Molecular cloning and heterologous expression of a biosynthetic gene cluster for the antitubercular agent D-cycloserine produced by Streptomyces lavendulae.
In the present study, we successfully cloned a 21-kb DNA fragment containing a d-cycloserine (DCS) biosynthetic gene cluster from a DCS-producing Streptomyces lavendulae strain, ATCC 11924. The putative gene cluster consists of 10 open reading frames (ORFs), designated dcsA to dcsJ. This cluster includes two ORFs encoding D-alanyl-D-alanine ligase (dcsI) and a putative membrane protein (dcsJ) as the self-resistance determinants of the producer organism, indicated by our previous work. When the 10 ORFs were introduced into DCS-nonproducing Streptomyces lividans 66 as a heterologous host cell, the transformant acquired DCS productivity. This reveals that the introduced genes are responsible for the biosynthesis of DCS. As anticipated, the disruption of dcsG, seen in the DCS biosynthetic gene cluster, made it possible for the strain ATCC 11924 to lose its DCS production. We here propose the DCS biosynthetic pathway. First, L-serine is O acetylated by a dcsE-encoded enzyme homologous to homoserine O-acetyltransferase. Second, O-acetyl-L-serine accepts hydroxyurea via an O-acetylserine sulfhydrylase homolog (dcsD product) and forms O-ureido-L-serine. The hydroxyurea must be supplied by the catalysis of a dcsB-encoded arginase homolog using the L-arginine derivative, N(G)-hydroxy-L-arginine. The resulting O-ureido-L-serine is then racemized to O-ureido-D-serine by a homolog of diaminopimelate epimerase. Finally, O-ureido-D-serine is cyclized to form DCS with the release of ammonia and carbon dioxide. The cyclization must be done by the dcsG or dcsH product, which belongs to the ATP-grasp fold family of protein.


Tissue plasminogen activator to treat impending pupillary block glaucoma in patients with acute fibrinous HLA-B27 positive iridocyclitis.
To report the use of intracameral tissue plasminogen activator to dissolve fibrinous membranes and break posterior synechiae in patients with acute HLA-B27-positive iridocyclitis with impending pupillary block. Two patients with severe acute fibrinous iridocyclitis and seclusio pupillae were identified. Because of the concern of impending pupillary block, intracameral tissue plasminogen activator (12.5 microg in 0.1 ml, Activase; Genentech, Inc, South San Francisco, California) was injected with a 25-gauge needle through the corneal limbus. Both patients showed complete dissolution of fibrin with disruption of posterior synechiae. There were no adverse events after injection. Neither patient required further invasive intervention, and both fully recovered with medical management. Intracameral tissue plasminogen activator is a safe and effective agent for patients with severe acute iridocyclitis and pupillary seclusion. Patients with clinical signs suggestive of impending pupillary block glaucoma may be considered for tissue plasminogen activator injection to avoid the possible need for emergency surgical iridectomy and synechiolysis.


Late-onset depression: genetic, vascular and clinical contributions.
Neuropsychiatric research needs to examine the relationships between aetiological, genotypic and clinical risk factors and behavioural phenotypes. These relationships can now be examined in older patients with depressive disorders. Key behavioural features, clinical and vascular risk factors and putative genotypes for late-onset neurodegenerative disorders and/or vascular disease were recorded in 78 older patients with depression (mean age = 549 years, S.D. = 14.1) and 22 healthy control subjects (mean age = 55.5 years, S.D. = 9.6). Two or more vascular risks were more common in older patients (65% v. 26% of control subjects, P < 0.01), and in patients with late-onset disorders (82% v. 57% in patients with early-onset disorders, P < 0.05). Patients with late-onset depression had a higher prevalence of the homozygous or heterozygous forms of the C677T mutation of the methylenetetrahydrofolate reductase enzyme (MTHFR)(74% v. 48% in patients with early-onset disorders, P < 0.05). In a multivariate model, only presence of the MTHFR gene mutation predicted late-onset depression (odds ratio = 3.8, 95% CI = 1.1-12.9). Neither apolipoprotein E epsilon 4 or epsilon 2 was associated with depression, late-onset depression, cognitive impairment, or psychomotor change. Patients with apolipoprotein E epsilon 4 were less likely to have psychotic forms of depression. Patients with late-onset depression had an increased rate of the C677T MTHFR gene mutation and other vascular risk factors. This suggests that a proportion of these patients may have genetically-determined and/or other vascular aetiologies. Patients at risk of these disorders may be assisted by currently-available preventative strategies.


Parametric effects in color-difference evaluation.
An experiment was conducted to investigate three parameters affecting color-difference evaluation on a display: 4 sample sizes (2°, 4°, 10°, and 20°), 2 color-difference magnitudes (4 and 8 CIELAB units), and 2 separations (inclusion or exclusion of the separation line between two colors in a pair). Sample pairs surrounding 5 CIE recommended color centers were prepared. In total, 1120 sample pairs of colors were assessed 20 times using the grey-scale method. The experimental results were used to reveal various parametric effects and to verify the performance of different color matching functions (CMFs) and four color difference formulae and uniform color spaces. It was found that there was little difference in terms of ΔE values calculated using different CMFs for all the color models tested. A parametric formula was proposed to predict three parametric effects for sample pairs having no-separation line: 1) differences in sample size, 2) media (surface and self-luminous colors), and 3) color-difference magnitudes.


Mesogenicity drives fractionation in lyotropic aqueous suspensions of multiwall carbon nanotubes.
We describe a simple method for separating carbon nanotubes on the basis of their mesogenicity by fractionating biphasic aqueous suspensions within the Flory chimney of the lyotropic phase diagram. Macroscopic phase separation occurs on centrifuging the biphasic nanotube suspension or allowing it to stand. Long, straight nanotubes with higher mesogenicity (liquid crystalline forming ability) segregate preferentially to the liquid crystalline phase, whereas shorter nanotubes and impurities with lower mesogenicity segregate preferentially to the isotropic phase.


Total, anti-viral, and anti-myelin IgG subclass reactivity in inflammatory diseases of the central nervous system.
Total IgG subclass levels, anti-viral, anti-myelin basic protein (anti-MBP), and anti-ganglioside 1 (anti-GM1) IgG subclass levels were measured in 6 patients with herpes simplex virus encephalitis (HSVE), 16 with borreliosis, 8 with other bacterial infections, 12 with multiple sclerosis (MS), 13 with subacute sclerosing panencephalitis (SSPE), 5 with glioblastoma and 12 controls. Total IgG1 levels were elevated in cerebrospinal fluid (CSF) from all patient groups (but not in the controls), IgG2 in bacterial infections, IgG3 in HSVE and borreliosis and IgG4 in some SSPE patients. The anti-viral (anti-measles, varicella zoster virus and rubella) IgG antibodies in MS were restricted to IgG1, anti-measles IgG to IgG1 and sometimes IgG4 in SSPE, anti-borrelia IgG to IgG1, IgG2 and IgG3. In contrast to anti-viral antibodies, anti-MBP and GM1 antibodies belonged to IgG1, IgG3 or IgG4 in MS. The nature of the immunological activation appears to be reflected in the subclass patterns elicited in the central nervous system. Different IgG subclass patterns in infectious diseases and MS suggest a difference between antigen-specific and non-specific B-cell activation.


An alternative pancreatic anastomosis following pancreaticoduodenectomy.
Pancreatic head resection and reconstruction is technically challenging. Eight patients underwent pancreaticoduodenectomy for either ductal adenocarcinoma (n = 7) or neuroendocrine tumor (n = 1) in the head of the pancreas with a dilated pancreatic duct. The pancreatic stump could not be mobilized to form a standard pancreaticogastrostomy or a pancreaticojejunostomy following resection because of a complete fixation to the splenic vein (n = 2), common hepatic artery (n = 1), or mesentery (n = 3) or inadequate length of the pancreatic remnant (n = 2). After laying open the pancreatic duct along the pancreatic transection margin in the ventral aspect of the pancreas, a longitudinal ventral pancreaticojejunostomy was performed using polydioxanone 3/0 sutures. The average time taken to create this pancreatic anastomosis was less than 10 minutes. This longitudinal ventral pancreatic anastomosis is quick, easy to perform, and a safe alternative method for pancreatic reconstruction after pancreaticoduodenectomy.


Wound Closure and Simultaneous Soft Tissue Augmentation at Fresh Extraction Sockets: A Simplified Technique Adopting a Xenogeneic Collagen Matrix.
A simplified surgical technique is presented for closure and soft tissue augmentation of fresh extraction sockets that utilizes a xenogeneic collagen matrix as a substitute for a combined onlay-interpositional connective tissue graft. Ten alveoli receiving a socket preservation procedure (n = 5) or an immediate submerged implant (n = 5) were sealed with a xenogeneic collagen matrix, stabilized under small palatal and buccal full-thickness envelope flaps. Eight weeks after surgery, full wound closure was achieved in 9 out of 10 sites with satisfactory esthetic integration (mean ΔE score: 3.76). Results from this descriptive observational study suggest that this technique may represent a valuable and minimally invasive solution to simplify closure and soft tissue augmentation of fresh extraction sockets.


A colorimetric determination for glycosidic and bile salt-based detergents: applications in membrane protein research.
Detergents are crucial to the isolation of integral membrane proteins. During membrane protein purification, it is useful to accurately quantify detergent, especially if concentration steps have been used. Previously, this has been difficult and time-consuming. We present a simple, rapid, and sensitive procedure for the quantification of glycosidic and bile salt-based detergents such as dodecylmaltoside, octylglucoside, and CHAPS. The method directly quantifies sugar or cholate moieties via colorimetric reactions with phenol and sulfuric acid. A number of detergents have been screened, and the assay has been validated in the presence of commonly used reagents. In addition to determining the overall detergent concentration in solution, the procedure allows accurate quantification of specific binding of glycosidic or bile salt-based detergents to purified membrane proteins. Both the colorimetric method and the radiometric 14C method were used to determine detergent binding to two integral membrane proteins: the cytochrome cbb3 oxidase from Pseudomonas stutzeri and the turkey beta-adrenergic receptor. Both methods gave similar results. After separating monomeric glycosidic detergent from micellar solutions by ultrafiltration, we used the colorimetric method to determine the concentration of monomeric detergent present. We observed that values obtained are in close agreement with previously determined critical micelle concentrations.


Standardization of ceruloplasmin measurements is still an issue despite the availability of a common reference material.
The purpose of measurement standardization is to achieve closer comparability of results obtained using different commercial systems. Regarding serum protein immunoassays, a reference preparation (BCR-470) was released in 1993 and adopted by manufacturers across the world to value-assign their assay calibrators for routine methods to reduce method-dependent variation. Moving from nephelometric (Beckman Immage 800) to turbidimetric determination (Roche Cobas c 501) of seven serum proteins, we preliminarily checked the comparability of results between the two systems. The study was performed according to the CLSI EP9-A protocol on 30 fresh sera, tested on each system in duplicate, and subdivided on two different days, without recalibration and using manufacturers' control materials to validate the runs. Both manufacturers' package inserts provide statements that kit calibrators are traceable to BCR-470. Suggested reference intervals are also the same. Although a fairly good correlation was observed (r = 0.955), the comparison of ceruloplasmin methods produced evidence of highly significant proportional (regression slope, 0.572) and constant bias (intercept, 0.05 g/L). Absolute and percentage mean differences were -0.11 g/L (95% confidence interval (CI) -0.13 to -0.10 g/L) and -39.1% (CI -43.1 to -35.2%), respectively. No other evaluated proteins showed similar problems. Lacking a ceruloplasmin reference method, it is impossible to demonstrate that one of the two assays produces true ceruloplasmin values. The problem is, however, that results coming from the two assays are clearly not comparable. This may be either due to a lack of commutability of the reference material with biological samples in the evaluated assays or to calibration problems by manufacturers in one of the stages of the calibration hierarchy.


Improved Lubricating Performance by Combining Oil-Soluble Hairy Silica Nanoparticles and an Ionic Liquid as an Additive for a Synthetic Base Oil.
This article reports on improved lubricating performance by combining oil-soluble poly(lauryl methacrylate) brush-grafted silica nanoparticles (hairy NPs or HNP) and an oil-miscible phosphonium-phosphate ionic liquid (IL) as a friction-reducing additive for a polyalphaolefin (PAO) oil. The HNP was synthesized by surface-initiated reversible addition-fragmentation chain transfer polymerization. At a total concentration of 2% and sufficiently high individual concentrations for HNP and IL in PAO, high-contact stress, ball-on-flat reciprocating tribological tests showed that the friction decreased by up to 23% compared with 2% HNP alone in PAO and by up to 35% compared to the PAO mixed with 2% IL. Scanning electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray photoelectron spectroscopy (XPS) analysis revealed that the tribofilm formed from the PAO containing 1% HNP + 1% IL was enriched with both Si and P, indicating that both hairy NPs and IL were involved in the tribochemical reactions. In addition, the O 1s and Si 2p peaks in the core-level XPS spectra exhibited significant shifts for the mixture of 1% HNP + 1% IL compared to those for 2% HNP, suggesting the possible formation of new covalent bonds. These results indicated that HNP and IL reacted with each other and also with the metal substrate during the rubbing process, which likely strengthened the tribofilm and its bonding with the substrate and thus further improved the lubrication.


Mapping ex ante risks of COVID-19 in Indonesia using a Bayesian geostatistical model on airport network data.
A rapid response to global infectious disease outbreaks is crucial to protect public health. Ex ante information on the spatial probability distribution of early infections can guide governments to better target protection efforts. We propose a two-stage statistical approach to spatially map the ex ante importation risk of COVID-19 and its uncertainty across Indonesia based on a minimal set of routinely available input data related to the Indonesian flight network, traffic and population data, and geographical information. In a first step, we use a generalised additive model to predict the ex ante COVID-19 risk for 78 domestic Indonesian airports based on data from a global model on the disease spread and covariates associated with Indonesian airport network flight data prior to the global COVID-19 outbreak. In a second step, we apply a Bayesian geostatistical model to propagate the estimated COVID-19 risk from the airports to all of Indonesia using freely available spatial covariates including traffic density, population and two spatial distance metrics. The results of our analysis are illustrated using exceedance probability surface maps, which provide policy-relevant information accounting for the uncertainty of the estimates on the location of areas at risk and those that might require further data collection.


Computed tomography and magnetic resonance appearance of sporadic meningioangiomatosis correlated with pathological findings.
Meningioangiomatosis (MA) is a rare benign localized lesion of leptomeninges and underlying cerebral cortex. Preoperative diagnosis is difficult and challenging because of its diverse clinical, pathological, and imaging features. We retrospectively analyzed 7 cases of MA to explore their imaging features and correlate with pathological findings. Imaging studies including computed tomography (CT) and magnetic resonance imaging (MRI) were retrospectively reviewed in 7 patients with surgically and pathologically verified intracranial MA (not associated with neurofibromatosis). Computed tomography studies were performed in axial plane without iodinated contrast-material administration; magnetic resonance studies consisted of axial T1-weighted, T2-weighted, Fluid attenuated Inversion Recovery (FLAIR), and postcontrast T1-weighted sequences and coronal or sagittal precontrast and postcontrast T1-weighted sequences. Computed tomography showed focal extensively calcified lesions in 3 cases, lesions with patchy calcification in 2 cases, and no apparent calcification in 2 cases. Magnetic resonance imaging demonstrated predominantly hypointensity on T1-weighted images and hyperintensity on T2-weighted images. Six of 7 cases exhibited gyriform hyperintensity on FLAIR sequences, which correlated with proliferating microvessels with perivascular cuffs of spindle-cell proliferation within the cortex on histopathological analysis. After contrast-material administration, all but 1 showed heterogeneous enhancement. The nonenhancing lesion on MRI was completely calcified on CT. Gyriform hyperintensity on FLAIR sequence is the main MRI feature of MA, which correlates with proliferating microvessels with perivascular cuffs of spindle-cell proliferation within the cortex on pathological analysis. Plain CT scan is essential to demonstrate the extent of calcification of these lesions.


Skin, Liver, and Kidney Interactions Contribute to Skin Dryness in Aging KK-Ay/Tajcl Mice.
Type 2 diabetes is a lifestyle-related disease that affects people worldwide and is especially prevalent in the elderly. Many elderly people with diabetes also complain of dry skin; however, the relationship between aging and dry skin in type 2 diabetes is unknown. The purpose of this study was to examine the interaction between aging and dry skin using the specific pathogen-free KK-Ay/TaJcl type 2 diabetes mouse model. Skin dryness in this model increases with age and was evaluated at 10, 27, 40, and 50 weeks. We observed increased mast cell expression, increased histamine and matrix metalloproteinase-1 levels, and decreased collagen expression in the skin of aging KK-Ay/TaJcl mice. In addition, the increased expression of angiopoietin 2, interleukin-6, tumor necrosis factor-α, and endostatin in the blood indicated kidney damage in this model. Aging KK-Ay/TaJcl mice also showed fatty liver pathology, which led to increased reactive oxygen species in the blood and liver, as well as the increased expression of M1 macrophages in the liver. These results showed that dry skin is associated with skin, kidney, and liver interactions in an aging type 2 diabetes mouse model.


[Nutritional status, food consumption and physical activity in female school children of different socioeconomic levels from Santiago, Chile].
A high prevalence of obesity is the main public health problem in Chilean school children. To compare the nutritional status, consumption of selected foods and extracurricular physical activity (PA) habits in school children of different socioeconomic levels as a baseline for developing effective educational interventions. Cross-sectional study that determined the body mass index, food consumption and physical activity with previously validated instruments in 202 and 358 girls from 3rd to 8th grade in schools of medium-high and low socioeconomic level (SEL) from Santiago, Chile, respectively. Compared to their counterparts of low socioeconomic level (SEL), the prevalence of obesity was significantly lower in 8-9 year-old girls of medium high SEL (19% and 9%, respectively, p =0.012) and 12-13 year-old (12% and 2.5% respectively, p =0.008). Also median daily intake of dairy products was higher in girls of medium high SEL (250 and 470 ml/day, respectively). The intake of fruits and vegetables was similar (200 g/d); and the intake of bread was lower (230 and 70 g/day, respectively, p <0.01). Consumption of energy-dense foods was lower in 10-13 year-old girls of medium high SEL (80 and 50 g/day, respectively, p <0.01). 45% of 8-9 year-old girls and 35% of 12-13 year-old girls of both SEL engaged in PA four or more times per week (NS). Although the prevalence of obesity in girls of medium-high SEL was not as high as in those from low SEL, it is still high. There is a need for educational interventions to improve their food and PA habits and to promote an environment that enhances healthy behaviors.


Paradoxical phosphorylation of the serine 199 on tau proteins from young individuals.
The microtubule-associated tau proteins are abnormally aggregated in many tauopathies. Phosphorylation modulates the functions of tau. The serine 199 residue of tau is abnormally phosphorylated at early and late stages of Alzheimer's disease. The presence of the phosphorylated Ser199 was investigated in autopsy-derived and biopsy-derived brain tissue samples from non-demented individuals. A paradoxical expression was found in the hippocampus of the youngest ones, in granule cells of the dentate gyrus and in pyramidal cells of the Ammon's horn, which are particularly prone to neurodegeneration in several tauopathies. The rate of positive cells decreased with age. These data emphasize the importance of the phosphorylation of the Ser199 residue of tau in ageing and susceptibility to neurodegeneration.


Possible role of low dose dexamethasone administration in listeria monocytogenes meningoencephalitis: A case series.
Listeria (L.) monocytogenes is a gram-positive, non-sporulating, facultatively anaerobic bacillus transmitted to humans through ingestion of contaminated foods. Listeriosis represents the third most common cause of death from foodborne illness, with a mortality rate of 20-30%, especially for patients affected by an invasive disease, which typically affects immunocompromised patients, pregnant women, the elderly, and neonates. It causes several clinical syndromes, of which meningitis, meningoencephalitis, and sepsis are the most challenging to deal with. Here, five cases of L. monocytogenes meningitis/meningoencephalitis affecting two previously healthy immunocompetent and three immunocompromised adult patients treated with ampicillin plus gentamicin are reported. In addition, all the patients described in this report received a low dose of intravenous dexamethasone; four of them made a full recovery. Additionally, a literature search was performed to better explain the appropriate clinical and therapeutic management approaches for these patients, highlighting the value of dexamethasone administration as part of the therapy.


Th2-M2 immunity in lesions of muscular sarcoidosis and macrophagic myofasciitis.
To analyse the paradox of a lack of giant cell formation and fibrosis in chronic lesions of macrophagic myofasciitis (MMF) in comparison with muscular sarcoidosis (MuS). Inflammatory lesions and contiguous muscle regions from biopsy samples of 10 patients with MuS and 10 patients with MMF were cut out by laser microdissection. Mediators of the T helper cell (Th)1 inducing classical macrophage activation (e.g. STAT1, IFNγ and CXCR3), and Th2 inducing alternative activation of macrophages (e.g. CD206/MRC1, STAT6, SOCS1), molecules involved in development of fibrosis (e.g. TGFβ) and giant cells (e.g. TYROBP), were assessed by immunohistochemistry and real-time polymerase chain reaction (PCR). STAT6-induced Th2 immunity was associated with up-regulated gene expression of MRC1, SOCS1 and TGFB in inflammatory foci, in comparison with adjacent tissue. TYROBP and TREM2, genes regulating giant cell formation, were more strongly expressed in lesions of MuS patients than in those of MMF. TGFβ co-localized with CD206(+) macrophages in MuS but not in MMF. Conversely, Th1 immunity was illustrated by STAT1 staining both in macrophages and myofibres in MuS, but not in MMF. Also, STAT1-induced IFNG and CXCR3 expression in lesions and the surrounding tissue was elevated compared with normal controls, but without statistically significant differences. Giant cell and typical granuloma formations, including fibrogenesis, is dependent on two main mechanisms, both involving specific macrophage activation: a strong Th2-M2 polarization and a significant expression of TYROBP and TGFβ in macrophages. The low-grade alternative activation of macrophages in MMF lesions and poor TYROBP and TGFβco-expression are obviously insufficient to produce giant cells.


Continuous genomic diversification of long polynucleotide fragments drives the emergence of new SARS-CoV-2 variants of concern.
Highly transmissible or immuno-evasive SARS-CoV-2 variants have intermittently emerged, resulting in repeated COVID-19 surges. With over 6 million SARS-CoV-2 genomes sequenced, there is unprecedented data to decipher the evolution of fitter SARS-CoV-2 variants. Much attention has been directed to studying the functional importance of specific mutations in the Spike protein, but there is limited knowledge of genomic signatures shared by dominant variants. Here, we introduce a method to quantify the genome-wide distinctiveness of polynucleotide fragments (3- to 240-mers) that constitute SARS-CoV-2 sequences. Compared to standard phylogenetic metrics and mutational load, the new metric provides improved separation between Variants of Concern (VOCs; Reference = 89, IQR: 65-108; Alpha = 166, IQR: 149-181; Beta 131, IQR: 114-149; Gamma = 164, IQR: 150-178; Delta = 235, IQR: 217-255; and Omicron = 459, IQR: 395-521). Omicron's high genomic distinctiveness may confer an advantage over prior VOCs and the recently emerged and highly mutated B.1.640.2 (IHU) lineage. Evaluation of 883 lineages highlights that genomic distinctiveness has increased over time (R 2 = 0.37) and that VOCs score significantly higher than contemporary non-VOC lineages, with Omicron among the most distinctive lineages observed. This study demonstrates the value of characterizing SARS-CoV-2 variants by genome-wide polynucleotide distinctiveness and emphasizes the need to go beyond a narrow set of mutations at known sites on the Spike protein. The consistently higher distinctiveness of each emerging VOC compared to prior VOCs suggests that monitoring of genomic distinctiveness would facilitate rapid assessment of viral fitness.


β-Cyclodextrin-based polycationic amphiphilic "click" clusters: effect of structural modifications in their DNA complexing and delivery properties.
Monodisperse facial amphiphiles consisting of a β-cyclodextrin (βCD) platform exposing a multivalent display of cationic groups at the primary rim and bearing hydrophobic chains at the secondary oxygens have been prepared by implementing two very robust "click" methodologies, namely cuprous cation-catalyzed azide-alkyne cycloaddition (CuAAC) and thiourea-forming reaction. Most interestingly, the use of solid-supported Cu(I) catalysts was found to be very well suited for multiple CuAAC while facilitating purification of the C(7)-symmetric macromolecular triazole adducts. The strategy is compatible with molecular diversity-oriented approaches, which has been exploited to generate a small library of click polycationic amphiphilic CDs (paCDs) for assessing the influence of structural modifications in the ability to complex, compact, and protect pDNA and the efficiency of the resulting paCD:pDNA nanocomplexes (CDplexes) to deliver DNA into cells and promote transfection. The results indicate that fine-tuning the hydrophilic/hydrophobic balance is critical to achieve optimal self-assembling properties and stability of the resulting CDplexes in saline- and serum-containing media. Triazole-type paCDs were, in general, less efficient in promoting gene transfection than thiourea-type derivatives. Nevertheless, the current body of results support that the "dual click" approach implying sequential CuAAC and thiourea-forming reactions represents a versatile strategy to optimize the gene delivery capabilities of cyclodextrin-based facial amphiphiles.


A mutation in yeast topoisomerase II that confers hypersensitivity to multiple classes of topoisomerase II poisons.
A mutation was constructed in the CAP homology domain of yeast topoisomerase II that resulted in hypersensitivity to the intercalating agent N-[4-(9-acridinylamino)-3-methoxy-phenyl]methanesulfonamide and the fluoroquinolone 6, 8-difluoro-7-(4'-hydroxyphenyl)-1-cyclopropyl-4-quinolone-3-carboxyli c acid, but not to etoposide. This mutation, which changes threonine at position 744 to proline, also confers hypersensitivity to anti-bacterial fluoroquinolones. The purified T744P mutant protein had wild type enzymatic activity in the absence of drugs, and no alteration in drug-independent DNA cleavage. Enhanced DNA cleavage in the presence of N-[4-(9-acridinylamino)-3-methoxy-phenyl]methanesulfonamide and fluoroquinolones was observed, in agreement with the results observed in vivo. DNA cleavage was also seen in the presence of norfloxacin and oxolinic acid, two quinolones that are inactive against eukaryotic topoisomerase II. The hypersensitivity was not associated with heat-stable covalent complexes, as was seen in another drug-hypersensitive mutant. Molecular modeling suggests that the mutation in the CAP homology domain may displace amino acids that play important roles in catalysis by topoisomerase II and may explain the drug-hypersensitive phenotype.


Genetic variants on chromosome 5p12 are associated with risk of breast cancer in African American women: the Black Women's Health Study.
Two single nucleotide polymorphisms (SNPs), rs4415084, and rs10941679 on chromosome 5p12 were associated with risk of breast cancer in a recent genome-wide association study (GWAS) of women of European ancestry. Both SNPs are located in a large high-LD region and the causal variant(s) are still unknown. We conducted a nested case-control study in a cohort of African American women to replicate and narrow the region carrying the causal variant(s). We evaluated 14 tagging SNPs in a 98 kb LD block surrounding the index SNPs in 886 breast cancer cases and 1,089 controls from the Black Women's Health Study. We used the Cochran-Armitage trend test to assess association with breast cancer risk. Odds ratios were derived from logistic regression analyses adjusted for potential confounders including percent European admixture. We confirmed the reported association of rs4415084 SNP with overall risk of breast cancer (P = 0.06), and, as in the original study, observed a stronger association with estrogen receptor positive tumors (P = 0.03). We identified four other SNPs (rs6451770, rs12515012, rs13156930, and rs16901937) associated with risk of breast cancer at the nominal alpha value of 0.05; all of them were located in a 59 kb HapMap YRI LD block. After correction for multiple testing, the association with SNP rs16901937 remained significant (P permutated = 0.038). The G allele was associated with a 21% increased risk of breast cancer overall and with a 32% increase in tumors positive for both estrogen and progesterone receptors. The present results from an African ancestry (AA) population confirm the presence of breast cancer susceptibility genetic variants in the chromosome 5p12 region. We successfully used the shorter range of LD in our AA sample to refine the localization of the putative causal variant.


Swimming motility in a longitudinal collection of clinical isolates of Burkholderia cepacia complex bacteria from people with cystic fibrosis.
Chronic bacterial lung infections in cystic fibrosis (CF) are the leading cause of morbidity and mortality. While a range of bacteria are known to be capable of establishing residence in the CF lung, only a small number have a clearly established link to deteriorating clinical status. The two bacteria with the clearest roles in CF lung disease are Pseudomonas aeruginosa and bacteria belonging to the Burkholderia cepacia complex (BCC). A number of common adaptations by P. aeruginosa strains to chronic lung infection in CF have been well described. Typically, initial isolates of P. aeruginosa are nonmucoid and display a range of putative virulence determinants. Upon establishment of chronic infection, subsequent isolates ultimately show a reduction in putative virulence determinants, including swimming motility, along with an acquisition of the mucoid phenotype and increased levels of antimicrobial resistance. Infections by BCC are marked by an unpredictable, but typically worse, clinical outcome. However, in contrast to P. aeruginosa infections in CF, studies describing adaptive changes in BCC bacterial phenotype during chronic lung infections are far more limited. To further enhance our understanding of chronic lung infections by BCC bacteria in CF, we assessed the swimming motility phenotype in 551 isolates of BCC bacteria from cystic fibrosis (CF) lung infections between 1981 and 2007. These data suggest that swimming motility is not typically lost by BCC during chronic infection, unlike as seen in P. aeruginosa infections. Furthermore, while we observed a statistically significant link between mucoidy and motility, we did not detect any link between motility phenotype and clinical outcome. These studies highlight the need for further work to understand the adaptive changes of BCC bacteria during chronic infection in the CF lung.


Common birth defects in South African Blacks.
The prevalence rates of some of the common birth defects have been ascertained in a Black population in South Africa. The births of 29,633 infants were studied retrospectively from hospital records. The commonest defect was found to be polydactyly (10,4/1,000 births), and the other defects investigated were talipes equinovarus (1,55/1000 births), hydrocephalus (1,3/1,000 births), spina bifida and anencephaly (0,78 and 0,40/1,000 births respectively) and facial clefts (0,30/1,000 births). These rates were compared with those found in other population groups in South Africa and, in some cases, in other African countries and in Negro groups in the USA and Canada.


Polyethylene glycol-based ultrasound-assisted extraction of magnolol and honokiol from Cortex Magnoliae Officinalis.
In this study, a kind of green solvent named polyethylene glycol (PEG) was developed for the ultrasound-assisted extraction (UAE) of magnolol and honokiol from Cortex Magnoliae Officinalis. The effects of PEG molecular weight, PEG concentration, sample size, pH, ultrasonic power and extraction time on the extraction of magnolol and honokiol were investigated to optimise the extraction conditions. Under the optimal extraction conditions, the PEG-based UAE supplied higher extraction efficiencies of magnolol and honokiol than the ethanol-based UAE and traditional ethanol-reflux extraction. Furthermore, the correlation coefficient (R(2)), repeatability (relative standard deviation, n = 6) and recovery confirmed the validation of the proposed extraction method, which were 0.9993-0.9996, 3.1-4.6% and 92.3-106.8%, respectively.


Use of tilapia piscidin 3 (TP3) to protect against MRSA infection in mice with skin injuries.
Antimicrobial peptides (AMPs), represent promising agents for new therapeutic approaches of infected wound treatment, on account of their antimicrobial and wound closure activities, and low potential for inducing resistance. However, therapeutic applications of these AMPs are limited by their toxicity and low stability in vivo. Previously, we reported that the 23 amino-acid designer peptide TP3 possessed antimicrobial activities. Here, we analyzed the wound closure activities of TP3 both and in vivo. TP3 at doses of up to 40 μg/ml did not affect the viability of baby hamster kidney cells. Furthermore, TP3 was found to be highly effective at combating peritonitis and wound infection caused by MRSA in mouse models, without inducing adverse behavioral effects or liver or kidney toxicity. TP3 treatment increased survival by 100% at 8 days after infection, and accelerated the progression of proliferation, remodeling, and maturation of infected wounds. Taken together, our results indicate that TP3 enhances the rate of survival of mice infected with the bacterial pathogen MRSA through both antimicrobial and immunomodulatory effects. Overall, these results suggest that TP3 may be suitable for development as a novel topical agent for treatment of infected wounds.


Long non-coding RNA maternally expressed 3 (MEG3) regulates isoflurane-induced cognitive dysfunction by targeting miR-7-5p.
This study aimed to investigate the role and mechanism of long non-coding RNA maternally expressed gene 3 (MEG3) in cognitive dysfunction induced by isoflurane (ISO). Morrier water maze analysis was performed to evaluate the cognitive function of rats. Modified modified neurological severity score (mNSS) scores were assessed for neurological damage. The levels of MEG3 in hippocampal tissues of rats and hippocampal neuron cell lines HT22 were examined by reverse transcription-quantitative polymerase chain reaction (qRT-PCR). Moreover, the cell viability and apoptosis were assessed by the Cell Counting Kit-8 (CCK-8) and flow cytometry assay. Indicators of inflammation and oxidative stress were determined using enzyme-linked immunosorbent assay (ELISA) and commercial assay kits. Relationship between MEG3 and microRNA (miR)-7-5p was verified by the dual-luciferase reporter gene assay. MEG3 was increased in hippocampal tissues and HT22 after ISO treatment (p < 0.05). MEG3 downregulation alleviated the increase in neurological severity score and cognitive dysfunction caused by ISO treatment (p < 0.05). In vitro, MEG3 downregulation alleviates the decrease in cell activity and increased apoptosis induced by ISO. What's more, MEG3 reduction eliminated activation of neuroinflammation and oxidative stress promoted by ISO treatment in rats and HT22 (p < 0.05). MEG3 was confirmed to specifically bind to miR-7-5p. Inhibition of miR-7-5p eliminated the alleviating effects of MEG3 downregulation on cognitive dysfunction caused by ISO treatment. Decreased MEG3 alleviates cognitive dysfunction caused by ISO by targeting miR-7-5p and play a neuroprotective effect. We present a strategy for MEG3 as a potential target for brain protection during anesthesia.


[Treatment of primary osteoporosis in men].
Primary osteoporosis in men occurs less frequently and thus tends to be less noticed than that in women. However, once a femoral neck fracture occurs in men, patients are more likely to be disabled than women. Thus, implementing treatments of osteoporosis before the occurrence of femoral neck fractures is important in terms of men's quality of life. Several studies have shown that bisphosphonate is a mainstay of the treatment of the disorder, by increasing bone mineral density and reducing the risk of vertebral fractures.


Anti-streptolysin O titers in normal healthy children of 5-15 years.
Antistreptolysin O (ASO) levels vary with age group of the study population and geographical locations. The present study was undertaken to determine the upper limit of normal of ASO in 200 normal children of 5-15 years of age with no history of recent sore throat infection. The standard tube dilution method (WHO) was used for estimating ASO titers. It was found that 239 IU was the upper limit of normal in the study population, which can be considered as the baseline ASO titer. This can provide useful guidelines for physicians in the interpretation of elevated ASO titers in cases of suspected acute rheumatic fever.


[Non-heme iron proteins as an alternative system of antioxidant defense in the cells of strictly anaerobic microorganisms: a review].
Enzymatic systems accounting for the relative oxygen resistance of multiple strict anaerobes are reviewed, with emphasis on molecular-biological properties and action mechanisms of non-heme iron proteins (neelaredoxins, desulfoferrodoxins, and rubrerythrins). These unique proteins, which are widespread in anaerobes, comprise a system of antioxidant defense against toxic effects of oxygen and products of its incomplete reduction (an alternative to the classic antioxidant system involving superoxide dismutase and catalase). The role of the superoxide reductase-mediated elimination of endogenous superoxide radicals is discussed. This extremely efficient means of rapid superoxide radical detoxification underlies the preferred mechanism for maintaining the optimum balance between oxidized and reduced forms of some proteins in the cells of strict anaerobes.


Stigmatization of anorexia nervosa.
The current study examined the perceptions about an individual with anorexia nervosa (AN) relative to perceptions about a healthy person and a person with another mental or nonmental illness. Ninety-one participants recruited from the community completed questionnaires targeting perceptions about 4 individuals: a healthy person, a person with asthma, a person with schizophrenia, and a person with AN. Evaluations of personal characteristics were most negative for persons with AN. Participants believed the person with AN was most to blame for his/her condition, was best able to pull him/herself together if he/she wanted to, and was most acting this way for attention and that biological factors were least relevant in developing the illness. Negative perceptions of a person with AN fell into stigma categories of self-attribution and responsibility. These attitudes may contribute to reluctance to seek treatment among individuals with AN.


[E-Cigarettes on Everyone's Lips - Current Representative Data on Use Among Adolescents and Adults].
Background International data suggest an increase in e-cigarette use. However, there are still significant deficits in the reporting of its current prevalence. The present study reports representative data on e-cigarette use that make it possible to differentiate users both by the nicotine content of the e-cigarette liquids being used and by their smoking status. Methods and material The underlying data are taken from the National Cancer Aid Monitoring (NCAM). In its first wave, 3000 persons aged between 14 and 45 were interviewed by telephone. Point estimates, confidence intervals and determinants of use will be presented. Results Overall, 20 % of respondents had used an e-cigarette before, of which 80 % used liquids containing nicotine. Men, younger persons, persons with a migrant background and persons with a lower education level were more likely to have used an e-cigarette before than their respective reference groups. Half of all ever users of e-cigarettes also smoked tobacco cigarettes, while roughly one third had never used tobacco products before. Conclusions For the first time, a representative study was conducted combining questions on the use of e-cigarettes with questions on smoking tobacco cigarettes and the nicotine content of the liquids being used. Target groups for future prevention measures are adolescents, as they had the highest prevalence of e-cigarette use, as well as non-smokers, since one third of e-cigarette users had never used a tobacco product before. Both groups also seem to be relevant target groups for which the introduction to e-cigarettes should be prevented.


Exploring subconscious bias.
Implicit biases may lead to subconscious evaluations of a person based on irrelevant characteristics such as race or gender. This audit investigates the presence of implicit bias in the management of patients who missed appointments in our department. This study retrospectively analysed discharge rates in 285 patients who missed an out-patient appointment between 1 May 2020 and 1 April 2021 at Guy's and St Thomas' Hospital. After reading the patients' names, 285 patients were categorised into genders, and ethnic categories of: White British; Black, Asian and minority (non-White) ethnic ('BAME'); and other White. There were no differences in discharge rates in terms of self-reported ethnic and gender groups. However, patients perceived as White British were less likely to be discharged when compared to patients perceived as Black, Asian and minority ethnic (35 per cent vs 58 per cent). Discharge rates for perceived gender did not differ. Implicit bias may influence decision-making regarding whether to rebook a patient after missing an appointment.


β-sitosterol inhibits high cholesterol-induced platelet β-amyloid release.
Recently, increasing evidence has linked high cholesterol to the pathogenesis of Alzheimer's disease (AD), suggesting that cholesterol may be a target for developing new compounds to prevent or treat AD. Plant sterols, a group of sterols enriched in plant oils, nuts, and avocados, have the structure very similar to that of cholesterol, and have been widely used to reduce blood cholesterol. Due to their cholesterol-lowering property, plant sterols such as β-sitosterol may also influence cholesterol-depending functions including its role in AD development. Using human platelets, a type of peripheral blood cells containing the most circulating amyloid precursor protein (APP), this study investigated the effect of β-sitosterol on high cholesterol-induced secretion of β amyloid protein (Aβ). It was found that β-sitosterol effectively inhibited high cholesterol-driven platelet Aβ release. In addition, β-sitosterol prevented high cholesterol-induced increase of activities of β- and γ-secretase, two APP cleaving enzymes to generate Aβ. Additional experiments showed that high cholesterol up-regulated lipid raft cholesterol. This effect of cholesterol could be suppressed by β-sitosterol. These findings suggest that β-sitosterol is able to inhibit high cholesterol-induced Aβ release probably through maintenance of membrane cholesterol homeostasis. Given that dietary plant sterols have the potential of penetrating the blood-brain barrier (BBB), these data suggest that plant sterols such as β-sitosterol may be useful in AD prevention.


Global dynamics of multiple delays within-host model for a hepatitis B virus infection of hepatocytes with immune response and drug therapy.
In this paper, a mathematical model describing the hepatitis B virus (HBV) infection of hepatocytes with the intracellular HBV-DNA containing capsids, cytotoxic T-lymphocyte (CTL), antibodies including drug therapy (blocking new infection and inhibiting viral production) with two-time delays is studied. It incorporates the delay in the productively infected hepatocytes and the delay in an antigenic stimulation generating CTL. We verify the positivity and boundedness of solutions and determine the basic reproduction number. The local and global stability of three equilibrium points (infection-free, immune-free, and immune-activated) are investigated. Finally, the numerical simulations are established to show the role of these therapies in reducing viral replication and HBV infection. Our results show that the treatment by blocking new infection gives more significant results than the treatment by inhibiting viral production for infected hepatocytes. Further, both delays affect the number of infections and duration i.e. the longer the delay, the more severe the HBV infection.


AICAr, a Widely Used AMPK Activator with Important AMPK-Independent Effects: A Systematic Review.
5-Aminoimidazole-4-carboxamide ribonucleoside (AICAr) has been one of the most commonly used pharmacological modulators of AMPK activity. The majority of early studies on the role of AMPK, both in the physiological regulation of metabolism and in cancer pathogenesis, were based solely on the use of AICAr as an AMPK-activator. Even with more complex models of AMPK downregulation and knockout being introduced, AICAr remained a regular starting point for many studies focusing on AMPK biology. However, there is an increasing number of studies showing that numerous AICAr effects, previously attributed to AMPK activation, are in fact AMPK-independent. This review aims to give an overview of the present knowledge on AMPK-dependent and AMPK-independent effects of AICAr on metabolism, hypoxia, exercise, nucleotide synthesis, and cancer, calling for caution in the interpretation of AICAr-based studies in the context of understanding AMPK signaling pathway.


The food environment in Latin America: a systematic review with a focus on environments relevant to obesity and related chronic diseases.
Food environments may be contributing to the rapid increase in obesity occurring in most Latin American (LA) countries. The present study reviews literature from LA that (i) describes the food environment and policies targeting the food environment (FEP); and (ii) analytic studies that investigate associations between the FEP and dietary behaviours, overweight/obesity and obesity related chronic diseases. We focus on six dimensions of the FEP: food retail, provision, labelling, marketing, price and composition. Systematic literature review. Three databases (Web of Science, SciELO, LILACS) were searched, from 1 January 1999 up to July 2017. Two authors independently selected the studies. A narrative synthesis was used to summarize, integrate and interpret findings. Studies conducted in LA countries. The search yielded 2695 articles of which eighty-four met inclusion criteria. Most studies were descriptive and came from Brazil (61 %), followed by Mexico (18 %) and Guatemala (6 %). Studies were focused primarily on retail/provision (n 27), marketing (n 16) and labelling (n 15). Consistent associations between availability of fruit and vegetable markets and higher consumption of fruits and vegetables were found in cross-sectional studies. Health claims in food packaging were prevalent and mostly misleading. There was widespread use of marketing strategies for unhealthy foods aimed at children. Food prices were lower for processed relative to fresh foods. Some studies documented high sodium in industrially processed foods. Gaps in knowledge remain regarding policy evaluations, longitudinal food retail studies, impacts of food price on diet and effects of digital marketing on diet/health.


[ABDOMINAL MASS AND ASCITES AS RISK FACTORS OF PELVIC ORGAN PROLAPSE].
(Objective) A rise of intra-abdominal pressure may exacerbate pelvic organ prolapse (POP) as well as abdominal hernias. This paper aims to assess the possible risk factors of an abdominal mass and ascites as comorbidities of POP. (Methods) We retrospectively reviewed the medical charts of 2,748 POP patients between 2010 and 2016 and extracted eight cases (0.3%) with abdominal mass and ascites as risk factors. (Results) All eight patients were multiparous women aged between 52 and 88 years old. Three patients (cases 1-3) were referred to us for surgery related to POP from gynecologists with previously undetected ovarian tumors. In case 1, we noticed abdominal distension during a transvaginal mesh (TVM) operation. Postoperative CT and MRI scans confirmed the presence of an ovarian tumor 24 cm in diameter (mucinous cystic tumor, borderline malignant). In case 2, transvaginal ultrasound could not detect the ovaries, but a transabdominal ultrasound, which was done to investigate urinary retention, revealed an ovarian tumor 18 cm in diameter (mucinous cystic adenoma). In case 3, a detailed patient history outlined the patient's sense of abdominal fullness and a transvaginal ultrasound found ovarian cancer 10 cm in diameter with ascites (serous adenocarcinoma). Case 4 suffered from autosomal dominant polycystic kidney disease (ADPKD) with large liver cysts. The patient underwent a TVM operation to treat the presenting POP with unusual bleeding (460 g). Case 5 had abdominal distension and cystocele due to huge abdominal mass (recurrence of malignant lymphoma); she desired conservative follow-up to tumor and POP due to old age (88 years old). Two patients suffered from end-stage cancer (case 6: colorectal cancer, case 7: breast cancer) with liver metastasis. In cases 6 and 7, the patients' POP worsened with the increase of ascites, which was managed conservatively. Case 8 presented with liver cirrhosis related ascites and a total uterine prolapse, simultaneously. Colpocleisis was cancelled due to the onset of hepatic coma. (Conclusions) Abdominal mass and ascites are risk factors of POP by increasing abdominal pressure and lesions such as ovarian tumors may present as POP. Even when POP patients are referred from gynecologists, a vaginal examination, carefully recorded patient history, and abdominal palpation should be included as part of a standard treatment regimen to reliably exclude underlying diseases.


Recent trends in aboriginal mortality.
Mortality is an important measure of the extent of the health disadvantages experienced by Australia's Aborigines. In the absence of routinely available data on Aboriginal deaths, this paper has collated information from a number of sources. In addition to published reports and theses, it also includes unpublished data provided to the Australian Institute of Health by State and Territory health authorities. The review also draws on data on maternal deaths, collated on a triennial basis and published by the National Health and Medical Research Council. Despite improvements in some indices of mortality, the death rates of Aborigines, particularly for young and middle-aged adults, are unacceptably higher than those of non-Aboriginal Australians. Without substantial reductions in death rates, the expectation of life of Aborigines will remain comparable with that of people living in developing countries.


Intrapartum intravenous fluids for caesarean delivery and newborn weight loss: a retrospective cohort study.
To examine weight loss (WL) and excess weight loss (EWL) among newborns of caesarean delivery, comparing colloids plus crystalloids versus crystalloids only. Also, to examine different doses of intrapartum intravenous fluids on WL and EWL. Comparative safety retrospective cohort study. University Teaching Hospital, Moncton, Canada. Mothers exposed to intravenous fluids with caesarean delivery between 2008 and 2016. Exposure to colloids plus crystalloids was compared with crystalloids only, and dose-response analyses were performed for colloids, crystalloids and total intravenous fluids doses. Linear and logistic regression models were used, adjusting for potential confounders. Infants' WL was measured at days 1, 2 and 3 post partum, and EWL defined as loss of >7% of birth weight. From 801 mother-infant pairs, 176 were exposed to colloids plus crystalloids and 625 were exposed to crystalloids only (overall mean birth weight=3416 g, EWL=2%, 41.4% and 55.5% on days 1, 2 and 3, respectively). No significant difference in newborns' WL was observed on any of the days assessed. Adjusted OR (95% CI) of EWL was 1.0 (0.3 to 3.3) at 24 hours, 1.0 (0.7 to 1.5) at 48 hours and 1.4 (0.9 to 2.2) at 72 hours. No dose-response relationship was detected with type-specific and total intravenous fluids exposures. The risk of EWL was similar with colloids plus crystalloids and crystalloids only, suggesting that both therapeutic options can be considered during caesarean delivery. The absence of dose-response relationships adds confirmatory evidence to the intravenous fluids safety profiles.


Downregulation of NLRP2 inhibits HUVEC viability by inhibiting the MAPK signaling pathway.
Nucleotide‑binding oligomerization domain (NOD)‑like receptor proteins (NLRPs) are a subfamily of NOD‑like receptors (NLRs) that mainly participate in innate immunity. Among the 14 NLRPs, studies on NLRP2 are few and mostly focus on its functions in reproduction and embryonic development. To the best of the authors' knowledge, there has been no research on the function of NLRP2 in human umbilical vein endothelial cells (HUVECs). The present study knockdown the expression of NLRP2 by transfecting a short interfering (si)RNA (siNLRP2) into HUVECs and investigating its effects on HUVECs. It was identified using a Cell Counting kit‑8 assay that knockdown of NLRP2 can inhibit cell proliferation in HUVECs. The results of wound healing and Transwell assays indicated that migration and invasion were also suppressed by siNLRP2 transfection in HUVECs. Flow cytometry demonstrated that siNLRP2 induced cell cycle arrest and apoptosis in HUVECs. Western blot analysis revealed that the expression levels of cell cycle and apoptosis‑associated proteins were markedly changed. In addition, knockdown of NLRP2 inhibited the mitogen‑activated protein kinase (MAPK) signaling pathway by elevating extracellular signal‑regulated kinase phosphorylation levels and reducing proto‑oncogene serine/threonine‑protein kinase expression. Taken together, it was concluded that NLRP2 served an important role in maintaining cell viability, proliferation and motility in HUVECs, mainly by promoting the MAPK signaling pathway.


Enhanced selective extraction of hexane from hexane/soybean oil mixture using binary gas mixtures of carbon dioxide.
Carbon dioxide (CO2) can effectively separate hexane from a mixture of soybean oil (SBO) and hexane with a slight coextraction of SBO. Previous research demonstrated that CO2 entrained with helium significantly reduced SBO solubility in CO2. In this study, CO2 was mixed with three gases (He, N2, or Ar) (0.5-30 vol %) to decrease SBO solubility while attempting to maintain hexane solubility. The binary gas mixtures (at 25 degrees C and 9.31 MPa) were passed through a 25 wt % hexane/SBO mixture inside a 2.5 m fractionation column. Coextracted SBO was inversely proportional to binary gas concentration, whereas residual hexane in the raffinate was proportional to binary gas concentration. The 10% binary mixture of N2 or Ar was the best compromise to obtain both low residual hexane levels (i.e., 26 ppm) and low SBO coextraction (i.e., only 40 mg). This carry-over of SBO represents a 95% reduction in SBO carry-over compared to neat CO2.


Investigating the role of beliefs about emotions, emotional suppression and distress within a pain management programme for fibromyalgia.
This study aims to explore the relationships between beliefs about emotions, emotional suppression, distress and global impact (i.e. the extent to which a patient's symptoms impact their life) in a longitudinal design with patients who are taking part in a pain management programme. A total of 40 participants with fibromyalgia took part in pain management programmes at multiple sites as part of their usual care in the National Health Service. Measures of beliefs about the unacceptability of experiencing and expressing emotions, emotional suppression, distress and global impact were completed before and after the programmes. Beliefs about emotions significantly reduced following treatment, but emotional suppression did not. Changes in beliefs about emotion correlated with changes in emotional suppression. Changes in distress were related to changes in suppression and the relationship between global impact and beliefs about emotions was approaching significance. Emotional suppression and beliefs about emotions may play a role in the improvement in distress following treatment. However, future research should examine these variables as mediators of the effect of treatment compared to waitlist controls in a larger sample.


Paraaortic sentinel lymph nodes: toward optimal detection and intraoperative localization using SPECT/CT and intraoperative real-time imaging.
Paraaortic sentinel node biopsy may be a challenging procedure because the sentinel nodes are located retroperitoneally in close proximity to vital structures. The purpose of this study was to describe and evaluate the value of preoperative SPECT/CT for lymphatic mapping, and a portable gamma-camera for intraoperative radioguidance, in patients with paraaortic sentinel nodes. We evaluated our practice in 18 patients, who were treated at The Netherlands Cancer Institute with sentinel lymphadenectomy for different urologic malignancies and showed paraaortic drainage on preoperative images. After intratumoral injection of (99m)Tc-nanocolloid, the patients underwent sequential planar lymphoscintigraphy, hybrid SPECT/CT, and sentinel lymphadenectomy. Intraoperative node detection and localization were guided by a laparoscopic gamma-probe and a portable gamma-camera. This gamma-camera was set to display both the (99m)Tc signal and the (125)I-seed signal. This (125)I seed was placed on top of the gamma-probe, functioning as a pointer on screen, thus enabling real-time sentinel node localization with the gamma-camera. In 16 patients with midabdominal drainage on planar images and in 2 patients with nonvisualization on planar images, SPECT/CT showed clear localization of paraaortic sentinel nodes in relation to the abdominal vessels. Five patients underwent open surgery, and 13 patients underwent laparoscopy. The paraaortic sentinel nodes were successfully localized and removed in 15 patients with the help of the portable gamma-camera and gamma-probe and in 3 patients with the gamma-probe only. In 1 patient, the paraaortic sentinel node showed a metastasis. If retroperitoneal drainage is expected, SPECT/CT provides good detection and clear localization of sentinel nodes in relation to anatomic structures. Detection and removal of paraaortic sentinel nodes by means of a laparoscopic gamma-probe and real-time imaging with a portable gamma-camera is a successful method with high intraoperative detection rates.


Association of a FAM13A variant with interstitial lung disease in Japanese rheumatoid arthritis.
Interstitial lung disease (ILD) occasionally occurs in rheumatoid arthritis (RA) and confers a dismal prognosis. We previously reported that a single-nucleotide variant (SNV) of MUC5B was associated with ILD in RA. However, the pathogenesis of ILD in Japanese patients with RA could not be explained solely by this SNV because its frequency is extremely low in the Japanese population. Here, we examined whether a different idiopathic pulmonary fibrosis susceptibility SNV might be associated with ILD in Japanese patients with RA. Genotyping of rs2609255 (G/T) in FAM13A was conducted in 208 patients with RA with ILD and 420 without chronic lung disease using TaqMan assays. A significant association with usual interstitial pneumonia (UIP) in RA was detected for rs2609255 under the allele model (p=0.0092, Pc=0.0276, OR 1.53, 95% CI 1.12 to 2.11) and recessive model for the G allele (p=0.0003, Pc=0.0009, OR 2.63, 95% CI 1.59 to 4.32). FAM13A rs2609255 was significantly associated with UIP in male patients with RA (p=0.0043, OR 3.65, 95% CI 1.52 to 8.73) under the recessive model. This study is the first to document an association of rs2609255 with ILD in Japanese patients with RA, implicating it in the pathogenesis of UIP, though studies on the function of rs2609255 are warranted.


Direct biosensor immunoassays for the detection of nonmilk proteins in milk powder.
The low prices of some nonmilk proteins make them attractive as potential adulterants in dairy products. An optical biosensor (BIACORE 3000) was used to develop a direct and combined biosensor immunoassay (BIA) for the simultaneous detection of soy, pea, and soluble wheat proteins in milk powders. Affinity-purified polyclonal antibodies raised against the three protein sources were immobilized in different flow channels (Fcs) on the biosensor chip (CM5). Dissolved milk powders were injected (20 microL injections at 20 microL min(-1)) through the serially connected Fcs, and the antibody-bound plant proteins were detected directly. The total run time between samples, including a regeneration step with 5 microL of 10 mM HCl, was 5 min. The limits of detection in milk powder were below 0.1% of plant protein in the total milk protein content. The antibodies also recognized some proteins from other plant sources, which made this BIA even more suitable as a broad screening assay for nonmilk proteins.


[Abdominal approaches and drainages of the abdominal cavity].
Appropriate access to the abdominal cavity is the first and crucial step for successful abdominal surgical intervention. In planning the incision, several variables have to be considered, such as anatomy of the abdominal wall, localization of the target organ, and individual conditions (previous incisions, minimal access surgery, etc). Medial laparotomy is the preferred incision for emergency cases and ill-defined pathologies, allowing access and hence exploration to all quadrants. Transverse laparotomies give superior access to the dorsal and right aspects of the liver and cause less pain in patients unfit for regional anesthetic procedures. Draining of the abdominal cavity is used after various resective and reconstructive procedures, but there is little evidence for its use in a number of operations such as gastric, hepatic, and colorectal resections. Advantages and disadvantages of different abdominal wall incisions and drainages are discussed.


Bioprospection of the trichomonacidal activity of lipid extracts derived from marine macroalgae Gigartina skottsbergii.
Algal extracts are sources of bioactive substances with applications in the development of novel alternative drugs against several diseases, including trichomoniasis sexually transmitted infection caused by Trichomonas vaginalis. Factors such as clinical failures and resistant strains limit the success of the existing drugs available for treating this disease. Therefore, searching for viable alternatives to these drugs is essential for the treatment of this disease. The present study was conducted for, in vitro and in silico characterization of extracts obtained from marine macroalgae Gigartina skottsbergii at stages gametophidic, cystocarpic, and tetrasporophidic. In addition, antiparasitic activity of these extracts against the ATCC 30236 isolate of T. vaginalis, their cytotoxicity, and gene expression of trophozoites after treatment were evaluated. The minimum inhibitory concentration and 50% inhibition concentration were determined for each extract. Results: In vitro analysis of the extracts' anti-T. vaginalis activity revealed an inhibitory effect of 100%, 89.61%, and 86.95% for Gigartina skottsbergii at stages gametophidic, cystocarpic, and tetrasporophidic, respectively, at 100 μg/mL. In silico analysis revealed the interactions between constituents of the extracts and enzymes from T. vaginalis, with significant free energy values obtained for the binding. None of the extract concentrations exhibited cytotoxic effects on VERO cell line compared to control, while cytotoxicity on HMVII vaginal epithelial cells line was observed at 100 μg/mL (30% inhibition). Gene expression analysis revealed differences in the expression profile of T. vaginalis enzymes between the extract-treated and control groups. According to these results, Gigartina skottsbergii extracts exhibited satisfactory antiparasitic activity.


Nontuberculous mycobacterial infections in heart transplant recipients: a seventeen-year experience.
A retrospective review was undertaken to determine the clinical features, outcome, and impact on survival of nontuberculous mycobacterial infections in 14 (of 502) heart transplant recipients. The prevalence of these infections was significantly higher (p less than 0.05) during the precyclosporine than during the cyclosporine era. The mean interval from transplantation to diagnosis was 3.5 +/- 0.7 years (+/- SEM). The 14 patients had a higher (p less than 0.05) linearized rejection rate than did other transplant patients during postoperative years 1, 2, and 4 to 6, and had received 7.3 +/- 2 gm of methylprednisolone as bolus treatment of rejection before diagnosis. Eight patients were initially seen with disseminated disease, four had localized pulmonary infection, one had subcutaneous infection in the previous site of a left ventricular assist device, and in one the organism was cultured from a fluid collection adjacent to a hip prosthesis. Twelve patients were first given 3 +/- 1 antimycobacterial drugs; the infections were usually controlled, but in 75% of patients the therapeutic regimen was prematurely stopped or altered because of drug toxicity. Mycobacterial infection was a contributory cause of death in only one patient. Actuarial survival in the 14 patients was not significantly different from the entire transplant population at 4 to 7 years. We conclude that nontuberculous mycobacterial infections occur late after heart transplantation, that drug treatment is usually successful (although difficult), and that long-term survival is not adversely affected if the infection is successfully controlled.


[A system, developed by prescribing doctors, of indicators of the quality of pharmaceutical prescription in primary care].
To work out a system of indicators and standards, designed by means of a consensus group of general practitioners, that enables evaluation of the quality of Primary Care prescription to be improved. Design. Informal consensus method. A PC area in Asturias with 156 614 inhabitants and 9 health centres.Participants. Nine PC doctors accepted voluntarily to take part in the project. A consensus group of nine PC doctors was formed. At a first meeting they worked out some potential indicators of quality of prescription. After applying these indicators to the prescription data, a second meeting was held, at which the results of applying the indicators along with criteria of excellence were used to establish the definitive indicators and their numerical values of optimum compliance. It was possible to agree by consensus 11 indicators of quality of prescription and their values of optimum compliance. Eight of the indicators referred to choice of a medicine within a pharmaco-therapeutic group; and three, to groups in which the volume of prescription may suggest improper usage. It is feasible for PC doctors to reach a consensus on a group of criteria that is perceived as valid for measuring quality of prescription and which includes certain impalpable values for determining quality.


Altered Regulation of HIF-1α in Naive- and Drug-Resistant EGFR-Mutant NSCLC: Implications for a Vascular Endothelial Growth Factor-Dependent Phenotype.
The treatment of patients with EGFR-mutant NSCLC with vascular endothelial growth factor (VEGF) inhibitors in combination with EGFR inhibitors provides a greater benefit than EGFR inhibition alone, suggesting that EGFR mutation status may define a patient subgroup with greater benefit from VEGF blockade. The mechanisms driving this potentially enhanced VEGF dependence are unknown. We analyzed the effect of EGFR inhibition on VEGF and HIF-1α in NSCLC models in vitro and in vivo. We determined the efficacy of VEGF inhibition in xenografts and analyzed the impact of acquired EGFR inhibitor resistance on VEGF and HIF-1α. NSCLC cells with EGFR-activating mutations exhibited altered regulation of VEGF compared with EGFR wild-type cells. In EGFR-mutant cells, EGFR, not hypoxia, was the dominant regulator of HIF-1α and VEGF. NSCLC tumor models bearing classical or exon 20 EGFR mutations were more sensitive to VEGF inhibition than EGFR wild-type tumors, and a combination of VEGF and EGFR inhibition delayed tumor progression. In models of acquired EGFR inhibitor resistance, whereas VEGF remained overexpressed, the hypoxia-independent expression of HIF-1α was delinked from EGFR signaling, and EGFR inhibition no longer diminished HIF-1α or VEGF expression. In EGFR-mutant NSCLC, EGFR signaling is the dominant regulator of HIF-1α and VEGF in a hypoxia-independent manner, hijacking an important cellular response regulating tumor aggressiveness. Cells with acquired EGFR inhibitor resistance retained elevated expression of HIF-1α and VEGF, and the pathways were no longer EGFR-regulated. This supports VEGF targeting in EGFR-mutant tumors in the EGFR inhibitor-naive and refractory settings.


Molecular Characteristics of Carnivore protoparvovirus 1 with High Sequence Similarity between Wild and Domestic Carnivores in Taiwan.
Carnivore protoparvovirus 1 (CPPV-1) is a DNA virus causing gastrointestinal disease and immunosuppression in various terrestrial carnivores. Domestic dogs and cats are considered the primary CPPV-1 reservoirs. The habitat overlap of wild carnivores and free-roaming dogs increases the threat of CPPV-1 transmission between them. This study explored the CPPV-1 distribution among wild carnivores in Taiwan through PCR screening and compared the partial capsid protein (VP2) gene sequences from wild and domestic carnivores. In total, 181 samples were collected from 32 masked palm civets (Paguma larvata), 63 Chinese ferret badgers (Melogale moschata), and 86 crab-eating mongooses (Herpestes urva), from 2015 to 2019 were screened for CPPV-1. The average prevalence of CPPV-1 was 17.7% (32/181), with the highest prevalence in masked palm civets (37.5%). In addition, a masked palm civet was coinfected with two CPPV-1 strains. Among the 33 partial VP2 gene sequences, 23 were identical to the sequences amplified from domestic dogs and cats in Asia, and the remaining 10 were identified for the first time. This study supported the circulation of CPPV-1 strains with the same genomic features as domestic carnivores that are also in wild carnivores from the same environment in Taiwan by molecular data. Therefore, further population control and health management of free-roaming domestic carnivores are recommended.


A single nucleotide polymorphism of macrophage migration inhibitory factor is related to inflammatory response in coronary bypass surgery using cardiopulmonary bypass.
Cardiac surgery causes induction and release of inflammatory mediators that may be regulated by genetic background. Macrophage migration inhibitory factor (MIF) is a proinflammatory mediator that is known to be up-regulated in patients undergoing cardiac operations. Here we analyzed genotype distribution and allele frequency of the MIF-173*G/C single nucleotide polymorphism (SNP) and MIF plasma levels in patients undergoing surgical revascularization with (on-pump, n=45) and without (off-pump, n=34) cardiopulmonary bypass (CPB). Genotyping was performed using a real-time PCR-based system with a hybridization probe system specific for the MIF-173*G/C SNP. In on-pump patients, blood samples were drawn before start of CPB, after termination of CPB and 12h postoperatively. In off-pump patients, blood samples were collected before stabilizer placement, after removal of the stabilizer and 12h postoperatively. MIF levels were measured using ELISA technique. Genotype distribution and allele frequencies were comparable between on-pump and off-pump patients. When comparing patients according to MIF genotype, a significant increase of MIF plasma levels after completed coronary bypass grafting using CPB was found in patients heterozygous for the MIF-173*G/C SNP (p<0.05). Moreover, on-pump patients showed significantly decreased MIF plasma levels after 12h postoperatively (p<0.05). In off-pump patients, MIF plasma levels were not significantly different over the time-course and were independent of the genotype. Patients carrying the C-allele showed significantly increased levels of the proinflammatory cytokine MIF compared to G/G homozygous when revascularization was carried out using CPB. The G/C genotype may be associated with a severe inflammatory reaction and therefore preoperative screening could be beneficial for patients undergoing cardiac surgery using CPB.


Identification of Signatures of Selection for Litter Size and Pubertal Initiation in Two Sheep Populations.
Fecundity is an important economic trait in sheep that directly affects their economic and productive efficiency. Our study aimed to identify SNP loci associated with sheep puberty or litter size which could be used in future breeding programs to improve fertility. Genomic DNA was obtained from Hetian and Cele Black sheep breeds and used for reduced-representation genome sequencing to identify SNP loci associated with pubertal initiation and litter size. Selective signatures analysis was performed based on the fixation index and nucleotide diversity, followed by pathway analysis of the genes contained in the selected regions. The selected SNP loci in the genes associated with pubertal initiation and litter size were validated using both sheep breeds. In total, 384,718 high quality SNPs were obtained and 376 genes were selected. Functional annotation of genes and enrichment analysis identified 12 genes associated with pubertal initiation and 11 genes associated with litter size. SNP locus validation showed that two SNP on PAK1 and four on ADCY1 may be associated with pubertal initiation, and one SNP on GNAQ gene (NC_040253.1: g.62677376G > A) was associated with litter size in Cele Black sheep. Our results provide new theoretical support for sheep breeding.


Retreatability of two endodontic sealers, EndoSequence BC Sealer and AH Plus: a micro-computed tomographic comparison.
Recently, bioceramic sealers like EndoSequence BC Sealer (BC Sealer) have been introduced and are being used in endodontic practice. However, this sealer has limited research related to its retreatability. Hence, the aim of this study was to evaluate the retreatability of two sealers, BC Sealer as compared with AH Plus using micro-computed tomographic (micro-CT) analysis. Fifty-six extracted human maxillary incisors were instrumented and randomly divided into 4 groups of 14 teeth: 1A, gutta-percha, AH Plus retreated with chloroform; 1B, gutta-percha, AH Plus retreated without chloroform; 2A, gutta-percha, EndoSequence BC Sealer retreated with chloroform; 2B, gutta-percha, EndoSequence BC Sealer retreated without chloroform. Micro-CT scans were taken before and after obturation and retreatment and analyzed for the volume of residual material. The specimens were longitudinally sectioned and digitized images were taken with the dental operating microscope. Data was analyzed using an ANOVA and a post-hoc Tukey test. Fisher exact tests were performed to analyze the ability to regain patency. There was significantly less residual root canal filling material in the AH Plus groups retreated with chloroform as compared to the others. The BC Sealer samples retreated with chloroform had better results than those retreated without chloroform. Furthermore, patency could be re-established in only 14% of teeth in the BC Sealer without chloroform group. The results of this study demonstrate that the BC Sealer group had significantly more residual filling material than the AH Plus group regardless of whether or not both sealers were retreated with chloroform.


Acute silent cerebral infarction in children with sickle cell anemia.
Silent cerebral infarctions (SCI) occur in up to 35% of children with sickle cell anemia (HbSS) but are rarely recognized during the initial 10-14 days when diffusion weighted magnetic resonance imaging (MRI) can differentiate acute infarctions from remote events. We report acute SCI in seven children with HbSS who had areas of restricted diffusion on MRI without persistent focal neurologic deficits. Four had acute SCI identified following acute anemic events. Our observations suggest that SCI are detectible in the acute phase, present with subtle neurologic symptoms, result in permanent neurologic injury, and may be caused by acute anemic events.


Conformationally Induced Off-On Two-Photon Fluorescent Bioprobes for Dynamically Tracking the Interactions among Multiple Organelles.
Unveiling the synergism among multiple organelles for fully exploring the mysteries of the cell has drawn more and more attention. Herein, we developed two two-photon fluorescent bioprobes (Lyso-TA and Mito-QA), of which the conformational change triggered an "off-on" fluorescent response. Lyso-TA can real-time monitor the fusion and movement of lysosomes as well as unveil the mitophagy process with the engagement of lysosomes. Mito-QA was transformed from Lyso-TA by one-step ambient temperature reaction, visualizing the dysfunctional mitochondria through a shift from mitochondria to nucleoli. With superior two-photon absorption cross section, good biocompatibility, and greater penetration depth, two small bioprobes were both applied in in vivo bioimaging of brain tissues and zebrafish.


Inducible aluminum resistance of Acidiphilium cryptum and aluminum tolerance of other acidophilic bacteria.
Aluminum ions are highly soluble in acidic environments. Toxicity of aluminum ions for heterotrophic, facultatively and obligately chemolithoautotrophic acidophilic bacteria was examined. Acidiphilium cryptum grew in glucose-mineral medium, pH 3, containing 300 mM aluminum sulfate [Al(2)(SO(4))(3)] after a lag phase of about 120 h with a doubling time of 7.6 h, as compared to 5.2 h of growth without aluminum. Precultivation with 1 mM Al(2)(SO(4))(3) and transfer to a medium with 300 mM Al(2)(SO(4))(3) reduced the lag phase from 120 to 60 h, and immediate growth was observed when A. cryptum was precultivated with 50 mM Al(2)(SO(4))(3), suggesting an aluminum-induced resistance. Aluminum resistance was not induced by Fe(3+) ions and divalent cations. Upon exposure of A. cryptum to 300 mM Al(2)(SO(4))(3), the protein profile changed significantly as determined by SDS-PAGE. When other acidophiles were cultivated with 50-200 mM aluminum sulfate, no lag phase was observed while the growth rates and the cellular yields were significantly reduced. This growth response was observed with Acidobacterium capsulatum, Acidiphilium acidophilum, Acidithiobacillus ferrooxidans, and Acidithiobacillus thiooxidans. Precultivation of these strains with aluminum ions did not alter the growth response caused by aluminum. The content of A. cryptum cultivated with 300 mM Al(2)(SO(4))(3)was 0.44 microg Al/mg cell dry weight, while that of the other strains cultivated with 50 mM Al(2)(SO(4))(3) ranged from 0.30 to 3.47 microg Al/mg cell dry weight.


Diameter selectivity of protein encapsulation in carbon nanotubes.
Biomolecular-carbon nanotube (CNT) complexes are of great importance in biological and biomedical devices, and recently spontaneous encapsulation of biomolecules into CNTs has attracted great interest. In this work, we explored the diameter selectivity of the protein encapsulation in CNTs via molecular dynamics simulations, and the free energy changes of the systems were calculated for mechanism exploration. It is proved that there is an optimal tube size which provides the most effective encapsulation for a given protein molecule, and the encapsulations in the overlarge and overcrowded tubes are hindered by different factors based on the analysis of system energy contribution. In addition, the significance of the solvents for the system is also of concern.


Next Step in Gene Delivery: Modern Approaches and Further Perspectives of AAV Tropism Modification.
Today, adeno-associated virus (AAV) is an extremely popular choice for gene therapy delivery. The safety profile and simplicity of the genome organization are the decisive advantages which allow us to claim that AAV is currently among the most promising vectors. Several drugs based on AAV have been approved in the USA and Europe, but AAV serotypes' unspecific tissue tropism is still a serious limitation. In recent decades, several techniques have been developed to overcome this barrier, such as the rational design, directed evolution and chemical conjugation of targeting molecules with a capsid. Today, all of the abovementioned approaches confer the possibility to produce AAV capsids with tailored tropism, but recent data indicate that a better understanding of AAV biology and the growth of structural data may theoretically constitute a rational approach to most effectively produce highly selective and targeted AAV capsids. However, while we are still far from this goal, other approaches are still in play, despite their drawbacks and limitations.


Relative roles of upper and lower lacrimal canaliculi in normal tear drainage.
A total of 88 cases of impaction of the eyelash into the lacrimal punctum were collected from 34,256 outpatients, and 39 cases of lash impaction into the Meibomian gland duct were found among 30,456 outpatients. Observations of these patients were analyzed to find the relative roles of the upper and lower lacrimal canaliculi in the normal tear drainage. In the cases of the Meibomian gland duct, the impaction occurred at a similar incidence in the upper and lower eyelids. However, in the cases of the punctum, the upper punctum was affected about three times more frequently than the lower punctum. The impaction of the eyelashes probably occurred at about the same rate in both puncta, but the present results suggested that the lashes in the lower punctum disappeared more easily into the drainage system than the lashes in the upper punctum. The lower canaliculus should, therefore, play the major role in normal tear drainage.


Surgical Treatment of Osteochondral Lesions of the Tibial Plafond: A Systematic Review and Meta-Analysis.
The literature on osteochondral lesions of the tibial plafond (OLTPs) is sparse. The aim of this study was therefore to provide an overview of clinical and radiological outcomes following treatment of OLTPs. We performed a systematic search of the MEDLINE, Embase, and Cochrane library databases. The review was performed in accordance with the PRISMA (Preferred Reporting Items for Systematic reviews and Meta-Analyses) guidelines and included all original articles on treatment outcomes for OLTPs. The methodological quality of the articles was assessed using the Methodological Index for Non-Randomized Studies (MINORS). Baseline patient and lesion characteristics were pooled and weighted according to the number of lesions per study. The primary outcome was any clinical or patient-reported outcome measure pooled by treatment method when separable data were available. Secondary outcomes were complications, reoperation rates, radiological outcomes, and sport outcomes. The search yielded 2,079 articles, of which 10 studies (1 prospective case series, 1 retrospective comparative study, and 8 retrospective case series) with a total of 175 patients were included. The overall methodological quality of the studies was low. All patients were treated surgically; 96% of the lesions were primary cases (i.e., first-time surgery) and 58% were solitary tibial lesions (i.e., no opposing talar lesion). Arthroscopic bone marrow stimulation was the most frequently used treatment strategy (51%), followed by cartilage transplantation (17%), chondrogenesis-inducing techniques (11%), osteochondral transplantation (3%), retrograde drilling (3%), and mixed (i.e., inseparable) treatments (15%). The clinical outcomes of the different surgical therapies were considered to be moderate to good. The pooled postoperative AOFAS (American Orthopaedic Foot & Ankle Society) score for bone marrow stimulation and osteochondral transplantation was 54.8 (95% confidence interval [CI], 49.5 to 85.0) (n = 14) and 85.3 (95% CI, 56 to 100) (n = 3), respectively. Overall, complications and reoperations were rarely reported. The pooled complication and reoperation rates could only be calculated for bone marrow stimulation and were 5% and 7%, respectively. Surgical interventions for OLTPs appear to yield moderate to good clinical outcomes. Bone marrow stimulation resulted in a moderate AOFAS score. Complications and reintervention rates were found to be low. The current evidence in the literature is limited because of the underreporting of clinical, radiological, and sport data and the heterogenous outcome scores reported. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Successful extraction of refractory thrombus from an ectatic coronary artery using stent retriever during primary angioplasty for acute myocardial infarction: a case report.
Ectatic coronary segments are nidi for thrombus formation due to altered flow dynamics and stasis-an important component of Virchow's triad. Ectasia accompanied by an adjacent coronary stenosis with or without a plaque event can lead to acute myocardial infarctions complicated by huge thrombus burden. Here, we present a case of a young male with acute inferior wall myocardial infarction complicated by Type IV coronary artery ectasia of the right coronary artery and huge thrombus burden that was refractory to conventional methods of thrombus management like thrombo-suction and intracoronary cocktails including tenecteplase. After a stormy course of transient complete heart block and recurrent ventricular tachycardia, the thrombus was successfully extracted using a stent retriever to achieve thrombolysis in myocardial infarction (TIMI) II plus flow distally. The lesion just distal to the ectasia was stented 24 h later to achieve TIMI III flow and the patient had an uneventful recovery subsequently. Angiographically visible thrombus that is refractory to intracoronary medications and aspiration thrombectomy can be successfully managed using a stent retriever.


Associations between dietary patterns and demographics, lifestyle, anthropometry and blood pressure in Chinese community-dwelling older men and women.
This cross-sectional study examined dietary patterns, and the associations of these patterns with demographics, lifestyle, anthropometry and blood pressure in 3707 Chinese people aged 65 years and above taking part in a population-based cohort study investigating the risk factors for osteoporosis. Baseline dietary data were collected using a validated FFQ. Dietary patterns were identified using factor analysis. Scores were calculated for each pattern. Demographics, lifestyle factors and self-reported hypertension history were collected through a questionnaire. BMI, waist circumference (WC), hip circumference (HC), waist-to-hip ratio, systolic blood pressure and diastolic blood pressure (DBP) were measured. Three dietary patterns were identified, namely 'vegetables-fruit', 'snacks-drinks-milk products' and 'meat-fish'. Participants who were more physically active, more educated, non-smokers and non-drinkers were more likely to have higher 'vegetables-fruit' dietary pattern scores. Current smoking habit and alcohol use were associated with higher 'snacks-drinks-milk products' dietary pattern scores and 'meat-fish' dietary pattern scores. 'Vegetables-fruit' dietary pattern scores were inversely (unstandardised regression coefficient B = -0·60 mmHg, 95 % CI -1·04, -0·16) and 'snacks-drinks-milk products' dietary pattern scores were positively (B = 0·50 mmHg, 95 % CI 0·08, 0·92) associated with DBP in men in multiple regressions. Higher 'meat-fish' dietary pattern scores were associated with higher BMI (B = 0·19 kg/m(2), 95 % CI 0·06, 0·33), waist-to-hip ratio (B = 0·004, 95 % CI 0·002, 0·007) and WC (B = 0·57 cm, 95 % CI 0·18, 0·97) in men, and higher BMI (B = 0·40 kg/m(2), 95 % CI 0·22, 0·57), WC (B = 0·87 cm, 95 % CI 0·39, 1·36) and HC (B = 0·61 cm, 95 % CI 0·26, 0·96) in women in multiple regressions. The influence of demographic and lifestyle characteristics on dietary patterns and the health risks associated with dietary patterns provides insights for the provision of tangible dietary advice to this population.


Application of camera phones in telehaematology.
We investigated the use of camera phones for telehaematology. First, the minimum requirements for the camera phones to be used in telehaematology were investigated. A single image containing white cells, red cells and platelets was sent from a camera phone to 33 different camera phones. Nine of the camera phones were found to be unsuitable for telehaematology due to low display resolution or no zoom function of the image. Then we examined the agreement between a haematologist using a suitable camera phone for remote diagnosis and the blood film report made in the usual way. Blood samples were collected from nine patients who had conditions in which diagnostically important morphological abnormalities occurred. In seven of the nine cases, the telehaematology responses were similar to the documented blood film reports. We conclude that telehaematology using camera phones offers a quick and potentially valuable method of support for the diagnostic haematology laboratory.


[Prevention of early respiratory complications in esophageal surgery by ventilation of the independent lungs].
Independent lung controlled ventilation by a double lumen tube has a beneficial effect in oesophagus surgery. Use of a tidal volume and an end-expiratory pressure different for each lung produces a drastic reduction of chest-X-ray abnormalities. In an homogen group of ten patients studied before and treated with conventional respiratory support, chest-X-ray abnormalities were seen in 80% cases. In this group of 34 patients treated with independent lung ventilation the rate of abnormalities is only of 20%. Independent lung ventilation decreases early pulmonary complications in dependent regions of the lungs, and late pulmonary complications in non dependent regions. This form of mechanical ventilation is performed with a "prototype" ventilator of small size, which permits synchronized or independent ventilation of the lungs.


Cationic intermediates in oxidative addition reactions of Cl2 to [PtCl2(cis-1,4-DACH)].
Oxidative addition and reductive elimination are fundamental processes in transition-metal chemistry. New interest in this field has been generated by the exploitation of platinum(IV) complexes as antitumor drugs. The two extra ligands can be used to render these species more resistant to attack by biological nucleophiles compared to their platinum(II) counterparts, to anchor additional pharmacologically active moieties, or, finally, to target the drug to specific sites by conferring responsiveness to some type of chemotaxis. On the other hand, platinum(IV) species are considered to be prodrugs and to require reduction to Pt(II) to become active. Thus, reductive elimination promoted by biological reducing agents becomes an important issue and it too could be exploited for targeting purposes. In this paper, we investigated the oxidation step in more detail and shown that, independent of the solvent used, a solvent molecule assists the reaction by entering in a trans position with respect to the attacking oxidant. In the case of bifunctional solvent molecules, such as dimethylsulfoxide, both S- and O-coordinated species are formed, the latter being thermodynamically favored. The substitution of the axially coordinated solvent molecule by a free chloride ion is found to be quite slow in organic solvents, as well as in water. It is also shown that the intermediate solvato-species can be exploited for binding just one molecule of another substrate in the axial position.


Risk of cholangiocarcinoma in patients with primary sclerosing cholangitis: diagnosis and surveillance.
Primary sclerosing cholangitis (PSC) is associated with an increased risk of hepatobiliary and extrahepatic malignancy. Particularly the risk of cholangiocarcinoma (CCA) is greatly increased. To provide potentially curative treatments for affected patients an early diagnosis of CCA is crucial. We here review the current advances with respect to CCA diagnosis and surveillance and discuss a rational approach on how to perform surveillance of CCA in PSC patients. Given the shortcomings of the current modalities for the surveillance and diagnosis of CCA in PSC, recent studies have focused on novel biomarkers for CCA. These include serum biomarkers (e.g., circulating angiopoeitin-2, cytokeratin-19 fragments, and antiglycoprotein 2 IgA autoantibodies, microRNA) as well as proteomics obtained from urine and/or bile. Novel approaches that may enhance the diagnostic value of brush cytology in future include the optimization of fluorescence in situ hybridization probes and the assessment of genetic aberrations. In addition, studies on advanced techniques (e.g., single-operator cholangioscopy and probe-based confocal laser endomicroscopy) have shown promising results with respect to CCA detection. Despite recent advances in the diagnosis of CCA in PSC, the detection of early-stage CCA remains difficult. A better understanding of CCA pathogenesis and large prospective studies on novel biomarkers and techniques are required to timely diagnose CCA in the future.


Spectral Reflectance Estimation Using Gaussian Processes and Combination Kernels.
This paper explores hyperspectral reflectance factor estimation using Gaussian process regression with multispectral- and trichromatic measurements. Estimations are performed in visible- (400-700 nm) or visible-near infrared (400-980 nm) wavelength ranges using the learning-based approach, where sensor and light spectral characteristics are not required. We first construct new estimation models via Gaussian processes, show connection to previous kernel-based models, and then evaluate new models by using marginal likelihood optimization within the probabilistic interpretation. By using standard spectral ensembles and several images in experiments, we evaluate new models with anisotropic radial- and combination kernels (process covariance), marginal likelihood optimization (parameter selection), as well as with input data transformations (pre-processing). Several new Gaussian process models provide spectral accuracy improvements for simulated and real data, when compared with the previous kernel-based models. Most versatile new model is using spectral subspace coordinate learning and combination kernels, and can be efficiently optimized via marginal likelihood. Preliminary results suggest that new models provide uncertainty estimates, which can be used for iterative training set augmentation.