Gliosis Can Impede Integration Following Photoreceptor Transplantation into the Diseased Retina.
Retinal degenerations leading to the loss of photoreceptor (PR) cells are a major cause of vision impairment and untreatable blindness. There are few clinical treatments and none can reverse the loss of vision. With the rapid advances in stem cell biology and techniques in cell transplantation, PR replacement by transplantation represents a broad treatment strategy applicable to many types of degeneration. The number of donor cells that integrate into the recipient retina determines transplantation success, yet the degenerating retinae presents a number of barriers that can impede effective integration. Here, we briefly review recent advances in the field of PR transplantation. We then describe how different aspects of gliosis may impact on cell integration efficiency.


Calcium and chemotropic turning of nerve growth cones.
Cultured Xenopus spinal neurons exhibit chemotropic turning toward the source of neurotransmitters acetylcholine and glutamate. Here we review the experimental evidence that transmitter-induced turning of the growth cone is mediated by an influx of Ca2+, that a gradient of intracellular Ca2+ within the growth cone is responsible for the directional growth cone response, and that asymmetric filopodia formation precedes and is essential for the turning response.


Immunological identification of high molecular weight forms common to bovine neurophysin and vasopressin.
Extracts of bovine neurohypophysis made in acid/ethanol solution containing protease inhibitors were fractionated by two successive filtrations on Sephadex G-75 columns equilibrated in the presence and then in the absence of 4 M urea. Analysis of the pattern of neurophysin-like immunoreactivity in the eluate, with two different antibodies, indicated the presence of high M(r) forms of neurophysin (apparent sizes, [unk]70,000 and 20,000-25,000, respectively) besides the M(r) 10,000 neurophysin. [8-Arginine]vasopressin-like immunoreactivity was also detected, coeluting with the neurophysin-like species, in the material recovered in the exclusion and M(r) 20,000-25,000 elution volumes of the same molecular sieve fractionation of neurohypophyseal extracts. Upon subsequent Sephadex G-150 filtration, the immunoreactive material recovered in the exclusion volume of the Sephadex G-75 filtration showed an apparent M(r) of approximately 140,000. Both neurophysin-like and vasopressin-like immunoreactivities coeluted in the same volume. The elution profile of this M(r) 140,000 material was unmodified when reanalyzed by the same molecular sieve filtration after exposure to 8 M urea. When these M(r) 140,000 immunoreactive forms of vasopressin and neurophysin were submitted to affinity chromatography on anti-neurophysin antibodies immobilized on Sepharose, both immunoreactivities were selectively coadsorbed to the immunoadsorbent. Similarly, the neurophysin and vasopressin immunoreactivities associated with M(r) approximately 25,000 were retained together on the same anti-neurophysin immunoadsorbent. The M(r) 140,000 and M(r) 25,000 species having both neurophysin and [8-arginine]vasopressin antigenic determinants generated the two neurosecretory components when exposed to proteolytic activities. This in vitro processing was inhibited in acid medium, at low temperature, and in the presence of a mixture of protease inhibitors. It is concluded that these two large forms of proteins containing both neurophysin and vasopressin may represent common biosynthetic precursors of these two neurohypophyseal components.


Characterization of O-mannosyltransferase family in Schizosaccharomyces pombe.
Protein O-glycosylation is an essential protein modification in eukaryotic cells. In Saccharomyces cerevisiae, O-mannosylation is initiated in the lumen of the endoplasmic reticulum by O-mannosyltransferase gene products (Pmt1p-7p). A search of the Schizosaccharomyces pombe genome database revealed a total of three O-glycoside mannosyltransferase homologs (ogm1+, ogm2+, and ogm4+), closely related to Saccharomyces cerevisiae PMT1, PMT2, and PMT4. Although individual ogm genes were not found to be essential, ogm1Delta and ogm4Delta mutants exhibited aberrant morphology and failed to agglutinate during mating. The phenotypes of the ogm4Delta mutant were not complemented by overexpression of ogm1+ or ogm2+, suggesting that each of the Ogm proteins does not have overlapping functions. Heterologous expression of a chitinase from S. cerevisiae in the ogm mutants revealed that O-glycosylation of chitinase had decreased in ogm1Delta cells. A GFP-tagged Fus1p from S. cerevisiae was specifically not glycosylated and accumulated in the Golgi in ogm4Delta cells. These results indicate that O-glycosylation initiated by Ogm proteins plays crucial physiological roles and can serve as a sorting determinant for protein transport of membrane glycoproteins in S. pombe.


Analysis of noise and bias in fermentation oxygen uptake rate data.
The calculation of many derived fermentation variables such as the respiratory quotient (RQ) and mass transfer coefficient (K(L)a) requires the differences between the molar percentages of oxygen and carbon dioxide in the fermentor inlet and exit gas, called the %OUR and %CER. Noise and bias in %CER data is of order that in the exit gas carbon dioxide analysis. However, the relative amount of noise in the %OUR is one to two orders of magnitude greater than the noise in the raw oxygen analyses because the %OUR is calculated as a small difference between two large quantities. The noise in the %OUR is white with a Gaussian amplitude probability distribution of absolute standard deviation 0.0145. A chi-square filter of the %OUR data is shown to considerably improve the quality of the calculated RQ and K(L)a for a fermentation of Streptomyces clavuligerus.


The ENZYME data bank.
The ENZYME data bank is a repository of information relative to the nomenclature of enzymes. It is primarily based on the recommendations of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB) and it contains the following data for each type of characterized enzyme for which an EC (Enzyme Commission) number has been provided: EC number Recommended name Alternative names (if any) Catalytic activity Cofactors (if any) Pointers to the SWISS-PROT protein sequence entrie(s) that correspond to the enzyme (if any) Pointers to human disease(s) associated with a deficiency of the enzyme (if any).


Single molecule as a local acoustic detector for mechanical oscillators.
A single molecule can serve as a nanometer-sized detector of acoustic strain. Such a nanomicrophone has the great advantage that it can be placed very close to acoustic signal sources and high sensitivities can be achieved. We demonstrate this scheme by monitoring the fluorescence intensity of a single dibenzoterrylene molecule in an anthracene crystal attached to an oscillating tuning fork. The characterization of the vibration amplitude and of the detection sensitivity is a first step towards detection and control of nanomechanical oscillators through optical detection and feedback.


[Letters of Sadaaki Kanezawa as resources for medical history].
Sadaaki Kanezawa was one of the people responsible for the Kamakura Hôjô feudal government. He is considered to have been a man of high cultural sense, and greatly contributed to bringing Kanezawa Shômyôji Temple and Kanezawa Bunko to prosperity. His letters to his family show his affection and love. He also left many letters to Ken-a, the second Elder of Kanezawa Shômyôji Temple. These letters relate the names of diseases of that time and different kinds of treatments and care, folk remedies, and the like. They are highly valuable materials for the study of medical history in Japan.


Genome sequence of Perigonia lusca single nucleopolyhedrovirus: insights into the evolution of a nucleotide metabolism enzyme in the family Baculoviridae.
The genome of a novel group II alphabaculovirus, Perigonia lusca single nucleopolyhedrovirus (PeluSNPV), was sequenced and shown to contain 132,831 bp with 145 putative ORFs (open reading frames) of at least 50 amino acids. An interesting feature of this novel genome was the presence of a putative nucleotide metabolism enzyme-encoding gene (pelu112). The pelu112 gene was predicted to encode a fusion of thymidylate kinase (tmk) and dUTP diphosphatase (dut). Phylogenetic analysis indicated that baculoviruses have independently acquired tmk and dut several times during their evolution. Two homologs of the tmk-dut fusion gene were separately introduced into the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) genome, which lacks tmk and dut. The recombinant baculoviruses produced viral DNA, virus progeny, and some viral proteins earlier during in vitro infection and the yields of viral occlusion bodies were increased 2.5-fold when compared to the parental virus. Interestingly, both enzymes appear to retain their active sites, based on separate modeling using previously solved crystal structures. We suggest that the retention of these tmk-dut fusion genes by certain baculoviruses could be related to accelerating virus replication and to protecting the virus genome from deleterious mutation.


Exploring predictors of HIV-1 virologic failure to long-acting cabotegravir and rilpivirine: a multivariable analysis.
Efficacy and safety of long-acting cabotegravir (CAB) and rilpivirine (RPV) dosed intramuscularly every 4 or 8 weeks has been demonstrated in three Phase 3 trials. Here, factors associated with virologic failure at Week 48 were evaluated post hoc. Data from 1039 adults naive to long-acting CAB+RPV were pooled in a multivariable analysis to examine the influence of baseline viral and participant factors, dosing regimen and drug concentrations on confirmed virologic failure (CVF) occurrence using a logistic regression model. In a separate model, baseline factors statistically associated with CVF were further evaluated to understand CVF risk when present alone or in combination. Overall, 1.25% (n = 13/1039) of participants experienced CVF. Proviral RPV resistance-associated mutations (RAMs), HIV-1 subtype A6/A1, higher BMI (associated with Week 8 CAB trough concentration) and lower Week 8 RPV trough concentrations were significantly associated (P < 0.05) with increased odds of CVF (all except RPV trough are knowable at baseline). Few participants (0.4%) with zero or one baseline factor had CVF. Only a combination of at least two baseline factors (observed in 3.4%; n = 35/1039) was associated with increased CVF risk (25.7%, n = 9/35). CVF is an infrequent multifactorial event, with a rate of approximately 1% in the long-acting CAB+RPV arms across Phase 3 studies (FLAIR, ATLAS and ATLAS-2M) through Week 48. Presence of at least two of proviral RPV RAMs, HIV-1 subtype A6/A1 and/or BMI at least 30 kg/m2 was associated with increased CVF risk. These findings support the use of long-acting CAB+RPV in routine clinical practice.


Treatment Modalities for Esophageal Adenocarcinoma in the United States: Trends and Survival Outcomes.
Background: Owing to the rising incidence of esophageal adenocarcinoma (EAC), we sought to assess the national trends in the use of different treatment modalities and compare survival outcomes among them. Methods: Using Surveillance, Epidemiology, and End Results (SEER) Program registry (2004-2014), we identified adult patients diagnosed with EAC undergoing definitive chemoradiotherapy (dCRT), esophagectomy, or neoadjuvant chemoradiotherapy plus esophagectomy (nCRT/S). Linear trends in the yearly incidence of each treatment were assessed using Poisson regression. An inverse probability of treatment weighted (IPTW) Cox regression was used to estimate the effect of each treatment on mortality. IPTW was used to account for potential confounding by year of diagnosis, patient demographics, and cancer characteristics. Results: A total of 10,755 patients were included in the study. From 2004 to 2014, the use of esophagectomy alone decreased from 15% to 5% (P < .0001), whereas nCRT/S increased from 14% to 20% (P < .0001); dCRT remained relatively stable (26% to 29%, P = .08). The 60-month survival rate was 13.0% for dCRT, 33.0% for esophagectomy only, and 36.3% for nCRT/S. After accounting for patient and cancer characteristics, both esophagectomy (hazard ratio [HR] 0.62, 95% confidence interval [CI] 0.55-0.70, P < .0001) and nCRT/S (HR 0.45, 95% CI 0.41-0.48, P < .0001) had significantly the highest survival rates. Conclusion: The use of esophagectomy alone has decreased, whereas nCRT/S has increased among EAC patients. Considering the better outcomes achieved with surgical resection, the use of dCRT should be discouraged in surgically fit patients.


Halothane anesthesia decreases the extracellular level of dopamine in rat striatum: a microdialysis study in vivo.
In our previous microdialysis study, sevoflurane or isoflurane anesthesia significantly decreased the extracellular level of dopamine in rat striatum in vivo. On the other hand, other investigators demonstrated that halothane anesthesia either increased or did not affect the extracellular dopamine level. To explore the differences among these volatile anesthetics, the effects of halothane and nitrous oxide on the striatal dopamine level were reinvestigated. Halothane alone, nitrous oxide with or without halothane, or drugs known to affect the dopaminergic pathway were administered to rats. Microdialysates were collected every 20 min and directly applied to an on-line high-performance liquid chromatograph without any pretreatment. The effects of halothane on respiratory and cardiovascular variables were monitored. General anesthesia with halothane alone decreased the dialysate (extracellular) concentration of dopamine but increased that of dopamine metabolites. Nitrous oxide alone slightly increased dopamine metabolites in dialysates but did not affect the halothane-induced decrease in extracellular dopamine. Apomorphine and haloperidol reproduced reported results, confirming the adequacy of our methodology. Nomifensine- or methamphetamine-induced increase in extracellular dopamine was augmented by halothane. These results suggest that halothane potently enhances striatal dopamine release and activates the reuptake or metabolic process, which is consistent with our previous results for sevoflurane or isoflurane. Volatile anesthetics interfere with dopamine regulation, at least in the rat striatum.


Open dose-finding study of a new potent and selective nonsteroidal aromatase inhibitor, CGS 20 267, in healthy male subjects.
The aim of this open, dose-finding study was to evaluate the effects of single dose CGS 20 267, a new oral nonsteroidal aromatase inhibitor, on the inhibition of estrogen production and also on the production of adrenal and testicular steroids in healthy male subjects. Nine dose levels ranging from 0.02-30 mg and placebo were tested, each dose being given to 3 subjects only. A total of 18 subjects were included; 12 of them received 2 single administration, the remaining 6 were exposed only once to one of the 2 highest dose levels. A reduction in serum estrogen levels when compared to baseline was already observed after 2 h, reaching maximum suppression between 10 and 48 h after administration. After 24 h, a suppression of estrone levels by 60-85% from baseline was achieved with all tested doses. A reduction in estradiol levels by about 30% from baseline was observed at the lowest dose (0.02 mg). This reduction was further enhanced dose dependently to a maximum of about 90% from baseline at 24 h after administration of the highest dose (30 mg). With the higher doses (10 and 30 mg), estrogen suppression was maintained up to 3 days. A dose-dependent increase of testosterone, LH, and FSH was observed and was most pronounced in the 10- and 30-mg dose groups, which can be considered as a consequence of the long-lasting aromatase inhibition achieved with these high doses. No effect on serum cortisol and aldosterone levels was observed up to the highest dose. No clinically relevant changes were observed in blood chemistry and hematology tests. The systemic and subjective tolerability of CGS 20 267 was good at all doses. This study has shown that CGS 20 267 is a well tolerated, potent, selective, and long-acting inhibitor of the aromatase enzyme after single administration.


Sleep Apnea Morbidity: A Consequence of Microbial-Immune Cross-Talk?
OSA has emerged as a highly prevalent public health problem that imposes important mid- and long-term consequences, namely cardiovascular, metabolic, cognitive, and cancer-related alterations. OSA is characterized by increased upper airway resistance, alveolar hypoventilation, and recurrent upper airway obstruction during sleep. Recurrent collapse of the upper airway develops with sleep onset and is associated with both intermittent hypoxemia and sleep fragmentation. The microbiome is a vast and complex polymicrobial ecosystem that coexists with the human organism, and it has been identified as playing significant roles in the development of host immunologic phenotypes. In humans and animal models, changes in gut microbial communities occur with lifestyle behaviors, such as smoking, long-distance travel, dietary preferences, physical exercise, and circadian rhythm disturbances. In parallel, diseases previously attributed in part to lifestyle such as obesity, coronary heart disease, depression, and asthma (also associated with OSA) are now claimed as microbiota related. We therefore posit that altered patterns of sleep and oxygenation, as seen in OSA, will promote specific alterations in gut microbiota that in turn will elicit the immunologic alterations that lead to OSA-induced end-organ morbidities. The present article assesses the potential mechanistic links between OSA-induced changes in gut microbiota and its morbid phenotypes.


Clinical utility of non-EpCAM based circulating tumor cell assays.
Methods enabling the isolation, detection, and characterization of circulating tumor cells (CTCs) in blood have clear potential to facilitate precision medicine approaches in patients with cancer, not only for prognostic purposes but also for prediction of the benefits of specific therapies in oncology. However, current CTC assays, which capture CTCs based on expression of epithelial cell adhesion molecule (EpCAM), fail to capture cells from de-differentiated tumors and carcinomas undergoing loss of the epithelial phenotype during the invasion/metastatic process. To address this limitation, many groups are developing non-EpCAM based CTC assays that incorporate nanotechnology to improve test sensitivity for rare but important cells that may otherwise go undetected, and therefore may improve upon clinical utility. In this review, we outline emerging non-EpCAM based CTC assays utilizing nanotechnology approaches for CTC capture or characterization, including dendrimers, magnetic nanoparticles, gold nanoparticles, negative selection chip or software-based on-slide methods, and nano-scale substrates. In addition, we address challenges that remain for the clinical translation of these platforms.


[Ainhum].
Ainhum is an ancient disease described in 1821 by Messum and its origin is still unknown. It is a tropical or semitropical disease seen in Blacks, Whites and Indians which affects the 5th and sometimes the 4th toe and causes spontaneous amputation. Females are more often affected than males, and there is a tendency to geographical clustering of cases and familial occurrence.


The sites of action of praziquantel in a smooth muscle of Lymnaea stagnalis.
Praziquantel (1.6 X 10(-6) M), a new anthelmintic drug, was shown to produce a sustained contracture in the penis retractor muscle of the freshwater snail Lymnaea stagnalis. Calcium-free saline greatly reduced the magnitude and duration of the praziquantel contractures if the preparation had been previously exposed to praziquantel. The praziquantel contracture in zero-calcium saline was abolished if the preparation was pretreated with successive applications of 2 X 10(-2) M caffeine. Lanthanum (1.0 mM) in zero-calcium saline prevented the contractures normally produced by 30 mM potassium saline, but did not prevent the praziquantel contracture. In normal saline, however, 1.0 mM lanthanum greatly reduced the magnitude of a praziquantel-induced contracture. We conclude that praziquantel increases sarcolemmal calcium permeability and releases calcium from intracellular stores in Lymnaea stagnalis smooth muscle.


Characterization of a Sequential UV Photolysis-Biodegradation Process for Treatment of Decabrominated Diphenyl Ethers in Sorbent/Water Systems.
Decabrominated diphenyl ether (BDE-209) is a primary component of the brominated flame retardants used in a variety of industrial and domestic applications. BDE-209 bioaccumulates in aquatic organisms and has been identified as an emerging contaminant that threatens human and ecosystem health. Sequential photolysis-microbial biodegradation processes were utilized here to treat BDE-209 in clay- or soil-water slurries. The removal efficiency of BDE-209 in the clay-water slurries was high; i.e., 96.5%, while that in the soil-water slurries was minimal. In the clay-water slurries the first order rate constants for the UV photolysis and biodegradation of BDE-209 were 0.017 1/day and 0.026 1/day, respectively. UV wavelength and intensity strongly influenced the BDE-209 photolysis and the subsequent biodegradation of photolytic products. Facultative chemotrophic bacteria, including Acidovorax spp., Pseudomonas spp., Novosphingobium spp. and Sphingomonas spp., were the dominant members of the bacterial community (about 71%) at the beginning of the biodegradation; many of these organisms have previously been shown to biodegrade BDE-209 and other polybrominated diphenyl ether (PBDE) congeners. The Achromobacter sp. that were isolated (NH-2; NH-4; NH-6) were especially effective during the BDE-209 degradation. These results indicated the effectiveness of the sequential UV photolysis and biodegradation for treating certain BDE-209-contaminated solids; e.g., clays; in bioreactors containing such solids as aqueous slurries. Achieving a similar treatment effectiveness for more heterogeneous solids containing natural organic matter, e.g., surface solids, appears to be significantly more difficult. Further investigations are needed in order to understand the great difference between the clay-water or soil-water slurries.


In vitro and in vivo microbiological evaluation of Cephapirin, a new antibiotic.
The microbiological properties of cephapirin, a new semisynthetis cephalosporin, have been studied. This antibiotic, if compared with ampicillin, shows a greater activity against gram positive bacteria, a lack of sensitivity to staphlococcal beta-lactamase, and a lower sensitivity to those produced by gram-negative bacteria. Useful therapeutic of cephapirin levels can be detected in human serum 6 h after administration of 500 and 1,000 mg parenterally.


Carbonic anhydrase IX in renal cell carcinoma: implications for prognosis, diagnosis, and therapy.
The clinical management of patients with renal cell carcinoma (RCC) remains difficult, and the development of new diagnostic, prognostic, and therapeutic tools is still required. To review the current knowledge on the RCC-associated antigen carbonic anhydrase IX (CAIX) and provide evidence for how this antigen may aid in the clinical management of RCC. Clinical papers describing diagnostic, prognostic, and/or therapeutic applications of CAIX in RCC were selected from the Pubmed database. The search was manually augmented by reviewing the reference lists of articles. Expression of CAIX is regulated by the Von Hippel Lindau (VHL) protein (pVHL). Because of the invariable VHL mutational loss in clear-cell RCC (ccRCC) patients, CAIX expression is ubiquitous in ccRCC. Determination of CAIX expression in nephrectomy specimens of RCC patients improves prognostic accuracy; high CAIX expression appears to correlate with a favourable prognosis and a greater likelihood of response to systemic treatment for metastatic disease. Therefore, CAIX expression might be used to stratify metastatic ccRCC (mRCC) patients for systemic treatment. When incorporated into the RCC nomogram, CAIX expression seems to improve diagnostic accuracy for primary RCC as well as mRCC patients, but further evidence is required. Clinical studies with the CAIX-specific monoclonal antibody (mAb) cG250 have provided unequivocal evidence that ccRCC lesions can be imaged with radiolabeled cG250. Results are awaited of a large, randomised trial that aims to establish the value of cG250 imaging for primary RCC. The outcome of another large, placebo-controlled study is awaited to establish the usefulness of CAIX-targeted therapy in the adjuvant setting. Therapeutic trials with high-dose radiolabeled cG250 and CAIX-loaded dendritic cells in mRCC patients are still in phase 1 or 2. CAIX improves diagnostic accuracy and is an attractive target for imaging of and therapy for ccRCC.


The use of individually wrapped presterilized small orthopaedic implants increase operating time: a prospective experimental study.
There have been concerns about the potential increases in operating time associated with the use of individually wrapped presterilized small orthopaedic implants compared with our traditional method of screw banks. We set out to quantify this theory. Prospective experimental study. Theatre. Orthopaedic Surgical Trainees and Theatre Scrub team. The time taken to complete the operation. The use of prepacked and sterilized implants added 2 min 56 s to the use of a bank with a full complement of normal screws that required tapping and 3 min 58 s if self-tapping screws were used (P < 0.001). Using individually wrapped presterilized small orthopaedic implants increases operating time.


Man o' war mutation in UDP-α-D-xylose synthase favors the abortive catalytic cycle and uncovers a latent potential for hexamer formation.
The man o' war (mow) phenotype in zebrafish is characterized by severe craniofacial defects due to a missense mutation in UDP-α-d-xylose synthase (UXS), an essential enzyme in proteoglycan biosynthesis. The mow mutation is located in the UXS dimer interface ∼16 Å away from the active site, suggesting an indirect effect on the enzyme mechanism. We have examined the structural and catalytic consequences of the mow mutation (R236H) in the soluble fragment of human UXS (hUXS), which shares 93% sequence identity with the zebrafish enzyme. In solution, hUXS dimers undergo a concentration-dependent association to form a tetramer. Sedimentation velocity studies show that the R236H substitution induces the formation of a new hexameric species. Using two new crystal structures of the hexamer, we show that R236H and R236A substitutions cause a local unfolding of the active site that allows for a rotation of the dimer interface necessary to form the hexamer. The disordered active sites in the R236H and R236A mutant constructs displace Y231, the essential acid/base catalyst in the UXS reaction mechanism. The loss of Y231 favors an abortive catalytic cycle in which the reaction intermediate, UDP-α-d-4-keto-xylose, is not reduced to the final product, UDP-α-d-xylose. Surprisingly, the mow-induced hexamer is almost identical to the hexamers formed by the deeply divergent UXS homologues from Staphylococcus aureus and Helicobacter pylori (21% and 16% sequence identity, respectively). The persistence of a latent hexamer-building interface in the human enzyme suggests that the ancestral UXS may have been a hexamer.


Myotomal myogenesis of axial muscle in the sturgeon Acipenser baeri (Chondrostei, Acipenseriformes).
Compared to teleost fishes, a unique character of the myogenesis of the plesiomorphic A. baeri is the fusion of myoblasts derived from the somite, leading to the formation of multinucleate muscle lamellae. Then, the lamellae are converted into cylindrical muscle fibres. The mechanism of transformation of lamellae into fibres is still debatable. Early embryonic muscle growth is mainly due to the hypertrophy of somite-cell derived stock. After hatching, hypertrophic growth occurs parallel to hyperplastic growth. Proliferatively active mesenchymal cells, which migrate from the intermyotomal space into the myotomes, participate in both processes.


Adhesion of Bacillus subtilis and Pseudoalteromonas lipolytica to steel in a seawater environment and their effects on corrosion.
In a marine environment, Bacillus subtilis and Pseudoalteromonas lipolytica are commonly found in the biofilms adherent to low-alloy engineering steel, and they have distinct effects on corrosion. In the present work, this phenomenon was investigated through the study of various materials characterization methods, electrochemical techniques, and contact angle measurements. It was found that the surface film formed on the steel in the presence of B. subtilis was compact, uniform, free of cracks, and hydrophobic. However, the film formed in the presence of P. lipolytica was loose, rough, heterogeneous, and hydrophilic. The main components of the films formed in the presence of B. subtilis and P. lipolytica were polysaccharides/TasA amyloid fibers and proteins/carboxylic acid, respectively. The composition, structure, and properties of the surface films formed on the steel were associated with different effects on corrosion. The presence of B. subtilis enhances the steel's resistance to corrosion, whereas corrosion was increased by the presence of P. lipolytica. In short, the compact and hydrophobic biofilm of B. subtilis appears to inhibit the corrosion of steel, while the loose, hydrophilic film of P. lipolytica tends to induce pitting corrosion.


Pathogenicity of anaerobic gram-positive cocci.
The pathogenicity of 20 strains of facultative or anaerobic gram-positive cocci (AGPC) was investigated by injecting them alone or mixed with other flora into mice, utilizing the subcutaneous abscess model. Abscesses induced by a mixture of two organisms were uniformly larger than those induced by single organisms. The relationships among seven AGPC strains, eight aerobes, and two Bacteroides spp. were determined by treating the infected animals with antibiotics and observing the effect of therapy directed against one or both organisms present in the abscess. A total of 70 different combinations were tested. As judged by their responses to antimicrobial therapy, facultative cocci or AGPC were relatively more important than the other species in 6 combinations, equally important in 35 combinations, and less important in 29 combinations. The AGPC most often found to be equal to or more important than the other bacteria were Peptococcus magnus, Streptococcus constellatus, and Peptostreptococcus anaerobius. Proteus mirabilis, Escherichia coli, and Staphylococcus aureus were frequently found to be of more importance than the AGPC.


GCK mutations in Chinese MODY2 patients: a family pedigree report and review of Chinese literature.
Maturity-onset diabetes of the young type 2 (MODY2) is caused by mutations in the glucokinase (GCK) gene and is rare in the Chinese population. We report three Chinese families with MODY2 and the sequencing of the GCK gene. Three unrelated Chinese families with MODY2 and their pedigrees were investigated. In Family 1, the proband was a 7-year-old girl with impaired fasting glucose (IFG) and impaired glucose tolerance (IGT). Her mother and maternal grandfather had IFG. In Family 2, the proband was a boy who had diabetes mellitus at 11 years. His sister had IFG. His father and grandmother had diabetes mellitus at 22 and 25 years, respectively. In Family 3, the proband was a boy who had IFG and IGT at 12 years. His sister had diabetes mellitus at 8 years. His father and grandfather had IFG and/or IGT. The GCK gene was directly sequenced. Diabetes mellitus or IFG/IGT was found among three consecutive generations in three families. One novel nonsense heterozygous mutation in exon 5 (c.556 C>T, p.Arg 186 stop) was detected in Family 1. Another novel frameshift mutation in exon 4 (c.367-374dupTTCGACTA, p.Ile 126 fs) was found in Family 2. A previously reported, a missense heterozygous mutation in exon 5 (c.571 C>T, p.Arg 191Trp) was detected in Family 3. The thorough investigation of three Chinese families with MODY2 revealed two novel mutations and one known mutation. GCK gene sequencing helps in MODY2, especially when there is uncertain IFG or IGT.


Pathogenesis and epidemiology of vulvovaginal candidiasis.
In contrast to women with infrequent episodes of candidal vaginitis, women with chronic and recurrent candidal vaginitis do not have recognizable precipitating or causal factors. Analysis of vaginal yeast isolated from women with recurrent candidal vaginitis, although revealing a higher percentage of non-albicans Candida sp., indicates that resistance to imidazoles is not a causal factor, and other fungal virulence factors that could explain the repeated attacks have not been identified. Strain typing of sequential clinical isolates indicates a pattern of vaginal relapse rather than frequent vaginal reinfection, and attempts to reduce attacks by treating sexual partners and suppressing a gastrointestinal tract focus have failed. Recent immunological studies suggest the possibility of an acquired Candida antigen-specific immunological deficiency resulting in uncontrolled vaginal Candida proliferation and hence repeated clinical attacks. In support of the immunological hypothesis is the recent report of normal T lymphocyte suppression of Candida hyphal production.


Does concomitant stress incontinence alter the efficacy of tolterodine in patients with overactive bladder?
Muscarinic antagonists such as tolterodine are the treatment of choice for overactive bladder (OAB). We determined the impact of concomitant stress incontinence (SI) on the therapeutic effects of tolterodine in patients with OAB with and without concomitant SI. Data from an open label, observational study involving 2,250 patients with OAB symptoms were analyzed for baseline frequency, urgency and incontinence, and alterations in these symptoms while on 12-week treatment with 2 mg tolterodine twice daily. Data are shown as the mean +/- SD. The statistical significance of differences in treatment effects was determined by multiple regression analysis, adjusting for gender, age and baseline symptom intensity. Concomitant I to III degree SI according to the Stamey grading was present in 31%, 15% and 2% of patients, respectively, and it was associated with increasing basal incontinence, although only III degree SI was associated with greater baseline frequency or urgency. In the overall group tolterodine decreased frequency, urgency and urge incontinence from 12.4 +/- 4.3 to 7.7 +/- 2.7, 8.4 +/- 5.1 to 2.0 +/- 3.0 and 3.4 +/- 4.2 to 0.8 +/- 2.0 episodes daily, respectively. On multiple linear regression analysis I and II degree SI had a minor, if any, effect on this improvement, while III degree SI was statistically associated with a smaller decrease in frequency (by 1.4 +/- 0.4 micturitions daily, p = 0.0002) and incontinence (by 2.1 +/- 0.3 episodes daily, p < 0.0001) but with similar alterations in the number of urge episodes. Concomitant I or II degree SI has little effect on the efficacy of tolterodine in OAB cases. Only patients with concomitant III degree SI have significantly less improvement.


Detecting Lung Diseases from Exhaled Aerosols: Non-Invasive Lung Diagnosis Using Fractal Analysis and SVM Classification.
Each lung structure exhales a unique pattern of aerosols, which can be used to detect and monitor lung diseases non-invasively. The challenges are accurately interpreting the exhaled aerosol fingerprints and quantitatively correlating them to the lung diseases. In this study, we presented a paradigm of an exhaled aerosol test that addresses the above two challenges and is promising to detect the site and severity of lung diseases. This paradigm consists of two steps: image feature extraction using sub-regional fractal analysis and data classification using a support vector machine (SVM). Numerical experiments were conducted to evaluate the feasibility of the breath test in four asthmatic lung models. A high-fidelity image-CFD approach was employed to compute the exhaled aerosol patterns under different disease conditions. By employing the 10-fold cross-validation method, we achieved 100% classification accuracy among four asthmatic models using an ideal 108-sample dataset and 99.1% accuracy using a more realistic 324-sample dataset. The fractal-SVM classifier has been shown to be robust, highly sensitive to structural variations, and inherently suitable for investigating aerosol-disease correlations. For the first time, this study quantitatively linked the exhaled aerosol patterns with their underlying diseases and set the stage for the development of a computer-aided diagnostic system for non-invasive detection of obstructive respiratory diseases.


Determination of birefringence and slow axis distribution using an interferometric measurement system with liquid crystal phase shifter.
It is known that liquid crystal (LC) cells are useful as compact and easy-to-handle phase shifters that are readily coupled into the optics of standard microscope systems. Here, a uniformly aligned molecular LC phase shifter is introduced into a polarization microscope to attain a birefringence imaging system, using the phase-shift interferometric technique. Since the birefringence can be determined accurately only when the optical axis of the sample is parallel or perpendicular to the slow axis (variable axis) of the LC phase shifter, an improved data analysis method is proposed for determining the birefringence independently of the direction; a simple method of determining the slow axis distribution is also demonstrated. Measurements of the birefringence and slow axis distribution properties of a potato starch particle are demonstrated to confirm the novel determination method.


Significant association between neutrophil aggregation in aspirated thrombus and myocardial damage in patients with ST-segment elevation acute myocardial infarction.
This study was designed to clarify the relationship between myocardial damage and platelet-neutrophil aggregation in patients with acute myocardial infarction (AMI). The study group comprised 107 patients with ST-segment elevation AMI, in whom aspiration catheter was used during emergency percutaneous coronary intervention. Patients were divided into 2 groups according to the cellular density of neutrophils in the aspirated sample: group L (n=53), <or=100 neutrophils/0.025 mm(2) thrombus; group H (n=54), >100 neutrophils/0.025 mm(2) thrombus. Myocardial blush grade (MBG) <or=1 and ST-segment resolution (STR) <50% were more frequently found in group H than in group L. Peak creatine kinase level tended to be higher and left ventricular ejection fraction (LVEF) at 6 months after onset was lower in group H than in group L. Multivariate analysis showed that high neutrophil density in aspirated thrombus was an independent predictor of MBG <or=1, STR <50%, and low LVEF at 6 months after onset. Platelet-neutrophil aggregates retrieved from ruptured plaque may be associated with impaired coronary microcirculation and resultant myocardial necrosis/dysfunction. These findings underscore the clinical importance of the interaction between thrombosis and inflammation in the pathogenesis of AMI.


[Assessments of sleepiness in adolescents: A key tool for better intervene in mental health].
Objectives Daytime sleepiness in adolescents has negative impacts on physical, cognitive, and emotional health, with direct or indirect consequences on their mental health. This review aims to describe specialized tools assessing daytime sleepiness in adolescents so that mental health professionals can screen for a variety of sleep disorders, from the rarest ones, such as narcolepsy, to the most common ones, such as sleep-wake cycle delay in adolescents. Method Articles were selected in Medline (https://pubmed.ncbi.nlm.nih.gov/) and targeted adolescents aged between 13 and 18 or the keyword "adolescent*". The keywords used were: "sleepiness test" AND "questionnaire*". Only articles in French or English and published until January 9, 2023 were included. A total of 277 scientific articles were screened. Final sample included a total of 35 articles describing sleepiness measurement tools in adolescents. Results Among the 35 articles, a total of seven daytime sleepiness measurement tools in adolescents were identified. Four of them were subjective: 1) the Epworth Sleepiness Scale for Children and Adolescents (ESS-CHAD), 2) the Pediatric Daytime Sleepiness Scale (PDSS), 3) the Cleveland Adolescent Sleepiness Questionnaire (CASQ) and 4) the French Sleepiness Scale for Adolescents (FSSA). These self-reported questionnaires are less expensive and they can be used easily by mental health professionals as opposed to objective tools. Three objective tools have been identified: 1) the multiple sleep latency test (MSLT), 2) the maintenance of wakefulness test or called the "Maintenance Wakefulness Test" (MWT) and 3) the pupillographic sleepiness test (PST). Conclusion Given that adolescents end-of the-day sleep pressure, often resulting in a greater opportunity to light exposure, they are more at risk for daytime sleepiness and consequently to mental health challenges. Mental health professionals should therefore systematically screen for daytime sleepiness in adolescents using subjective tools. There are reliable and validated tools that are translated into French, such as the FSSA and the ESS-CHAD to measure daytime sleepiness in adolescents and lifestyles problems associated with sleep loss When daytime sleepiness suggests the presence of medical-based sleep disorders, such as narcolepsy, restless sleep disorders or sleep apnea, it is important to pursue an investigation with objective tools (nocturnal polysomnography, MLST and MWT) in collaboration with the adolescent's physician.


Prognostic factors and nomogram for the overall survival of bladder cancer bone metastasis: A SEER-based study.
Bone metastasis has a poor prognosis in patients with bladder cancer (BC). This study aimed to construct a prognostic nomogram for predicting the overall survival of patients with bone-metastatic BC (BMBC). The Surveillance, Epidemiology, and End Results database was used to recruit patients with BMBC between 2010 and 2018. Univariate and multivariate analyses were performed to screen for prognostic factors and construct a nomogram. Harrell concordance index, receiver operating characteristic curve, and calibration curve were used to verify the prognostic nomograms. All statistical analyses and chart formation were performed using SPSS 23.0 and R software 4.1.2. A total of 1361 patients diagnosed with BMBC were identified in the Surveillance, Epidemiology, and End Results database. Six independent prognostic factors, including marital status, histological type, T stage, other metastases, surgery, and chemotherapy, were identified and included in the nomogram construction. Among them, chemotherapy contributed the most to the prognosis in the nomogram. The concordance index of the nomogram was 0.745 and 0.753 in the training and validation groups, respectively, and all values of the area under the curve were >0.77. The calibration curves showed perfect consistency between the observed and predicted survival rates. The prognostic nomogram developed in this study is expected to become an accurate and individualized tool for predicting overall survival in patients with BMBC and providing guidance for appropriate treatment or care.


Spatial localization ability of planarians identified through a light maze paradigm.
Spatial localization ability is crucial for free-living animals to fit the environment. As shown by previous studies, planarians can be conditioned to discriminate directions. However, due to their simplicity and primitiveness, they had never been considered to have true spatial localization ability to retrieve locations of objects and places in the environment. Here, we introduce a light maze training paradigm to demonstrate that a planarian worm can navigate to a former recognized place from the start point, even if the worm is transferred into a newly produced maze. This finding identifies the spatial localization ability of planarians for the first time, which provides clues for the evolution of spatial learning. Since the planarians have a primitive brain with simple structures, this paradigm can also provide a simplified model for a detailed investigation of spatial learning.


Oral Health Knowledge, Attitudes and Behaviour of Parents and Caregivers of Preschool Children: Implications for Oral Health Promotion.
The aim of this study was to describe oral health knowledge, attitudes and behaviours of parents and caregivers of preschool children in order to inform an oral health promotion strategy. A sample of parents and caregivers of children attending nine randomly selected preschools in central Trinidad were invited to complete a self-administered questionnaire on early childhood oral health. A total of 309 parents and caregivers participated: 88% were female, 74.4% were of Indian ethnicity, with 50.4% in manual employment, and 50.2% educated to secondary level. 59.1% felt a child's first dental visit should be when all primary teeth are present. 64% had not taken their child for a dental visit. 81.6% rated their child's oral health as good or better and 28% would want an asymptomatic, decayed primary tooth extracted rather than filled. Over 80% used fluoride toothpaste. 52.8% always supervised their child's toothbrushing, and 44% claimed to be using the recommended pea-size amount. 26.2% reported having used a sweetened feeding bottle or infant feeder at night. Parents and caregivers of preschool children in this sample had reasonable oral health knowledge. However, despite generally positive attitudes towards preventive oral healthcare, confusion regarding dental attendance, supervised toothbrushing, fluoride use and sugar intake suggests that these items require particular emphasis in oral health promotion programmes aimed at improving early childhood oral health.


scribble (scrib) knockdown induces tumorigenesis by modulating Drp1-Parkin mediated mitochondrial dynamics in the wing imaginal tissues of Drosophila.
scrib loss of function is associated with various human-cancers. Most of the human-cancers have been characterized by mitochondrial dysfunction with elevated oxidative stress. However, the role of scrib to mitochondrial dysfunction in cancer has not been investigated earlier. Here, we have shown that scrib knockdown leads to mitochondrial depolarization, fragmentation and perinuclear-clustering along with disruption of the redox homeostasis. Moreover, the scrib abrogated tumor showed the elevation of Drp-1 and reduced expression of Marf, which suggests enhanced mitochondrial-fission. Further, the reduced expression of Parkin and HtrA2 interpret defective mitophagy leading to clustering of fragmented mitochondria and apoptotic inhibition in scrib knockdown tumors. Also, Parkin immunostaining depicted its reduced expression and mislocalization in the tumor cells in comparison to wild type. Moreover, the genetic study revealed the epistatic interactions of parkin and scrib. Thus, for the first time our results suggested that scrib loss induced mitochondrial-dysfunction modulates cancer progression by altering the mitochondrial dynamics regulators.


Synthesis and biological activity of 2'-deoxy-6-methyl-5-azacytidine and its alpha-D-anomer.
The stannic chloride catalyzed glycosylation of bis-tri-methylsilyl-6-methyl-5-azacytosine 2 with the halogenose 3 leading to the protected anomeric nucleosides 4a and 4b was investigated. Methanolysis of 4a and 4b afforded the corresponding free nucleosides 1a and 1b. Compounds 4a and 4b were also prepared by the isocyanate method via acetylamidinourea derivatives 6. Antileukemic activity in vitro and inhibition of growth of E. coli by the title compounds are reported.


Are Routine Chest Radiographs After Chest Tube Removal in Thoracic Surgery Patients Necessary?
The routine use of chest x-ray (CXR) to evaluate the pleural space after chest tube removal is a common practice driven primarily by surgeon preference and institutional protocol. The results of these postpull CXRs frequently lead to additional interventions that serve only to increase health care costs and resource utilization. We investigated the utility of these postpull CXRs in thoracic surgery patients and assessed their effectiveness in predicting the need for tube replacement. Single-institution retrospective study comprising thoracic surgery patients requiring postoperative chest tube drainage over a 3-y period. Demographics and surgical characteristics, including surgical approach, procedure, and procedure type, were recorded. Outcomes included postpull CXR findings, interventions resulting from radiographic abnormalities, and the additional health resource utilization incurred by obtaining these studies on asymptomatic patients. The study included 433 patients. Postpull CXRs were performed in 87.1% of patients, with 33.2% demonstrating an abnormality compared with the prior study. Among these, 65.7% resulted only in repeat imaging and 25.7% resulted in discharge delay. Overall, a total of 13 patients (3%) required chest tube replacement, three during the index hospitalization and the other 10 requiring readmission. Among those requiring chest tube replacement, 75% had normal postpull imaging, and all were symptomatic. Recurrent pneumothorax after chest tube removal requiring immediate tube reinsertion is relatively rare and does not occur in the absence of symptoms. Our study suggests that routine postpull CXRs have limited clinical utility and can be safely omitted in asymptomatic patients with appropriate clinical observation.


[Modern technologies and prospects of rehabilitation of patients after ischemic stroke].
Despite the great achievements in the field of neurorehabilitation, a significant proportion of patients after an ischemic stroke have persistent motor disturbances even after timely and adequately carried out restorative measures. The article discusses the issues of neuroplasticity, modern diagnostic technologies for studying this phenomenon; prognostic factors for recovery deficit following stroke and determining the effectiveness of ongoing treatment. The principles of neuroprotective therapy in ischemic stroke are considered, which is a pathogenetically justified direction at all stages of restorative treatment after cerebral circulation disorders. One of the most studied original cytoprotectors, demonstrating safety, efficacy and good tolerability, is cytoflavin. The results of numerous clinical trials have revealed a significant positive clinical and morphological dynamics when taking cytoflavin in patients after ischemic stroke.


From tumors to species: a SCANDAL hypothesis.
: Some tumor cells can evolve into transmissible parasites. Notable examples include the Tasmanian devil facial tumor disease, the canine transmissible venereal tumor and transmissible cancers of mollusks. We present a hypothesis that such transmissible tumors existed in the past and that some modern animal taxa are descendants of these tumors. We expect potential candidates for SCANDALs (speciated by cancer development animals) to be simplified relatives of more complex metazoans and have genomic alterations typical for cancer progression (such as deletions of universal apoptosis genes). We considered several taxa of simplified animals for our hypothesis: dicyemida, orthonectida, myxosporea and trichoplax. Based on genomic analysis we conclude that Myxosporea appear to be the most suitable candidates for a tumor ancestry. They are simplified parasitic cnidarians that universally lack major genes implicated in cancer progression including all genes with Caspase and BCL2 domains as well as any p53 and apoptotic protease activating factor - 1 (Apaf-1) homologs, suggesting the disruption of main apoptotic pathways in their early evolutionary history. Further comparative genomics and single-cell transcriptomic studies may be helpful to test our hypothesis of speciation via a cancerous stage. REVIEWERS: This article was reviewed by Eugene Koonin, Mikhail Gelfand and Gregory M Woods.


Coexpression of Argonaute-2 enhances RNA interference toward perfect match binding sites.
RNAi is widely applied to inhibit expression of specific genes, but it is limited by variable efficiency and specificity of empirically designed siRNA or shRNA constructs. This complicates studies targeting individual genes and significantly impairs large-scale screens using genome-wide knockdown libraries. Here, we show that ectopic expression of the RISC slicer Argonaute-2 (Ago2, eIF2C2) dramatically enhances RNAi specifically for mRNA targets with perfectly matched binding sites. This effect depends on its endonuclease activity and is uncoupled from its regulation of microRNA expression. To model the application of Ago2 coexpression with shRNA knockdown, we targeted the EGF receptor (EGFR) in lung cancer cells exhibiting oncogene addiction to EGFR. Whereas multiple empirically designed shRNA constructs exhibited highly divergent efficiencies in mediating EGFR knockdown and cell killing, coexpression of Ago2 resulted in uniform and highly specific target gene suppression and apoptosis in EGFR-dependent cells. Codelivery of Ago2 with shRNA constructs or siRNA duplexes thus provides a strategy to enhance the efficacy and the specificity of RNAi in experimental and potentially therapeutic settings.


Angelica tenuissima Nakai Ameliorates Cognitive Impairment and Promotes Neurogenesis in Mouse Model of Alzheimer's Disease.
To research Angelica tenuissima Nakai (ATN) for use in novel Alzheimer's disease (AD) therapeutics. The effect of a 30% ethanol extract of ATN (KH032) on AD-like cognitive impairment and neuropathological and neuroinflammatory changes induced by bilateral intracerebroventricular injections of β-amyloid (Aβ) peptide (Aβ1-42) was investigated. Male C57Bl/6 mice were randomly divided into 4 groups, 10 in each group. KH032-treated groups were administrated with a low or high dose of KH032 (50 and 200 mg/kg, respectively), intragastrically for 16 days; distilled water was applied in the sham and negative groups. Open fifield test, Y maze and Morris water maze test were used for behavior test and cognitive ability. In addition, the neuroprotective effects of KH032 in Aβ1-42-infused mice on the histopathological markers [neuronspecific nuclear protein (NeuN), Aβ1-42] of neurodegeneration were examined. The levels of glial fibrillary acidic protein (GFAP), NeuN, phosphorylation extracellular signal-regulated kinase (ERK)/ERK, brain-derived neurotrophic factor (BDNF), phosphorylation cAMP response element-binding (CREB)/CREB protein expression were measured by Western blot. KH032 treatment ameliorated cognitive impairments, reduced the overexpression of Aβ1-42, and inhibited neuronal loss and neuroinflammatory response in the Aβ1-42-infused mice. Moreover, KH032 treatment enhanced BDNF expression levels in the hippocampus. Finally, KH032 treatment increased phosphorylation of ERK1/2 and CREB, vital for ERK-CREB signaling. KH032 attenuated cognitive defificits in the Aβ1-42-infused mice by increasing BDNF expression and ERK1/2 and CREB phosphorylation and inhibiting neuronal loss and neuroinflflammatory response, suggesting that KH032 has therapeutic potential in neurodegenerative disorders such as AD.


Keyhole limpet hemocyanin (KLH)--a potent antigen in reaginic sensitization of mice and rats; a comparison with egg albumin (EA)-induced reaction.
Two distinct antigenic functions of KLH and EA were tested in four inbred strains of mice (BALB/c, 129, C3H/A, C57BL/6J:a) stimulation of reaginic antibody production after subcutaneous introduction of antigen in alum-precipitated form, and b) ability to elicit an anaphylactic reaction in vitro on pre-sensitized peritoneal mast cells. KLH was found to be more effective than EA in both functions tested, inducing higher titres of IgE and IgG1 antibody levels in the sera as well as higher percentage of histamine release from actively and passively sensitized peritoneal mast cells. Two different forms of KLH (associated and dissociated) were tested for potency in stimulating an anaphylactic sensitization of mice and rats and capacity to elicit an anaphylactic reaction on their peritoneal mast cells in vitro. In all rats as well as in BALB/c, 129 and C3H/A mice sensitization with dissociated KLH resulted in higher percentage of antigen-induced histamine release from peritoneal mast cells (an exception: mast cells of C57CL/6J mice released a little more histamine on the challenge with associated KLH). The PCA tests show, that the dissociated KLH does not act through greater stimulation of antibody formation.


Dcr2 targets Ire1 and downregulates the unfolded protein response in Saccharomyces cerevisiae.
Accumulation of unfolded proteins in the endoplasmic reticulum triggers the unfolded protein response (UPR). How the UPR is downregulated is not well understood. Inositol requirement 1 (Ire1) is an endoplasmic reticulum transmembrane UPR sensor in Saccharomyces cerevisiae. When the UPR is triggered, Ire1 is autophosphorylated, on Ser 840 and Ser 841, inducing the cytosolic endonuclease activity of Ire1, thereby initiating the splicing and translational de-repression of HAC1 mRNA. Homologous to Atf/Creb1 (Hac1) activates UPR transcription. Here, we report that the dose-dependent cell-cycle regulator 2 (Dcr2) phosphatase functionally and physically interacts with Ire1. We identified genetic interactions between DCR2 and genes, including IRE1, which are involved in secretory processes. Overexpression of DCR2, but not of a catalytically inactive DCR2 allele, significantly delays HAC1 splicing and sensitizes cells to the UPR. Furthermore, Dcr2 physically interacts in vivo with Ire1-S840E,S841E, which mimics phosphorylated Ire1, and Dcr2 de-phosphorylates Ire1 in vitro. Our results are consistent with de-phosphorylation of Ire1 being a mechanism for antagonizing UPR signalling.


[The induction of lymphokine-activated killer cells: increased cytotoxicity via long-term cultivation].
A long-time culture regime is tested for the production of lymphokine-activated killer cells from human lymphocytes, which combines the advantage of cell expansion with an increased cytotoxicity. The concentrations of IL-2 used are relatively low.


Neoplasms of the mononuclear phagocyte system: criteria for diagnosis.
The Mononuclear Phagocyte System, including promonocytes, monocytes, and tissue histiocytes (macrophages), gives rise to moncytic leukaemias and histiocytoses. A classification of these neoplasms by their presumed cell of origin is proposed, and criteria for their diagnosis are given. Histopathological, cytochemical, immunological and ultrastructural features useful in distinguishing these disorders form other haematopoietic or lymphoid neoplasms are presented in detail. The resulting classification reflects our knowledge at this time; as techniques become available to define further functional subpopulations of mononuclear phagocytes, it is likely that this classification will become as diverse as that of lymphoid neoplasms.


[Effects of salvianolic acid B on lipid peroxidation and metalloproteinase-2 activity in fibrotic liver in rat].
To investigate the mechanism of salvianolic acid B (Sal B) action against liver fibrosis through preventing lipid peroxidation and regulating MMP-2 activity in liver. The liver fibrotic model was induced through intraperitoneally injection of DMN at a dose of 10 microg x kg(-1) for every other day and lasting for 4 weeks. Sal B was administered (10 mg x kg(-1)), and perindopril (5 mg x kg(-1)) was used as positive control. Hepatic inflammation and collagen were observed with HE and sirius red staining. The liver function including serum ALT, AST activity, Alb and total bilirubin (T. Bil) level were determined. The hepatic lipid peroxidation including SOD and GST activities and GSH content were measured. Hepatic hydroxyproline (Hyp) content was detected with Jamall's method. The activity of metalloproteinase was assayed by gelatin zymography. The expressions of alpha-SMA, Col I in liver tissue were analyzed by Western blot. The model rats had higher serum T. Bil content, ALT and AST activities but lower Alb content than the normal rats, also had remarkable inflammatory necrosis and collagen deposition in liver, with much higher Hyp content, protein expression of alpha-SMA and collagen I and MMP-2 activity in liver, but had a decreased GSH content, SOD and GST activities. Both Sal B and perindopril attenuated hepatic injury and collagen deposition in model rats, decreased serum ALT activity and hepatic Hyp content, down-regulated alpha-SMA and collagen I protein expressions and metalloproteinase-2 activity than those in the model group, but increased SOD activity and GSH content, and Sal B decreased serum T. Bil content and increased GST activity. Sal B had a much better comprehensive actions than perindopril. Sal B has a good preventive action against liver fibrosis, the action mechanism is related to the prevention from lipid peroxidation and down-regulation of metalloproteinase-2 activity in fibrotic liver.


Expression of Molecular Differentiation Markers Does Not Correlate with Histological Differentiation Grade in Intrahepatic Cholangiocarcinoma.
The differentiation status of tumor cells, defined by histomorphological criteria, is a prognostic factor for survival of patients affected with intrahepatic cholangiocarcinoma (ICC). To strengthen the value of morphological differentiation criteria, we wished to correlate histopathological differentiation grade with expression of molecular biliary differentiation markers and of microRNAs previously shown to be dysregulated in ICC. We analysed a series of tumors that were histologically classified as well, moderately or poorly differentiated, and investigated the expression of cytokeratin 7, 19 and 903 (CK7, CK19, CK903), SRY-related HMG box transcription factors 4 and 9 (SOX4, SOX9), osteopontin (OPN), Hepatocyte Nuclear Factor-1 beta (HNF1β), Yes-associated protein (YAP), Epithelial cell adhesion molecule (EPCAM), Mucin 1 (MUC1) and N-cadherin (NCAD) by qRT-PCR and immunostaining, and of miR-31, miR-135b, miR-132, miR-200c, miR-221 and miR-222. Unexpectedly, except for subcellular location of SOX9 and OPN, no correlation was found between the expression levels of these molecular markers and histopathological differentiation grade. Therefore, our data point toward necessary caution when investigating the evolution and prognosis of ICC on the basis of cell differentiation criteria.


Anomalous Stretchable Conductivity Using an Engineered Tricot Weave.
Robust electric conduction under stretching motions is a key element in upcoming wearable electronic devices but is fundamentally very difficult to achieve because percolation pathways in conductive media are subject to collapse upon stretching. Here, we report that this fundamental challenge can be overcome by using a parameter uniquely available in textiles, namely a weaving structure. A textile structure alternately interwoven with inelastic and elastic yarns, achieved via a tricot weave, possesses excellent elasticity (strain up to 200%) in diagonal directions. When this textile is coated with conductive nanomaterials, proper textile engineering allows the textile to obtain an unprecedented 7-fold conductivity increase, with conductivity reaching 33,000 S cm(-1), even at 130% strain, due to enhanced interyarn contacts. The observed stretching conductivity can be described well using a modified 3D percolation theory that reflects the weaving effect and is also utilized for stretchable electronic interconnects and supercapacitors with high performance.


Effect of amendments on phytoavailability and fractionation of copper and zinc in a contaminated soil.
The ability of amendments to modify the soil properties and influence plants to immobilise Cu and Zn was studied in a naturally contaminated, additionally spiked podzolic soil. Lolium perenne L (perennial rye grass), Festuca rubra L (creeping red fescue) and Poa pratensis L (Kentucky blue grass) were tested in a pot study in the presence of soil amendments (lime, phosphate, and compost, individually and in combination) to assess the effect of soil-plant-amendment interaction on phytostabilisation. The ability of treatments to stabilize metals was assessed on the basis of metal fractionation in soil, partitioning of metals in plants, and metal uptake by the plants. Significant partitioning of Cu into immobile forms occurred as a result of the growth of Festuca rubra, and of Zn by the growth of Poa pratensis. Application of lime significantly reduced the exchangeable fraction of Zn, whereas phosphate application had an accelerating effect on exchangeable Cu. With combined application of amendments, the plant metal concentration decreased by more than 40% for Cu and 70% for Zn, compared to soils receiving no amendments. Combined application of amendments, in conjunction with growth of Festuca and Poa, can be recommended for phytostabilising of Cu and Zn in moderately contaminated acid soils of southwest British Columbia.


Alpha(1D)-adrenoceptors mediate nerve and agonist-evoked contractions in mouse vas deferens: evidence obtained from knockout technology.
1 It has been demonstrated that nerve-evoked contractions of the rat vas deferens involve alpha(1D)-adrenoceptors. Definitive evidence for a similar alpha(1D)-adrenoceptor-mediated response in mouse vas deferens has been more difficult to obtain. In this study, we have used alpha(1D)-adrenoceptor knockout (alpha(1D)-KO) mice to aid in the pharmacological characterization. 2 Mouse whole vas deferens was stimulated with a single pulse every 5 min. Once a stable response had been obtained, vehicle or antagonist was administered cumulatively at 5-min intervals and a response to stimulation obtained 5 min later. Cumulative concentration-response curves were also obtained for noradrenaline. 3 In vas deferens from alpha(1D)-KO mice, the contractile response to low concentrations of noradrenaline and the contractile response to a single stimulus were significantly reduced as compared to wild type (WT). 4 The alpha(1D)-adrenoceptor selective antagonist, BMY 7378, produced a concentration-dependent inhibition of single pulse-evoked contractions of vas deferens from WT and alpha(1D)-KO mice. BMY 7378 was significantly less potent in inhibiting stimulation-evoked contractions in vas deferens from alpha(1D)-KO mice. 5 It is concluded that alpha(1D)-adrenoceptors mediate a component of nerve- and agonist-evoked contractions of the vas deferens of WT mice.


Control of Collagen Triple Helix Stability by Phosphorylation.
The phosphorylation of the collagen triple helix plays an important role in collagen synthesis, assembly, signaling, and immune response, although no reports detailing the effect this modification has on the structure and stability of the triple helix exist. Here we investigate the changes in stability and structure resulting from the phosphorylation of collagen. Additionally, the formation of pairwise interactions between phosphorylated residues and lysine is examined. In all tested cases, phosphorylation increases helix stability. When charged-pair interactions are possible, stabilization via phosphorylation can play a very large role, resulting inasmuch as a 13.0 °C increase in triple helix stability. Two-dimensional NMR and molecular modeling are used to study the local structure of the triple helix. Our results suggest a mechanism of action for phosphorylation in the regulation of collagen and also expand upon our understanding of pairwise amino acid stabilization of the collagen triple helix.


Olfactory ensheathing glia injections in Beijing: misleading patients with ALS.
Different forms of cell transplantation therapy are being tested in models of ALS. While the approach offers hope to patients with ALS, much still needs to be learned in the laboratory before it is ready for human trials. Nevertheless, clinics across the world offer various types of open label cell transplantation therapy for high fees. We report a woman who received an injection into each frontal lobe in Beijing, China. Her ALS progressed at a more rapid rate after the procedure and she suffered disabling side-effects. Clinics that give experimental and potentially harmful treatments outside the construct of well-designed clinical trials put patients at risk and do a disservice to the ALS community.


Gloeolactone, a New Epoxy Lactone from a Blue-Green Alga
A novel lactone, gloeolactone (1), has been isolated from the blue-green alga, Gloeotrichia sp. The structure of this compound has been elucidated from a detailed analysis of the NMR spectra. This compound was shown to be toxic to brine shrimp.


Dosing schedule-dependent attenuation of dexamethasone-induced muscle atrophy in mice.
Many inflammatory and autoimmune diseases are treated using synthetic glucocorticoids. However, excessive glucocorticoid can often cause unpredictable effects including muscle atrophy. Endogenous glucocorticoid levels robustly fluctuate in a circadian manner and peak just before the onset of the active phase in both humans and nocturnal rodents. The present study determines whether muscle atrophy induced by exogenous glucocorticoid can be avoided by optimizing dosing times. We administered single daily doses of the glucocorticoid analog dexamethasone (Dex) to mice for 10 days at the times of day corresponding to peak (early night) or trough (early morning) endogenous glucocorticoid levels. Administration at the acrophase of endogenous glucocorticoids significantly attenuated Dex-induced wasting of the gastrocnemius (Ga) and tibialis anterior (TA) muscles that comprise mostly fast-twitch muscle fibers. Real-time RT-PCR revealed that the Dex-induced mRNA expression of genes encoding the atrophy-related ubiquitin ligases Muscle Atrophy F-box (Fbxo32, also known as MAFbx/Atrogin-1) and Muscle RING finger 1 (Trim63, also known as MuRF1) in the Ga and TA muscles was significantly attenuated by Dex when administered during the early night. Dex negligibly affected the weight of the soleus (So) muscle that mostly comprises slow-twitch muscle fibers, but significantly and similarly decreased the weight of the spleen at both dosing times. These results suggest that glucocorticoid-induced muscle atrophy can be attenuated by optimizing the dosing schedule.


Primary care treatment guidelines for skin infections in Europe: congruence with antimicrobial resistance found in commensal Staphylococcus aureus in the community.
Over 90% of antibiotics for human use in Europe are prescribed in primary care. We assessed the congruence between primary care treatment guidelines for skin infections and commensal Staphylococcus aureus (S. aureus) antimicrobial resistance levels in community-dwelling persons. The prevalence of antimicrobial resistance in S. aureus was analysed by taking nose swabs from healthy primary care patients in nine European countries (total N = 32,032). Primary care treatment guidelines for bacterial skin infections were interpreted with respect to these antimicrobial resistance patterns. First- and second-choice recommendations were assessed and considered congruent if resistance to the antibiotic did not exceed 20%. We included primary care treatment guidelines for impetigo, cellulitis, folliculitis and furuncle. Treatment recommendations in all countries were consistent: most of the first-choice recommendations were beta-lactams, both for children and adults. Antimicrobial resistance levels were low, except for penicillin (on average 73% resistance). Considerable variation in antimicrobial resistance levels was found between countries, with Sweden displaying the lowest levels and Spain the highest. In some countries resistance to penicillin and azithromycin was significantly higher in children (4-17 years) compared with adults. Most of the first- and second-choice recommendations in the treatment guidelines for skin infections were congruent with commensal S. aureus antimicrobial resistance patterns in the community, except for two recommendations for penicillin. Given the variation in antimicrobial resistance levels between countries, age groups and health care settings, national data regarding antimicrobial resistance in the community should be taken into account when updating or developing primary care treatment guidelines.


Low molecular weight heparinoid, ORG 10172 (danaparoid), and outcome after acute ischemic stroke: a randomized controlled trial. The Publications Committee for the Trial of ORG 10172 in Acute Stroke Treatment (TOAST) Investigators.
Anticoagulation with unfractionated heparin is used commonly for treatment of acute ischemic stroke, but its use remains controversial because it has not been shown to be effective or safe. Low molecular weight heparins and heparinoids have been shown to be effective in preventing deep vein thrombosis in persons with stroke, and they might be effective in reducing unfavorable outcomes following ischemic stroke. To test whether an intravenously administered low molecular weight heparinoid, ORG 10172 (danaparoid sodium), increases the likelihood of a favorable outcome at 3 months after acute ischemic stroke. Randomized, double-blind, placebo-controlled, multicenter trial. Between December 22, 1990, and December 6, 1997, 1281 persons with acute stroke were enrolled at 36 centers across the United States. A 7-day course of ORG 10172 or placebo was given initially as a bolus within 24 hours of stroke, followed by continuous infusion in addition to the best medical care. Doses were adjusted in response to anti-factor Xa activity. Favorable outcome rated as the combination of a Glasgow Outcome Scale score of I or II and a modified Barthel Index of 12 or greater on a scale of 0 to 20 at 3 months or 7 days; very favorable outcome was recorded for the combination of a Glasgow Outcome Scale of I and a Barthel Index of 19 or 20 at 3 months or 7 days. At 3 months, 482 (75.2%) of 641 persons assigned to treatment with ORG 10172 and 467 (73.7%) of 634 patients treated with placebo had favorable outcomes (P=.49); 49.5% and 47%, respectively, of patients in each group had very favorable outcomes at 3 months. At 7 days, 376 (59.2%) of 635 persons given ORG 10172 and 344 (54.3%) of 633 receiving placebo had favorable outcomes (P=.07). For the same interval, 215 (33.9%) of 635 persons given ORG 10172 and 176 (27.8%) of 633 persons administered placebo had very favorable outcomes (P=.01; odds ratio, 1.36; 95% confidence interval, 1.06-1.73). Within 10 days of onset of treatment, serious intracranial bleeding events occurred in 14 patients given ORG 10172 (15 events) and in 4 placebo-treated patients (5 events) (P=.05). Despite an apparent positive response to treatment at 7 days, emergent administration of the antithrombotic agent, ORG 10172, is not associated with an improvement in favorable outcome at 3 months.


Rapid and sensitive detection of Bartonella bacilliformis in experimentally infected sand flies by loop-mediated isothermal amplification (LAMP) of the Pap31 gene.
Carrion' disease, caused by Bartonella bacilliformis, remains truly neglected due to its focal geographical nature. A wide spectrum of clinical manifestations, including asymptomatic bacteremia, and lack of a sensitive diagnostic test can potentially lead to a spread of the disease into non-endemic regions where competent sand fly vectors may be present. A reliable test capable of detecting B. bacilliformis is urgently needed. Our objective is to develop a loop-mediated isothermal amplification (LAMP) assay targeting the pap31 gene to detect B. bacilliformis. The sensitivity of the LAMP was evaluated in comparison to qPCR using plasmid DNA containing the target gene and genomic DNA in the absence and presence of human or sand fly DNA. The detection limit of LAMP was 1 to 10 copies/µL, depending on the sample metrics. No cross-reaction was observed when testing against a panel of various closely related bacteria. The utility of the LAMP was further compared to qPCR by the examination of 74 Lutzomyia longipalpis sand flies artificially fed on blood spiked with B. bacilliformis and harvested at days (D) 1, 3, 5, 7 and 9 post feeding. Only 86% of sand flies at D1 and 63% of flies at D3 were positive by qPCR. LAMP was able to detect B. bacilliformis in all those flies confirmed positive by qPCR. However, none of the flies after D3 were positive by either LAMP or qPCR. In addition to demonstrating the sensitivity of the LAMP assay, these results suggest that B. bacilliformis cannot propagate in artificially fed L. longipalpis. The LAMP assay is as sensitive as qPCR for the detection of B. bacilliformis and could be useful to support diagnosis of patients in low-resource settings and also to identify B. bacilliformis in the sand fly vector.


Interface catalytic regulation via electron rearrangement and hydroxyl radicals triggered by oxygen vacancies and heavy metal ions.
Although the enhanced intrinsic activities of some nano-metal oxides are obtained by manufacturing oxygen vacancies (OVs), the effect of multiple roles of OVs is ambiguous. Herein, an interface catalytic regulation via electron rearrangement and hydroxyl radicals (˙OH) was proposed with the designed ZrO2 hollow sphere rich in OVs (Vo-rich ZrO2). Surprisingly, it was shown that the catalytic ability of Vo-rich ZrO2 was 9.9 times higher than that of ZrO2 with little OVs in electrochemical catalytic reduction of Pb(ii). It was found that the generation of Zr2+ and Zr3+ caused by OVs results in the rearrangement of abundant free electrons to facilitate the catalytic reaction rates. The longer bond length between Vo-rich ZrO2 and reactants, and the lower adsorption energy are beneficial for reactants to desorb, improving the conversion rates. Besides, the produced ˙OH were captured which were induced by OVs and trace divalent heavy metal ions in in situ electron paramagnetic resonance (EPR) experiments, contributing to lowering the energy barriers. This study not only revealed the enhanced interface catalytic effect of electron rearrangement and generated ˙OH triggered by OVs, but also provided unique insights into interface catalytic regulation on nano-metal oxides simulated by OVs.


Iron Chelator Transmetalative Approach to Inhibit Human Ribonucleotide Reductase.
Efforts directed at curtailing the bioavailability of intracellular iron could lead to the development of broad-spectrum anticancer drugs given the metal's role in cancer proliferation and metastasis. Human ribonucleotide reductase (RNR), the key enzyme responsible for synthesizing the building blocks of DNA replication and repair, depends on Fe binding at its R2 subunit to activate the catalytic R1 subunit. This work explores an intracellular iron chelator transmetalative approach to inhibit RNR using the titanium(IV) chemical transferrin mimetic (cTfm) compounds Ti(HBED) and Ti(Deferasirox)2. Whole-cell EPR studies reveal that the compounds can effectively attenuate RNR activity though seemingly causing different changes to the labile iron pool that may account for differences in their potency against cells. Studies of Ti(IV) interactions with the adenosine nucleotide family at pH 7.4 reveal strong metal binding and extensive phosphate hydrolysis, which suggest the capacity of the metal to disturb the nucleotide substrate pool of the RNR enzyme. By decreasing intracellular Fe bioavailability and altering the nucleotide substrate pool, the Ti cTfm compounds could inhibit the activity of the R1 and R2 subunits of RNR. The compounds arrest the cell cycle in the S phase, indicating suppressed DNA replication, and induce apoptotic cell death. Cotreatment cell viability studies with cisplatin and Ti(Deferasirox)2 reveal a promising synergism between the compounds that is likely owed to their distinct but complementary effect on DNA replication.


An Approach to Quantify Anisotropic Multiaxial Failure of the Annulus Fibrosus.
Tears in the annulus fibrosus (AF) of the intervertebral disk (IVD) occur due to multiaxial loading on the spine. However, most existing AF failure studies measure uniaxial stress, not the multiaxial stress at failure. Delamination theory, which requires advanced structural knowledge and knowledge about the interactions between the AF fibers and matrix, has historically been used to understand and predict AF failure. Alternatively, a simple method, the Tsai-Hill yield criteria, could describe multiaxial failure of the AF. This yield criteria uses the known tissue fiber orientation and an equation to establish the multiaxial failure stresses that cause failure. This paper presents a method to test the multiaxial failure stress of the AF experimentally and evaluate the potential for the Tsai-Hill model to predict these failure stresses. Porcine AF was cut into a dogbone shape at three distinct angles relative to the primary lamella direction (parallel, transverse, and oblique). Then, each dogbone was pulled to complete rupture. The Cauchy stress in the material's fiber coordinates was calculated. These multiaxial stress parameters were used to optimize the coefficients of the Tsai-Hill yield. The coefficients obtained for the Tsai-Hill model vary by an order of magnitude between the fiber and transverse directions, and these coefficients provide a good description of the AF multiaxial failure stress. These results establish both an experimental approach and the use of the Tsai-Hill model to explain the anisotropic failure behavior of the tissue.


Detecting mitochondrial permeability transition by confocal imaging of intact cells pinocytically loaded with calcein.
When studied in vitro, mitochondrial permeability transition (MPT) is associated with an increase in mitochondrial permeability to solutes up to 1500 Da in mass and a loss of electrical potential difference across the inner mitochondrial membrane (Deltapsimit). The MPT has been implicated as being important in cellular calcium homeostasis, autophagy and cell death via necrosis and apoptosis. Thus, it is important to develop a valid technique for accurate measurement of this phenomenon in intact cells. We developed a procedure for the detection of MPT in intact cells that avoids the disadvantages associated with earlier approaches. In this new technique, unmodified (green-fluorescent) calcein is simultaneously introduced into the cytosol of millions of cells by the process of pinocytic loading and, to identify the position of individual mitochondria and to measure Deltapsimit, the cells are counter-stained with a red-fluorescing potentiometric dye. Using this approach with a variety of cell types, we demonstrate that cytosolic calcein is excluded from normal polarized mitochondria but enters them during MPT. This technique may be valuable in studies investigating the cellular functions of MPT.


Silver-catalyzed decarboxylative fluorination of aliphatic carboxylic acids in aqueous solution.
Although fluorinated compounds have found widespread applications in the chemical and materials industries, general and site-specific C(sp(3))-F bond formations are still a challenging task. We report here that with the catalysis of AgNO(3), various aliphatic carboxylic acids undergo efficient decarboxylative fluorination with SELECTFLUOR(®) reagent in aqueous solution, leading to the synthesis of the corresponding alkyl fluorides in satisfactory yields under mild conditions. This radical fluorination method is not only efficient and general but also chemoselective and functional-group-compatible, thus making it highly practical in the synthesis of fluorinated molecules. A mechanism involvinig Ag(III)-mediated single electron transfer followed by fluorine atom transfer is proposed for this catalytic fluorodecarboxylation.


Carnauba wax nanoparticles enhance strong systemic and mucosal cellular and humoral immune responses to HIV-gp140 antigen.
Induction of humoral responses to HIV at mucosal compartments without inflammation is important for vaccine design. We developed charged wax nanoparticles that efficiently adsorb protein antigens and are internalized by DC in the absence of inflammation. HIV-gp140-adsorbed nanoparticles induced stronger in vitro T-cell proliferation responses than antigen alone. Such responses were greatly enhanced when antigen was co-adsorbed with TLR ligands. Immunogenicity studies in mice showed that intradermal vaccination with HIV-gp140 antigen-adsorbed nanoparticles induced high levels of specific IgG. Importantly, intranasal immunization with HIV-gp140-adsorbed nanoparticles greatly enhanced serum and vaginal IgG and IgA responses. Our results show that HIV-gp140-carrying wax nanoparticles can induce strong cellular/humoral immune responses without inflammation and may be of potential use as effective mucosal adjuvants for HIV vaccine candidates.


Are outcomes after total knee arthroplasty worsening over time? A time-trends study of activity limitation and pain outcomes.
To examine whether function and pain outcomes of patients undergoing primary total knee arthroplasty (TKA) are changing over time. The Mayo Clinic Total Joint Registry provided data for time-trends in preoperative and 2-year post-operative activity limitation and pain in primary TKA patients from 1993-2005. We used chi-square test and analysis for variance, as appropriate. Multivariable-adjusted analyses were done using logistic regression. In a cohort of 7,229 patients who underwent primary TKA during 1993-2005, mean age was 68.4 years (standard deviation (SD), 9.8), mean BMI was 31.1 (SD, 6.0) and 55% were women. Crude estimates showed that preoperative moderate-severe overall limitation were seen in 7.3% fewer patients and preoperative moderate-severe pain in 2.7% more patients in 2002-05, compared to 1992-95 (p < 0.001 for both). At 2-years, crude estimates indicated that compared to 1992-95, moderate-severe post-TKA overall limitation was seen in 4.7% more patients and moderate-severe post-TKA pain in 3.6% more patients in 2002-05, both statistically significant (p ≤ 0.018) and clinically meaningful. In multivariable-adjusted analyses that adjusted for age, sex, anxiety, depression, Deyo-Charlson index, body mass index and preoperative pain/limitation, patients had worse outcomes 2-year post-TKA in 2002-2005 compared to 1993-95 with an odds ratio (95% confidence interval (CI); p-value) of 1.34 (95% CI: 1.02, 1.76, p = 0.037) for moderate-severe activity limitation and 1.79 (95% CI: 1.17, 2.75, p = 0.007) for moderate-severe pain. Patient-reported function and pain outcomes after primary TKA have worsened over the study period 1993-95 to 2002-05. This time-trend is independent of changes in preoperative pain/limitation and certain patient characteristics.


An Electrophilic Carbene-Anchored Silylene-Phosphinidene.
The cyclic alkyl(amino) carbene-anchored silylene-phosphinidene was isolated as L-Si-P(:cAAC-Me) (L=benzamidinate) at room temperature, synthesized from the reduction of L-Si(Cl2 )-P(:cAAC-Me) (1) using two equivalents of KC8 . Compound 1 was prepared by the oxidative addition of a chlorophosphinidene to the benzamidinate substituted silylene center. This is the first molecular example of a silylene-phosphinidene characterized by single-crystal X-ray structural analysis. Moreover, 1 H, 31 P, and also 29 Si NMR spectroscopic data supported the formulation of the products. The theoretical calculations of compound 2 are in good agreement with the experimental results.


Solubilized bone morphogenetic protein (BMP) from mouse osteosarcoma and rat demineralized bone matrix.
A selection of proteins including bone morphogenetic protein (BMP) was extracted in a disaggregated form from Dunn osteosarcoma or rat demineralized bone matrix by 4M guanidine hydrochloride (GuHCl) solution without losing its biological activity. The GuHCl extracts of Dunn osteosarcoma were divied into 4 different fractions by cesium chloride (CsCl) density gradients. Under a dissociative condition, the highest new bone yield was obtained in the low dense top one-third fraction, and BMP acitivity declined with increase in the density of each fraction. No BMP potential was observed in the surface-gel fraction under dissociative conditions. Under an associative condition (low GuHCl concentrations), BMP activity appears in the surface-gel fraction, while under a dissociative condition (high concentrations of GuHCl) BMP appears in the fraction below the surface gel. These facts suggest that under associative conditions, BMP aggregates with other low dense proteins in the surface-gel fraction and that this may be the state of aggregation of BMP in cells and matrix in nature. Present observations support the assumption that BMP is a relatively low density protein and excludes the idea of BMP activity in the collagen molecule, per se. A specific protein, with an apparent molecular weight of 63,000 daltons, is present in all fractions that exhibit BMP activity, and absent in fractions that do not exhibit this activity. BMP is not species-specific; rat BMP induces bone formation in mice. CsCl density-gradient centrifugation is an efficient tool for further purification and isolation of BMP.


A wavelet-based single-view reconstruction approach for cone beam x-ray luminescence tomography imaging.
Single-view x-ray luminescence computed tomography (XLCT) imaging has short data collection time that allows non-invasively and fast resolving the three-dimensional (3-D) distribution of x-ray-excitable nanophosphors within small animal in vivo. However, the single-view reconstruction suffers from a severe ill-posed problem because only one angle data is used in the reconstruction. To alleviate the ill-posedness, in this paper, we propose a wavelet-based reconstruction approach, which is achieved by applying a wavelet transformation to the acquired singe-view measurements. To evaluate the performance of the proposed method, in vivo experiment was performed based on a cone beam XLCT imaging system. The experimental results demonstrate that the proposed method cannot only use the full set of measurements produced by CCD, but also accelerate image reconstruction while preserving the spatial resolution of the reconstruction. Hence, it is suitable for dynamic XLCT imaging study.


[Stomach and duodenal cancers in familial adenomatosis coli].
In the last years the number of patients with familial adenomatosis coli and metachrone carcinoma of the upper gastrointestinal tract is increasing. We describe two patients who, 9 and 15 years after colectomy for adenomatosis coli, developed a duodenal carcinoma. Each patient was treated with a partial duodeno-pancreatectomy. Another patient was resected prophylactically because of a diffuse adenomatosis of the duodenum three years after colectomy. We suggest that each patient with a history of colectomy for adenomatosis coli should have regular follow-ups including endoscopy of the upper gastrointestinal tract.


MicroRNA-31 is required for astrocyte specification.
Previously, we determined microRNA-31 (miR-31) is a noncoding tumor suppressive gene frequently deleted in glioblastoma (GBM); miR-31 suppresses tumor growth, in part, by limiting the activity of NF-κB. Herein, we expand our previous studies by characterizing the role of miR-31 during neural precursor cell (NPC) to astrocyte differentiation. We demonstrate that miR-31 expression and activity is suppressed in NPCs by stem cell factors such as Lin28, c-Myc, SOX2 and Oct4. However, during astrocytogenesis, miR-31 is induced by STAT3 and SMAD1/5/8, which mediate astrocyte differentiation. We determined miR-31 is required for terminal astrocyte differentiation, and that the loss of miR-31 impairs this process and/or prevents astrocyte maturation. We demonstrate that miR-31 promotes astrocyte development, in part, by reducing the levels of Lin28, a stem cell factor implicated in NPC renewal. These data suggest that miR-31 deletions may disrupt astrocyte development and/or homeostasis.


Gastrointestinal Carriage Is a Major Reservoir of Klebsiella pneumoniae Infection in Intensive Care Patients.
Klebsiella pneumoniae is an opportunistic pathogen and leading cause of hospital-associated infections. Intensive care unit (ICU) patients are particularly at risk. Klebsiella pneumoniae is part of the healthy human microbiome, providing a potential reservoir for infection. However, the frequency of gut colonization and its contribution to infections are not well characterized. We conducted a 1-year prospective cohort study in which 498 ICU patients were screened for rectal and throat carriage of K. pneumoniae shortly after admission. Klebsiella pneumoniae isolated from screening swabs and clinical diagnostic samples were characterized using whole genome sequencing and combined with epidemiological data to identify likely transmission events. Klebsiella pneumoniae carriage frequencies were estimated at 6% (95% confidence interval [CI], 3%-8%) among ICU patients admitted direct from the community, and 19% (95% CI, 14%-51%) among those with recent healthcare contact. Gut colonization on admission was significantly associated with subsequent infection (infection risk 16% vs 3%, odds ratio [OR] = 6.9, P < .001), and genome data indicated matching carriage and infection isolates in 80% of isolate pairs. Five likely transmission chains were identified, responsible for 12% of K. pneumoniae infections in ICU. In sum, 49% of K. pneumoniae infections were caused by the patients' own unique strain, and 48% of screened patients with infections were positive for prior colonization. These data confirm K. pneumoniae colonization is a significant risk factor for infection in ICU, and indicate ~50% of K. pneumoniae infections result from patients' own microbiota. Screening for colonization on admission could limit risk of infection in the colonized patient and others.


Constitutive activation of the Janus kinase-STAT pathway in T lymphoma overexpressing the Lck protein tyrosine kinase.
The Lck protein, a Src family tyrosine kinase, plays a critical role in T cell maturation and activation. Dysregulation of Lck expression or Lck kinase activity has been implicated in T cell leukemias from mice to humans, although the mechanism underlying Lck-mediated oncogenesis is still largely unclear. We report here that both DNA binding activities and tyrosine phosphorylation of STAT3 and STAT5, but not STAT1, are constitutively enhanced in the mouse T cell lymphoma LSTRA, which is a well-characterized cell line that overexpresses Lck protein and exhibits high levels of Lck kinase activity. Furthermore, Janus kinase 1 (jak1) and Jak2 protein tyrosine kinases are constantly activated in these cells, as determined by their autophosphorylation in an in vitro kinase assay and increased levels of tyrosine phosphorylation on immunoblots. Therefore, like Src-transformed cells, Lck-overexpressing LSTRA cells also exhibit constitutive activation of distinct Jak and STAT proteins.


Ambiguous Genitalia Associated with an Extremely Rare Syndrome: A Case Report of XLAG Syndrome and Review of the Literature.
X-linked lissencephaly, absent corpus callosum, and epilepsy of neonatal onset with ambiguous genitalia comprises the XLAG syndrome and only 15 cases have been reported in literature. Due to its rarity, the exact clinical course and outcome are not known. Exact associations of this disease are also elusive. Hereby we are reporting this extremely rare entity and we searched the English literature extensively to get consolidated knowledge regarding this entity that would help the readers. Pre-natal radiological work-up can detect these malformations, which should be followed by medical termination, counseling and karyotyping. Till date the longest survival noted was 4 years only.


Sertraline exerts its antitumor functions through both apoptosis and autophagy pathways in acute myeloid leukemia cells.
It has been found that sertraline, a widely used antidepressant drug, possessed antitumor roles in a variety of cancers including liver cancer, colorectal cancer and lymphoma. In this study, we provided evidences that sertraline had potent antiproliferative activity not only in acute myeloid leukemia (AML) cell lines but also in the fresh leukemia cells from AML patients, and could induce cell death through both apoptosis and autophagy pathways. Moreover, we found that inhibiting autophagy pathway could partially attenuate sertraline-induced apoptosis and cell growth inhibition, indicating that sertraline-induced autophagy process could facilitate AML cell apoptosis to some degree. However, blocking apoptosis pathway seemed no obvious effects on sertraline-caused autophagy as well as cell growth inhibition. Our results suggested a potential application value of sertraline in the treatment of AML patients, furnishing some perspectives for novel therapeutic strategies in leukemia.


Phlorizin Mitigates Dextran Sulfate Sodium-Induced Colitis in Mice by Modulating Gut Microbiota and Inhibiting Ferroptosis.
Phlorizin (PHZ) is the main active component of apple peel and presents a potential application value. In the past few years, some reports have suggested that PHZ may have antioxidant and anti-inflammatory effects. Herein, we have attempted to assess the protective effects of PHZ on dextran sodium sulfate (DSS)-induced colitis in mice and to determine the underlying molecular mechanisms. Our results suggested that early intervention with PHZ (20, 40, and 80 mg/kg) significantly reduced the severity of DSS-induced colitis in mice, as presented by a longer colon, improved tight junction protein, decreased disease activity index, and attenuated inflammatory factors. Additionally, early intervention with + (20, 40, and 80 mg/kg) significantly inhibited ferroptosis by decreasing the surrogate ferroptosis marker levels (MDA and Iron Content). Additionally, PHZ (80 mg/kg) increased the diversity of intestinal flora in colitic mice by elevating the levels of beneficial bacteria (Lactobacillaceae and Muribaculaceae) and reducing the levels of harmful bacteria (Lachnospiraceae). This indirectly led to an increase in the amount of short-chain fatty acids. A fecal microbial transplantation (FMT) test was conducted to show that PHZ (80 mg/kg) ameliorated ulcerative colitis (UC) by regulating gut dysbiosis. In conclusion, early intervention with PHZ decreased DSS-induced colitis in mice by preserving their intestinal barrier and regulating their intestinal flora.