The effect of block copolymer structure on the internalization of polymeric micelles by human breast cancer cells.
The objective of this study was to assess the effect of hydrophilic/hydrophobic block chain lengths on the internalization of poly(ethylene oxide)-block-poly(epsilon-caprolactone) (PEO-b-PCL) micelles by cancer cells. PEO-b-PCL block copolymers with varied PEO and PCL chain lengths were synthesized, assembled to polymeric micelles and loaded with a hydrophobic fluorescent probe (DiI) through a co-solvent evaporation method of physical encapsulation. The slow release of the fluorescent probe from the micellar structure was evidenced following DiI transfer to lipid vesicles. The extent of micellar uptake by cancer cells was investigated through their incubation with MCF-7 cells followed by measurement of the fluorescent emission intensity of DiI (lambda=550 nm) in separated lysed cells. Cellular internalization of polymeric micelles was confirmed by laser scanning microscopy. The mechanism of micellar uptake was investigated by pretreatment of MCF-7 cells with chlorpromazine and cytochalasin B. Encapsulation of DiI in PEO-b-PCL micelles lowered the extent and rate of hydrophobic probe internalization by cancer cells. For polymeric micelles with 5000 gmol(-1) of PCL and varied PEO molecular weights of 2000, 5000 and 13,000 gmol(-1), maximum uptake was observed at a PEO molecular weight of 5000 gmol(-1). For polymeric micelles with 5000 gmol(-1) of PEO and varied PCL molecular weights of 5000, 13,000 and 24,000 gmol(-1), maximum uptake was observed at 13,000 gmol(-1) of PCL. Chlorpromazine reduced the cellular uptake of PEO-b-PCL micelles independent from the block copolymer structure, pointing to the involvement of clathrin mediated endocytosis mechanisms in the uptake of polymeric micelles by cancer cells. Inhibition of cellular uptake of PEO-b-PCL micelles by cytochalasin B, on the other hand, was found to be dependent on the chemical structure of the core/shell forming blocks.


Transcriptome analysis of sRNA responses to four different antibiotics in Pseudomonas aeruginosa PAO1.
A large number of evidence showed that regulatory sRNAs could modulate antibiotic resistance and sensitivity. In this study, we used RNA-sequencing to profile sRNAs in wild type and antibiotic-resistant PAO1 selected by four antibiotics (polymyxin B, ciprofloxacin, doxycycline, and ceftriaxone). Totally, we found 113, 25, 91 and 12 differentially expressed sRNAs in polymyxin B-, ciprofloxacin-, doxycycline-, and ceftriaxone-resistant P. aeruginosa, respectively. To elucidate functions of differentially expressed sRNA, we predicated their target genes and obtained pathways enriched by their target genes. In addition, our results indicated that the downregulated sRNA spae884.1, spae3443.1, and spae5681.1 might involve in polymyxin B resistance by enhancing their target genes arnA, arnD, and arnT expression in PAO1, respectively. The upregulated sRNA spae3443.1 and spae649.2 might implicate in ciprofloxacin resistance by promoting their target gene pslK expression to increase biofilm formation in PAO1. The upregulated spae1558.1 might increase oprJ expression, as well as spae3959.1 and spae3706.1 might increase mexC expression to modulate doxycycline resistance in PAO1. The sRNA novel-N714 might involve in virulence in ceftriaxone-resistant PAO1 by activating its target gene PA1429 expression. Our study might provide bases of the underlying mechanism of sRNA in regulating antibiotic resistance of PAO1 against different antibiotics.


Analysis of IKBKG/NEMO gene in five Japanese cases of incontinentia pigmenti with retinopathy: fine genomic assay of a rare male case with mosaicism.
Incontinentia pigmenti (IP) is an X-linked dominant genodermatosis that is usually lethal in utero in males, though exceptionally they survive very rarely either with Klinefelter syndrome or a somatic mosaicism. We performed genomic analysis of five Japanese IP patients including a rare boy case, all of whom were definite cases with retinopathy. Four patients including the boy revealed the recurrent exon 4-10 deletion in the sole known causative gene IKBKG/NEMO, which was confirmed by various specific PCR techniques. The boy's saliva DNA showed a mosaicism consisting of the deletion and intact alleles, but his blood DNA did not. Relative quantification analysis of the real-time PCR data by ∆∆CT method estimated the mosaicism ratio of the boy's saliva as 45:55 (deletion:intact). A genomic analysis for the recurrent deletion at the nucleotide sequence level has been performed directly using patient's DNA and it has been clarified that the breakpoints are within two MER67B repeats in the intron 3 and downstream of exon 10. This is the first report of the assay for the mosaicism ratio of a male IP case with a recurrent exon 4-10 deletion of IKBKG/NEMO and the sequencing analysis of the breakpoints of the recurrent deletion directly using patient's sample.


X-ray photoelectron spectroscopic studies of hydrophilic surfaces modified via admicellar polymerization.
Admicellar polymerization with styrene monomer was used to coat the surface of two porous solids: titanium dioxide and alumina. X-ray photoelectron spectroscopy (XPS) measurements clearly indicate that after admicellar polymerization, organic material and surfactant are present on the surface of the solids. Water washes, performed immediately after admicellar polymerization, were successful in removing approximately 30% of the organic material, presumably mostly surfactant. The water wash was followed by Soxhlet extraction with toluene, with no measurable removal of organic material according to ignition loss measurements of the solid, gel permeation chromatography of the extracting solvent and also UV/VIS spectroscopy of the extracting solvent. However, there was a 5-15% drop in organic material after Soxhlet extraction according to XPS measurements. This difference is attributed to the difference in sampling; XPS samples only the exterior surface area while these bulk measurements sample both the interior and exterior surface area. This study details the ability of different washing steps to remove materials from the porous substrates used in this study.


Effects of phytoestrogen genistein on myocardial ischemia/reperfusion injury and apoptosis in rabbits.
To study the effect of genistein (GST) on rabbit heart ischemia/reperfusion (I/R) injury. Rabbit heart I/R injury was induced by occluding the left anterior descending coronary artery for 45 min and reperfusing for 180 min. GST (1.0 mg/kg) was intravenously injected 5 min before heart ischemia. Hemodynamic data, infarct size, and cardiomyocytic apoptosis were measured. The pathologic changes of I/R myocardium were observed. During the I/R, heart rate, mean arterial blood pressure, myocardial oxygen consumption, left ventricular (LV) -dp/dtmax and +dp/dtmax were decreased progressively. The infarct size was occupied 60.23 %+/-3.97 % (% of area at risk) in vehicle +I/R group while GST reduced the infarct size to 39.62 %+/-4.30 % (P<0.01). DNA ladder pattern in myocardium was revealed by agarose gel electrophoresis in vehicle +I/R group while was not found in GST+I/R group. Apoptotic cardiomyocytes were sparse within ischemic myocardium at risk in GST+I/R group as compared with that in vehicle +I/R group (TUNEL stain). Apoptosis rate in ischemic myocardium from vehicle +I/R and GST+I/R groups detected by flow cytometry were 15.33 %+/-1.31 % and 3.88 %+/-0.33 %, respectively. Fas and Bax protein expressions in ischemic myocardium of vehicle +I/R group were higher than that in GST+I/R group (P<0.01). Bcl-2/Bax ratio in vehicle +I/R group was lower than that in nonischemic myocardium (P<0.01), while in GST+I/R group, the Bcl-2/Bax ratio was higher than that in vehicle +I/R group (P<0.01). GST reduced infarct size and apoptosis of myocytes in I/R rabbit heart.


Comparison between the predictions of diffusion-reaction models and localized Ca2+ transients in amphibian skeletal muscle fibers.
We developed a three-dimensional cylindrical diffusion-reaction model of a single amphibian myofibril in which Ca(2+) release occurred only at the Z-line. The model incorporated diffusion of Ca(2+), Mg(2+), and all relevant buffer species, as well as the kinetic binding reactions between the buffers and appropriate ions. Model data was blurred according to a Gaussian approximation of the point spread function of the microscope and directly compared with experimental data obtained using the confocal spot methodology. The flux parameters were adjusted until the simulated Z-line transient matched the experimental one. This model could not simultaneously predict key parameters of the experimental M- and Z-line transients, even when model parameters were adjusted to unreasonably extreme values. Even though the model was accurate in predicting the Z-line transient under conditions of high [EGTA], it predicted a significantly narrower Ca(2+) domain than observed experimentally. We modified the model to incorporate a broader band of release centered at the Z-line. This extended release model was superior both in simultaneously predicting critical features of the Z- and M-line transients as well as the domain profile under conditions of high [EGTA]. We conclude that a model of release occurring exclusively at the Z-line cannot explain our experimental data and suggest that Ca(2+) may be released from a broader region of the sarcoplasmic reticulum than just the T-tubule-sarcoplasmic reticulum junction.


Equal channel angular extrusion of ultra-high molecular weight polyethylene.
Ultra-high molecular weight polyethylene (UHMWPE), a common bearing surface in total joint arthroplasty, is subject to material property tradeoffs associated with conventional processing techniques. For orthopaedic applications, radiation-induced cross-linking is used to enhance the wear resistance of the material, but cross-linking also restricts relative chain movement in the amorphous regions and hence decreases toughness. Equal Channel Angular Extrusion (ECAE) is proposed as a novel mechanism by which entanglements can be introduced to the polymer bulk during consolidation, with the aim of imparting the same tribological benefits of conventional processing without complete inhibition of chain motion. ECAE processing at temperatures near the crystalline melt for UHMWPE produces (1) increased entanglements compared to control materials; (2) increasing entanglements with increasing temperature; and (3) mechanical properties between values for untreated polyethylene and for cross-linked polyethylene. These results support additional research in ECAE-processed UHMWPE for joint arthroplasty applications.


Concurrent Cetuximab-based bioradiotherapy versus Cisplatin-based Chemoradiotherapy in the Definitive Management of Favourable Biology Human Papillomavirus-associated Oropharyngeal Squamous Cell Carcinoma: Systematic Review and Meta-analysis.
Replacing cisplatin with cetuximab concurrently during radiotherapy has been one of the strategies of treatment de-escalation in human papillomavirus (HPV)-associated oropharyngeal squamous cell carcinoma (OPSCC). However, until recently, there were limited data on the efficacy and safety of such an approach. A systematic search of the literature was carried out to identify prospective randomised controlled trials comparing definitive cisplatin-based chemoradiotherapy (CT-RT) versus cetuximab-based bioradiotherapy (BRT) in HPV-positive OPSCC. Overall survival and locoregional control were primary outcomes of interest; rates of acute and late toxicities (≥grade 3) were secondary end points. Outcome data were aggregated using a random-effects model as per Cochrane methodology including risk of bias assessment and expressed as hazard ratio or risk ratio as appropriate with respective 95% confidence intervals. Data from five randomised controlled trials involving 1560 patients with HPV-positive OPSCC were aggregated in the meta-analysis. Cetuximab-based BRT was associated with a significantly increased risk of death (hazard ratio = 2.83, 95% confidence interval 1.22-6.57; P = 0.02) and locoregional relapse (hazard ratio = 2.78, 95% confidence interval 1.77-4.39; P < 0.0001) compared with cisplatin-based CT-RT. Cisplatin was associated with higher rates of acute ≥grade 3 toxicity in terms of acute kidney injury, dry mouth, febrile neutropenia, hearing impairment, nausea and vomiting, whereas dermatitis and acneiform rash were more common with cetuximab. There were no significant differences in overall rates of late ≥grade 3 toxicity (risk ratio = 0.63, 95% confidence interval = 0.36-1.10; P = 0.10). In conclusion, there is moderate-certainty evidence that cetuximab-based BRT leads to inferior efficacy outcomes compared with cisplatin-based CT-RT in the definitive curative-intent management of HPV-associated OPSCC.


Mesenchymal Stem Cell in Mice Uterine and Its Therapeutic Effect on Osteoporosis.
Osteoporosis is a silent disease caused by low bone mineral density and is complicated by fractures. This study was designed to examine the differentiation of uterine stem cell-derived osteoprogenitor cells (UOPCs) both in vitro and in vivo, assessing their effectiveness in treating osteoporosis. CD271+/CD45- UOPCs were isolated from the endometrial tissue of inbred Balb/c mice through magnetic activated cell sorting. Stem cell differentiation assays were used for CD271+/CD45- UOPCs in vitro. In vivo, the UOPCs were implanted into mouse osteoporosis models through tail-vein injection for 8 weeks. Osteogenic differentiation was examined by X-rays and computed tomography (CT) scans. Enhanced green fluorescent protein (EGFP)-labeled UOPCs, obtained from C57BL/6-Tg (ACTb-EGFP) 1Osb/J mice, were used to assess cell survival in the osteoporosis model. The levels of osteogenic markers were assessed by enzyme-linked immunosorbent assay. In vitro, UOPCs were able to form into typical spheres and various differentiations. In vivo, implantation of UOPCs into osteoporosis model significantly increased bone mineral densities and bone microstructure parameters. The levels of a biochemical marker of bone metabolism, Semaphorin-3A, increased significantly. However, levels of receptor activator of nuclear factor kappa-B ligand decreased. Immunofluorescence staining of osteoporosis mice injected with green fluorescent protein+ UOPCs showed their survival for up to 7 days. In conclusion, stem cells with osteogenic differentiation potential can be isolated from uterine or endometrial tissue. These UOPCs can stably proliferate and differentiate in vitro or in vivo, which can inhibit bone resorption and osteoclast marker expression. In vivo, UOPCs significantly improved reduction in bone density caused by reduced estrogen levels. Such cell transplantation approach is potentially useful in the treatment of osteoporosis.


Evidence of discontinuity between psychosis-risk and non-clinical samples in the neuroanatomical correlates of social function.
Social dysfunction is a major feature of clinical-high-risk states for psychosis (CHR-P). Prior research has identified a neuroanatomical pattern associated with impaired social function outcome in CHR-P. The aim of the current study was to test whether social dysfunction in CHR-P is neurobiologically distinct or in a continuum with the lower end of the normal distribution of individual differences in social functioning. We used a machine learning classifier to test for the presence of a previously validated brain structural pattern associated with impaired social outcome in CHR-P (CHR-outcome-neurosignature) in the neuroimaging profiles of individuals from two non-clinical samples (total n = 1763) and examined its association with social function, psychopathology and cognition. Although the CHR-outcome-neurosignature could be detected in a subset of the non-clinical samples, it was not associated was adverse social outcomes or higher psychopathology levels. However, participants whose neuroanatomical profiles were highly aligned with the CHR-outcome-neurosignature manifested subtle disadvantage in fluid (PFDR = 0.004) and crystallized intelligence (PFDR = 0.01), cognitive flexibility (PFDR = 0.02), inhibitory control (PFDR = 0.01), working memory (PFDR = 0.0005), and processing speed (PFDR = 0.04). We provide evidence of divergence in brain structural underpinnings of social dysfunction derived from a psychosis-risk enriched population when applied to non-clinical samples. This approach appears promising in identifying brain mechanisms bound to psychosis through comparisons of patient populations to non-clinical samples with the same neuroanatomical profiles.


CD47-targeted cancer immunogene therapy: Secreted SIRPα-Fc fusion protein eradicates tumors by macrophage and NK cell activation.
CD47 protects healthy cells from macrophage attack by binding to signal regulatory protein α (SIRPα), while its upregulation in cancer prevents immune clearance. Systemic treatment with CD47 antibodies requires a weakened Fc-mediated effector function or lower CD47-binding affinity to prevent side effects. Our approach combines "the best of both worlds," i.e., maximized CD47 binding and full Fc-mediated immune activity, by exploiting gene therapy for paracrine release. We developed a plasmid vector encoding for the secreted fusion protein sCV1-hIgG1, comprising highly efficient CD47-blocking moiety CV1 and Fc domain of human immunoglobulin G1 (IgG1) with maximized immune activation. sCV1-hIgG1 exhibited a potent bystander effect, blocking CD47 on all cells via fusion protein secreted from only a fraction of cells or when transferring transfection supernatant to untransfected cells. The CpG-free plasmid ensured sustained secretion of sCV1-hIgG1. In orthotopic human triple-negative breast cancer in CB17-severe combined immunodeficiency (SCID) mice, ex vivo transfection significantly delayed tumor growth and eradicated one-third of tumors. In intratumoral transfection experiments, CD47 blockage and increased migration of macrophages into the tumor were observed within 17 h of a single injection. Natural killer (NK) cell-mediated lysis of sCV1-hIgG1-expressing cells was demonstrated in vitro. Taken together, this approach also opens the opportunity to block, in principle, any immune checkpoints.


[Multiple hemangiomas, polyposis coli, endometrial and papillary thyroid cancer].
A 27-year-old female patient presented with multiple lipomas and cavernous hemangiomas of the skin, hemangioma of the parotid gland, polyps of the gastrointestinal tract and endometrial carcinoma. After 1 year the patient presented with papillary thyroid carcinoma and a diagnosis of Cowden syndrome (CS) was made. Genetic testing revealed a pathogenic PTEN gene mutation. Hereditary tumor syndromes, such as CS are underdiagnosed. Clinical management can be adjusted to the needs of CS patients only if an early diagnosis is made. In CS the risk for thyroid, breast and endometrial cancer is severely increased.


Blood-nerve barrier studies in experimental allergic neuritis.
The integrity of the blood-nerve barrier (BNB) was studied during the development of experimental allergic neuritis (EAN). Lewis rats immunized with bovine nerve or myelin plus complete Freund's adjuvant developed histological lesions of EAN in nerve roots by 10-12 days and in sciatic nerves by 12-14 days. Evans blue-albumin (EBA) and horseradish peroxidase (HRP) were injected i.v. 1 h prior to killing on days 6-18. Perivascular and diffuse endoneurial leakage of the tracers was seen in nerve roots by 10-12 days post immunization (p.i.) and in sciatic nerves by 12-14 days. This coincided with the appearance of endoneurial infiltration with inflammatory cells and endoneurial proteinaceous edema at a time when Schwann cell and myelin changes were still minimal. Therefore, an alteration in BNB permeability occurs early in EAN, coincident with inflammatory cell infiltration. This could be an expression of delayed hypersensitivity, yet it would also facilitate the entry of anti-myelin antibodies into the endoneurium where they could initiate demyelination.


There is more: Intrasexual competitiveness, physical dominance, and intrasexual collaboration.
It is emphasized that in organizational settings, the responses to same-sex attractive others are enhanced among individuals high in intrasexual competitiveness; that especially attractive rivals who are perceived as unfriendly will induce competition; that among males, physical dominance may induce more competition than physical attractiveness; and that especially males may prefer to associate with attractive same-sex others for intrasexual collaboration.


HMGA2 is commonly expressed in uterine serous carcinomas and is a useful adjunct to diagnosis.
Serous carcinoma is the prototype of type 2 uterine carcinoma. In many cases, establishing a diagnosis is straightforward, but problems can arise in that papillary variants of endometrioid carcinoma may be mistaken for serous carcinoma, and glandular variants of serous carcinoma may be misdiagnosed as endometrioid carcinoma. Markers such as p53, oestrogen receptor and p16 may be of use in problematic cases, but there is overlap and these may not therefore be of value in an individual case. It has been shown recently that high-mobility group AT-hook 2 (HMGA2) is expressed by most ovarian serous carcinomas, and our aim was to ascertain whether it is also expressed in uterine serous carcinoma and of value in its distinction from endometrioid carcinoma. Whole tissue sections of uterine serous (n = 33) and endometrioid (n = 38) carcinoma were immunostained using HMGA2 antibody. As many of the diagnostic problems relate to the distinction between serous carcinoma and grade 3 endometrioid carcinoma, tissue microarrays (TMAs) containing uterine serous (n = 71) and uterine grade 3 endometrioid (n = 68) carcinomas were also stained. Staining was classified as negative (totally negative or occasional nuclei positive), 1+ (<10% of nuclei positive), 2+ (10-49% of nuclei positive), 3+ (50-74% of nuclei positive), or 4+ (≥75% of nuclei positive). On the whole tissue sections, positive staining was also classified as weak, moderate, or strong, and an immunohistochemical composite score, taking into account both extent and intensity of staining, was calculated. On whole tissue sections, there was a statistically significant difference between HMGA2 staining in serous and endometrioid carcinomas with regard to both extent and composite score, with higher expression in serous carcinomas (P < 0.0001). Thirty of 33 (91%) serous carcinomas were positive, usually with diffuse (3+ or 4+) staining. All five cases of serous endometrial intraepithelial carcinoma (EIC) (the postulated precursor of uterine serous carcinoma) were positive, as were 14 of 38 (37%) endometrioid carcinomas, usually with 1+ or 2+ staining. There was a statistically significant difference in HMGA2 staining in the TMAs between the serous and grade 3 endometrioid carcinomas, with higher expression in the former (P < 0.0001). Immunoreactivity for HMGA2 is diffusely positive in whole tissue sections in most uterine serous carcinomas and negative in most endometrioid carcinomas, although, as with other markers, there is overlap in individual cases. In conjunction with other markers, HMGA2 may be of value in problematic uterine carcinomas where the differential diagnosis includes serous and endometrioid carcinoma. As HMGA2 is expressed in serous EIC, this suggests that it may be implicated in the early development of uterine serous carcinoma.


Release of plasminogen activator from normal and neoplastic endometrium.
In the medium of endometrial carcinoma cultures, anti-urokinase-reacting plasminogen activator was released in contrast to cultures of normal or hyperplastic endometrium.


Risk evaluation of adverse aortic events in patients with non-circular aortic annulus after transcatheter aortic valve implantation: a numerical study.
Transcatheter aortic valve implantation (TAVI) is a micro-invasive surgery used to treat patients with aortic stenosis (AS) efficiently. However, the uneven valve expansion can cause a non-circular annulus, which is one of the main factors leading to complications after TAVI. As a preliminary work, the main purpose of this study was to evaluate the risk of adverse aortic events in patients with a non-circular aortic annulus after TAVI. This study numerically investigated the distribution of four wall shear stress (WSS)-based indicators and three helicity-based indicators in eight patient-specific aortas with different annulus including circular, type I elliptical and type II elliptical shapes. Both elliptical annulus features can significantly enhance the intensity of the helicity (h2) in the ascending aorta (p < 0.001). However, for the type I elliptical annulus, the spiral flow structure was changed into low-velocity and disturbed flow pattern close to the inner side of the aortic arch. For the type II elliptical annulus, the spiral flow remained but became skewed in distribution. The elliptical annulus feature could increase the general level WSS-based indicators, especially in the ascending aorta. However, due to the disturbance of spiral flow or second helical flow in ascending aortas, areas with low TAWSS accompanied by high oscillatory shear index (OSI) and cross flow index (CFI) were observed in all the ascending aortas with non-circular annulus. The elliptical annulus feature can change the hemodynamic environment in the aortic arch, especially in the ascending aorta. Although both elliptical annulus features enhanced the strength of helicity, the uniform distribution of the helical flow was disturbed, especially in the ascending aorta, indicating the potential risk of adverse aortic events may increase. Therefore, for the patients without paravalvular leak but elliptical annulus shape after TAVI treatment, surgeons may be needed to consider further dilatation to make the non-circular annulus become circular.


[Dementia Care Manager for patients with dementia. Determination of the requirements and qualifications contents for nurses in the DelpHi-MV study].
Dementia is one of the most prevalent chronic progressive diseases in older age. The progression of dementia is associated with an increasing demand for patient care. Thus, the nursing profession fulfills important tasks in the supply of care in dementia. Care of dementia patients requires nurses with more specialized professional knowledge. Consequently, the development of new qualification concepts in dementia is needed. Therefore, the German Center for Neurodegenerative Diseases, Rostock/Greifswald, has developed a qualification according to the Dementia Care Management concept. A prospective cross-sectional study identified the tasks and qualifications of nurses as Dementia Care Managers. Overall, 27 tasks and 28 qualification items were identified for a nurse to qualify as a Dementia Care Manager. In the next step, the first version of the Dementia Care Management Curriculum was developed.


Identification of CYP443D1 (CYP74 clan) of Nematostella vectensis as a first cnidarian epoxyalcohol synthase and insights into its catalytic mechanism.
The CYP74 clan enzymes are responsible for the biosynthesis of numerous bioactive oxylipins in higher plants, some Proteobacteria, brown and green algae, and Metazoa. A novel putative CYP74 clan gene CYP443D1 of the starlet sea anemone (Nematostella vectensis, Cnidaria) has been cloned, and the properties of the corresponding recombinant protein have been studied in the present work. The recombinant CYP443D1 was incubated with the 9- and 13-hydroperoxides of linoleic and α-linolenic acids (9-HPOD, 13-HPOD, 9-HPOT, and 13-HPOT, respectively), as well as with the 9-hydroperoxide of γ-linolenic acid (γ-9-HPOT) and 15-hydroperoxide of eicosapentaenoic acid (15-HPEPE). The enzyme was active towards all C18-hydroperoxides with some preference to 9-HPOD. In contrast, 15-HPEPE was a poor substrate. The CYP443D1 specifically converted 9-HPOD into the oxiranyl carbinol 1, (9S,10R,11S,12Z)-9,10-epoxy-11-hydroxy-12-octadecenoic acid. Both 18O atoms from [18O2-hydroperoxy]9-HPOD were virtually quantitatively incorporated into product 1. Thus, the CYP443D1 exhibited epoxyalcohol synthase (EAS) activity. The 18O labelling data demonstrated that the reaction mechanism included three sequential steps: (1) hydroperoxyl homolysis, (2) oxy radical rearrangement into epoxyallylic radical, (3) hydroxyl rebound, resulting in oxiranyl carbinol formation. The 9-HPOT and γ-9-HPOT were also specifically converted into the oxiranyl carbinols, 15,16- and 6,7-dehydro analogues of compound 1, respectively. The 13-HPOD was converted into erythro- and threo-isomers of oxiranyl carbinol, as well as oxiranyl vinyl carbinols. The obtained results allow assignment of the name "N. vectensis EAS" (NvEAS) to CYP443D1. The NvEAS is a first EAS detected in Cnidaria.


A 3 year post-intervention follow-up on mortality in advanced heart failure (EVITA vitamin D supplementation trial).
Vitamin D supplementation is widely used in the clinical setting, but its effects on mortality and cardiovascular outcomes in patients with heart failure are unclear. This paper reports outcome data that were collected during follow-up of 3 years after closure of the EVITA trial (a 3 year randomized, placebo-controlled, intervention study with 4000 IU vitamin D daily in patients with advanced heart failure), to capture potential latency effects of vitamin D supplementation on clinical outcomes. The prespecified primary endpoint was overall mortality. Secondary endpoints included hospitalization, mechanical circulatory support implantation, high urgent listing for heart transplantation, and heart transplantation. For group comparisons, we used Cox regression models with a time-dependent categorical covariate. The calculated net difference in circulating 25-hydroxyvitamin D between the vitamin D and placebo groups dropped from 60.9 nmol/L at the end of the active study period to 3.2 nmol/L at the end of the post-intervention period. During the entire 6 year period, 73 patients (36.5%) died in the placebo group and 76 (38.8%) in the vitamin D group. Out of these 149 patients, 36 and 39 died during the first 3 years, and 37 and 37 during the second 3 years, respectively. The hazard ratio (HR) for mortality in the vitamin D versus the placebo group was 1.06 [95% confidence interval (CI): 0.68-1.66] for the first 3 years and 1.07 (95% CI: 0.68-1.70) for the 3 year post-intervention follow-up. Compared with the placebo group, the HRs for hospitalization and for mechanical circulatory support implant were significantly higher in the vitamin D group during vitamin D supplementation (HR = 1.31, 95% CI: 1.01-1.68 and HR = 2.01, 95% CI: 1.08-3.76, respectively) but not after vitamin D discontinuation (HR = 1.10, 95% CI: 0.62-1.94 and HR = 0.99, 95% CI: 0.38-2.56, respectively). There was no significant time-dependent effect on the risk of high urgent listing for heart transplantation and heart transplantation. No beneficial latency effects of vitamin D supplementation on overall mortality could be demonstrated. Instead, the disappearance of unfavourable findings in the vitamin D group (higher HRs for hospitalization and for mechanical circulatory support implant) after vitamin D discontinuation supports the assumption of adverse vitamin D effects on the cardiovascular system at doses of 4000 IU daily.


Does a history of cardiac disease or hypertension increase mortality following primary elective total hip arthroplasty?
Preoperative co-morbidities such as known coronary artery disease have commonly deemed a patient at 'high risk' for primary elective Total Hip Arthroplasty (THA). We prospectively collected data on 1744 patients who underwent primary elective THA between 1998 and 2004; 273 had a history of cardiac disease defined as a previous hospital admission with a diagnosis of angina pectoris or myocardial infarction; 594 patients had hypertension defined as that requiring treatment with antihypertensives. We also had data on preoperative age, sex and body mass index (BMI). There was no statistically significant increase in early mortality at three months with a history of cardiac disease or hypertension and this remained so when adjusting for the other factors in a multivariate analysis. Sex or BMI also did not have a statistically significant effect on the risk of death within three months. Increasing age was the only significant risk factor for early mortality (p<0.001). Longer-term mortality at two and five years in relation to these factors was also examined. Statistical analysis revealed that coronary history now showed a highly significant association (p<0.001) with long-term mortality in patients who survived more than three months. This remained significant (p=0.002) when adjusted for the other factors. Hypertension continued to have no effect, as did BMI. Age remained a significant risk factor and ASA was also a predictor of death, as has been previously shown. The overall long-term mortality following THAwas less than expected from the normal population, even in the subgroup with a coronary history. This study will assist clinicians when advising patients who have one of these common risk factors when seeking primary elective THA.


Characteristics of hepatitis B virus genotype G coinfected with genotype H in chimeric mice carrying human hepatocytes.
Accumulated evidence indicated that hepatitis B virus genotype G (HBV/G) is present exclusively in coinfection with other HBV genotypes. In Mexico, HBV/G from 6 men who had sex with men were coinfected with HBV/H. Phylogenetically complete genomes of the 6 Mexican HBV/G strains were closely related to previous ones from the US/Europe. Using uPA/SCID mice with human hepatocytes, monoinfection with HBV/G did not result in detectable HBV DNA in serum, whereas superinfection with HBV/G at week 10 inoculated HBV/H when HBV/H DNA was elevated to >10(7) copies/mL has enhanced the replication of HBV/G. The HBV/G was enhanced in another 3 inoculated with a serum passage containing HBV/G with a trace of HBV/H. Coinfection of mice with HBV/G and H induced fibrosis in the liver. In conclusion, the replication of HBV/G can be enhanced remarkably when it is coinfected with HBV/H. Coinfection with HBV/G may be directly cytopathic in immunosuppressive conditions.


Combination of a rapamycin analog (CCI-779) and interferon-gamma is more effective than single agents in treating a mouse model of tuberous sclerosis complex.
Tuberous sclerosis complex (TSC) is a familial tumor syndrome characterized by the development of hamartomas in the brain, heart, kidney, and skin. Disease-causing mutations in the TSC1 or TSC2 gene result in constitutive activation of the highly conserved mTOR signal transduction pathway, which regulates cell growth, proliferation, and metabolism. The mTOR inhibitor, rapamycin (sirolimus), reduces disease severity in rodent models of TSC, and is currently in phase II clinical trials. The cytokine interferon-gamma (IFN-gamma) is another potential therapeutic agent for TSC. A high-expressing IFN-gamma allele is associated with a lower frequency of kidney tumors in TSC patients, and treatment with exogenous IFN-gamma reduces the severity of TSC-related disease in mouse models. Here, we examine the effects of treating tumor-bearing nude mice with a combination of a rapamycin analog (CCI-779) and IFN-gamma. We observed that combination therapy was more effective than single agent therapy in reducing tumor growth and improving survival in this mouse model of TSC. Immunoblot and immunohistochemical analyses showed that tumors treated with CCI-779 plus IFN-gamma had decreased cell proliferation and increased cell death in comparison with untreated tumors or tumors treated with either agent alone. We also observed that CCI-779 resistance could develop with prolonged treatment. Taken together, our results show that targeting multiple cellular pathways is an effective strategy for treating TSC-related tumors, and underscore the importance of investigating combination therapy in future clinical trials for patients with TSC.


Percutaneous cholecystostomy as a definitive treatment for moderate and severe acute acalculous cholecystitis: a retrospective observational study.
In this study, we aimed to investigate risk factors for the relapse of moderate and severe acute acalculous cholecystitis (AAC) patients after initial percutaneous cholecystostomy (PC) and to identify the predictors of patient outcomes when choosing PC as a definitive treatment for AAC. The study population comprised 44 patients (median age 76 years; range 31-94 years) with moderate or severe AAC who underwent PC without subsequent cholecystectomy. According to the results of follow-up (followed for a median period of 17 months), the data of patients with recurrence versus no recurrence were compared. Patients were divided into the death and non-death groups based on patient status within 60 days after PC. Twenty-one (47.7%) had no recurrence of cholecystitis during the follow-up period after catheter removal (61-1348 days), six (13.6%) experienced recurrence of cholecystitis after PC, and 17 (38.6%) patients died during the indwelling tube period (5-60 days). The multivariate analysis showed that coronary heart disease (CHD) or congestive heart failure (odds ratio [OR] 26.50; 95% confidence interval [CI] 1.21-582.06; P = 0.038) was positively correlated with recurrence. The age-adjusted Charlson comorbidity index (OR 1.53; 95% CI 1.08-2.17; P = 0.018) was independently associated with 60-day mortality after PC. Our results suggest that CHD or congestive heart failure was an independent risk factor for relapse in moderate and severe AAC patients after initial PC. AAC patients with more comorbidities had worse outcomes.


ATM mutations that cause ataxia-telangiectasia are breast cancer susceptibility alleles.
We screened individuals from 443 familial breast cancer pedigrees and 521 controls for ATM sequence variants and identified 12 mutations in affected individuals and two in controls (P = 0.0047). The results demonstrate that ATM mutations that cause ataxia-telangiectasia in biallelic carriers are breast cancer susceptibility alleles in monoallelic carriers, with an estimated relative risk of 2.37 (95% confidence interval (c.i.) = 1.51-3.78, P = 0.0003). There was no evidence that other classes of ATM variant confer a risk of breast cancer.


Patients who seek a second opinion: are they different from the typical referral?
Little is known about what factors determine when a patient decides to seek a second opinion from an internist. The Canadian health care system, which places no responsibility for payment on the patient, allows a unique opportunity to assess such factors when financial influences are minimized. We hypothesized that in such a milieu patients with functional diseases might be more apt to seek a second opinion. In a university-based gastrointestinal unit we investigated how many of the patients, referred for the first time, were seeking a second opinion--i.e., a second consultation within 2 years of seeing a gastroenterologist. We compared the characteristics of 20 patients with 246 patients referred for the first time to a university-based gastrointestinal unit. All patients were interviewed at clinic registration, prior to seeing the gastroenterologist. Variables assessed included demographic characteristics, health care utilization within the last year, and perceived health status. A second questionnaire was offered to second opinion patients to determine (1) reasons for seeking a second opinion, and (2) whose decision it was to seek the second opinion. Following the interview, the patient's referring physician was contacted and asked identical questions. Both referring physician and gastroenterologist assessed every patient on a functional rating (FR) scale of 1 to 5 (with 1 being definitely organic and 5 being definitely functional).(ABSTRACT TRUNCATED AT 250 WORDS)


Religiosity, health, and well-being among Kuwaiti personnel.
In a sample of 424 Kuwaiti personnel (219 men, 205 women; M age = 37.6 yr., SD = 8.9; M age = 33.4 yr., SD = 7.9, respectively), self-ratings of religiosity were significantly and positively correlated with the self-ratings of physical health, mental health, and happiness, as well as the Oxford Happiness Inventory, the Love of Life Scale, and the Satisfaction with Life Scale among men and women. Principal components analysis of the correlation matrix yielded only one salient factor labeled "Well-being, health and religiosity" that explained 52.7% and 56.5% of the variance for men and women, respectively. Religiosity is an important element in the lives of the majority of the present sample of Kuwaiti Muslim employees.


Coronary thrombosis after European adder bite in a patient on dual antiplatelet therapy: A case report.
Until now very few cases of an adverse cardiovascular event have been described following European viper envenomation (Aravanis et al., 1982) (Aravanis et al., 1982) (Aravanis et al., 1982) (Aravanis et al., 1982). In fact, cardiac toxicity following snake bite is rare and primary reported from tropical and subtropical areas with only twenty-one cases of myocardial infarction reported in literature. Herein, we report a case of European viper envenomation associated with coronary artery thrombosis complicated by acute myocardial infarction and cardiac arrest. A 62-year-old man, with a history of cardiovascular disease, on dual antiplatelet therapy with ticagrelor and acetylsalicylic acid, was admitted to the Emergency Department, after a bite, on the right hand, from a snake recognized by a herpetologist as a Vipera aspis francisciredi. At ED presentation, 2 hours after the bite, he manifested with vomiting, hypotension (90/60 mmHg) and mild oedema at the bite site. Standard electrocardiogram and troponin were normal at admission. One hour after the admission the patient developed cardiocirculatory arrest (ACC) with return of spontaneous circulation (ROSC) after cardiopolmunary resuscitation. Post ROSC-ECG showed an ST-elevation on anteroseptal and lateral leads and 1-vial of Viper Venom Antitoxin (Biomed®) was i.v. administered. During the next 3 hours three other episodes of ACC occurred, always with restoration of spontaneous circulation. Percutaneous transluminal coronary angiography showed a thrombus on the bifurcation of anterior descending coronary artery and diagonal 1 without an underlined atherosclerotic plaque. Neurologic clinical manifestations also occurred 12 hours after the bite: bilateral ptosis and facial paresthesia and a second vial of the same viper antivenom administered. The patient was discharged after 9 days of hospitalization without sequelae. Our case show that cardiotoxicity is a rare but possible event after snake envenomation in Europe, even if with mechanisms remains to be studied. Vipera aspis has been known to cause primarily neurotoxic manifestations, but a coagulation factor X activator have also been isolated from its venom. Moreover, a specific serine peptidase that can target both PAR1 and PAR3, that are responsible for alternate pathways of platelet aggregation, have been characterized in the venom of a viper. Coronary thrombosis in our case could thus be secondary to a combination of prothrombotic systemic state and platelets dysfunction, in a patient with predisposing factors. Antivenom specific antidotal therapy role in preventing cardiotoxicity still need to be elucidated, but it remains the mainstay of treatment together with coronary angiography if necessary.


Measuring Protein-Protein Interactions in Cells using Nanoluciferase Bioluminescence Resonance Energy Transfer (NanoBRET) Assay.
Protein-protein interactions (PPIs) are increasingly recognized for their roles in functional cellular networks and their importance in disease-targeting contexts. Assessing PPI in the native cellular environment is challenging and requires specific and quantitative methods. Bioluminescence resonance energy transfer (BRET) is a biophysical process that can be used to quantify PPI. With Nanoluciferase bioluminescent protein as a donor and a fluorescent chloroalkane ligand covalently bound to HaloTag protein as an acceptor, NanoBRET provides a versatile and robust system to quantitatively measure PPI in living cells. BRET efficiency is proportional to the distance between the donor and acceptor, allowing for the measurement of PPI in real time. In this paper, we describe the use of NanoBRET to study specific interactions between proteins of interest in living cells that can be perturbed by using small-molecule antagonists and genetic mutations. Here, we provide a detailed protocol for expressing NanoLuc and HaloTag fusion proteins in cell culture and the necessary optimization of NanoBRET assay conditions. Our example results demonstrate the reliability and sensitivity of NanoBRET for measuring interactions between proteins, protein domains, and short peptides and quantitating the PPI antagonist compound activity in living cells.


Lipid peroxidation in Rheumatoid arthritis; consequences and monitoring.
The epidemiological studies have shown that rheumatoid arthritis (RA) leads to oxidative stress formation. Therefore the aim of this study has been to estimate the lipid peroxidation in RA patients. Moreover the reasons and consequences of changes in lipid peroxidation were estimated. 73 patients with RA and 62 healthy subjects were included into the study. Lipid peroxidation was estimated by the measurement of phospholipid arachidonic acid (AA) and linoleic acid (LA) as well as aldehydes: 4-hydroxynonenal (4-HNE), 4-hydroxyhexenal (4-HHE), malondialdehyde (MDA), acrolein, crotonaldehyde, and 4-oxononenal (4-ONE) that were determined by GC and GCMS, while 8-isoprostanes (8-isoPGF2a) - was determined by LCMS. Phospholipase A2 (PLA2), platelet-activating factor acetylhydrolase (PAF-AH) and glutathione peroxidase (GSH-Px) activity were determined spectrophotometrically, while vitamin E was determined by HPLC. Plasma of RA patients was characterized by higher vitamin E level and decreased GSH-Px activity as well as the level of phospholipid AA and LA compared to healthy subjects. The level of all examined aldehydes was significantly increased in plasma patients but higher increase was observed in urine of RA patients in comparison with control group. The 8-isoprostanes level was approximately 9-fold higher in the plasma and 3-fold higher in the urine of RA patients compared to healthy subjects, but in the urine the changes in 8-isoprostanes level was correlated with RA progression. A significant increase in 4-HNE-modified protein adducts level as well as a decrease in the activity of PAF-AH and PLA2 were observed in the plasma of RA patients. The level of lipid peroxidation products in the plasma and the urine may be the indicator of RA, but additionally urine 8-isoprostanes may be the useful and reliable biomarker of RA progression.


Transcriptional regulation of the hepatocyte growth factor gene by pyrrolidine dithiocarbamate.
Hepatocyte growth factor (HGF) mediates cancer cell invasion and metastasis. This study characterised the down-regulation of HGF expression by pyrrolidine dithiocarbamate (PDTC), which markedly reduced HGF mRNA expression and protein production in MRC-5 cells. Reporter gene studies revealed that PDTC inhibited HGF gene transcription and that the response element is located in the region -75 to +42 bp flanking the transcription initiation site. Electrophoretic mobility shift assay identified three specific protein complexes binding in this region, which were abrogated by exposure of cells to PDTC. PDTC deserves further investigation as a novel therapeutic agent for HGF-driven cancers.


Clinical outcome of older adults with acute myeloid Leukemia: An analysis of a large tertiary referral Center over two decades.
In older adults with acute myeloid leukemia (AML), the overall outcome is still dismal and long-term data on survival are scarce, particularly outside of clinical trials. Here, we assess characteristics, prognostic factors and long-term survival in patients ≥60 years who were treated for AML at our center over the past 17 years. 590 older adults with newly diagnosed AML were characterized according to Eastern Cooperative Oncology Group (ECOG) score, Charlson comorbidity index (CCI), European LeukemiaNet (ELN) risk, type of therapy, serum ferritin (SF) and further baseline characteristics. Survival analysis was performed accordingly. Median age was 68 years and most patients were in good general condition. Median follow-up was 55.8 months. Of all patients, 66% received intensive chemotherapy (IC) +/- allogeneic hematopoietic stem cell transplantation (allo-HSCT). The remaining cohort received palliative chemotherapy (PC, 26%) or best supportive care only (BSC, 8%). Enrollment rate for interventional clinical trials was 26%. 5-year overall survival (OS) and relapse-free survival (RFS) were 18% (median 12.5 months) and 11,5% (median 10.0 months). Long-term survival was independently influenced by ECOG score, ELN risk group, baseline SF, previous myocardial infarction, and choice of therapy, but not consistently by age or CCI. Considering therapeutic subgroups, the contribution of particular parameters in predicting OS was most compelling in IC patients, but less consistent with PC or BSC. Our results provide thorough insights into prognostication within therapeutic subgroups and emphasize the need for more detailed prognostic algorithms and routine geriatric assessment in the treatment of older adults with AML.


A bioabsorbable everolimus-eluting coronary stent system for patients with single de-novo coronary artery lesions (ABSORB): a prospective open-label trial.
A fully bioabsorbable drug-eluting coronary stent that scaffolds the vessel wall when needed and then disappears once the acute recoil and constrictive remodelling processes have subsided has theoretical advantages. The bioasorbable everolimus-eluting stent (BVS) has a backbone of poly-L-lactic acid that provides the support and a coating of poly-D,L-lactic acid that contains and controls the release of the antiproliferative agent everolimus. We assessed the feasibility and safety of this BVS stent. In this prospective, open-label study we enrolled 30 patients who had either stable, unstable, or silent ischaemia and a single de-novo lesion that was suitable for treatment with a single 3.0 x 12 mm or 3.0 x 18 mm stent. Patients were enrolled from four academic hospitals in Auckland, Rotterdam, Krakow, and Skejby. The composite endpoint was cardiac death, myocardial infarction, and ischaemia-driven target lesion revascularisation. Angiographic endpoints were available for 26 patients and intravascular-ultrasound endpoints for 24 patients. Clinical endpoints were assessed in all 30 patients at 6 and 12 months. In a subset of 13 patients, optical coherence tomography was undertaken at baseline and follow-up. Analysis was by intention to treat. This study is registered with ClinicalTrials.gov, number NCT00300131. Procedural success was 100% (30/30 patients), and device success 94% (29/31 attempts at implantation of the stent). At 1 year, the rate of major adverse cardiac events was 3.3%, with only one patient having a non-Q wave myocardial infarction and no target lesion revascularisations. No late stent thromboses were recorded. At 6-month follow-up, the angiographic in-stent late loss was 0.44 (0.35) mm and was mainly due to a mild reduction of the stent area (-11.8%) as measured by intravascular ultrasound. The neointimal area was small (0.30 [SD 0.44] mm2), with a minimal area obstruction of 5.5%. This study shows the feasibility of implantation of the bioabsorbable everolimus-eluting stent, with an acceptable in-stent late loss, minimal intrastent neointimal hyperplasia, and a low stent area obstruction. Abbott Vascular.


Effect of dietary fiber on colonic bacterial enzymes and bile acids in relation to colon cancer.
Because of the potential significance of colonic bacteria and secondary bile acids in the pathogenesis of colon cancer, the present study investigated the effect of different types of dietary fiber on fecal bacterial enzymes, namely, beta-glucuronidase, 7 alpha-dehydroxylase, nitroreductase, and azoreductase, and on bile acids and neutral sterols in premenopausal women. The subjects consumed 13-15 g of wheat, oat, or corn bran daily for 8 weeks in addition to their normal diet. Stools collected during the normal and fiber diet periods were analyzed for the above constituents. Dietary wheat bran decreased the concentrations of fecal deoxycholic acid, lithocholic acid, 12-ketolithocholic acid, and neutral sterols and the activities of all bacterial enzymes. Oat bran had no effect on secondary bile acids and 7 alpha-dehydroxylase but decreased beta-glucuronidase, nitroreductase, and azoreductase levels. Dietary corn bran increased 7 alpha-dehydroxylase, lithocholic acid, and cholesterol levels and decreased deoxycholic acid coprostanol, cholestenone, nitroreductase, and azoreductase levels. These results show that the modifying effect of dietary fiber on secondary bile acids and bacterial enzymes that may play a role in carcinogenesis depends on the type of fiber consumed.


Differences in foliage quality of young jack pine (Pinus banksiana Lamb.) on burned and clearcut sites: effects on jack pine budworm (Choristoneura pinus pinus Freeman).
Suitability of young jack pine as a host for jack pine budworm was examined on similarly-aged trees growing on two areas previously burned in wildfires and on two previously clearcut areas in northwest Wisconsin. Nitrogen, monoterpenes, and moisture levels of foliage, and xylem water potential were measured and related to larval survival and pupal weight of caged jack pine budworm larvae. Nitrogen, monoterpenes, needle weight, and needle moisture were higher in trees growing on clearcut sites than on burned area trees. Surival of budworms to early and late instar, pupation, and adult eclosion was greater for larvae caged on clearcut-area trees than on burned-area trees. Female pupal weight differed between older (ca 10 years old) and younger (ca 8 years old) trees, but not between clearcut and burned areas. Mean female pupal weight was greatest on lownitrogen trees, where larval survival was lowest. Foliar nitrogen was consistently included as a significant predictor in budworm survival regressions. Regressions indicated larval survival and pupal weight may be associated with different tree- and foliage-related traits. Results suggest long-lasting effects of previous forest disturbance may subsequently affect herbivorous insects such as jack pine budworm.


An Insight into the Hepatoprotective Activity and Structure-activity Relationships of Flavonoids.
Flavonoids are a class of polyphenolic bioactive compounds obtained from plants, which have a wide range of chemical structures and properties. More than 9000 distinct flavonoid molecules have been identified and have been found to regulate numerous developmental processes and play key biological roles in living organisms. This review aims to highlight the hepatoprotective potentiality of flavonoids and corelate their pharmacological activity with their chemical structure. With the advancement in the field of research related to phytochemicals, it is evident that flavonoids have versatile health benefits, viz., antioxidant property, free radical scavenging capacity, anticancer activity. The basic structures are C6-C3-C6 rings with various substitution patterns, resulting in a succession of subclass compounds, and the relationships between chemical structures and bioactivity have previously been investigated. The hepatoprotective effects of bioactive flavonoids derived from plants have been widely linked to their antioxidant activity, antiinflammatory activity, effects on Sterol Regulatory Element- binding Proteins (SREBP), Peroxisome Proliferator-activated Receptor gamma (PPARγ) receptors, and inflammatory mediator cytokines according to numerous studies. The C2-C3 double bond at the A ring, as well as the hydroxyl groups of C3'or C4', and the carbonyl group at position C4, have been shown to augment their hepatoprotective activities; however, hydroxymethylation at C3' and C4' has been found to diminish the hepatoprotective activity. The impact of flavonoid moieties and the structure-activity relationship of flavonoids related to combating various hepatic disorders have been vividly discussed in this review paper.


[Toric IOL's].
Toric intraocular lenses have been increasingly utilized in cataract surgery and are considered "premium lenses" They provide a reliable and effective option for patients with astigmatism undergoing cataract surgery. We present various methods for the correction of astigmatism during cataract surgery. We present the available models of pseudophakic toric IOL's, including monofocal and multifocal lenses. We present indications for toric IOL's in cases of regular and irregular astigmatism.


Osteogenic potential of murine periosteum for critical-size cranial defects.
Tissue engineering of bone has combined bespoke scaffolds and osteoinductive factors to maintain functional osteoprogenitor cells, and the periosteum has been confirmed as a satisfactory source of osteoblasts. Suitable matrices have been identified that support cell proliferation and differentiation, including demineralised bone matrix (both compatible and osteoinductive) and acellular human dermis. We have evaluated the osteogenic potential of an osteogenic unit, developed by combining periosteum, demineralised bone matrix, and acellular human dermis, in rodents with critical-size cranial defects. Briefly, remnants from the superior maxillary periosteum were used to harvest cells, which were characterised by flow cytometry and reverse retrotranscriptase-polymerase chain reaction (RT-PCR). Cells were cultured into the osteogenic unit and assessed for viability before being implanted into 3 rodents, These were compared with the control group (n=3) after three months. Histological analyses were made after staining with haematoxylin and eosin and Von Kossa, and immunostaining, and confirmed viable cells that stained for CD90, CD73, CD166, runt-related transcription factor, osteopontin, and collagen type I in the experimental group, while in the control group there was only connective tissue on the edges of the bone in the injury zone. We conclude that osteogenic unit constructs have the osteogenic and regenerative potential for use in engineering bone tissue.


Breast Cancer Management During COVID-19 Pandemic in Madrid: Surgical Strategy.
From the first case of SARS-CoV-2 infection in Wuhan (China), the infection spread all around the world causing a pandemic of coronavirus disease-2019 (COVID-19). Spain has been one of the most severely affected countries, and Madrid has reported a high number of cases and deaths. We discuss our strategies for optimal breast cancer management during COVID-19 pandemic. This was a retrospective observational study at Clínico San Carlos Hospital to analyze the management of patients with breast cancer during the pandemic outbreak and the surgical strategy after the pandemic outbreak. We created a practical and dynamic tool based on a "traffic light" system for prioritizing surgical time. Every patient was contacted by telephone with a preoperative COVID-19 protocol. After surgical procedures, patient satisfaction was assessed using the European Organisation for Research and Treatment of Cancer in-patient satisfaction with cancer care questionnaire (EORTC IN-PATSAT32). Patients with breast cancer actively treated with surgical procedures were put on a waiting list and received systemic therapy. Telemedicine was used to evaluate any side effects and to avoid unnecessary hospital visits. Surgery was only considered after the pandemic outbreak, and then, only those procedures designed to minimize surgical complications and, therefore, reduce hospital stay. We also measured patients' satisfaction with medical and nursing scales that resulted in a "very good" evaluation tending to "excellent". It is necessary to adapt management of oncology treatment and surgical strategy to optimize resources during the COVID-19 pandemic. Patients' perception of care quality and the degree of patients' satisfaction with health services has potential relevance in the absence of outcome data.


Regional specialization in pyramidal cell structure in the limbic cortex of the vervet monkey (Cercopithecus pygerythrus): an intracellular injection study of the anterior and posterior cingulate gyrus.
The pyramidal cell phenotype varies quite dramatically in structure among different cortical areas in the primate brain. Comparative studies in visual cortex, in particular, but also in sensorimotor and prefrontal cortex, reveal systematic trends for pyramidal cell specialization in functionally related cortical areas. Moreover, there are systematic differences in the extent of these trends between different primate species. Recently we demonstrated differences in pyramidal cell structure in the cingulate cortex of the macaque monkey; however, in the absence of other comparative data it remains unknown as to whether the neuronal phenotype differs in cingulate cortex between species. Here we extend the basis for comparison by studying the structure of the basal dendritic trees of layer III pyramidal cells in the posterior and anterior cingulate gyrus of the vervet monkey (Brodmann's areas 23 and 24, respectively). Cells were injected with Lucifer Yellow in flat-mounted cortical slices, and processed for a light-stable DAB reaction product. Size, branching pattern, and spine density of basal dendritic arbors were determined, and somal areas measured. As in the macaque monkey, we found that pyramidal cells in anterior cingulate gyrus (area 24) were more branched and more spinous than those in posterior cingulate gyrus (area 23). In addition, the extent of the difference in pyramidal cell structure between these two cortical regions was less in the vervet monkey than in the macaque monkey.


Postoperative acute cholecystitis complicating urologic operations.
Two cases are described of acute gangrenous cholecystitis following urologic operations. The subject is reviewed. The predominance of urologic procedures as the preceding factor for this postoperative complication is pointed out. The possible causes, the presenting features, the diagnosis, management, and serious prognosis are discussed. Awareness of the condition, early diagnosis, and operation are recommended. Its importance to the urologist is stressed.


Phospholipid methylation in canine myocardium: kinetic characteristics and the effect of endotoxin administration.
The kinetic characteristics and the effect of endotoxin administration on the enzymatic methylation of phospholipids in dog heart microsomes were studied using S-adenosyl-L-[methyl-3H]methionine as a methyl donor. Kinetic studies in control dogs reveal that the stepwise methylation of phosphatidylethanolamine to phosphatidylcholine was catalyzed by three different enzymes. Methyltransferase I catalyzed the methylation of phosphatidylethanolamine to phosphatidyl-N-monomethylethanolamine, had a very low Km (approximately 1.5 microM) for S-adenosylmethionine, and a pH optimum of 6.5, and it was stimulated by Mg2+ and Ca2+. Methyltransferase II catalyzed the methylation of phosphatidyl-N-monomethylethanolamine to phosphatidyl-N,N-dimethylethanolamine, had a low Km (8-12 microM) for S-adenosylmethionine, and a pH optimum of 8.5, and it was stimulated by low concentrations (less than 1 mM) of Ca2+ but was unaffected by Mg2+. Methyltransferase III catalyzed the formation of phosphatidylcholine from phosphatidyl-N,N-dimethylethanolamine, had a high Km (approximately 33 microM) for S-adenosylmethionine, and a pH optimum of 9.5, and it was unaffected by Mg2+ or Ca2+. Experiments with trypsin digestion indicate that methyltransferases I and III were partially embedded while methyltransferase II was completely exposed to the surface of the membrane. Endotoxin administration (2 and 4 hr) decreased the Km and Vmax by 30 to 36% and 24 to 37.7%, respectively, for S-adenosylmethionine. Since the enzymatic methylation of phospholipids has been implicated to play an important role in the regulation of membrane structure and function, the endotoxin-induced decreases in the Km and Vmax of phospholipid-methylating enzymes in dog heart microsomes may contribute to the development of myocardial dysfunction in endotoxin shock.


Bilateral autoimmune optic neuritis and vitreitis related to CRMP-5-IgG: intravitreal triamcinolone acetonide therapy of four eyes.
To report the vision outcome following intravitreal triamcinolone acetonide (IVTA) as adjunctive therapy in four eyes of two patients with paraneoplastic autoimmune optic neuritis and vitreitis related to CRMP (Collapsin-Response-Mediator Protein)-5-IgG. Retrospective case series.MethodsChart review of four eyes. Preoperative visions were: patient 1, 20/50 OD, 20/25 OS; patient 2, counting fingers (CF) at two feet OD, and CF at three feet OS. At last follow-up, the postoperative visions were 20/40, 20/50 and 20/200, 20/60, respectively. All signs of optic disc swelling and vitreitis had abated. Use of IVTAin paraneoplastic autoimmune optic neuritis and vitreitis related to CRMP-5-IgG was followed by a marked decrease in inflammation and stabilization or improvement of vision. These observations support this form of adjunctive therapy in patients whose intraocular pathology is attributed to paraneoplastic autoimmunity.


Ultrashort time-to-echo T2* and T2* relaxometry for evaluation of lumbar disc degeneration: a comparative study.
To compare potential of ultrashort time-to-echo (UTE) T2* mapping and T2* values from T2*-weighted imaging for assessing lumbar intervertebral disc degeneration (IVDD),with Pfirrmann grading as a reference standard. UTE-T2* and T2* values of 366 lumbar discs (L1/2-L5/S1) in 76 subjects were measured in 3 segmented regions: anterior annulus fibrosus, nucleus pulposus (NP), and posterior annulus fibrosus. Lumbar intervertebral discs were divided into 3 categories based on 5-level Pfirrmann grading: normal (Pfirrmann grade I),early disc degeneration (Pfirrmann grades II-III), and advanced disc degeneration (Pfirrmann grades IV-V). Regional differences between UTE-T2* and T2* relaxometry and correlation with degeneration were statistically analyzed. UTE-T2* and T2*value correlated negatively with Pfirrmann grades (P < 0.001). In NP, correlations with Pfirrmann grade were high with UTE-T2* values (r = - 0.733; P < 0.001) and moderate with T2* values (r = -0.654; P < 0.001). Diagnostic accuracy of detecting early IVDD was better with UTE-T2* mapping than T2* mapping (P < 0.05),with receiver operating characteristic analysis area under the curve of 0.715-0.876. UTE-T2* relaxometry provides another promising magnetic resonance imaging sequence for quantitatively evaluate lumbar IVDD and was more accurate than T2*mapping in the earlier stage degenerative process.


Adsorption and isotope effects by ion exchange with 1-aza-12-crown-4 bonded Merrifield peptide resin.
Magnesium isotope effects were investigated by chemical ion exchange with synthesized 1-aza-12-crown-4 bonded Merrifield peptide resin using elution chromatography. The capacity of azacrown ion exchanger was 0.89 meq/g dry resin. The heavier isotopes of magnesium were enriched in the resin phase, while the lighter isotopes were enriched in the solution phase. The hydration effect is less than the complexation and isotope mass effects. The single stage separation factor was determined according to the method of Glueckauf from the elution curve and isotopic assays. The separation factors of 24Mg(2+)-25Mg(2+), 24Mg(2+)-26Mg(2+), and 25Mg(2+)-26Mg(2+) were 1.012, 1.023, and 1.011, respectively.


Drug-Drug Interaction of Letermovir and Atorvastatin in Healthy Participants.
Letermovir (MK-8228/AIC246) is a cytomegalovirus (CMV) DNA terminase complex inhibitor for CMV prophylaxis in adult patients undergoing hematopoietic stem cell transplant. It is cytochrome P450 (CYP) 3A inhibitor and inhibits organic anion transporting polypeptide 1B1/3 and breast cancer resistance protein transporters. Atorvastatin (ATV), a commonly used treatment for hypercholesterolemia, is a substrate of organic anion transporting polypeptide 1B1, potentially breast cancer resistance protein, and CYP3A. As letermovir may be coadministered with ATV, the effect of multiple-dose letermovir 480 mg once daily on the pharmacokinetics of single-dose ATV 20 mg and its metabolites (ortho-hydroxyatorvastatin [o-OH-ATV] and para-hydroxyatorvastatin [p-OH-ATV]) was evaluated in an open-label trial in healthy female adults (N = 14). ATV area under the plasma concentration-time curve from time 0 to infinity and maximum plasma concentration (Cmax ) increased ≈3-fold with letermovir coadministration. The time to ATV Cmax also increased, while apparent clearance decreased. The exposures of o-OH-ATV and p-OH-ATV were comparable in the presence versus absence of letermovir; however, o-OH-ATV Cmax decreased by 60% with coadministration, while p-OH-ATV Cmax was similar. Due to the increase in ATV exposure with letermovir coadministration, statin-associated adverse events such as myopathy should be closely monitored following coadministration. The dose of ATV should not exceed 20 mg daily when coadministered with letermovir.


Stem cell therapy is a promising tool for refractory angina: a meta-analysis of randomized controlled trials.
So far, relatively few studies have addressed the use of stem cells to treat patients with refractory angina. Moreover, the results of current studies were discrepant. The objective of this meta-analysis was to evaluate the efficacy and safety of this treatment on a relatively large scale. Studies were identified through PubMed, CENTRAL, EMBASE, reviews, and reference lists of relevant papers. The weighted mean difference was calculated with random-effect models for net changes in exercise tolerance and angina frequency, and odds ratio (OR) with fixed-effect models for myocardial infarction (MI) and death. Five randomized controlled trials, with a total of 381 patients, were included in the analysis. Compared with the controls, patients who received stem cell therapy had a significant improvement in exercise tolerance of 61.3 seconds (95% confidence interval [CI], 18.1-104.4; P = 0.005; I(2) = 58%); an obvious reduction in angina frequency of 7.3 episodes per week (95% CI, -13.4 to -1.2; P = 0.02; I(2) = 93%); and lower risk of MI, with an OR of 0.37 (95% CI, 0.14-0.95; P = 0.04; I(2) = 0%). No difference was detected for the risk of death (OR, 0.33; 95% CI, 0.08-1.39; P = 0.13; I(2) = 20%). Stem cell therapy appears to be effective and safe in the management of patients with refractory angina. The findings need confirmation in larger-scale studies with longer follow-up.


The experimental and numerical study of indirect effect of a rifle bullet on the bone.
We study the transient indirect effect of a rifle bullet on bone in the gelatin-bone composite target experimentally and computationally. The process of a 56 type 7.62-mm rifle bullet penetrating the composite target has been simulated using numerical method. The experiment provided the criteria for verifying the correctness of the numerical model. We have obtained tomographic data of bone by CT scans, and also defined the bone as different layers by the gray scale to simulate its heterogeneity. The computed results are in good agreement with the experimental data. Effects of the impact velocity and bone location on damage caused to the composite target have also been studied. The numerical results imply the follows: When the velocity of bullet increases, the stress on bone also increases with the earlier pressure peak; When the bone is located in a certain distance from the trajectory, it will not be fractured, although it is affected by the stress wave.


Characterization of BSA unfolding and aggregation using a single-capillary viscometer and dynamic surface tension detector.
A dynamic surface tension detector (DSTD) has been equipped with an additional pressure sensor for simultaneous viscosity measurements, as a detector for flow injection analysis. The viscosity measurement is based on a single capillary viscometer (SCV) placed in parallel configuration with the DSTD. The viscometer in the optimized conditions consists of a PEEK capillary (i.d.=0.25 mm, L=75 cm) kept at constant temperature using a thermostatic bath, which leads on the two sides to the two arms of a differential piezoelectric pressure transducer with a range of 0-35 psi. The DSTD, described previously, measures the changing pressure across the liquid/air interface of 2 μL drops repeatedly forming at the end of a capillary. SCV performance has been evaluated by measuring dynamic viscosity of water/glycerol mixtures analysed in flow injection and comparing the results with the values reported in the literature. The detection limits of SCV and DSTD, calculated as 3σ of the blank, were 0.012 cP and 0.6 dyn cm(-1), respectively. The FI-SCV-DSTD system has been applied to the study of temperature-induced denaturation/aggregation process in bovine serum albumin (BSA). The results have been supported and discussed with respect to BSA conformational analysis performed using Fourier Transform infrared spectroscopy.


Boosting the photocatalytic activity and stability of Cu2O for CO2 conversion by LaTiO2N.
Cu2O is one of the most promising photocatalysts for visible light driven CO2 conversion, but its practical application in artificial photosynthesis is challenged to severe photocorrosion. Herein, a Cu2O/LaTiO2N composite is successfully synthesized by loading Cu2O on LaTiO2N through an ascorbic acid reduction method, which significantly improves the stability and activity of Cu2O for visible light driven photocatalytic CO2 conversion, and shows higher photocatalytic performance than most Cu2O-based photocatalysts reported. The superior photocatalytic performance is attributed to the formation of type-II heterojunction between Cu2O and LaTiO2N, which facilitates the separation and transfer of photogenerated electrons and holes. This work provides an effective strategy for the construction of stable and efficient Cu2O-based photocatalysts.


[Investigations on the clothing of preschool-children during winter (author's transl)].
These investigations should control the effect of increased indoor-temperatures in facilities for children and flats on the clothing-habits of 1--3 aged children during the heating period. The indoor- and outdoor-clothing was scored by the number of dress-layers investigated in 977 crèche-children. All these attempts were carried out while 23 days during the winter 1976/77 in 2 crèches which differed significantly referring to the distribution of indoor-temperature. The statistical evaluation results that all children were dressed nearly in the like wise in spite of a considerable difference of the temperature in both crèches; only the covering of the arms was significant less in the warmer new-builded crèch. The investigations showed further that the outdoor-clothing of the children was often not according to the climate. Even 10 degrees C difference of temperature were rarely followed by variations of the outdoor-clothing. Finally the dress of 1--3 aged children can be estimated as too abundant.


Intensity modulated radiation therapy: film verification of planar dose maps.
A new slow response radiographic film (Kodak EDR) is compared with a more traditional faster responding dosimetry film (Kodak XV) in an intensity modulated radiation therapy x-ray field. Dose profiles derived from the two films are compared with doses obtained using a radiotherapy dose planning system (Philips-Pinnacle) which calculates planar dose using a collapsed cone convolution algorithm. Comparisons of the dose maps delivered from film with the Pinnacle dose maps are useful to ensure accurate dose delivery. The Pinnacle dose maps agreed with measurement using both film types to within +/- 3% (in the umbral region) at depths ranging from d(max) to 15 cm. Both XV and EDR films can be used to verify IMRT. EDR film is better suited for dosimetry for combined field dose maps due to its useful dose range of 1-5 Gy. We found XV to be more suitable for individual field dose maps (dose range 0-1 Gy) as no scaling of monitor units were required to achieve acceptable optical density response.


De Novo Mutations in Patients with Ataxic CP.
As a part of a large study investigating childhood ataxias in the UK and Switzerland, Schnekenberg et al. analyzed the genetic associations with congenital cerebellar ataxia in 10 patients using either a targeted next generation sequencing panel of 118 genes or trio-based exome sequencing.


[Changes in the degree of sensitivity to antibiotics of Shigella isolated from patients with dysentery in a city infectious disease hospital over the past 8 years].
Changes in sensitivity of Shigella and in particular Shigella flexneri isolated from dysentery patients in a city infection hospital within 8 years i.e. from 1977 to 1985 were followed. It was shown that the changes were to the greater extent connected with preferable use of certain antibiotics in the hospital.


Loss of heterozygosity and point mutation at Aprt locus in T cells and fibroblasts of Pms2-/- mice.
Mice null for the Pms2 mismatch repair (MMR) gene exhibit a predisposition to lymphoma, microsatellite repeat instability, and failure of spermatogenesis. To study the role of Pms2 in the maintenance of in vivo genomic integrity in somatic cells, we characterized Aprt mutations in T cells and fibroblasts of 129 x C3H Pms2-/-Aprt+/- mice. The spontaneous frequency of DAP-resistant T lymphocytes, as a consequence of APRT-deficiency, was increased threefold. Point mutation, which accounted for less than 20% of the DAP(r) mutant clones in Pms2+/+ mice, was predominant in the mutant T cell clones from Pms2-/- mice. These point mutations were predominantly TA to CG transitions. Fibroblasts of Pms2-/- mice exhibited only a modest increase in the frequency of clones with point mutations, such that mitotic recombination was still the primary cause of APRT deficiency. Thus, the mutator phenotype as a consequence of PMS2 deficiency is tissue-dependent, which may be related to the tissue-specific tumor proneness of Pms2-/- mice.


Congenital peritoneal mesothelioma.
Only three primary peritoneal mesotheliomas in children have been previously reported. We describe a 6-wk-old girl with a papillary peritoneal mesothelioma of low grade malignancy. This is probably a congenital mesothelioma and is the youngest patient reported to date.


Meningiomas: Preoperative predictive histopathological grading based on radiomics of MRI.
We aimed to develop a radiomics model to predict the histopathological grading of meningiomas by magnetic resonance imaging (MRI) before surgery. We recruited 131 patients with pathological diagnosis of meningiomas. All the patients had undergone MRI before surgery on a 3.0 T MRI scanner to obtain T1 fluid- attenuated inversion recovery (T1 FLAIR) images, T2-weighted images (T2WI) and T1 FLAIR with contrast enhancement (CE-T1 FLAIR) images covering the whole brain. The removing features with low variance, univariate feature selection, and least absolute shrinkage and selection operator (LASSO) were used to select radiomics features. Six classifiers were used to train the models (logistic regression (LR), k-nearest neighbor (KNN), decision tree (DT), support vector machine (SVM), random forests (RF), and XGBoost), and then 24 models were established using a random verification method to differentiate low-grade from high-grade meningiomas. The performance was assessed by receiver-operating characteristic (ROC) analysis, the f1-score, sensitivity, and specificity. The radiomics features were significantly associated with the histopathological grading. Quantitative imaging features (n = 1409) were extracted, and nine features were selected to predict the grades of meningiomas. The best performance of the radiomics model for the degree of differentiation was obtained by SVM (area under the curve (AUC), 0.956; 95% confidence interval (CI), 0.83-1.00; sensitivity, 0.87; specificity, 0.92; f1-score, 0.90). The radiomics models are of great value in predicting the histopathological grades of meningiomas, and have broad prospects in radiology and clinics.


Genetic analysis of metabolic cooperation in mammalian cell lines.
Metabolites can be exchanged between cells in culture by direct transfer from the cytoplasm of one cell to that of another in a process known as metabolic cooperation. Most mammalian cell lines are able to transfer small molecules directly between adjacent cells in this way and are consequently mec+; however, a small number are defective in this ability (mec-). Results obtained from somatic cell hybrids formed between combinations of these cells have shown that the four different cell lines examined in this study can be divided into at least two different complementation groups on the basis of their ability to transfer 3H-labeled nucleotides to adjacent cells. Two of the cell lines clearly fall into a single complementation group.


[Suicide attempts in elderly people. Data from the Hospital Italiano de Buenos Aires].
[corrected] The aim of this paper is to analyze some characteristics of 25 patients, over 60, who attempted suicide and were hospitalized in the Department of Psychiatry of a general hospital in Buenos Aires. This is a retrospective study using data from the medical records of patients (1999 to 2002); based on a protocol comprising an analysis of attempted suicides, demographic and clinical variables. Patients were diagnosed following the DSM-IV criteria by two trained GPs and were confirmed by MMPI and Rorschach. 72% were women. The average age was 73,8 % were divorced, 20% never married, 32% widowed and 32% married. The most frequent diagnosis was Major Depressive Disorder of late onset followed by Personality Disorder ( 96%; 48%). The most frequent method was intoxication, (68.75% BDZ). Almost half of the attempts were highly severe. We may infer that the elderly person who attempts suicide: is female, 68-78 years old, does not have a partner, lives with someone, has Major Depressive Disorder (at least half of them had Personality Disorder) and a clinical disease. This is her first attempt, is alone at home, and she does not advise others. Reduced hearing is a very frequent co-morbidity.


Comparison of deep learning segmentation and multigrader-annotated mandibular canals of multicenter CBCT scans.
Deep learning approach has been demonstrated to automatically segment the bilateral mandibular canals from CBCT scans, yet systematic studies of its clinical and technical validation are scarce. To validate the mandibular canal localization accuracy of a deep learning system (DLS) we trained it with 982 CBCT scans and evaluated using 150 scans of five scanners from clinical workflow patients of European and Southeast Asian Institutes, annotated by four radiologists. The interobserver variability was compared to the variability between the DLS and the radiologists. In addition, the generalisation of DLS to CBCT scans from scanners not used in the training data was examined to evaluate its out-of-distribution performance. The DLS had a statistically significant difference (p < 0.001) with lower variability to the radiologists with 0.74 mm than the interobserver variability of 0.77 mm and generalised to new devices with 0.63 mm, 0.67 mm and 0.87 mm (p < 0.001). For the radiologists' consensus segmentation, used as a gold standard, the DLS showed a symmetric mean curve distance of 0.39 mm, which was statistically significantly different (p < 0.001) compared to those of the individual radiologists with values of 0.62 mm, 0.55 mm, 0.47 mm, and 0.42 mm. These results show promise towards integration of DLS into clinical workflow to reduce time-consuming and labour-intensive manual tasks in implantology.


Assessment of child psychological attributes using strength and difficulties questionnaire for prediction of child behavior at first dental visit: a cross-sectional study.
Predicting child behavior before the first dental visit is considered an effective approach that enables the dentist to guide child behavior during the dental treatment. The present study aimed to detect psychological attributes of children seeking dental treatment for the first time using the strength and difficulties questionnaire (SDQ), evaluate actual child behavior during the dental treatment using Frankl Behavior Rating Scale, and test the association between these attributes and child behavior. The study was conducted on 128 children aged between 4 and 7 years attending the outpatient Pediatric Dentistry clinic for the first time. Parents were asked to answer the SDQ, then a simple restorative treatment under local anesthesia was performed for children to evaluate their behavior using Frankl Behavior Rating Scale. About 19.5% of the study sample were categorized as psychologically abnormal, where emotional and conduct problems were the most prevalent psychological attributes by a percentage of 22.7% and 20% respectively. A strong correlation was detected between the total difficulty score and child behavior. Psychological problems are common among children aged from 4 to 7 years and they also affect their dental behavior.


Yellow-colored extract from cashew byproduct - Nonclinical safety assessment.
Natural and synthetic dyes are widely used in foodstuff, medicines and cosmetics industries to enhance and/or restore the color of the final products. This study aimed to evaluate the safety of oral consumption of one carotenoids and anacardic acids-enriched extract (CAE), obtained by green extraction from cashew apple residue fibers, a byproduct of the cashew juice industry. Presenting intense yellow color, CAE could be proposed as a new natural dye. Single and repeated-dose oral toxicity (30 days) were evaluated in female Swiss mice at doses ranging from 50 to 1000 mg/kg, while (anti)mutagenic effects were evaluated in CHO-K1 cells (in vitro Cytokinesis-Block Micronucleus assay - CBMN) and in erythrocytes collected from murine bone marrow (in vivo). CAE did not induce toxic or mutagenic effects in female mice even after 30 days of treatment, regardless of the dose used. Considering cyclophosphamide (CPA)-challenged animals treated with CAE, neither antimutagenic effect was observed nor CAE increased CPA-mutagenic effects although in vitro CBMN results indicated that CAE might increase methyl methanesulfonate-induced micronuclei (MN) frequency besides promoting reduction on CPA-induced MN frequency. The obtained results suggest that CAE may be a safe source of carotenoids with potential use as industrial dye.


Comparing clinical and microbiological methods for the diagnosis of true bacteraemia among patients with multiple blood cultures positive for coagulase-negative staphylococci.
We assessed the accuracy of the Centers for Disease Control and Prevention (CDC) clinical criteria as well as other microbiological methods for the diagnosis of coagulase-negative staphylococci bacteraemia. The CDC clinical criteria had low accuracy, which can be improved by speciation, particularly if the patient had more than two positive blood cultures.


A furanosyl-carbonate autoinducer in cell-to-cell communication of V. harveyi.
An autoinducer arising from reaction of cyclized S-DPD and carbonate is shown to induce light in V. harveyi and thus may play a previously unknown role in quorum sensing.


Iron plaque formed under aerobic conditions efficiently immobilizes arsenic in Lupinus albus L roots.
Arsenic is a non-threshold carcinogenic metalloid. Thus, human exposure should be minimised, e.g. by chemically stabilizing As in soil. Since iron is a potential As immobiliser, it was investigated whether root iron plaque, formed under aerobic conditions, affects As uptake, metabolism and distribution in Lupinus albus plants. White lupin plants were cultivated in a continuously aerated hydroponic culture containing Fe/EDDHA or FeSO4 and exposed to arsenate (5 or 20 μM). Only FeSO4 induced surficial iron plaque in roots. LA-ICP-MS analysis accomplished on root sections corroborated the association of As to this surficial Fe. Additionally, As(V) was the predominant species in FeSO4-treated roots, suggesting less efficient As uptake in the presence of iron plaque. Fe/EDDHA-exposed roots neither showed such surficial FeAs co-localisation nor As(V) accumulation; in contrast As(III) was the predominant species in root tissue. Furthermore, FeSO4-treated plants showed reduced shoot-to-root As ratios, which were >10-fold lower compared to Fe/EDDHA treatment. Our results highlight the role of an iron plaque formed in roots of white lupin under aerobic conditions on As immobilisation. These findings, to our knowledge, have not been addressed before for this plant and have potential implications on soil remediation (phytostabilisation) and food security (minimising As in crops).


Discovery of structurally distinct tricyclic M4 positive allosteric modulator (PAM) chemotypes - Part 2.
This Letter details our efforts to develop novel tricyclic M4 PAM scaffolds with improved pharmacological properties. This endeavor involved a "tie-back" strategy to replace the 3-amino-4,6-dimethylthieno[2,3-b]pyridine-2-carboxamide core which lead to the discovery of two novel tricyclic cores: a 7,9-dimethylpyrido[3',2':4,5]thieno[3,2-d]pyrimidine core and 2,4-dimethylthieno[2,3-b:5,4-c']dipyridine core. Both tricyclic cores displayed low nanomolar potency against the human M4 receptor.


No difference in cognitive performance between phases of the menstrual cycle.
No differences were seen in performance on either verbosequential or visuospatial neuropsychological tests among three groups of women. There were also no differences among the phases of the menstrual cycle on any of the tests. The design of this study was to sample performance of women on verbosequential and visuospatial neuropsychological tests during phases of the normal menstrual cycle in regularly menstruating women, and at comparable time points among women taking oral contraceptives and among those who had amenorrhea. Serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol, and progesterone were assayed to document these hormone levels at each time point for each group. These data are consistent with previous results in our laboratory but conflict with other reports in which shifts in cognitive function were related to fluctuations in estrogen. The only indications of positive results were weak correlations with gonadotropins and cognitive function across subjects. While some of these correlations replicated previous results, they were weak and should be considered with caution.


Measurement of branched-chain ketoacids in plasma by high-performance liquid chromatography.
Branched-chain ketoacids were isolated from plasma or serum samples by acidification, passage through a cationic exchange resin, ether extraction, and extraction of the ether layer with phosphate buffer. The recovery of 2-[1-14C]ketoisocaproate taken through these procedures averaged 95 +/- 3%. Branched-chain ketoacids were measured by high-performance liquid chromatography using a single mobile phase (sodium phosphate:acetonitrile). In normal human subjects, mean +/- SD fasting levels of 2-ketoisocaproate, 2-keto-3-methylvalerate, and 2-ketoisovalerate were 29 +/- 8, 18 +/- 4, and 12 +/- 3 microM, respectively. In normal rats, slightly different results were found: 24 +/- 10, 19 +/- 7 and 17 +/- 6 microM, respectively. In both species, levels of each ketoacid expressed as fractions of total branched-chain ketoacids were much less variable.


Open cavity formed by a photonic crystal with negative effective index of refraction.
An open cavity formed by three 60 degrees wedges of a photonic crystal with negative effective index is designed and studied. The quality factor of the open cavity can be larger than 2000. The influence of the interface termination on the resonant frequency and the quality factor is studied.


COMT haplotypes, catecholamine metabolites in plasma and clinical response in schizophrenic and bipolar patients.
We examined the association of COMT haplotypes and plasma metabolites of catecholamines in relation to the clinical response to antipsychotics in schizophrenic and bipolar patients. We studied 165 patients before and after four weeks of treatment, and 163 healthy controls. We assessed four COMT haplotypes and the plasma concentrations of HVA, DOPAC and MHPG. Bipolar patients: haplotypes are associated with age at onset and clinical evolution. In schizophrenic patients, an haplotype previously associated with increased risk, is related to better response of negative symptoms. Haplotypes would be good indicators of the clinical status and the treatment response in bipolar and schizophrenic patients. Larger studies are required to elucidate the clinical usefulness of these findings.


Effects of inorganic nitrogen sources on the production of PP-V [(10Z)-12-carboxyl-monascorubramine] and the Expression of the nitrate assimilation gene cluster by Penicillium sp. AZ.
A fungal strain, Penicillium sp. AZ, produced the azaphilone Monascus pigment homolog when cultured in a medium composed of soluble starch, ammonium nitrate, yeast extract, and citrate buffer, pH 5.0. One of the typical features of violet pigment PP-V [(10Z)-12-carboxyl-monascorubramine] is that pyranoid oxygen is replaced with nitrogen. In this study, we found that ammonia and nitrate nitrogen are available for PP-V biosynthesis, and that ammonia nitrogen was much more effective than nitrate nitrogen. Further, we isolated nitrate assimilation gene cluster, niaD, niiA, and crnA, and analyzed the expression of these genes. The expression levels of all these genes increased with sodium nitrate addition to the culture medium. The results obtained here strongly suggest that Penicillium sp. AZ produced PP-V using nitrate in the form of ammonium reduced from nitrate through a bioprocess assimilatory reaction.


RNase E levels in Escherichia coli are controlled by a complex regulatory system that involves transcription of the rne gene from three promoters.
The rne gene of Escherichia coli encodes RNase E, an essential endoribonuclease that is involved in both mRNA decay and rRNA processing. Here we present evidence that the gene is transcribed from three promoters: p1, p2 and p3. The p2 and p3 promoters map 34 and 145 nt upstream from the previously characterized rne promoter, p1, generating unusually long 5' UTRs of 395 and 506 nt respectively. Based on promoter-lacZ transcriptional fusions, p1 is a more efficient promoter than either p2 or p3. Low copy number or single copy number vectors carrying rne transcribed from either p1, p2 or p3 alone complement the rne 1018::bla deletion mutation at 30 degrees C, 37 degrees C and 44 degrees C. However, normal autoregulation requires the presence of all three promoters. A comparison among intracellular levels of RNase E, the half-lives of the rpsO, rpsT and rne mRNAs, and growth rates, indicates that the cell contains a considerable excess of RNase E protein. In addition, when the rne transcript is stabilized at low RNase E levels, it is not efficiently translated.


Role of imaging in the management of pancreatic mass.
Pancreatic cancer is the second commonest malignant gastrointestinal neoplasm. Modern imaging techniques have greatly increased sensitivity in diagnosing and staging pancreatic cancers. Multidetector CT in particular, plays a critical role in local staging and determining the resectability of pancreatic tumours. MR and endoscopic ultrasound are valuable in those groups of patients in whom CT findings alone are inconclusive in tumour characterisation and local staging, particularly vascular involvement. In this article we review the current established concepts and the role of imaging in the multidisciplinary management of pancreatic tumours together with a comprehensive review of the literature.


Carbon dioxide uroflowmetry.
A method using pneumatic compression to determine flow rate is described. The procedure is best suited for physicians using carbon dioxide instrumentation for a urodynamic assessment of patients since the carbon dioxide cystometer is readily converted to a uroflowmeter. Values obtained with this method are similar to values reported in the literature using other techniques. When voided volumes are more than 200 ml. the values for peak flow rate are reproducible within subjects and are volume-dependent.


The management of leaking rectal anastomoses by minimally invasive techniques.
The salvage of a leaking rectal anastomosis has been reported in selected cases. Herein we present our experience in the management of rectal anastomotic dehiscence by minimally invasive techniques. A total of 94 patients underwent anterior rectal resection for rectal cancer in an 18-month period. The operation was laparoscopic-assisted in 76 cases (81%). Clinical anastomotic dehiscences occurred in 10 cases (10.6%). Conservative therapy, including the percutaneous drainage of pelvic abscesses, was successful in three cases. Among the seven patients who required reoperation, the anastomosis was taken down in only one case. In four of these cases, the reoperation was performed laparoscopically, including peritoneal lavage and the creation of a diverting stoma. There were no complications resulting from the use of laparoscopy in the early postoperative period. Closure of the stoma was possible in eight patients. The salvage of a leaking rectal anastomosis is feasible in the vast majority of these cases (80%). The combination of laparoscopy and interventional radiology is a good alternative to laparotomy in patients who have had previous laparoscopic rectal excision.


Nocardial osteomyelitis: a case report and review of the literature.
The occurrence of human nocardiosis is increasing in both immunocompetent as well as immunosuppressed patients. A 63-yr-old male construction worker presented with complaints of left hip pain and rapidly enlarging masses on his left neck and chest. He had been treated for a pneumonia of unknown etiology 3 mo prior to admission. A debridement of the left femur was performed and the curetted material was positive for Nocardia asteroides. Osteomyelitis due to the hematogenous spread of Nocardia asteroides is rare, but will undoubtedly be seen with increasing frequency. The literature, as well as 11 reported cases of the hematogenous spread of this organism, are reviewed.