Bioactive constituents of Chinese natural medicines. VII. Inhibitors of degranulation in RBL-2H3 cells and absolute stereostructures of three new diarylheptanoid glycosides from the bark of Myrica rubra.
Three new diarylheptanoid glycosides, named (+)-S-myricanol 5-0-beta-D-glucopyranoside, myricanene A 5-O-alpha-L-arabinofuranosyl(1-->6)-beta-D-glucopyranoside, and myricanene B 5-0-alpha-L-arabinofuranosyl(1-->6)-beta-D-glucopyranoside, were isolated from the bark of Chinese Myrica rubra, together with twenty known compounds. The absolute stereostructures of the new diarylheptanoid glycosides were elucidated on the basis of chemical and physicochemical evidence, including the application of the modified Mosher's method. The inhibitory effects of isolated constituents on the release of beta-hexosaminidase from RBL-2H3 cells were examined, and several diarylheptanoids, myricanol, (+)-S-myricanol, myricanone, and myricanenes A and B, and a flavonol, myricetin, were found to show the inhibitory activity.


Association of p53 polymorphisms and colorectal cancer: modulation of risk and progression.
p53 Gene variants BstUI RFLP at codon 72 in exon 4, 16bp tandem repeat in intron 3 and MspI RFLP in intron 6, which code for two functionally different protein isoforms, have been shown to modulate susceptibility to different types of human neoplasms. p53 genotype was assessed in 90 CRC patients, 321 age-matched controls and 322 centenarians. The p53 codon 72 arginine, the p53 16bp deletion, and the MspI RFLP were significantly more frequent in CRC patients in comparison to the controls and to the centenarians (odd ratio 1.44 and 1.93). In the CRC group, the BstUI RFLP polymorphism was the more frequent combination (62.2%), and it was significantly associated with highly infiltrating (p<0.01), poorly differentiated (p<0.01), and metastatic (p<0.05) tumours. Our findings indicate that the p53 codon 72 polymorphisms are associated with a higher risk of CRC and are associated with more advanced and undifferentiated tumours.


New species of the Monocotylidae (Monogenea) from the stingray Dasyatis tortonesi Capapé (Euselachii, Dasyatidae) off the Tunisian coast, with comments on host-specificity and the specific identities of Mediterranean stingrays.
Heterocotyle capapei n. sp. and Thaumatocotyle tunisiensis n. sp. are described from the gills and the nasal cavity, respectively, of Dasyatis tortonesi collected from the Tunisian coast. H. capapei is distinguished from other species in the genus by the morphology of the penis, which is conical and has a long narrow distal point. T. tunisiensis differs from all known species of Thaumatocotyle by the shape and size of the penis. The host Dasyatis tortonesi has long been confused with D. pastinaca. Our study shows that their monogeneans are oioxenic, suggesting that D. pastinaca and D. tortonesi are distinct species. These results suggest that the monogenean parasite fauna could be used as a diagnostic tool for species identification: these parasites may, therefore, represent good biological indicators. The host-specificity of the Heterocotylinae and Merizocotylinae is discussed.


Fabrication and characterisation of high performance polypyrrole modified microarray sensor for ascorbic acid determination.
In this study, gold microelectrode array (Au/MEA) with electrode of 12 μm diameter was fabricated by photolithography technique. Subsequently, polypyrrole (Ppy) modified gold microarrays sensor (Ppy/Au/MEA) was prepared by cyclic voltammetry technique. The deposition potential range and number of cycles were optimised in order to get optimum thickness of Ppy film. Scanning Electron Microscope and Atomic Force Microscope investigations reveal that Ppy coating formed at 3 cycles is porous with thickness of 1.5 μm which exhibiting high catalytic current for ascorbic acid (AA) in square wave technique (SWV). In contrast to earlier sensors designs, these Ppy/Au/MEA sensors exhibits lower detection limit (LOD) of 10 nm towards AA at physiological conditions. It also exhibits enhanced sensitivity (2.5 mA cm(-2) mM(-1)) and long range of linear detection limit from 10 nm to 2.8 mM. In the same way, polypyrrole modified macro Au (Ppy/Au/MA) biosensor was also fabricated and its electro catalytic property towards AA was compared with that of Ppy/Au/MEA. The Ppy/Au/MA exhibits sensitivity of only 0.27 mA cm(-2) mM(-1), LOD of 5 μM and linear range of 10 μM to 2.2mM. Hence, our investigations indicate that the Ppy/Au/MEA could serve as highly sensitive sensor for AA than any of the earlier designs. So, the Ppy/Au/MEA electrode was utilised for determination AA in a wide variety of real samples.


Identification of Novel Variants in the PVRL1 Gene in Patients With Nonsyndromic Cleft Lip With or Without Cleft Palate.
Nonsyndromic cleft lip with/without cleft palate (nsCL/P, OMIM 119530) is one of the most common birth defects with a prevalence of ∼1/1000 in Caucasians. Studies have demonstrated an association between nsCL/P and the variants of the poliovirus receptor like-1 gene (PVRL1). The aim of this study was to describe novel variants in exon 3 of the PVRL1 gene and to investigate the association between exon 3 of the PVRL1 gene and Turkish patients with nsCL/P. 205 Turkish subjects were enrolled: 80 nsCL/P patients and 125 unrelated control individuals. Genomic DNA was isolated from peripheral blood leukocytes, and exon 3 of the PVRL1 gene was amplified using polymerase chain reaction (PCR). After PCR, the amplied DNA was sequenced using an automated sequencer. We identified two new variants of the PVRL1 gene at codons 174 and 187 in exon 3. These variants had nucleotide substitutions 520T>A and 560C>A, resulting in S174T and T187N amino acid changes, respectively. Two novel variants of the PVRL 1 gene were identified in nsCL/P patients. These findings suggest that PVRL1 variants make a contribution to nsCL/P in Turkish patients.


Revisiting and updating molecular epidemiology of α-thalassemia mutations in Thailand using MLPA and new multiplex gap-PCR for nine α-thalassemia deletion.
α-thalassemia is an inherited blood disorder that is most frequently found in Southeast Asian populations. In Thailand, molecular characterization can diagnose most patients with α-thalassemia; however, several atypical patients are also observed in routine analyses. Here, we characterized α-thalassemia mutations among 137 Hemoglobin H (Hb H) disease patients and three fetuses of Hb Bart's hydrops, a fatal clinical phenotype of α-thalassemia. Specifically, we performed multiplex ligation-dependent probe amplification (MLPA) followed by direct DNA sequencing. We noticed common genotypes in 129 patients and eight patients had rare Hb H disease caused by compound heterozygous α0-thalassemia (--CR or --SA deletion) with α+-thalassemia (-α3.7/-α4.2/αConstant Springα). Furthermore, two affected fetuses had the --SA/--SEA and one had the --CR/--SEA genotypes. Next, we developed and validated a new multiplex gap-PCR and applied this method to 844 subjects with microcytic red blood cells (RBCs) from various parts of Thailand. The frequency of heterozygous α0-thalassemia was dominated by --SEA 363/844 (43%), followed by --THAI 3/844 (0.4%), --SA 2/844 (0.2%), and --CR 2/844 (0.2%) mutations. These findings suggest that aforementioned four mutations should be routinely applied to increase the effectiveness of diagnosis and genetic counseling in this region.


The integration of the internal and external milieu in the insula during dynamic emotional experiences.
Whilst external events trigger emotional responses, interoception (the perception of internal physiological states) is fundamental to core emotional experience. By combining high resolution functional neuroimaging with concurrent physiological recordings, we investigated the neural mechanisms of interoceptive integration during free listening to an emotionally salient audio film. We found that cardiac activity, a key interoceptive signal, was robustly synchronised across participants and centrally represented in the posterior insula. Effective connectivity analysis revealed that the anterior insula, specifically tuned to the emotionally salient moments of the audio stream, serves as an integration hub of interoceptive processing: interoceptive states represented in the posterior insula are integrated with exteroceptive representations by the anterior insula to highlight these emotionally salient moments. Our study for the first time demonstrates the insular hierarchy for interoceptive processing during natural emotional experience. These findings provide an ecologically-valid framework for elucidating the neural underpinnings of emotional deficits in neuropsychiatric disorders.


The role of frailty on quality of life in older adults during the COVID-19 pandemic.
The impact of frailty, a physiological state characterized by reduced reserve for stressors and related to worse outcomes, on older adults during the pandemic is unclear. Our aim was to identify the effects of frailty among older adults during the COVID-19 pandemic. A total of 197 older adults who were not exposed to COVID-19 were assessed with an online survey one year after the pandemic began in Turkey. Frailty, quality of life, and fear of COVID-19 were assessed with the Tilburg Frailty Indicator, the Nottingham Health Profile, and the Fear of COVID-19 Scale, respectively. Since March 2020, changes in pain severity and localization, fatigue, and fear of falling were assessed. Multiple linear regression analyses were conducted. In this study, 62.5% of the participants were frail. The prevalence of pain was significantly increased during the COVID-19 pandemic, but only among the frail. The increases in pain severity, fear of falling, and fatigue were significantly higher for the frail than the non-frail. The model including physical and psychological components of frailty and pain severity explained 49% of the variation in quality of life (R = 0.696; R2 = 0.485; p < 0.001). The physical component of frailty had the highest impact on quality of life (B = 20.591; β = 0.334). This study focused on negative outcomes that were experienced more by frail older adults compared to non-frail older adults when they were locked down at home for an extended period of time during the COVID-19 pandemic. It is necessary to quickly improve and maintain the health of these affected individuals.


The effects of starving and refeeding on the intra-acinar distribution pattern of alcohol-dehydrogenase activity in rat liver.
Microquantitative determinations of ADH activity were carried out on the livers of male and female rats. The animals were either starved for 84 h, or starved and then refed with a carbohydrate-rich diet for 6 nights. When the enzyme activity is expressed in mumoles/min/g dry weight, fasting does not appear to alter liver ADH activity, while in starved and subsequently refed rats it is diminished by 20%. Microquantitative measurements of ADH activity in 50-150 ng lyophilized tissue samples, microdissected the whole way along the sinusoidal length, made the computer-aided plotting of intra-acinar distribution patterns possible. The results showed that, under the feeding conditions selected, only minor changes in the ADH activity profiles occur in the liver acinus. These are within the range of the standard deviations of the normal mean values. From these results it can be deduced that fasting and refeeding do not lead to specific inhibition or induction of liver ADH activity. - The decrease of ADH activity of total liver (mumol/min) per total body weight in starved rats is obviously the result of a loss of protein which affects the liver cells of all acinar zones almost equally.


Improving human rights in mental health takes more than just changing the law: An audit of medical assessments in regional community patients in Queensland.
To evaluate the medical assessments of involuntary community patients in a regional mental health service, determine the compliance with requirements under Queensland's Mental Health Act 2016 (the Act) to regularly review orders and assess patients' mental capacity. We audited 183 patient records on community treatment authorities (CTAs) to determine whether medical assessments undertaken under the Act included consideration of the person's capacity, and regular reviews by an authorised doctor as required1s205. The audit revealed that 51% of the CTA patients did not comply with legal requirements either to complete a capacity assessment and/or be medically assessed within three months of the last review. Over 50% of medical assessments did not comply with the legislative requirements to record capacity assessments and review involuntary treatment on at least a three-month basis. However, when the treatment criteria were met, it did not appear to be a basis for CTA revocation. Further research may help determine whether the Mental Health Review Tribunal (Tribunal) could play a greater role in overseeing compliance with the new legislative requirements or if other clinical oversight mechanisms would be appropriate to improve the assessment process.


The effect of lycopene on serum level of cardiac biomarkers in patients undergoing elective percutaneous coronary intervention: A randomized controlled clinical trial.
Myocardial ischemia may recur in a significant subset of patients following percutaneous coronary intervention (PCI). Lycopene, a carotenoid with antioxidant activity, has evidence for beneficial effects on cardiovascular system. In the present study, we aimed to evaluate the possible preventive effect of lycopene against post-PCI myocardial damage by detection of cardiac biomarkers of ischemia. A total of 45 patients who planned to undergo elective PCI were randomly assigned to two groups to receive either lycopene (30 mg 12 hours before PCI as well as 15 mg just before and 8 hours after PCI) along with standard treatment (n = 23) or only standard treatment (n = 22). Standard treatment included aspirin, a statin, and a beta-blocker. The serum levels of creatine kinase-MB (CK-MB), troponin I, and high sensitivity C-reactive protein (hs-CRP) were measured 12 hours before and 12 hours after the procedure and were compared between the two groups. The use of lycopene significantly prevented the increase of CK-MB following PCI compared to control (P = 0.048). However, it had not any significant effect on serum levels of troponin I (TnI) (P = 0.176) and hs-CRP (P = 0.186) compared to control. Lycopene can prevent the increase of CK-MB following PCI. Therefore, it has the potential for prevention of post-PCI cardiovascular events. However, more studies are needed to confirm such an effect.


Plate-and-bone-strut fixation of distal third humeral shaft aseptic non-unions: A consecutive case series.
Non-union after humeral shaft fractures are seen frequently in clinical practice. The incidence is 2-10% after conservative management and up to 30% after surgical treatment. The purpose of this study is to evaluate the outcomes of plate-and-bone-strut-allograft technique with bone chip augmentation for aseptic non-unions of the distal third of the humerus. 26 consecutive cases were treated using a trans-triceps approach. The non-union was fixed with a 4.5 mm Locking Compression Plate combined with a strut bone allograft at the anterior part of the humerus and bone chips. All patients underwent the same rehabilitation protocol of 12 weeks. Clinical evaluation took place 12 months after surgery with the Mayo elbow score and Oxford elbow score. Complete bone healing without complications was achieved in all 26 patients. The average period of radiographic union was 106 days. The average range of flexion-extension was 108° (94°-180°) and pro-supination was 159° (102°-180°). Twelve months after surgery, average Mayo elbow score was 86 (68-100) and the Oxford elbow score was 83 (52-100). The plate-and-bone-strut-allograft technique with bone chip augmentation in distal humeral shaft for aseptic non-unions resulted in union of all cases. No adverse events related to the surgery or the materials used were documented.


Author Correction: Abundance and diversity of the faecal resistome in slaughter pigs and broilers in nine European countries.
In the version of this Article originally published, the surname of author Oksana Lukjancenko was spelt incorrectly as 'Lukjacenko'. This has now been corrected.


Definitive pathology at radical prostatectomy is commonly favorable in men following initial active surveillance.
Limited data are currently available regarding the outcomes of radical prostatectomy (RP) in men with low-risk prostate cancer who were initially managed by active surveillance (AS). To evaluate the pathologic outcomes of patients who underwent RP following initial AS. We analyzed the records of 67 patients who underwent RP following initial AS begun between 1993 and 2011. All patients underwent confirmatory biopsy to reassess eligibility for AS. RP was recommended for disease progression suggested by follow-up biopsies or imaging. Unfavorable disease was defined as having at least one of the following pathologic findings: Gleason score (GS) ≥4+3, extracapsular extension of tumor, seminal vesicle invasion, or lymph node involvement. A descriptive analysis was performed to assess pathologic features. Median time from confirmatory biopsy to RP was 1.7 yr (range: 0.3-7.8). Reasons for discontinuing AS to undergo RP included evidence of increased tumor volume or grade on follow-up biopsy, patient preference/anxiety, and findings on follow-up imaging in 46 patients (68.7%), 17 patients (25.3%), and 4 patients (6.0%), respectively. Pathologic analyses revealed organ-confined disease in 55 patients (82.1%), and GS was ≥4+3 in 9 (13.4%). Positive nodes were observed in three patients (4.4%) and positive surgical margin in two (3.0%). Overall, 19 patients (28.4%) had unfavorable disease. Of the biopsy criteria for triggering RP, Gleason patterns >3 were the most frequently associated with unfavorable disease (43.3%). One patient (1.5%) experienced biochemical recurrence during postoperative follow-up (median: 3.2 yr). Our study may be limited by its retrospective and single-institution nature. Most patients who started initially on AS after undergoing confirmatory biopsy showed pathologically organ-confined disease with a low GS at RP. Such findings provide further evidence that, overall, AS is a safe treatment approach.


Teaching and training in old age psychiatry: a general survey of the World Psychiatric Association member societies.
One of the main goals of the World Psychiatric Association (WPA) section on old age psychiatry is to promote its discipline and consequently to help member societies to promote and develop locally specific medical education programmes in old age psychiatry. In order to target its support to the needs expressed by the member societies, the section conducted a survey within the WPA during the year 2001. A questionnaire was sent to each of the 116 WPA member societies in 93 countries. It assessed the level of development of medical education in old age psychiatry in each country, as well as the perceived needs in educational material. Answers were obtained from 50 WPA member societies (response rate 54%) from 48 countries (response rate 52%). The existence of specific old age psychiatry services was reported in 40 (83%) countries, but the discipline is recognized as a subspecialty in only 13 (27%). Formal teaching in old age psychiatry was reported at the undergraduate level in 44 (92%) countries. Specific learning objectives for postgraduate training were reported in 30 (86%) of the countries where the discipline is not yet recognized as a subspecialty. Specific Continuing Medical Education (CME) programs were however reported in only 50% of the countries. Support for the development of educational material was mostly thought to be useful at the postgraduate level. Depression, dementia, psychopharmacology and psychotherapy were among the most frequently cited areas in which educational material might be usefully provided. Support for the development of postgraduate training is seen as the most pressing need by WPA member societies. Such support could help to increase the level of recognition of the discipline locally and hence provide more effective support to older people with mental disorders.


Paroxysmal nocturnal hemoglobinuria and liver transplantation, a new paradigm.
Paroxysmal nocturnal hemoglobinuria (PNH) is a type of hemolytic anemia acquired by the PIG-A gene mutation. This causes a deficiency of a complement regulatory protein, CD59, which results in hemolysis, hemoglobinuria and thrombosis (due to the release of procoagulant factors). Budd-Chiari syndrome is characteristic in these patients and has classically been considered as a contraindication for liver transplantation (LT) due to post-transplant recurrence. Since the approval of eculizumab for the treatment of PHN, disease control is possible and therefore the post-transplant recurrence of thrombotic phenomena involving the liver is avoided.


Intact and total insulin-like growth factor-binding protein-3 (IGFBP-3) levels in relation to breast cancer risk factors: a cross-sectional study.
Levels of insulin-like growth factor (IGF)-I and its main binding protein (IGFBP-3) have been associated with breast cancer risk among premenopausal women. However, associations of IGFBP-3 levels with breast cancer risk have been inconsistent, possibly due to the different predominant forms of circulating IGFBP-3 (intact versus fragmented) that were measured in these studies. Here, we examine the association of breast cancer risk factors with intact and total IGFBP-3 levels. This cross-sectional study includes 737 premenopausal women recruited at screening mammography. Plasma intact and total IGFBP-3 and IGF-I levels were measured by enzyme-linked immunosorbent assay methods. Percent and absolute breast density were estimated using a computer-assisted method. The associations were evaluated using generalized linear models and Pearson (r) or Spearman (rs) partial correlation coefficients. Means +/- standard deviations of intact and total IGFBP-3 levels (ng/mL) were 1,044 +/- 234 and 4,806 +/- 910, respectively. Intact and total IGFBP-3 levels were correlated with age and smoking. Levels of intact IGFBP-3 were negatively correlated with waist-to-hip ratio (WHR) (r = -0.128; P = 0.0005), parity (rs = -0.078; P = 0.04), and alcohol intake (r = -0.137; P = 0.0002) and positively correlated with energy intake (r = 0.075; P = 0.04). In contrast, total IGFBP-3 levels were positively correlated with WHR (r = 0.115; P = 0.002), parity (rs = 0.089; P = 0.02), body mass index (BMI) (r = 0.115; P = 0.002), physical activity (r = 0.118; P = 0.002), and IGF-I levels (r = 0.588; P < 0.0001) and negatively correlated with percent or absolute breast density (r = -0.095; P = 0.01 and r = -0.075; P = 0.04, respectively). Our data show that associations of some breast cancer risk factors with intact levels of IGFBP-3 are different from those with total (intact and fragmented) IGFBP-3 levels. These findings suggest that different molecular forms of IGFBP-3 may bear different relations to premenopausal breast cancer risk.


Linalool: Monoterpene alcohol effectiveness in chronic synovitis through lowering Interleukin-17, spleen and thymus indices.
Interleukin-17 has a positive role in the initial induction and late chronic phases of many inflammatory disorders like arthritis. This cytokine has a strong option for therapeutic targeting due to the fact that it was found in the inflamed joints of individual with rheumatoid arthritis (RA) and persuasive evidence from experimental arthritis models indicating its pro-inflammatory actions. IL-17 suppression lessened the asperity of arthritis. The present study aimed to assess the anti-arthritic potential of linalool in a model of chronic joint inflammation (CFA-mediated rheumatoid arthritis) in rats. Linalool markedly lowered spleen and thymus indices as opposed to arthritic control. The over-formation of IL-17, COX-2, TNF-α IL-1β, iNOS and IL-6 were markedly impaired in all linalool treated rats, but IL-10 was raised as compared to arthritic animals in Real time-PCR. There was reduction in associated parameters like paw volume, arthritic index, mobility score, and flexion pain score and a marked increase in stance score in CFA model as compared to the arthritic control group. Furthermore, there was improvement in body weight, hematological, tissue, and radiological parameters in the CFA-model. Molecular docking study exhibited strong binding interaction of linalool with IL-17, PGE-2, iNOS and COX-2, thus providing a good correlation among experimental and theoretical results. The current findings show that linalool reduces adjuvant arthritis by suppressing pro-inflammatory mediators, arthritic development, and spleen and thymus indices. Thus, linalool may be employed therapeutically to alleviate arthritis in humans.


Geographical variations in the occurrence of leprosy: possible roles played by nutrition and some other environmental factors.
The considerable variation in leprosy prevalence among 35 villages and field areas in South India was compared with variations in their socioeconomic and nutritional conditions. Using villages/field areas and aggregates thereof as observational units, leprosy prevalence rates were found to be significantly lower in field areas than in villages. Moreover, there was a significant correlation between the occurrence of malnutrition in children 1 to 4 years of age and the prevalence of leprosy. The possible association between the occurrence of malnutrition in childhood and leprosy is discussed in light of the present knowledge of leprosy, immunocompetence, and malnutrition. Strategies for further illumination of the hypothesized importance of malnutrition among children are discussed.


The effects of the Promoting the Big and Quashing the Small Policy on pollutants from a coal power supply chain perspective.
China implemented the Promoting the Big and Quashing the Small Policy (PBQSP) in 2007 to alleviate air pollution due to coal-fired power plants (CFPPs). However, how the policy affected the air pollutant emissions remains poorly understood. We compared the air pollutant emissions between decommissioned and newly-built CFPPs under the PBQSP by developing a plant-level supply chain emission inventory model, taking Shandong, China's top provincial-level coal power producer, as the case area. The results indicated that compared to a 27.4% increase in electricity produced in Shandong from 2010 to 2014, the NOx, particulate matter (PM), Hg, and SO2 emissions disproportionately increased by 10.3%, 11.7%, 11.8% and 0.5%, respectively. It was found that the PBQSP policy has made great contribution to air pollution mitigation, as the supply chain emission intensities (emissions per kilowatt-hour) of NOX, PM, Hg and SO2 of the newly-built CFPPs are three-quarters, two-fifths, three-fifths, and four-fifths lower, respectively, than those of the decommissioned CFPPs. However, coal transport route changes caused an increase in NOX, PM, and SO2 emissions by 44.0%, 45.5%, and 55.6%, respectively, during the transportation stage, which offset the PBQSP overall mitigation effect. By building a comprehensive assessment framework for air pollutant control policies, our study provides insights for formulating coordinated mitigation measures for the pollutant-intensive industries, such as promoting efficient air pollutant control technologies including the spray dust suppression devices.


Targeting the right population for T3 + T4 combined therapy: where are we now and where to next?
The universal applicability of levothyroxine (LT4) monotherapy for the treatment of hypothyroidism has been questioned in recent years. Indeed, it is now clear that about 10-15% of LT4-treated hypothyroid patients are dissatisfied with their treatment. It is plausible that this subset of hypothyroid patients may need T3 + T4 combined therapy to restore peripheral euthyroidism. To address this issue, many clinical trials have investigated the effect of T3 + T4 combinations versus standard LT4-based therapy. However, to date, results have been inconclusive, mainly due to the lack of markers that identify candidates for combination therapy. A breakthrough in this field came with the recent finding that several single-nucleotide polymorphisms in the deiodinase genes are associated with the persistence of hypothyroid symptoms in biochemically euthyroid LT4-treated patients, and are thus markers of candidates for combination therapy. In addition, whole-genome association studies are expanding our knowledge of other genes of the thyroid hormone (TH) pathway that affect serum TH levels. To target the right population for the T3 + T4 combined therapy, the next step is to translate these new findings into prospective trials. Hopefully, this will pave the way to personalized therapy for each hypothyroid patient.


Prevalence and mechanisms of secondary intraocular pressure elevation in eyes with intraocular tumors.
A survey of 2704 eyes with intraocular tumors in patients who were evaluated on the Oncology Service at Wills Eye Hospital showed that 126 of the tumor-containing eyes (5%) had tumor-induced elevated intraocular pressure (IOP) at the time of diagnosis of the tumor. Of the 2111 eyes with uveal melanomas, secondary IOP elevation was present in 55 (3%). Secondary IOP elevation was present in 7% of eyes with iris melanoma, 17% with ciliary body melanoma, and 2% with choroidal melanoma. The most common mechanism of elevated IOP was tumor invasion of the angle in the case of iris melanomas, pigment dispersion and tumor invasion of the angle in the case of ciliary body melanomas, and iris neovascularization in the case of choroidal melanomas. Of the 256 eyes with uveal metastases, secondary IOP elevation was found in 12 eyes (5%). Secondary IOP elevation was present in 64% of eyes with iris metastases, 67% with ciliary body metastases, and 1% with choroidal metastases. The most common mechanism of elevated IOP was tumor invasion of the angle in the case of iris and ciliary body metastases, and angle closure in the case of choroidal metastases. There were 303 eyes with retinoblastoma, 17% of which had elevated IOP which was secondary to iris neovascularization in 70% of cases and to an angle closure without neovascularization in 27%. Several other intraocular tumors including lymphoma, leukemia, medulloepithelioma, melanocytoma, and adenoma of the iris pigment epithelium were occasionally associated with secondary elevated IOP.


A SAS macro for direct adjusted survival curves based on Aalen's additive model.
Aalen's additive model defines the covariate effects on the survival outcome to be time-varying. The aim of this paper is to explore the application of Aalen's model in assessing treatment effects, adjusting for possibly different patient profiles between the treatment groups. We treat Aalen's model as the underlying model and compute the direct adjusted survival curves for different treatments. The treatment comparison can be performed by finding a confidence interval for the difference between two direct adjusted survivals. We developed a SAS macro that calculates the direct adjusted survival estimates and the standard errors, based on Aalen's model. In this paper, we give two examples to explain the syntax for using the macro.


Natural catalyst for luminol chemiluminescence: application to validate peroxide levels in commercial hair dyes.
Here, we report a hydrothermally treated green leaves (Moringa oleifera) extract exploited as an efficient and highly sensitive catalyst to catalyze the chemiluminescence (CL) reaction of luminol. In the absence of enhancer, this green and hydrothermally treated catalyst was found to significantly enhance the CL intensity ~3.5-fold compared with the traditionally used K3 Fe(CN)6 catalyst. The structure and surface morphology of the catalyst was elucidated using X-ray photoelectron spectroscopy, scanning electron microscopy, X-ray diffraction, and Raman spectroscopy. The synergistic effect of the catalyst in the CL reaction was systematically investigated in the presence of hydrogen peroxide using ultraviolet-visible and CL spectroscopy. Studies showed that the sensitivity of the catalyst could be amplified by adjusting several parameters such as pH of the medium and concentrations of the base and luminol. The sensitivity of the novel-type catalyst was examined through the validation of hydrogen peroxide levels in commercial hair dye samples. Markedly, the catalyst displayed ultrasensitivity to hydrogen peroxide as the limit of detection of hydrogen peroxide using this catalyst was determined to be 0.02 μM under optimized conditions. In general, the proposed inexpensive, ecofriendly, and nontoxic catalyst could enable the determination of hydrogen peroxide for diverse analytical applications.


An evaluation study of reported pancreatic adenocarcinoma risk-associated SNPs from genome-wide association studies in Chinese population.
Previous genome-wide association studies (GWAS), and a pathway study of pancreatic ductal adenocarcinoma (PDAC) identified 14 significantly associated single nucleotide polymorphisms (SNPs) along with another 7 promising loci in European, Japanese, and Chinese descents. In this study, we aimed to evaluate the potential association of these SNPs with PDAC risk in the Chinese population. In this Chinese population-based case-control study with 254 cases and 1200 controls, we tested 20 PDAC risk associated SNPs from previous GWAS and one SNP from a pathway-based study. All 21 SNPs were polymorphic in the Chinese population. Twenty SNPs were included in the final analysis after the quality check (QC). Among these SNPs, three were significantly associated with PDAC risk after Bonferroni correction (P < 2.5E-03) including rs7779540 (at 7q36.2, P = 3.89E-06, OR = 2.59, 95%CI: 1.73-3.87), rs10919791 (at 1q32.1, P = 6.07E-05, OR = 1.52, 95%CI: 1.24-1.86) and rs401681 (at 5p15.33, P = 5.15E-04, OR = 1.42, 95%CI: 1.17-1.73). Rs2255280 (at 5p13.1, P = 8.16E-03, OR = 1.31, 95%CI: 1.07-1.6) showed significant association at the p < 0.05 level. The directions of effect of these SNPs were consistent with previous studies. Four PDAC risk-associated SNPs identified in GWAS of various populations are associated with PDAC risk in the Chinese population. Information on PDAC risk-associated SNPs and their ORs may facilitate risk assessment of PDAC risk in the Chinese population.


Comprehensive analysis of long non-coding RNA PVT1 gene interaction regulatory network in hepatocellular carcinoma using gene microarray and bioinformatics.
PVT1 has been reported to be involved in the tumorigenesis and development of different cancers. However, the role of PVT1 in hepatocellular carcinoma (HCC) remains unclear. In this study, we applied gene microarray analysis to detect differentially expressed genes (DEGs) between PVT1 RNAi groups and controls. We initially investigated and confirmed PVT1 expression in HCC using The Cancer Genome Atlas (TCGA). The potential genes and pathways associated with PVT1 were also analyzed. We also performed bioinformatics analyses (Gene Ontology (GO), pathway, Kyoto Encyclopedia of Genes and Genomes (KEGG), and network analyses) to explore the underlying pathways and networks of these potential genes. We selected DLC1 for further analysis. Based on the TCGA database, PVT1 was markedly up-regulated in HCC, whereas DLC1 was down-regulated. Moreover, PVT1 expression negatively correlated with DLC1 in HCC, an observation that has been further validated in different cohorts with Oncomine. High expression of PVT1 was positively associated with gender, race, vascular invasion and pathological grade in HCC. Additionally, the ROC curve indicated that both PVT1 and DLC1 have high diagnostic value in HCC. We speculated that PVT1 might play a significant role in HCC development and progression via regulation of various pathways and genes, especially DLC1 and the Hippo signaling pathway. However, this mechanism should be confirmed by functional experiments.


Astrocytes as gate-keepers in optic nerve regeneration--a mini-review.
Animals that develop without extra-embryonic membranes (anamniotes--fish, amphibians) have impressive regenerative capacity, even to the extent of replacing entire limbs. In contrast, animals that develop within extra-embryonic membranes (amniotes--reptiles, birds, mammals) have limited capacity for regeneration as adults, particularly in the central nervous system (CNS). Much is known about the process of nerve development in fish and mammals and about regeneration after lesions in the CNS in fish and mammals. Because the retina of the eye and optic nerve are functionally part of the brain and are accessible in fish, frogs, and mice, optic nerve lesion and regeneration (ONR) has been extensively used as a model system for study of CNS nerve regeneration. When the optic nerve of a mouse is severed, the axons leading into the brain degenerate. Initially, the cut end of the axons on the proximal, eye-side of the injury sprout neurites which begin to grow into the lesion. Simultaneously, astrocytes of the optic nerve become activated to initiate wound repair as a first step in reestablishing the structural integrity of the optic nerve. This activation appears to initiate a cascade of molecular signals resulting in apoptotic cell death of the retinal ganglion cells axons of which make up the neural component of the optic nerve; regeneration fails and the injury is permanent. Evidence specifically implicating astrocytes comes from studies showing selective poisoning of astrocytes at the optic nerve lesion, along with activation of a gene whose product blocks apoptosis in retinal ganglion cells, creates conditions favorable to neurites sprouting from the cut proximal stump, growing through the lesion and into the distal portion of the injured nerve, eventually reaching appropriate targets in the brain. In anamniotes, astrocytes ostensibly present no such obstacle since optic nerve regeneration occurs without intervention; however, no systematic study of glial involvement has been done. In fish, vigorously growing neurites sprout from the cut axons and within a few days begin to re-enervate the brain. This review offers a new perspective on the role of glia, particularly astrocytes, as "gate-keepers;" i.e., as being permissive or inhibitory, by comparison between fish and mammals of glial function during ONR.


Ca2+ binding, ATP-dependent Ca2+ transport, and total tissue Ca2+ in embryonic and adult avian dystrophic pectoralis.
Avian muscular dystrophy is an autosomal recessive genetic disease characterized by early hypertrophy and loss of function of pectoralis major. The disease is progressive, ultimately resulting in atrophy and heavy lipid deposition. Previous investigators have noted a decrease in the ability of the dystrophic sarcoplasmic reticulum to concentrate Ca2+. More recently, other investigators have shown an abnormal calcium uptake in avian dystrophic sarcoplasmic reticulum. They indicated, using freeze-fracture techniques, that a 90 A particle of the vesicle membrane exhibited a decreased population and suggested that they might be the ATPase involved in calcium transport. Our studies confirm earlier observations of a decreased rate of Ca2+ uptake and Ca2+ binding capacity of dystrophic fragmented sarcoplasmic reticulum vesicles which are isolated from both embryonic and adult pectoralis. These observations correlate in turn with a 75% drop in the Ca:ATP transport efficiency of the dystrophic sarcoplasmic reticulum determined by measuring the rate of 32Pi liberation from gamma-ATP32 during active calcium transport by the isolated sarcoplasmic reticulum SR. In addition, we have found a quantitative deficiency in a 65,000 dalton component of the dystrophic fragmented SR at the time of myoblast fusion by measuring 35S-Methionine incorporation into the SR, coupled to high resolution polyacrylamide gel electrophoresis and radioautography. Analysis of total tissue calcium by atomic absorption spectroscopy revealed a decrease in the total calcium content of dystrophic muscle.


Gemcitabine-retinoid prodrug loaded nanoparticles display in vitro antitumor efficacy towards drug-resilient human PANC-1 pancreatic cancer cells.
The treatment of pancreatic cancer with gemcitabine is hampered by its rapid metabolism in vivo, the dense stroma around the tumor site which prevents the drug from reaching the cancerous cells and drug resistance. To address these challenges, this study describes the preparation of a retinoid prodrug of gemcitabine, GemRA (gemcitabine conjugated to retinoic acid), and its formulation into a nanoparticulate system applicable for pancreatic cancer treatment. Retinoic acid targets stellate cells which are part of the stroma and can thus augment the delivery of gemcitabine. GemRA dissolved in dimethylsulfoxide presented efficacy towards PANC-1 (human) and mT4 (mouse) pancreatic cancer cell lines but its poor solubility in aqueous solution affects its applicability. Thus, the preparation of the nanoparticles was initially attempted through self-assembly of GemRA, which resulted in the formation of unstable aggregates that precipitated during preparation. As a result, encapsulation of the drug into micelles of polyethylene glycol-retinoic acid (PGRA) amphiphilic conjugates was accomplished and resulted in successful incorporation of GemRA into nanoparticles of ca. 33 nm by dynamic light scattering and 25 nm by transmission electron microscopy. The nanoparticles had good stability in aqueous media and protected gemcitabine from the enzymatic action of cytidine deaminase, which converts gemcitabine to its inactive metabolite upon circulation. Cellular uptake of the nanoparticles by PANC-1 cells was confirmed by fluorescence spectroscopy and flow cytometry. Treatment of PANC-1 cells in vitro with the prodrug-loaded nanoparticles resulted in a significant reduction in cell viability (IC50 ca. 5 μM) compared to treatment with gemcitabine (IC50 > 1000 μM). The ability of the GemRA-loaded nanoparticles to induce cellular apoptosis of treated PANC-1 cells was ascertained via a TUNEL assay suggesting these nanoparticles are effective in pancreatic cancer treatment.


Deep investigation on different effects of Cl- in transformation of reactive species in Fe(II)/NH2OH/PDS and Fe(II)/NH2OH/H2O2 systems.
Hydroxylamine (NH2OH) has been verified to efficiently strengthen pollutants oxidation in Fe(II)/peroxydisulfate (PDS) and Fe(II)/H2O2 systems. However, the different effects of hydroxylamine salts types were rarely recognized. Herein, the effects of two commonly used hydroxylamine salts (i.e. NH2OH·HCl and (NH2OH)2·H2SO4) on oxidation kinetics and reactive species composition were compared in Fe(II)/PDS and Fe(II)/H2O2 systems for the first time. Pseudo first order kinetics could only describe benzoic acid (BA) oxidation well in Fe(II)/NH2OH/H2O2 system, which was related to the different concentration changes of Fe(III) determined by [Formula: see text] . Hydroxylamine salts types influenced not kinetic rules, but reaction rates of target compounds. The empirical reaction rate constant of BA in Fe(II)/NH2OH·HCl/PDS system was 141.5% of that in Fe(II)/(NH2OH)2·H2SO4/PDS system under the same concentration of NH2OH (1.4 mM), while the apparent reaction rate constant in Fe(II)/NH2OH·HCl/H2O2 system was 68% of that in Fe(II)/(NH2OH)2·H2SO4/H2O2 system. This opposite effect resulted from the differences in primary reactive species compositions and their interactions with Cl-. Reactive species identification indicated that Cl- would decrease the contribution of ferryl ion (Fe(IV)) and transform sulfate radical (SO4·-) to hydroxyl radical (·OH) in Fe(II)/NH2OH/PDS system, while it competitively consumed the only reactive species ·OH in Fe(II)/NH2OH/H2O2 system. This study highlights the importance of reductants types on strengthening Fenton oxidation and offers a reference for reasonable construction of the relevant systems.


Retrograde adsorption isotherms: an impossible fact?
"Retrograde" adsorption isotherms have been reported, but seem to violate thermodynamics and would cause concentration steps to migrate against the direction of fluid-phase flow. In general, what appears to be retrograde behavior is caused by one or more additional, uncontrolled variables. This is illustrated with adsorption of sulfonate on a weak-acid ion exchanger, where adsorption is accompanied by partial conversion of the ion exchanger to the sodium form.


Reorganization of mossy fiber synapses in male and female hypothyroid rats: a stereological study.
Thyroid hormone deficiency has been reported to interfere with synaptogenesis, particularly in those regions of the brain where the neurons display a late and protracted histogenesis, although the extent of the synaptic alterations remains unknown. To provide detailed quantitative data on the effects of hypothyroidism upon synapses, a link of the hippocampal circuitry was selected: the contact between mossy fibers and dendritic excrescences of CA3 pyramidal cells (MF-CA3 synapses). Groups of six male and six female rats aged 30 and 180 days were analyzed separately after being treated as follows: (1) hypothyroid from day 0 until day 30 (30 day old hypothyroid group); (2) hypothyroid from day 0 until day 180 (180 day old hypothyroid group); (3) hypothyroid until day 30 and thenceforth maintained euthyroid (recovery group); and (4) and (5) 30 and 180 day old control groups, respectively. Timm staining, Golgi impregnation, and electron microscopy were employed to estimate the volume of the mossy fiber system, the number and size of mossy fiber boutons, and the number and related features of MF-CA3 synapses. The volume of the mossy fiber system and the number of synaptic boutons were reduced in all experimental groups. The total number of synapses was decreased in 30 day old hypothyroid rats, but did not differ among 180 day old animals. Postsynaptic densities were shorter in hypothyroid and recovery groups than in controls, although the reduction was not as marked in recovery rats as it was in hypothyroid animals. Structural alterations were noted in the pre- and postsynaptic compartments of MF-CA3 synapses of both 180 day old hypothyroid and recovery rats. These changes can be regarded as mechanisms of reorganization as they underlie the compensation for the hypothyroid-induced numerical reduction of synapses observed in 30 day old animals and enable a complete catch-up of their total number. However, synaptic reorganization was not fully achieved, as revealed by the reduction in the size of the synaptic sites in hypothyroid and recovery animals. Finally, we demonstrate that hypothyroidism did not interfere with the sex-related differences of MF-CA3 synapses described in normal rats.


Ultrasound and colour Doppler in infantile subglottic haemangioma.
Subglottic haemangioma causes progressive and life-threatening stridor, typically manifesting at age 2-3 months. Standard diagnosis is by laryngoscopy. Larynx sonography is rarely used but allows assessment of the presence and extension of a mass that impinges on the subglottic airway. The additional use of colour Doppler enables demonstration of the vascular nature of such masses. To compare US and endoscopic findings in infants with subglottic haemangioma and to evaluate accuracy of US and colour Doppler imaging in this diagnosis. We report eight infants with subglottic haemangioma seen in our institution over the last decade. They presented with laryngeal stridor and were all investigated with both US and endoscopy. Six infants underwent colour Doppler sonography. US and endoscopic findings showed excellent anatomical correlation in lateral subglottic haemangioma. Colour Doppler imaging was deemed helpful in four infants. Larynx sonography with complementary colour Doppler imaging was non-invasive and helpful in the diagnosis of subglottic haemangioma.


Resistance to anti-HER2 therapy associated with the TSC2 nonsynonymous variant c.4349 C > G (p.Pro1450Arg) is reversed by CDK4/6 inhibitor in HER2-positive breast cancer.
HER2-positive breast cancer patients carrying the germline TSC2 nonsynonymous variant c.4349 C > G (p.Pro1450Arg) are resistant to anti-HER2 therapy. Multi-predictor in silico analysis reveals that this variant is deleterious. We explore the potential mechanism of this TSC2 variant and investigate methods for overcoming anti-HER2 resistance. TSC2 c.4349 C > G reverses the inhibitory effect on mTOR and downstream signaling by increasing TSC2 phosphorylation at Thr1462 and confers significant lapatinib resistance in vitro and in vivo. The combination of lapatinib and the CDK4/6 inhibitor palbociclib inhibits cyclin D1/CDK4/Rb alternative pathway and TSC2 phosphorylation, thereby partially attenuating mTOR activity and inducing TSC2-mutant cell blockage at G1/G0. In in vitro and xenograft models, palbociclib+lapatinib shows higher anti-tumor activity than monotherapy and overcomes the resistance of the TSC2 c.4349 C > G-related variant to anti-HER2 therapy. We reveal a new mechanism of resistance to anti-HER2 therapy and provide a strategy to increase the efficiency of anti-HER2 therapy in HER2-positive breast cancer.


Synthesis of the core of actinophyllic acid using a transannular acyl radical cyclization.
A synthetic study of actinophyllic acid based on an original strategy has been described. A transannular acyl radical cyclization allowed us to obtain a key bicyclo[3.3.2] framework, and construction of a core of the target alkaloid has been accomplished by subsequent introduction of a C2 unit.


An open-label trial of enhanced brief interpersonal psychotherapy in depressed mothers whose children are receiving psychiatric treatment.
Major depression affects one out of five women during her lifetime. Depressed mothers with psychiatrically ill children represent an especially vulnerable population. Challenged by the demands of caring for ill children, these mothers often put their own needs last; consequently, their depressions remain untreated. This population is especially difficult to engage in treatment. We have developed a nine-session intervention, an engagement session followed by eight sessions of brief interpersonal psychotherapy designed to increase maternal participation in their own psychotherapy, resolve symptoms of maternal depression, and enhance relationships (IPT-MOMS). This open-label trial assesses the feasibility and acceptability of providing this treatment to depressed mothers. Thirteen mothers meeting DSM-IV criteria for major depression were recruited from a pediatric mental health clinic where their school-age children were receiving psychiatric treatment. Subjects (mothers) were treated openly with IPT-MOMS. Eighty-five percent (11/13) completed the study. Subjects were evaluated with the Hamilton Rating Scale for Depression, and completed self-report measures of quality of life and functioning at three time points: baseline, after treatment completion, and 6-months posttreatment. A signed rank test was used to compare measurement changes between assessment time points. Subjects showed significant improvement from baseline to posttreatment on measures of maternal symptoms and functioning. These gains were maintained at 6-month follow-up. Therapy was well tolerated and accepted by depressed mothers, who are typically difficult to engage in treatment. A high proportion of subjects completed treatment and experienced improvements in functioning. Future randomized clinical trials are needed to establish the efficacy of this approach.


Computational analysis of the effects of the hERG channel opener NS1643 in a human ventricular cell model.
Dysfunction or pharmacologic inhibition of repolarizing cardiac ionic currents can lead to fatal arrhythmias. The hERG potassium channel underlies the repolarizing current I(Kr), and mutations therein can produce both long and short QT syndromes (LQT2 and SQT1). We previously reported on the diphenylurea compound NS1643, which acts on hERG channels in two distinct ways: by increasing overall conductance and by shifting the inactivation curve in the depolarized direction. The purpose of this study was to determine which of the two components contributes more to the antiarrhythmic effects of NS1643 under normokalemic and hypokalemic conditions. The study consisted of mathematical simulation of action potentials in a human ventricular ionic cell model in single cell and string of 100 cells. Regardless of external potassium concentration or diastolic interval used, NS1643 decreases action potential duration and triangulation. For single cells, NS1643 increases the postrepolarization refractory time but shortens the absolute refractory period. In one dimensional simulations, NS1643 increases the vulnerable window for unidirectional block but suppresses the emergence of premature action potentials and unidirectional blocks around APD(90). During normokalemia, shifting the inactivation curve has greater impact than increasing conductance, whereas the opposite occurs during hypokalemia. Increased hERG conductance and the depolarizing shift of the inactivation curve both contribute to the antiarrhythmic actions of NS1643, with relative effects dependent on external K(+) concentration.


Triose phosphate utilization in leaves is modulated by whole-plant sink-source ratios and nitrogen budgets in rice.
Triose phosphate utilization (TPU) is a biochemical process indicating carbon sink-source (im)balance within leaves. When TPU limits leaf photosynthesis, photorespiration-associated amino acid exports probably provide an additional carbon outlet and increase leaf CO2 uptake. However, whether TPU is modulated by whole-plant sink-source relations and nitrogen (N) budgets remains unclear. We address this question by model analyses of gas-exchange data measured on leaves at three growth stages of rice plants grown at two N levels. Sink-source ratio was manipulated by panicle pruning, by using yellower-leaf variant genotypes, and by measuring photosynthesis on adaxial and abaxial leaf sides. Across all these treatments, higher leaf N content resulted in the occurrence of TPU limitation at lower intercellular CO2 concentrations. Photorespiration-associated amino acid export was greater in high-N leaves, but was smaller in yellower-leaf genotypes, panicle-pruned plants, and for abaxial measurement. The feedback inhibition of panicle pruning on rates of TPU was not always observed, presumably because panicle pruning blocked N remobilization from leaves to grains and the increased leaf N content masked feedback inhibition. The leaf-level TPU limitation was thus modulated by whole-plant sink-source relations and N budgets during rice grain filling, suggesting a close link between within-leaf and whole-plant sink limitations.


Peroxyl radicals: inductors of neurodegenerative and other inflammatory diseases. Their origin and how they transform cholesterol, phospholipids, plasmalogens, polyunsaturated fatty acids, sugars, and proteins into deleterious products.
The most oxygen-sensitive constituents of cells are polyunsaturated fatty acids (PUFAs), which are incorporated in the outermost layer of cells in the form of phospholipids. PUFAs easily suffer oxidation. Identical marker compounds of these lipid peroxidation (LPO) processes are generated in both neurodegenerative and cardiovascular diseases, indicating a close relationship between the inducers of these events. Apparently, any alteration of the cell membrane structure influences the channels crossing the cell wall and causes an influx of Ca2+ ions. Ca2+ ions induce activation of phospholipases, which cleave phospholipids. Thus, the generated free PUFAs serve as substrates of lipoxygenases (LOXs) and cyclooxygenases. LOXs transform PUFAs into lipid hydroperoxides (LOOHs). If an outside impact exceeds a certain limit, the catalyzing bivalent iron ions in LOXs are liberated. They cleave the enzymatically generated LOOH molecules and induce a switch to nonenzymatic LPO reactions that produce peroxyl radicals (LOO*). Although LOO* radicals are also intermediates in enzymatic LPO processes, they are prevented from leaving the enzyme complex before the reaction is completed by generation of LOOH molecules. LOO* radicals are much more reactive than LOOH molecules and attack nearly all types of biological molecules. The generated products seem to serve as ligands for proteins that in turn induce gene activation. Thus, PUFA-phospholipids are apparently the precursor molecules of signal molecules that respond in a dose-related manner to any event that influences the cell structure by inducing an appropriate gene response. In this paper an overview of the deleterious chemical reactions initiated by LOO* radicals is presented. Many of these reactions have not been taken into account in previous research. These include epoxidation of cholesterol-PUFA esters, plasmalogens, and sphingolipids, as well as the release of hydrogen peroxide by the reaction of LOO* radicals with alcohols (sugars) and amines. The oxidation of proteins generating plaque formation involves only the LOO* radical-sensitive functional groups in side chains of the protein backbone and is therefore a rather late event in the development of Alzheimer disease and atherosclerosis.


Comparative analysis of chemical, mineral and in-vitro antibacterial activity of different varieties of date fruits from Saudi Arabia.
Three important varieties of dates (Sukri, Barhi, and Rothana), grown in Saudi Arabia were analyzed for their chemical, mineral and anti-bacterial activity. Among all the varieties analyzed, the Sukri dates had the highest moisture (16%), carbohydrate (80.65 mg/g) and the phenolic content (1.24 mg GAE/100 g) whereas, the ash content was the lowest (2.22%). The mineral content analyzed by ICP OES reveals that all three varieties contain an appreciable amount of essential macro and micro nutrients. They contain a good amount of potassium; the maximum amount of K (8171.24 mg/kg) was recorded in Barhi dates. Besides that, all three varieties of dates contain a good amount of Se, Cu, P, Fe, Mn. The comparative analysis of the GC MS chromatogram reveals that there were only five compounds that were present in all three varieties. The bioactive compounds Mannoic and Lactone were detected in all three varieties. The fruits contain some important groups of sugar, alcoholic sugar, aromatic hydrocarbon, phenol(s) and esters. The extracts of dates were assed for their antibacterial activity against 6 important strains of human pathogenic bacteria. The strains of E. coli-ATCC25922, B. subtilis-ATCC6633, S. aureus-ATCC25923, were observed to be highly sensitive to most of the extracts whereas S. pyogenes and S. flexeneri (clinical isolates) showed moderate sensitivity to some extracts. The variety Sukri was found to be superior to the other two varieties in terms of sugar and phenolic contents, in addition, the methanol extract of Sukri dates showed excellent antibacterial activity against most of the bacteria evaluated. These varieties of dates prove to have immense potential to be utilized as a therapeutic agent for curing mineral deficiency also can be explored for developing antibacterial agents.


Determination of azide in gastric fluid and urine by flow-injection electrospray ionization tandem mass spectrometry.
A rapid method was developed to identify and quantify the azide ion (N(3)(-)) in gastric fluid and urine. N(3)(-) in diluted biological fluids was reacted with NaAuCl(4) to produce Au(N(3))(2)(-), which was extracted with octanol. Five microliters of the extract were flow-injected into an electrospray ionization tandem mass spectrometric instrument. Quantification of N(3)(-) was performed by selected reaction monitoring of the product ion Au(N)(N(3))(-) at m/z 253, which was derived from the precursor ion Au(N(3))(2)(-) at m/z 281, using 50 μL of aqueous solution within 10 min. This method was found to be linear up to 10(-5) M, to have a limit of quantification of 10(-7) M, a limit of detection of 3.0 × 10(-8) M, and a coefficient of variation of ≦10% at 10(-7) M. In the case of urine, 50 μL of urine were spiked with N(3)(-), this was diluted 10-fold and passed through 1 mL of a resin, and finally diluted to 100-fold of the original. This method was linear up to 10(-3) M, had a limit of quantification of 10(-5) M, a limit of detection of 3.0 × 10(-6) M, and coefficient of variation of ≦8.8% for an original urine concentration of 10(-5) M. The practical applicability of this method was checked by diluting 1 μL of a suspected suicide victim's gastric fluid 20,000-fold and 1 μL of the victim's urine 5,000-fold and then measuring the N(3)(-) levels. These levels were found to be (7.5 ± 1.0) × 10(-2) M and (3.2 ± 0.4) × 10(-3) M, respectively.


A mutation in the catalytic loop of Hsp90 specifically impairs ATPase stimulation by Aha1p, but not Hch1p.
Heat shock protein 90 (Hsp90) is a molecular chaperone that plays a central role in maintaining cellular homeostasis by facilitating activation of a large number of client proteins. ATP-dependent client activation by Hsp90 is tightly regulated by a host of co-chaperone proteins that control progression through the activation cycle. ATPase stimulation of Hsp90 by Aha1p requires a conserved RKxK motif that interacts with the catalytic loop of Hsp90. In this study, we explore the role of this RKxK motif in the biological and biochemical properties of Hch1p. We found that this motif is required for Hch1p-mediated ATPase stimulation in vitro, but mutations that block stimulation do not impair the action of Hch1p in vivo. This suggests that the biological function of Hch1p is not directly linked to ATPase stimulation. Moreover, a mutation in the catalytic loop of Hsp90 specifically impairs ATPase stimulation by Aha1p but not by Hch1p. Our work here suggests that both Hch1p and Aha1p regulate Hsp90 function through interaction with the catalytic loop but do so in different ways.


Diagnostic value of tachycardia features and pacing maneuvers during paroxysmal supraventricular tachycardia.
The purpose of this prospective study was to quantitate the diagnostic value of several tachycardia features and pacing maneuvers in patients with paroxysmal supraventricular tachycardia (PSVT) in the electrophysiology laboratory. No study has prospectively compared the value of multiple diagnostic tools in a large group of patients with PSVT. One hundred ninety-six consecutive patients who had 200 inducible sustained PSVTs during an electrophysiology procedure were included. The diagnostic values of four baseline electrophysiologic parameters, nine tachycardia features and five diagnostic pacing maneuvers were quantified. The only tachycardia characteristic that was diagnostic of atrioventricular (AV) nodal reentry was a septal ventriculoatrial (VA) time of <70 ms, and no pacing maneuver was diagnostic for AV nodal reentry. An increase in the VA interval with the development of a bundle branch block was the only tachycardia characteristic that was diagnostic for orthodromic tachycardia, but it occurred in only 7% of all tachycardias. An atrial-atrial-ventricular response upon cessation of ventricular overdrive pacing was diagnostic of atrial tachycardia, and this maneuver could be applied to 78% of all tachycardias. Burst ventricular pacing excluded atrial tachycardia when the tachycardia terminated without depolarization of the atrium, but the result could be obtained only in 27% of patients. This prospective study quantitates the diagnostic value of multiple observations and pacing maneuvers that are commonly used during PSVT in the electrophysiology laboratory. The findings demonstrate that diagnostic techniques rarely provide a diagnosis when used individually. Therefore, careful observations and multiple pacing maneuvers are often required for an accurate diagnosis during PSVT. The results of this study provide a useful reference with which new diagnostic techniques can be compared.


Effects of cocaine self-administration on ethanol, food and water intake in the rat.
Because cocaine and ethanol are frequently abused in combination, the present study was performed to assess the behavioral consequences of concurrent access to both of these drugs. Rats trained to respond for food under a fixed-ratio 40 (FR 40) schedule of reinforcement during four, 30-min periods each day (every 6 h) were given free access to a 5% (w/v) ethanol solution and to water. Once a stable baseline of food, ethanol and water intake was established, intravenous cocaine was made available under a fixed-ratio 1 (FR1) schedule. After cocaine self-administration had been established for 10 days, the ethanol was removed from the chambers for 3 days. After the ethanol was returned to the chambers, saline was substituted for cocaine for 3 days. Following saline substitution, animals were given increased cocaine availability. Before cocaine was made available, rats consumed primarily the ethanol solution, averaging 31.7 ml of the 5% solution (4.2 g/kg ethanol), 10.2 ml of water and 148 food pellets/day. When cocaine (0.2 mg/kg per injection, i.v.) was made available, rats self-administered an average of 40-85 injections per day. Self-administration of cocaine resulted in small decreases in ethanol and food intake, as well as some changes in the pattern of ethanol intake. However, removing the ethanol from the chambers had no effect upon food and cocaine intake. Substitution of saline for the cocaine altered the pattern, but not the amount of ethanol intake. There was a trend toward increased ethanol intake during the study, which may have been related to repeated cycles of cocaine availability.


Butyrate prevents Harvey Sarcoma virus focus formation but permits oncogene expression.
Since butyrate inhibits DNA synthesis and cell growth, we adapted NIH 3T3 cells to grow in butyrate to study its effects on retroviral cell transformation. In cultures of butyrate-adapted NIH 3T3 cells infected with Harvey Sarcoma Virus, no foci were formed; however, there was evidence of retroviral replication, and the P21 oncogene product was demonstrated by immunofluorescence in its characteristic localization beneath the cytoplasmic membrane of single cells and pairs of cells. When the butyrate was removed, these cells proliferated to form foci. Butyrate therefore does not prevent oncogene expression but does reversibly inhibit focus formation.


Intestinal and multivisceral autotransplantation for tumors of the root of the mesentery: Long-term follow-up.
To present the indications, techniques, short- and long-term outcomes after visceral exenteration, ex vivo resection, and intestinal/multivisceral autotransplantation. All patients who have undergone this procedure at our center were studied. Technique, postoperative complications, survival, tumor recurrence, and functional status were recorded. Ten patients, 4 children and 6 adults, have undergone these procedures since January 1999. Seven patients are alive at 13-138 months later, 6 with functioning autografts and one after rescue with an allotransplantation. Intestinal/multivisceral autotransplantation is a potentially valuable option for some otherwise unresectable neoplasms of the root of the mesentery.


Bedside determination of microcirculatory oxygen delivery and uptake: a prospective observational clinical study for proof of principle.
Assessment of microcirculatory functional capacity is considered to be of prime importance for therapy guidance and outcome prediction in critically ill intensive care patients. Here, we show determination of skin microcirculatory oxygen delivery and consumption rates to be a feasible approach at the patient's bedside. Real time laser-doppler flowmetry (LDF) and white light spectrophotometry (WLS) were used for assessment of thenar skin microperfusion, regional Hb and postcapillary venous oxygen saturation before and after forearm ischemia. Adapted Fick's principle equations allowed for calculation of microcirculatory oxygen delivery and uptake. Patient groups with expected different microcirculatory status were compared [control (n = 20), sepsis-1/2 definition criteria identified SIRS (n = 10) and septic shock patients (n = 20), and the latter group further classified according to sepsis-3 definition criteria in sepsis (n = 10) and septic shock (n = 10)], respectively. In otherwise healthy controls, microcirculatory oxygen delivery and uptake approximately doubled after ischemia with maximum values (mDO2max and mVO2max) significantly lower in SIRS or septic patient groups, respectively. Scatter plots of mVO2max and mDO2max values defined a region of unphysiological low values not observed in control but in critically ill patients with the percentage of dots within this region being highest in septic shock patients. LDF and WLS combined with vasoocclusive testing reveals significant differences in microcirculatory oxygen delivery and uptake capacity between control and critically ill patients. As a clinically feasible technique for bedside determination of microcirculatory oxygen delivery and uptake, LDF and WLS combined with vasoocclusive testing holds promise for monitoring of disease progression and/or guidance of therapy at the microcirculatory level to be tested in further clinical trials.ClinicalTrials.gov: NCT01530932.


Proactive motor control within and between hands: Effects of age, motor set, and cue type.
Several studies have reported that proactive motor control in a cued four-finger choice reaction task proceeds more efficiently with a 2-hands motor set (two fingers on each hand) than with a 1-hand motor set (four fingers on one hand). According to the Grouping Model, this is because the 2-hands motor set recruits distinct left and right hand representations located in separate cerebral hemispheres, whereas the 1-hand motor set recruits partially overlapping neural areas grouped together in one hemisphere. The latter neural organization increases neuromotor noise, thereby complicating proactive motor selection. The present study examined the effect of older age on the 2-hands motor selection advantage. A group of young and a group of older adults performed two proactive motor tasks-the procue task and the anticue task-with two motor sets: a 2-hands and 1-hand set. Predictive cues preceded the target signal at five different time intervals (100-850 ms), allowing advance selection of 2 out of 4 fingers. Older adults showed longer reaction times and smaller cueing benefits compared to younger adults. Overall, cueing benefits were greater, and accrued faster, with the 2-hands than with the 1-hand motor set, reflecting the beneficial impact of the neuroanatomical hand distinction. Importantly, the 2-hands advantage was substantially greater in the older age group, suggesting that the hand distinction might abate age-related neural dedifferentiation. These findings highlight the impact of cortical representational distinctiveness in proactive motor control, especially in older age.


Clinically Important Differences for Mobility Measures Derived from the Testosterone Trials.
Accurate estimates of clinically important difference (CID) are required for interpreting the clinical importance of treatments to improve physical function, but CID estimates vary in different disease populations. We determined the CID for two common measures of walking ability in mobility-limited older men. Longitudinal, multisite placebo-controlled trial. Men enrolled in the Testosterone Trials who had self-reported mobility limitation and gait speed less than 1.2 m/second (n = 429). Testosterone- and placebo-allocated participants were combined for this study. Mean changes from baseline, adjusting for time-in-intervention and site, were 29.6, 13.2, 12.5, -2.4, and -32.6 m for 6MWD, and 15.4, 7.2, 2.1, -3.4, and -7.2 for PF10 in men who reported their mobility was "very/much better," "little better," "no change," "little worse," or "much worse," respectively. CID estimates using regression, ROC, and eCDF varied from 5.0-29.6 m for 6MWD, and 5.0-15.2 points for PF10. CID estimates vary by the population studied and by the method and precision of measurement. Increases of 16 to 30 m for 6MWD and 5 to 15 points for PF10 over 12 months appear to be clinically meaningful in mobility-limited, older hypogonadal men. These CID estimates may be useful in the design of efficacy trials of therapies to improve physical function.


Alcohol and multiple trauma: is there an influence on the outcome?
A relevant number of trauma patients are intoxicated with alcohol at admission in trauma centers. Meanwhile, some studies provide data suggesting a profound influence of ethanol on the posttraumatic clinical course; others could not confirm these findings. Knowledge of the influence of ethanol in a multiple trauma cohort is lacking. Therefore, we performed a retrospective outcome study of initially intoxicated multiple trauma patients in a German level-1 trauma center. Patients with an Injury Severity Score greater than or equal to 16 and aged 16-65 years were included in our study. Ventilation time, duration of intensive care unit treatment, the course of cytokines, and the incidence of systemic inflammatory response syndrome (SIRS), sepsis, and multiple organ dysfunction syndrome (MODS) were analyzed. Total in-patient time, mortality, and the requirement for blood products were evaluated. Logistic regression analyses were performed. Injury severity was comparable in both groups but there were more severe abdominal injuries in alcohol-intoxicated patients. The clinical course was comparable in both groups. Alcohol consumption was not an independent risk factor to sustain SIRS (odds ratio [OR], 1.00; 95% confidence interval [CI], 0.59-1.70), sepsis (OR, 0.84; 95% CI, 0.54-1.31), or for mortality (OR, 1.08; 95% CI, 0.53-2.13). There was a trend toward an increased incidence of MODS in alcohol-intoxicated patients (OR, 2.74; 95% CI, 0.90-8.35). Blood alcohol level at the time of admission is not a valuable marker for worse or improved outcome in multiple trauma patients. There were no ethanol-related differences concerning overall injury severity; however, more severe abdominal injuries were found in alcohol-intoxicated patients. There was no increased risk for posttraumatic complications in primarily alcohol-intoxicated multiple trauma patients.


Piezosurgery in the cochleostomy through multiple middle ear approaches.
Piezosurgery is a new instrument which is able to cut the bone without necrosis and non-mineralized tissues damage. The aim of this work has been to test Piezosurgery as a new and alternative method in cochleostomy. We have performed Piezosurgery on nine temporal bone specimens from voluntary bone donors. Piezosurgery was used in the mastoidectomy with posterior tympanotomy approach (three specimens), suprameatal approach (three specimens) and combined approach (three specimens). The piezoelectric device uses low frequency ultrasonic waves (24.7-29.5 kHz), the applied power can be modulated between 2.8 and 16 W, and is programmed in accordance to the density of the bone. The equipment consists of two handpieces, two inserts and two peristaltic pumps: the microvibrations that are created in the piezoelectric handpiece cause the inserts to vibrate linearly between 60 and 210 microm. In all the specimens, Piezosurgery provided excellent control without side effects on the adjacent structures of the middle and inner ear. Piezosurgery is a new and revolutionary osteotomy technique utilizing the microvibrations of scalpels at ultrasonic frequency, so that soft tissue will not be damaged even upon accidental contact with the cutting tip. The vibration frequency of Piezosurgery is optimal for mineralized tissue and does not cut the adjacent soft tissue, minimizing the risk of harming the adjacent tissues: this renders the piezoelectric device ideal for cochlear implantation in children with ossified, partial ossified cochlea and/or malformation of ear.


Structure of Biocompatible Coatings Produced from Hydroxyapatite Nanoparticles by Detonation Spraying.
Detonation-produced hydroxyapatite coatings were studied by scanning electron microscopy (SEM), X-ray powder diffraction (XRD), Raman spectroscopy, and electron paramagnetic resonance (EPR) spectroscopy. The source material for detonation spraying was a B-type carbonated hydroxyapatite powder. The coatings consisted of tetracalcium phosphate and apatite. The ratio depended slightly on the degree of crystallinity of the initial powder and processing parameters of the coating preparation. The tetracalcium phosphate phase was homogeneous; the apatite phase contained defects localized on the sixfold axis and consisted of hydroxyapatite and oxyapatite. Technological factors contributing to the transformation of hydroxyapatite powder structure during coating formation by detonation spraying are discussed.


Introduction for the special issue on "Tissue Barriers in Inflammation".
This issue of Tissue Barriers contains the inaugural special issue devoted to recent advances in barrier function of endothelial and epithelial cells. We used this opportunity to invite experts in vascular endothelial cell biology and epithelial cell biology to comment on critical questions and problems in permeability of organ and tissue barriers, and to provide insight into common areas in these fields, namely how these cells maintain homeostasis and response to injury and infection. To complement these reviews, this issue also contains four research articles that explore specific questions related respiratory and intestinal epithelial cell function.


Gelatine thin films as biomimetic surfaces for silica particles formation.
The formation of silica nanostructures by several living organisms, such as diatoms or sponges, involves specific macromolecules that control the growth and the organization of silica nanoparticles. In order to investigate if a single molecular system could perform both particle size control and morphological template, gelatine thin films of various concentration and strength were prepared as biomimetic models and their reactivity towards sodium silicate aqueous solutions was studied. Simultaneous formation of silica particles in the nanometric and micrometric size range was observed. The former corresponds to colloids grown at the surface of the gelatine films and the latter to particles induced by gelatine chain brushes formed at the film/water interface. These results are in good agreement with well-known principles of biomineralization and suggest that multi-molecular systems, rather than single components, are responsible for biogenic silica nanostructure formation.


Oral versus topical Ibuprofen for chronic knee pain: a prospective randomized pilot study.
Topical ibuprofen provides an alternative treatment to oral ibuprofen for the treatment of chronic knee pain. To compare the efficacy of topical versus oral ibuprofen in chronic knee pain treatment. Prospective, randomized, unblinded pilot study. A private pain management practice. Twenty patients received either ibuprofen tablets 3 times daily (2400 mg total) or 4% topical gel 4 times daily (320 mg total) for 2 weeks. Subjects completed the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index, the Medical Outcomes Study 12-Item Short Form (SF-12v2) Health Survey, and a satisfaction questionnaire. Comparison of WOMAC and SF-12v2 mean changes from baseline showed no differences between groups. Patient satisfaction and study treatment convenience were rated equivalently between groups. Within the topical group, significant improvements (P < 0.05) were experienced in the mean differences of WOMAC Pain scores from baseline to 2 weeks (-82.6, -158.3 to -6.8), WOMAC Stiffness scores from baseline to one week (-25.3, -50.0 to -0.6) and baseline to 2 weeks (-47.8, -95.7 to 0.1), WOMAC Physical Function scores from baseline to one week (-175.9, -348.6 to -3.2) and baseline to 2 weeks (-312.1, -580.5 to -43.7), and patient satisfaction scores from baseline to one week and baseline to 2 weeks. Within the oral group, significant improvements (P < 0.05) were experienced in mean differences of WOMAC Physical Function from baseline to one week (-342.6, -638.1 to -47.1) and baseline to 2 weeks (-323.2, -637.1 to -9.2). As this was a preliminary investigation, the sample size of 20 subjects is a limitation in this study. Treatment of chronic knee pain with topical ibuprofen provided comparable clinical efficacy and patient satisfaction as oral ibuprofen in this pilot study.


Diverse myocardial extension and autonomic innervation on ligament of Marshall in humans.
The ligament of Marshall (LOM) or the oblique vein of Marshall (VOM) in the left atrium (LA) is one of the origins of nonpulmonary vein ectopies causing atrial fibrillation. The distributions of myocardial bundles (Marshall bundles; MBs) and autonomic nerves adjacent to the LOM have not been completely elucidated. Twenty-eight human hearts were examined postmortem. The inferolateral LA myocardium was dissected en block and cut perpendicularly to the atrioventricular ring. The specimens were sectioned at 1-mm intervals and stained with hematoxylin and eosin, and Azan-Mallory. In addition, S-100 and tyrosine hydroxylase (TH) were employed for the immunohistochemical analysis of the sympathetic and parasympathetic fibers. In 25 cases, MBs were observed. Although the MBs surrounded the VOM at the coronary sinus (CS) juncture, they gradually diminished in density toward the distal venous branch. The connections of the MBs and LA myocardium were frequently observed in the CS juncture and anterior wall of the left pulmonary vein (PV)-LA junctions. TH-stained sympathetic nerve fibers were densely distributed around the PV-LA junctions. On the other hand, non-TH-stained parasympathetic ganglions were mainly observed at the CS juncture. MB-LA connections were mainly observed at the CS juncture and PV-LA junctions. The association between sympathetic nerve fibers and MBs was distinct at the PV-LA junctions. At the CS juncture, the regression of the sympathetic nerve fibers and the increase of parasympathetic ganglions were observed. The diverse arrangement and autonomic innervation of MBs seem to characterize the LOM region in humans.


Molecular dynamics study of lipid bilayers modeling the plasma membranes of normal murine thymocytes and leukemic GRSL cells.
Molecular dynamics (MD) calculations for the plasma membranes of normal murine thymocytes and thymus-derived leukemic GRSL cells in water have been performed under physiological isothermal-isobaric conditions (310.15K and 1 atm) to investigate changes in membrane properties induced by canceration. The model membranes used in our calculations for normal and leukemic thymocytes comprised 23 and 25 kinds of lipids, respectively, including phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, lysophospholipids, and cholesterol. The mole fractions of the lipids adopted here were based on previously published experimental values. Our calculations clearly showed that the membrane area was increased in leukemic cells, and that the isothermal area compressibility of the leukemic plasma membranes was double that of normal cells. The calculated membranes of leukemic cells were thus considerably bulkier and softer in the lateral direction compared with those of normal cells. The tilt angle of the cholesterol and the conformation of the phospholipid fatty acid tails both showed a lower level of order in leukemic cell membranes compared with normal cell membranes. The lateral radial distribution function of the lipids also showed a more disordered structure in leukemic cell membranes than in normal cell membranes. These observations all show that, for the present thymocytes, the lateral structure of the membrane is considerably disordered by canceration. Furthermore, the calculated lateral self-diffusion coefficient of the lipid molecules in leukemic cell membranes was almost double that in normal cell membranes. The calculated rotational and wobbling autocorrelation functions also indicated that the molecular motion of the lipids was enhanced in leukemic cell membranes. Thus, here we have demonstrated that the membranes of thymocyte leukemic cells are more disordered and more fluid than normal cell membranes.


Problem-solving and decision-making preferences: no difference between complementary and alternative medicine users and non-users.
The objective of this study was to determine if complementary and alternative medicine (CAM) Users are more autonomous than Non-Users with respect to their preferred role in decision-making (measured by the problem-solving decision-making or PSDM scale). A survey was mailed in spring 2001 to a random sample of 696 men (selected from the Ontario Cancer Registry), aged 18 years or older and diagnosed with prostate cancer in the preceding 2 years. Less than 5% of the 489 (72.1% response rate) men (mean age 68.6 years) who responded to the PSDM question in our survey were classified as having an autonomous role preference, while almost 1/3 of the respondents reported using CAM for their prostate cancer. The majority of respondents were classified as preferring a shared role and a substantial minority was classified as preferring a passive role. There was no statistically significant difference between CAM users and non-users with respect to their preferred role. The hypothesis that CAM Users are more autonomous problem solvers and decision makers is not supported by these findings; however, the generalizability of our results is limited by the fact that we surveyed a relatively older male population only.


Recombinant murine interferon-gamma inhibits the fusion of mouse alveolar macrophages in vitro but stimulates the formation of osteoclastlike cells on implanted syngeneic bone particles in mice in vivo.
Osteoclasts are multinucleated cells that originate from the fusion of mononuclear precursors and are responsible for bone resorption. Indirect evidence from in vitro studies suggests that IFN-gamma and TNF-alpha inhibit and stimulate bone resorption, respectively, but contradictory results have emerged from the literature regarding the effects of IFN-gamma on macrophage multinucleation. Using highly sensitive model systems, the present work demonstrates that, in mice, rMuIFN-gamma inhibits the fusion of alveolar macrophages in vitro but augments the number of osteoclastlike cells on implanted syngeneic bone particles in vivo. Although rMuTNF-alpha fails to stimulate macrophage multinucleation in either system, treatment of implanted animals with rMuIFN-gamma appears to limit the inflammatory reaction and favor tissue repair.


High-throughput analysis of concentration-dependent antibody self-association.
Monoclonal antibodies are typically monomeric and nonviscous at low concentrations, yet they display highly variable associative and viscous behavior at elevated concentrations. Although measurements of antibody self-association are critical for understanding this complex behavior, traditional biophysical methods are not capable of characterizing such concentration-dependent self-association in a high-throughput manner. Here we describe a nanoparticle-based method, termed self-interaction nanoparticle spectroscopy, that is capable of rapidly measuring concentration-dependent self-interactions for three human monoclonal antibodies with unique solution behaviors. We demonstrate that gold nanoparticles conjugated with antibodies at low protein concentrations (<40 μg/mL) display self-association behavior (as measured by the interparticle distance-dependent plasmon wavelength) that is well correlated with static light-scattering measurements obtained at three orders of magnitude higher antibody concentrations. Using this methodology, we find that the antibodies display a complex pH-dependent self-association behavior that is strongly influenced by the solution ionic strength. Importantly, we find that a polyclonal human antibody is nonassociative for all solution conditions evaluated in this work, suggesting that antibody self-association is more specific than previously realized. We expect that our findings will guide rational manipulation of antibody phase behavior, and enable studies that elucidate sequence and structural determinants of antibody self-association.


Morphological Changes of Human Corneal Endothelial Cells after Rho-Associated Kinase Inhibitor Eye Drop (Ripasudil) Administration: A Prospective Open-Label Clinical Study.
To investigate the effect and safety of a selective Rho kinase inhibitor, ripasudil 0.4% eye drops, on corneal endothelial cells of healthy subjects. Prospective, interventional case series. In this study, 6 healthy subjects were administered ripasudil 0.4% in the right eye twice daily for 1 week. Morphological changes and corneal endothelial cell density were examined by noncontact and contact specular microscopy. Central corneal thickness and corneal volume of 5 mm-diameter area of center cornea were analyzed by Pentacam Scheimpflug topography. All the above measurements were conducted in both eyes before administration, 1.5 and 6 hours after the initial administration on day 0; and in the same manner after the final administration on day 7. By noncontact specular microscopy, indistinct cell borders with pseudo guttae were observed, but by contact specular microscopy, morphological changes of corneal endothelial cells were mild and pseudo guttae was not observed after single and repeated administration of ripasudil in all subjects. These changes resolved prior to the next administration, and corneal endothelial cell density, central corneal thickness and corneal volume were not changed throughout the study period. Transient morphological changes of corneal endothelial cells such as indistinct cell borders with pseudo guttae were observed by noncontact specular microscopy in healthy subjects after ripasudil administration. Corneal edema was not observed and corneal endothelial cell density did not decrease after 1 week repetitive administration. These morphological changes were reversible and corneal endothelial cell morphology returned to normal prior to the next administration. JAPIC Clinical Trials Information 142705.


An ontogenic study of amygdala seizures induced by penicillin in rats.
Amygdaloid seizures were induced by unilateral injection of penicillin into the amygdala in infant and young rats to investigate the ontogenic development of the amygdaloid seizure. "Repetitive isolated discharges" and "sustained high-frequency discharges" appeared in 3- to 4-day-old rats. In these rats, the repetitive discharges appeared in bilateral subcortical structures simultaneously with those of the amygdaloid penicillin focus, but sustained high-frequency discharges were localized to the ipsilateral subcortical structures. In 9- to 10-day-old and 14- to 15-day-old rats, repetitive discharges occurred simultaneously in the bilateral cortices and subcortical structures. In contrast to them, sustained high-frequency discharges propagated faster from the amygdala penicillin focus to the bilateral midbrain reticular formation and medial nucleus of the thalamus than to other regions. In 20- to 21-day-old and older rats, both events occurred simultaneously at all cerebral sites. These results indicate that the subcortical structures, especially the midbrain reticular formation and medial nucleus of the thalamus, play an important role in seizure propagation from the ipsilateral to the contralateral hemisphere in infant rats.


Uncovering the emergence of HSCs in the human fetal bone marrow by single-cell RNA-seq analysis.
During fetal development, human hematopoietic stem cells (HSCs) colonize the bone marrow (BM), where they self-renew and sustain hematopoiesis throughout life; however, the precise timepoint at which HSCs seed the BM is unclear. We used single-cell RNA-sequencing to map the transcriptomic landscape of human fetal BM and spleen hematopoietic stem/progenitor cells (HSPCs) and their microenvironment from 10 to 14 post-conception weeks (PCWs). We further demonstrated that functional HSCs capable of reconstituting long-term multi-lineage hematopoiesis in adult NOG mice do not emerge in the BM until 12 PCWs. In contrast, functional HSCs were not detected in the spleen by 14 PCWs. By comparing the niche-HSPC interactions between BM and spleen, we identified ligand-receptor pairs likely to be involved in fetal HSC migration and maintenance. Our work paves the way for research into the mechanisms underlying HSC colonization in human fetal BM and provides invaluable resources for future studies on HSC development.


Prescribing of neuraminidase inhibitors for influenza in UK primary care since the 2009 pandemic.
We determined prescribing rates of neuraminidase inhibitors (NIs) for influenza in UK primary care since 2009 in relation to national prescribing guidelines. All NI prescriptions issued during the influenza seasons between October 2010 and May 2013 were extracted from The Health Improvement Network (THIN), a large UK primary-care database. We calculated NI prescribing rates per 100,000 person-weeks (pw) by age group, sex, deprivation level, influenza season and presence of chronic conditions with 95% confidence intervals (CIs), and used negative binomial regression models to determine the independent association between these variables and NI prescribing. NI prescribing was rare. The prescribing rate was 1·7/100,000 pw (95% CI 1·7-1·8) during influenza-active periods, and 0·1/100,000 (95% CI 0·1-0·1) during non-active periods. Prescribing rates were highest in 25- to 44-year-olds in 2010/2011 and in persons aged ⩾85 years in 2011/2012 and 2012/2013. Individuals with chronic conditions had significantly higher prescribing rates than persons without (rate ratio 2·62, 95% CI 2·27-3·03). GPs are more likely to prescribe NIs to high-risk individuals and during influenza active periods, as per national guidelines. We could not assess the proportion of patients with influenza-like illness who were prescribed an NI.


Identification of miR-106b-93 as a negative regulator of brown adipocyte differentiation.
microRNAs (miRNAs) have been reported to play an essential role in the regulation of brown adipocyte adipogenesis. In the present study, we investigated the role of the miR-106b-93 cluster in the differentiation of brown adipocytes. We found that knockdown of miR-106b and miR-93 significantly induced the expression of brown fat-specific genes and promoted the accumulation of lipid-droplet in differentiating brown adipocytes. In addition, ectopic expression of miR-106b and miR-93 suppressed the mRNA level of Ucp1, a selective hallmark of brown adipocytes. Furthermore, the expression levels of miR-106b and miR-93 are higher in brown adipose tissues of high fat diet-induced obese mice compared to control mice. Taken together, our results identify miR-106b and miR-93 as negative regulators of brown adipocyte differentiation and the miR-106b-93 cluster may play an important role in regulating energy homeostasis.


Metal switch-controlled myosin II from Dictyostelium discoideum supports closure of nucleotide pocket during ATP binding coupled to detachment from actin filaments.
G-proteins, kinesins, and myosins are hydrolases that utilize a common protein fold and divalent metal cofactor (typically Mg(2+)) to coordinate purine nucleotide hydrolysis. The nucleoside triphosphorylase activities of these enzymes are activated through allosteric communication between the nucleotide-binding site and the activator/effector/polymer interface to convert the free energy of nucleotide hydrolysis into molecular switching (G-proteins) or force generation (kinesins and myosin). We have investigated the ATPase mechanisms of wild-type and the S237C mutant of non-muscle myosin II motor from Dictyostelium discoideum. The S237C substitution occurs in the conserved metal-interacting switch-1, and we show that this substitution modulates the actomyosin interaction based on the divalent metal present in solution. Surprisingly, S237C shows rapid basal steady-state Mg(2+)- or Mn(2+)-ATPase kinetics, but upon binding actin, its MgATPase is inhibited. This actin inhibition is relieved by Mn(2+), providing a direct and experimentally reversible linkage of switch-1 and the actin-binding cleft through the swapping of divalent metals in the reaction. Using pyrenyl-labeled F-actin, we demonstrate that acto·S237C undergoes slow and weak MgATP binding, which limits the rate of steady-state catalysis. Mn(2+) rescues this effect to near wild-type activity. 2'(3')-O-(N-Methylanthraniloyl)-ADP release experiments show the need for switch-1 interaction with the metal cofactor for tight ADP binding. Our results are consistent with strong reciprocal coupling of nucleoside triphosphate and F-actin binding and provide additional evidence for the allosteric communication pathway between the nucleotide-binding site and the filament-binding region.


Primary hyperparathyroidism due to ectopic parathyroid adenoma in an adolescent: a case report and review of the literature.
Primary hyperparathyroidism (PHPT) is a common endocrine disorder and is usually diagnosed in adults. PHPT due to ectopic parathyroid adenoma in adolescents is rare. We describe the case of a 15-year-old boy with PHPT due to ectopic parathyroid adenoma. A review of the literature of PHPT in adolescents was performed, focusing on etiology, clinical presentation, preoperative localization methods, pathology, and treatment. The patient was successfully treated with surgery and was followed up for 5 years with no signs or symptoms of hyperparathyroidism. By reviewing the literature, only seven cases of PHPT associated with ectopic parathyroid lesions in adolescents have been reported. Parathyroidectomy is the only known curative treatment. Accurate preoperative localization of the target lesion is critical. This study should raise awareness of the diagnosis and treatment of PHPT due to ectopic parathyroid adenoma/carcinoma in adolescents.