The Chicken Thigh Adductor Profundus Free Muscle Flap: A Novel Validated Non-Living Microsurgery Simulation Training Model.
Simulation training is becoming an increasingly important component of skills acquisition in surgical specialties, including Plastic Surgery. Non-living simulation models have an established place in Plastic Surgical microsurgery training, and support the principles of replacement, reduction and refinement of animal use. A more sophisticated version of the basic chicken thigh microsurgery model has been developed to include dissection of a type 1-muscle flap and is described and validated here. A step-by-step dissection guide on how to perform the chicken thigh adductor profundus free muscle flap is demonstrated. Forty trainees performed the novel simulation muscle flap on the last day of a 5-day microsurgery course. Pre- and post-course microvascular anastomosis assessment, along with micro dissection and end product (anastomosis lapse index) assessment, demonstrated skills acquisition. The average time to dissect the flap by novice trainees was 82±24 minutes, by core trainees 90±24 minutes, and by higher trainees 64±21 minutes (P=0.013). There was a statistically significant difference in the time to complete the anastomosis between the three levels of training (P=0.001) and there was a significant decrease in the time taken to perform the anastomosis following course completion (P<0.001). Anastomosis lapse index scores improved for all cohorts with post-test average anastomosis lapse index score of 3±1.4 (P<0.001). The novel chicken thigh adductor profundus free muscle flap model demonstrates face and construct validity for the introduction of the principles of free tissue transfer. The low cost, constant, and reproducible anatomy makes this simulation model a recommended addition to any microsurgical training curriculum.


Chromium effects on glucose tolerance and insulin sensitivity in persons at risk for diabetes mellitus.
To investigate the effects of daily chromium picolinate supplementation on serum measures of glucose tolerance and insulin sensitivity in patients at high risk for type 2 diabetes mellitus. We conducted a randomized, double-blind, placebo-controlled, modified cross-over clinical trial with 6-month sequences of intervention and placebo followed by a 6-month postintervention assessment. Adult patients with impaired fasting glucose, impaired glucose tolerance, or metabolic syndrome were enrolled. Participants received 6-month sequences of chromium picolinate or placebo at 1 of 2 dosages (500 or 1000 mcg daily). Primary outcome measures were change in fasting plasma glucose, 2-hour plasma glucose during oral glucose tolerance testing, fasting and 2-hour insulin, and homeostasis model assessment of insulin resistance (HOMA-IR). Secondary outcomes included anthropometric measures, blood pressure, endothelial function, hemoglobin A1c, lipids, and urinary microalbumin. Fifty-nine participants were enrolled. No changes were seen in glucose level, insulin level, or HOMA-IR (all P>.05) after 6 months of chromium at either dosage level (500 mcg or 1000 mcg daily) when compared with placebo. None of the secondary outcomes improved with either chromium dosage compared with placebo (P>.05). Chromium supplementation does not appear to ameliorate insulin resistance or impaired glucose metabolism in patients at risk for type 2 diabetes and thus is unlikely to attenuate diabetes risk.


Antiallergic activity of rosmarinic acid esters is modulated by hydrophobicity, and bulkiness of alkyl side chain.
Methyl, propyl and hexyl esters of rosmarinic, caffeic and p-coumaric acids were tested for antiallergic activity, and rosmarinic acid propyl ester exhibited the greatest β-hexosaminidase release suppression (IC50, 23.7 μM). Quadratic correlations between pIC50 and cLogP (r(2) = 0.94, 0.98, and 1.00, respectively) were observed in each acid ester series. The antiallergic activity is modulated by hydrophobicity, and alkyl chain bulkiness.


Transperitoneal Laparoscopic Pyelolithotomy versus Percutaneous Nephrolithotomy for Treating the Patients with Staghorn Kidney Stones: A Randomized Clinical Trial.
To compare the performance and outcomes of laparoscopic pyelolithotomy (LPL) versus percutaneous nephrolithotomy (PCNL) in the management of staghorn kidney stones. This study was a parallel-group randomized clinical trial study carried out on 68 patients with staghorn stones (one single piece or maximally two-piece stones with large extra renal part) over 18 years referred to Labbafinejhad Hospital. Patients were randomly divided on a ratio of 1:1 into two groups of LPL and PCNL using random allocation software. The primary outcome was the stone free rate, which was evaluated with KUB, and ultrasonography. Secondary outcomes were duration of surgery, bleeding, fever, post-operative pain, length of hospital stay, and postoperative complications. The mean±SD age of patients in PCNL and LPL groups were 48.50 ± 13.33 years and 52.17 ± 15.74 years, respectively (P=.303). LPL was associated with a higher duration of surgery (196.55 ± 26.58 minutes versus 110.88 ± 34.82; P=.001). Hemoglobin drop in the PCNL group was higher than the LPL group (2.67 ± 2.61 g/dL versus -0.7912 ± 1.06 g/dL; P=.001). Stone free status was observed in 29 (85.3%) patients in the LPL group, which was significantly higher than the PCNL group (22 patients, 64.7%; P =.050). The results of this study indicate that LPL offers a higher stone free rate with less bleeding in patients with single particle or limited particles staghorn stones with extrarenal pelvis but is associated with a higher duration of operation. The application of LPL in patients with multiple stones carries a lower achievement and is not encouraged.


Histone acetyltransferase (HAT) activity of p300 modulates human T lymphotropic virus type 1 p30II-mediated repression of LTR transcriptional activity.
Human T-lymphotropic virus type-1 (HTLV-1) is a deltaretrovirus that causes adult T cell leukemia/lymphoma, and is implicated in a variety of lymphocyte-mediated inflammatory disorders. HTLV-1 provirus has regulatory and accessory genes in four pX open reading frames. HTLV-1 pX ORF-II encodes two proteins, p13II and p30II, which are incompletely defined in virus replication or pathogenesis. We have demonstrated that pX ORF-II mutations block virus replication in vivo and that ORF-II encoded p30II, a nuclear-localizing protein that binds with CREB-binding protein (CBP)/p300, represses CREB and Tax responsive element (TRE)-mediated transcription. Herein, we have identified p30II motifs important for p300 binding and in regulating TRE-mediated transcription in the absence and presence of HTLV-1 provirus. Within amino acids 100-179 of p30II, a region important for repression of LTR-mediated transcription, we identified a single lysine residue at amino acid 106 (K3) that significantly modulates the ability of p30II to repress TRE-mediated transcription. Exogenous p300, in a dose-responsive manner, reverses p30II-dependent repression of TRE-mediated transcription, in the absence or presence of the provirus, In contrast to wild type p300, p300 HAT mutants (defective in histone acetyltransferase activity) only partially rescued p30(II)-mediated LTR repression. Deacetylation by histone deacetylase-1 (HDAC-1) enhanced p30II-mediated LTR repression, while inhibition of deacetylation by trichostatin A decreases p30(II)-mediated LTR repression. Collectively, our data indicate that HTLV-1 p30II modulates viral gene expression in a cooperative manner with p300-mediated acetylation.


A lysin motif effector subverts chitin-triggered immunity to facilitate arbuscular mycorrhizal symbiosis.
Arbuscular mycorrhizal (AM) fungi greatly improve mineral uptake by host plants in nutrient-depleted soil and can intracellularly colonize root cortex cells in the vast majority of higher plants. However, AM fungi possess common fungal cell wall components such as chitin that can be recognized by plant chitin receptors to trigger immune responses, raising the question as to how AM fungi effectively evade chitin-triggered immune responses during symbiosis. In this study, we characterize a secreted lysin motif (LysM) effector identified from the model AM fungal species Rhizophagus irregularis, called RiSLM. RiSLM is one of the highest expressed effector proteins in intraradical mycelium during the symbiosis. In vitro binding assays show that RiSLM binds chitin-oligosaccharides and can protect fungal cell walls from chitinases. Moreover, RiSLM efficiently interferes with chitin-triggered immune responses, such as defence gene induction and reactive oxygen species production in Medicago truncatula. Although RiSLM also binds to symbiotic (lipo)chitooligosaccharides it does not interfere significantly with symbiotic signalling in Medicago. Host-induced gene silencing of RiSLM greatly reduces fungal colonization levels. Taken together, our results reveal a key role for AM fungal LysM effectors to subvert chitin-triggered immunity in symbiosis, pointing to a common role for LysM effectors in both symbiotic and pathogenic fungi.


Quick Prostate Test (QPT): motion for a tool for the active contribution of the general pratictioner to the diagnosis and follow up of benign prostatic hyperplasia.
Less than 40% of men with LUTS consult their doctor. Patients consider the LUTS as physiological and are resigned to endure them. It is necessary to foster awareness of the micturition disorders, to monitor their development and to assess the effectiveness of therapies. At present the only validated test is the IPSS-Q8, but in Italy it is used by only 4% of General Practitioners (GPs). Because the IPSS is complex and not easy to handle, we need a more simple test but nevertheless efficient. The Italian Society of Urology (SIU) and the Italian Society for Interdisciplinary Primary Care (SIICP) presented the "Quick Prostate Test" (QPT) in November 2012. We aimed to evaluate the efficiency of QPT versus the IPSSQ8 and its suitability in primary care. The QPT is composed of 3 questions to be answered "yes" or "no." The answer "yes" just to one question makes "positive" the test. We enrolled 64 men, ≥ 50 years old, suffering from BPH, extracted from the database of five GPs. The patients were randomized into two arms: to the arm 1 only QPT was administered, to verify efficiency of the test; to the arm 2 both the QPT that the IPSS-Q8 were administered. Into the arm 1, the 96.4% has tested positive for QPT. Into the arm 2, the 89% of patients with one or two positive responses to the QPT showed a moderate IPSSQ8 score; the 75% of the patients with three positive responses to the QPT had a serious IPSS-Q8 score. The GPs (80%) have expressed the highest level of satisfaction for the QPT for the "time of administration" and for the "simplicity" of the test. The experience with the QPT has shown that the test is efficient and suitable in the primary care setting. We want to encourage the GPs to use the QPT for the monitoring of patients with lower urinary tract symptoms (LUTS) and to contribute to the validation of the test.


Hidden Historical Habitat-Linked Population Divergence and Contemporary Gene Flow of a Deep-Sea Patellogastropod Limpet.
Hydrothermal vents and hydrocarbon seeps in the deep ocean are rare oases fueled by chemosynthesis. Biological communities inhabiting these ecosystems are often distributed in widely separated habitats, raising intriguing questions on how these organisms achieve connectivity and whether habitat types facilitate intraspecific divergence. The deep-sea patellogastropod limpet Bathyacmaea nipponica that colonizes both vents and seeps across ∼2,400 km in the Northwest Pacific is a feasible model to answer these questions. We analyzed 123 individuals from four vents and three seeps using a comprehensive method incorporating population genomics and physical ocean modeling. Genome survey sequencing and genotyping-by-sequencing resulted in 9,838 single-nucleotide polymorphisms for population genomic analyses. Genetic divergence and demographic analyses revealed four habitat-linked (i.e., three seep and one vent) genetic groups, with the vent genetic group established via the opportunistic invasion of a few limpet larvae from a nearby seep genetic group. TreeMix analysis uncovered three historical seep-to-vent migration events. ADMIXTURE and divMigrate analyses elucidated weak contemporary gene flow from a seep genetic group to the vent genetic group. Physical ocean modeling underlined the potential roles of seafloor topography and ocean currents in shaping the genetic connectivity, contemporary migration, and local hybridization of these deep-sea limpets. Our study highlighted the power of integrating genomic and oceanographic approaches in deciphering the demography and diversification of deep-sea organisms. Given the increasing anthropogenic activities (e.g., mining and gas hydrate extraction) affecting the deep ocean, our results have implications for the conservation of deep-sea biodiversity and establishment of marine protected areas.


Rat liver mitochondrial monoamine oxidase. A change in the reaction mechanism on solubilization.
1. The kinetics of benzylamine oxidation by a soluble preparation of rat liver mitochondrial monoamine oxidase were investigated and were shown to conform to adouble-displacement (or Ping Pong) mechanism. 2. The pathway differs in detail from that followed by other amine oxidases, including the membrane-bound enzyme in rat liver mitochondrial outer membranes. 3. It is suggested taht the conformation of the protein in the soluble state differs from that in the membrane-bound state. 4. The full rate equations for this mechanism have been deposited as Supplementary Publication SUP 50039 (5pages) at the British Library (lending Division) (formely the National Lending Library for Science and Technology), Boston Spa, Yorks, LS237BQ, U.K.. from whom copies can be obtained on the terms indicated in Biochem. J (1975) 145,5.


Introducing a New Mobile Electroencephalography System and Evaluating Its Quality in Comparison to Clinical Electroencephalography.
Electroencephalography (EEG) is a crucial tool in cognitive neuroscience, enabling the study of neurophysiological function by measuring the brain's electrical activity. Its applications include perception, learning, memory, language, decision making and neural network mapping. Recently, interest has surged in extending EEG measurements to domestic environments. However, the high costs associated with traditional laboratory EEG systems have hindered accessibility for many individuals and researchers in education, research, and medicine. To tackle this, a mobile-EEG device named "DreamMachine" was developed. A more affordable alternative to both lab-based EEG systems and existing mobile-EEG devices. This system boasts 24 channels, 24-bit resolution, up to 6 h of battery life, portability, and a low price. Our open-source and open-hardware approach empowers cognitive neuroscience, especially in education, learning, and research, opening doors to more accessibility. This paper introduces the DreamMachine's design and compares it with the lab-based EEG system "asalabTM" in an eyes-open and eyes-closed experiment. The Alpha band exhibited higher power in the power spectrum during eyes-closed conditions, whereas the eyes-open condition showed increased power specifically within the Delta frequency range. Our analysis confirms that the DreamMachine accurately records brain activity, meeting the necessary standards when compared to the asalabTM system.


[Sonographic detection of fistulas in Crohn disease].
The sonographic detection of fistulas in Crohn's disease has been almost impossible up to now and had therefore not been attempted. Two findings of an enterocutaneous and a blind-ending colohepatic fistula are demonstrated. Probably sonographic diagnosis of fistulas is better than has been believed so far, due to the high contrast of tiny gas bubbles; the "compression technique" to bypass meteoristic intestinal loops, which is been increasingly employed, and the improved physical resolution that allows to distinguish between different layers of the intestinal wall, are further contributing factors to the increasing use of sonography in this category of diagnostics.


2,4-Substituted bispidines as rigid hosts for versatile applications: from κ-opioid receptor to metal coordination.
Bispidones (3,7-diazabicyclo[3.3.1]nonan-9-one) are bicyclic analogues of the natural antiarrhythmic agent, spartein. They can straightforwardly be obtained from two successive Mannich reactions. Reduction of the ketone gives the corresponding bispidol. Substituted bispidones and bispidols offer a large playground by varying the substituents, the configuration of the carbon atoms in position 2 and 4 as well as the conformation of the bicycle. While chair-boat conformers display a strong affinity for κ-opioid receptors, chair-chair bispidines provide adaptable coordination spheres for transition metal and rare-earth ions. Because of their very rich coordination chemistry, substituted bispidines have emerged in various applications of coordination chemistry, such as catalysis, magnetism and medical imaging.


Cellular vascular endothelial growth factor is a predictor of outcome in patients with acute myeloid leukemia.
Vascular endothelial growth factor (VEGF) is a potent mitogen for vascular endothelial cells. It has been associated with angiogenesis, growth, dissemination, metastasis, and poor outcome in solid tumors. To assess cellular VEGF levels and their prognostic significance in newly diagnosed acute myeloid leukemia (AML), we used a radioimmunoassay (RIA) to quantify VEGF levels in stored samples obtained before treatment from 99 patients with newly diagnosed AML treated at the MD Anderson Cancer Center from 1996 to 1998. Outcome in the 99 patients was representative of that observed in all patients seen at this institution with this diagnosis during these years, but the 99 patients had higher white blood cell (WBC) and blast counts than the other patients. Results of the RIA were confirmed by Western blot. There was a relationship between increasing VEGF levels and shorter survival (P =.01), as well as shorter disease-free survival, both from start of treatment and from complete response (CR) date. In contrast, there was no relationship between VEGF level and WBC or blast count, or between VEGF level and such established prognostic factors as age, cytogenetics, performance status, or presence of an antecedent hematologic disorder, and multivariate analysis indicated that VEGF was still prognostic for the above outcomes after accounting for these factors, as well as treatment. Our results suggest that at least in AML patients with higher WBC and blast counts, cellular VEGF level is an independent predictor of outcome.


A putative relay circuit providing low-threshold mechanoreceptive input to lamina I projection neurons via vertical cells in lamina II of the rat dorsal horn.
Lamina I projection neurons respond to painful stimuli, and some are also activated by touch or hair movement. Neuropathic pain resulting from peripheral nerve damage is often associated with tactile allodynia (touch-evoked pain), and this may result from increased responsiveness of lamina I projection neurons to non-noxious mechanical stimuli. It is thought that polysynaptic pathways involving excitatory interneurons can transmit tactile inputs to lamina I projection neurons, but that these are normally suppressed by inhibitory interneurons. Vertical cells in lamina II provide a potential route through which tactile stimuli can activate lamina I projection neurons, since their dendrites extend into the region where tactile afferents terminate, while their axons can innervate the projection cells. The aim of this study was to determine whether vertical cell dendrites were contacted by the central terminals of low-threshold mechanoreceptive primary afferents. We initially demonstrated contacts between dendritic spines of vertical cells that had been recorded in spinal cord slices and axonal boutons containing the vesicular glutamate transporter 1 (VGLUT1), which is expressed by myelinated low-threshold mechanoreceptive afferents. To confirm that the VGLUT1 boutons included primary afferents, we then examined vertical cells recorded in rats that had received injections of cholera toxin B subunit (CTb) into the sciatic nerve. We found that over half of the VGLUT1 boutons contacting the vertical cells were CTb-immunoreactive, indicating that they were of primary afferent origin. These results show that vertical cell dendritic spines are frequently contacted by the central terminals of myelinated low-threshold mechanoreceptive afferents. Since dendritic spines are associated with excitatory synapses, it is likely that most of these contacts were synaptic. Vertical cells in lamina II are therefore a potential route through which tactile afferents can activate lamina I projection neurons, and this pathway could play a role in tactile allodynia.


Functional characterization of group Ⅱ interferon, IFNf in the acipenseriform fish, Chinese sturgeon (Acipenser sinensis).
Teleost fish possess a diversity of type Ⅰ interferons (IFNs) repertoire, which play a crucial role in antiviral and antimicrobial immune responses. In our previous study, IFNe1-3 and IFNb were identified and cloned from Chinese sturgeon (Acipenser sinensis), an acipenseriform fish. However, the absence of Chinese sturgeon genome data has left the question of whether there are other type Ⅰ IFN members in this species unresolved. In this study, we have identified and characterized a novel IFN, IFNf in Chinese sturgeon (AsIFNf). Bioinformatics analysis revealed that the AsIFNf contains a unique disulfide bond (2 cysteines) located in the second exon and fifth exon region, distinguishing it from other reported teleost type I IFNs. Meanwhile, qPCR results showed that AsIFNf mRNA was detectable in all examined tissues and up-regulated in the spleen or kidney in response to poly I: C, Citrobacter freundii, and Spring Viremia of Carp Virus (SVCV), but not by LPS. Furthermore, compared to recombinant AsIFNe2 protein (rAsIFNe2), rAsIFNf exhibited a stronger protective effect on Chinese sturgeon fin cells against SVCV and also induced higher expression of antiviral genes Mx and viperin. Importantly, AsIFNf displayed characteristics similar to antimicrobial peptides (AMPs) with a positive charge and demonstrated a broad spectrum of antimicrobial activity in vitro. These findings provide a theoretical foundation for understanding the primitive structure and function of interferon, as well as deepening our comprehension of the innate immune system and disease defense in the endangered Chinese sturgeon.


Combined hereditary and somatic mutations of replication error repair genes result in rapid onset of ultra-hypermutated cancers.
DNA replication-associated mutations are repaired by two components: polymerase proofreading and mismatch repair. The mutation consequences of disruption to both repair components in humans are not well studied. We sequenced cancer genomes from children with inherited biallelic mismatch repair deficiency (bMMRD). High-grade bMMRD brain tumors exhibited massive numbers of substitution mutations (>250/Mb), which was greater than all childhood and most cancers (>7,000 analyzed). All ultra-hypermutated bMMRD cancers acquired early somatic driver mutations in DNA polymerase ɛ or δ. The ensuing mutation signatures and numbers are unique and diagnostic of childhood germ-line bMMRD (P < 10(-13)). Sequential tumor biopsy analysis revealed that bMMRD/polymerase-mutant cancers rapidly amass an excess of simultaneous mutations (∼600 mutations/cell division), reaching but not exceeding ∼20,000 exonic mutations in <6 months. This implies a threshold compatible with cancer-cell survival. We suggest a new mechanism of cancer progression in which mutations develop in a rapid burst after ablation of replication repair.


Notch signaling pathway is activated in motoneurons of spinal muscular atrophy.
Spinal muscular atrophy (SMA) is a neurodegenerative disease produced by low levels of Survival Motor Neuron (SMN) protein that affects alpha motoneurons in the spinal cord. Notch signaling is a cell-cell communication system well known as a master regulator of neural development, but also with important roles in the adult central nervous system. Aberrant Notch function is associated with several developmental neurological disorders; however, the potential implication of the Notch pathway in SMA pathogenesis has not been studied yet. We report here that SMN deficiency, induced in the astroglioma cell line U87MG after lentiviral transduction with a shSMN construct, was associated with an increase in the expression of the main components of Notch signaling pathway, namely its ligands, Jagged1 and Delta1, the Notch receptor and its active intracellular form (NICD). In the SMNΔ7 mouse model of SMA we also found increased astrocyte processes positive for Jagged1 and Delta1 in intimate contact with lumbar spinal cord motoneurons. In these motoneurons an increased Notch signaling was found, as denoted by increased NICD levels and reduced expression of the proneural gene neurogenin 3, whose transcription is negatively regulated by Notch. Together, these findings may be relevant to understand some pathologic attributes of SMA motoneurons.


The role of areas MT+/V5 and SPOC in spatial and temporal control of manual interception: an rTMS study.
Manual interception, such as catching or hitting an approaching ball, requires the hand to contact a moving object at the right location and at the right time. Many studies have examined the neural mechanisms underlying the spatial aspects of goal-directed reaching, but the neural basis of the spatial and temporal aspects of manual interception are largely unknown. Here, we used repetitive transcranial magnetic stimulation (rTMS) to investigate the role of the human middle temporal visual motion area (MT+/V5) and superior parieto-occipital cortex (SPOC) in the spatial and temporal control of manual interception. Participants were required to reach-to-intercept a downward moving visual target that followed an unpredictably curved trajectory, presented on a screen in the vertical plane. We found that rTMS to MT+/V5 influenced interceptive timing and positioning, whereas rTMS to SPOC only tended to increase the spatial variance in reach end points for selected target trajectories. These findings are consistent with theories arguing that distinct neural mechanisms contribute to spatial, temporal, and spatiotemporal control of manual interception.


Prioritizing and Implementing Primary Care Performance Measures for Ontario.
In the fall of 2014, Health Quality Ontario released A Primary Care Performance Measurement Framework for Ontario. Recognizing the large number of recommended measures and the limited availability of data related to those measures, the Steering Committee for the Primary Care Performance Measurement (PCPM) initiative established a prioritization process to select two subsets of high-value performance measures - one at the system level and one at the practice level. This article describes the prioritization process and its results and outlines the initiatives that have been undertaken to date to implement the PCPM framework and to advance primary care performance measurement and reporting in Ontario. Establishing a framework for primary care measurement and prioritizing system- and practice-level measures are essential steps toward system improvement. Our experience suggests that the process of implementing a performance measurement system is inevitably non-linear and incremental.


Treatment of premenstrual tension with lithium carbonate. A pilot study.
Some previous reports have suggested that lithium treatment is of benefit to women with the premenstrual tension syndrome (PMTS). In this study 15 women carefully selected for severe PMTS were given lithium carbonate (600-900 mg/day) continuously for three menstrual cycles. Lithium did not affect physical premenstrual symptoms and was ineffective in most women against behavioral premenstrual symptoms. Despite the low dosage most women also experienced significant drug-related side effects. Although a statistically significant improvement was recorded by several symptom rating instruments, this benefit was of no practical clinical value. The three women who responded best to lithium, and who requested continued treatment beyond 3 months, met diagnostic criteria for subsyndromal affective (cyclothymic) disorder. Lithium is not recommended for the average woman with PMTS.


Haploinsufficiency of the transcription factors FOXC1 and FOXC2 results in aberrant ocular development.
Anterior segment developmental disorders, including Axenfeld-Rieger anomaly (ARA), variably associate with harmfully elevated intraocular pressure (IOP), which causes glaucoma. Clinically observed dysgenesis does not correlate with IOP, however, and the etiology of glaucoma development is not understood. The forkhead transcription factor genes Foxc1 (formerly Mf1 ) and Foxc2 (formerly Mfh1 ) are expressed in the mesenchyme from which the ocular drainage structures derive. Mutations in the human homolog of Foxc1, FKHL7, cause dominant anterior segment defects and glaucoma in various families. We show that Foxc1 (+/-)mice have anterior segment abnormalities similar to those reported in human patients. These abnormalities include small or absent Schlemm's canal, aberrantly developed trabecular meshwork, iris hypoplasia, severely eccentric pupils and displaced Schwalbe's line. The penetrance of clinically obvious abnormalities varies with genetic background. In some affected eyes, collagen bundles were half normal diameter, or collagen and elastic tissue were very sparse. Thus, abnormalities in extracellular matrix synthesis or organization may contribute to development of the ocular phenotypes. Despite the abnormalities in ocular drainage structures in Foxc1 (+/-)mice, IOP was normal in almost all mice analyzed, on all genetic backgrounds and at all ages. Similar abnormalities were found in Foxc2 (+/-)mice, but no disease-associated mutations were identified in the human homolog FKHL14 in 32 ARA patients. Foxc1 (+/-)and Foxc2 (+/-)mice are useful models for studying anterior segment development and its anomalies, and may allow identification of genes that interact with Foxc1 and Foxc2 (or FKHL7 and FKHL14 ) to produce a phenotype with elevated IOP and glaucoma.


'Treating Africans differently': using skin colour as proxy for HIV risk.
This paper examines the qualitative results of two studies of Black African new settler communities in New Zealand. The analysis investigates the issues of stigma and microaggressions and their effects on Black African communities. Previous work that investigated experiences of stigma faced by relatively early Black African new settlers to New Zealand found that new settlers experienced stigma, and their resulting isolation had negative implications for access to health and social services, regardless of HIV status. This paper is a meta-analysis of two published studies, using original qualitative data from each. Researchers for the first study, Standing in the Fire, interviewed 13 Black Africans living with HIV who were new settlers to New Zealand. The second study, AfricaNZ Care, was a national mixed method study which surveyed 703 Black African new settlers, and included 131 participants in 23 different focus groups. The present analysis includes only the qualitative data from the second study. Some, but not most, participants of the second study were living with HIV. Black African identity is used as a proxy for HIV status among non-HIV specialist health care workers. Participants reported experiences of stigma and microaggressions based on their race, and a lack of knowledge about HIV in non-HIV specialist nurses and other health care workers. They also experienced poor health care and education practices, professional prejudice against colleagues living with HIV and institutional challenges including failure to protect patient confidentiality. HIV-related stigma and microaggressions remain significant problems, not only for people living with HIV but also for those perceived to be at high risk for HIV. Experiences of stigmatising behaviour by health care professionals have a broad and significant range of impacts on patients' health outcomes.


The effect of mandala coloring on pregnancy-related anxiety.
In the literature, the efficacy of mandala coloring on pregnancy-related anxiety has been investigated, and no study has been found on its effect on anxiety. This randomized controlled trial was designed to assess the efficacy of mandala coloring application on pregnancy-related anxiety. The study included 77 primiparous pregnant women: 39 in the control group and 38 in the experimental group, and was carried out in an education-research hospital's obstetric clinic. Data for the study were gathered using Information Form, the Pregnancy-related Anxiety Scale, and the State Anxiety Inventory. While the experimental group did mandala coloring during a month twice in a week, and the control group was not do anything. The "Pregnancy-related Anxiety Scale" and "The State Anxiety Inventor" were used to assess anxiety in all groups before and after application. In the statistical analysis of the data, the SPSS 23.0 package program was used. In all tests, p < 0.005 was considered statistically significant. Immediately post-intervention, the women in the experimental group had a significantly higher reduce pregnancy-related anxiety (p < 0.05). Especially in the mandala group within according to control group were found to have significantly reduced anxiety the application (PrAS-1 82,50 ± 2,47 vs. 84,41 ± 4,12; PrAS-2 77,92 ± 2,16 vs. 83,89 ± 3,80; PrAS-3 72,05 ± 3,97 vs. 84,10 ± 4,35; PrAS-Total 77,49 ± 5,21 vs. 84,13 ± 4,07; STAI-1 52,39 ± 2,50 vs. 51,17 ± 1,16; STAI-2 49,73 ± 0,75 vs. 49,05 ± 1,27; STAI-3 48,84 ± 3,17 vs. 49,02 ± 1,22; STAI-Total 50,32 ± 2,8 vs. 49,75 ± 1,58). Mandala coloring is effective in reducing pregnancy-related anxiety. (NCT05534867).


Toward a blueprint for regeneration.
Tissue regeneration has been studied for hundreds of years, yet remains one of the less understood topics in developmental biology. The recent Keystone Symposium on Mechanisms of Whole Organ Regeneration brought together biologists, clinicians and bioengineers representing an impressive breadth of model systems and perspectives. Members of the growing regeneration community discussed classic and new ideas on mechanisms of regeneration and how these can be applied to regenerative medicine.


Age-related inflammatory bone marrow microenvironment induces ineffective erythropoiesis mimicking del(5q) MDS.
Anemia is characteristic of myelodysplastic syndromes (MDS). The mechanisms of anemia in MDS are unclear. Using a mouse genetic approach, here we show that dual deficiency of mDia1 and miR-146a, encoded on chromosome 5q and commonly deleted in MDS (del(5q) MDS), causes an age-related anemia and ineffective erythropoiesis mimicking human MDS. We demonstrate that the ageing bone marrow microenvironment is important for the development of ineffective erythropoiesis in these mice. Damage-associated molecular pattern molecules (DAMPs), whose levels increase in ageing bone marrow, induced TNFα and IL-6 upregulation in myeloid-derived suppressor cells (MDSCs) in mDia1/miR-146a double knockout mice. Mechanistically, we reveal that pathologic levels of TNFα and IL-6 inhibit erythroid colony formation and differentially affect terminal erythropoiesis through reactive oxygen species-induced caspase-3 activation and apoptosis. Treatment of the mDia1/miR-146a double knockout mice with all-trans retinoic acid, which promoted the differentiation of MDSCs and ameliorated the inflammatory bone marrow microenvironment, significantly rescued anemia and ineffective erythropoiesis. Our study underscores the dual roles of the ageing microenvironment and genetic abnormalities in the pathogenesis of ineffective erythropoiesis in del(5q) MDS.


Nanoscale symmetry fluctuations in ferroelectric barium titanate, BaTiO3.
Crystal charge density is a ground-state electronic property. In ferroelectrics, charge is strongly influenced by lattice and vice versa, leading to a range of interesting temperature-dependent physical properties. However, experimental determination of charge in ferroelectrics is challenging because of the formation of ferroelectric domains. Demonstrated here is the scanning convergent-beam electron diffraction (SCBED) technique that can be simultaneously used for imaging ferroelectric domains and identifying crystal symmetry and its fluctuations. Results from SCBED confirm the acentric tetragonal, orthorhombic and rhombohedral symmetry for the ferroelectric phases of BaTiO3. However, the symmetry is not homogeneous; regions of a few tens of nanometres retaining almost perfect symmetry are interspersed in regions of lower symmetry. While the observed highest symmetry is consistent with the displacive model of ferroelectric phase transitions in BaTiO3, the observed nanoscale symmetry fluctuations are consistent with the predictions of the order-disorder phase-transition mechanism.


Gastrointestinal surgery for severe obesity.
The National Institutes of Health Consensus Development Conference on Gastrointestinal Surgery for Severe Obesity brought together surgeons, gastroenterologists, endocrinologists, psychiatrists, nutritionists, and other health care professionals as well as the public to address: the nonsurgical treatment options for severe obesity, the surgical treatments for severe obesity and the criteria for selection, the efficacy and risks of surgical treatments for severe obesity, and the need for future research on and epidemiological evaluation of these therapies. Following 2 days of presentations by experts and discussion by the audience, a consensus panel weighed the evidence and prepared their consensus statement. Among their findings, the panel recommended that (1) patients seeking therapy for severe obesity for the first time should be considered for treatment in a nonsurgical program with integrated components of a dietary regimen, appropriate exercise, and behavioral modification and support, (2) gastric restrictive or bypass procedures could be considered for well-informed and motivated patients with acceptable operative risks, (3) patients who are candidates for surgical procedures should be selected carefully after evaluation by a multidisciplinary team with medical, surgical, psychiatric, and nutritional expertise, (4) the operation be performed by a surgeon substantially experienced with the appropriate procedures and working in a clinical setting with adequate support for all aspects of management and assessment, and (5) lifelong medical surveillance after surgical therapy is a necessity. The full text of the consensus panel's statement follows.


Follicle growth, corpus luteum function and their effects on embryo development in postpartum dairy cows.
Absent or irregular ovarian cycles in lactating dairy cows are caused by failure to ovulate the dominant follicle at the appropriate time. The follicle then either regresses or develops into a cyst. This process can be triggered by a variety of metabolic and disease factors that act at the hypothalamus and pituitary gland to inhibit pulsatile LH secretion and the LH surge, and at the ovary to reduce follicular growth and oestradiol production. Cows of poor energy status have low circulating concentrations of insulin-like growth factor I (IGF-I). Predisposing factors include calving difficulties, inappropriate diet, reduced intake of dry matter and a high rate of body condition score loss. Various stressors predispose the follicle to cyst development by inhibiting the LH surge and ovulation; these include common infections, such as mastitis. Even when ovulation does occur, poor follicular development may result in production of an inadequate corpus luteum. The timing of the increase in progesterone in the early luteal phase (days 4-5) appears to be a key determinant of fertility, probably because it alters the secretory activity of the reproductive tract, thus influencing embryonic growth and interferon-tau production. A period of negative energy balance after calving can reduce fertility even though metabolic parameters have apparently improved at the time of service.


Primary umbilical adenocarcinoma. A case report and review of literature.
A case of primary adenocarcinoma of the umbilicus is reported along with a review of the literature and discussion of possible origins of glandular tissue within this area. Multiple connections are present within the umbilicus (vascular, lymphatic, and embryologic) that may give this area access to metastatic lesions. The primary tumors may originate within the usual umbilical tissue (skin and soft tissue). Glands that are not normally present in the region of the umbilicus rarely develop malignant neoplasms. As postulated, glandular tissue may arise either as metaplasia from squamous epithelium or from glandular embryologic rests including omphalomesenteric duct remnants and urachal remnants.


Trees' role in nitrogen leaching after organic, mineral fertilization: a greenhouse experiment.
New sustainable agriculture techniques are arising in response to the environmental problems caused by intensive agriculture, such as nitrate leaching and surface water eutrophication. Organic fertilization (e.g., with sewage sludge) and agroforestry could be used to reduce nutrient leaching. We assessed the efficiency of establishing trees and pasture species in environmentally sensitive, irrigated Mediterranean grassland soils in controlling nitrate leaching. Four vegetation systems-bare soil, pasture species, cherry trees [ (L.) L.], and pasture-tree mixed plantings-and five fertilization treatments-control, two doses of mineral fertilizer, and two doses of organic fertilizer (sewage sludge)-were tested in a greenhouse experiment over 2 yr. In the experiment, the wet and warm climate characteristics of Mediterranean irrigated croplands and the plant-to-plant and soil-to-plant interactions that occur in open-field agroforestry plantations were simulated. Following a factorial design with six replicates, 120 pots (30-cm radius and 120 cm deep) were filled with a sandy, alluvial soil common in the cultivated fluvial plains of the region. The greatest pasture production and tree growth were obtained with sewage sludge application. Both pasture production and tree growth decreased significantly in the pasture-tree mixed planting. Nitrate leaching was negligible in this latter treatment, except under the highest dose of sewage sludge application. The rapid mineralization of sludge suggested that this organic fertilizer should be used very cautiously in warm, irrigated Mediterranean soils. Mixed planting of pasture species and trees, such as , could be a useful tool for mitigating nitrate leaching from irrigated Mediterranean pastures on sandy soils.


dpy-18 encodes an alpha-subunit of prolyl-4-hydroxylase in caenorhabditis elegans.
Collagen is an extracellular matrix (ECM) component encoded by a large multigene family in multicellular animals. Procollagen is post-translationally modified by prolyl-4-hydroxylase (EC 220.127.116.11) before secretion and participation in ECM formation. Therefore, collagen processing and regulation can be studied by examining this required interaction of prolyl-4-hydroxylase with procollagen. High-resolution polymorphism mapping was used to place the Caenorhabditis elegans dpy-18 gene on the physical map, and we show that it encodes a prolyl-4-hydroxylase alpha catalytic subunit. The Dpy phenotype of dpy-18(e364) amber mutants is more severe when this mutation is in trans to the noncomplementing deficiency tDf7, while the dpy-18(e499) deletion mutant exhibits the same phenotype as dpy-18(e499)/tDf7. Furthermore, dpy-18 RNA interference (RNAi) in wild-type worms results in Dpy progeny, while dpy-18 (RNAi) in dpy-18(e499) mutants does not alter the Dpy phenotype of their progeny. These observations suggest that the dpy-18 null phenotype is Dpy. A dpy-18::gfp promoter fusion construct is expressed throughout the hypodermis within the cells that abundantly produce the cuticle collagens, as well as in certain head and posterior neurons. While prolyl-4-hydroxylase has been studied extensively by biochemical techniques, this is the first report of a mutationally defined prolyl-4-hydroxylase in any animal.


Optimal nutrition: calcium, magnesium and phosphorus.
In the past, a major challenge for nutrition research was in defining indicators of nutritional adequacy. More recently, the research base related to the role of nutrition in chronic disease has expanded sufficiently to permit moving beyond deficiency indicators to other indicators with broader functional significance. Thus, nutrition research is faced with the new challenge of defining 'optimal nutrition'. One definition of optimal nutrition with respect to any particular nutrient could be when a functional marker reaches an 'optimal value' or plateau beyond which it is not longer affected by intake or stores of the nutrient. A functional marker of nutrient status could be defined as a physiological or biochemical factor which (1) is related to function or effect of the nutrient in target tissue(s) and (2) is affected by dietary intake or stores of the nutrient (which may include markers of disease risk). Examples of such indicators or markers are those related to risk of chronic diseases such as osteoporosis, CHD, or hypertension. The present review focuses on the concept of optimal nutrition with respect to three nutrients, Ca, Mg and P. However, for P and Mg there are as yet no functional indicators which respond to dietary intake, and in such cases nutrient requirements are established using more traditional approaches, e.g. balance data. For Ca, there has been interest in using maximal Ca retention, which is based on balance data, bone mass measurements and biomarkers of bone turnover as useful functional indicators of the adequacy of Ca intake.


Bladder incarceration with perforation in scrotal herniation: A case report.
Inguinoscrotal hernias containing urinary bladder are very rare. There are only a few cases described with perforation in the scrotum. This illness is a severe and should be kept in mind with any patient complaining of a scrotal hernia. We report a case of bladder perforation caused by inguinoscrotal hernia with incarceration and severe sepsis in a 93-year-old male. A computed tomography scan with excretory phase was used to diagnose the disease. A laparotomy with partial bladder resection and herniotomy were performed. Our patient recovered well from surgery. Ten days after surgery, a cystography was performed showing no extravasation.


COVID-19 in people with neurofibromatosis 1, neurofibromatosis 2, or schwannomatosis.
People with pre-existing conditions may be more susceptible to severe COVID-19 when infected by SARS-CoV-2. The relative risk and severity of SARS-CoV-2 infection in people with rare diseases such as neurofibromatosis type 1 (NF1), neurofibromatosis type 2 (NF2), or schwannomatosis (SWN) is unknown. We investigated the proportions of people with NF1, NF2, or SWN in the National COVID Cohort Collaborative (N3C) electronic health record data set who had a positive test result for SARS-CoV-2 or COVID-19. The cohort sizes in N3C were 2501 (NF1), 665 (NF2), and 762 (SWN). We compared these with N3C cohorts of patients with other rare diseases (98-9844 individuals) and the general non-NF population of 5.6 million. The site- and age-adjusted proportion of people with NF1, NF2, or SWN who had a positive test result for SARS-CoV-2 or COVID-19 (collectively termed positive cases) was not significantly higher than in individuals without NF or other selected rare diseases. There were no severe outcomes reported in the NF2 or SWN cohorts. The proportion of patients experiencing severe outcomes was no greater for people with NF1 than in cohorts with other rare diseases or the general population. Having NF1, NF2, or SWN does not appear to increase the risk of being SARS-CoV-2 positive or of being a patient with COVID-19 or of developing severe complications from SARS-CoV-2.


Human Gut Microbiota in Coronary Artery Disease: A Systematic Review and Meta-Analysis.
In recent years, the importance of the gut microbiome in human health and disease has increased. Growing evidence suggests that gut dysbiosis might be a crucial risk factor for coronary artery disease (CAD). Therefore, we conducted a systematic review and meta-analysis to determine whether or not CAD is associated with specific changes in the gut microbiome. The V3-V4 regions of the 16S rDNA from fecal samples were analyzed to compare the gut microbiome composition between CAD patients and controls. Our search yielded 1181 articles, of which 21 met inclusion criteria for systematic review and 7 for meta-analysis. The alpha-diversity, including observed OTUs, Shannon and Simpson indices, was significantly decreased in CAD, indicating the reduced richness of the gut microbiome. The most consistent results in a systematic review and meta-analysis pointed out the reduced abundance of Bacteroidetes and Lachnospiraceae in CAD patients. Moreover, Enterobacteriaceae, Lactobacillus, and Streptococcus taxa demonstrated an increased trend in CAD patients. The alterations in the gut microbiota composition are associated with qualitative and quantitative changes in bacterial metabolites, many of which have pro-atherogenic effects on endothelial cells, increasing the risk of developing and progressing CAD.


Audism: exploring the metaphysics of oppression.
This article traces the development of the concept of "audism" from its inception in the mid-1970s by exploring three distinct dimensions of oppression: individual, institutional, and metaphysical. Although the first two aspects of audism have been identified, there is a deeply rooted belief system regarding language and human identity that is yet to be explored within the context of audism. This article attempts to expose how our particular historical and philosophical constructions of language and being have created what French philosopher Jacques Derrida calls phonocentrism. Although Derrida does not discuss audism, his deconstruction of the Western notion of language provides a lens through which we can better see the orientation that has provided fertile ground out of which individual and institutional audism has flourished.


Peripheral chemoreceptors in health and disease.
Peripheral chemoreceptors (carotid and aortic bodies) detect changes in arterial blood oxygen and initiate reflexes that are important for maintaining homeostasis during hypoxemia. This mini-review summarizes the importance of peripheral chemoreceptor reflexes in various physiological and pathophysiological conditions. Carotid bodies are important for eliciting hypoxic ventilatory stimulation in humans and in experimental animals. In the absence of carotid bodies, compensatory upregulation of aortic bodies as well as other chemoreceptors contributes to the hypoxic ventilatory response. Peripheral chemoreceptors are critical for ventilatory acclimatization at high altitude. They also contribute in part to the exercise-induced hyperventilation, especially with submaximal and heavy exercise. During pregnancy, hypoxic ventilatory sensitivity increases, perhaps due to the actions of estrogen and progesterone on chemoreceptors. Augmented peripheral chemoreceptors have been implicated in early stages of recurrent apneas, congestive heart failure, and certain forms of hypertension. It is likely that chemoreceptors tend to maintain oxygen homeostasis and act as a defense mechanism to prevent the progression of the morbidity associated with these diseases. Experimental models of recurrent apneas, congestive heart failure, and hypertension offer excellent opportunities to unravel the cellular mechanisms associated with altered chemoreceptor function.


[Iodide enrichment in malignant tumours after gene transfer].
After the cloning of the gene encoding the sodium-iodide symporter several trials were made to develop a radioiodine treatment for multiple tumour entities based on NIS gene transfer. These studies revealed in vitro as well as in vivo a tremendous enhancement of iodide accumulation, which was followed by a rapid efflux. Therapy effects were observed in vitro by clonogenic assays and in vivo by growth inhibition of the treated tumours. However, the interpretation of these results were largely different. Problems of radioiodine therapy after NIS transfer are low efficiency of gene transfer and the short exposure time for the tumours caused by the rapid efflux. Trials to enhance therapeutic efficiency by co-transfer of the gene encoding thyroperoxidase failed due to the low enzyme activity.


Vaccination of swine with an inactivated porcine parvovirus vaccine in the presence of passive immunity.
A study was conducted to determine whether low hemagglutination inhibiting (HI) titers (1:5) for porcine parvovirus (PPV) block the development of immune response to a PPV vaccine. Pigs with low (1:5), medium (1:10 or 1:20), or high (1:40 or 1:80) titers were obtained by IV injections with various amounts of PPV immune serum. Pigs were inoculated with 1 or 2 doses of vaccine and were monitored for serum HI antibodies to PPV. Pigs with low titers responded to vaccine just as well as did the seronegative pigs. The HI titers of pigs with medium titers did not increase after first vaccination. After the second vaccination, however, their titers increased and were similar to those of pigs with low titers. High titers blocked the response to vaccination. The pigs that received 2 doses of vaccine had higher titers than did those of pigs that received 1 dose of vaccine. The results indicated that low titers, which would be expected in gilts at the time of vaccination, do not interfere with immunization by the inactivated PPV vaccine, and that 2 doses of vaccine may provide better and longer lasting immune response to inactivated PPV vaccine and probably longer lasting immunity against PPV-induced reproductive failure.


Retrospective PCR-based species identification of Leishmania in two patients with visceral leishmaniasis in Serbia.
Retrospective molecular identification of Leishmania parasites in two patients with visceral leishmaniasis (VL) previously treated in Serbia was carried out. DNA was isolated from unstained bone marrow smears (BMSs) kept for 11 and 8 years. Genus-specific real-time PCR was combined with conventional PCR and sequencing for detection and species identification. In 2003, a 40-year-old Serbian male was admitted to the Clinical Centre of Serbia (CCS) with fever, sweating, fatigue and splenomegaly, which developed over a period of 7 weeks. He had frequently travelled around Europe. VL was confirmed by microscopy of Giemsa-stained BMS. Treatment by pentavalent antimonials was successfully completed. Two years later, the patient developed post-kala-azar dermal leishmaniasis. Treatment resulted in symptom resolution. Later on, Leishmania infantum was identified as the causative agent of the VL by sequencing of the ITS (internal transcribed spacer) region; mixed Leishmania spp. infection could not be excluded. In 2006, a 33-year-old female from Vojvodina, Serbia, with pre-existing diabetes mellitus and chronic meningoencephalitis and a history of frequent visits to the Montenegrin seacoast, was admitted to the CCS with fever, pancytopenia and moderate hepatosplenomegaly. A stained BMS revealed abundant Leishmania amastigotes. Indirect haemagglutination analysis was positive with a titre of 1 : 2048, and a rapid dipstick rK39 test was also positive. Treatment by liposomal amphotericin B was successful; however, shortly after, the patient developed neural infection and pneumonia and died. The causative agent was identified as L. infantum. Molecular diagnosis of VL and species delineation using DNA from unstained BMSs stored for several years is possible.


Lyophilized hyperimmune equine serum as a source of antibodies for neonatal foals.
In a study with 15 neonatal foals (5 per treatment group), foals were fed within 4 hours of birth as follows: 250 ml of colostrum, 250 ml of lyophilized serum reconstituted at 5 times the original concentration, or 250 ml of a mixture (1:1) of colostrum and lyophilized serum. Foal serum samples were tested for immunoglobulin (Ig)G concentration and titrated for anti-equine rhinovirus 1 and anti-equine influenza A1 and A2 antibodies at 0 and 24 hours after foals were born. Except in a foal which had suckled the dam before treatment, there was no evidence of IgG or specific viral antibodies in the samples taken at birth. There were no significant differences found in the serum IgG concentrations and antibody titers among the 3 treatment groups. Seemingly, IgG was absorbed efficiently from both serum and colostrum, so that the use of reconstituted lyophilized serum as a prophylactic measure of conferring passive immunity to a newborn foal deserves serious consideration.


Agaritine from Agaricus blazei Murrill induces apoptosis in the leukemic cell line U937.
Agaricus blazei Murrill (ABM) has been shown to exhibit immunostimulatory and anti-cancer activities; however, its mechanism of action is poorly understood. We recently found that the diffusible fraction of hot-water extract of ABM exhibits anti-tumor activity toward leukemic cells, and identified it as agaritine, a hydrazine-containing compound. In the present study, we examined the morphological and cytochemical effects of agaritine on U937 cells to elucidate the tumoricidal mechanism of agaritine. Surface expression of phosphatidylserine (evaluated by annexin V binding), Fas antigen, DNA cleavage using TUNEL staining, changes in caspase activities and cytochrome c release, before and after treatment with agaritine, were examined using U937 cells. Nuclear damage, DNA fragmentation, was observed by Wright-Giemsa, TUNEL staining and agarose gel electrophoresis when U937 cells were incubated with 10μg/mL of agaritine for 48h. Flow cytometric analysis indicated that agaritine augments the proportion of annexin V-positive U937 cells without significant change in Fas antigen expression. Activities of caspase-3, -8 and -9 were gradually increased after the addition of agaritine. In the presence of caspase-3 or granzyme B inhibitor, except for the caspase-8 inhibitor, annexin V expression was significantly decreased, suggesting that mainly caspase-3 and -9 participate in the apoptotic pathway. Furthermore, cytochrome c release was detected by western blotting analysis after agaritine treatment. These results strongly suggest that the ABM constituent agaritine moderately induces apoptosis in U937 leukemic cells via caspase activation through cytochrome c release from mitochondria. This is the first report suggesting that the anti-tumor effect of agaritine is mediated through apoptosis. The present results might provide helpful suggestions for the design of anti-tumor drugs toward leukemia patients.


Phase 2 study of concurrent radiotherapy and temozolomide followed by temozolomide and lomustine in the treatment of children with high-grade glioma: a report of the Children's Oncology Group ACNS0423 study.
The prognosis for children with malignant glioma is poor. This study was designed to determine whether lomustine and temozolomide following radiotherapy and concurrent temozolomide improves event-free survival (EFS) compared with historical controls with anaplastic astrocytoma (AA) or glioblastoma (GBM) and whether survival is influenced by the expression of O6-methylguanine-DNA-methyltransferase (MGMT). Following maximal surgical resection, newly diagnosed children with nonmetastatic high-grade glioma underwent involved field radiotherapy with concurrent temozolomide. Adjuvant chemotherapy consisted of up to 6 cycles of lomustine 90 mg/m(2) on day 1 and temozolomide 160 mg/m(2)/day ×5 every 6 weeks. Among the 108 eligible patients with AA or GBM, 1-year EFS was 0.49 (95% CI, 0.39-0.58), similar to the original CCG-945-based design model. However, EFS and OS were significantly improved in ACNS0423 compared with the 86 AA or GBM participants treated with adjuvant temozolomide alone in the recent ACNS0126 study (1-sided log-rank P = .019 and .019, respectively). For example, 3-year EFS was 0.22 (95% CI, 0.14-0.30) in ACNS0423 compared with 0.11 (95% CI, 0.05-0.18) in ACNS0126. Stratifying the comparison by resection extent, the addition of lomustine resulted in significantly better EFS and OS in participants without gross-total resection (P = .019 and .00085 respectively). The difference in EFS and OS was most pronounced for participants with GBM (P = .059 and 0.051, respectively), and those with MGMT overexpression (P = .00036 and .00038, respectively). The addition of lomustine to temozolomide as adjuvant therapy in ACNS0423 was associated with significantly improved outcome compared with the preceding COG ACNS0126 HGG study in which participants received temozolomide alone.


[Predictors of osteoarticular problems in a cohort of computer users in Lebanon: about 810 employees].
The aim of the study is the analysis of osteoarticular problems (OAP) occurring in a group of computer users (CU) in search of correlation between symptoms and different etiological factors. Questionnaire of 31 items dealing with personal data, the activity of the CU, stress, the occurrence of osteoarticular problems during the last month (Oaplm) and last twelve months (Oaptm) and a checklist of 17 items covering the physical characteristics of the workplace. DESCRIPTIVE ANALYSIS * 810 respondents of mean age 36 +/- 9 years and predominantly female (69%) * seniority at the workplace (12.5 +/- 9 years), pace of work (825 +/- 1.5 hours/day and 5.5 +/- 1 days/ week), number of breaks (13 +/- 1.04/d), duration of breaks (35 min +/- 25/d), 44.5% in sports activities, work stress in 92% of participants * OAP described:--Osteoarticular problems last month (62%), neck pain (68%), shoulder (46%) and lumbar spine (62%) pain. Tingling hands (40%). Headache (55.5%). Temporomandibular disorders (18.5%)--Osteoarticular problems the last twelve months (46%). UNIVARIATE ANALYSIS: Detection of risk factors * RISK FACTORS and Oaplm relationship: female, weight gain, secretary, stress, pain during labor and work stoppages withp < 0.05 * RISK FACTORS and Oaptm relationship: the position of secretary, stress, pain at work, work stop-pages for Oaplm withp < 0.05 * Protection factor: sports more than one time per week. MULTIVARIATE ANALYSIS: Oaplm occurrent factors: weight gain, Oaptm withp < 0.05. Protection factor: well designed workstation * Oaptm occurrent factors: age, stress and Oaplm with p < 0.05. Significant prevalence of osteoarticular problems in Lebanese computer users. age, Oaptm, weight gain, stress, work-break cycle not respected and poor layout of the workstation. Ergonomic interventions are necessary and indispensable to reduce the cost of occupational diseases related to the CU, and ensure good mental and physical health.


Pilocarpine-induced seizures cause selective time-dependent changes to adult-generated hippocampal dentate granule cells.
Aberrantly interconnected granule cells are characteristic of temporal lobe epilepsy. By reducing network stability, these abnormal neurons may contribute directly to disease development. Only subsets of granule cells, however, exhibit abnormalities. Why this is the case is not known. Ongoing neurogenesis in the adult hippocampus may provide an explanation. Newly generated granule cells may be uniquely vulnerable to environmental disruptions relative to their mature neighbors. Here, we determine whether there is a critical period after neuronal birth during which neuronal integration can be disrupted by an epileptogenic insult. By bromodeoxyuridine birthdating cells in green fluorescent protein-expressing transgenic mice, we were able to noninvasively label granule cells born 8 weeks before (mature), 1 week before (immature), or 3 weeks after (newborn) pilocarpine-epileptogenesis. Neuronal morphology was examined 4 and 8 weeks after pilocarpine treatment. Strikingly, almost 50% of immature granule cells exposed to pilocarpine-epileptogenesis exhibited aberrant hilar basal dendrites. In contrast, only 9% of mature granule cells exposed to the identical insult possessed basal dendrites. Moreover, newborn cells were even more severely impacted than immature cells, with 40% exhibiting basal dendrites and an additional 20% exhibiting migration defects. In comparison, <5% of neurons from normal animals exhibited either abnormality, regardless of age. Together, these data demonstrate the existence of a critical period after the birth of adult-generated neurons during which they are vulnerable to being recruited into epileptogenic neuronal circuits. Pathological brain states therefore may pose a significant hurdle for the appropriate integration of newly born endogenous, and exogenous, neurons.


Analysis of the glycoprotein and protein composition of Bernard-Soulier platelets by single and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Previous reports have described conflicting results concerning the glycoprotein (GP) and protein composition of Bernard-Soulier platelets. In view of this controversy we have analyzed the platelets of four Bernard-Soulier patients using improved single and two-dimensional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis procedures. An absence of staining for carbohydrate of membrane GP Ib was characteristic for the platelets of each patient. Major periodate-Schiff staining bands corresponding to membrane GP IIb, IIIa, and IIIb were clearly detected and their presence was confirmed by two-dimensional SDS-polyacrylamide gel electrophoresis. The protein content of the Bernard-Soulier platelets was increased two- to fourfold. However, analysis of their protein composition using 7-12% acrylamide gradient gels showed normal polypeptide profiles. Lactoperoxidase-catalyzed 125I-labeling of the Bernard-Soulier platelet surface proteins was followed by SDS-polyacrylamide gel electrophoresis and autoradiography. No labeling in the Ib position was detected whereas the other major membrane GP, including Ia and IIa, were normally located. In contrast, GP Ib was clearly detected by periodate-Schiff staining and autoradiography when normal human platelets that had been exhaustively treated with neuraminidase before the lactoperoxidase-catalyzed iodination were analysed. No abnormalities were detected in the GP patterns of membranes isolated from the patients' erythrocytes. Only a severe molecular abnormality or possible deletion of GP Ib could account for this major platelet lesion in the Bernard-Soulier syndrome.


Specific membranous structures associated with the replication of group A arboviruses.
INTRACYTOPLASMIC MEMBRANOUS STRUCTURES OF A UNIQUE TYPE WERE ASSOCIATED WITH THE REPLICATION OF THREE GROUP A ARBOVIRUSES: Semliki Forest virus (SFV), Sindbis virus, or Western equine encephalomyelitis virus. The structures, referred to as type 1 cytopathic vacuoles (CPV-1), were membrane-limited and characteristically lined by regular membranous spherules measuring 50 nm in diameter. The membranous spherules typically contained a fine central density, but were neither virus cores nor virions. Detection of CPV-1 by electron microscopy at 3 to 6 hr postinfection coincided with the time of rapid virus growth and preceded the accumulation of virus nucleocapsids. A range of 20 to 100 CPV-1 profiles were counted per 100 ultrathin cell sections at 6 to 9 hr postinfection when viruses were grown in chick embryo, baby hamster kidney, or mouse L cells. Maximum counts remained in the same range even when the multiplicity of infection was varied over 100-fold. Inhibition of cellular ribonucleic acid (RNA) and protein synthesis by actinomycin D during SFV infection did not decrease the counts of CPV-1; however, biogenesis of CPV-1 was decreased when viral replication was limited by inhibitors of viral RNA synthesis (guanidine) or of viral protein synthesis (cycloheximide). On the basis of present and earlier findings, we concluded that formation of CPV-1 must result from a virus-specified modification of pre-existing host cell macromolecules.


Influence of microstructural variations on morphology and separation properties of polybutadiene-based polyurethanes.
Polybutadiene-based polyurethanes with different cis/trans/1,2-vinyl microstructure contents are synthesized. The phase morphology and physical properties of the polymers are investigated using spectroscopic analysis (FTIR and Raman), differential scanning calorimetry (DSC), X-ray scattering (WAXD and SAXS) and atomic force microscopy (AFM). In addition, their gas transport properties are determined for different gases at 4 bar and 25 °C. Thermodynamic incompatibility and steric hindrance of pendant groups are the dominant factors affecting the morphology and properties of the PUs. FTIR spectra, DSC, and SAXS analysis reveal a higher extent of phase mixing in high vinyl-content PUs. Moreover, the SAXS analysis and AFM phase images indicate smaller microdomains by increasing the vinyl content. Smaller permeable soft domains as well as the lower phase separation of the PUs with higher vinyl content create more tortuous pathways for gas molecules and deteriorate the gas permeability of the membranes.


Epidemiology and psychological factors of whiplash associated disorders in Japanese population.
[Purpose] This study was designed to examine the epidemiological background of Whiplash-associated disorders in Japanese adults and to investigate the psychological factors associated with prolonged treatment for Whiplash-associated disorders. [Subjects and Methods] An online survey was completed by 127,956 participants, of whom 4,164 had been involved in a traffic collision. A random sample of the collision participants (n=1,698) were provided with a secondary questionnaire. From the 974 (57.4%) participants who returned the questionnaire, 183 cases (intractable neck pain treated over a period of 6 months) and 333 controls (minor neck pain treated within 3 months) were selected. Among the control group, the psychological factors associated with prolonged treatment for Whiplash-associated disorders were investigated. [Results] Among the 4,164 collision participants, 1,571 (37.7%) had experienced Whiplash-associated disorders. The prevalence in the general population was 1.2% (1.3% in male and 1.0% in female). Significant differences were observed between the cases and controls for all psychological factors, although both groups had similar distributions of age and gender. [Conclusion] Poor psychological factors were associated with prolonged treatment for whiplash-associated disorders in Japanese adults. These psychological factors should be considered during the treatment of whiplash-associated disorders.


The racialized construction of exceptionality: Experimental evidence of race/ethnicity effects on teachers' interventions.
Scholars, policy-makers, and practitioners have long argued that students of color are over-represented in special education and under-represented in gifted education, arguing that educators make racially/ethnically biased decisions to refer and qualify students with disabilities and giftedness. Recent research has called this into question, focusing on the role of confounders of race/ethnicity. However, the role of educator decisions in the disproportionality is still unclear. In this study, I examine the role of student race/ethnicity in teachers' categorization of student needs as "exceptional" and in need of special or gifted education services. I use an original survey experiment in which teachers read case studies of fictional male students in which the race/ethnicity, English Language Learner status, and exceptionality characteristics were experimentally manipulated. The teachers are then asked whether they would refer the student for exceptionality testing. My findings suggest a complex intersection of race/ethnicity and exceptionality, in which white boys are more likely to be suspected of having exceptionalities when they exhibit academic challenges, while boys of color are more likely to be suspected when they exhibit behavioral challenges. This suggests that the racialized construction of exceptionalities reflects differential academic expectations and interpretations of behavior by race/ethnicity, with implications for the subjectivity of exceptionality identification and for the exacerbation of racial/ethnic inequalities in education.


Structure and mechanism of an aspartimide-dependent peptide ligase in human legumain.
Peptide ligases expand the repertoire of genetically encoded protein architectures by synthesizing new peptide bonds, energetically driven by ATP or NTPs. Here, we report the discovery of a genuine ligase activity in human legumain (AEP) which has important roles in immunity and tumor progression that were believed to be due to its established cysteine protease activity. Defying dogma, the ligase reaction is independent of the catalytic cysteine but exploits an endogenous energy reservoir that results from the conversion of a conserved aspartate to a metastable aspartimide. Legumain's dual protease-ligase activities are pH- and thus localization controlled, dominating at acidic and neutral pH, respectively. Their relevance includes reversible on-off switching of cystatin inhibitors and enzyme (in)activation, and may affect the generation of three-dimensional MHC epitopes. The aspartate-aspartimide (succinimide) pair represents a new paradigm of coupling endergonic reactions in ATP-scarce environments.


Mycobacterium tuberculosis WhiB4 regulates oxidative stress response to modulate survival and dissemination in vivo.
Host-generated oxidative stress is considered one of the main mechanisms constraining Mycobacterium tuberculosis (Mtb) growth. The redox-sensing mechanisms in Mtb are not completely understood. Here we show that WhiB4 responds to oxygen (O₂) and nitric oxide (NO) via its 4Fe-4S cluster and controls the oxidative stress response in Mtb. The WhiB4 mutant (MtbΔwhiB4) displayed an altered redox balance and a reduced membrane potential. Microarray analysis demonstrated that MtbΔwhiB4 overexpresses the antioxidant systems including alkyl hydroperoxidase (ahpC-ahpD) and rubredoxins (rubA-rubB). DNA binding assays showed that WhiB4 [4Fe-4S] cluster is dispensable for DNA binding. However, oxidation of the apo-WhiB4 Cys thiols induced disulphide-linked oligomerization, DNA binding and transcriptional repression, whereas reduction reversed the effect. Furthermore, WhiB4 binds DNA with a preference for GC-rich sequences. Expression analysis showed that oxidative stress repressed whiB4 and induced antioxidants in Mtb, while their hyper-induction was observed in MtbΔwhiB4. MtbΔwhiB4 showed increased resistance to oxidative stress in vitro and enhanced survival inside the macrophages. Lastly, MtbΔwhiB4 displayed hypervirulence in the lungs of guinea pigs, but showed a defect in dissemination to their spleen. These findings suggest that WhiB4 systematically calibrates the activation of oxidative stress response in Mtb to maintain redox balance, and to modulate virulence.


The relation between language and cognition in 3- to 9-year-olds: the acquisition of grammatical gender in French.
The French language has a grammatical gender system in which all nouns are assigned either a masculine or a feminine gender. Nouns provide two types of gender cues that can potentially guide gender attribution: morphophonological cues carried by endings and semantic cues (natural gender). The first goal of this study was to describe the acquisition of the probabilistic system based on phonological oppositions on word endings by French-speaking children. The second goal was to explore the extent to which this system affects categorization. In the study, 3- to 9-year-olds assigned gender categorization to invented nouns whose endings were typically masculine, typically feminine, or neutral. Two response conditions were used. In the determiner condition, children indicated the gender class by orally providing the determiner un or une marked for gender. In the picture condition, responses were given by pointing to the picture of a Martian-like female or male person that would be best called by each spoken pseudoword. Results indicated that as young as 3 years, children associated the determiner corresponding to the ending bias at greater than chance levels. Ending-consistent performance increased from 3 to 9 years of age. Moreover, from 4 years of age onward, sensitivity to endings affected categorization. Starting at that age, pictures were selected according to endings at greater than chance levels. This effect also increased with age. The discussion deals with the mechanisms of language acquisition and the relation between language and cognition.


Inducible trans-activation of plastid transgenes: expression of the R. eutropha phb operon in transplastomic tobacco.
Deleterious effects of constitutive transgene expression can occur if gene products are harmful to the transformed plant. Constraints such as growth inhibition and male sterility have been observed in plastid transformants containing the phb operon encoding the genes required for the production of the polyester polyhydroxybutyric acid (PHB). In order to induce PHB synthesis in tobacco in a well-timed manner, we have constructed a trans-activation system to regulate transcription of the phb operon in plastids. This system consists of a nuclear-located, ethanol-inducible T7RNA polymerase (T7RNAP) which is targeted to plastids harboring the phb operon under control of T7 regulatory elements. Following treatment with 5% ethanol, moderate induction of PHB synthesis was found. PHB amounts reached 1,383 ppm in dry weight, and an overall background activity of 171 ppm was measured in uninduced tissues. On the transcriptional level, T7RNAP induction was proven and we found that the phb operon is transcribed into at least two mRNAs. Without ethanol induction, development of flowers and fertile seeds was possible. Thus, the main problem of inhibitory transgene expression was solved. Our results show that this inducible trans-activation system could serve as an alternative to constitutive expression of transgenes in the plastome.


Evaluation of the Antimicrobial Activity of a Super Oxidized Solution in Clinical Isolates.
This study evaluated the antimicrobial activity of Estericide(®) QX (super oxidized solution) in 524 bacterial clinical isolates causing nosocomial infections. The minimum inhibitory concentration (MIC) was determined by the serial broth microdilution method. The bacterial viability of the isolates and control strains was tested. The bactericidal effect of the disinfectant was determined according to the European Standards (EN) Test Methods-1040 guidelines. Assay of stability in Estericide QX after 1 year of storage was performed. The microdilution assays showed that the isolates were inhibited at concentrations of 10-40 parts per million (ppm). For gram-positive bacteria, the MIC values 20 and 40 ppm were more predominant (95%), whereas for gram-negative bacteria, the MIC values 10 and 20 ppm had the highest percentage (91.7%). The difference between the two groups was statistically significant (p<0.001). The results of the assay of bactericidal activity showed that all tested bacteria (99.999%) were killed within 30 sec of contact time. The stability test showed that Estericide QX maintained its disinfectant action over time. In conclusion, the results of the present study showed that the super oxidized solution of Estericide QX provides a high antibacterial activity on both gram-positive and gram-negative bacteria. Based on these results and under the conditions of the present study, we believe that Estericide QX can be used efficiently against multiresistant nosocomial bacteria, providing an opportunity for new disinfection alternatives.


Male androphilia in the ancestral environment. An ethnological analysis.
The kin selection hypothesis posits that male androphilia (male sexual attraction to adult males) evolved because androphilic males invest more in kin, thereby enhancing inclusive fitness. Increased kin-directed altruism has been repeatedly documented among a population of transgendered androphilic males, but never among androphilic males in other cultures who adopt gender identities as men. Thus, the kin selection hypothesis may be viable if male androphilia was expressed in the transgendered form in the ancestral past. Using the Standard Cross-Cultural Sample (SCCS), we examined 46 societies in which male androphilia was expressed in the transgendered form (transgendered societies) and 146 comparison societies (non-transgendered societies). We analyzed SCCS variables pertaining to ancestral sociocultural conditions, access to kin, and societal reactions to homosexuality. Our results show that ancestral sociocultural conditions and bilateral and double descent systems were more common in transgendered than in non-transgendered societies. Across the entire sample, descent systems and residence patterns that would presumably facilitate increased access to kin were associated with the presence of ancestral sociocultural conditions. Among transgendered societies, negative societal attitudes toward homosexuality were unlikely. We conclude that the ancestral human sociocultural environment was likely conducive to the expression of the transgendered form of male androphilia. Descent systems, residence patterns, and societal reactions to homosexuality likely facilitated investments in kin by transgendered males. Given that contemporary transgendered male androphiles appear to exhibit elevated kin-directed altruism, these findings further indicate the viability of the kin selection hypothesis.


Induction of autophagy and cell death by tamoxifen in cultured retinal pigment epithelial and photoreceptor cells.
We investigated the mechanism of tamoxifen (TAM) retinotoxicity using human retinal pigment epithelial (RPE)-derived (ARPE-19) and photoreceptor-derived (661W) cells. Cultured ARPE-19 and 661W cells were treated with 5 to 10 μM TAM, and the resultant cell death was quantified using lactate dehydrogenase (LDH) release assay. Cellular oxidative stress was determined by measuring 5-(and-6)-carboxy-2',7'-dichlorohydrofluorescein diacetate (H(2)-DCFDA) fluorescence. Changes in intracellular free zinc levels were monitored using the zinc-specific fluorescent dye, FluoZin-3 AM. Autophagic vacuole formation was assessed morphologically in ARPE-19 and 661W cells transfected with the fluorescent protein-conjugated markers, RFP-LC3 or GFP-LC3. Following exposure to TAM, both ARPE-19 and 661W cells had cytosolic vacuoles within 1 hour and underwent cell death within 18 hours. In both cell types, TAM-induced cell death was accompanied by increased oxidative stress and elevated zinc levels, and was attenuated by the antioxidant N-acetyl-L-cysteine (NAC) or the zinc chelator N,N,N'N'-tetrakis(-)(2-pyridylmethyl)-ethylenediamine (TPEN). The levels of LC3-II as well as the number of autophagic vacuoles (AVs) increased after TAM treatment. Double staining for lysosomes and AVs showed that autolysosome formation proceeded normally. Consistent with this, autophagy flux was increased. Finally, as shown in other cases of autophagic cell death, lysosomal membrane permeabilization (LMP) as well as caspase-dependent apoptosis contributed to TAM-induced cell death. ARPE-19 and 661W cells were vulnerable similarly to TAM-induced cytotoxicity. Increases in zinc levels and oxidative stress, excessive activation of autophagy flux, and ultimately the occurrence of LMP and consequent caspase activation may contribute to the well-established retinal cytotoxicity of TAM.


COMMD1-mediated ubiquitination regulates CFTR trafficking.
The CFTR (cystic fibrosis transmembrane conductance regulator) protein is a large polytopic protein whose biogenesis is inefficient. To better understand the regulation of CFTR processing and trafficking, we conducted a genetic screen that identified COMMD1 as a new CFTR partner. COMMD1 is a protein associated with multiple cellular pathways, including the regulation of hepatic copper excretion, sodium uptake through interaction with ENaC (epithelial sodium channel) and NF-kappaB signaling. In this study, we show that COMMD1 interacts with CFTR in cells expressing both proteins endogenously. This interaction promotes CFTR cell surface expression as assessed by biotinylation experiments in heterologously expressing cells through regulation of CFTR ubiquitination. In summary, our data demonstrate that CFTR is protected from ubiquitination by COMMD1, which sustains CFTR expression at the plasma membrane. Thus, increasing COMMD1 expression may provide an approach to simultaneously inhibit ENaC absorption and enhance CFTR trafficking, two major issues in cystic fibrosis.


Prevalence, incidence and risk factors of strabismus in a Chinese population-based cohort of preschool children: the Nanjing Eye Study.
To evaluate the prevalence, incidence and their related risk factors of strabismus among preschool children in China. Children born between September 2011 and August 2012 in Yuhuatai District of Nanjing were invited to participate in the Nanjing Eye Study for a comprehensive eye examination annually since 2015. The data presented in this paper were obtained from 2015 to 2017, when these children grew from the age of 3 to 5 years. Eye examinations included visual acuity, anterior segment, posterior segment, refraction, and ocular alignment and motility. Risk factors were evaluated using univariable and multivariable logistic regression models for prevalent and incident strabismus. In 2015, a total of 2018 children (87.7% response rate) of 2300 eligible preschoolers completed the baseline eye examination when they were 3 years old. Among the 2018 participants, 50 had strabismus (prevalence rate, 2.48%). In multivariable analysis, prevalent strabismus was independently associated with parental strabismus history (OR=11.60, p<0.001), hyperopia (OR=6.22, p<0.001), prematurity (OR=3.07, p=0.01) and astigmatism (OR=2.15, p=0.04). Among 1766 children followed up for 2 years, 63 developed strabismus (annual incidence rate, 1.78%), of whom 57 had exotropia and 6 had esotropia. In multivariable analysis, incident strabismus was significantly associated with parental strabismus history (OR=5.55, p=0.04) and prematurity (OR=3.77, p<0.001). In this population-based cohort study, we found a higher incidence of strabismus and a higher exotropia:esotropia ratio than previous studies in preschool children. Parental strabismus history and prematurity were associated with a higher risk for both prevalent and incident strabismus.


Mechanism of toxicity in rotenone models of Parkinson's disease.
Exposure of rats to the pesticide and complex I inhibitor rotenone reproduces features of Parkinson's disease, including selective nigrostriatal dopaminergic degeneration and alpha-synuclein-positive cytoplasmic inclusions (Betarbet et al., 2000; Sherer et al., 2003). Here, we examined mechanisms of rotenone toxicity using three model systems. In SK-N-MC human neuroblastoma cells, rotenone (10 nm to 1 microm) caused dose-dependent ATP depletion, oxidative damage, and death. To determine the molecular site of action of rotenone, cells were transfected with the rotenone-insensitive single-subunit NADH dehydrogenase of Saccharomyces cerevisiae (NDI1), which incorporates into the mammalian ETC and acts as a "replacement" for endogenous complex I. In response to rotenone, NDI1-transfected cells did not show mitochondrial impairment, oxidative damage, or death, demonstrating that these effects of rotenone were caused by specific interactions at complex I. Although rotenone caused modest ATP depletion, equivalent ATP loss induced by 2-deoxyglucose was without toxicity, arguing that bioenergetic defects were not responsible for cell death. In contrast, reducing oxidative damage with antioxidants, or by NDI1 transfection, blocked cell death. To determine the relevance of rotenone-induced oxidative damage to dopaminergic neuronal death, we used a chronic midbrain slice culture model. In this system, rotenone caused oxidative damage and dopaminergic neuronal loss, effects blocked by alpha-tocopherol. Finally, brains from rotenone-treated animals demonstrated oxidative damage, most notably in midbrain and olfactory bulb, dopaminergic regions affected by Parkinson's disease. These results, using three models of increasing complexity, demonstrate the involvement of oxidative damage in rotenone toxicity and support the evaluation of antioxidant therapies for Parkinson's disease.


The entire beta-globin gene cluster is deleted in a form of gamma delta beta-thalassemia.
We have used restriction endonuclease mapping to study a deletion involving the beta-globin gene cluster in a Mexican-American family with gamma delta beta-thalassemia. Analysis of DNA polymorphisms demonstrated deletion of the beta-globin gene from the affected chromosome. Using a DNA fragment that maps greater than 40 kilobases (kb) 5' to the epsilon-gene as a probe, reduced amounts of normal fragments were found in the DNA of affected family members. Similar analysis using radiolabeled DNA fragments located 3' to the beta-globin cluster has shown that the deletion extends more than 17 kb 3' to the beta-gene, but terminates before the 3' endpoint of the Ghanian HPFH deletion. Hence, this gamma delta beta-thalassemia deletion eliminates over 105 kb of DNA and is the first report of a deletion of the entire beta-globin gene cluster.


The phylogeny of closely related species as revealed by the genealogy of a speciation gene, Odysseus.
Molecular differentiation between races or closely related species is often incongruent with the reproductive divergence of the taxa of interest. Shared ancient polymorphism and/or introgression during secondary contact may be responsible for the incongruence. At loci contributing to speciation, these two complications should be minimized (1, 2); hence, their variation may more faithfully reflect the history of the species' reproductive differentiation. In this study, we analyzed DNA polymorphism at the Odysseus (OdsH) locus of hybrid sterility between Drosophila mauritiana and Drosophila simulans and were able to verify such a prediction. Interestingly, DNA variation only a short distance away (1.8 kb) appears not to be influenced by the forces that shape the recent evolution of the OdsH coding region. This locus thus may represent a test case of inferring phylogeny of very closely related species.


[Gangliocytic paraganglioma of the ampulla of Vater].
A case of a bleeding duodenal gangliocytic paraganglioma (GPG) of the Papilla Vateri is reported. The 47 year old patient was treated successfully by partial pancreaticoduodenectomy. Histologically the GPG is characterized by three types of cells: epitheloid cells, ganglion-like cells and spindle cells. So far 81 cases are reported in the literature. Most cases of GPGs are benign, metastasis or recurrence is possible. This case showed an infiltrative growth pattern indicating the true neoplastic nature of this tumor.


A selective peroxisome proliferator-activated receptor delta agonist promotes reverse cholesterol transport.
The peroxisome proliferator-activated receptors (PPARs) are dietary lipid sensors that regulate fatty acid and carbohydrate metabolism. The hypolipidemic effects of the fibrate drugs and the antidiabetic effects of the glitazone drugs in humans are due to activation of the alpha (NR1C1) and gamma (NR1C3) subtypes, respectively. By contrast, the therapeutic potential of the delta (NR1C2) subtype is unknown, due in part to the lack of selective ligands. We have used combinatorial chemistry and structure-based drug design to develop a potent and subtype-selective PPARdelta agonist, GW501516. In macrophages, fibroblasts, and intestinal cells, GW501516 increases expression of the reverse cholesterol transporter ATP-binding cassette A1 and induces apolipoprotein A1-specific cholesterol efflux. When dosed to insulin-resistant middle-aged obese rhesus monkeys, GW501516 causes a dramatic dose-dependent rise in serum high density lipoprotein cholesterol while lowering the levels of small-dense low density lipoprotein, fasting triglycerides, and fasting insulin. Our results suggest that PPARdelta agonists may be effective drugs to increase reverse cholesterol transport and decrease cardiovascular disease associated with the metabolic syndrome X.


A lamin-dependent pathway that regulates nuclear organization, cell cycle progression and germ cell development.
The C. elegans genome encodes a single lamin protein (Ce-lamin), three LEM domain proteins (Ce-emerin, Ce-MAN1 and LEM-3) and a single BAF protein (Ce-BAF). Down-regulation of Ce-lamin causes embryonic lethality. Abnormalities include rapid changes in nuclear morphology during interphase, inability of cells to complete mitosis, abnormal condensation of chromatin, clustering of nuclear pore complexes (NPCs), and missing or abnormal germ cells. Ce-emerin and Ce-MAN1 are both embedded in the inner nuclear membrane, and both bind Ce-lamin and Ce-BAF; in addition, both require Ce-lamin for their localization. Mutations in human emerin cause X-linked recessive Emery-Dreifuss muscular dystrophy. In C. elegans, loss of Ce-emerin alone has no detectable phenotype, while loss of 90% Ce-MAN1 causes approximately 15% embryonic lethality. However in worms that lack Ce-emerin, a approximately 90% reduction of Ce-MAN1 is lethal to all embryos by the 100-cell stage, with a phenotype involving chromatin condensation and repeated cycles of anaphase chromosome bridging and cytokinesis. The anaphase-bridged chromatin retained a mitosis-specific phosphohistone H3 epitope, and failed to recruit detectable Ce-lamin or Ce-BAF. Down-regulation of Ce-BAF showed similar phenotypes. These findings suggest that lamin, LEM-domain proteins and BAF are part of a lamina network essential for chromatin organization and cell division, and that Ce-emerin and Ce-MAN1 share at least one and possibly multiple overlapping functions, which may be relevant to Emery-Dreifuss muscular dystrophy.


Curcumin as a Therapeutic Strategy in Liver Diseases.
Liver diseases are classified as acute and chronic hepatic failures. In particular, chronic pathologies are the most common diseases in the World. Chronic pathologies of liver disease are the most common diseases in the world. There are many causes that induce a progressive and irreversible degeneration of the hepatic parenchyma, but, in general, they lead to the destruction of the normal balance between reactive oxygen stress (ROS) formation and ROS degradation within the liver. The prevalence of disabling diseases, including the hepatic diseases, is increasingly widespread, and it is important to find a safe, inexpensive, accessible and effective way to face this condition. Many recent studies have focused on different natural antioxidants, which could restore the physiological hepatic environment, thereby allowing the normal functioning of this organ. Natural products have been used to discover new leads for treating several diseases; among them, it is important to emphasize curcumin, which is a polyphenol obtained from Curcuma longa Linn, a plant naturally found throughout tropical and subtropical regions of the world.


Pre-scaled two-parameter Gauss-Newton image reconstruction to reduce property recovery imbalance.
Gauss-Newton image reconstruction in microwave imaging can be formulated in terms of a single complex quantity, the wave number squared (k2), with the understanding that the relative permittivity and conductivity images can be extracted afterwards through a simple constitutive relationship. However, this approach ignores the fact that the magnitude of the average real and imaginary components can be considerably out of balance depending on the operating frequency and tissue characteristics which can inadvertently imbalance the process in favour of one parameter over the other. In an effort to achieve property recovery which is balanced, we introduce a pre-scaling procedure at the property update stage of the reconstruction. Utilization of this concept in conjunction with our two-step regularization process for both simulation and phantom experiments demonstrates that the penalty term weighting parameters for the optimal mean-squared property errors for the two recovered distributions (relative permittivity and conductivity) together with that yielding the lowest least-squared electric field error coincide only when the scaling is applied. The scheme provides a means for simultaneous optimization of the two permittivity and conductivity images.


Comparison of prognosis after enucleation vs cobalt 60 irradiation of melanomas.
Between December 1962 and May 1979, growth of choroidal and ciliary body melanomas was documented in 34 patients prior to enucleation. Twenty-seven of the 34 patients underwent enucleation during the same time period (1967 through 1976), when primary treatment with a cobalt 60 episcleral plaque was employed in a nonrandomized fashion in 21 patients with similar-sized melanomas. Comparison of these two groups of patients, none of whom was unavailable for follow-up and all of whom were followed up for a minimum of 66 months, showed that the probability of dying of metastatic melanoma within five years after cobalt 60 therapy was 50% compared with 16% after enucleation. Factors of potential importance in explaining this difference included nonrandomization, small number of patients in both groups, and greater frequency of anterior location of tumors in the cobalt 60 group (76% vs 22%). A randomized, controlled clinical trial is required to determine the relative safety of treatment for uveal melanomas.


Lithium's effect of parathyroid hormone.
Lithium has been reported to raise serum calcium and lower serum phosphate concentrations and to increase urinary calcium excretion. Because these changes may be effects of parathyroid hormone (PTH), PTH was measured in 19 patients receiving lithium. PTH was significantly higher in these patients than in 150 normal subjects. For all patients serum calcium concentrations correlated significantly with serum lithium concentrations. These results indicate that lithium may cause biochemical hyperparathyroidism. Secondary hyperparathyroidism in certain patients with lithium nephrotoxicity is also possible.