Benzodiazepine prescribing in elderly Australian general practice patients.
The use of benzodiazepines by elderly people is of limited therapeutic benefit and increases the risk of adverse events. This study aimed to examine the extent to which benzodiazepines are prescribed for elderly Australians. Data for 3,970 individuals aged 65 years or more were extracted from a general practice database. Benzodiazepine prescriptions for 2002 were reviewed. Overall, 16% (95% CI 11-21%) of elderly patients had at least one benzodiazepine prescription. Females were almost twice as likely as males to be prescribed a benzodiazepine and prescription prevalence increased with age. Despite risks, benzodiazepines are widely prescribed for the elderly. Limited availability and cost of alternative therapies and pressures on the primary care system in Australia may contribute to their continued overuse. The prescribing of benzodiazepines for elderly Australians needs to be reduced by better managing sleep and anxiety problems.


Blood pressure variability and risk of dementia in an elderly cohort, the Three-City Study.
The relationship between blood pressure and dementia is incompletely understood in elderly individuals. Blood pressure variability may have a role in the risk of dementia. This investigation was a cohort study of 6506 elderly individuals followed-up for 8 years (1999-2001 through 2008) with assessments at years 2, 4, and 7-8. Blood pressure was measured by electronic devices at baseline and at 2- and 4-year follow-up examinations. Cox proportional hazard models adjusted for potential confounders were used to estimate the risk of incident dementia according to blood pressure (means and coefficients of variation of the three measures). During the 40,151 person-years of follow-up 474 participants developed dementia. We observed no association between mean blood pressure and risk of dementia. In contrast, an increase of 1 standard deviation in the coefficient of variation of blood pressure was associated with a 10% increased risk of dementia. Analysis by deciles of the coefficient of variation showed that the higher the variability, the higher the risk of dementia (P<.02 for trend). In the fully adjusted Cox model, the risk of dementia for those in the highest decile of the coefficient of variation of systolic blood pressure was 1.77 (1.17-2.69) compared with the lowest decile. In this cohort study, variability of blood pressure during follow-up was associated with an increased risk of incident dementia, whereas mean blood pressure was not. Limitation of blood pressure fluctuation may be an important target to preserve cognitive function in the elderly.


A hotspot for novel amplification joints in a mosaic of Alu-like repeats and palindromic A + T-rich DNA.
We have identified, in the amplified domain of adenylate deaminase (AMPD) overproducing Chinese hamster fibroblasts, a 2.6 kb recombinogenic DNA region which is frequently involved in amplification-associated rearrangements. The nucleotide sequence reveals a mosaic organization of four Alu-equivalent repeats of the B1 and B2 families and eight long A + T-rich DNA segments. Part of this region is enriched with long imperfect palindromes. The center of one palindrome contains a putative topoisomerase I cleavage site and this site defines the position of a novel junction which was formed by illegitimate recombination with anther A + T-rich DNA sequence located far apart on the amplified DNA. These findings and their significance are discussed in the context of related data from other systems and in the light of current models for eukaryotic DNA recombination, replication and organization.


Fast immobilized liposome chromatography based on penetrable silica microspheres for screening and analysis of permeable compounds.
In the present study, an immobilized liposome chromatography (ILC) stationary phase based on penetrable silica microspheres was prepared. The silica possessed mesopores and penetrable macropores, which afforded sufficient surface area and fast mass transfer, respectively. Compared with the ILC column based on common porous silica gels, the penetrable silica had larger capacity for liposomes' immobilization, lower backpressure and higher separation efficiency. Twenty-two kinds of drugs were used as tested analytes and their retention behaviors on newly prepared ILC column were investigated in detail. Column temperature and pH of mobile phase were both key factors affecting the retention of solutes. The retention of these drugs on ILC column reflected their permeability in vivo. Furthermore, the methanolic aqueous extracts of three traditional Chinese medicines (TCMs), Radix Liquiritiae, Scutellaria Baicalensis and Flos Sophorae, were screened on this novel ILC column. Effects of column temperature and eluent pH on chromatographic behaviors of components of the TCM methanolic extracts were also studied. Several permeable components can be found, indicating that they are potentially active components in the TCMs. It is also found that the as-prepared ILC column remained stable in at least 1 month and the relative standard deviations of adjusted retention times for solutes were lower than 9.0%. The proposed ILC based on the penetrable silica microspheres is promising for the fast and low-pressure separation, especially for fast screening of permeable compounds and modeling the drug-membrane interaction in vitro. It would be a useful approach to predict the permeability of potential active drugs.


Efficacy of initial caspofungin plus trimethoprim/sulfamethoxazole for severe PCP in patients without human immunodeficiency virus infection.
The number of pneumocystis pneumonia (PCP) cases is increasing in immunocompromised patients without human immunodeficiency virus infection (HIV), causing serious morbidity with high mortality. Trimethoprim/sulfamethoxazole (TMP/SMZ) monotherapy has limited effectiveness in the treatment of PCP. Clinical data on whether initial caspofungin plus TMP/SMZ for this disease is superior to monotherapy in non-HIV-infected patients are limited. We aimed to compare the clinical effectiveness of these regimens for severe PCP in non-HIV patients. A retrospective study reviewed 104 non-HIV-infected patients with confirmed PCP in the intensive care unit between January 2016 and December 2021. Eleven patients were excluded from the study because TMP/SMZ could not be used due to severe hematologic disorders or clinical data were missing. All enrolled patients were divided into three groups according to different treatment strategies: Group 1 received TMP/SMZ monotherapy, Group 2 received caspofungin combined with TMP/SMZ as first-line therapy, and Group 3 initially received TMP/SMZ monotherapy and later received caspofungin as salvage therapy. The clinical characteristics and outcomes were compared among the groups. A total of 93 patients met the criteria. The overall positive response rate of anti-PCP treatment was 58.06%, and the overall 90-day all-cause mortality rate was 49.46%. The median APACHE II score was 21.44. The concurrent infection rate was 74.19%, among whom 15.05% (n = 14) of those patients had pulmonary aspergillosis, 21.05% (n = 20) had bacteremia, and 23.65% (n = 22) had CMV infections. The patients who received initial caspofungin combination with TMP/SMZ had the best positive response rate (76.74%) compared to others (p = 0.001). Furthermore, the group that received initial caspofungin combined with TMP/SMZ had a 90-day all-cause mortality rate (39.53%) that was significantly different from that of the shift group (65.51%, p = 0.024), but this rate showed no statistically significant difference compared with that in the monotherapy group (48.62%, p = 0.322). None of the patients had serious adverse events from caspofungin therapy. For non-HIV-infected patients with severe PCP, initial combination therapy with caspofungin and TMP/SMZ is a promising first-line treatment option compared with TMP/SMZ monotherapy and combination therapy as salvage therapy.


Differences in neuronal differentiation between the transient cranial (Frorieps') and normal dorsal root ganglia.
The Frorieps' ganglia are dorsal root ganglia (DRG) that form, grow slowly compared to normal DRG, and then degenerate during normal embryonic development of amniotes. Their fate has been shown to be regulated by the Hox family and other genes involved in pattern formation, and by the mesodermal microenvironment of the cranial somites in which they develop. Neurons are known to differentiate within the Frorieps' DRG in the course of their short life span. In this study, we compared several aspects of neural differentiation between the longest-lived Frorieps' ganglia, DRG2, and normal trunk DRG in chick embryos. The expression of transcription factor Islet-1 and the RNA-binding protein Hu differ between normal and Frorieps' DRG at St. 23 (embryonic day 4). Islet-1 is expressed in a higher proportion of cells in DRG2 than DRG5, suggesting that cells in DRG2 differentiate more rapidly into neurons than in normal DRG. This molecular difference appears at the same developmental stage as overt morphological differences between DRG2 and normal DRG. Other LIM-homeobox proteins (Lim-1, -2 and -3 and Islet-2) are not expressed either in normal or Frorieps' DRG at early stages of development. Somite-grafting experiments reveal that the increased proportion of Isl-1(+) in DRG2 is due to the special microenvironment of the cranial somites. In contrast to Islet-1, the Hu antigen is expressed in a slightly lower proportion of cells in DRG2 at St. 23. At St. 28, there is a significant population of neurons in DRG2 that are Isl-1(+)/Hu(-). Since Islet-1 is normally expressed before Hu in DRG cells, it is possible that the reduced growth rate of the DRG2 ganglion may partially result from the inability of precociously differentiating Islet-1(+) neurons to further mature, as reflected by Hu antigen expression. In contrast to the neuronal markers examined, the satellite cell marker 7B3 is expressed at similar levels in DRG2 and DRG5.


Characterization of immunoglobulin class and subclass responses in secondary aborter sera.
Sera from secondary (2 degrees) aborters exhibit persistent, high-titred cytotoxicity against paternal as well as HLA dissimilar non-paternal lymphocytes. The majority of antipaternal complement-dependent cytotoxicity (CDC) and complement-independent antibody dependent cellular cytotoxicity (ADCC) was recovered in IgG enriched fractions following ion-exchange chromatography of 2 degrees aborter sera. The IgG subclasses mediating antipaternal reactivity were determined using murine anti-human IgG subclass specific monoclonal antibodies and Protein A affinity chromatography (SPA). Inhibition of paternal CDC by the anti-subclass reagents showed 75-90% of the reactivity mediated by maternal IgG1 antibodies. Anti-IgG3 inhibited 15-30% whereas anti-IgG2 produced little inhibition. SPA chromatography of 2 degrees aborter IgG supported the monoclonal antibody results in that greater than 80% of the CDC activity was recovered in the IgG1, 2, and 4 containing eluate and 20% was found in the IgG3 enriched effluent. Although the anti-IgG subclass specific monoclonals did not inhibit antipaternal ADCC, IgG3 did not appear to mediate this cytotoxicity as the ADCC was recovered in the eluate and not the effluent following SPA chromatography of 2 degrees aborter IgG enriched serum fractions. These data indicate that the humoral antipaternal and polyspecific CDC immune reactivities of 2 degrees aborters are due to the production of IgG1 and IgG3 antibodies.


Human neutrophils degrade methacrylate resin composites and tooth dentin.
Cholesterol esterase-like (CE) activity from saliva and esterase from cariogenic bacteria hydrolyze ester linkages of dental methacrylate resins. Collagenolytic, matrix metalloproteinase-like (MMP) activities from dentin and bacteria degrade collagen in demineralized tooth dentin. Human neutrophils in the oral cavity contain factors that are hypothesized to have CE and MMP activities that could contribute to the degradation of methacrylate resins and dentinal collagen. OBJECTIVES: To measure the CE and MMP activities from human neutrophils and their ability to degrade dental methacrylate resin composite and dentinal collagen. Neutrophils' CE and MMP activities were measured using nitrophenyl-esters or fluorimetric MMP substrates, respectively. Neutrophils' degradation of resin composite and dentinal collagen was quantified by measuring release of a universal 2,2-Bis[4-(2-hydroxy-3-methacryloxypropoxy)phenyl]propane (bisGMA)-derived resin composite degradation byproduct, bishydroxy-propoxy-phenyl-propane (bisHPPP), or a collagen degradation by-product, hydroxyproline, respectively using ultra performance liquid chromatography/mass spectrometry. Neutrophils' CE activity increased the release of bisHPPP from bisGMA monomer compared to control after 24 and 48 h (p < 0.05). Neutrophils degraded polymerized resin composite and produced higher amounts of bisHPPP than buffer after 48 h of incubation (p < 0.05). Neutrophils show generic MMP, gelatinase, MMP-2 and MMP-9, and collagenase, MMP-1 and MMP-8 activities that were stable or increased over the first 24 h (p < 0.05). Neutrophils degraded demineralized dentin more than buffer-only groups, indicated by higher amounts of hydroxyproline (p < 0.05). The ability of neutrophils to degrade both dental resin composite and tooth dentin, suggest neutrophil's potential role in root caries, and in recurrent carries by accelerating the degradation of resin-dentin interfaces, and compromising the longevity of the restoration. STATEMENT OF SIGNIFICANCE: Neutrophils are part of the innate immune system and are constantly entering the oral cavity through the gingival sulcus, in direct contact with the tooth, restoration, restoration-tooth margins and pathogenic bacteria. The current study is the first to characterize and quantify degradative activities from neutrophils toward methacrylate resin and demineralized dentin, the two main components of the restoration-tooth interface, suggesting that this interface could be negatively influenced by neutrophils, potentially contributing to increase in caries formation and progression, and premature restoration failure. This study provides a significant finding to the biomaterials and oral health fields by identifying a potential weakness in current restorative procedures and materials used to manage gingival proximal and cervical gingival or sub-gingival carious lesions.


Factors associated with HIV testing among public sector clinic attendees in Johannesburg, South Africa.
Uptake of VCT remains low in many sub-Saharan African countries. Men and women aged 15 and older were recruited from a family planning, STI, and VCT clinic in inner-city Johannesburg between 2004 and 2005 to take part in a cross-sectional survey on HIV testing (n = 198). Fourty-eight percent of participants reported previously testing for HIV and, of these, 86.9% reported disclosing their status to their sex partner. In multivariable analyses, individuals whose partners had been tested for HIV were more likely to have tested (AOR 2.92; 95% CI: 1.38-6.20). In addition, those who reported greater blame/shame attitudes towards people living with HIV/AIDS were less likely to have tested (AOR 0.35; 95% CI: 0.16-0.77) while those reporting more equitable attitudes towards people living with HIV/AIDS were more likely to have tested (AOR 2.87; 95% CI: 1.20-6.86). Promotion of and increased access to couples HIV testing should be made available within the South African context.


Binding of (3H)dihydroalprenolol to beta adrenoceptors of cells isolated from adult rat heart.
Myocardial cells isolated from adult rat heart bind (3H)dihydroalprenolol. Sixty-one percent of this binding appeared to be at the beta adrenoceptors since it was inhibited by saturating quantities of the beta antagonist propranolol or by the beta agonist isoprenaline. The binding is stereoselective as the l-isomer of isoprenaline caused greater inhibition than the d-isomer. The binding of (3H)dihydroalprenolol to beta adrenoceptors was saturable; half maximum binding occurred at about 8 nM and full saturation at 30--40nM.


Co-localization of proteins with defined sequential order and controlled stoichiometric ratio on magnetic nanoparticles.
Co-immobilization of enzymes used in cascade reactions is important for improving the overall catalytic efficiency. In this work, we employed scaffoldins as a bridge and succeeded in a highly-ordered co-localization of multiple proteins on magnetic nanoparticles with a loading capacity of ∼0.831 μmol g-1 supports.


Comparison of the Kato-Katz thick smear and McMaster egg counting techniques for monitoring drug efficacy against soil-transmitted helminths in schoolchildren on Pemba Island, Tanzania.
The Kato-Katz thick smear technique is widely used to assess prevalence and intensity in soil-transmitted helminth (STH) control programmes, but its usefulness in monitoring anthelminthic drug efficacy needs to be validated and compared with other methods. A promising alternative is the McMaster egg counting technique. In the present study, the efficacy of single-dose albendazole against STH infections in 430 schoolchildren on Pemba Island was assessed using both the Kato-Katz and McMaster techniques. The study revealed comparable drug efficacy results for both methods and confirmed the potency of the McMaster technique as an alternative method for monitoring large-scale deworming programmes.


Loss of virulence in Shigella strains preserved in culture collections due to molecular alteration of the invasion plasmid.
Fifty-two Shigella strains long preserved by three Japanese culture collections were examined for virulence. All of them were avirulent when judged by the focus-plaque assay and the ability to bind Congo red. Fifteen strains had a plasmid comparable in size to that responsible for epithelial invasiveness and were positive in hybridization tests with a probe derived from a plasmid cistron, virG. Twenty-four strains had a similar plasmid but were negative in hybridization tests. The remaining 13 strains were negative in all the five criteria for virulence. Similar studies made on one hundred Shigella strains isolated from 1967 to 1985 clearly demonstrated loss of virulence with prolonged time of storage.


Lipoprotein receptors in oocyte growth.
The transport and removal of lipoproteins from the blood in the laying hen is of particular interest because it is a system in which the massive transport of lipid to one organ (the ovary) coexists with regulatory mechanisms for the control of lipid homeostasis in extraovarian tissues. In order to achieve this dual task, the laying hen expresses dichotomous receptor-mediated pathways. On one hand, very low density lipoproteins (VLDL) and vitellogenin (VTG), which together form over 95% of the lipid in a fully grown oocyte (i.e., an egg yolk), are transported into oocytes via a 95-kDa receptor protein. This receptor, termed oocyte VLDL/VTG receptor, is exclusively produced in growing oocytes and is absent from somatic cells. It shows a high degree of structural similarity to other members of the so-called low density lipoprotein (LDL) receptor family, but in contrast to the LDL receptor, its expression is not suppressed by sterols. On the other hand, somatic cells, but not oocytes, synthesize a 130-kDa receptor that recognizes VLDL-derived, cholesterol-rich lipoproteins. This receptor is the functional analogue to the mammalian LDL receptor in that it mediates, at least in part, extraoocytic lipid homeostasis. The somatic LDL receptor of the chicken recognizes apolipoprotein (apo)B, but not VTG, in accordance with VTG's exclusive routing to growing oocytes. Within oocytes, both apoB of VLDL and VTG undergo limited specific postendocytic proteolytic processing. Recent studies have shown that this breakdown of macromolecular plasma precursor molecules is catalyzed by an endosomal form of cathepsin D and is a key event in the formation of yolk, the major nutrient source for the developing embryo.


Recognizing large-cell transformation of mycosis fungoides.
Although patients with mycosis fungoides (MF) typically experience an indolent disease course, a minority undergo a process of large-cell transformation (LCT), which often heralds more aggressive disease and shortened survival. Regrettably, most dermatologists are unfamiliar with LCT, and even fewer understand how to recognize it clinically. Because a diagnosis of LCT typically triggers more aggressive therapy and/or referral to cutaneous T-cell lymphoma (CTCL) centers, it is paramount for clinicians to be able to recognize suspect lesions visually. LCT is diagnosed histologically; however, diagnostic biopsy is performed only if transformed lesions are suspected clinically. Because the literature provides little information on what clinical features should lead to suspicion of LCT, we sought to identify and categorize the presentations of LCT to aid in its recognition. We identified 14 patients with biopsy-proven LCT confirmed by a board-certified dermatopathologist experienced with this diagnosis. The clinical presentations of LCT, timing of its evolution, and treatment regimens were evaluated by chart and photograph review. We devised 3 categories that clinically represent LCT: (1) LCT occurring as a new, solitary nodule within a classic MF patch or plaque, (2) LCT occurring as abrupt onset of multiple scattered papules and/or nodules without spontaneous resolution, and (3) LCT occurring within new or enlarging tumors. A larger number of reviewed cases might reveal additional clinical presentations of LCT. Dermatologists may use our categories of clinical indicators to recognize and diagnose LCT earlier, allowing implementation of more aggressive treatment regimens when appropriate.


The state-of-the-science: challenges in designing postacute care payment policy.
This report describes Medicare's postacute care (PAC) payment systems and their incentives, as well as global changes in capacity, quality, and utilization over time. It assesses the payment systems' impact on PAC services, referencing relevant works in progress. Suggestions are made for future research.


Sleep Fragmentation and Estradiol Suppression Decrease Fat Oxidation in Premenopausal Women.
Body fat gain associated with menopause has been attributed to estradiol (E2) withdrawal. Hypoestrogenism is unlikely to be the only contributing factor, however. Given the links between sleep and metabolic health, we examined the effects of an experimental menopausal model of sleep fragmentation on energy metabolism. Twenty premenopausal women (age 21-45 years) underwent a 5-night inpatient study during the mid-to-late follicular phase (estrogenized; n = 20) and the same protocol was repeated in a subset of the participants (n = 9) following leuprolide-induced E2 suppression (hypo-estrogenized). During each 5-night study, there were 2 nights of unfragmented sleep followed by 3 nights of fragmented sleep. Indirect calorimetry was used to assess fasted resting energy expenditure (REE) and substrate oxidation. Sleep fragmentation in the estrogenized state increased the respiratory exchange ratio (RER) and carbohydrate oxidation while decreasing fat oxidation (all P < 0.01). Similarly, in the hypo-estrogenized state without sleep fragmentation, RER and carbohydrate oxidation increased and fat oxidation decreased (all P < 0.01); addition of sleep fragmentation to the hypo-estrogenized state did not produce further effects beyond that observed for either intervention alone (P < 0.05). There were no effects of either sleep fragmentation or E2 state on REE. Sleep fragmentation and hypoestrogenism each independently alter fasting substrate oxidation in a manner that may contribute to body fat gain. These findings are important for understanding mechanisms underlying propensity to body fat gain in women across the menopause transition.


Validation of a liquid chromatography/tandem mass spectrometry method for the simultaneous quantification of Sotrastaurin and its metabolite N-desmethyl-sotrastaurin in human blood.
A liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) method was validated for the quantification of Sotrastaurin (AEB071) and N-desmethyl-sotrastaurin in human blood. The validation of the analytical procedure was performed according to the latest Food and Drug Administration (FDA) "Guidance for Industry, Bioanalytical Method Validation". Chromatographic separation was performed using an RP C₁₈ (50 mm × 4.6 mm, 5 μm) column at 40±3.0 °C with a mobile phase consisted of 2 mM ammonium acetate in water (pH 4.5):methanol:acetonitrile (25:15:60, v/v) of a flow rate of 1 mL/min followed by quantification with tandem mass spectrometer, operated in electrospray ionization (ESI) positive ion mode and applying multiple reaction monitoring (MRM). The validated method described in this paper presents high absolute recovery, with a sensitivity of 3.00 ng/mL as lower limit of quantitation using a sample volume of 300 μL, low inter-run bias and variability (for Sotrastaurin, -4.4 to 0.4% and 1.8 to 2.5% and for N-desmethyl-sotrastaurin, ranged from 1.6 to 2.3% and 2.7 to 3.9%, respectively) with a short runtime of 3.5 min. The method was validated using K₃EDTA as specific anticoagulant and cross-validated using Li-Heparin and Na-Heparin. The method was specific for Sotrastaurin and N-desmethyl-sotrastaurin within the given criteria of acceptance (apparent peak area for Sotrastaurin and N-desmethyl-sotrastaurin in zero samples ≤ 20% of mean peak area at LLOQ) in human blood. The method was fully validated for the quantitative determination of Sotrastaurin and its metabolite N-desmethyl-sotrastaurin in human blood between the range of 3.00 ng/mL and 1200 ng/mL.


Self-monitoring during spelling practice: effects on spelling accuracy and on-task behavior of three students diagnosed with attention deficit hyperactivity disorder.
We used a multiple baseline across subjects design to investigate the effects of self-monitoring on the on-task behavior and spelling accuracy of three fifth-grade students diagnosed with attention deficit hyperactivity disorder (ADHD) during an independent spelling practice period in the general education environment. We also compared their on-task levels to that of peers without disabilities in the same classroom context before and after the self-monitoring intervention. Our results showed that time on-task and spelling accuracy increased for the students diagnosed with ADHD during independent spelling practice after the teacher taught them how to self-monitor. These same students' on-task behavior also increased to levels comparable to that of their peers. Suggestions for educators interested in teaching their students self-monitoring strategies are included following a discussion of the results of the analysis.


Beighton Score, Tibial Slope, Tibial Subluxation, Quadriceps Circumference Difference, and Family History Are Risk Factors for Anterior Cruciate Ligament Graft Failure: A Retrospective Comparison of Primary and Revision Anterior Cruciate Ligament Reconstructions.
To assess patient history, physical examination findings, magnetic resonance imaging (MRI) and 3-dimensional computed tomographic (3D CT) measurements of those with anterior cruciate ligament (ACL) graft failure compared with primary ACL tear patients to better discern risk factors for ACL graft failure. We performed a retrospective review comparing patients who underwent revision ACL reconstruction (ACLR) with a primary ACLR group with minimum 1-year follow-up. Preoperative history, examination, and imaging data were collected and compared. Measurements were made on MRI, plain radiographs, and 3D CT. Inclusion criteria were patients who underwent primary ACLR by a single surgeon at a single center with minimum 1-year follow-up or ACL graft failure with revision ACLR performed by the same surgeon. A total of 109 primary ACLR patients, mean age 33.7 years (range 15 to 71), enrolled between July 2016 and July 2018 and 90 revision ACLR patients, mean age 32.9 years (range 16 to 65), were included. The revision ACLR group had increased Beighton score (4 versus 0; P < .001) and greater side-to-side differences in quadricep circumference (2 versus 0 cm; P < .001) compared with the primary ACLR group. A family history of ACL tear was significantly more likely in the revision group (47.8% versus 16.5%; P < .001). The revision group exhibited significantly increased lateral posterior tibial slope (7.9° versus 6.2°), anterolateral tibial subluxation (7.1 versus 4.9 mm), and anteromedial tibia subluxation (2.7 versus 0.5 mm; all P < .005). In the revision group, femoral tunnel malposition occurred in 66.7% in the deep-shallow position and 33.3% in the high-low position. The rate of tibial tunnel malposition was 9.7% from medial to lateral and 54.2% from anterior to posterior. Fifty-six patients (77.8%) had tunnel malposition in ≥2 positions. Allograft tissue was used for the index ACLR in 28% in the revision group compared with 14.7% in the primary group. Beighton score, quadriceps circumference side-to-side difference, family history of ACL tear, lateral posterior tibial slope, anterolateral tibial subluxation, and anteromedial tibia subluxation were all significantly different between primary and revision ACLR groups. In addition, there was a high rate of tunnel malposition in the revision ACLR group.


Survival after resection of stage II non-small cell lung cancer.
From 1973 to 1989, 214 patients with stage II non-small cell lung cancer were treated by resection and complete mediastinal lymph node dissection. There were 116 adenocarcinomas and 98 squamous cancers. There were 35 T1 N1 and 179 T2 N1 tumors. Whereas T1 tumors were mainly adenocarcinomas (83%), this difference was not apparent in T2 lesions. Regardless of histology, half of the patients had a single involved N1 lymph node. Lobectomy was performed in 68% of the patients, pneumonectomy in 31%, and wedge resection or segmentectomy in 1%. Lobectomy was sufficient to encompass all disease in 34 of 35 T1 N1 tumors. Only 48 patients (22%) received postoperative external irradiation and 11 patients (5%) received chemotherapy. The overall 5-year disease-free survival was 39%. The best survival rates were in patients who had a single node involved and tumors 3 cm or less in diameter (48%). The pattern of recurrence differed by histology. Local or regional recurrence was more frequent in patients with squamous carcinoma whereas distant metastases were more commonly seen in adenocarcinomas (87%) with brain as the most frequent site (adenocarcinoma, 52%; squamous, 34%). It is concluded that in stage II carcinomas, resection remains the treatment of choice, that mediastinal lymph node dissection provides the most accurate staging, and that the best adjuvant treatment to improve survival is yet to be determined.


Copper handling by astrocytes: insights into neurodegenerative diseases.
Copper (Cu) is an essential trace element in the brain that can be toxic at elevated levels. Cu accumulation is a suspected etiology in several neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, and prion-induced disorders. Astrocytes are a proposed depot in the brain for Cu and other metals, including lead (Pb). This article describes the physiological roles of Cu in the central nervous system and in selected neurodegenerative diseases, and reviews evidence that astrocytes accumulate Cu and protect neurons from Cu toxicity. Findings from murine genetic models of Menkes disease and from cell culture models concerning the molecular mechanisms by which astrocytes take up, store, and buffer Cu intracellularly are discussed, as well as potential mechanistic linkages between astrocyte functions in Cu handling and neurodegenerative diseases.


Magnetic graphitized carbon black based on crystal growth method combined with high-resolution mass spectrometry for screening of 300 pesticide residues in Radix Codonopsis and Angelica sinensis.
In this study, a high-throughput method for analyzing 300 pesticide residues in Radix Codonopsis and Angelica sinensis was established by liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS) using iron tetroxide loaded graphitized carbon black magnetic nanomaterial (GCB/Fe3O4) as the purification material. It was optimized that saturated salt water and 1 % acetate acetonitrile were used as the extraction solution, then the supernatant was purified with 2 g anhydrous CaCl2 and 300 mg GCB/Fe3O4. As a result, 300 pesticides in Radix Codonopsis and 260 in Angelica sinensis achieved satisfactory results. The limits of quantification of 91 % and 84 % of the pesticides in Radix Codonopsis and Angelica sinensis reached 10 μg/kg, respectively. The matrix-matched standard curves ranging from 10 to 200 μg/kg were established with correlation coefficients (R) above 0.99. The pesticides meeting SANTE/12682/2021 accounted for 91.3 %, 98.3 %, 100.0 % and 83.8 %, 97.3, 100.0 % of the total pesticides added in Radix Codonopsis and Angelica sinensis respectively, which were spiked at 10, 20,100 μg/kg. The technique was applied to screen 20 batches of Radix Codonopsis and Angelica sinensis. Five pesticides were detected, three of which were prohibited according to the Chinese Pharmacopoeia (2020 Edition). The experimental results showed that GCB/Fe3O4 coupled with anhydrous CaCl2 exhibited good adsorption performance and could be used for sample pretreatment of various pesticide residues in Radix Codonopsis and Angelica sinensis. Compared with the reported methods for determining pesticides in traditional Chinese medicine (TCM), the proposed method has the advantage of less time-consuming in the clean-up procedure. Furthermore, as a case study on root TCM, this approach may serve as a reference for other TCM.


Omental cakes in American Burkitt lymphoma. Computed tomography demonstration.
Diffuse tumor infiltration of the omentum or so-called omental "cakes" is well recognized on computed tomography (CT) and is usually associated with pelvic or gastrointestinal carcinoma. However, this has not been described in American Burkitt lymphoma despite frequent intraabdominal involvement in this disease. We report two cases of American Burkitt lymphoma in which omental cakes were demonstrated by CT.


Acute traumatic intraventricular hemorrhage in children.
Four cases are reported of acute traumatic intraventricular hemorrhage in children revealed by computed tomography (CT). In most cases, the intraventricular hemorrhage had been produced by severe sagittal impact. All of the children were 7 or less on the Glasgow Coma Scale (GCS) on admission. Three had hemorrhage in all ventricles, and one had an intraventricular hemorrhage in the lateral and fourth ventricles. One of them showed enhancement of the blood-filled ventricles, with the appearance of increased ventricular size, on the contrast enhancement CT. Two children with ventricular dilation died soon after injury, although ventricular drainage had been performed; one child remained in a persistent vegetative state, and the other enjoyed good recovery with conservative therapy. Certain features of acute traumatic intraventricular hemorrhage in children are discussed in relation to this series.


Rapid activation of ornithine decarboxylase by mitogenic (but not by nonmitogenic) ligands in human T lymphocytes.
The T cell mitogens, concanavalin A and the monoclonal antibody OKT3, cause a rapid activation of ornithine decarboxylase (ODC) activity in human T lymphocytes, maximal within 10 min of mitogen addition. Nonmitogenic ligands to T cell surface structures do not induce ODC. The enzyme induction is dependent upon an intact mobility of ligand-receptor complex, requires a functioning energy metabolism, but is independent of de novo protein synthesis. The early induction of pre-existing ODC molecules appears to be specifically linked to the initiation of T lymphocyte proliferation.


Progressive loss of corneal nerve fibers is associated with physical inactivity and glucose lowering medication associated with weight gain in type 2 diabetes.
Limited studies have identified risk factors linked to the progression of diabetic peripheral neuropathy (DPN) in type 2 diabetes. This study examined the association of risk factors with change in neuropathy measures over 2 years. Participants with type 2 diabetes (n = 78) and controls (n = 26) underwent assessment of clinical and metabolic parameters and neuropathy using corneal confocal microscopy (CCM), vibration perception threshold (VPT), and the DN4 questionnaire at baseline and 2 year follow-up. Participants with type 2 diabetes had a lower corneal nerve fiber density (CNFD), branch density (CNBD), and fiber length (CNFL) (P ≤ 0.0001) and a higher VPT (P ≤ 0.01) compared with controls. Over 2 years, despite a modest reduction in HbA1c (P ≤ 0.001), body weight (P ≤ 0.05), and LDL (P ≤ 0.05) the prevalence of DPN (P = 0.28) and painful DPN (P = 0.21) did not change, but there was a significant further reduction in CNBD (P ≤ 0.0001) and CNFL (P ≤ 0.05). CNFD, CNBD, and CNFL decreased significantly in physically inactive subjects (P < 0.05-0.0001), whilst there was no change in CNFD (P = 0.07) or CNFL (P = 0.85) in physically active subjects. Furthermore, there was no change in CNFD (P = 0.82), CNBD (P = 0.08), or CNFL (P = 0.66) in patients treated with glucose lowering medication associated with weight loss, whilst CNBD (P = 0.001) decreased in patients on glucose lowering medication associated with weight gain. In participants with type 2 diabetes, despite a modest improvement in HbA1c, body weight, and LDL there was a progressive loss of corneal nerve fibers; except in those who were physically active or on glucose lowering medication associated with weight loss.


Electrospray deposition as a smart technique for laccase immobilisation on carbon black-nanomodified screen-printed electrodes.
Enzymes immobilisation represents a critical issue in the design of biosensors to achieve standardization as well as suitable analytical performances in terms of sensitivity, selectivity, and stability. In this work electrospray deposition (ESD) has been exploited as a novel technique for the immobilisation of laccase enzyme on carbon black modified screen-printed electrodes. The aim is to fabricate an amperometric biosensor for phenolic compound detection. The electrodes produced by ESD have been analysed by scanning electron microscopy and characterised electrochemically to prove that this immobilisation technique is suited to manufacture high performance biosensors. The results show that the laccase enzyme maintains its activity after undergoing the electrospray ionisation process and deposition and the fabricated biosensor has improved performances in terms of storage (up to 3 months at room temperature) and working (up to 25 measurements on the same electrode) stability. The laccase-based biosensor has been tested for phenolic compound detection, with catechol as target analyte, in the linear range 2.5-50 μM, with 2.0 μM limit of detection, without interference from lead, cadmium, atrazine, and paraoxon, and without matrix effect in drinking, surface, and wastewater.


Investigating the Significance of Aerosols in Determining the Coronavirus Fatality Rate Among Three European Countries.
The coronavirus pandemic has not only gripped the scientific community in the search for a vaccine or a cure but also in attempts using statistics and association analysis-to identify environmental factors that increase its potency. A study by Ogen (Sci Total Environ 726:138605, 2020a) explored the possible correlation between coronavirus fatality and high nitrogen dioxide exposure in four European countries-France, Germany, Italy and Spain. Meanwhile, another study showed the importance of nitrogen dioxide along with population density in determining the coronavirus pandemic rate in England. In this follow-up study, Aerosol Optical Depth (AOD) was introduced in conjunction with other variables like nitrogen dioxide and population density for further analysis in fifty-four administrative regions of Germany, Italy and Spain. The AOD values were extracted from the Moderate Resolution Imaging Spectroradiometer (MODIS) onboard the Terra and Aqua satellites while the nitrogen dioxide data were extracted from TROPOMI (TROPOspheric Monitoring Instrument) sensor onboard the Sentinel-5 Precursor satellite. Regression models, as well as multiple statistical tests were used to evaluate the predictive skill and significance of each variable to the fatality rate. The study was conducted for two periods: (1) pre-exposure period (Dec 1, 2019-Feb 29, 2020); (2) complete exposure period (Dec 1, 2019-Jul 1, 2020). Some of the results pointed towards AOD potentially being a factor in estimating the coronavirus fatality rate. The models performed better using the data collected during the complete exposure period, which showed higher AOD values contributed to an increased significance of AOD in the models. Meanwhile, some uncertainties of the analytical results could be attributed to data quality and the absence of other important factors that determine the coronavirus fatality rate.


A glycoprotein multimer from Bacillus thuringiensis sporangia: dissociation into subunits and sugar composition.
Two glycoproteins (205 and 72 kDa) were found in Bacillus thuringiensis sporangia. They were predominantly localized in the exosporium and/or the spore coat, although a small proportion was also found in membranes. A method for the dissociation of hydrophobic aggregates that resist the usual conditions of SDS-PAGE is described. Using this method we established that the 205 kDa glycoprotein is a multimer of the 72 kDa one. Deglycosylation of the 205 kDa and 72 kDa glycoproteins with trifluoromethanesulfonic acid yielded a 54 kDa polypeptide in both cases. At least three species of oligosaccharides were O-glycosidically linked to serines of the 54 kDa polypeptide chain. One of the oligosaccharides had N-acetylgalactosamine at the reducing end, rhamnose and a component not yet identified.


The road to zero preventable birth injuries.
The Seton Family of Hospitals' experience in developing and implementing transformational practices in labor and delivery (L&D) units aimed at reducing the rate of birth trauma at our facilities was previously reported. Seton began its individual perinatal safety effort in earnest in October 2003. The endeavor brought together the four hospitals that offer obstetrical services, resulting in the establishment of an interdisciplinary team. The team meets monthly to develop and monitor best practices that are then shared, executed, and validated by each respective site's perinatal councils. A 36% reduction in the use of vacuum and forceps (from a frequency of 7.4% to 4.7%) was previously reported; the current rate (fiscal year [FY] 2007-FY 2008 year-to-date; July 1, 2006-December 31, 2007) was 4.1%. The incidence of associated birth trauma decreased to 0% for the last 15 months (ending December 31, 2007). During the first three project years (FY 2004-FY 2006), the average length of stay for infants admitted to the neonatal intensive care unit for birth injury declined by 80% (as compared with the previous three years), from 15.8 to 3.1 days. The perinatal safety team developed processes that have resulted in large and sustained reductions in the rate of serious birth trauma at all obstetric facilities.


Complementary ADF-STEM: a Flexible Approach to Quantitative 4D-STEM.
Scanning transmission electron microscopy (STEM) has a broad range of applications in materials characterization, including real-space imaging, spectroscopy, and diffraction, at length scales from the micron to sub-Ångström. The recent development and adoption of high-speed, direct electron STEM detectors has enabled diffraction patterns to be collected at each probe position, generating four-dimensional STEM (4D-STEM) datasets and opening new imaging modalities. However, the limited pixel numbers in these detectors enforce a tradeoff between angular resolution and maximum collection angle. In this paper, we describe a straightforward method for quantifying 4D-STEM data by utilizing the full flux of the electron beam, including electrons scattered beyond the limits of the detector. This enables significantly increased experimental flexibility, including the synthesis of quantitative, high-contrast complementary annular dark field (cADF) STEM images from low-angle diffraction patterns whilst maintaining high angular resolution; as well as the optimization of electron dose and the more effective use of low dynamic range detectors.


Bioaccumulation kinetics of the organochlorine pesticide mirex in amphipods.
The survival of Hyallela azteca was found to be reduced relative to that of Crangonyx pseudogracilis during exposure to mirex in water for a 13-day period. This phenomenon appeared to be correlated to greater bioaccumulation of mirex by H. azteca than by C. pseudogracilis. H. azteca was found to exhibit two-compartment kinetics, while C. pseudogracilis was described by single-compartment kinetics. The bioaccumulation differences between the two species were the result of both uptake and clearance differences: uptake into a "fast" compartment of H. azteca, greater uptake into the slow compartment of H. azteca relative to the single compartment of C. pseudogracilis, and greater clearance from the single C. pseudogracilis compartment relative to the slow compartment of H. azteca.


Effect of aspirin and dipyridamole treatment on prostacyclin production by human veins.
Patients admitted for surgical removal of varicose veins were treated in a blinded manner for 48 hours prior to surgery with either placebo, low-dose aspirin (25 mg twice daily), dipyridamole (150 mg twice daily) or both. Segments of vein excised at surgery were incubated with or without sodium arachidonate and subsequent prostacyclin (PGI2) production was measured without knowledge of treatment given. During the first 5 minute period of incubation in the presence of arachidonate, veins from dipyridamole-treated patients demonstrated increased (by 75%) arachidonate-stimulated PGI2 production compared to placebo-treated patients. By contrast, PGI2 production was reduced by 64% by aspirin treatment and 67% by aspirin plus dipyridamole compared to placebo-treated patients (p = less than 0.05). In unstimulated vein segments incubated in the absence of arachidonate, spontaneous PGI2 production during the first 5 minute incubation period was increased 32% following dipyridamole treatment but was unchanged following aspirin treatment. By contrast, unstimulated (spontaneous) PGI2 production in patients treated with aspirin plus dipyridamole was reduced by 57% (p = less than 0.05), compared to both placebo- and aspirin-treated patients, and by 71% (p = less than 0.05) compared to dipyridamole-treated patients. With repeated change of incubation medium, the ability of vein walls to produce PGI2 declined. This exhaustion was not prevented by drug treatment. However, drug effects between patient treatment groups were consistent over successive incubation periods. These results suggest that certain therapeutic benefits that might be achieved by enhancement of PGI2 production from vascular endothelium following dipyridamole treatment may be reduced by simultaneous aspirin treatment.(ABSTRACT TRUNCATED AT 250 WORDS)


Colostomies--indications and contraindications: Lahey Clinic experience, 1963--1974.
Over a 12-year period, 276 temporary colostomies were performed on 271 patients. During this interval 118 colostomies were closed. The morbidity rate of colostomy construction was 21 per cent, and for closure it was 49.1 per cent. No definite factor could be indentified as contributing to this high rate of complications for colostomy construction. With respect to colostomy closure, predisposing factors that seemed to increase morbidity were shorter interval between creation and closure of the stoma and resection of colostomy (as opposed to closure without resection). Intra-abdominal drains were associated with a prohibitively high rate of wound infection, although subcutaneous drainage was not successful in reducing the incidence of infection significantly.


In DA Club of Reinforcement: Glutamate, It's Your Birthday.
Dopamine release guides reward encoding, but the contribution of glutamate remains unclear. In this issue of Neuron, Zell et al. leverage the genetic ablation of dopamine synthesis from midbrain VGluT2 neurons to assess how glutamate shapes positive reinforcement.


Endothelial function progressively deteriorates during normal pregnancy.
To elucidate changes in endothelial function throughout the gestational period in normal pregnancy and its relationship with plasma soluble fms-like tyrosine kinase-1 (sFlt-1) levels. Endothelial function was evaluated by reactive hyperemia index (RHI) using Endo-PAT2000 and plasma sFlt-1 levels were measured simultaneously by ELISA. RHI gradually deteriorated with increasing gestational age. Plasma sFlt-1 levels exhibited a gradual increase at late pregnancy and were inversely correlated with RHI. Maternal endothelial function gradually deteriorates with increasing gestational age and there is an inverse correlation between endothelial function and plasma sFlt-1 levels in normal pregnancy.


Somatization: a psychiatrist's perspective.
Psychiatrists are referred only a small and unrepresentative proportion of patients who somatize. To a large extent this is due to the fact that patients consider themselves physically ill and regard psychiatric referral as inappropriate. Although somatization has several social advantages it obscures underlying psychopathology and leads to unnecessary physical investigations with a risk of iatrogenic disease. It is important to establish a psychiatric diagnosis as early as possible so that treatment can be started before a chronic pattern of illness behaviour is established.


New VAPB deletion variant and exclusion of VAPB mutations in familial ALS.
Amyotrophic lateral sclerosis (ALS) is a progressive, neurodegenerative disorder involving upper and lower motor neurons. The vesicle-associated membrane protein B (VAPB) gene has been genetically linked to ALS in several large Brazilian families in which the disorder is caused by a proline to serine mutation at codon 56 (P56S). No additional mutations have been identified. To establish the prevalence of VAPB mutations, we screened 80 familial ALS samples by DNA sequencing. Our study failed to identify any novel VAPB gene mutations but identified a single Brazilian family harboring the P56S mutation. In a second familial ALS case, we identified a three-base pair deletion within exon 5 of the VAPB gene that deleted the serine residue at position 160 (Delta S160). This variant is detected in a normal population at low frequency (0.45%). Analyses of homology alignment and secondary structure predict that this deletion significantly alters the structure of VAPB, although a GFP-Delta S160 VAPB fusion protein demonstrates a wild-type subcellular localization. This contrasts the aberrant localization observed in a GFP-P56S VAPB fusion protein. The allele frequency of Delta S160 in patients with sporadic ALS does not differ significantly from that in the normal population. Mutations in the VAPB gene are rare and the Delta S160 variant does not contribute to the development of amyotrophic lateral sclerosis.


Influence of different types of pulp treatment during isolation in the obtention of human dental pulp stem cells.
Different methods have been used in order to isolate dental pulp stem cells. The aim of this study was to study the effect of different types of pulp treatment during isolation, under 3% O2 conditions, in the time needed and the efficacy for obtaining dental pulp stem cells. One hundred and twenty dental pulps were used to isolate dental pulp stem cells treating the pulp tissue during isolation using 9 different methods, using digestive, disgregation, or mechanical agents, or combining them. The cells were positive for CD133, Oct4, Nestin, Stro-1, CD34 markers, and negative for the hematopoietic cell marker CD-45, thus confirming the presence of mesenchymal stem cells. The efficacy of dental pulp stem cells obtention and the minimum time needed to obtain such cells comparing the 9 different methods was analyzed. Dental pulp stem cells were obtained from 97 of the 120 pulps used in the study, i.e. 80.8% of the cases. They were obtained with all the methods used except with mechanical fragmentation of the pulp, where no enzymatic digestion was performed. The minimum time needed to isolate dental pulp stem cells was 8 hours, digesting with 2mg/ml EDTA for 10 minutes, 4mg/ml of type I collagenase, 4mg/ml of type II dispase for 40 minutes, 13ng/ml of thermolysine for 40 minutes and sonicating the culture for one minute. Dental pulp stem cells were obtained in 97 cases from a series of 120 pulps. The time for obtaining dental pulp stem cells was reduced maximally, without compromising the obtention of the cells, by combining digestive, disgregation, and mechanical agents.


Midfacial protraction with skeletally anchored face mask therapy: a novel approach and preliminary results.
The purpose of this pilot study was to investigate the effects of facemask treatment with a rigid skeletal anchor. Miniplates were placed on the lateral nasal wall of the maxilla in 6 subjects (mean age, 11.8 +/- 1.1 years). No dental tissue was incorporated into the anchorage system. Lateral cephalograms of each patient were evaluated at the beginning of treatment (T1), at the end of active treatment (T2), and at the end of follow-up (T3). In the horizontal plane, mean A-point movement was 4.8 +/- 2.0 mm in a mean of 10.8 +/- 2.4 months. The infraorbital region showed significant anterior movement (mean, 3.3 +/- 1.1 mm) at T2. At T3, advancement of the maxilla and the infraorbital region was well maintained (mean follow-up time, 15.2 +/- 0.9 months). Miniplate stability was excellent in all patients throughout treatment. Remarkable advancement in the middle face and consequent fullness in the soft-tissue profile can be achieved by using skeletal anchorage in conjunction with facemask therapy in the late mixed-dentition period.


Evaluation of MR in spinal dysraphism with lipoma: comparison with metrizamide computed tomography.
Magnetic resonance (MR) imaging and metrizamide computed tomography (CT) were performed on three patients with spinal dysraphism with lipoma. Sagittal MR images were superior to metrizamide CT reformatted sagittal images in demonstrating anatomic relationships of the spinal cord, filum terminale and lipoma. In the axial plane, CT proved superior to MR. In all cases, the T1 weighted images with exceptional spatial resolution and excellent fat discrimination were more helpful than T2 weighted images. Although both MR and CT are useful in the initial evaluation of these disorders, subsequent evaluation employing only MR is probably adequate. Promising developments in surface coil imaging may eventually replace CT.


Nodular lymphoid hyperplasia of terminal ileum: how to avoid overdiagnosis of Crohn's terminal ileitis in MR enterography?
To investigate the magnetic resonance enterography (MRE) characteristics of nodular lymphoid hyperplasia (NLH) and Crohn's terminal ileitis (CTI). Of 1552 MREs from November 2011 to July 2018, 61 individuals with biopsy-proven NLH (n = 24) and CTI (n = 37, 27 with active CTI) were selected based on the inclusion criteria. NLH cases were also followed up for median (range) of 40 (21-61) months. Two board-certified radiologists, blind to clinical data and diagnosis, reviewed MRE in consensus. Conventional, morphological, enhancement, and diffusion parameters were assessed. Mann-Whitney, χ2, and logistic regression analyses were conducted. No NLH patient developed inflammatory bowel disease or lymphoproliferative disorders during the follow-up. Serosal surface irregularity (65% vs. 8%), pseudo-diverticula (27% vs. 0), and mesenteric fat involvement (38% vs. 4%) were more frequent in CTI than NLH (p < 0.01), while mucosal nodularity was more prevalent in NLH (71%) than CTI (19%) (p < 0.001). The upstream luminal diameter (15.0 vs. 12.5 mm, p = 0.015) and mural thickness (6.0 vs. 4.0 mm, p < 0.001) of the terminal ileum showed higher values in CTI than NLH. Unlike enhancement and diffusion parameters, morphological features (mucosal nodularity, serosal surface irregularity, and mesenteric fat involvement) could distinguish NLH from CTI regardless of CTI activity.


Antibodies against synthetic peptides as a tool for functional analysis of the transforming protein pp60src.
To study the function of pp60src, the transforming protein encoded for by Rous sarcoma virus, we have raised antibodies against synthetic oligopeptides corresponding to the primary structure of pp60src. All eight investigated peptides were immunogenic in rabbits, and four induced pp60src-specific antibodies. We screened tumor-bearing rabbit (TBR) sera for antibodies against the peptides; this revealed that five out of six of the peptides, chosen according to a high hydrophilicity plot, were related to epitopes of native pp60src, in contrast to two peptides of low hydrophilicity, which contained a cleavage site for protease. Antibodies against three of the peptides appeared to react with the kinase-active site of pp60src, as these antibodies were phosphorylated in their heavy chain upon immune precipitation. Antibodies against two of the peptides, in contrast to the others, did not precipitate pp60src when this molecule was complexed with two cellular proteins, pp50 and pp90. This observation allows speculation about the location of the pp60src site involved in the formation of this complex.


The effect of optical degradation on the contrast sensitivity function measured at the fovea and in the periphery.
Contrast sensitivity functions (CSF's) were measured for a zone of 0.5 degree angular radius at the fovea and for annular zones extending from 0.5 degrees eccentricity to 1 degree, from 1 to 2 degrees and from 2 to 4 degrees. The effect of optical degradation was taken into account by the application of a correction factor determined from the Modulation Transfer Function (MTF). Using the reciprocal of the peak of the CSF [1/SF (max)] as a parameter, the effect of applying the MTF correction on the CSF with eccentricity was determined. The effect of applying an MTF correction is significantly to increase the rate of change of 1/SF (max) with eccentricity. Since it has been reported that hyperacuity is relatively unaffected by optical degradation, it is proposed that the differing effects of optical degradation between grating acuity and hyperacuity may account, at least partially, for the different rates of decline of these thresholds with eccentricity. The implications of this proposal for M-scaling theories and strategies are considered.


Lexical decision tasks in depressive patients: semantic priming before and after clinical improvement.
This study was designed to evaluate the effect of semantic priming with a lexical decision task in 22 depressed patients (DSM-III-R, 1987) and 30 control subjects. These patients were evaluated twice: first when they arrived at the hospital, and secondly, after clinical improvement. Clinical improvement was evaluated using standard depression rating scales. A lexical decision task involving semantic relations (related vs. unrelated, e.g., apple-pear) was used to evaluate the processing of semantic information. The results showed that, for the first evaluation, the depressives presented similar semantic priming to control subjects. When we compared semantic priming in the first and the second passes, we observed that its amplitude was identical. The sole difference between the two passes concerns the global reaction time in the depressive group. This last result suggested that, with clinical improvement, the characteristic psychomotor retardation declines. One of the major results concerns the fact that severe depressive patients (first pass) exhibit normal semantic priming in a lexical decision task. These results indicate, in this clinical population, the preservation of controlled processes implicated in this lexical decision task.


The denaturation of DNA.
Nucleic acids display a very characteristic ultraviolet absorption spectrum with a maximum near 260 nm and a minimum near 230 nm. It was discovered in 1950 that the spectrum of native DNA is much lower than the spectrum calculated from its constituent nucleotides (nt). The observed absorbance strongly increases (hyperchromic effect), not only following hydrolysis, but also following mild treatments, known to respect the phosphoester bonds between nt (and to have little effect on the molecular weight). These results show that DNA has a secondary structure made of secondary valence interactions between the nitrogen bases, and that this secondary structure collapses (DNA denaturation) following mild treatments (low pH, high temperature or, even simply, low salt concentration).


Enzyme-accelerated and structure-guided crystallization of calcium carbonate: role of the carbonic anhydrase in the homologous system.
The calcareous spicules from sponges, e.g. from Sycon raphanus, are composed of almost pure calcium carbonate. In order to elucidate the formation of those structural skeletal elements, the function of the enzyme carbonic anhydrase (CA), isolated from this species, during the in vitro calcium carbonate-based spicule formation, was investigated. It is shown that the recombinant sponge CA substantially accelerates calcium carbonate formation in the in vitro diffusion assay. A stoichiometric calculation revealed that the turnover rate of the sponge CA during the calcification process amounts to 25 CO2s(-1) × molecule CA(-1). During this enzymatically driven process, initially pat-like particles are formed that are subsequently transformed to rhomboid/rhombohedroid crystals with a dimension of ~50 μm. The CA-catalyzed particles are smaller than those which are formed in the absence of the enzyme. The Martens hardness of the particles formed is ~4 GPa, a value which had been determined for other biogenic calcites. This conclusion is corroborated by energy-dispersive X-ray spectroscopy, which revealed that the particles synthesized are composed predominantly of the elements calcium, oxygen and carbon. Surprising was the finding, obtained by light and scanning electron microscopy, that the newly formed calcitic crystals associate with the calcareous spicules from S. raphanus in a highly ordered manner; the calcitic crystals almost perfectly arrange in an array orientation along the two opposing planes of the spicules, leaving the other two plane arrays uncovered. It is concluded that the CA is a key enzyme controlling the calcium carbonate biomineralization process, which directs the newly formed particles to existing calcareous spicular structures. It is expected that with the given tools new bioinspired materials can be fabricated.


The development and validation of a psychological contract of safety scale.
This paper builds on previous research by the author and describes the development and validation of a new measure of the psychological contract of safety. The psychological contract of safety is defined as the beliefs of individuals about reciprocal safety obligations inferred from implicit and explicit promises. A psychological contract is established when an individual believes that perceived employer and employee safety obligations are contingent on each other. A pilot test of the measure is first undertaken with participants from three different occupations: nurses, construction workers, and meat processing workers (N=99). Item analysis is used to refine the measure and provide initial validation of the scale. A larger validation study is then conducted with a participant sample of health care workers (N=424) to further refine the measure and to determine the psychometric properties of the scale. Item and correlational analyses produced the final employer and employee obligations scales, consisting of 21 and 17 items, respectively. Factor analyses identified two underlying dimensions in each scale comparable to that previously established in the organizational literature. These transactional and relational-type obligations provided construct validity of the scale. Internal consistency ratings using Cronbach's alpha found the components of the psychological contract of safety measure to be reliable. The refined and validated psychological contract of safety measure will allow investigation of the positive and negative outcomes associated with fulfilment and breach of the psychological contract of safety in future research.


Signature-tagged and directed mutagenesis identify PABA synthetase as essential for Aspergillus fumigatus pathogenicity.
Signature-tagged mutagenesis (STM) is a method that has been used to screen for genes required for in vivo survival of pathogenic bacteria, but has not been used to investigate a eukaryotic pathogen in an animal model of disease. We have adapted STM to identify genes required for in vivo growth of the opportunistic fungal pathogen Aspergillus fumigatus. Using a mouse model of invasive pulmonary aspergillosis, we have isolated several mutant strains with defects in their ability to replicate in vivo. One strain unable to cause lethal infection was further characterized and found to have an insertion into the promoter of a gene (pabaA) encoding para-aminobenzoic acid synthetase, an enzyme catalyzing a late step in the biosynthesis of folate. The complete inability of this strain, and other pabaA- strains constructed in this study by targeted gene deletion, to cause lethal infection in mice confirms the importance of the folate synthesis pathway for in vivo survival of this pathogen. The successful application of STM to A. fumigatus demonstrates that in vivo genetic analysis of eukaryotic pathogens is feasible and could result in the identification of potential targets, such as para-aminobenzoic acid synthetase, for novel antifungal therapies.


Cerebral glucose utilization: local changes during and after recovery from spreading cortical depression.
Cerebral glucose utilization is markedly increased in most areas of the cerebral cortex and reduced in many subcortical structures during spreading cortical depression. During recovery, cortical glucose utilization is still elevated, but the increased metabolic activity is distributed in columns running perpendicularly through the cortex.


"Skull punch" for craniotomy in neonates and infants.
We have designed a new instrument that we have called a "skull punch" which is useful and safe for making a burr hole for a purely osteoplastic craniotomy in neonates and infants. The instrument is described in detail.


Family structure and mental health: the mediating effects of socioeconomic status, family process, and social stress.
Although numerous studies reveal differences in mental health by the structure of one's family of origin, there remains debate regarding the processes generating these patterns. Using a sample of young adults (19-21 years) in Miami-Dade County in Florida, this study examines the explanatory significance of three presumed correlates of family type: socioeconomic status, family processes, and level of social stress. Consistent with prior research, our results reveal higher levels of depressive symptoms among those from stepfamilies, single parent families, and single parent families with other relatives present, compared with mother-father families. All three presumed correlates make significant independent contributions to the prediction of depressive symptomatology. Substantial mediating effects also are observed for all three explanatory dimensions. Collectively, they completely or largely explain observed family type variations in mental health risk.


A tumor suppressor function of Smurf2 associated with controlling chromatin landscape and genome stability through RNF20.
In addition to allelic mutations, cancers are known to harbor alterations in their chromatin landscape. Here we show that genomic ablation of Smad ubiquitin regulatory factor 2 (Smurf2), a HECT-domain E3 ubiquitin ligase, results in dysregulation of both the DNA damage response and genomic stability, culminating in increased susceptibility to various types of cancers in aged mice. We show that Smurf2 regulates the monoubiquitination of histone H2B as well as the trimethylation of histone H3 at Lys4 and Lys79 by targeting ring finger protein 20 (RNF20) for proteasomal degradation in both mouse and human cells. We also show that Smurf2 and RNF20 are colocalized at the γ-H2AX foci of double-stranded DNA breaks in the nucleus. Thus, Smurf2 has a tumor suppression function that normally maintains genomic stability by controlling the epigenetic landscape of histone modifications through RNF20.


Cerebellar glioblastoma: a clinical series with contemporary molecular analysis.
Glioblastomas (GBM) are localized in only less than 1% of patients in the cerebellum. Therefore, tumor characteristics, survival, and the efficacy of therapies are not yet well defined. The present study aims to characterize the molecular features of cerebellar GBM (GBMc) in 8 patients treated with contemporary modality in our institution. Patients' treatment history, progression-free survival (PFS), and overall survival (OS) were analyzed. All histopathological specimens were re-investigated. EGFR amplification was determined by FISH, H3F3A, and HIST1H3B mutation status and MGMT promoter methylation after bisulfite treatment by pyrosequencing and BRAF V600E by pyrosequencing and immunohistochemistry. TERT promoter mutations were analyzed by Sanger sequencing, CDKN2A/B deletions by digital PCR. The expression of IDH1 R132H, ATRX, and p53 was determined through immunohistochemistry. Six adults and two children (mean age 36 years) underwent tumor resection via medial or lateral suboccipital craniotomy. The median overall survival (mOS) of the adult patients was 7 months. GBMc from two children demonstrated a H3F3A K27M mutation. One of these also harbored a BRAF V600E mutation and has already had a PFS of 74 months. Mutated IDH1 R132H protein was expressed in 2 GBM from adult patients with previous supratentorial anaplastic astrocytoma. One patient carried a TERT promoter mutation. Another patient initially presented with a thalamic pilocytic astrocytoma. The cerebellar tumor revealed neither a BRAF V600E nor a H3F3A mutation but a homozygous CDKN2A/B deletion. GBM located in the cerebellum can be found in all age groups. We provide novel molecular genetic data on these rare tumors. Mutated IDH1 R132H protein and H3F3A K27M mutations indicate that a substantial number of GBMc are "metastatic" or "diaschismatic" lesions. Mutation of BRAF V600E may have a stronger biological significance than H3F3A K27M alterations. In a subset of patients, GBM may arise primarily as a distinct entity in the cerebellum.


Antitumor effect and safety profile of systemically delivered oncolytic adenovirus complexed with EGFR-targeted PAMAM-based dendrimer in orthotopic lung tumor model.
Adenovirus (Ad)-mediated cancer gene therapy has been proposed as a promising alternative to conventional therapy for cancer. However, success of systemically administered naked Ad has been limited due to the immunogenicity of Ad and the induction of hepatotoxicity caused by Ad's native tropism. In this study, we synthesized an epidermal growth factor receptor (EGFR)-specific therapeutic antibody (ErbB)-conjugated and PEGylated poly(amidoamine) (PAMAM) dendrimer (PPE) for complexation with Ad. Transduction of Ad was inhibited by complexation with PEGylated PAMAM (PP) dendrimer due to steric hindrance. However, PPE-complexed Ad selectively internalized into EGFR-positive cells with greater efficacy than either naked Ad or Ad complexed with PP. Systemically administered PPE-complexed oncolytic Ad elicited significantly reduced immunogenicity, nonspecific liver sequestration, and hepatotoxicity than naked Ad. Furthermore, PPE-complexed oncolytic Ad demonstrated prolonged blood retention time, enhanced intratumoral accumulation of Ad, and potent therapeutic efficacy in EGFR-positive orthotopic lung tumors in comparison with naked Ad. We conclude that ErbB-conjugated and PEGylated PAMAM dendrimer can efficiently mask Ad's capsid and retarget oncolytic Ad to be efficiently internalized into EGFR-positive tumor while attenuating toxicity induced by systemic administration of naked oncolytic Ad.


Correlation of Structure and Dynamics Behavior in Polyzwitterions: From Concentrated Solution to Gel-Like State.
The dynamic behaviors of polyzwitterions, poly(4-((3-methacrylamidopropyl) dimethylammonio) butane-1-sulfonate) (PSBP), are investigated using dynamic light scattering, small angle X-ray scattering, and rheology. The findings reveal two relaxation modes, including a fast and a slow mode, which are observed in both solution state and gel-like state, with varying polyzwitterion concentration (CP ) and NaCl concentration (CNaCl ). As CP and CNaCl increasing, a slower slow mode and a faster fast mode are observed. The fast mode corresponds to the diffusion of chains, while the slow mode arises from chain aggregations. In solutions, the slow mode is dominated by the diffusion of chain aggregations. However, in the gel-like state, the "cage network" traps aggregations more densely, leading to their dynamic behavior being dominated by enhanced topological entanglements and ionic interactions. This difference highlights the unique nature of the slow relaxation mode between concentrated solution and gel-like state, arising from changes in the average distance between chain aggregations resulting from increased CP and CNaCl concentrations.


Galanin differentially regulates acetylcholine release in ventral and dorsal hippocampus: a microdialysis study in awake rat.
The purpose of the present study was to investigate, by use of in vivo microdialysis technique, the regulatory role of galanin on acetylcholine (ACh) release in the CA1, CA3, and dentate gyrus (DG) subregions of rat dorsal and ventral hippocampus. In the ventral hippocampus, local infusions of galanin (1.5 nmol) into CA1, and CA3, but not DG (3 nmol), decreased basal ACh release to 58.6% and 68.4%, respectively. In addition, local infusion of galanin (1.5 nmol) into the ventral DG, and CA3 areas decreased basal ACh levels in the CA1 to 51.2% and 84%, respectively. This observation implies that the effects of galanin are unlikely to be mediated via galanin autoreceptors on the cholinergic terminals, but rather via mechanisms involving galanin internalization and modulation of hippocampo-septo-hippocampal loops, attenuation of the excitability of the principal cells, or indirect modulation by galanin-containing vasopressin terminals to the ventral and/or dorsal hippocampus. In the dorsal hippocampus, galanin infusion (1.5 nmol) into the CA1 region increased ACh release to 128.2% of the control levels, but infusions of galanin had no effects in the CA3 and DG. In all cases, the ACh levels returned to basal values within 100 min after the galanin infusion. It is concluded that the attenuating effects of galanin on ACh release in the ventral hippocampus and increase in ACh release in the dorsal hippocampus are in line and support the current view on molecular and functional distinction between the ventral hippocampus being involved preferentially in motivational and emotional behavior, whereas the dorsal hippocampus is primarily implicated in cognitive processes of learning and memory.


Edge-modification of laponite with dimethyl-octylmethoxysilane.
This study examines the edge modification of laponite, with a monoalkoxy silane, dimethyl-octylmethoxysilane. The influence of ultrasonics, aging time and silane concentration on the resultant materials is examined. The silylated clays are characterized by XRD, IES, TGA, and Si NMR. The amount of grafted silane was increased by increasing the ratio of silane to clay, sonication of the reaction mixture and ageing the reaction mixture for no more than 24 h to avoid removal of grafted silane due to equilibrium effects.


Calmodulin-dependent protein kinase and associated substrates in Torpedo electric organ.
Calcium plus calmodulin (Ca2+/CaM)-dependent protein kinase activity was demonstrated in subcellular fractions from Torpedo californica electric organ. A protein kinase activity dependent on Ca2+/CaM was purified about 200-fold from electric organ cytosol using DEAE-cellulose and CaM-affinity chromatography. The most effective exogenous substrates for this enzyme were the synapse-specific protein Synapsin I (Protein I) and histone f3. Phosphorylase b, skeletal muscle myosin light chains, casein, phosvitin, histone f2b, and G-substrate were relatively poorly phosphorylated by Torpedo CaM-protein kinase. Thus, the enzyme differs in its substrate specificity from known cyclic nucleotide-dependent protein kinases, myosin light chain kinase and phosphorylase kinase. The Km for ATP was 15-20 microM; for Synapsin I, 0.8 microM; and for CaM, 85 nM. Two major endogenous substrates (Mr = 62,000 and 54,000) for CaM-protein kinase co-purified with the enzyme through the CaM-affinity column step. These two substrates, as well as the enzyme, were present in other subcellular fractions in addition to the cytosol, including crude membranes and purified synaptic vesicles. A third major substrate (Mr = 39,000) could be separated from the enzyme during purification and appeared to be localized primarily in the cytosol. CaM-protein kinase increased the phosphorylation of both serine and threonine residues in endogenous substrates. In contrast to previous reports, no evidence for Ca2+/CaM-dependent phosphorylation of any subunit of the acetylcholine receptor was obtained.


A kinetics study on intercalation pseudocapacitance of layered TiS2 in K-ion batteries.
The boundary between a capacitor and a battery is vague for layered transition metal sulfides, especially when it comes to TiS2 with a multi-phase transition process. In this work we analyzed both the quasi-static process and dynamic process of K+ storage in TiS2 with carbonate based electrolytes, which proved to be an intercalation pseudocapacitive behavior.


Traumatic amputation of the left lower renal pole in children.
Four children between 5 and 10 years old suffered traumatic amputation of the left lower renal pole following flank trauma. All patients were evaluated with excretory urography and isotope renography. The renal scan clearly demonstrated failure of perfusion of the lower renal pole and urinary extravasation, and was believed to be more valuable than the standard excretory urogram as a diagnostic tool. All children were managed similarly: delayed (72 to 96 hours) exploration, simple removal of the amputated segment and insertion of a Penrose drain. They all have done well. The patients were normotensive at followup and had excellent function of the remaining portion of the kidney.


Intracellular reactive oxygen species trigger mitochondrial dysfunction and apoptosis in cadmium telluride quantum dots-induced liver damage.
Quantum dots (QDs), also known as semiconductor QDs, have specific photoelectricproperties which find application in bioimaging, solar cells, and light-emitting diodes (LEDs). However, the application of QDs is often limited by issues related to health risks and potential toxicity. The purpose of this study was to provide evidence regarding the safety of cadmium telluride (CdTe) QDs by exploring the detailed mechanisms involved in its hepatotoxicity. This study showed that CdTe QDs can increase reactive oxygen species (ROS) in hepatocytes after being taken up by hepatocytes, which triggers a significant mitochondrial-dependent apoptotic pathway, leading to hepatocyte apoptosis. CdTe QDs-induce mitochondrial cristae abnormality, adenosine triphosphate (ATP) depletion, and mitochondrial membrane potential (MMP) depolarization. Meanwhile, CdTe QDs can change the morphology, function, and quantity of mitochondria by reducing fission and intimal fusion. Importantly, inhibition of ROS not only protects hepatocyte viability but can also interfere with apoptosis and activation of mitochondrial dysfunction. Similarly, the exposure of CdTe QDs in Institute of Cancer Research (ICR) mice showed that CdTe QDs caused oxidative damage and apoptosis in liver tissue. NAC could effectively remove excess ROS could reduce the level of oxidative stress and significantly alleviate CdTe QDs-induced hepatotoxicity in vivo. CdTe QDs-induced hepatotoxicity may originate from the generation of intracellular ROS, leading to mitochondrial dysfunction and apoptosis, which was potentially regulated by mitochondrial dynamics. This study revealed the nanobiological effects of CdTe QDs and the intricate mechanisms involved in its toxicity at the tissue, cell, and subcellular levels and provides information for narrowing the gap between in vitro and in vivo animal studies and a safety assessment of QDs.


Inappropriate Prescribing of Antibiotics to Pediatric Patients Receiving Medicaid: Comparison of High-Volume and Non-High-Volume Antibiotic Prescribers-Kentucky, 2019.
Inappropriate antibiotic prescribing to pediatric Medicaid patients was compared among high-volume and non-high-volume prescribers. High-volume prescribers had a higher percentage of inappropriate prescriptions than non-high-volume prescribers (17.2% versus 15.8%, p = 0.005). Targeting high-volume prescribers for stewardship efforts is a practical approach to reducing outpatient antibiotic prescribing that also captures inappropriate use.


Negishi cross-coupling enabled synthesis of novel NAD(+)-dependent DNA ligase inhibitors and SAR development.
Two novel compounds, pyridopyrimidines (1) and naphthyridines (2) were identified as potent inhibitors of bacterial NAD(+)-dependent DNA ligase (Lig) A in a fragment screening. SAR was guided by molecular modeling and X-ray crystallography. It was observed that the diaminonitrile pharmacophore made a key interaction with the ligase enzyme, specifically residues Glu114, Lys291, and Leu117. Synthetic challenges limited opportunities for diversification of the naphthyridine core, therefore most of the SAR was focused on a pyridopyrimidine scaffold. The initial diversification at R(1) improved both enzyme and cell potency. Further SAR developed at the R(2) position using the Negishi cross-coupling reaction provided several compounds, among these compounds 22g showed good enzyme potency and cellular potency.


Does Mode of Survey Administration Matter? Using Measurement Invariance to Validate the Mail and Telephone Versions of the Bereaved Family Survey.
The Veterans Health Administration evaluates outcomes of end-of-life (EOL) care using the Bereaved Family Survey (BFS). Originally, the BFS was administered as a telephone survey but was transitioned to a mail survey beginning October 2012. The transition necessitated an evaluation of the tool's validity using this new mode of administration. The objective of this study was to validate the mail version and to test for measurement invariance (MI) across the two administration modes. Telephone and mail versions of the BFS were validated separately between October 2009 and September 2013. MI was evaluated using a series of confirmatory factor analyses (CFAs). Construct validity was evaluated by calculating Cronbach alpha coefficients and examining differences between BFS factor scores for groups with and without quality care indicators (e.g., receipt of a palliative care consult). Our sample consisted of 35,682 decedent BFS scores (27,109 telephone surveys; 8573 mail surveys). BFS item scores were slightly skewed, with a predominance of higher scores for both the telephone and mail version. The average missing rate for each BFS item was minimal, just 2% for each version. The CFA models demonstrated dimensional, configural, metric, and factor mean invariance across administration modes. BFS factor scores were consistently higher when a patient received EOL quality care indicators regardless of mode of administration. These findings demonstrate the MI and robust psychometric properties for the BFS across administration modes.


Cooled-tip ablation results in increased radiofrequency power delivery and lesion size in the canine heart: importance of catheter-tip temperature monitoring for prevention of popping and impedance rise.
Since myocardial lesion size during radio-frequency (RF) ablation is limited at high power by impedance rise when electrode tip temperature exceed 100 degrees C, controlling tip temperature by continuous intraelectrode saline infusion could permit generation of larger lesion. (1) Two dogs randomly received either standard or cooled tip RF ablation at 4 to 6 separate LV sites. Power output of 30 W was delivered via modified 7 Fr deflectable catheter with 4 mm tip for up to 120 sec or until impedance rise occurred. (2) Six dogs randomly received cooled tip RF ablation at power output of 20, 30, 40 W for 120 sec. (3) Three dogs randomly received cooled tip RF ablation using room temperature saline (21-25 degrees C) or chilled saline (1-4 degrees C) infusion. Overall, peak tip temperature was lower for cooled vs standard RF deliveries (97 +/- 17 degrees C vs. 42 +/- 8 degrees C). Lesion depth and volume were significantly larger for cooled burns. Lesion depth and volume and the incidence of abrupt impedance rise/popping did not differ between room temperature saline and chilled saline infusion. The catheter-tip temperature at the onset of popping and abrupt impedance rise was 54 +/- 5 degrees C (48-60 degrees C) and 59 +/- 10 degrees C (50-75 degrees C). Cooled tip RF current delivery at high power is associated with increased myocardial lesion size which may facilitate successful ablation of ventricular tachycardia associated with acquired structural heart disease. Catheter-tip temperature should be maintained below 45 degrees C to prevent popping and abrupt impedance rise during RF energy delivery.


Covering bariatric surgery has minimal effect on insurance premium costs within the Affordable Care Act.
Currently, of the 51 state health exchanges operating under the Affordable Care Act, only 23 include benchmark plans that cover bariatric surgery coverage. Bariatric surgery coverage is not considered an essential health benefit in 28 state exchanges, and this lack of coverage has a discriminatory and detrimental impact on millions of Americans participating in state exchanges that do not provide bariatric surgery coverage. We examined 3 state exchanges in which a portion of their plans provided coverage for bariatric surgery to determine if bariatric surgery coverage is correlated with premium costs. State health exchanges; United States. Data from the 2015 state exchange plans were analyzed using information from the Centers for Medicare & Medicaid Services' Individual Market Landscape file and Benefits and Cost Sharing public use files. Only 3 states (Oklahoma, Oregon, and Virginia) in the analysis have 1 or more rating regions in which a portion of the plans cover bariatric surgery. In Oklahoma and Oregon, the average monthly premiums for all bronze, silver, and gold coverage levels are higher for plans covering bariatric surgery. Only 1 of these states included platinum plans that cover bariatric surgery. The average difference in premiums was between $1 to $45 higher in Oklahoma, and $18 to $32 higher in Oregon. Conversely, in Virginia, the average monthly premiums are between $2 and $21 lower for each level for plans covering bariatric surgery. Monthly premiums for plans covering versus not covering bariatric surgery ranged from 6% lower to 15% higher in the same geographic rating region. Across all 3 states in the sample, the average monthly premiums do not differ consistently on the basis of whether the state exchange plans cover bariatric surgery.


Ant-nest soil and seedling growth in a neotropical ant-dispersed herb.
A major hypothesis concerning the benefits of myrmecochory, seed dispersal by ants, to plants is that ant nests are nutrient-enriched microsites that are beneficial to seedling growth. We experimentally test this hypothesis for a neotropical myrmecochore, Calathea ovandensis, asking two questions: 1) is soil of nests of a seed-dispersing ant chemically or structurally distinct from surrounding soils, and 2) do seedlings grow better in soil collected from ant nests than in randomly collected soil? We found that although ant-nest soil was significantly enriched in nitrate-nitrogen, magnesium, iron, manganese, cadmium and percent organic matter compared to randomly collected soil, seedling growth was not significantly improved by ant-nest soil.


Rapid protein identification using a microscale electrospray LC/MS system on an ion trap mass spectrometer.
A methodology has been developed for the rapid identification of gel separated proteins. Following in gel protein digestion with trypsin, the resulting peptide mixture is analyzed by on-line liquid chromatography, electrospray mass spectrometry (LC/MS). The mass spectral data containing either accurate mass values or sequence specific fragment ion information is then matched to a database of known protein sequences. Key features of the LC/MS system are the use of a novel integrated, microscale LC column-electrospray interface and variable flow solvent delivery to optimize the efficiency of sample loading and gradient elution. With these enhancements, only 10 min is required to analyze each sample. The method is routine for sample amounts ranging from 50 to 500 fmol. The analysis parameters for the ion trap mass spectrometer have to be carefully adjusted in order to keep pace with the rapidly eluting LC peaks. Although designed for rapid LC separations, the integrated column-electrospray interface is also able to provide extended analyses of selected components using a technique known as "peak parking".


The effect of liposome size on the final lipid/DNA ratio of cationic lipoplexes.
Several studies have demonstrated that lipoplexes are two-phase systems over most mixing lipid/DNA charge ratios. Because these studies have focused on small unilamellar vesicles (SUV), they leave open the question as to whether a similar pattern is followed by other liposome types. The main purpose of this work is to examine the question further by characterizing the assembly of cationic lipoplexes prepared from 1-[2-(oleoyloxy)ethyl]-2-oleyl-3-(2-hydroxyethyl)imidazolinium chloride (DOTIM)/dioleoylphosphatidylethanolamine (DOPE) (1:1) liposomes of various types. Sedimentation in sucrose density gradients reveals that large unilamellar vesicles (LUV) and sedimented multilamellar vesicles (sMLV), as opposed to SUV, form lipoplexes that exist as a single phase over a relatively broad range of mixing (+/-) ratios. This is indicated by observing that most of the LUV and sMLV become involved in the assembly reaction up to mixing (+/-) ratios of 4 and 9, respectively, while only a small and constant fraction of SUV associates with DNA at all mixing (+/-) ratios tested. Consequently, while maximal (+/-) ratios of approximately 4.5 and 9 are found in LUV and sMLV lipoplexes, respectively, a final (+/-) ratio of only approximately 2 is determined in SUV lipoplexes. Isothermal titration calorimetry shows that this is the lowest possible charge ratio achieved when liposomes are titrated with DNA. Based on these observations and on the size differences of the liposomes used, a model of lipoplex formation is proposed.


Lnc-PLA2G4A-4 facilitates the progression of hepatocellular carcinoma by inducing versican expression via sponging miR-23b-3p.
Aberrant expression of long non-coding RNAs (lncRNAs) is associated with progression of multiple human cancers including hepatocellular carcinoma (HCC). However, the role of lncRNAs in HCC is not been fully understood. Our study aimed to investigate the biological function and potential molecular mechanism of Lnc-PAL2G4A-4 in HCC. In the current study, we show that Lnc-PLA2G4A-4 was significantly up-regulated in HCC tissues and high Lnc-PLA2G4A-4 expression was remarkably associated with tumor size, microvascular invasion and poor prognosis of HCC patients. Functionally, Lnc-PLA2G4A-4 positively regulated cell proliferation, invasion and migration in vitro, and facilitated lung metastasis of HCC in vivo. Mechanistically, Lnc-PLA2G4A-4 functioned as a competing endogenous RNA (ceRNA) to bind to miR-23b-3p and subsequently facilitate miR-23b-3p's target gene versican (VCAN) expression in HCC cells. Over-expression of miR-23b-3p could reverse Lnc-PLA2G4A-4 induced cell phenotypes in HCC and suppress versican expression of by rescue analysis. Collectively, Lnc-PLA2G4A-4 promotes HCC progression by targeting the miR-23b-3p/versican axis, which may be a potential biomarker and therapeutic target for HCC.


Cascade Path Augmentation Unet for bladder cancer segmentation in MRI.
Treatment choices for patients with bladder cancer (BCa) are determined by the presence of muscular invasion. The precise segmentation of the inner and outer walls (IW and OW), as well as the bladder tumor (BT), is crucial for improving computer-aided diagnosis of muscle-invasive bladder cancer (MIBC). To propose a novel deep learning-based model to improve the segmentation accuracy of the IW, OW, and BT, which can be useful in clinical practice. We proposed a Cascade Path Augmentation Unet (CPA-Unet) network to conduct multi-regional segmentation of the bladder using 1545 T2-weighted MRI scans. The model employs a cascade strategy to eliminate the redundant information in the background. Unet is used to segment the bladder from the background in the rough segmentation. The path augmentation structure is used in the fine segmentation to mine multi-scale features. Additionally, the partial dense connection is adopted as the skip connection module to concatenate the low- and high-level sematic features. The CPA-Unet is trained using 1391 T2WI slices and tested using 154 T2WI slices. In comparison to previous deep learning-based methods, the CPA-Unet achieves superior segmentation results in terms of Dice similarity coefficient (DSC) and Hausdorff distance (HD) (IW: DSC = 98.19%, HD = 2.07 mm; OW: DSC = 82.24%, HD = 2.62 mm; BT: DSC = 87.40%, HD = 0.76 mm). Our proposed CPA-Unet network is capable of segmenting the bladder into its IW and OW, as well as tumors. The segmentation results provide a reliable and effective foundation for computer-assisted clinical diagnosis of MIBC.


Propagated Perturbations from a Peripheral Mutation Show Interactions Supporting WW Domain Thermostability.
Inter-residue interactions stabilize protein folds and facilitate allosteric communication. Predicting which interactions are crucial and understanding why remain challenging. We highlight this through studies of a single peripheral mutation (Q33E) on the surface of the Pin1 WW domain that causes an unexpected loss of thermostability. Nuclear magnetic resonance studies attribute the loss to reorganizations of electrostatic and hydrophobic interactions, resulting in propagated conformational perturbations. The propagation demonstrates the cooperative response of Pin1 WW to external perturbations, consistent with its allosteric behavior within Pin1. Microsecond molecular dynamics simulations suggest the wild-type fold relies on couplings between a surface electrostatic network and a highly conserved hydrophobic core; Q33E directly perturbs the former, thereby disrupting the latter. These couplings suggest that predictions of mutation consequences that assume dominance of a single interaction type can be limiting, and highlight challenges in predicting protein mutational landscapes.


Effect of calcitonin gene-related peptide on contraction of striated muscle in the mouse.
We have found ultrastructurally calcitonin gene-related peptide-like immunoreactivity in the axon terminal within the synaptic trough of neuromuscular junction of the mouse. We determined, using pharmacological means, with a phrenic nerve-diaphragm preparation, that this peptide enhances muscle contraction during stimulation of the nerve fibers or direct stimulation of the muscle. This effect is probably brought about via the receptor for this peptide not the acetylcholine receptor.


Neuronal involution during ageing. Ultrastructural study in the rat cerebellum.
Involutive phenomena have been investigated by electron microscopy in the Purkinje Pk neuron of the cerebellar cortex of the aging rat. The still limited number of specimens available to date, however, suggest an age-related progression of morphological and functional deteriorations involving particularly the intraneuronal "nucleus-ribosome system" (NRS). The impairments are characterized by changes in the nucleolar texture. These alterations are accompanied by modifications in the repartition and relative proportion of RNP components of the nucleolus. In addition, other nuclear elements such as interchromatin and perichromatin granules may vary in importance with age. Recognizable changes in the ribosomal constituents of the NRS are evidenced by modifications in the density and distribution of free ribosomes. An altered structure and organization of GER cisternae are also evident. Furthermore, "light" cytoplasmic areas, an increased evidence of neurotubules and the gradual congestion of the pericaryon by age pigments are other valuable ultrastructural features that may be regarded as part of the sequence of morphologic events occurring during neuonal ageing. The above ultrastructural data will subsequently form the basis of a model of ageing in the nerve cell, which will complete the previously proposed model of neuronal maturation. Therefore, this long-term study essentially purports the investigation of subcellular events taking place in the Pk neuron all along the normal life span in rats. This model will also be used to evaluate the changes in the sequence and the reinforcement of the processes of evolution versus involution as affected by certain xenobiotics, such as abused drugs(alcohol and narcotics). The intraneuronal modifications found in the nuclear and cytoplasmic structures of the NRS could possibly reflect the molecular dysfunction related to the production of various types of RNA and neuronal proteins. This hypothesis is supported by biochemical data obtained from analysis of the brain of aged animals. Ultrastructural and biochemical data appear to be in good agreement with the neurophysiologic interpretation of a slow-down and reduced efficiency of the CNS during the progressive development of senescence in human and animal subjects.