MAPK, and JNK1/2 phosphorylation via a c-Src/EGFR, PDGFR/PI3K/Akt pathway. Finally, we showed that, in H9c2 cells, TNF-α-stimulated AP-1 promoter activity, c-Jun mRNA expression, and c-Jun phosphorylation were attenuated by PP1, AG1478, AG1296, LY294002, SB202190, SP600125, or U0126. These results suggested that TNF-α-induced MMP-9 expression is mediated through a c-Src/EGFR, PDGFR/PI3K/Akt/MAPKs/AP-1 cascade in H9c2 cells. Consequently, MMP-9 induction may contribute to cell migration and cardiovascular inflammation.


Stereoselectivity and chiral recognition in copper(I) olefin complexes with a chiral diamine
Trigonal copper(I) complexes of the chiral bidentate ligand (1S,2S)-N,N'-Bis-(mesitylmethyl)-1,2-diphenyl-1,2-ethanediamine ((S,S)-1) have been prepared with hydrocarbon olefins, as well as with allylic alcohols and ethers. The stereochemistry of the complexes has been investigated by 1H NMR spectroscopy and by combined quantum mechanics and molecular mechanics (QM/MM) computational methods. The coordinated chiral nitrogen atoms can display equal (R, R) or opposite (R, S) configuration, the latter being disfavored if steric hindrance is present above and below the coordination plane. Although the complexes exist as rapidly equilibrated mixtures of stereoisomers, one of these is often dominant, and prochiral olefins are coordinated with high enantioface selection. In addition, the [(S,S)-1]-Cu+ fragment selectively recognizes the R enantiomer of secondary allylic alcohols and ethers, as confirmed by the X-ray crystal structure analysis of the adduct with (R)-1-buten-3-ol. The reasons for the observed selectivities have been elucidated, and lead to some implications which are consistent with the enantioselection observed in catalytic cyclopropanation reactions promoted by copper complexes of the same ligand.


Bladder cancer cells secrete while normal bladder cells express but do not secrete AGR2.
Anterior gradient 2 (AGR2) is a cancer-associated secreted protein found predominantly in adenocarcinomas. Given its ubiquity in solid tumors, cancer-secreted AGR2 could be a useful biomarker in urine or blood for early detection. However, normal organs express and might also secrete AGR2, which would impact its utility as a cancer biomarker. Uniform AGR2 expression is found in the normal bladder urothelium. Little AGR2 is secreted by the urothelial cells as no measurable amounts could be detected in urine. The urinary proteomes of healthy people contain no listing for AGR2. Likewise, the blood proteomes of healthy people also contain no significant peptide counts for AGR2 suggesting little urothelial secretion into capillaries of the lamina propria. Expression of AGR2 is lost in urothelial carcinoma, with only 25% of primary tumors observed to retain AGR2 expression in a cohort of lymph node-positive cases. AGR2 is secreted by the urothelial carcinoma cells as urinary AGR2 was measured in the voided urine of 25% of the cases analyzed in a cohort of cancer vs. non-cancer patients. The fraction of AGR2-positive urine samples was consistent with the fraction of urothelial carcinoma that stained positive for AGR2. Since cancer cells secrete AGR2 while normal cells do not, its measurement in body fluids could be used to indicate tumor presence. Furthermore, AGR2 has also been found on the cell surface of cancer cells. Taken together, secretion and cell surface localization of AGR2 are characteristic of cancer, while expression of AGR2 by itself is not.


[Role of sympathectomy in upper-limb critical ischaemia induced by distal lesion of arteries].
The work was based on the experience gained in treatment of 40 patients with degree IV ischaemia of the upper extremities, induced by involvement of the distal arterial bed. Assessing blood circulation in the upper extremities included rheovasography, laser Doppler flowmetry with functional loads, and transcutaneous oxygenometry. Analyzing the obtained findings became it possible to draw a conclusion that the upper thoracic sympathectomy is an efficacious method of treatment for upper limb ischaemia secondary to the lesions of the distal arterial bed. This operative intervention turned out to be most efficient in patients with Raynaud's syndrome.


Ethical considerations for biobanking and use of genomics data in Africa: a narrative review.
Biobanking and genomic research requires collection and storage of human tissue from study participants. From participants' perspectives within the African context, this can be associated with fears and misgivings due to a myriad of factors including myths and mistrust of researchers. From the researchers angle ethical dilemmas may arise especially with consenting and sample reuse during storage. The aim of this paper was to explore these ethical considerations in the establishment and conduct of biobanking and genomic studies in Africa. We conducted a narrative synthesis following a comprehensive search of nine (9) databases and grey literature. All primary research study designs were eligible for inclusion as well as both quantitative and qualitative evidence from peer reviewed journals, spanning a maximum of 20 years (2000-2020). It focused on research work conducted in Africa, even if data was stored or analysed outside the region. Of 2,663 title and abstracts screened, 94 full texts were retrieved and reviewed for eligibility. We included 12 studies (7 qualitative; 4 quantitative and one mixed methods). Ethical issues described in these papers related to community knowledge and understanding of biobanking and genomic research, regulation, and governance of same by research ethics committees, enrolment of participants, types of informed consents, data collection, storage, usage and sharing as well as material transfer, returning results and benefit sharing. ca. Biospecimen collection and storage is given in trust and participants expect confidentially of data and results generated. Most participants are comfortable with broad consent due to trust in researchers, though a few would like to be contacted for reconsenting in future studies, and this would depend on whether the new research is for good cause. Sharing data with external partners is welcome in some contexts but some research participants did not trust foreign researchers. Biobanking and genomic studies are a real need in Africa. Linked to this are ethical considerations related to setting up and participation in biobanks as well as data storage, export, use and sharing. There is emerging or pre-existing consensus around the acceptability of broad consent as a suitable model of consent, the need for Africans to take the lead in international collaborative studies, with deliberate efforts to build capacity in local storage and analysis of samples and employ processes of sample collection and use that build trust of communities and potential study participants. Research ethics committees, researchers and communities need to work together to work together to adapt and use clearly defined ethical frameworks, guidelines, and policy documents to harmonize the establishment and running of biobanking and genomic research in Africa.


[An electrophysiological evaluation of the role of the olfactory analyzer in brain integrative activity].
Summary electrical activity of olfactory bulb and amygdala of unrested rats with reversible anosmia during conditioning with application of light and odor as conditioned stimuli was studied. It was shown, that recovery of capacity for reception of odor wasn't connected with appearance in electrical activity of investigated structures of high-frequency high-amplitude components. Additional investigations of neuronal isolation of olfactory bulb and discovery of correlative changes of power of high-frequency components in dependency on level of common excitation give the possibility to consider this rhythm as one of the mechanisms of nonsensory modulator influences of olfactory analyser.


Metastatic disease in long bones: A proposed scoring system for diagnosing impending pathologic fractures. 1989.
A weighted scoring system is proposed to quantify the risk of sustaining a pathologic fracture through a metastatic lesion in a long bone. This system objectively analyzes and combines four roentgenographic and clinical risk factors into a single score. Retrospective analysis of metastatic long bone lesions was completed in 78 lesions that had been irradiated without prophylactic surgical fixation. Clinical data and roentgenograms were scored prior to irradiation by independent observers. The outcome identified 51 lesions that did not fracture during the subsequent six months and 27 lesions that fractured within six months. A mean score of 7 was found in the nonfracture group, whereas the fracture group had a mean score of 10. The percentage risk of a lesion sustaining a pathologic fracture could be predicted for any given score. As the score increased above 7, so did the percentage risk of fracture. It is suggested that all metastatic lesions in long bones be evaluated prior to irradiation. Lesions with scores of 7 or lower can be safely irradiated without risk of fracture, while lesions with scores of 8 or higher require prophylactic internal fixation prior to irradiation.


Cardiac Structure and Function in Well-Healed Burn Survivors.
Long-term burn survivors have reduced aerobic capacity, placing them at increased risk for cardiovascular disease, morbidity, and mortality. However, the exact mechanism contributing to a reduced aerobic capacity remains incompletely understood, but may be related to adverse cardiovascular remodeling. Therefore, it was hypothesized that well-healed burn survivors would exhibit adverse left ventricular (LV) remodeling and impaired LV function. To test this hypothesis, 22 well-healed moderately burned individuals (age: 41 ± 14 years; BMI: 27.7 ± 5.4 kg/m2; male/female: 12/10; extent of burn: 37 ± 12 %BSA), 11 well-healed severely burned individuals (age: 43 ± 12 years; BMI: 29.5 ± 5.8 kg/m2; male/female: 8/3; extent of burn: 73 ± 11 %BSA), and 12 healthy, age-matched controls (age: 34 ± 9 years; BMI: 28.6 ± 5.2 kg/m2; male/female: 5/7) were enrolled in the study. All subjects were sedentary, performing less than 30 minutes of aerobic exercise per day, 3 days per week. LV morphology and function were assessed via cardiac magnetic resonance imaging. In contrast to the hypothesis, neither the presence nor severity of burn injury adversely affected LV morphology or function, when compared with equally sedentary nonburned controls. However, of note, LV mass of all three groups was in the lowest 5th percentile compared with normative values. Finally, group differences in LV morphology were largely explained by differences in aerobic capacity. Taken together, these data suggest a prior burn injury itself does not result in pathological remodeling of the LV and support a role for aerobic exercise training to improve cardiac function.


Intracanalicular schwannoma of the facial nerve: a manifestation of neurofibromatosis type 2.
Primary facial nerve tumors, which are relatively uncommon, can present a diagnostic dilemma based on their location and variable pattern of symptoms. Of primary cranial nerve tumors, schwannomas of the facial nerve rank third in frequency after those of the eighth and fifth cranial nerves. We report an illustrative case of an intracanalicular schwannoma associated with several central nervous system tumors, consistent with neurofibromatosis type 2. Initially assumed to be an eighth cranial nerve tumor, the schwannoma was found intraoperatively to arise from the facial nerve. Early diagnosis and treatment enabled excision of the tumor without sacrifice of the facial nerve. Facial nerve schwannomas can resemble acoustic schwannomas in their clinical presentation. Only a heightened level of clinical vigilance will point to the correct diagnosis and result in an optimal therapeutic outcome for patients with these rare tumors.


The utility of PET-CT in the staging and management of advanced and recurrent malignant melanoma.
Accurate pre-operative staging and correct surgical selection of patients with malignant melanoma reduces unnecessary morbidity and mortality, improves distant control and may improve survival. 18F-fluorodeoxyglucose Positron Emission Tomography Computed Tomography (18F-FDG PET-CT) has been shown to be useful in exclusion of metastatic sites and aids in surgical planning in stage III and potentially resectable stage IV disease. The primary objective of the study was to determine whether the use of PET-CT alters the initial staging and management of patients with advanced and recurrent melanoma. Retrospective analysis of clinical records of patients with malignant melanoma referred for staging PET-CT over a three-year period at our institution was performed. Pre- and post-PET-CT stage was recorded and a descriptive analysis was done to determine whether PET-CT resulted in a change in stage grouping and whether this change effected a change in clinical management. A change in stage grouping occurred in 21/39 (53.8%) of patients, 76.2% of which were up-staged and 23.8% down staged. On analysis of stage III/IV and recurrent melanoma, a change in stage occurred in 90% of stage III, 50% of stage IV and 50% of recurrent melanoma patients. This effected a change in management in 86.7% of patients with stage III, IV and recurrent melanoma collectively. PET-CT is a useful tool in the staging and subsequent management of melanoma. Its utility is pronounced in advanced and recurrent melanoma.


Relative or absolute? Implications and consequences of the measures adopted to investigate motor overflow.
Motor overflow is involuntary overt movement or covert muscle activity that cooccurs with voluntary movement. Overflow is present in several pathological conditions, as well as in neurologically healthy children and older adults, and can be induced in healthy young adults under effortful conditions. This motor phenomenon may provide insight into the underlying mechanisms and kinetic characteristics of voluntary and involuntary motor control in various populations. Although often measured behaviorally using force transduction techniques, different methods of calculating and presenting such overflow data have resulted in seemingly contradictory findings, with limited discussion of the advantages and limitations of different approaches. In this article, the authors examined the relevant literature to highlight significant methodological considerations for authors and readers conducting or appraising this type of research. Issues regarding the interpretation and reporting of findings are also discussed. Researchers are encouraged to continue using behavioral measures to create well-defined variables that enable the study of the kinematic characteristics of overflow, as these may offer promising new ways forward in better characterizing and understanding this intriguing movement phenomenon.


Gene expression profile in response to doxorubicin-rapamycin combined treatment of HER-2-overexpressing human mammary epithelial cell lines.
HER-2-positive breast cancers frequently sustain elevated AKT/mTOR signaling, which has been associated with resistance to doxorubicin treatment. Here, we investigated whether rapamycin, an mTOR inhibitor, increased the sensitivity to doxorubicin therapy in two HER-2-overexpressing cell lines: C5.2, which was derived from the parental HB4a by transfection with HER-2 and SKBR3, which exhibits HER-2 amplification. The epithelial mammary cell line HB4a was also analyzed. The combined treatment using 20 nmol/L of rapamycin and 30 nmol/L of doxorubicin arrested HB4a and C5.2 cells in S to G(2)-M, whereas SKBR3 cells showed an increase in the G(0)-G(1) phase. Rapamycin increased the sensitivity to doxorubicin in HER-2-overexpressing cells by approximately 2-fold, suggesting that the combination displayed a more effective antiproliferative action. Gene expression profiling showed that these results might reflect alterations in genes involved in canonical pathways related to purine metabolism, oxidative phosphorylation, protein ubiquitination, and mitochondrial dysfunction. A set of 122 genes modulated by the combined treatment and specifically related to HER-2 overexpression was determined by finding genes commonly regulated in both C5.2 and SKBR3 that were not affected in HB4a cells. Network analysis of this particular set showed a smaller subgroup of genes in which coexpression pattern in HB4a cells was disrupted in C5.2 and SKBR3. Altogether, our data showed a subset of genes that might be more robust than individual markers in predicting the response of HER-2-overexpressing breast cancers to doxorubicin and rapamycin combination.


Impact of guided bone regeneration and defect dimension on wound healing at chemically modified hydrophilic titanium implant surfaces: an experimental study in dogs.
The aim of the present study was to evaluate the impact of guided bone regeneration and defect dimension on wound healing at chemically modified titanium implant surfaces (modSLA). ModSLA implants were placed at chronic-type lateral ridge defects of different heights (H1-H4: 2, 4, 6 and 8 mm) and randomly allocated to either (a) GBR (polyethylene glycol membrane + biphasic calcium phosphate) or (b) untreated control. At 2 and 8 weeks (n=6 dogs each), dissected blocks were processed for histomorphometrical analysis [e.g., percentage linear fill (PLF), regenerated area (RA)]. At 8 weeks, both groups revealed comparable mean PLF (%) [ H1 (26.1 +/- 5.8)-H4 (60.4 +/- 11.8); GBR: H1 (8.3 +/- 5.3)-H4 (50.7 +/- 23.1)] and RA (mm(2)) [ H1 (2.5 +/- 0.4)-H4 (7.4 +/- 4.1); GBR: H1 (1.8 +/- 1.0)-H4 (10.8 +/- 5.9)] values. A significant difference was observed for the mean PLF values at H1 defects. It was concluded that (i) modSLA titanium implants supported bone regeneration and osseointegration at H1-H4 defects and (ii) the present GBR procedure did not seem to improve the outcome of vertical bone regeneration, but tended to increase the mean RA values.


Mutation scanning of peach floral genes.
Mutation scanning technology has been used to develop crop species with improved traits. Modifications that improve screening throughput and sensitivity would facilitate the targeted mutation breeding of crops. Technical innovations for high-resolution melting (HRM) analysis are enabling the clinic-based screening for human disease gene polymorphism. We examined the application of two HRM modifications, COLD-PCR and QMC-PCR, to the mutation scanning of genes in peach, Prunus persica. The targeted genes were the putative floral regulators PpAGAMOUS and PpTERMINAL FLOWER I. HRM analysis of PpAG and PpTFL1 coding regions in 36 peach cultivars found one polymorphic site in each gene. PpTFL1 and PpAG SNPs were used to examine approaches to increase HRM throughput. Cultivars with SNPs could be reliably detected in pools of twelve genotypes. COLD-PCR was found to increase the sensitivity of HRM analysis of pooled samples, but worked best with small amplicons. Examination of QMC-PCR demonstrated that primary PCR products for further analysis could be produced from variable levels of genomic DNA. Natural SNPs in exons of target peach genes were discovered by HRM analysis of cultivars from a southeastern US breeding program. For detecting natural or induced SNPs in larger populations, HRM efficiency can be improved by increasing sample pooling and template production through approaches such as COLD-PCR and QMC-PCR. Technical advances developed to improve clinical diagnostics can play a role in the targeted mutation breeding of crops.


Lens fluorescence and metabolic control in type 1 diabetic patients: a 14 year follow up study.
In this prospective study the authors followed the post-translational modification of the proteins of the lens of the eye by fluorometry over a period of 14 years in patients with type 1 diabetes mellitus. Twenty patients were examined at 6 years (baseline) and 20 years (follow up) after onset of diabetes mellitus. Glycaemic levels were assessed from HbA1c measured at regular intervals from the onset of diabetes mellitus. Lens fluorescence at follow up was significantly related to mean HbA1c during the study period and to lens fluorescence at baseline (p<0.0001). Sixty per cent of the variation in rate of increase in lens fluorescence during the study period was statistically attributable to glycaemia levels. The results confirm that the rate of fluorophore accumulation in the lens of adult diabetic patients is increased in proportion to glycaemic control. This parameter alone is, however, not sufficient to explain the entire variation in lens fluorophore accumulation. Consequently, lens fluorescence must be influenced by other factors before initiation and during the study period, and possibly even before onset of diabetes mellitus. The underlying mechanism could be a variation in susceptibility to lens protein denaturation by glycation.


[Restructuring of the venous bed and tissue elements of the thymus in nursing infants with pneumonia].
Studies on cadavers of 25 healthy infants and those with pneumonia have shown that the intrinsic venous bed of the thymus is structurally adapted to the construction of this organ and comprises peri- and intralobar networks, which correlates with the high thymic activity in infants, and that the thymic venous bed and tissue elements undergo consistent structural changes in pneumonia that correlate with its duration.


Application of interventional bronchoscopic therapy in eight pediatric patients with malignant airway tumors.
Malignant airway tumors in children have rarely been reported. We evaluated the safety and therapeutic effects of interventional bronchoscopic therapy in 8 children with malignant airway tumors. We retrospectively analyzed 8 children with malignant airway tumors diagnosed by pathology, and evaluated their clinical features, chest computer tomography findings and bronchoscopic manifestations. Two of the 8 pediatric patients had high-grade malignancies (lymphoma and sarcoma) and the others all had low-grade malignancies, including 2 cases with mucoepidermoid carcinoma and 4 cases with inflammatory myofibroblastic tumor. Their ages ranged from 4 to 8 years (mean, 5.7 ± 0.9). There were no specific clinical manifestations in the children, and all of them presented with various respiratory symptoms, including cough and gasping associated with hemoptysis. Chest CT indicated round intra-airway neoplasms. Obstructive pulmonary atelectasis occurred in the main bronchus of 4 patients (3 cases of the left main bronchus and 1 case of the right intermedius bronchus). All children (1 case with local anesthesia and 7 cases with general anesthesia) underwent interventional bronchoscopic therapy, including argon plasma coagulation and CO 2 cryosurgery. The success rate for the rigid procedures was 100.0% (7/7), and the cure rate after 3 months was 85.7% (6/7). A part of the tumor remained in the lung of 1 patient with inflammatory myofibroblastic tumor after bronchoscopic treatment. One patient with local anesthesia died of suffocation caused by tumor consolidation during the bronchoscopic procedure. There were no recurrences in 6 patients during the follow-up period. One recurred patient was cured at 6 months. There are no specific manifestations in children with malignant airway tumors. Interventional bronchoscopic therapy seems to be safe and effective for those tumors under general anesthesia.


Association between outer retinal alterations and microvascular changes in intermediate stage age-related macular degeneration: an optical coherence tomography angiography study.
To investigate associations between changes in retinal vessels and alterations detected by spectral domain optical coherence tomography (SD-OCT) scans in intermediate stage age-related macular degeneration (AMD). Thirty eyes of 30 patients with intermediate dry AMD were enrolled in the study. Of the cohort study, 15 eyes (changes-AMD group) showed OCT changes preceding the development of drusen-associated atrophy. A control group of healthy subjects was selected for statistical comparisons. All patients underwent an ophthalmologic evaluation, including OCT angiography (OCTA) and SD-OCT scans. Main outcome measures were superficial vessel density, deep vessel density, macular thickness. Foveal macular thickness was 215.2±32.9 μm in changes-AMD patients and was significantly thinner than no changes-AMD patients (248.3±23.3 μm, p=0.002) and healthy subjects (268.1±19.2 μm, p<0.0001). Furthermore, in the parafoveal area, the thicknesses of both the inner retina and the outer retina were reduced in the changes-AMD group, after comparison with the two other groups. Parafoveal superficial vascular plexus flow density was 43.3±2.7% in changes-AMD patients and was decreased compared with the no changes-AMD group (48.7±3.3%, p=0.003) and healthy controls (50.4±6.1%, p=0.001). A direct correlation of the superficial plexus flow density with the inner retina parafoveal macular thickness (R2=0.761, p=0.028) was found. We demonstrated an association between SD-OCT signs and retinal blood supply in patients with intermediate AMD and we showed that patients with signs predicting development of geographic atrophy have a reduced flow in superficial vascular plexus and damage of the inner and the outer retina.


Saccharomyces cerevisiae replication factor C. II. Formation and activity of complexes with the proliferating cell nuclear antigen and with DNA polymerases delta and epsilon.
Lag times in DNA synthesis by DNA polymerase delta holoenzyme were due to ATP-mediated formation of an initiation complex on the primed DNA by the polymerase with the proliferating cell nuclear antigen (PCNA) and replication factor C (RF-C). Lag time analysis showed that high affinity binding of RF-C to the primer terminus required PCNA and that this complex was recognized by the polymerase. The formation of stable complexes was investigated through their isolation by Bio-Gel A-5m filtration. A stable complex of RF-C and PCNA on primed single-stranded mp18 DNA was isolated when these factors were preincubated with the DNA and with ATP, or, less efficiently with ATP gamma S. These and additional experiments suggest that ATP binding promotes the formation of a labile complex of RF-C with PCNA at the primer terminus, whereas its hydrolysis is required to form a stable complex. Subsequently, DNA polymerase delta binds to either complex in a replication competent fashion without further energy requirement. DNA polymerase epsilon did not associate stably with RF-C and PCNA onto the DNA, but its transient participation with these cofactors into a holoenzyme-like initiation complex was inferred from its kinetic properties and replication product analysis. The kinetics of the elongation phase at 30 degrees, 110 nucleotides/s by DNA polymerase delta holoenzyme and 50 nucleotides/s by DNA polymerase epsilon holoenzyme, are in agreement with in vivo rates of replication fork movement in yeast. A model for the eukaryotic replication fork involving both DNA polymerase delta and epsilon is proposed.


Biomechanical engineering analysis of an acute papillary muscle rupture disease model using an innovative 3D-printed left heart simulator.
The severity of acute papillary muscle (PM) rupture varies according to the extent and site of the rupture. However, the haemodynamic effects of different rupture variations are still poorly understood. Using a novel ex vivo model, we sought to study acute PM rupture to improve clinical management. Using porcine mitral valves (n = 32) mounted within an ex vivo left heart simulator, PM rupture was simulated. The mitral valve was divided into quadrants for analysis according to the PM heads. Acute PM rupture was simulated by incrementally cutting from 1/3 to the total number of chordae arising from 1 PM head of interest. Haemodynamic parameters were measured. Rupture >2/3 of the chordae from 1 given PM head or regurgitation fraction >60% led to markedly deteriorated haemodynamics. Rupture at the anterolateral PM had a stronger negative effect on haemodynamics than rupture at the posteromedial PM. Rupture occurring at the anterior head of the anterolateral PM led to more marked haemodynamic instability than rupture occurring at the other PM heads. The haemodynamic effects of acute PM rupture vary considerably according to the site and extent of the rupture. Rupture of ≤2/3 of chordae from 1 PM head or rupture at the posteromedial PM lead to less marked haemodynamics effects, suggesting a higher likelihood of tolerating surgery. Rupture at the anterolateral PM, specifically the anterior head, rupture of >2/3 of chordae from 1 PM head or regurgitation fraction >60% led to marked haemodynamic instability, suggesting the potential benefit from bridging strategies prior to surgery.


The anesthetic management of children with neonatal-onset multi-system inflammatory disease.
Neonatal-onset multi-system inflammatory disease (NOMID), a rare autosomal dominantly inherited disease, belongs to a growing spectrum of autoinflammatory diseases, is characterized by urticarial rash, arthropathy, and chronic aseptic meningitis, and is associated with mutations in the cold-induced autoinflammatory gene, CIAS1, the gene that encodes the protein, cryopyrin. As little is known about the anesthetic considerations of the disease, we sought to identify the main features and respective anesthetic and perioperative implications of NOMID. We examined perianesthetic records of children with NOMID who were anesthetized for invasive diagnostic and therapeutic interventions between 2003 and 2006. In addition, we conducted an extensive literature review of the genetic, clinical, and biochemical abnormalities of the disease. Seventeen children with NOMID (median age 8 yr, range 9 mo to 11 yr) were anesthetized for diagnostic and therapeutic procedures. All patients had neurological involvement, including increased intracranial pressure, chronic aseptic meningitis, and developmental delay; 7 had bony overgrowth, 15 ocular, and 14 otological manifestations of NOMID. Despite the complexity of the disease, the perioperative course was uncomplicated, and no serious adverse events were observed. This study is the first to investigate the anesthetic implications of NOMID, an autoinflammatory disease associated with arthropathy, recurrent fevers, urticarial rash, and chronic aseptic meningitis. While for the pediatric anesthesiologist, the presence of fever and aseptic meningitis might make the conduct of anesthetics for elective procedures less desirable, our findings suggest that without evidence of active infection, even in the presence of fever and chronic aseptic meningitis, general and regional anesthesia may be conducted in patients with NOMID without untoward complications.