An analysis of trehalose, glycerol, and mannitol accumulation during heat and salt stress in a salt marsh isolate of Aureobasidium pullulans.
Polyhydroxy compounds from Aureobasidium pullulans exposed to stress treatments of heat, salt, and simultaneous heat and salt were isolated, identified, and quantified. Results from both thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC) showed that concentrations of trehalose, mannitol, and glycerol increased under stress conditions that induce osmotic- and thermotolerance in A. pullulans. The cellular concentration of trehalose increased in heat-stressed and in simultaneously heat- and salt-stressed cells but not in cells subjected to salt stress alone. Mannitol increased under all stress conditions examined, while an increase in intracellular glycerol was apparent only in salt-stressed cells. The significance of these findings in relation to stress tolerance in salt marsh environments is discussed.


Formation of core-shell-structured Zn2SnO4-carbon microspheres with superior electrochemical properties by one-pot spray pyrolysis.
Core-shell structured Zn2SnO4-carbon microspheres with different carbon contents are prepared by one-pot spray pyrolysis without any further heating process. A Zn2SnO4-carbon composite microsphere is prepared from one droplet containing Zn and Sn salts and polyvinylpyrrolidone (PVP). Melted PVP moves to the outside of the composite microsphere during the drying stage of the droplet. In addition, melting of the phase separated metal salts forms the dense core. Carbonization of the phase separated PVP forms the textured and porous thick carbon shell. The discharge capacities of the core-shell structured Zn2SnO4-carbon microspheres for the 2(nd) and 120(th) cycles at a current density of 1 A g(-1) are 864 and 770 mA h g(-1), respectively. However, the discharge capacities of the bare Zn2SnO4 microspheres prepared by the same process without PVP for the 2(nd) and 120(th) cycles are 1106 and 81 mA h g(-1), respectively. The stable and reversible discharge capacities of the Zn2SnO4-carbon composite microspheres prepared from the spray solution with 15 g PVP decrease from 894 to 528 mA h g(-1) as current density increases from 0.5 to 5 A g(-1).


The seeds from Plantago ovata lower plasma lipids by altering hepatic and bile acid metabolism in guinea pigs.
Psyllium, the husks from Plantago ovata (PO), is recognized as a potent agent in lowering plasma cholesterol. In this study, we tested the potential hypolipidemic effects of the seeds from PO and the mechanisms associated with the lowering of plasma lipids. Male Hartley guinea pigs (n = 30; 10 per group) were fed either a control diet or diets containing 7.5 or 10 g/100 g PO for 4 wk. Diets were identical in composition except for the fiber source. The control diet contained 10 g/100 g cellulose and 2.5 g/100 g guar gum, whereas the PO diets were adjusted to a total of 12.5 g/100 g fiber with cellulose. Although a dose response was not observed, plasma triglycerides and LDL cholesterol were 34 and 23% lower in the PO groups compared with the control (P < 0.01). Lecithin cholesterol acyltransferase (LCAT) and cholesterol ester transfer protein (CETP) activities were significantly affected by the PO diets. The control group had 100 and 36% higher LCAT and CETP (P < 0.01) activities, respectively, compared with the PO groups. Hepatic total and free cholesterol concentrations were not affected by PO, but cholesteryl ester concentrations were 50% (P < 0.01) lower in the PO groups compared with the control. The activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase, the rate-limiting enzyme of cholesterol synthesis was up-regulated in the PO groups by 37%. Similarly, the activity of cholesterol 7alpha-hydroxylase, the regulatory enzyme of cholesterol catabolism to bile acids was 33% higher in the PO groups (P < 0.02). Fecal bile acids were 3 times higher in the PO groups than in the control group. These results suggest that PO exerts its hypolipidemic effect by affecting bile acid absorption and altering hepatic cholesterol metabolism.


Establishment of 'normal' nervous cell lines after transfer of polyoma virus and adenovirus early genes into murine brain cells.
Brain cells from murine embryos were transfected with the polyoma virus large T or the adenovirus 5 EIA gene and, simultaneously, with the phosphotransferase coding NeoR gene. The efficiently transfected cells were selected for their resistance to Geneticin (G418) and their ability to clone at low cell density. Subsequently, most of the selected cells could be sub-cloned and continuously grown for 6-18 months so far. Their doubling time varied between 18 and 72 h. From independent transfections, more than one hundred cell lines were established. They did not exhibit a transformed phenotype, but subsequent transfection with the polyoma middle T gene induced their oncogenic transformation. The maintenance and expression of the transferred genes were verified. Most of the analyzed cell lines retained glial properties. These results suggest that the lines obtained as well as a further extension of this in vitro system should be of interest for the study of nervous cell interactions, differentiation and functions.


Gene-specific translational control of the yeast GCN4 gene by phosphorylation of eukaryotic initiation factor 2.
Phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF-2 alpha) is one of the best-characterized mechanisms for down-regulating total protein synthesis in mammalian cells in response to various stress conditions. Recent work indicates that regulation of the GCN4 gene of Saccharomyces cerevisiae by amino acid availability represents a gene-specific case of translational control by phosphorylation of eIF-2 alpha. Four short open reading frames in the leader of GCN4 mRNA (uORFs) restrict the flow of scanning ribosomes from the cap site to the GCN4 initiation codon. When amino acids are abundant, ribosomes translate the first uORF and reinitiate at one of the remaining uORFs in the leader, after which they dissociate from the mRNA. Under conditions of amino acid starvation, many ribosomes which have translated uORF1 fail to reinitiate at uORFs 2-4 and utilize the GCN4 start codon instead. Failure to reinitiate at uORFs 2-4 in starved cells results from a reduction in the GTP-bound form of eIF-2 that delivers charged initiator tRNA(iMet) to the ribosome. When the levels of eIF-2.GTP.Met-tRNA(iMet) ternary complexes are low, many ribosomes will not rebind this critical initiation factor following translation of uORF1 until after scanning past uORF4, but before reaching GCN4. Phosphorylation of eIF-2 by the protein kinase GCN2 decreases the concentration of eIF-2.GTP.Met-tRNA(iMet) complexes by inhibiting the guanine nucleotide exchange factor for eIF-2, which is the same mechanism utilized in mammalian cells to inhibit total protein synthesis by phosphorylation of eIF-2.


ZAP-70-dependent and -independent activation of Erk in Jurkat T cells. Differences in signaling induced by H2o2 and Cd3 cross-linking.
Oxidative stress in T cells induces signaling events similar to those initiated by T cell antigen receptor engagement, including tyrosine phosphorylation and activation of the critical protein-tyrosine kinase ZAP-70. Distal signaling events such as the activation of mitogen-activated protein kinases and downstream transcription factors are also initiated by oxidative stimuli. In this study P116, a ZAP-70-negative Jurkat T cell line, was used to investigate the role of ZAP-70 in mediating activation of Erk in response to H2O2. Consistent with the hypothesis that ZAP-70 is required for activation of Erk in response to an oxidative stimulus, Erk1 and Erk2 could be rapidly activated in Jurkat cells but not in P116 cells upon addition of H2O2. P116 cells became competent for H2O2-induced Erk activation upon stable transfection with wild-type ZAP-70. An in vivo ZAP-70 substrate, SLP-76, implicated in Erk activation, also became rapidly tyrosine-phosphorylated in Jurkat cells, but not in P116 cells, upon treatment with H2O2. Surprisingly, although ZAP-70 was required for H2O2-mediated Erk activation, Erk activation in response to T cell antigen receptor engagement did not require ZAP-70. In addition to demonstrating a requirement for ZAP-70 in H2O2-stimulated Erk activation, these results provide the first evidence for the existence of a ZAP-70-independent pathway for Erk activation in T cells.


Projecting the Prevalence and Costs of Metastatic Breast Cancer From 2015 through 2030.
This study projected the number of metastatic breast cancer (mBC) cases and costs (medical and productivity) attributable to mBC through 2030 among 3 age groups: younger (aged 18-44 years), midlife (aged 45-64 years), and older women (aged 65 years and older). We developed a stock/flow model in which women enter the mBC population at initial diagnosis (de novo stage IV) or through progression of an earlier-stage cancer. Women exit the mBC population through death. Input parameters by age and phase of treatment came from the US Census, Surveillance, Epidemiology, and End Results and peer-reviewed literature. In 2030, we estimated there would be 246 194 prevalent cases of mBC, an increase of 54.8% from the 2015 estimate of 158 997. We estimated total costs (medical and productivity) of mBC across all age groups and phases of care were $63.4 billion (95% sensitivity range = $59.4-$67.4 billion) in 2015 and would increase to $152.4 billion (95% sensitivity range = $111.6-$220.4 billion) in 2030, an increase of 140%. Trends in estimated costs were higher for younger and midlife women than for older women. The cost of mBC could increase substantially in the coming decade, especially among younger and midlife women. Although accounting for trends in incidence, progression, and survival, our model did not attempt to forecast structural changes such as technological innovations in breast cancer treatment and health-care delivery reforms. These findings can motivate early detection activities, direct value-driven mBC treatment, and provide a useful baseline against which to measure the effect of prevention and treatment efforts.


[Primary testicular mucinous cystadenoma: A case report and literature review].
To investigate the tissue source, clinical diagnosis, treatment and prognosis of primary testicular mucinous cystadenoma (PTMC). We retrospectively analyzed the clinical data on a case of PTMC and reviewed relevant literature. The patient underwent radical resection of the right testis after relevant preoperative examinations. Postoperative pathology indicated mucinous cystadenoma with low-grade intraepithelial neoplasia of the glandular epithelium. No recurrence was observed during an 11-month follow-up. PTMC is an extremely rare testicular and ovarian surface epithelial tumor, usually benign, rarely malignant. For the treatment of localized PTMC, radical orchiectomy is mostly recommended, while for the cases with invasion, metastasis or recurrence tendency, chemotherapy protocols for ovarian tumors can be considered.


Multiscale modeling reveals poisoning mechanisms of MgO-supported Au clusters in CO oxidation.
Catalyst deactivation mechanisms on MgO-supported Au(6) clusters are studied for the CO oxidation reaction via first-principle kinetic Monte Carlo simulations and shown to depend on support vacancies. In defect-poor MgO or in the presence of a Mg vacancy, O(2) does not bind to the clusters and the catalyst is poisoned by CO. On Au clusters interacting with O vacancies of the support, O(2) can be chemisorbed and transient activity is observed. In this case, an unexpected catalyst "breathing" mechanism (restructuring) leads to carbonate formation and catalyst deactivation, rationalizing several experimental observations. Our study underscores the importance of the cluster's charge state and dynamics on catalytic activity.


Metrifonate: overview of safety and efficacy.
Metrifonate is a cholinesterase inhibitor with a long-lasting inhibition that raises brain acetylcholine levels. It is well-absorbed and has limited binding to serum proteins. In preliminary studies of its utility in the treatment of Alzheimer disease's (AD), it led to improvements of cognition or reduced the rate of decline of cognition compared with placebo. It also benefited the global function of these patients. Side effects include nausea, cramping, and diarrhea. Metrifonate has promise as a well-tolerated treatment of the symptoms of AD.


Impact of cryopreservation and reactive oxygen species on DNA integrity, lipid peroxidation, and functional parameters in ram sperm.
Assisted reproduction using frozen-thawed semen has practical advantages, although cryopreservation is detrimental to sperm fertility in most mammals. We examined the influence of cryopreservation and reactive oxygen species (ROS) on ram sperm DNA stability (using SCSA), lipid peroxidation (LPO), chlortetracycline fluorescence (CTC) patterns, motility and viability. In Experiment 1, DNA integrity, LPO, CTC, motility and viability tests were performed on fresh and cryopreserved sperm after 0, 6, and 24 hr in synthetic oviductal fluid (SOF). In Experiment 2, fresh sperm were incubated in serum-free SOF (SOF-S; 1, 4, and 24 hr) with 0, 50, 150, or 300 microM H2O2 then assayed. Cryopreservation increased the percentage of sperm with a high DNA fragmentation index (%DFI), decreased the percentages of motile and viable sperm at thawing (0 hr), but did not affect LPO. H2O2 (150 or 300 microM) increased %DFI after 24 hr. LPO or sperm viability were not affected by H2O2, although most motility parameters decreased. H2O2 decreased the percentage of chlortetracycline pattern F sperm at 4 hr and increased the percentage of acrosome-reacted sperm (pattern AR) after 1 hr. Pooled data of Experiment 2 showed LPO was positively correlated with SCSA (r = 0.29 to r = 0.59; P < 0.05 to P < 0.01), while most motility parameters and the percentage of viable sperm were negatively correlated with LPO (r = -0.30 to r = -0.38; P < 0.05 to P < 0.01). LPO was positively correlated with the percentage of pattern AR sperm (r = 0.33; P < 0.01). Cryopreservation and H2O2 promote DNA instability in ram sperm, though motility is a more sensitive indicator of oxidative stress than the other parameters investigated.


Synthesizing 1T-1H Two-Phase Mo1-xWxS2 Monolayers by Chemical Vapor Deposition.
1T-1H metal-semiconductor interfaces in two-dimensional (2D) transition-metal dichalcogenides (TMDs) play a crucial role in utilizing the band gaps of TMDs for applications in electronic devices. Although the 1T-1H two-phase structure has been observed in exfoliated 2D nanosheets and chemically or physically treated TMDs, it cannot in principle be achieved in large-scale TMD monolayers grown by chemical vapor deposition (CVD), which is a fabrication method for electronic device applications, because of the metastable nature of the 1T phase. In this study we report CVD growth of 1T-1H two phase TMD monolayers by controlling thermal strains and alloy compositions. It was found that in-plane thermal strains arising from the difference in thermal expansion coefficients between TMD monolayers and substrates can drive the 1H to 1T transition during cooling after CVD growth. Moreover, grain boundaries in the 2D crystals act as the nucleation sites of the 1T phase and the lattice strain perturbations from alloying noticeably promote the formation of the metastable 1T phase. This work has an important implication in tailoring structure and properties of CVD grown 2D TMDs by phase engineering.


Comparative efficacy and toxicity of roxithromycin and erythromycin ethylsuccinate in the treatment of streptococcal pharyngitis in adults.
Roxithromycin is a novel oxime ether derivative of erythromycin. Previous studies have demonstrated similar in-vitro activity for roxithromycin and erythromycin. Roxithromycin has improved pharmacokinetic properties and may be a useful alternative agent for infections where erythromycin is indicated. We compared the efficacy and toxicity of roxithromycin and erythromycin ethylsuccinate in the treatment of streptococcal pharyngitis in adults. We also assessed compliance with treatment between the two agents. Roxithromycin and erythromycin ethylsuccinate were equally effective in resolving the symptoms and signs of pharyngitis. Erythromycin ethylsuccinate was superior to roxithromycin in achieving microbiological cure (90% vs. 33%, respectively). No differences were observed in toxicity or compliance with treatment between the two agents. Based on this prespective clinical trial, roxithromycin is not considered to be an acceptable alternative agent for acute streptococcal pharyngitis in adults.


Plasmid Negative Regulation of CPAF Expression Is Pgp4 Independent and Restricted to Invasive Chlamydia trachomatis Biovars.
Chlamydia trachomatis is an obligate intracellular bacterial pathogen that causes blinding trachoma and sexually transmitted disease. C. trachomatis isolates are classified into 2 biovars-lymphogranuloma venereum (LGV) and trachoma-which are distinguished biologically by their natural host cell infection tropism. LGV biovars infect macrophages and are invasive, whereas trachoma biovars infect oculo-urogenital epithelial cells and are noninvasive. The C. trachomatis plasmid is an important virulence factor in the pathogenesis of these infections. Central to its pathogenic role is the transcriptional regulatory function of the plasmid protein Pgp4, which regulates the expression of plasmid and chromosomal virulence genes. As many gene regulatory functions are post-transcriptional, we employed a comparative proteomic study of cells infected with plasmid-cured C. trachomatis serovars A and D (trachoma biovar), a L2 serovar (LGV biovar), and the L2 serovar transformed with a plasmid containing a nonsense mutation in pgp4 to more completely elucidate the effects of the plasmid on chlamydial infection biology. Our results show that the Pgp4-dependent elevations in the levels of Pgp3 and a conserved core set of chromosomally encoded proteins are remarkably similar for serovars within both C. trachomatis biovars. Conversely, we found a plasmid-dependent, Pgp4-independent, negative regulation in the expression of the chlamydial protease-like activity factor (CPAF) for the L2 serovar but not the A and D serovars. The molecular mechanism of plasmid-dependent negative regulation of CPAF expression in the LGV serovar is not understood but is likely important to understanding its macrophage infection tropism and invasive infection nature.IMPORTANCE The Chlamydia trachomatis plasmid is an important virulence factor in the pathogenesis of chlamydial infection. It is known that plasmid protein 4 (Pgp4) functions in the transcriptional regulation of the plasmid virulence protein 3 (Pgp3) and multiple chromosomal loci of unknown function. Since many gene regulatory functions can be post-transcriptional, we undertook a comparative proteomic analysis to better understand the plasmid's role in chlamydial and host protein expression. We report that Pgp4 is a potent and specific master positive regulator of a common core of plasmid and chromosomal virulence genes shared by multiple C. trachomatis serovars. Notably, we show that the plasmid is a negative regulator of the expression of the chlamydial virulence factor CPAF. The plasmid regulation of CPAF is independent of Pgp4 and restricted to a C. trachomatis macrophage-tropic strain. These findings are important because they define a previously unknown role for the plasmid in the pathophysiology of invasive chlamydial infection.


The utility of bacterial mutagenesis testing in the characterization of mobile source emissions: a review.
In summary, the work presented demonstrates that rapid, in vitro indicators of genotoxicity have been and will continue to play a valuable role in understanding the toxicity of mobile source emissions. Bacterial assays have had tremendous value in the characterization of mobile source emissions. Specifically they have had four major uses: 1) comparative screening, 2) analyzing factors that alter the genotoxicants found in emission products, 3) directing the chemical fractionation of emission organics for the identification of specific genotoxicants, and 4) analyzing the interaction of complex emission products with various mammalian systems.


[Inducible expression of non-structural protein 3 of hepatitis C virus in E. coli].
To express recombinant non-structural protein 3 of hepatitis C virus (HCV) in E. coli. The non-structural 3 (NS3) region DNA fragment of HCV was amplified by polymerase chain reaction (PCR) and inserted into inducible proeukaryotic expressive vector pET 30C(+)at Bam H1/EcoR1 sites. The competent BL21 (DE3) E.coli was transformed, and then cultured and induced with IPTG. The expressed HCV NS3 protein was confirmed with ELISA and dot blot hybridization using HCV NS3-specific single chain Fv (ScFv) antibody. 1 893 bp DNA fragment of HCV NS3 coding region was amplified by PCR technique. HCV NS3 expressive vector pET-NS3 was constructed. After transformation with pET-NS3 and induction with IPTG, recombinant HCV NS3 protein was expressed and confirmed by specific ELISA and dot blot hybridization. The recombinant HCV NS3 can be expressed in E. coli.


A structure-based proposal for a comprehensive myotoxic mechanism of phospholipase A2-like proteins from viperid snake venoms.
Envenomation via snakebites is an important public health problem in many tropical and subtropical countries that, in addition to mortality, can result in permanent sequelae as a consequence of local tissue damage, which represents a major challenge to antivenom therapy. Venom phospholipases A2 (PLA2s) and PLA2-like proteins play a leading role in the complex pathogenesis of skeletal muscle necrosis, nevertheless their precise mechanism of action is only partially understood. Recently, detailed structural information has been obtained for more than twenty different members of the PLA2-like myotoxin subfamily. In this review, we integrate the available structural, biochemical and functional data on these toxins and present a comprehensive hypothesis for their myotoxic mechanism. This process involves an allosteric transition and the participation of two independent interaction sites for docking and disruption of the target membrane, respectively, leading to a five-step mechanism of action. Furthermore, recent functional and structural studies of these toxins complexed with ligands reveal diverse neutralization mechanisms that can be classified into at least three different groups. Therefore, the data summarized here for the PLA2-like myotoxins could provide a useful molecular basis for the search for novel neutralizing strategies to improve the treatment of envenomation by viperid snakes.


5-Aminosalicylates, sulfasalazine, steroid use, and complications in patients with ulcerative colitis.
The choice between sulfasalazine and 5-aminosalicylate (5-ASA) drugs in the management of patients with ulcerative colitis often depends on idiosyncrasies of drug tolerance and control of the disease in individual patients. We sought to evaluate whether there were population differences in the effect of 5-ASA and sulfasalazine on the occurrence of clinically recognized adverse events. We also attempted to determine whether there were differences in the use of concomitant steroids and in the rates of hospitalization. We reviewed a large computerized database drawn from general practices in the United Kingdom. There we found records of 2894 patients in whom general practitioners had diagnosed ulcerative colitis, and who were receiving ongoing medical therapy specific to ulcerative colitis. The period of data availability ran from the beginning of 1990 to the latter part of 1993. The average duration of observation was 2.1 yr per patient. Patient histories were categorized into distinct periods according to the dose of 5-ASAs and sulfasalazine, steroids, and immunosuppressants, and were further separated according to the activity of ulcerative colitis. Within these categories, we examined the initiation and discontinuation of steroids, incidence of new hospitalizations for ulcerative colitis, and clinical mention of adverse events. New clinical mentions of hepatic, pancreatic, renal, and hematological events other than anemia were similar among the 5-ASAs and were very infrequent overall. Hospitalizations for ulcerative colitis occurred with similar frequency (about 15 hospitalizations per 100 patients per year) among users of those drugs. Patients receiving sulfasalazine had lower rates of initiation of prednisolone than did patients receiving 5-ASA, but sulfasalazine was used proportionately less often in patients who had been recently hospitalized, and it may be that sulfasalazine patients were somewhat less sick, overall, than were 5-ASA-using patients. The choice of drug did not affect discontinuation rates for prednisolone among established users. In the United Kingdom, during the period of this study, serious adverse reactions to drugs were not an important aspect of the management of patients with ulcerative colitis. Renal and pancreatic complications of sulfasalazine and 5-ASA therapy were extremely rare. Sulfasalazine and 5-ASA drugs have similar steroid-sparing properties. Disease-specific hospitalizations are approximately 100 times more common in ulcerative colitis patients than are serious adverse drug effects. Considerations of drug efficacy should therefore dominate the choice between therapeutic agents.


Premalignant lesions of the lower female genital tract: cervix, vagina and vulva.
Premalignant lesions of the lower female genital tract encompassing the cervix, vagina and vulva are variably common and many, but by no means all, are related to infection by human papillomavirus (HPV). In this review, pathological aspects of the various premalignant lesions are discussed, mainly concentrating on new developments. The value of ancillary studies, mainly immunohistochemical, is discussed at the appropriate points. In the cervix, the terminology and morphological features of premalignant glandular lesions is covered, as is the distinction between adenocarcinoma in situ (AIS) and early invasive adenocarcinoma, which may be very problematic. A spectrum of benign, premalignant and malignant cervical glandular lesions exhibiting gastric differentiation is emerging with lobular endocervical glandular hyperplasia (LEGH), including so-called atypical LEGH, representing a possible precursor of non HPV-related cervical adenocarcinomas exhibiting gastric differentiation; these include the cytologically bland adenoma malignum and the morphologically malignant gastric type adenocarcinoma. Stratified mucin producing intraepithelial lesion (SMILE) is a premalignant cervical lesion with morphological overlap between cervical intraepithelial neoplasia (CIN) and AIS and which is variably regarded as a form of reserve cell dysplasia or stratified AIS. It is now firmly established that there are two distinct types of vulval intraepithelial neoplasia (VIN) with a different pathogenesis, molecular events, morphological features and risk of progression to squamous carcinoma. These comprise a more common HPV-related usual type VIN (also referred to as classic, undifferentiated, basaloid, warty, Bowenoid type) and a more uncommon differentiated (simplex) type which is non-HPV related and which is sometimes associated with lichen sclerosus. The former has a relatively low risk of progression to HPV-related vulval squamous carcinoma and the latter a high risk of progression to non-HPV related vulval squamous carcinoma. Various aspects of vulval Paget's disease are also discussed.


Water-Dispersible, Responsive, and Carbonizable Hairy Microporous Polymeric Nanospheres.
Multifunctionalization of microporous polymers is highly desirable but remains a significant challenge, considering that the current microporous polymers are generally hydrophobic and nonresponsive to different environmental stimuli and difficult to be carbonized without damage of their well-defined nanomorphology. Herein, we demonstrate a facile and versatile method to fabricate water-dispersible, pH/temperature responsive and readily carbonizable hairy microporous polymeric nanospheres based on combination of the hyper-cross-linking chemistry with the surface-initiated atom transfer radical polymerization (SI-ATRP). The hyper-cross-linking creates a highly microporous core, whereas the SI-ATRP provides diverse functionalities by surface grafting of hairy functional blocks. The as-prepared materials present multifunctional properties, including sensitive response to pH/temperature, high adsorption capacity toward adsorbates from aqueous solution, and valuable transformation into well-defined microporous carbon nanospheres because of hybrid of carbonizable core and thermo-decomposable protection shell. We hope this strategy could promote the development of both functional microporous polymers and advanced hairy nanoparticles for multipurpose applications.


Processing biological images from very low light emissions.
As opposed to strong light signals (> 10(8) photons mm-2 s-1), the construction of images from object sources with low level signals (< 10(2) photons mm-2 s-1) involves a probabilistic transfer- or point-spread function. The resulting images carry considerable uncertainty or spread, restricting resolution and quantification. In this paper we propose various strategies how to reconstruct object characteristics from very low light emissions by unfolding the imaging equation. Further, calibration techniques which help to associate light emissions with the decomposition of luminogenic substrates in a spatially selective way will be discussed.


The effect of growth factors on the proliferation and differentiation of human nasal gland cells.
To elucidate a mechanism of proliferation and differentiation of nasal gland cells, we established a serum-free 3-dimensional culture system for human nasal gland (HNG) cells and examined the effects of epidermal growth factor, keratinocyte growth factor, and retinoic acid on proliferation and differentiation of cultured HNG cells. Nasal polyps were obtained from patients undergoing endoscopic endonasal sinus surgery. The HNG cells were cultured under a monolayer culture and transferred to a collagen-embedded culture using RPMI 1640 medium containing transferrin, insulin, hydrocortisone, retinoic acid, epidermal growth factor, and keratinocyte growth factor. Cell growth was measured by bromodeoxyuridine incorporation assays. To measure cell differentiation, the percentage of cells containing secretory granules, which were stained with Alcian blue in cytoplasm, was determined. In the serum-free 3-dimensional culture, the HNG cells showed ductal structures containing secretory products in a lumen. The addition of epidermal growth factor promoted the proliferation of HNG cells in its optimal concentrations, and keratinocyte growth factor also enhanced the proliferation of HNG cells. Conversely, the differentiation of HNG cells was not dependent on epidermal growth factor and keratinocyte growth factor. Retinoic acid suppressed the proliferation, but promoted the differentiation of HNG cells. Our culture system could be useful for studying the effects of various growth factors and cytokines on HNG proliferation and differentiation to better understand the mechanisms of growth and morphogenesis of nasal glands.


Neprilysin inhibitors and risk of Alzheimer's disease: A future perspective.
Alzheimer's disease (AD) is a heterogeneous neurodegenerative disease with multifaceted neuropathological disorders. AD is characterized by intracellular accumulation of phosphorylated tau proteins and extracellular deposition of amyloid beta (Aβ). Various protease enzymes, including neprilysin (NEP), are concerned with the degradation and clearance of Aβ. Indeed, a defective neuronal clearance pathway due to the dysfunction of degradation enzymes might be a possible mechanism for the accumulation of Aβ and subsequent progression of AD neuropathology. NEP is one of the most imperative metalloproteinase enzymes involved in the clearance of Aβ. This review aimed to highlight the possible role of NEP inhibitors in AD. The combination of sacubitril and valsartan which is called angiotensin receptor blocker and NEP inhibitor (ARNI) may produce beneficial and deleterious effects on AD neuropathology. NEP inhibitors might increase the risk of AD by the inhibition of Aβ clearance, and increase brain bradykinin (BK) and natriuretic peptides (NPs), which augment the pathogenesis of AD. These verdicts come from animal model studies, though they may not be applied to humans. However, clinical studies revealed promising safety findings regarding the use of ARNI. Moreover, NEP inhibition increases various neuroprotective peptides involved in inflammation, glucose homeostasis and nerve conduction. Also, NEP inhibitors may inhibit dipeptidyl peptidase 4 (DPP4) expression, ameliorating insulin and glucagon-like peptide 1 (GLP-1) levels. These findings proposed that NEP inhibitors may have a protective effect against AD development by increasing GLP-1, neuropeptide Y (NPY) and substance P, and deleterious effects by increasing brain BK. Preclinical and clinical studies are recommended in this regard.


Axillo-axillary bypass with polytetrafluoroethylene graft for coronary subclavian steal syndrome.
The purpose of this study is to report the results of axillo-axillary bypass (AAB) for coronary subclavian steal syndrome due to proximal subclavian artery occlusion. From 2003 to 2010, AAB using a polytetrafluoroethylene (PTFE) graft was performed in 11 patients with coronary subclavian steal syndrome. There was no perioperative mortality, stroke or cardiac complications. Over a mean follow-up of 36 months (range: 6-81 months), all bypass grafts have remained patent. No patient developed recurrent symptoms of myocardial ischemia. One patient died from hemorrhagic stroke at 31 months. Our results showed that AAB using a PTFE graft provides an effective and durable treatment option for coronary subclavian steal syndrome when attempted endovascular therapy of the occluded proximal subclavian artery is unsuccessful.


Local recurrence after conservative surgery and radiation therapy for ductal carcinoma in situ: Possible importance of family history.
The optimal treatment of ductal carcinoma in situ is controversial. Traditionally, women with this disease have been treated with mastectomy with excellent results, but recently the need for such extensive surgery has been questioned. Long-term data on the use of conservative surgery and radiation therapy for treatment are limited. A retrospective analysis was performed to assess treatment outcome and prognostic factors for patients with ductal carcinoma in situ treated with conservative surgery and radiotherapy. From 1976 to 1990, 76 women with ductal carcinoma in situ were treated with conservative surgery followed by radiation therapy. The median age at diagnosis was 48 years. Seventeen patients had a positive family history of breast cancer in a first-degree (n=8) or second-degree (n=9) relative. Median follow-up interval was 74 months for the 71 survivors. In 54 patients, the carcinoma was detected by mammography alone; in 13 patients, by mammography and physical examination; and in 4 patients, by physical examination with a normal mammogram; and in 5 patients, by physical examination alone without mammography. Fifty patients had re-excision after initial biopsy. Final margins were positive in 11, close in 11, negative in 34, and unknown in 20. The median volume of excised tissue was 60 cm3. The axilla was surgically staged in 30 patients (39%) and all were negative. The whole breast was irradiated to a dose of 45 to 50 Gy in all patients. Seventy-two patients also received a boost to the primary site. The median total radiation dose to the primary site was 61 Gy (range, 46 to 71). Seven patients had a recurrence in the treated breast at 16, 18, 41, 63, 72, 83, and 104 months after treatment. The 5- and 10-year actuarial rates of local recurrence were 4% and 15%, respectively. Six of seven recurrences occurred in the vicinity of the original lesion. Four local recurrences were invasive, and three were ductal carcinoma in situ. Two patients developed a contralateral invasive carcinoma. The 5- and 10-year cause-specific survival rates were 100% and 96%, respectively. The 10-year actuarial rate of local recurrence was 25% in the group with a total excision volume less than 60 cm3, as compared with 0% in those with 60 cm3 or more excised (P=0.04). In patients with a positive family history, the 10-year actuarial rate of local recurrence was 37%, as compared with 9% in patients with a negative family history (P=0.008). Of the 17 patients with a positive family history, four developed either an ipsilateral or contralateral invasive breast cancer, whereas 1 of the 58 patients without a family history developed a subsequent invasive breast cancer (P=0.008). These results suggest that patients with ductal carcinoma in situ treated with conservative surgery and radiotherapy (including a boost to the primary site) appear to benefit from wide, rather than limited, resection. These results also suggest that family history may be an important prognostic factor for progression of disease.


Tobacco cessation training among dental and other health professions students in eight low- and middle-income countries.
This study assessed differences among health professions students in exposure to didactic tobacco cessation training in asking about patients' tobacco use status ("ask") and assisting smoking patients to quit by providing educational materials ("assist"). Data from the 2005-08 Global Health Professions Student Survey were analyzed for 28,420 medical, dental, nursing, and pharmacy students in eight low- and middle-income countries. Country-specific prevalence of exposure to training in tobacco cessation was calculated for each profession category; differences were assessed using logistic regression analysis (p<0.05). The proportion of dental students taught to implement the "ask" intervention ranged from 45.4 percent (Armenia) to 95.2 percent (Chile). Only about one-third of these dental students reported being taught to implement the "assist" intervention in most countries. After adjusting for survey year, country, gender, and tobacco use, the odds of dental students' being taught to implement the "ask" intervention were lower than for medical students (adjusted odds ratio [aOR]=0.63; 95% CI: 0.42-0.96). Similarly, the odds of being taught to implement the "assist" intervention were significantly higher for medical (aOR=1.65; 95% CI: 1.26-2.17), nursing (aOR=2.84; 95% CI: 2.37-3.40), and pharmacy students (aOR=1.36; 95CI:1.05-1.76) than for dental students. These findings underscore the need for enhanced measures to incorporate tobacco cessation training as a formal component of dental education globally.


Effect of carbogen, oxygen and intraocular pressure on Heidelberg retina flowmeter parameter 'flow' measured at the papilla.
The aim of the study was to evaluate whether the Heidelberg retina flowmeter (HRF), a new device for retinal and anterior optic nerve blood flow assessment, can gauge, at least semiquantitatively, a known effect such as an increase in optic nerve blood flow by hypercapnia or a decrease in optic nerve blood flow by hyperoxia or high intraocular pressure (IOP). Measurements with the HRF were obtained at the papilla of three groups of 5 young healthy subjects (1) at baseline and after breathing 5% carbogen, (2) at baseline and after breathing 100% oxygen and (3) at baseline and after increasing IOP to 20 and 50 mm Hg. The changes in the value of the HRF parameter 'flow' were analyzed by means of a paired Student's t test. Breathing 100% oxygen for 7 min resulted in a statistically significant decrease of 34.7+/-2.5% (mean+/-SEM) in HR parameter 'flow' (p < 0.01) at the papilla. Breathing 5% carbogen for 7 min resulted in a statistically significant increase of 18.3+/-2.6% in HRF parameter 'flow' (p = 0.024). Increasing IOP to 20 mm Hg did not result in a statistically significant change in HRF parameter 'flow' (-9.6+/-7.4%; p = 0.13). Increasing IOP from 20 to 50 mm Hg, however, resulted in a statistically significant decrease of 40.1+/-6.6% in HRF parameter 'flow' (p = 0.003). With the applied stimuli, the HRF parameter 'flow' changed in the expected direction, i.e. an increase with hypercapnia and a decrease with hyperoxia or high IOP. The simplicity of use of the HRF instrument suggests that it might be well suited for a non-invasive, at least semiquantitative, assessment of changes in blood flow at the papilla.


Effective Removal of Selenite and Selenate Ions from Aqueous Solution by Barite.
In the present study, we explore a new application of Barite (BaSO4) as a sequestering phase for selenite (Se(IV)) and selenate (Se(VI)) ions from aqueous solutions because of the low solubility and high stability of Barite with its ability to selectively incorporate a large amount of various ions. The uptake of Se(IV) and Se(VI) during coprecipitation with Barite was investigated through batch experiments to understand the factors controlling effective removal of Se(IV) and Se(VI) from polluted water to Barite. The factors include (i) chemical affinity related to the degree of surface complexation between Barite surface and Se(IV)/Se(VI) ion and (ii) structural similarity related to the structural geometry of incorporated ions into the substituted site. The uptake of Se(IV) by Barite is dependent on pH, coexistent calcium ion, and sulfate concentration in the initial solution, possibly due to their effects on the chemical affinity and structural similarity. On the other hand, the uptake of Se(VI) by Barite was strongly dependent on sulfate concentration in the initial solution, which is only related to the structural similarity. This study describes the mechanisms for Se distribution between Barite and water, thereby providing a good estimate of its ability to effectively remove Se(IV) and Se(VI) from aqueous solutions under optimized experimental parameters examined here.


Coral affected by stony coral tissue loss disease can produce viable offspring.
Stony coral tissue loss disease (SCTLD) has caused high mortality of at least 25 coral species across the Caribbean, with Pseudodiploria strigosa being the second most affected species in the Mexican Caribbean. The resulting decreased abundance and colony density reduces the fertilization potential of SCTLD-susceptible species. Therefore, larval-based restoration could be of great benefit, though precautionary concerns about disease transmission may foster reluctance to implement this approach with SCTLD-susceptible species. We evaluated the performance of offspring obtained by crossing gametes of a healthy P. strigosa colony (100% apparently healthy tissue) with that of a colony affected by SCTLD (>50% tissue loss) and compared these with prior crosses between healthy parents. Fertilization and settlement were as high as prior crosses among healthy parents, and post-settlement survivorship over a year in outdoor tanks was 7.8%. After thirteen months, the diseased-parent recruits were outplanted to a degraded reef. Their survivorship was ∼44% and their growth rate was 0.365 mm ± 1.29 SD per month. This study shows that even diseased parent colonies can be effective in assisted sexual reproduction for the restoration of species affected by SCTLD.


Rictor regulates the vasculogenic mimicry of melanoma via the AKT-MMP-2/9 pathway.
Vasculogenic mimicry (VM)-positive melanomas are usually associated with poor prognosis. Rictor, the key component of the rapamycin-insensitive complex of mTOR (mTORC2), is up-regulated in several cancers, especially in melanomas with poor prognosis. The aim of this study was to investigate the role of Rictor in the regulation of VM and the mechanism underlying this possible regulation. VM channels were found in 35 of 81 tested melanoma samples and high Rictor expression correlated with VM structures. Moreover, Kaplan-Meier survival curves indicated that VM structures and high Rictor expression correlated with shorter survival in patients with melanoma. In vitro, Rictor knockdown by short hairpin RNA (shRNA) significantly inhibited the ability of A375 and MUM-2B melanoma cells to form VM structures, as evidenced by most tubes remaining open. Cell cycle analysis revealed that Rictor knockdown blocked cell growth and resulted in the accumulation of cells in G2/M phase, and cell migration and invasion were greatly affected after Rictor down-regulation. Western blotting assays indicated that down-regulating Rictor significantly inhibited the phosphorylation of AKT at Ser473 and Thr308 , which subsequently inhibited the expression and activity of downstream MMP-2/9, as confirmed by real-time PCR and gelatin Zymography. MK-2206, a small-molecule inhibitor of AKT, similarly inhibited the activity of AKT and secretion of MMP-2/9, further supporting that Rictor down-regulation inhibits the phosphorylation of AKT and activity of downstream MMP-2/9 to affect VM formation. In conclusion, Rictor plays an important role in melanoma VM via the Rictor-AKT-MMP-2/9 signalling pathway.


Dopa-responsive dystonia.
DRD is a distinctive clinical entity and an unexpectedly common subgroup of torsion dystonia. Diurnal fluctuation is often but not always present and does not reliably distinguish the disorder from ITD. DRD must be considered in the differential diagnosis of the child or adolescent presenting with a dystonic gait disorder, diplegic cerebral palsy, sporadic spastic paraplegia, ataxic syndromes, and juvenile parkinsonism. The response to L-DOPA is so dramatic and occurs so quickly that a diagnostic therapeutic trial should be undertaken in all patients presenting with these syndromes.


Interactive tele-education applied to a distant clinical microbiology specialization university course.
The microbiology laboratory provides a strategic support for infectious disease diagnosis and also alerts the medical community about bacterial resistance to antibiotics. The microbiologists' training is a challenge in Brazil, a country with an extensive territory, a diverse population, and disparity of resource allocation. The aim of this study was to implement an interactive tele-educational course in clinical microbiology to reach distant laboratory workers and to improve their professional skills. The course scientific content was defined according to competences associated, distributed in 560 h, with laboratory practices (knowledge matrix-contextual education). The 11-module course structure comprised 70% distance learning, 22% on campus (integrated modules), and 8% monographs. The group included 7 physicians and 21 microbiologists from 20 different Brazilian cities. The time flexibility and location were the two main reasons for student participation, thus decreasing absences to the workplace, different from the traditional teaching methodologies. The group performance was measured by monthly evaluations, and 1 year postcourse, the researcher visited their workplace. There was significant improvement in microbiological practices performed before compared with after group participation. Therefore, 76.9% of laboratory practices were modified because of the knowledge acquired in the course. Students showed behavioral changes in relation to performance in infection control as well as on the dissemination of their knowledge. This specialization course using distance education did not compromise the quality. This educational methodology represents an alternative to teach clinical microbiology to laboratory workers from remote hospitals, as a nationwide continuing educational strategy.


Evidence for an imino intermediate in the T4 endonuclease V reaction.
Reductive methylation and site-directed mutagenesis experiments have implicated the N-terminal alpha-amino group of T4 endonuclease V in the glycosylase and abasic lyase activities of the enzyme. NMR studies have confirmed the involvement of the N-terminal alpha-amino group in the inhibition of enzyme activity by reductive methylation. A mechanism accounting for these results predicts that a (imino) covalent enzyme-substrate intermediate is formed between the protein N-terminal alpha-amino group and C1' of the 5'-deoxyribose of the pyrimidine dimer substrate subsequent to (or concomitantly with) the glycosylase step. Experiments to verify the existence of this intermediate indicated that enzyme inhibition by cyanide was substrate-dependent, a result classically interpreted to imply an imino reaction intermediate. In addition, sodium borohydride reduction of the intermediate formed a stable dead-end enzyme-substrate product. This product was formed whether ultraviolet light-irradiated high molecular weight DNA or duplex oligonucleotides containing a defined thymine-thymine cyclobutane dimer were used as substrate. The duplex oligonucleotide substrates demonstrated a well-defined gel shift. This will facilitate high-resolution footprinting of the enzyme on the DNA substrate.


Effects of arildone on the immunogenicity of formalin-inactivated polioviruses.
Concentrated and purified Sabin and virulent strains of poliovirus types 1, 2 and 3 were inactivated with formalin at 37 C. By addition of 5.4 microM arildone, an antiviral agent, to the virus suspension, the stability of D antigen increased in both Sabin and virulent strains of all types, especially in virulent type 1 Mahoney strain. The drug had neither any inhibitory nor enhancing effect on the formalin inactivation. When antibody response was compared in guinea pigs, Sabin strains inactivated in the absence of arildone were less immunogenic against homotypic virulent strains than inactivated vaccine prepared from virulent strains. On the other hand, Sabin strains inactivated in the presence of arildone were equally immunogenic. These results indicate that it is possible to prepare from Sabin strains a potent and safe inactivated vaccine having an immunogenicity comparable to that prepared from virulent strains.


Platelet activation and signal transduction by convulxin, a C-type lectin from Crotalus durissus terrificus (tropical rattlesnake) venom via the p62/GPVI collagen receptor.
Convulxin, a powerful platelet activator, was isolated from Crotalus durissus terrificus venom, and 20 amino acid N-terminal sequences of both subunits were determined. These indicated that convulxin belongs to the heterodimeric C-type lectin family. Neither antibodies against GPIb nor echicetin had any effect on convulxin-induced platelet aggregation showing that, in contrast to other venom C-type lectins acting on platelets, GPIb is not involved in convulxin-induced platelet activation. In addition, partially reduced/denatured convulxin only affects collagen-induced platelet aggregation. The mechanism of convulxin-induced platelet activation was examined by platelet aggregation, detection of time-dependent tyrosine phosphorylation of platelet proteins, and binding studies with 125I-convulxin. Convulxin induces signal transduction in part like collagen, involving the time-dependent tyrosine phosphorylation of Fc receptor gamma chain, phospholipase Cgamma2, p72(SYK), c-Cbl, and p36-38. However, unlike collagen, pp125(FAK) and some other bands are not tyrosine-phosphorylated. Convulxin binds to a glycosylated 62-kDa membrane component in platelet lysate and to p62/GPVI immunoprecipitated by human anti-p62/GPVI antibodies. Convulxin subunits inhibit both aggregation and tyrosine phosphorylation in response to collagen. Piceatannol, a tyrosine kinase inhibitor with some specificity for p72(SYK), showed differential effects on collagen and convulxin-stimulated signaling. These results suggest that convulxin uses the p62/GPVI but not the alpha2beta1 part of the collagen signaling pathways to activate platelets. Occupation and clustering of p62/GPVI may activate Src family kinases phosphorylating Fc receptor gamma chain and, by a mechanism previously described in T- and B-cells, activate p72(SYK) that is critical for downstream activation of platelets.


Acting in solidarity: Testing an extended dual pathway model of collective action by bystander group members.
We examined predictors of collective action among bystander group members in solidarity with a disadvantaged group by extending the dual pathway model of collective action, which proposes one efficacy-based and one emotion-based path to collective action (Van Zomeren, Spears, Fischer, & Leach, 2004). Based on two proposed functions of social identity performance (Klein, Spears, & Reicher, 2007), we distinguished between the efficacy of collective action at consolidating the identity of a protest movement and its efficacy at achieving social change (political efficacy). We expected identity consolidation efficacy to positively predict collective action tendencies directly and indirectly via political efficacy. We also expected collective action tendencies to be positively predicted by moral outrage and by sympathy in response to disadvantaged outgroup's suffering. These hypotheses were supported in two surveys examining intentions to protest for Palestine in Britain (Study 1), and intentions to attend the June 4th vigil in Hong Kong to commemorate the Tiananmen massacre among a sample of Hong Kong citizens (Study 2). The contributions of these findings to research on the dual pathway model of collective action and the different functions of collective action are discussed.


Prevalence of overweight and obesity and anthropometric reference centiles for Albanian children and adolescents living in four Balkan nation-states.
Background The extent of the obesity epidemic among youth is an important public health statistic which provides an indication of the future burden of non-communicable diseases. Many developing countries, including Albania, do not have systematic and repeated monitoring systems in operation. Methods The Balkan Survey of Inactivity in Children study is a population-based cross-sectional survey including Albanian children living in the four Balkan nation-states of Albania, Kosovo, FYR Macedonia and Montenegro. In total, 19,850 children from 49 schools were approached. The LMS method was used to provide body mass index (BMI), height, weight and waist circumference reference centiles for boys and girls. The World Health Organisation (WHO) and International Obesity Task Force (IOTF) age- and sex-specific BMI cut-points were applied to evaluate adiposity levels. Results A sample of 18,460 participants aged 6-16 years old was available for creation of the BMI for age and sex reference centiles. The prevalence of overweight (including obesity) in the full sample was 21% (95% CI: 20%-21%) according to IOTF cut-off points and 28% (95% CI: 28%-29%) at WHO cut-off points. Overweight/obesity levels where higher in boys than in girls irrespective of the cut-off points applied (p<0.001). Noticeable between-country differences were observed with a prevalence of overweight/obesity of 40% (95% CI: 38%-42%) in Montenegro but only 20% (95% CI: 19%-21%) in Kosovo (WHO cut-off points). Conclusions Overweight/obesity is highly prevalent in Albanian children and adolescents. The apparent sex and country differences may inform public health actions.


Trauma to the aorta and major vessels.
Techniques for the diagnosis of blunt thoracic vascular injuries continue to evolve, and both helical CT scanning and transesophageal echocardiography are being compared with thoracic aortography. Repair of a tear in the descending thoracic aorta often is performed with the antriofemoral centrifugal pump. Patients with penetrating thoracic vascular injuries have a variety of presentations, with aortography restricted to those with proximity of wound to the superior mediastinum or the presence of a hematoma on admission chest radiograph.


Magnetic γ-Fe2O3/ZIF-7 Composite Particles and Their Application for Oily Water Treatment.
Crude oil spills are about global challenges because of their destructive effects on aquatic life and the environment. The conventional technologies for cleaning crude oil spills need to study the selective separation of pollutants. The combination of magnetic materials and porous structures has been of considerable interest in separation studies. Here, γ-Fe2O3/ZIF-7 structures were prepared by growing a ZIF-7 layer onto supermagnetic γ-Fe2O3 nanoparticles with an average size of 18 ± 0.9 nm in situ without surface modification at low temperatures. The product composite particles were characterized using X-ray diffraction, scanning electron microscopy, energy-dispersive X-ray spectroscopy, vibrating sample magnetometry, and N2 adsorption/desorption isotherms. The analyses revealed a time growth-dependent ZIF-7 rod thickness with abundant nanocavities. The γ-Fe2O3/ZIF-7 surface area available for sorption (647 m2/g) is ∼12-fold higher than that of the γ-Fe2O3 nanoparticles. Moreover, the crystal structure of γ-Fe2O3 remained essentially unchanged following ZIF-7 coating, whereas the superparamagnetism declined depending on the coating time. The γ-Fe2O3/ZIF-7 particles were highly hydrophobic and selectively and rapidly (<5 min) sorbed crude oil and other hydrocarbon pollutants from water. As high as 6 g/g of the hydrocarbon was sorbed by the γ-Fe2O3/ZIF-7 particles immersed into the hydrocarbon. A coefficient of determination, R 2 2, consistently >0.96 at all pollutant concentrations suggested a pseudo-second-order sorption kinetics. The thermal stability and 15 cycles of use and reuse confirmed a robust γ-Fe2O3/ZIF-7 sorbent.


[Preparation of the gene targeted knockout mice for human premature aging diseases, Werner syndrome, and Rothmund-Thomson syndrome caused by the mutation of DNA helicases].
The list of human RecQ helicase comprises RecQ1, BLM (Bloom syndrome), WRN (Werner syndrome), RTS (Rothmund-Thomson syndrome), and RecQ5. Of these, the defective BLM, WRN, and RTS helicases are responsible for distinct but overlapping clinical features suggesting premature aging and an enhanced risk of cancer, which apparently stems from chromosomal instability in the cells of tissues and organs where expression of the helicase genes are specified. In an effort to obtain an animal model for these diseases, we performed gene target experiments to generate the WRN and RTS knockout mice.


Altered Functional Connectivity of Cerebello-Cortical Circuit in Multiple System Atrophy (Cerebellar-Type).
Multiple system atrophy (MSA) is regarded as a progressive neurodegenerative disease mainly divided into MSA-p type with Parkinsonism and MSA-c type with cerebellar ataxia as the main symptom. However, its neural mechanism is still unclear. In this study, we only focus on the MSA-c type. The purpose of this study is to explore the functional connectivity changes of the cerebello-cortical circuit in MSA-c type by using resting state functional magnetic resonance imaging (rs-fMRI). Thirty-six subjects (18 MSA and 18 normal controls) participated in this study and the rs-fMRI data were collected by applying resting state amplitude of low-frequency fluctuations (ALFF), we found the significant decreased ALFF in the MSA patients relative to controls, which included left cerebellum 8 area, 9 area, 7b area and Cru1 as well as vermis 7. Then we select the brain region of cerebellum 8 area as seed to investigate whole brain functional connectivity alteration in the MSA patients. When comparing to controls, several regions showed decreased connectivity in the MSA patients including bilateral cerebellum anterior lobe, left cerebellum posterior lobe, left dentate, bilateral pons, inferior parietal lobule (IPL), lingual gyrus (LG), parahippocampus (PHG), and middle temporal gyrus (MTG). In addition, there were closely correlation between functional connectivities and clinical performances in the MSA patients. The current study confirmed that the disrupted functional connectivity of specific cerebello-cortical circuit in the MSA patients, which is responsible for the clinical performances.


[Rare hemoglobins in Guadeloupe and in the Lesser Antilles (author's transl)].
Screening of abnormal hemoglobins on 25 136 samples allowed us to detect from 1974-1979 rare or uncommon hemoglobins as N Baltimore, Korle Bu, Lepore, K Woolwich, J Broussais and J Baltimore. Hemoglobins N Baltimore, J Broussais, and J Baltimore are detected for the first time in Guadedoupe. The identification of these asymptomatic hemoglobins in this part of the Carribbean region provides information concerning the exact origin of the black population of these islands; e.g., hemoglobin K Woolwich and N Baltimore are found only in specific regions of West Africa.


Glutaraldehyde: behavior in aqueous solution, reaction with proteins, and application to enzyme crosslinking.
Glutaraldehyde possesses unique characteristics that render it one of the most effective protein crosslinking reagents. It can be present in at least 13 different forms depending on solution conditions such as pH, concentration, temperature, etc. Substantial literature is found concerning the use of glutaraldehyde for protein immobilization, yet there is no agreement about the main reactive species that participates in the crosslinking process because monomeric and polymeric forms are in equilibrium. Glutaraldehyde may react with proteins by several means such as aldol condensation or Michael-type addition, and we show here 8 different reactions for various aqueous forms of this reagent. As a result of these discrepancies and the unique characteristics of each enzyme, crosslinking procedures using glutaraldehyde are largely developed through empirical observation. The choice of the enzyme-glutaraldehyde ratio, as well as their final concentration, is critical because insolubilization of the enzyme must result in minimal distortion of its structure in order to retain catalytic activity. The purpose of this paper is to give an overview of glutaraldehyde as a crosslinking reagent by describing its structure and chemical properties in aqueous solution in an attempt to explain its high reactivity toward proteins, particularly as applied to the production of insoluble enzymes.


Comparative analysis of machine learning approaches to classify tumor mutation burden in lung adenocarcinoma using histopathology images.
Both histologic subtypes and tumor mutation burden (TMB) represent important biomarkers in lung cancer, with implications for patient prognosis and treatment decisions. Typically, TMB is evaluated by comprehensive genomic profiling but this requires use of finite tissue specimens and costly, time-consuming laboratory processes. Histologic subtype classification represents an established component of lung adenocarcinoma histopathology, but can be challenging and is associated with substantial inter-pathologist variability. Here we developed a deep learning system to both classify histologic patterns in lung adenocarcinoma and predict TMB status using de-identified Hematoxylin and Eosin (H&E) stained whole slide images. We first trained a convolutional neural network to map histologic features across whole slide images of lung cancer resection specimens. On evaluation using an external data source, this model achieved patch-level area under the receiver operating characteristic curve (AUC) of 0.78-0.98 across nine histologic features. We then integrated the output of this model with clinico-demographic data to develop an interpretable model for TMB classification. The resulting end-to-end system was evaluated on 172 held out cases from TCGA, achieving an AUC of 0.71 (95% CI 0.63-0.80). The benefit of using histologic features in predicting TMB is highlighted by the significant improvement this approach offers over using the clinical features alone (AUC of 0.63 [95% CI 0.53-0.72], p = 0.002). Furthermore, we found that our histologic subtype-based approach achieved performance similar to that of a weakly supervised approach (AUC of 0.72 [95% CI 0.64-0.80]). Together these results underscore that incorporating histologic patterns in biomarker prediction for lung cancer provides informative signals, and that interpretable approaches utilizing these patterns perform comparably with less interpretable, weakly supervised approaches.


Recreational windsurfing-related acute injuries: a narrative review. Part 2: injury prevention and a proposal for a set of potential prevention strategies with a holistic approach.
The purpose of this review was to identify existing prevention strategies for recreational windsurfing-related acute injuries and provide clinicians with a practical overview of current evidence supporting proposed potential prevention strategies. A literature search was conducted through March 8, 2023, using relevant keywords with Boolean operators, such as "windsurfing" AND "injury prevention" and "windsurfing" AND "exercise interventions," from the PubMed and Google Scholar databases. Only peer-reviewed English-articles were included. Existing prevention strategies, right-of-way rules, a new proposed set of eight potential primary to tertiary prevention strategies for windsurfing-related acute injuries, and proposed definitions of injury prevention levels equivalent to Haddon's matrix were identified and tabled. The proposed potential prevention strategies may facilitate clinicians in preventing recreational windsurfing-related acute injuries. Injury prevention for recreational windsurfing is under-researched. Future studies should focus on large prospective clinical trials evaluating the efficacy of prevention strategies for recreational windsurfing-related injuries.


Cytoskeletons of retinal pigment epithelial cells: interspecies differences of expression patterns indicate independence of cell function from the specific complement of cytoskeletal proteins.
In vertebrate tissue development a given cell differentiation pathway is usually associated with a pattern of expression of a specific set of cytoskeletal proteins, including different intermediate filament (IF) and junctional proteins, which is identical in diverse species. The retinal pigment epithelium (RPE) is a layer of polar cells that have very similar morphological features and practically identical functions in different vertebrate species. However, in biochemical and immunolocalization studies of the cytoskeletal proteins of these cells we have noted remarkable interspecies differences. While chicken RPE cells contain only IFs of the vimentin type and do not possess desmosomes and desmosomal proteins RPE cells of diverse amphibian (Rana ridibunda, Xenopus laevis) and mammalian (rat, guinea pig, rabbit, cow, human) species express cytokeratins 8 and 18 either as their sole IF proteins, or together with vimentin IFs as in guinea pig and a certain subpopulation of bovine RPE cells. Plakoglobin, a plaque protein common to desmosomes and the zonula adhaerens exists in RPE cells of all species, whereas desmoplakin and desmoglein have been identified only in RPE desmosomes of frogs and cows, including bovine RPE cell cultures in which cytokeratins have disappeared and vimentin IFs are the only IFs present. These challenging findings show that neither cytokeratin IFs nor desmosomes are necessary for the establishment and function of a polar epithelial cell layer and that the same basic cellular architecture can be achieved by different programs of expression of cytoskeletal proteins. The differences in the composition of the RPE cytoskeleton further indicate that, at least in this tissue, a specific program of expression of IF and desmosomal proteins is not related to the functions of the RPE cell, which are very similar in the various species.


Biases in the perceptions of the consequences of alcohol use among college students.
Previous studies have shown that college students' perceptions of the quantity and frequency of peer alcohol consumption are biased. Most students report that their social referents drink more than they themselves do. In the current study members of two fraternities and two sororities (N = 252) were asked to make two types of ratings of alcohol-related consequences across four target individuals. The ratings were: (1) the frequency of occurrence of predefined alcohol-related negative consequences and (2) the degree to which certain alcohol-related consequences are considered "problems." The targets were: themselves, their best friend, a typical member of their fraternity or sorority and a typical student at the university. Estimates of frequency of problem behaviors for typical member of the residence and typical student at the university were significantly higher than ratings for self (p < .001). Data from ratings of behavioral definitions of alcohol problems for the same four targets replicated the bias, although to a lesser degree. Correlational analyses suggest that biased perceptions of problems were unrelated to personal levels of alcohol consumption. Results are discussed in terms of the cognitive and motivational factors that could result in this misrepresentation of peer behavior.


Care Practices of Mental Health Clinical Pharmacist Practitioners Within an Interdisciplinary Primary Care Model for Patients With Substance Use Disorders.
Clinical pharmacist practitioners (CPPs) play an increasingly important role in interdisciplinary care for patients with substance use disorders (SUDs). However, CPPs' scope of practice varies substantially across clinics and settings. We sought to describe CPP practices and activities within an interdisciplinary, team-based primary care clinic dedicated to treat Veterans with histories of substance use disorders, experience of homelessness, high medical complexity, and other vulnerabilities. We conducted a retrospective cohort study of CPP activities using Department of Veterans Affairs (VA) administrative data in 2019. CPPs provided care for 228 patients, including 766 in-clinic visits, 341 telephone visits, and 626 chart reviews, with an average of 2.5 hours spent per patient per year. Patients seen by CPPs frequently experience mental health conditions and SUDs, including depression (66%), post-traumatic stress disorder (52%), opioid use disorder (OUD) (45%), and alcohol use disorder (44%). CPPs managed buprenorphine medications for OUD or chronic pain in 76 patients (33%). Most CPP interventions (3330 total) were for SUDs (33%), mental health conditions (24%), and pain management (24%), with SUD interventions including medication initiation, dose changes, discontinuations and monitoring. As part of opioid risk mitigation efforts, CPPs queried the state's prescription drug monitoring program 769 times and ordered 59 naloxone kits and 661 lab panels for empaneled patients. CPPs managed a high volume of vulnerable patients and provided complex care within an interdisciplinary primary care team. Similar CPP roles could be implemented in other primary care settings to increase access to SUD treatment.


Development of the Bowman-Birk inhibitor for oral cancer chemoprevention and analysis of Neu immunohistochemical staining intensity with Bowman-Birk inhibitor concentrate treatment.
Cancer chemoprevention is a rapidly evolving approach to reverse or inhibit carcinogenesis, and there is active interest in development of effective chemopreventive agents against head and neck cancers. The retinoids are archetypal chemopreventive agents for oral premalignant lesions. They have significant clinical effect, but widespread use is limited by significant clinical toxicity. The Bowman-Birk Inhibitor is one of several nontoxic compounds exhibiting both potent anticarcinogenic activity and minimal toxicity. The purposes of the study were to summarize the preclinical and clinical development of Bowman-Birk Inhibitor and a Bowman-Birk Inhibitor concentrate against oral premalignant lesions and to evaluate Neu immunohistochemical staining intensity for lesions and simultaneously obtained biopsy specimens of normal-appearing mucosa from the Phase IIa Bowman-Birk Inhibitor concentrate oral leukoplakia chemoprevention trial. Part I is a selected literature review. Part II is a retrospective analysis of pathological specimens prospectively obtained from the Phase IIa clinical trial of Bowman-Birk Inhibitor concentrate. Thirty-two sets of biopsy specimens from lesions and uninvolved oral mucosa before and after treatment with Bowman-Birk Inhibitor concentrate in doses ranging from 200 to 1066 chymotrypsin inhibitory units were examined in blinded fashion for Neu immunohistochemical staining intensity using the 3B-5 monoclonal antibody. Staining intensity scores among the lesion and control biopsy specimens before and after Bowman-Birk Inhibitor concentrate treatment were analyzed and compared with previously obtained values for serum Neu, oral mucosal cell Neu, protease activity, and clinical response to treatment. Mean Neu staining score was significantly higher in lesions compared with uninvolved mucosa (P <.001). Pretreatment staining scores for biopsy specimens of lesions and control biopsy specimens of normal-appearing tissues were correlated (Spearman correlation coefficient [r] = 0.375, P =.045), but no correlation between lesion and control biopsy specimen scores was evident after treatment. The change in Neu staining score with Bowman-Birk Inhibitor concentrate treatment in control site biopsy specimens demonstrated an inverse relationship of change in lesion area with Bowman-Birk Inhibitor concentrate treatment (Spearman r = -0.493, P <.007). Bowman-Birk Inhibitor concentrate shows promise to become an effective nontoxic chemopreventive agent based on results of extensive preclinical studies, and Phase I and Phase IIa clinical trials. Bowman-Birk Inhibitor concentrate has dose-related clinical activity against oral leukoplakia and modulates levels of Neu and protease activity. The current investigation identified increased Neu staining intensity in hyperplastic lesions compared with simultaneously obtained biopsy specimens of normal-appearing mucosa both before and after Bowman-Birk Inhibitor concentrate treatment. This finding supports prior observations that increased Neu expression is present in a subset of oral premalignant lesions and head and neck cancers. The trend of increased Neu staining score in control biopsy tissues of subjects exhibiting decreased lesion area following Bowman-Birk Inhibitor concentrate treatment raises questions about the mechanisms of Bowman-Birk Inhibitor concentrate action. One possible explanation is that Bowman-Birk Inhibitor stabilizes the extracellular domain of Neu, thereby preventing receptor truncation and internalization. Further study of modulation of Neu and protease activity by Bowman-Birk Inhibitor concentrate treatment may provide insights into the role of proteases and protease inhibitors in oral premalignant lesions and the mechanisms underlying Bowman-Birk Inhibitor concentrate effects. A Phase IIb randomized, placebo-controlled clinical trial to determine the clinical effectiveness of Bowman-Birk Inhibitor concentrate and further evaluate these candidate biomarkers is under way.


Determinants of the clinical expression of amoxicillin-clavulanate hepatotoxicity: a prospective series from Spain.
Amoxicillin-clavulanate (AC) hepatotoxicity has been reported to exhibit a higher predominance of cholestatic types of damage, especially in males. However, the determinants of its clinical expression are unknown. This study prospectively evaluated the profile of AC hepatotoxicity. Data on all cases of hepatotoxicity reported to the Spanish Registry attributed to AC and assessed as definite or probable on the Council for International Organizations of Medical Sciences (CIOMS) scale were collated and compared to published case series. Hepatotoxicity related to amoxicillin-clavulanate was identified in 69 patients (36 males; mean age 56 years) representing 14% of all cases of hepatotoxicity submitted to the Registry. There was an overall sex distribution and the predominant pattern of lesion was hepatocellular (36%) which occurred at a shorter duration of treatment (P < .03). Mean time lapse between therapy initiation and jaundice onset was 16 days. Late onset of symptoms following end of treatment occurred in half the cases. Multiple logistic regression analysis identified advancing age as the factor associated with the development of cholestatic/mixed type of injury (odds ratio for an age interval for 1 year: 1.045 [95% CI = 1.013-1.078; P = .005). An unfavorable outcome was seen in 7% of patients. In conclusion, age is the most important determinant in the biochemical expression of AC hepatotoxicity; younger age is associated with cytolytic damage and shorter treatment duration, whereas cholestatic/mixed type of damage is related to older age and prolonged AC therapy.


Serum thyroglobulin, a monitor of differentiated thyroid carcinoma in patients receiving thyroid hormone suppression therapy: concise communication.
Serum thyroglobulin levels were obtained in 86 patients who had undergone thyroidectomy and I-131 ablation for differentiated thyroid cancer, and who were receiving or had recently discontinued thyroid hormone suppression therapy. Excellent correlation was observed between serum thyroglobulin levels in patients receiving thyroid hormone suppression therapy and I-131 imaging studies. Serum thyroglobulin levels equal to or below 20 ng/ml indicate the absence of thyroid carcinoma, and values exceeding 60 ng/ml were indicative of active thyroid cancer but may include some patients without clinical evidence of disease. Intermediate serum thyroglobulin levels were observed in a small number of patients with postsurgical thyroid remnants or active disease. Serum thyroglobulin levels are of considerable value in monitoring the activity of thyroid cancer in patients who are receiving thyroid hormone suppression therapy.


Coming to America: betel nut and oral submucous fibrosis.
Oral submucous fibrosis (OSF) is an insidious precancerous disease affecting the oral cavity, pharynx and upper digestive tract. Its etiology is linked directly to betel nut use, which is common to the Indian subcontinent. With the increase in immigration of people from the Indian subcontinent to the United States, many American dental professionals will encounter this disease in the near future. The author provides a general overview of OSF. The author provides a literature review of OSF, including its epidemiology, etiology, clinical presentation, histopathology and treatment modalities. OSF is a precancerous oral disease linked definitively to the use of betel nut. It is endemic to people in the Indian subcontinent. Patients' primary complaint is progressive trismus. Treatment is based on disease severity. OSF is a debilitating but preventable oral disease. Its incidence in the United States will increase as the South Asian immigrant population expands. Consequently, American dental professionals may encounter patients with this disease, and it is important that they are aware of it. In addition, for dental care professionals practicing in a South Asian American community, public health education is important to inform the population about the deleterious oral health effects of betel nut consumption.


A Case of Tumor-Induced Osteomalacia Detected by Venous Sampling.
Tumor-induced osteomalacia (TIO) can cause osteomalacia due to excessive production of fibroblast growth factor 23 (FGF23) by the tumor. Since TIO is a very rare disease, it is often misdiagnosed as intervertebral disc herniation, spondyloarthritis, or osteoporosis. We report a 65-year-old man who developed generalized arthralgia and difficulty walking two years ago and was diagnosed with multiple fractures throughout his body. He was initially diagnosed with osteoporosis and was treated with calcitriol. However, he was referred to our hospital since his symptoms did not improve. We diagnosed tumor-induced osteomalacia based on low serum phosphorus, high bone-type alkaline phosphatase, high FGF23 levels, and the presence of two tumors. The responsible tumor was identified using FGF23 levels in venous sampling. As the location of the tumor made surgical resection difficult, we selected treatment with burosumab, a human monoclonal antibody against FGF23, leading to improvement in the hypophosphatemia and pain, such that he was able to walk with a cane. In cases of osteoporosis with hypophosphatemia, general physicians should keep TIO in mind, and attempt to identify the responsible tumor lesion.


Pest status, bio-ecology and management of the false codling moth, Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) and its implication for international trade.
The false codling moth (FCM), Thaumatotibia leucotreta (Lepidoptera: Tortricidae) is an insect pest which represents an important threat to the production and marketing of a wide range of agricultural crops in the African-Caribbean-Pacific (ACP) countries. The FCM reduces not only the yield and quality of the crop but also as a quarantine insect pest, restricts the trade of susceptible agricultural produce on the international market. In addition, little research has been conducted in the ACP countries on the bio-ecology and sustainable management of this pest, especially on vegetables for export. Thus, action-oriented research aimed at understanding the bio-ecology of this important pest is essential to achieve effective management. Various management interventions against this pest have been used in some parts of the world, especially in South Africa on citrus. Currently, farm sanitation is regarded as the key management strategy. Exploring and improving on other interventions such as Sterile Insect Technique, monitoring and mass trapping of male moths, augmentative biological control, use of bio-pesticides, protected cultivation and cold treatment may help to mitigate the expansion of FCM into other countries, especially in the European and Mediterranean Plant Protection Organization region where it has become a regulated insect pest since 2014. This review discussed the bio-ecology of FCM and highlighted some of the challenges and opportunities for its effective management and its implication for international trade, especially the export of chillies from the ACP countries into the European Union market which requires strict phytosanitary regulations.


Purification and properties of laminaribiose phosphorylase (EC 2.4 1.31) from Euglena gracilis Z.
Three isoforms of laminaribiose phosphorylase, F0, F1, and F2, were purified to an electrophoretically homogeneous state from a cell free extract of Euglena gracilis Z (IAM E-6). F1 and F2 were the major components. The three enzymes showed very similar properties except their isoelectric points. They could also use laminaribiose as the glucosyl acceptor instead of glucose. The isoforms the same molecular mass of 120 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) or 200 kDa by the gel filtration method, suggesting that they have a dimer structure. They could not be distinguished on the Ouchterlony's double diffusion test with mouse antiserum against F1 or F2. The substrate specificities of F1 and F2 were determined to be essentially the same. Both of the reaction mechanisms of F1 and F2 were determined to be ordered bi-bi mechanisms, as in the case of cellobiose phosphorylase. A competitive substrate inhibition was observed in their synthetic reaction. Two other strains, E. gracilis var. bacillaris (IAM E-2) and E. gracilis (IAM E-3), had only one laminaribiose phosphorylase, which corresponded to F0 on native PAGE.


Advances in osteoclast biology reveal potential new drug targets and new roles for osteoclasts.
Osteoclasts are multinucleated myeloid lineage cells formed in response to macrophage colony-stimulating factor (M-CSF) and receptor activator of NF-κB ligand (RANKL) by fusion of bone marrow-derived precursors that circulate in the blood and are attracted to sites of bone resorption in response to factors, such as sphingosine-1 phosphate signaling. Major advances in understanding of the molecular mechanisms regulating osteoclast functions have been made in the past 20 years, mainly from mouse and human genetic studies. These have revealed that osteoclasts express and respond to proinflammatory and anti-inflammatory cytokines. Some of these cytokines activate NF-κB and nuclear factor of activated T cells, cytoplasmic 1 (NFATc1) signaling to induce osteoclast formation and activity and also regulate communication with neighboring cells through signaling proteins, including ephrins and semaphorins. Osteoclasts also positively and negatively regulate immune responses and osteoblastic bone formation. These advances have led to development of new inhibitors of bone resorption that are in clinical use or in clinical trials; and more should follow, based on these advances. This article reviews current understanding of how bone resorption is regulated both positively and negatively in normal and pathologic states.


Identification of T-cell-activating recombinant antigens shared among three candidate antileprosy vaccines, killed M. leprae, M. bovis BCG, and mycobacterium w.
Antigenic crossreactivity among three candidate antileprosy vaccines, killed Mycobacterium leprae, BCG, and Mycobacterium w, was studied using T-cell lines and clones raised from BCG- and killed-M. leprae-vaccinated subjects. To identify the crossreactive antigens, the T-cell lines and clones were tested against Escherichia coli lysates containing 65-, 36-, 28-, 18-, and 14-kilodalton (kDa) and 13B3 M. leprae antigens and 65-, 19-, and 12-kDa M. tuberculosis antigens. The short-term T-cell lines, which compared to T-cell clones are easy to raise and maintain, were equally effective in identifying the T-cell-activating recombinant antigens. The reactivity pattern of the T-cell lines and the clones suggested that 65-kDa M. leprae and M. tuberculosis antigens are present in M. leprae, BCG, and Mycobacterium w; 18-kDa M. leprae antigen is shared between M. leprae and Mycobacterium w, 13B3 M. leprae antigen is possessed by M. leprae and BCG. These and other unidentified T-cell-activating antigens shared among candidate leprosy vaccines may be the basis for induction of in vivo sensitization to M. leprae antigens after vaccination with BCG or Mycobacterium w.


Patterning of the cardiac outflow region in Drosophila.
Specification of bilateral cardiac primordia and formation of the linear heart tube are highly conserved from Drosophila to humans. However, subsequent heart morphogenesis involving nonmesodermal neural crest cells was thought to be specific for vertebrates. Here, we provide evidence that a group of nonmesodermal cells that we have named heart-anchoring cells (HANCs) contribute to heart morphogenesis in Drosophila. We show that the homeobox genes ladybird (lb) known to be involved in diversification of cardiac precursors are expressed in HANCs and required for their specification. Interestingly, the HANCs selectively contact the anterior cardiac cells, which express lb as well. Direct interaction between HANCs and cardiac cells is assisted by a pair of cardiac outflow muscles (COMs), each of which selectively attaches to both the lb-expressing cardiac cells and HANCs. COM muscles seem to ensure ventral bending of the heart tip and together with HANCs determine the spatial positioning of the cardiac outflow region. Experimentally depleted cardiac lb expression leads to the disruption of the contact between the tip of the heart and either the COM muscles or the HANC cells, indicating a pivotal morphogenetic role for the lb expression within the heart.


Evaluation of KRAS, NRAS and BRAF hotspot mutations detection for patients with metastatic colorectal cancer using direct DNA pipetting in a fully-automated platform and Next-Generation Sequencing for laboratory workflow optimisation.
Assessment of KRAS, NRAS (RAS) and BRAF mutations is a standard in the management of patients with metastatic colorectal cancer (mCRC). Mutations could be assessed using next-generation sequencing (NGS) or real-time PCR-based assays. Times to results are 1 to 2 weeks for NGS and 1 to 3 days for real-time PCR-based assays. Using NGS can delay first-line treatment commencement and using PCR-based assays is limited by the number of possible analysed targets. The Idylla system is a real-time PCR cartridge-based assay, able to analyse hotspots mutations using one section of FFPE tumour tissue sample. To combine short delays and analysis of a large gene-panel, we propose here a laboratory workflow combining the Idylla system and NGS and compatible with FFPE samples with low tissue quantity. In this study we evaluated and validated the Idylla system for the analysis of RAS and BRAF mutations by pipetting directly DNA in the cartridge instead of FFPE section as recommended by the manufacturer. DNA extracted from 29 FFPE samples from mCRC patients with NGS-characterized RAS and BRAF mutations were tested with the Idylla KRAS and the Idylla NRAS-BRAF mutation tests to assess sensitivity, specificity, reproducibility and limit of detection of each test. A 100% concordance was found between NGS and Idylla results for the determination of KRAS (12/12), NRAS (12/12) and BRAF (11/11) mutations with a sensitivity and a specificity of 100%. The system showed a good reproducibility with CV inferior to 3%. LOD was reached with 2.5 ng of DNA for KRAS and NRAS mutations and 5 ng of DNA for BRAF mutations. The analysis of RAS and BRAF mutations using DNA pipetted directly in the cartridge of the Idylla system showed a good sensitivity, specificity, reproducibility and LOD, and can be integrated in a laboratory workflow for samples with few tissue without compromising a further complete tumour characterization using NGS.


Analysis of the incidence of tumor lysis syndrome in patients with hematological malignancies treated with rasburicase.
The purpose of this study was to assess the incidence of tumor lysis syndrome (TLS) in patients with hematological malignancies treated with rasburicase, and to evaluate the dose and duration of rasburicase administration. A total of 52 patients were enrolled. The background of the patients, incidence of TLS and laboratory data were retrospectively examined; in addition, the dose and duration of rasburicase administration and the factors affecting the onset of TLS were evaluated and compared among TLS risk categories. During the study period, 2 (3.8%) of the patients developed clinical TLS and 24 (46.2%) developed laboratory TLS (LTLS). Although the LTLS rate was very high, there were no life-threatening cases of TLS. The median daily dose of rasburicase administered to all patients was 7.5 mg/day (interquartile range, 7.5-9.0 mg/day), and the daily weight-based dose was 0.147 mg/kg/day (range, 0.126-0.178 mg/kg/day). The administration duration was 3 days (interquartile range, 3-4 days). Additionally, there was no significant association between TLS risk classification and daily rasburicase administration dose, duration, or post-administration laboratory data. The factors affecting the onset of TLS included serum uric acid level, as well as serum creatinine and phosphate levels. Rasburicase was highly effective in the prevention and management of hyperuricemia, even at a low-dose (7.5 mg/day) and a duration that was 3 days shorter compared with that recommended by the manufacturer. Therefore, clinicians should administer rasburicase based on their clinical judgment, taking into consideration the cost-effectiveness of this therapy.


[A clinical evaluation of distant metastases from a hepatocellular carcinoma with reference to location, frequency, symptoms, treatment & prognosis].
A clinical analysis of patients with hepatocellular carcinoma (HCC) associated with a distant metastases has been undertaken by the authors. Studied were eight patients (Group A) whose distant metastases were detected at the time of their initial examination, and 24 patients (Group B) whose distant metastases had been diagnosed during the course of treatment of their primary lesions. Complications arising from liver cirrhosis in Group A was less than in Group B. The location of the metastases were the bone in 16, the lung and the adrenal gland in four, the brain and the skin in two and the kidney in one. The mean survival times were 4.6 months (Group A), and 6.3 months (Group B). The cause of death was chiefly a progression of the primary lesion, except for one brain metastases. This analysis had shown that treatment of a bone metastasis is necessary for the relief of pain, and that care should be taken for the prevention of a fracture or paralysis, and that control of the primary lesion is necessary for improving the chances of survival.


Use of Patient and Investigator Global Impression Scales: A Review of Food and Drug Administration-Approved Labeling, 2009 to 2019.
Identify disease categories in which single-item global impression (GI) scales were included in product labeling of new drugs approved by the US Food and Drug Administration (FDA) in January 2009-December 2019 and review the characteristics of GIs included in product labeling of new FDA-approved drugs (January 2017-December 2019). FDA Clinical Outcome Assessment (COA) Compendium was reviewed for drug labels that included GIs for drugs approved in 2009-2016. The indication, year of approval, ICD-10 code, and GI respondent were noted. A manual review of labels of FDA-approved drugs (2017-2019) was undertaken to identify GIs included in the labels. Corresponding drug approval packages were reviewed to identify details of any regulatory reviewer comments related to GIs. GI characteristics were noted from the drug label or the review documents, including the respondent, type of measure (static or dynamic), item wording, concept assessed, and response options. Product labeling containing GIs was most common in diseases related to the skin, nervous system, behavioral disorders, and the musculoskeletal system. GIs were included in 30/77 (39.0%) drug labels in the four disease categories. In the past 10 years, GIs have been included as endpoint measures in confirmatory clinical trials and have generated evidence of treatment benefit in diseases related to the skin, nervous system, behavioral disorders, and the musculoskeletal system. GIs frequently provide important insights into the patient experience. Before GIs are included in clinical trials to assess treatment benefit, it is important to ensure that they are valid, reliable, and responsive.


Time course and reversibility of ethanol's suppressive effects on axon sprouting in the dentate gyrus of the adult rat.
Ethanol was administered chronically to adult rats in a liquid diet for 14 days preceding and for 5, 7, 8, 9, or 10 days following the unilateral destruction of the entorhinal cortex. Control groups received a diet of lab chow and water and were sacrificed at comparable survival times. An additional experimental group was given ethanol until 9 days after the lesion, then switched to lab chow and water and sacrificed 1 day later. Coronal sections through the dorsal hippocampal formation were stained and analyzed histochemically for the localization of acetylcholinesterase (AChE). Quantitative measurements of the histochemical patterns in the molecular layer of the dentate gyrus were obtained. Ethanol exposure inhibited the withdrawal of the acetylcholinesterase-stained septohippocampal fibers and limited the typical lesion-induced expansion of the pale-staining commissural/associational zone in the molecular layer of the denervated dentate gyrus. However, abstinence from ethanol for just 24 h released the inhibitory effect on the acetylcholinesterase-staining fibers, resulting in a significant expansion of the commissural/associational zone.


Neuromyopathy with congenital cataracts and glaucoma: a distinct syndrome caused by POLG variants.
We identified three non-related patients manifesting a childhood-onset progressive neuromyopathy with congenital cataracts, delayed walking, distal weakness and wasting, glaucoma and swallowing difficulties. Electrophysiology and nerve biopsies showed a mixed axonal and demyelinating neuropathy, while muscle biopsy disclosed both neurogenic and myopathic changes with ragged red fibers, and muscle MRI showed consistent features across patients, with a peculiar concentric disto-proximal gradient of fatty replacement. We used targeted next generation sequencing and candidate gene approach to study these families. Compound biallelic heterozygous variants, p.[(Pro648Arg)]; [(His932Tyr)] and p.[(Thr251Ile),(Pro587Leu)]; [(Arg943Cys)], were found in the three patients causing this homogeneous phenotype. Our report on a subset of unrelated patients, that showed a distinct autosomal recessive childhood-onset neuromyopathy with congenital cataracts and glaucoma, expands the clinical spectrum of POLG-related disorders. It also confirms the association between cataracts and neuropathy with variants in POLG. Early onset cataract is otherwise rare in POLG-related disorders and so far reported only in a few patients with the clinical pattern of distal myopathy or neuromyopathy.