Demographic, clinical and behavioural determinants of HIV serostatus non-disclosure to sex partners among HIV-infected pregnant women in the Eastern Cape, South Africa.
Drawing from a baseline sample of a cohort study, the study examines the extent and correlates of serostatus non-disclosure to sex partners and family members, and reasons for non-disclosure among HIV-infected pregnant women in the Eastern Cape Province, South Africa. This longitudinal cohort study recruited 1709 pregnant women living with HIV who attended three of the largest maternity centres in the Eastern Cape, South Africa, for delivery between September 2015 and May 2016. Relevant items on demographics, serostatus awareness, disclosure to sex partners and family members, and lifestyle behaviours were obtained using structured interviews. Age-stratified binary logistic regression models were used to determine the significant correlates of non-disclosure among the participants. A higher rate of HIV serostatus non-disclosure to sex partners (25.6%) in comparison to family members (20%) was reported by the participants. Younger age, not living with partners and alcohol use were significantly associated with non-disclosure of HIV serostatus to sex partners. Non-disclosure of HIV serostatus to sex partners was significantly (p<0.05) associated with poor adherence to the highly active anti-retroviral therapy (HAART), failure to keep clinic appointments and high viral load at the delivery of the baby. Perceived fear of intimate partner violence, fear of rejection, guilt of not disclosing at the onset of the relationship, sex partner's non-disclosure of HIV serostatus, and guilt of unfaithfulness were some of the reasons for non-disclosure of HIV serostatus to sex partners. Non-disclosure of HIV serostatus is a public health concern with serious implications for both mother-to-child transmission, as well as horizontal transmission, in our setting. Strategic efforts toward ending the epidemic of HIV and AIDS in South Africa should address the sociocultural and behavioural determinants of non-disclosure.


Synthesis, cytotoxicity, DNA interaction and cell cycle studies of trans-diiodophosphine Pt(II) complexes.
Platinum complexes, bearing aliphatic amines and phosphine ligands in trans configuration with iodide as leaving groups, are synthesized and characterized. The crystal structure of trans-PtI2(isopropylamine)(PPh3) is reported. The complex bearing isopropylamine is demonstrated to be the best candidate as its cytotoxic activity is comparable to or better than cisplatin. A remarkably higher interaction of the complexes with DNA is reported as compared to the parent chlorido series. Cell cycle studies of the complexes in six human cell lines are performed and also compared with the previous series.


IFPA meeting 2015 workshop report III: nanomedicine applications and exosome biology, xenobiotics and endocrine disruptors and pregnancy, and lipid.
Workshops are an important part of the IFPA annual meeting, as they allow for discussion of specialized topics. At the IFPA meeting 2015 there were twelve themed workshops, three of which are summarized in this report. These workshops were related to various aspects of placental biology but collectively covered areas of pregnancy pathologies and placental metabolism: 1) nanomedicine applications and exosome biology; 2) xenobiotics and endocrine disruptors and pregnancy; 3) lipid mediators and placental function.


An Algorithm for Strategic Continuation or Restriction of Asthma Medication Prior to Exercise Challenge Testing in Childhood Exercise Induced Bronchoconstriction.
Exercise induced bronchial (EIB) constriction is a common and highly specific feature of pediatric asthma and should be diagnosed with an exercise challenge test (ECT). The impact of EIB in asthmatic children's daily lives is immense, considering the effects on both physical and psychosocial development. Monitoring childhood asthma by ECT's can provide insight into daily life disease burden and the control of asthma. Current guidelines for bronchoprovocation tests restrict both the use of reliever and maintenance asthma medication before an exercise challenge to prevent false-negative testing, as both have significant acute bronchoprotective properties. However, restricting maintenance medication before an ECT may be less appropiate to evaluate EIB symptoms in daily life when a diagnosis of asthma is well established. Rigorous of maintenance medication before an ECT according to guidelines may lead to overestimation of the real, daily life asthma burden and lead to an inappropiate step-up in therapy. The protection against EIB offered by the combined acute and chronic bronchoprotective effects of maintenance medication can be properly assessed whilst maintaining them. This may aid in achieving the goal of unrestricted participation of children in daily play and sports activities with their peers without escalation of therapy. When considering a step down in medication, a strategic wash-out of maintenance medication before an ECT aids in providing objective support of potential discontinuation of maintenance medication.


A clinical study to evaluate the usefulness of the MAST test in diagnosing pre-labour rupture of membranes.
An open study was performed to evaluate the efficiency of insulin-like growth factor binding protein-1 as a marker for the detection of pre-labour rupture of membranes and to assess whether it aided the subsequent clinical management. One hundred women between 24 and 42 weeks' gestation with a history suggestive of pre-labour rupture of membranes were studied. There were 11 false positive and no false negative results and the sensitivity and specificity were 100% and 89% respectively. The presence of infection, blood or semen did not interfere with the accuracy of the results.


Evaluation of Third-Party Deep Cleaning as a Listeria monocytogenes Control Strategy in Retail Delis.
The objective of this study was to develop and assess the efficacy of an aggressive deep cleaning sanitation standard operating procedure (DC-SSOP) in nine retail delicatessens to reduce persistent Listeria monocytogenes environmental contamination. The DC-SSOP was developed from combined daily SSOPs recommended by the Food Marketing Institute and input from experts in Listeria control from food manufacturing and sanitation. The DC-SSOP was executed by a trained professional cleaning service during a single 12-h shutdown period. A modified protocol from the U.S. Food and Drug Administration Bacteriological Analytical Manual was used to detect L. monocytogenes in samples from 28 food and nonfood contact surfaces that were collected immediately before and after each cleaning and in samples collected monthly for 3 months. The DC-SSOP significantly reduced L. monocytogenes prevalence overall during the 3-month follow-up period and produced variable results for persistent L. monocytogenes isolates. Six delis with historically low to moderate L. monocytogenes prevalence had no significant changes in the number of samples positive for L. monocytogenes after deep cleaning. Deep cleaning in very high prevalence delis (20 to 30% prevalence) reduced L. monocytogenes by 25.6% (Padj < 0.0001, n = 294) overall during the follow-up period. Among delis with extremely high prevalence (>30%), positive samples from nonfood contact surfaces were reduced by 19.6% (Padj = 0.0002, n = 294) during the follow-up period. The inability of deep cleaning to completely eliminate persistent L. monocytogenes was likely due to the diverse infrastructures in each deli, which may require more individualized intervention strategies.


Para-Bombay Phenotype of a Pregnant Mother in Malaysia: Transfusion for an Extremely Premature Baby.
Para-Bombay blood phenotype is a rare blood group with limited cases reported worldwide. This blood group is characterized by the absence of ABH antigen on red blood cells but presence of ABH secretor substances in the body secretion. This rare phenotype is usually misinterpreted as O and may endanger the patient if urgent blood transfusion is required. A mother who was labelled as group O Rh D positive during antenatal follow-up was found to have ABO discrepancy during delivery. The newborn was admitted for extremely premature delivery at 25 weeks. As the baby required transfusion, problem arose during cross matching with the mother's sample. It was found that the mother was group O Rh D positive in forward grouping. However, the reverse grouping showed the presence of reaction (2+) in O cells. The baby was grouped as O Rh D positive. As transfusion was urgently needed due to baby's unstable condition, group O Rh D positive packed cell was found compatible with baby's serum, subsequently transfused. Bombay blood donor was contacted, and the donated blood was sent to the hospital for further management. Further investigations were performed, indicating that the mother is para-Bombay A. Due to recent transfusion to baby, we suggested to repeat baby's blood group after the baby is one year old. Para-Bombay was usually mislabelled as O if the sample was not tested with O cell in reverse grouping. Additional tests may be needed during antenatal follow-up to prevent complications during delivery, which requires emergency blood transfusion.


A close look at autoimmune muscle disorders: association of Lambert-Eaton myasthenic syndrome with dermatomyositis.
Dermatomyositis/polymyositis (DM/PM) and Lambert-Eaton myasthenic syndrome (LEMS) are two autoimmune disorders that have very rarely been reported to occur together in the same patient. We report on two patients with DM who were later diagnosed with concomitant LEMS, and point out diagnostic challenges in identifying LEMS in patients with DM/PM. As specific treatment for LEMS is available, it is important to identify those DM/PM patients who suffer from concomitant LEMS.


Viability of the isolated soleus muscle during long-term incubation.
Skeletal muscle metabolism has been examined in perfused hindlimb muscles and in isolated muscle preparations. While long-term viability of the fast-twitch epitrochlearis has been documented with respect to glucose transport, it appears that long-term incubated soleus muscles are less stable when incubated ex vivo for many hours. Therefore, in the present study, we have examined whether the isolated soleus muscle remains metabolically viable for up to 18 h with respect to maintaining ATP and phosphocreatine (PCr) concentrations, carbohydrate and fatty-acid metabolism, insulin signalling, and protein expression. Soleus muscles were incubated in well-oxygenated Medium 199 (M199) supplemented with low concentrations of insulin (14.3 microU/mL) for 0, 6, 12, and 18 h. During this incubating period the concentrations of ATP and PCr were stable, indicating that oxygenation and substrate supply were being maintained. In addition, the concentrations of proglycogen and macroglycogen were not altered, whereas an increase (+30%) in intramuscular triacylglycerol concentration was observed at the end of 18 h of incubation (p < 0.05). Complex molecular processes in the long-term incubated muscles were also stable. This was shown by maintenance of basal as well as insulin-stimulated rates of 3-O-methyl glucose transport, and by the maintenance of protein expression of the glucose transporter GLUT4 and the fatty acid transporters FAT/CD36 and FABPpm. In addition, the insulin-stimulated translocation of GLUT4 to the plasma membrane, which involves a complex signalling cascade, was fully preserved. In conclusion, in well-oxygenated soleus muscles maintained in M199 supplemented with extremely low concentrations of insulin, ATP and PCr concentrations, carbohydrate and fatty acid metabolism, insulin signalling, and protein expression were stably maintained for up to 18 h. This provides for opportunities to examine muscle metabolic function under very highly controlled conditions.


The evaluation of hospice home care volunteers.
A national sample of 139 hospice volunteer coordinators rated a series of statements about volunteer behaviors in terms of their importance to volunteer performance. Each statement had to be rated at least 8 on a scale of 10 by 75% of the respondents in order to be included in the final instrument. This procedure yielded a scale of 27 descriptors of appropriate volunteer behavior. The scale is intended for use by hospice volunteer coordinators who want a tool with which to offer concrete, behavior based, constructive feedback to volunteers about their performance.


Sr-Nd-Hf isotopic analysis of reference materials and natural and anthropogenic particulate matter sources: Implications for accurately tracing North African dust in complex urban atmospheres.
We present novel chemical separation protocols for isotopic analysis of low mass aliquots (0.3 mg and 25 mg) of several reference materials and real-world samples of relevance to urban airborne particulate matter (PM) investigations. A high-yielding gravity flow column chromatography scheme was developed for facile and quantitative separation of Sr, Nd, and Hf prior to multi collector - inductively coupled plasma - mass spectrometry (MC-ICP-MS). Because we are interested in isolating and accurately quantitating individual anthropogenic and natural aerosol sources in complex industrial/metropolitan atmospheric environments, laboratory protocols were optimized using National Institute of Standards and Technology Standard Reference Material (SRM) 1648a (urban atmospheric PM), SRM 1633b (coal fly ash), and European Commission standards BCR-723 (vehicular road dust), and BCR-2 (basalt rock standard). Sr, Nd, and Hf procedural blanks from column chromatography were low (averaging only 37 pg, 17 pg, 11 pg, respectively) and recoveries were high (averaging 95%, 82%, and 92%, respectively). A volume-adjustment protocol was established using isotope reference solutions SRM 987 (SrCO3), JNdi (Nd2O3), and in-house Hf standards to dilute the dried samples prior to MC-ICP-MS based on projected uncertainties for low sample masses. 87Sr/86Sr, 143Nd/144Nd, and 176Hf/177Hf isotopic ratios in SRM 1648a, BCR-723, and SRM 1633b are reported for the first time that can serve as provisional reference values. The novel method was used to characterize isotopic ratios and elemental abundances in two anthropogenic urban aerosol sources, namely motor vehicles and petroleum refining using airborne fine PM collected in a vehicular tunnel and fluidized-bed catalytic cracking catalysts, respectively. Two other important mineral-rich urban PM sources, namely soil (i.e., resuspended crustal material) and concrete/cement dust (i.e., construction activity) were also characterized. These are the first isotopic measurements in these environmental compartments and were compared with literature data for long-range transported North African dust, which is a prominent summertime PM source in urban regions in southeastern United States. We demonstrate the capability of coupled Sr-Nd-Hf isotopes to uniquely trace different mineral dust sources with overlapping elemental composition (Sahara-Sahel region, local soil, and concrete/cement) and accurately isolate various urban PM sources demonstrating the superiority of isotopic markers over elemental tracers.


Does emancipation devour its children? Beyond a stalled dialectic of emancipation.
Emancipation serves not only as a midwife for progressive agendas such as greater equality and sustainability but also as their gravedigger. This diagnosis underpins Ingolfur Blühdorn's 'dialectic of emancipation', which depicts a dilemma but offers no perspective on how to deal with it. By drawing on Foucault, this article suggests conceiving of emancipation as a task moderns are confronted with even if a given emancipatory project has come to devour its children. Claiming autonomy from given social constellations is key to this task; key also is judging between legitimate and illegitimate claims to autonomy. In late modernity, the criteria for such judgement are no longer universally given. Instead of regarding the latter as entry into mere subjectivism (Blühdorn), this article presents judgement as a key political, 'world building'-activity (Arendt), a critical social theory may join in, by not only observing the world but by also taking sides in it.


D-Amino acid substituted peptides as potential alternatives of homochiral L-configurations.
On the primitive Earth, both L- and D-amino acids would have been present. However, only L-amino acids are essential blocks to construct proteins in modern life. To study the relative stability of D-amino acid substituted peptides, a variety of computational methods were applied. Ten prebiotic amino acids (Gly, Ala, Asp, Glu, Ile, Leu, Pro, Ser, Thr, and Val) were previously determined by multiple meteorite, spark discharge, and hydrothermal vent studies. Some previously reported early Earth polypeptide analogs were focused on in this study. Tripeptides composed of only Asp, Ser, and Val exemplified that different positions (i.e., N-terminus, C-terminus, and middle) made a difference in the minimal folding energy of peptides, while the chemical classification of amino acid (hydrophobic, acidic, or hydroxylic) did not show a significant difference. Hierarchical cluster analysis for dipeptides with all possible combinations of the proposed ten prebiotic amino acids and their D-amino acid substituted derivatives generated five clusters. Primordial simple polypeptides were modeled to test the significance of molecular fluctuations, secondary structure occupancies, and folding energy differences based on these clusters. We found peptides with α-helices, long β-sheets, and long loops are usually less sensitive to D-amino acid replacements in comparison to short β-sheets. Intriguingly, amongst 129 D-amino acid residues, mutation sensitivity profiles presented that the ratio of more to less stable residues was about 1. In conclusion, some combinations of a mixture of L- and D-amino acids can potentially act as essential building blocks of life.


Quantification of valvular regurgitation using radioisotopes.
Mitral and aortic regurgitations impose an abnormal volume overload on the left ventricle. Recent advances in radionuclide angiographic measurements of all cardiac volumes have made this a practical technique for the detection, quantification and functional assessment of valvular regurgitations and shunts. The method is based on the comparative evaluation of total and effective left ventricular stroke volume. In the radionuclide technique, the right ventricular stroke volume is most frequently used to represent the effective left ventricular stroke volume although techniques have been presented which used as reference the left ventricular stroke volume calculated from measurements of heart rate and cardiac output (Fick method or dye dilution or scintigraphic techniques). The technique can be performed either during first-pass or at equilibrium. Equilibrium measurements are performed in the left anterior oblique position. The stroke volume ratio and the regurgitant fraction are calculated. This technique has been shown to provide adequate quantitative measurements of mitral and aortic regurgitations. Its specificity is adequate with careful positioning and if regions of interest are determined and care is taken to exclude inadequate studies (as these can be prospectively recognized). The technique can separate moderate from severe regurgitation, provide follow-up values for both left ventricular volume and regurgitant fraction, and assess the effect of interventions on the amount of regurgitation. The technique is, however, not adequate to detect mild or minimal regurgitation. In conclusion, equilibrium scintigraphic measurement of valvular regurgitation is an attractive new technique for measuring valvular regurgitation. Its clinical value lies in its simplicity, its reproducibility and its wide applicability. Its accuracy will be improved by performance of gated tomographic acquisitions.


[Can dental equipment interfere with correct functioning of an ICD?]
An implantable cardioverter defibrillator (ICD) is often placed in patients at high risk of ventricular tachycardia and ventricular fibrillation. An ICD is potentially sensitive to electromagnetic interference (EMI) from dental equipment. Between 2000 to 2020, this was investigated in 7 in vitro studies and 4 in vivo studies. Most dental equipment appears to be safe for routine use in patients with an ICD. However, caution seems appropriate with regard to the use of piezoelectic and magnetostrictive scalers as some can cause EMI up to a distance of 20 to 45 centimeter. Use of an electrotome appears to be contraindicated in ICD wearers.


[Use of ceramic materials in fixed prosthetic appliances].
The development of the use of ceramic materials in stomatology has a long history. Their composition and processing should present in particular improve their strength and accuracy, while preserving excellent biocompatibility and satisfactory aesthetic appearance. Ceramic materials are increasingly used in fixed prosthetic appliances. The author summarizes hitherto assembled findings and outlines the trends of future development.


Associations Between Children's Media Use and Language and Literacy Skills.
Media use is a pervasive aspect of children's home experiences but is often not considered in studies of the home learning environment. Media use could be detrimental to children's language and literacy skills because it may displace other literacy-enhancing activities like shared reading and decrease the quantity and quality of caregiver-child interaction. Thus, the current study asked whether media use is associated with gains in children's language and literacy skills both at a single time point and across a school year and whether age moderates any association. Children (N = 1583) were from preschool through third grade classrooms and language and literacy skills were measured in the fall and spring of the school year. Parents reported how much time their child spends using media on a typical school day. Regression analyses showed that using 4 h or more of media was related to lower literacy gains, but not to language gains. Multilevel models conducted as a robustness check showed that this effect did not hold when accounting for classroom. In neither set of models was there an interaction between age and media use. Single-time-point models did show some associations that did not manifest in more stringent models, highlighting the limitations of correlational designs that do not have measures of children's skills over time. Given the concern and popular press coverage around children's media use, it is important to acknowledge non-significant effects in this domain. These non-significant associations suggest that societal fears around children's media use may be exaggerated. Notably, however, characteristics of children's media use, like educational content or adult co-use, may moderate any effects. The relation between media use and language and literacy growth did not differ across the age range investigated suggesting that, within this range, younger children are not more vulnerable to detrimental effects.


bcr gene rearrangement analysis in myeloproliferative disorders other than chronic myelogenous leukemia.
The bcr gene rearrangement resulting from the Philadelphia translocation is diagnostic of chronic myelogenous leukemia (CML) and is considered the hallmark of this myeloproliferative disorder (MPD) at the molecular level. The other MPDs, essential thrombocythemia (ET), polycythemia vera (PV), agnogenic myeloid metaplasia (AMM), and unclassified MPD, share morphologic features with CML, making the diagnosis difficult in cases with considerable morphologic overlap. In such cases, molecular analysis becomes essential for accurate diagnosis. We report results of bcr analysis by Southern hybridization in 37 patients with MPDs other than CML: ET (20 cases), PV (seven cases). unclassified MPD (nine cases), as well as in one case of chronic myelomonocytic leukemia (CMML). bcr negativity ruled out CML in 36 cases, confirming the morphologic diagnosis. In one case diagnosed as ET. bcr gene rearrangement was diagnostic of CML. The correct diagnosis made possible a different therapeutic approach in this young patient and resulted in cure after allogeneic bone marrow transplantation. The existence of such cases makes the use of molecular analysis essential in the evaluation of MPDs, even when the morphologic features do not unequivocally support the diagnosis of CML, as in this patient.


LncRNA Xist Contributes to Endogenous Neurological Repair After Chronic Compressive Spinal Cord Injury by Promoting Angiogenesis Through the miR-32-5p/Notch-1 Axis.
Endogenous repair after chronic compressive spinal cord injury (CCSCI) is of great clinical interest. Ischemia-hypoxia-induced angiogenesis has been proposed to play an important role during this repair process. Emerging evidence indicates that long non-coding RNAs (lncRNAs) are involved in the pathophysiological processes of various diseases. Here, we identified a lncRNA (Xist; X-inactive specific transcript) with upregulated expression in cervical spine lesions during endogenous neurological repair in CCSCI rats. Therapeutically, the introduction of Xist to rats increased neurological function in vivo as assayed using the Basso, Beattie, and Bresnahan (BBB) score and inclined plane test (IPT). We found that the introduction of Xist enhanced endogenous neurological repair by promoting angiogenesis and microvessel density after CCSCI, while depletion of Xist inhibited angiogenesis and cell sprouting and migration. Mechanistically, Xist promoted angiogenesis by sponging miR-32-5p and modulating Notch-1 expression both in vitro and in vivo. These findings suggest a role of the Xist/miR-32-5p/Notch-1 axis in endogenous repair and provide a potential molecular target for the treatment of ischemia-related central nervous system (CNS) diseases.


The relation of premorbid factors to aggressive physical behavior in dementia.
Aggressive physical behaviour (APB) is common in persons with dementia and often leads to negative consequences such as use of restraints and staff member burnout. For the past several years, a group of nurse researchers has collaborated to develop a model that views dementia behaviors as need-driven but dementia-compromised. The model posits that background variables of the demented person interact with proximal (or current situational) variables to produce APB. The purpose of this study was to test a part of that model by addressing the question: Which premorbid factors predict APB in a sample of 84 demented institutionalized elders? This was a cross-sectional descriptive study that obtained measures of the following characteristics of residents: (1) aggressive behavior as assessed by nursing home staff members using the Ryden Aggression Scale, (2) premorbid personality traits as assessed by a member of the resident's family using the NEO Five Factor Inventory and (3) history of psychosocial stress as assessed by a member of the residents' family using the modified Social Readjustment Rating Scale. Of the sample of 84 residents, 44% exhibited physical aggression. Background factors in the model were not predictive of aggressive behavior in late-stage dementia, although the relation between premorbid neuroticism and physical aggression was in the predicted direction.


Pharmacokinetics of itraconazole following oral administration to normal volunteers.
The pharmacokinetics of itraconazole, an orally effective, broad-spectrum, systemic antifungal agent, were evaluated in five healthy male volunteers. Each subject was studied on days 1 and 15 at the following dosages: 100 mg once daily (regimen A), 200 mg once daily (regimen B), and 200 mg twice daily (regimen C). On each study day, itraconazole was administered with a standardized meal. Plasma samples were collected for 72 h postdose, and 24-h urine specimens were obtained. On day 1 of regimen C, plasma samples were collected following the second dose. Samples were assayed for itraconazole by a sensitive, reverse-phase, high-performance liquid chromatography method. Wide intersubject variations in itraconazole concentration in plasma versus time profiles were observed on all study days. Absorption appeared to be slow, with day 1 mean peak itraconazole concentrations in plasma of 110 ng/ml at 2.8 h (regimen A), 272 ng/ml at 3.0 h (regimen B), and 553 ng/ml at 3.4 h (regimen C). Mean peak itraconazole concentrations in plasma on day 15 were 412 ng/ml at 3.0 h (regimen A), 1,070 ng/ml at 4.4 h (regimen B), and 1,980 ng/ml at 6.0 h (regimen C). The steady state was achieved by day 13. Respective elimination half-lives on days 1 and 15 were 15 and 34 h (regimen A), 20.7, and 36.5 h (regimen B), and 25 and 41.7 h (regimen C), respectively. The areas under the plasma concentration versus time curves (0 to infinity) on day 1 were 1,320 (regimen A), 4,160 (regimen B), and 12,600 ng.h/ml (regimen C). With the exception of one patient on day 15 of regimen C, itraconazole was not detected in the urine. All data support dose-dependent pharmacokinetic behavior for itraconazole.


Inhibition of G-Protein βγ Signaling Decreases Levels of Messenger RNAs Encoding Proinflammatory Cytokines in T Cell Receptor-Stimulated CD4(+) T Helper Cells.
Inhibition of G-protein βγ (Gβγ) signaling was found previously to enhance T cell receptor (TCR)-stimulated increases in interleukin 2 (IL-2) mRNA in CD4(+) T helper cells, suggesting that Gβγ might be a useful drug target for treating autoimmune diseases, as low dose IL-2 therapy can suppress autoimmune responses. Because IL-2 may counteract autoimmunity in part by shifting CD4(+) T helper cells away from the Type 1 T helper cell (TH1) and TH17 subtypes towards the TH2 subtype, the purpose of this study was to determine if blocking Gβγ signaling affected the balance of TH1, TH17, and TH2 cytokine mRNAs produced by CD4(+) T helper cells. Gallein, a small molecule inhibitor of Gβγ, and siRNA-mediated silencing of the G-protein β1 subunit (Gβ1) were used to test the effect of blocking Gβγ on mRNA levels of cytokines in primary human TCR-stimulated CD4(+) T helper cells. Gallein and Gβ1 siRNA decreased interferon-γ (IFN-γ) and IL-17A mRNA levels in TCR-stimulated CD4(+) T cells grown under TH1-promoting conditions. Inhibiting Gβγ also decreased mRNA levels of STAT4, which plays a positive role in TH1 differentiation and IL-17A production. Moreover, mRNA levels of the STAT4-regulated TH1-associated proteins, IL-18 receptor β chain (IL-18Rβ), mitogen-activated protein kinase kinase kinase 8 (MAP3K8), lymphocyte activation gene 3 (LAG-3), natural killer cell group 7 sequence (NKG7), and oncostatin M (OSM) were also decreased upon Gβγ inhibition. Gallein also increased IL-4, IL-5, IL-9, and IL-13 mRNA levels in TCR-stimulated memory CD4(+) T cells grown in TH2-promoting conditions. Inhibiting Gβγ to produce these shifts in cytokine mRNA production might be beneficial for patients with autoimmune diseases such as rheumatoid arthritis (RA), Crohn's disease (CD), psoriasis, multiple sclerosis (MS), and Hashimoto's thyroiditis (HT), in which both IFN-γ and IL-17A are elevated.


The plasticity of the 7TMR signaling machinery and the search for pharmacological selectivity.
Components of 7TMR signaling machinery once considered as rigid, fixed and inflexible entities, operating in a one-dimensional way, homogeneous spatially and temporally, are now proved to be structurally plastic, flexible and dynamic in space and time. 7TMRs are thought to exist as ensembles of multiple, inter-convertible, pre-existing conformations and this conformational diversity provides a structural plasticity and the molecular mechanism for the functional diversity of 7TMRs. Furthermore, 7TMRs appear to function not as monomers, but rather as higher order structures, within which allosteric mechanisms affect ligand binding, G protein selection and receptor mobility and signalling of GPCR protomers. Moreover, their function is regulated through compartmentalization of G protein receptor and effector molecules into specialized membrane microdomains, such as lipid rafts and caveolae. Different permutations of receptor localization and translocation in membrane microdomains and internalization patterns, employed by 7TMRs regulate their signalling. Finally, the previous clear distinction between cell signalling and endocytic membrane trafficking has been blurred by evidence that these processes are intertwined and bidirectionally linked. 7TMRs normally signaling from the plasma membrane can elicit signaling cascades from an intracellular location, with distinct biochemical outputs. All these developments have moved the search for selective drugs beyond the mere design of receptor subtype-selective ligands targeting their orthosteric site. Even the notion of ligand has been expanded, so as to include "superagonists" ", acting as "super" 7TMR agonists to confer sustained endosomal signaling and biased agonists targeting GPCRs signalling from intracellular sites, as well as pharmacochaperones restoring insufficiently folded or misfolded receptors. The plasticity of the 7TMR signaling machinery and its dynamics in space and time will most probably impact further on the search for 7TMR drug selectivity.


Changes in cooperation and self-other differentiation during the second year.
Cooperation in peer interaction emerges during the second half of the second year. A consideration of the skills and knowledge entailed in these early forms of cooperation suggests that young children's emerging ability to differentiate self from other as causal agents may relate to their ability to coordinate behavior with age mates toward a common goal. Children at 12, 18, 24, and 30 months were observed in same-age, same-sex dyads (8 dyads per age) while attempting to solve a simple cooperation problem. They were also individually administered an elicited imitation task used to index decentration, or self-other differentiation. No 12-month-old dyad could cooperate, 18-month-olds did so infrequently and apparently accidentally, whereas 24- and 30-month-olds were able to coordinate behavior with one another quickly and effectively. Children who were better able to accommodate their behavior to one another during cooperation also represented the agency of others at a more advanced, decentered level.


Understanding counterfactuals in transparent and nontransparent context: An event-related potential investigation.
Counterfactuals describe imagined alternatives to reality that people know to be false. Successful counterfactual comprehension therefore requires people to keep in mind both an imagined hypothetical world and the presupposed real world. Counterfactual transparency, that is, the degree to which a context makes it easy to determine counterfactuality, might affect semantic processing. This might especially be the case for languages like Chinese which lack dedicated counterfactual markers and therefore are more context-dependent. Using event-related potentials, this study investigates the role of counterfactual transparency on the comprehension of Chinese counterfactuals. For transparent contexts (e.g., "If everything in the world could go back in time . . ."), in which the information needed to identify counterfactuality is highly accessible, discourse incongruent words elicited P600 effects. In contrast, for nontransparent contexts (e.g., "If better preparations were made at that time . . .") in which readers must attend to specific discourse context and engage pragmatic information to arrive at the counterfactual interpretation, discourse incongruencies gave rise to N400 effects. These findings suggest that (a) provided a constraining context, semantic processing is not disrupted by the dual nature of counterfactuality (i.e., readers can rapidly make contextually appropriate inferences to interpret subsequent narratives) and (b) the degree of transparency of the counterfactual can affect the nature of subsequent semantic processing. Our findings support the usage-based view that Chinese counterfactual comprehension is highly context-dependent and pragmatics-driven. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


[Epidemiologic study of the risk of tuberculosis recurrence in the Bas-Rhin, 1967-77].
The authors report an "analytic epidemiological" study, showing the risk of relapse in tuberculous patients in the Bas-Rhin in 1967 to 1977. This was possible thanks to statutory notification of tuberculous cases, with an index updated by annual returns. The study relates to French subjects notified as new cases of all forms of active tuberculosis between 1967 to 1975, going out the active file on or before 31.12.1976, and followed until 31.12.1977 after the cessation of chemotherapy. Amongst those gone out the active list, 5% were lost to follow up, 15% died, and 80% were alive (and bacteriologically inactive) at the end of treatment. In this latter group cases of relapse were observed between 1969 and 1977. Thus, 3,327 patients were at risk a relapse during the first year, and when the study was completed after 8 years, 76 relapses were observed. Two methods were used to calculate the risk of relapse. The first reported the number of relapses (76) to the number of people-years (14,500 for the 3,327 patients). The mean annual risk was 5,24%. The role of certain risk factors for a relapse could be identified more precisely: Sex, 5,43% in men and 3,19% in women, Age, 4,76% before 55, 8.36% after, Initial bacteriology 5,9% for positive cases and 3,27% for negative cases. Use of Rifampicin, 3,9% for users, 5,8% for non-users. The second calculation was based on actuarial method, giving the annual and cumulative risk as a function of the exposed population each year, for the period of the study.(ABSTRACT TRUNCATED AT 250 WORDS)


Development of a course based on BEAM robots to enhance STEM learning in electrical, electronic, and mechanical domains.
In this work, BEAM robotics is proposed to enhance the STEM knowledge and skills of engineering students in the electrical, electronic, and mechanical domains. To evaluate the proposal, a course is designed and implemented based on a curriculum with objectives and learning activities centered on the design, construction, and operation of the BEAM robots. In addition, the connection between this proposal and computational thinking is explored. Students learn to recognize each part of robots and how they are related, abstract useful information from an electronic scheme and concretize it in a machine by systematizing their behavior. In addition, thanks to an evaluation of the behavior of the robot, identify the faults and apply the solution, as in the debugging process carried out in software programming. It should be added that BEAM robotics has a sustainable and low-cost aspect, which is used in learning activities where Waste Electrical and Electronic Equipment (WEEE) is recycled, and students are taught to value and integrate these parts into the design of the robots. A pre and post survey and a respective statistical analysis and evaluation of curricular activities are presented as evidence of the improvement observed in students' STEM knowledge and skills. In general, the results show that this new teaching tool can promote the STEM curriculum in engineering students and motivate implementation, as a new educational robot, at other academic levels such as secondary and pre-secondary education.


Polymerase chain reaction/denaturing gradient gel electrophoresis (PCR/DGGE): sensitivity, band pattern analysis, and methodologic optimization.
Polymerase chain reaction/denaturing gradient gel electrophoresis (PCR/DGGE) has been gaining popularity as a preferred method to determine the clonality of T-cell populations in small or sparsely infiltrated specimens such as skin biopsies. T-cell receptor (TCR)-gamma gene rearrangements are amplified using nested consensus primers in two rounds of PCR and then are separated by DGGE. Sensitivity is better than with conventional Southern blot analysis but not fully defined. In addition to a discrete primary band resulting from a monoclonal TCR-gamma gene rearrangement, there are often weaker secondary bands of unknown origin. Our goals were to define the PCR/DGGE clonal detection threshold, determine the genesis of the multiband pattern, and optimize methods to minimize extraneous bands. Titration studies showed that the sensitivity of PCR/DGGE for detecting clonal T-cell DNA is affected by the polyclonal T-cell content of the background DNA. The detection threshold is 0.001% using keratinocyte DNA as diluent but only 1% with tonsil DNA. Analysis of monoclonal T-cell lines showed that multiple bands can be produced by a single TCR-gamma gene rearrangement. Mixing of inner and outer primer pair PCR products showed that this is an artifact resulting from different sized PCR products produced during the two rounds of nested PCR required for optimal specificity. Repeat DGGE of isolated bands ruled out variable mobility of partially melted PCR products. Reduction of first round PCR product used as second round target from 10 microl to 1 microl, or a decrease of first round primers from 40 pmole to 5 pmole, resulted in diminished secondary bands without compromising primary band intensity. These results show that: 1) PCR/DGGE has a realistic clonal detection threshold of 0.1% to 1%, 2) multiple bands are consistent with a monoclonal T-cell population, and 3) conditions can be optimized to minimize artifactual secondary bands.


Efficacy and Safety of Antiplatelet Agents for Adult Patients With Ischemic Moyamoya Disease.
Background: The use of antiplatelet agents in ischemic moyamoya disease (MMD) is controversial. This study aimed to investigate the effectiveness and safety of antiplatelet therapy compared with conservative treatment and surgical revascularization in ischemic MMD patients. Methods: Ischemic MMD patients were retrospectively enrolled from eight clinical sites from January 2013 to December 2018. Follow-up was performed through clinical visits and/or telephone interviews from first discharge to December 2019. The primary outcome was the episodes of further ischemic attacks, and the secondary outcome was the individual functional status. Risk factors for future stroke were identified by the LASSO-Cox regression model. Propensity score matching was applied to assemble a cohort of patients with similar baseline characteristics using the TriMatch package. Results: Among 217 eligible patients, 159 patients were included in the analyses after a 1:1:1 propensity score matching. At a mean follow-up of 33 months, 12 patients (7.5%) developed further incident cerebral ischemic events (surgical:antiplatelet:conservative = 1:3:8; p = 0.030), 26 patients (16.4%) developed a poor functional status (surgical:antiplatelet:conservative = 7:12:7; p = 0.317), and 3 patients (1.8%) died of cerebral hemorrhage (surgical:antiplatelet:conservative = 1:2:0; p = 0.361). The survival curve showed that the risk of further cerebral ischemic attacks was lowest with surgical revascularization, while antiplatelet therapy was statistically significant for preventing recurrent risks compared with conservative treatment (χ2 = 8.987; p = 0.011). No significant difference was found in the functional status and bleeding events. The LASSO-Cox regression model revealed that a family history of MMD (HR = 6.93; 95% CI: 1.28-37.52; p = 0.025), a past history of stroke or transient ischemic attack (HR = 4.35; 95% CI: 1.09-17.33; p = 0.037), and treatment (HR = 0.05; 95% CI: 0.01-0.32; p = 0.001) were significantly related to the risk of recurrent strokes. Conclusions: Antiplatelet agents were effective and safe in preventing further cerebral ischemic attacks in adult patients with ischemic MMD. They may be a replacement therapy for patients with surgical contraindications and for patients prior to revascularization.


LIGHT AND ELECTRON MICROSCOPE OBSERVATIONS ON THE TUBE-DWELLING DIATOM AMPHIPLEURA RUTILANS (TRENTEPOHL) CLEVE.
The tubular sheath of Amphipleura rutilans is composed of small fibrils 20-25 A in diameter. The endotubular environment is further partitioned by individual cell sheath formation. The tube boundary seems to be an effective barrier against damage from extreme salinity changes, but does not prevent passage of EDTA, neutral red, surfactants, or caffeine. Terminal swellings develop on growing tips during extremely cold periods. These give rise to 2 or more separate growing tips which proceed to form regular tubes. There was no evidence for sexual reproduction or auxospore formation within the terminal swellings, even though adjacent swellings fused. Bacterial, algal, and fungal epiphytes were abundant on the extratubular surface. Hollow lipid bodies occurred in many cells. A rimring strand of presumed DNA occurs in the chloroplast. It is proposed that the tube material serves not only as a protective boundary but also functions as an active nutrient trap.


Patient Portal Preferences: Perspectives on Imaging Information.
Patient portals have the potential to provide content that is specifically tailored to a patient's information needs based on diagnoses and other factors. In this work, we conducted a survey of 41 lung cancer patients at an outpatient lung cancer clinic at the medical center of the University of California Los Angeles, to gain insight into these perceived information needs and opinions on the design of a portal to fulfill them. We found that patients requested access to information related to diagnosis and imaging, with more than half of the patients reporting that they did not anticipate an increase in anxiety due to access to medical record information via a portal. We also found that patient educational background did not lead to a significant difference in desires for explanations of reports and definitions of terms.


Small cardioactive peptide gene: structure, expression and mass spectrometric analysis reveals a complex pattern of co-transmitters in a snail feeding neuron.
The small cardioactive peptides (SCPs) are an important group of neural cotransmitters in molluscs where they are known to play both central and peripheral modulatory roles in the control of feeding behaviour. Here we show that in the snail Lymnaea the SCP gene exists in one interrupted copy that produces a single species of transcript which encodes a prepropeptide containing two structurally related SCPs SGYLAFPRMamide (SCP(A)) and pQNYLAFPRMamide (SCP(B)). In situ hybridization was used to localize expression specifically to the soma of several types of motoneurons in the feeding system of Lymnaea, including the giant B2 foregut motoneurons. The peptide content of individual B2 cell bodies was analysed by matrix-assisted laser desorption/ionization mass spectrometry and the structures of the SCPs predicted from the cloned gene were confirmed in these cells by post-source decay fragmentation analysis. Identical stimulatory activity for the two SCP peptides was demonstrated by their application to the isolated foregut, suggesting that their co-release from the B2 cells may play an important part in the co-modulation of gut motility, together with acetylcholine and the myomodulin family of peptides.


Cytokeratin labeling of breast cancer cells extracted from paraffin-embedded tissue for bivariate flow cytometric analysis.
Flow cytometric analyses of DNA content and proliferative fractions have been found to be important prognostic indicators in a variety of human tumors. However, variability in reported results and interlaboratory differences in single-parameter DNA measurements have impeded the broader use of this methodology. Multiparameter DNA analysis, especially that which allows the ploidy and cell cycle measurements to be targeted specifically to tumor cells, may improve the quality and reliability of these measurements. Cytokeratin labeling simultaneously with DNA allows the identification of the malignant epithelial cells within tumor samples that have been microdissected for tumor enrichment and, thus, can increase the accuracy of tumor ploidy and cell cycle measurements. There are a limited number of reports of cytokeratin labeling of paraffin-extracted cells, and results with standard preparation procedures can be highly variable. We have developed an improved technique for cytokeratin labeling of paraffin-extracted cells that is based on predigestion of tissue with collagenase prior to brief pepsin digestion. This two-step enzymatic digestion produces a superior cytokeratin vs. DNA bivariate analysis, with increased intensity and greater uniformity of cytokeratin labeling. This method should increase the accuracy of ploidy and proliferation measurements from paraffin-embedded tissue both for retrospective studies and in clinical settings in which only fixed tissue is available.


Evaluation of Dual Tobacco Smoking (Water Pipe and Cigarettes) and Associated Factors in Adults in Tehran.
Concurrent use of tobacco products such as cigarettes and water pipes may be associated with increased risk of nicotine dependence and smoking-related complications. Accurate statistics are not available regarding the prevalence of water pipe use or concurrent use of cigarettes and water pipe in the Iranian population. Thus, this study sought to assess the prevalence of concurrent use of cigarettes and water pipes and their related factors in Iran. This cross-sectional study was conducted on Tehran residents over 15 years of age, who were selected via cluster, multi-stage randomized sampling, from different geographical districts of Tehran between November and December 2014. The data were collected using the water pipe section of the Global Adult Tobacco Survey (GATS) questionnaire. A total of 1,830 individuals participated in this study, 243 (13.3%) of which exclusively used water pipes, 76 (4.2%) used both cigarettes and water pipes, and 120 (6.6%) exclusive smoked cigarettes. Of those who used both cigarettes and water pipes, 86.8% were men and 13.2% were women (P < 0.001). The mean age of those who only used water pipes was 28.01 ± 8.7 years while the mean age of those who used both water pipes and cigarettes was 33.1 ± 1.1 (P < 0.001). Male sex (adj. OR: 3.8) and older age (adj. OR: 1.06) increased the odds of using both tobacco products. The prevalence of concurrent use of cigarettes and water pipes and that of exclusive water pipe use were 4.2% and 13.3%, respectively. The prevalence of exclusive cigarette smoking was 6.1%. Those who smoked both cigarettes and water pipes had a higher mean age than those who exclusively used water pipes and they were mostly men. Among those who used cigarettes and water pipes, the mean age at which they began using water pipes was lower than the mean age at which they began smoking cigarettes. In other words, dual smokers started water pipe smoking sooner than cigarette smoking. Future studies with different methodologies are required to further scrutinize the relationship between water pipe and cigarette smoking. Smoking cessation programs must specifically target dual smokers.


Metastatic melanoma concurrent to the urinary bladder and endometrium: Case report.
We report a case of a 47-year-old woman with a history of acral lentiginous melanoma, left great toe subungual primary, metastatic to the lungs, brain, and spleen, presenting with lower urinary tract symptoms. She had also palpated and removed a dark mass from her vaginal canal. Magnetic resonance imaging (MRI) showed polypoidal lesions within the urinary bladder and the endometrium. The suspected differential diagnosis was endometrial and bladder melanoma metastases, based on her cancer history and MRI findings. The patient underwent cystoscopy with transurethral resection of the bladder tumor and endometrial biopsy, and pathology was consistent with metastatic melanoma. Bladder and endometrial metastatic lesions are exceedingly rare. Herein, we describe an unusual case of metastatic melanoma concurrent to the urinary bladder and endometrium.


A new paradigm shift in antithrombotic therapy.
Decades after the introduction of oral anti-coagulants namely the vitamin K antagonist (VKA) Warfarin and antiplatelet agents such as Aspirin and Plavix, new classes of direct, small molecule, novel oral anti-coagulant medications and antiplatelet P2Y12 receptor inhibitors have recently become available. For the novel oral anticoagulants (NOAC), these agents can be separated by direct thrombin inhibitors such as Dabigatran and direct Factor Xa inhibitors such as Rivaroxaban and Apixaban. For next generation antiplatelet agents such as Ticagrelor and Prasugrel, these new P2Y12 receptor inhibitors form the cornerstone of therapy for patients with acute coronary syndrome (ACS) or undergoing percutaneous interventions. These novel oral antithrombotics are revolutionizing the field of stroke prevention, atrial fibrillation (AF), the management of venous thromboembolism (VTE) and treatment of ACS. This article reviews the current research developed in order to identify therapeutic effects and establish net clinical benefits of these new oral antithrombotics.


High-performance liquid chromatographic determination of total thiamine in human plasma for oral bioavailability studies.
A high-performance liquid chromatographic method for the determination of total thiamine in plasma was developed for a relative bioavailability comparison of two oral thiamine preparations. After separation of thiamine mono- and diphosphates with the phosphatase enzyme, the total thiamine was subjected to reversed-phase high-performance liquid chromatography, postcolumn oxidized to thiochrome with K3[Fe(CN)6], and detected by fluorescence. In the bioavailability study, 16 human subjects were put on a low-thiamine diet for 3 days. On the 2nd and 3rd days, 14 blood samples per subject and day were taken at the same times each day. Drug administration did not take place until the 3rd day. After deduction of the native concentrations of total thiamine detected on the 2nd day (mean value of approximately 7 ng/mL), the post-treatment pharmacokinetic parameters were determined (two different preparations, each with 200 mg of thiamine.HCl).


Molecular cloning of a human glycophorin B cDNA: nucleotide sequence and genomic relationship to glycophorin A.
Here we describe the isolation and nucleotide sequence of a human glycophorin B cDNA. The cDNA was identified by differential hybridization of synthetic oligonucleotide probes to a human erythroleukemic cell line (K562) cDNA library constructed in phage vector lambda gt10. The nucleotide sequence of the glycophorin B cDNA was compared with that of a previously cloned glycophorin A cDNA. The nucleotide sequences encoding the NH2-terminal leader peptide and first 26 amino acids of the two proteins are nearly identical. This homologous region is followed by areas specific to either glycophorin A or B and a number of small regions of homology, which in turn are followed by a very homologous region encoding the presumed membrane-spanning portion of the proteins. We used RNA blot hybridization with both cDNA and synthetic oligonucleotide probes to prove our previous hypothesis that glycophorin B is encoded by a single 0.5- to 0.6-kb mRNA and to show that glycophorins A and B are negatively and coordinately regulated by a tumor-promoting phorbol ester, phorbol 12-myristate 13-acetate. We established the intron/exon structure of the glycophorin A and B genes by oligonucleotide mapping; the results suggest a complex evolution of the glycophorin genes.


Clinical Outcomes of Severe COVID-19 Patients Admitted to an Intermediate Respiratory Care Unit.
Introduction: Many severe COVID-19 patients require respiratory support and monitoring. An intermediate respiratory care unit (IMCU) may be a valuable element for optimizing patient care and limited health-care resources management. We aim to assess the clinical outcomes of severe COVID-19 patients admitted to an IMCU. Methods: Observational, retrospective study including patients admitted to the IMCU due to COVID-19 pneumonia during the months of March and April 2020. Patients were stratified based on their requirement of transfer to the intensive care unit (ICU) and on survival status at the end of follow-up. A multivariable Cox proportional hazards method was used to assess risk factors associated with mortality. Results: A total of 253 patients were included. Of them, 68% were male and median age was 65 years (IQR 18 years). Ninety-two patients (36.4%) required ICU transfer. Patients transferred to the ICU had a higher mortality rate (44.6 vs. 24.2%; p < 0.001). Multivariable proportional hazards model showed that age ≥65 years (HR 4.14; 95%CI 2.31-7.42; p < 0.001); chronic respiratory conditions (HR 2.34; 95%CI 1.38-3.99; p = 0.002) and chronic kidney disease (HR 2.96; 95%CI 1.61-5.43; p < 0.001) were independently associated with mortality. High-dose systemic corticosteroids followed by progressive dose tapering showed a lower risk of death (HR 0.15; 95%CI 0.06-0.40; p < 0.001). Conclusions: IMCU may be a useful tool for the multidisciplinary management of severe COVID-19 patients requiring respiratory support and non-invasive monitoring, therefore reducing ICU burden. Older age and chronic respiratory or renal conditions are associated with worse clinical outcomes, while treatment with systemic corticosteroids may have a protective effect on mortality.


Parsonage-Turner syndrome and cytomegalovirus disease.
Neuralgic amyotrophy (NA), also known as Parsonage-Turner syndrome (PTS), is a distinct idiopathic immune-mediated neuritis of the brachial plexus, characterized by sudden attacks of severe neuropathic pain usually in the shoulder and/or arm, followed by progressive neurologic deficits, including weakness, atrophy, and occasionally sensory abnormalities. Pathogenesis is assumed to be multifactorial, and several observations support the hypothesis of an immune-triggering event preceding PTS, most frequently infections. A literature review reveals a variety of clinical presentations and courses. Various microorganisms preceding PTS have been documented. The authors report a case of PTS related to cytomegalovirus infection with a review of the relevant literature. Special emphasis is placed on the most important infectious agents considered in the etiological list of PTS.


Agastinol and agastenol, novel lignans from Agastache rugosa and their evaluation in an apoptosis inhibition assay.
Investigation of the whole plant of Agastache rugosa resulted in the isolation of two new lignan compounds. Their structures were elucidated as (8S,7'R,8'S)-4-hydroxybenzoic acid 4-(4-hydroxy-3-methoxybenzyl)-2-(4-hydroxy-3-methoxyphenyl)tetrahydrofuran-3-ylmethyl ester (agastinol, 1) and (7'R,8'S)-4-hydroxybenzoic acid 4-(hydroxy-3-methoxybenzylidene)-2-(4-hydroxy-3-methoxyphenyl)-tetrahydrofuran-3-ylmethyl ester (agastenol, 2). Agastinol and agastenol inhibited etoposide-induced apoptosis in U937 cells with IC50 values of 15.2 and 11.4 microg/mL, respectively.


Validation study of the Food Craving Questionnaire-Trait: Reduced in Greek population.
Food craving (FC) is a behavior which results in an increased total energy intake. It is considered to be a multivariate outcome of (neuro)biologicals and environmental alterations that mostly impacts in bulimic and binge-eating behaviors. FC measurement questionnaires vary in the literature, with the most widely used being the Food Craving Questionnaire-State, Trait and Traitreduced. The purpose of the current study was the validation of the Food Craving Questionnaire-Traitreduced (FCQ-T-r) in Greek population. Along with the FCQ-T-r, the Eating Attitude Test-26 (EAT-26) was used for the evaluation of nutritional behavior, as well as the General Health Questionnaire-28 (GHQ-28) for assessing the health of the sample and a self-generated questionnaire for demographic information and anthropometric measurements. The Principal Component Analysis (PCA) confirmation was made with Monte-Carlo Parallel Analysis (PA). The FCQ-T-r showed a high internal consistency (Cronbach's Alpha=0.927) explaining up to 62.5% variance of the sample. The PCA indicated a two-factor solution (Thoughts & Emotions and Lack of Control) highly correlated with 14 questions in contrast from the original version which consists one-factor with 15 questions. The Monte-Carlo PA confirmed these findings. Additionally, statistical significantly and positively weak to moderate correlations have been observed between the total score of FCQ-T-r with the EAT-26 (r=0.28), EAT-Dieting (r=0.27), EAT-Bulimia (r=0.41) and the GHQ (total score and sub-scales r=0.17 to 0.24). Concurrent, its sub-scales Thoughts & Emotions and Lack of Control were statistical significantly stronger correlated with EAT-Dieting (r=0.24/0.38), EAT-Bulimia (r=0.27/0.37) and lower with GHQ and its sub-scales (total score and sub-scales r=0.15 to 0.29). The above results suggest fair psychometric properties and validity of the FCQ-T-r that could be a useful tool for indicating-measuring the tendency of food craving and possibly the eating behavior.


Resting-state functional network correlates of psychotic symptoms in schizophrenia.
Schizophrenia has been associated with aberrant intrinsic functional organization of the brain but the relationship of such deficits to psychopathology is unclear. In this study, we investigated associations between resting-state networks and individual psychopathology in sixteen patients with paranoid schizophrenia and sixteen matched healthy control participants. We estimated whole-brain functional connectivity of multiple networks using a combination of spatial independent component analysis and multiple regression analysis. Five networks (default-mode, left and right fronto-parietal, left fronto-temporal and auditory networks) were selected for analysis based on their involvement in neuropsychological models of psychosis. Between-group comparisons and correlations to psychopathology ratings were performed on both spatial (connectivity distributions) and temporal features (power-spectral densities of temporal frequencies below 0.06 Hz). Schizophrenia patients showed aberrant functional connectivity in the default-mode network, which correlated with severity of hallucinations and delusions, and decreased hemispheric separation of fronto-parietal activity, which correlated with disorganization symptoms. Furthermore, the severity of positive symptoms correlated with functional connectivity of fronto-temporal and auditory networks. Finally, default-mode and auditory networks showed increased spectral power of low frequency oscillations, which correlated with positive symptom severity. These results are in line with findings from studies that investigated the neural correlates of positive symptoms and suggest that psychopathology is associated with aberrant intrinsic organization of functional brain networks in schizophrenia.


A study on the outcome of percutaneous transluminal renal angioplasty in patients with renal failure.
The indications for percutaneous transluminal renal angioplasty (PTRA) in renovascular disease, as well as its benefits, remain a matter of debate. The aim of this study was to evaluate the outcome of angioplasty and to identify risk factors associated with less successful outcomes in patients with atheromatous renal artery stenosis and renal failure of varying degrees. The results of PTRA were analyzed retrospectively in 144 patients with serum creatinine levels of >130 micromol/l. Patients were divided into 5 groups according to their indication for angioplasty: (1) deteriorating renal function; (2) accelerating hypertension; (3) a combination of 1 and 2; (4) peripheral vascular disease, and (5) miscellaneous conditions. The baseline mean (+/- SD) systolic and diastolic blood pressures of the entire group were lowered from 180 +/- 32 and 95 +/- 16 mm Hg to 162 +/- 23 and 86 +/- 12 mm Hg, respectively (p < 0.0005), 12 months after angioplasty. The blood pressure level was unaffected by angioplasty in patients with claudication. The mean number of antihypertensive drugs was reduced in the group with accelerating hypertension from 2.9 +/- 0.8 to 2.4 +/- 1.2 (p = 0.019), and in the group with unilateral renal artery stenosis and two kidneys from 2.4 +/- 1.0 to 1.8 +/- 1.1 (p = 0.002), 12 months after PTRA. Glomerular filtration rate at 3-month follow-up had increased from 23 +/- 11 to 27 +/- 14 ml/min/1.73 m(2) (p = 0.021) in group 1, from 25 +/- 11 to 28 +/- 14 ml/min/1.73 m(2) (p = 0.031) in the combined group of patients consisting of groups 1 and 3, and from 32 +/- 13 to 35 +/- 14 ml/min/1.73 m(2) (p = 0.019) in the group with unilateral renal artery stenosis. No statistically significant difference was found in any of these 3 groups 1 year after angioplasty. The first patient group had an increased prevalence of cardiovascular disease, aortic aneurysm, carotid occlusive disease, and peripheral vascular disease compared to the other patient groups (p < 0.05). Patients with baseline creatinine levels of >300 micromol/l had a lower survival rate at 12, 60, and 120 months after PTRA than patients with serum creatinine levels of <300 micromol/l (p < 0.005). Survival was also lower in patients with bilateral renal artery stenosis and those with a single kidney, compared to patients with a unilateral stenosis at both 5 and 10 years after PTRA (p < 0.05). Regression analysis of predictor variables of mortality rate showed that the relative risk (RR) associated with increased serum creatinine was 4.7 (CI 2.0-11.0; p < 0.0005). The RR for older patients was 1.1 (CI 1.0-1.2; p = 0.008), and the RR for former smokers was 6.0 (CI 1.6-24.0; p = 0.009). The results of the present study indicate that glomerular filtration can be improved in patients who primarily undergo angioplasty to rescue renal function. Renal function with creatinine levels of >300 micromol/l was associated with a lower survival rate. It is, therefore, possible that patients selected after a thorough evaluation of their renal function and comorbid disease factors may benefit from PTRA, even when the indication for angioplasty is to salvage renal function.


Residual Disease after Operative Hysteroscopy in Patients with Endometrioid Endometrial Cancer Associated with Polyps.
To evaluate the presence of residual disease in the uterine specimen after hysteroscopic polypectomy or polyp biopsy in patients with endometrioid endometrial cancer (EC). We analyzed a series of 104 patients (92 cases from the Hospital AC Camargo and 12 from the Hospital do Servidor Público Estadual de São Paulo) with polyps that were diagnosed by hysteroscopy, showing endometrioid EC associated with the polyp or in the final pathological specimen. Patients underwent a surgical approach for endometrial cancer from January 2002 to January 2017. Their clinical and pathological data were retrospectively retrieved from the medical records. In 78 cases (75%), the polyp had EC, and in 40 (38.5%), it was restricted to the polyp, without endometrial involvement. The pathologic stage was IA in 96 cases (92.3%) and 90 (86.5%) had histologic grade 1 or 2. In 18 cases (17.3%), there was no residual disease in the final uterine specimen, but only in 9 of them the hysteroscopy suggested that the tumor was restricted to the polyp. In 5 cases (4.8%) from the group without disease outside of the polyp during hysteroscopy, myometrial invasion was noted in the final uterine specimen. This finding suggests the possibility of disease extrapolation through the base of the polyp. Patients with endometrioid EC associated with polyps may have the tumor completely removed during hysteroscopy, but the variables shown in the present study could not safely predict which patient would have no residual disease.


Increased peroxisome proliferator-activated receptor γ expression levels in visceral adipose tissue, and serum CCL2 and interleukin-6 levels during visceral adipose tissue accumulation.
This study was conducted to determine the mRNA and protein expression levels of peroxisome proliferator-activated receptors (PPARs) in visceral adipose tissue, as well as serum adipokine levels, in Sprague Dawley rats. The rats were fed either a normal (control rats) or excessive (experimental rats) intake of food for 8 or 16 weeks, then sacrificed, at which time visceral and subcutaneous adipose tissues, as well as blood samples, were collected. The mRNA and protein expression levels of PPARs in the visceral adipose tissues were determined using reverse transcription-polymerase chain reaction and Western blotting, respectively. In addition, the levels of adipokines in the serum samples were determined using commercial ELISA kits. The results revealed that at 8 weeks, the mass of subcutaneous adipose tissue was higher than that of the visceral adipose tissue in the experimental rats, but the reverse occurred at 16 weeks. Furthermore, at 16 weeks the experimental rats exhibited an upregulation of PPARγ mRNA and protein expression levels in the visceral adipose tissues, and significant increases in the serum levels of CCL2 and interleukin (IL)-6 were observed, compared with those measured at 8 weeks. In conclusion, this study demonstrated that the PPARγ expression level was likely correlated with serum levels of CCL2 and IL-6, molecules that may facilitate visceral adipose tissue accumulation. In addition, the levels of the two adipokines in the serum may be useful as surrogate biomarkers for the expression levels of PPARγ in accumulated visceral adipose tissues.


Misconduct versus honest error and scientific disagreement.
Researchers sometimes mistakenly accuse their peers of misconduct. It is important to distinguish between misconduct and honest error or a difference of scientific opinion to prevent unnecessary and time-consuming misconduct proceedings, protect scientists from harm, and avoid deterring researchers from using novel methods or proposing controversial hypotheses. While it is obvious to many researchers that misconduct is different from a scientific disagreement or simply an inadvertent mistake in methods, analysis or misinterpretation of data, applying this distinction to real cases is sometimes not easy. Because the line between misconduct and honest error or a scientific dispute is often unclear, research organizations and institutions should distinguish between misconduct and honest error and scientific disagreement in their policies and practices. These distinctions should also be explained during educational sessions on the responsible conduct of research and in the mentoring process. When researchers wrongfully accuse their peers of misconduct, it is important to help them understand the distinction between misconduct and honest error and differences of scientific judgment or opinion, pinpoint the source of disagreement, and identify the relevant scientific norms. They can be encouraged to settle the dispute through collegial discussion and dialogue, rather than a misconduct allegation.


Platelet derived malonyldialdehyde production in patients with thalassaemia major.
Malonyldialdehyde, a product of membrane lipid peroxidation, was measured in the platelets of 16 normal subjects after stimulation with a variety of aggregating and stimulating agents. Nethylmaleimide and hydrogen peroxide generated the largest amounts of malonyldialdehyde. These agents were used to stimulate platelets from 11 patients with thalassaemia major suffering from iron overload due to repeated transfusion. Mean malonyldialdehyde concentrations were the same in normal subjects as in thalassaemic patients, but high concentrations were recorded in patients with severe iron overload. There was a highly significant correlation between malonyldialdehyde and serum ferritin concentrations in all thalassaemic patients. Platelet derived malonyldialdehyde may be a useful test of continuing membrane damage in patients with iron overload.


The desensitization of the embryonic mouse muscle acetylcholine receptor depends on the cellular environment.
The rate of desensitization of nicotinic acetylcholine (ACh) receptor (nAChR), an important characteristic of nAChR function, was studied in myotubes of the mouse C2C12 cell line at different times after fusion, by measuring the decay of ACh-evoked currents (IACh) under various patch-clamp configurations. We observed a progressive slowing of IACh decay rate (half-decay time rose from about 0.5 s to over 5 s) in myotubes of increasing size (i.e. age) under all experimental conditions, except in outside-out patches, when IACh decayed as fast as in the smallest myotubes. Single-channel conductance (about 35 pS) and open time (about 3.5 ms), measured in outside-out and cell-attached patches, were independent of myotube size. In Xenopus oocytes injected with poly(A+)RNA extracted from C2C12 myoblasts or mature myotubes, IACh decay was about 50 times slower than in myotubes. Neither cAMP-dependent nor diacylglycerol-dependent protein kinases, actin nor microtubule polymerization state influenced IACh decay. Our data indicate that the cellular environment, but not readily dialysable cytosolic factors, markedly influences the functional behaviour of nAChR.


β-Glucan-Rich Extract of Gray Oyster Mushroom, Pleurotus pulmonarius, Improves Object Recognition Memory and Hippocampus Morphology in Mice Fed a High-Fat Diet.
Obesity may cause behavioral alterations, while maternal obesity can contribute to metabolic disorders in subsequent generations. The effect of β-glucan-rich Pleurotus pulmonarius (βgPp) was investigated on mouse neurobehavior and hippocampus and its offspring's hippocampus development. Female ICR mice were fed with normal diet (ND), ND with βgPp, high-fat diet (HFD), or HFD with βgPp for 3 months followed by behavioral test and mating. Immunohistochemistry for the expression of neuronal nuclear protein (NeuN) and ionized calcium binding adaptor molecule-1 (Iba-1) in the hippocampus was carried out. βgPp significantly enhanced short-term object recognition memory in HFD-fed mice. βgPp also ameliorated the histological alterations and neuronal loss and increased Iba-1-positive microglia in the hippocampus regions of HFD-fed mice and their male offspring. These findings demonstrated that βgPp supplementation attenuated the effects of HFD on object recognition memory and the alterations on the hippocampal regions of maternal mice and their male offspring.


Personal and contextual factors in the escalation of driving after drinking across the college years.
College students continue to drive after drinking at alarmingly high rates. Age trends suggest that driving after drinking increases from late adolescence across the college years, largely mirroring trends in binge drinking. Relatively little research, however, has examined change over time in driving after drinking among college students or tested whether some students might be at greater risk of escalations in driving after drinking. Using a sample of 1,833 nonabstaining students who completed surveys for seven semesters across the college years, we tested whether personal (i.e., age of drinking onset, gender, risk perceptions, and sensation seeking) and contextual (i.e., college residence) factors were associated with changes in driving after drinking. Using latent growth curve modeling, we found significant individual differences in rates of change in driving after drinking. Male students and students who began drinking earlier in life increased in driving after drinking more rapidly, whereas living in on-campus housing was associated with time-specific decreases in driving after drinking. These results demonstrate the value of considering driving after drinking from a longitudinal perspective and suggest possible avenues toward preventing the public health consequences of intoxicated driving.


CT angiography for living kidney donors: accuracy, cause of misinterpretation and prevalence of variation.
To determine the accuracy of the use of multi-detector row CT (MDCT) to predict vascular anatomy in living kidney donors and to reveal the prevalence of vascular variations in a Korean population. A total of 153 living kidney donors that had undergone preoperative CT and nephrectomy, either with open or laparoscopic surgery, were selected retrospectively. The initial CT results were compared with the surgical findings and repeated review sessions of CT scans were performed to determine the causes of mismatches in discordant cases. The accuracy of CT angiography was 95% to predict the number of renal vessels. Four arteries and two veins were missed during the initial CT interpretation due to perception errors (for two arteries and two veins) and technical limitations (two arteries). The prevalence of multiple renal arteries and veins, early branching of a renal artery and late confluence of a renal vein were 31%, 5%, 12%, 17%, respectively. The circumaortic renal vein and the bilateral inferior vena cava were found in two cases each (1.3%). One case (0.7%) each of a retroaortic renal vein and a supradiaphragmatic originated renal artery were found. MDCT provides a reliable method to evaluate the vascular anatomy and variations of living kidney donors.


Analysis of site-specific binding of (+/-)-anti-benzo[a]pyrene diol epoxide to restriction fragments of pBR322 DNA via photochemical mapping.
The binding sites and relative reactivity of (+/-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene [(+/-)-anti-BPDE] covalently bound to restriction fragments of pBR322 DNA are determined. (+/-)-anti-BPDE-modified DNA undergoes a photodissociation at the site of these adducts when irradiated with 355-nm laser light, resulting in a scission of the DNA sugar-phosphate backbone producing DNA fragments similar to those of Maxam-Gilbert sequencing reactions. The binding sites of (+/-)-anti-BPDE with each DNA base are determined by sequencing gel analysis of the BPDE-mediated photolysis and laser densitometry of the resulting banding patterns. This technique was used to analyze the binding of (+/-)-anti-BPDE to the 5' and 3' strands of the EcoRI/EcoRV and BamHI/SalI restriction fragments of pBR322 DNA. The reactivity of (+/-)-anti-BPDE to guanine bases within guanine-rich regions of DNA is enhanced by as much as a factor of 17 relative to the least reactive guanines which are flanked by non-guanine bases. The results also show enhanced photocleavage of the backbone corresponding to non-guanine bases in guanine-rich regions. These results suggest either that non-guanine basis in guanine-rich regions are more reactive than identical bases in other regions of the restriction fragment or that photocleavage of the backbone occurs adjacent to a BPDE-modified guanine. The binding profiles of (+/-)-anti-BPDE to pBR322 DNA at a binding density of 0.52 and 0.93 BPDE adduct per strand gave essentially identical binding patterns.(ABSTRACT TRUNCATED AT 250 WORDS)


Increasing rates of preterm twin births coincide with improving twin pair survival.
To examine trends in twin gestational age over time, with adjustment for potential confounding factors, and to assess twin pair mortality and respiratory support over time. Rates of preterm births, respiratory support, and neonatal mortality were calculated for 21,569 twin pairs born from 1980 to 2005 in Washington State, using birth certificate and hospital discharge data. Fetal death risks were determined on a "per-pair-at-risk" basis. While the proportion of twins born at 24-31 weeks remained stable at 8%, the proportion born at 32-36 weeks increased from 28% to 48%, and the proportion at 37-42 weeks declined from 64% to 44% (P<0.0001). Controlling individually for a variety of factors, such as maternal age, race, parity, and mode of delivery did not diminish the highly significant trend of increasing preterm births (P<0.0001 for each). Twin pair neonatal mortality decreased significantly through time (P<0.0001); however, the rate of pairs with one or both infants requiring oxygen or ventilation increased significantly through time (P<0.0001). Fetal death risks declined for term twins. The proportion of twins born at 32-36 weeks' gestation has increased over time, along with requirement for respiratory support. Twin pair mortality decreased from 1980 to 2005.


A map of open chromatin in human pancreatic islets.
Tissue-specific transcriptional regulation is central to human disease. To identify regulatory DNA active in human pancreatic islets, we profiled chromatin by formaldehyde-assisted isolation of regulatory elements coupled with high-throughput sequencing (FAIRE-seq). We identified approximately 80,000 open chromatin sites. Comparison of FAIRE-seq data from islets to that from five non-islet cell lines revealed approximately 3,300 physically linked clusters of islet-selective open chromatin sites, which typically encompassed single genes that have islet-specific expression. We mapped sequence variants to open chromatin sites and found that rs7903146, a TCF7L2 intronic variant strongly associated with type 2 diabetes, is located in islet-selective open chromatin. We found that human islet samples heterozygous for rs7903146 showed allelic imbalance in islet FAIRE signals and that the variant alters enhancer activity, indicating that genetic variation at this locus acts in cis with local chromatin and regulatory changes. These findings illuminate the tissue-specific organization of cis-regulatory elements and show that FAIRE-seq can guide the identification of regulatory variants underlying disease susceptibility.


SpiNNTools: The Execution Engine for the SpiNNaker Platform.
SpiNNaker is a massively parallel distributed architecture primarily focused on real time simulation of spiking neural networks. The largest realization of the architecture consists of one million general purpose processors, making it the largest neuromorphic computing platform in the world at the present time. Utilizing these processors efficiently requires expert knowledge of the architecture to generate executable code and to harness the potential of the unique inter-processor communications infra-structure that lies at the heart of the SpiNNaker architecture. This work introduces a software suite called SpiNNTools that can map a computational problem described as a graph into the required set of executables, application data and routing information necessary for simulation on this novel machine. The SpiNNaker architecture is highly scalable, giving rise to unique challenges in mapping the problem to the machines resources, loading the generated files to the machine and subsequently retrieving the results of simulation. In this paper we describe these challenges in detail and the solutions implemented.


Stabilization of the veratryl alcohol cation radical by lignin peroxidase.
Lignin peroxidase (LiP) catalyzes the H2O2-dependent oxidation of veratryl alcohol (VA) to veratryl aldehyde, with the enzyme-bound veratryl alcohol cation radical (VA.+) as an intermediate [Khindaria et al. (1995) Biochemistry 34, 16860-16869]. The decay constant we observed for the enzyme generated cation radical did not agree with the decay constant in the literature [Candeias and Harvey (1995) J. Biol. Chem. 270, 16745-16748] for the chemically generated radical. Moreover, we have found that the chemically generated VA.+ formed by oxidation of VA by Ce(IV) decayed rapidly with a first-order mechanism in air- or oxygen-saturated solutions, with a decay constant of 1.2 x 10(3) s-1, and with a second-order mechanism in argon-saturated solution. The first-order decay constant was pH- independent suggesting that the rate-limiting step in the decay was deprotonation. When VA.+ was generated by oxidation with LiP the decay also occurred with a first-order mechanism but was much slower, 1.85 s-1, and was the same in both oxygen- and argon-saturated reaction mixtures. However, when the enzymatic reaction mixture was acid-quenched the decay constant of VA.+ was close to the one obtained in the Ce(IV) oxidation system, 9.7 x 10(2) s-1. This strongly suggested that the LiP-bound VA.+ was stabilized and decayed more slowly than free VA.+. We propose that the stabilization of VA.+ may be due to the acidic microenvironment in the enzyme active site, which prevents deprotonation of the radical and subsequent reaction with oxygen. We have also obtained reversible redox potential of VA.+/VA couple using cyclic voltammetery. Due to the instability of VA.+ in aqueous solution the reversible redox potential was measured in acetone, and was 1.36 V vs normal hydrogen electrode. Our data allow us to propose that enzymatically generated VA.+ can act as a redox mediator but not as a diffusible oxidant for LiP-catalyzed lignin or pollutant degradation.


COVID-19: Implications for Supply Chain Management.
The COVID-19 pandemic has created global health and economic disruption. Hospitals and other healthcare providers have been hit particularly hard. While efforts to effectively treat and eradicate the coronavirus continue, so do the efforts of supply chains to support the provision of patient care in the event of a resurgence or future pandemic. Supply chain leaders must continuously evaluate their strategic and tactical positions to address critical supply needs. Whether the supply chain can meet expectations remains uncertain, given rolling supply shortages of personal protective equipment (PPE) and other medical-surgical supplies as healthcare providers resume prepandemic levels of operations. The ability to ensure a reliable, sustainable supply of critical PPE in the near term will remain a challenge. Longer-term substantive changes to the function and performance of healthcare supply chains will be necessary across multiple areas to meet demand more effectively during a crisis.


Complete obstruction of right proximal pulmonary artery: a case report.
A fifty-five-year-old woman had worried about dyspnea for three years. A chest x-ray appeared normal except for hypovascularity of the upper lung field. On angiograms, a wedge-shaped obstruction was observed at the left proximal pulmonary artery, and well-developed collateral circulation from the right thoracic artery to the internal costal arteries was observed. On the exercise test, exertional dyspnea developed with tachycardia and a decreased saturation of arterial oxygen.


Prediction of peripheral neuropathy in multiple myeloma patients receiving bortezomib and thalidomide: a genetic study based on a single nucleotide polymorphism array.
Bortezomib- and thalidomide-based therapies have significantly contributed to improved survival of multiple myeloma (MM) patients. However, treatment-induced peripheral neuropathy (TiPN) is a common adverse event associated with them. Risk factors for TiPN in MM patients include advanced age, prior neuropathy, and other drugs, but there are conflicting results about the role of genetics in predicting the risk of TiPN. Thus, we carried out a genome-wide association study based on more than 300 000 exome single nucleotide polymorphisms in 172 MM patients receiving therapy involving bortezomib and thalidomide. We compared patients developing and not developing TiPN under similar treatment conditions (GEM05MAS65, NCT00443235). The highest-ranking single nucleotide polymorphism was rs45443101, located in the PLCG2 gene, but no significant differences were found after multiple comparison correction (adjusted P = .1708). Prediction analyses, cytoband enrichment, and pathway analyses were also performed, but none yielded any significant findings. A copy number approach was also explored, but this gave no significant results either. In summary, our study did not find a consistent genetic component associated with TiPN under bortezomib and thalidomide therapies that could be used for prediction, which makes clinical judgment essential in the practical management of MM treatment.


Oxidative stress in human aging and mitochondrial disease-consequences of defective mitochondrial respiration and impaired antioxidant enzyme system.
Respiratory function of mitochondria is compromised in aging human tissues and severely impaired in the patients with mitochondrial disease. A wide spectrum of mitochondrial DNA (mtDNA) mutations has been established to associate with mitochondrial diseases. Some of these mtDNA mutations also occur in various human tissues in an age-dependent manner. These mtDNA mutations cause defects in the respiratory chain due to impairment of the gene expression and structure of respiratory chain polypeptides that are encoded by the mitochondrial genome. Since defective mitochondria generate more reactive oxygen species (ROS) such as O2- and H2O2 via electron leak, we hypothesized that oxidative stress is a contributory factor for aging and mitochondrial disease. This hypothesis has been supported by the findings that oxidative stress and oxidative damage in tissues and culture cells are increased in elderly subjects and patients with mitochondrial diseases. Another line of supporting evidence is our recent finding that the enzyme activities of Cu,Zn-SOD, catalase and glutathione peroxidase (GPx) decrease with age in skin fibroblasts. By contrast, Mn-SOD activity increases up to 65 years of age and then slightly declines thereafter. On the other hand, we observed that the RNA, protein and activity levels of Mn-SOD are increased two- to three-fold in skin fibroblasts of the patients with CPEO syndrome but are dramatically decreased in patients with MELAS or MERRF syndrome. However, the other antioxidant enzymes did not change in the same manner. The imbalance in the expression of these antioxidant enzymes indicates that the production of ROS is in excess of their removal, which in turn may elicit an elevation of oxidative stress in the fibroblasts. Indeed, it was found that intracellular levels of H2O2 and oxidative damage to DNA and lipids in skin fibroblasts from elderly subjects or patients with mitochondrial diseases are significantly increased as compared to those of age-matched controls. Furthermore, Mn-SOD or GPx-1 gene knockout mice were found to display neurological disorders and enhanced oxidative damage similar to those observed in the patients with mitochondrial disease. These observations are reviewed in this article to support that oxidative stress elicited by defective respiratory function and impaired antioxidant enzyme system plays a key role in the pathophysiology of mitochondrial disease and human aging.


Nitric oxide and peroxynitrite. The ugly, the uglier and the not so good: a personal view of recent controversies.
Nitric oxide, a gaseous free radical, is poorly reactive with most biomolecules but highly reactive with other free radicals. Its ability to scavenge peroxyl and other damaging radicals may make it an important antioxidant in vivo, particularly in the cardiovascular system, although this ability has been somewhat eclipsed in the literature by a focus on the toxicity of peroxynitrite, generated by reaction of O2*- with NO* (or of NO- with O2). On balance, experimental and theoretical data support the view that ONOO- can lead to hydroxyl radical (OH*) generation at pH 7.4, but it seems unlikely that OH* contributes much to the cytotoxicity of ONOO-. The cytotoxicity of ONOO- may have been over-emphasized: its formation and rapid reaction with antioxidants may provide a mechanism of using NO* to dispose of excess O2*-, or even of using O2*- to dispose of excess NO*, in order to maintain the correct balance between these radicals in vivo. Injection or instillation of "bolus" ONOO- into animals has produced tissue injury, however, although more experiments generating ONOO- at steady rates in vivo are required. The presence of 3-nitrotyrosine in tissues is still frequently taken as evidence of ONOO- generation in vivo, but abundant evidence now exists to support the view that it is a biomarker of several "reactive nitrogen species". Another under-addressed problem is the reliability of assays used to detect and measure 3-nitrotyrosine in tissues and body fluids: immunostaining results vary between laboratories and simple HPLC methods are susceptible to artefacts. Exposure of biological material to low pH (e.g. during acidic hydrolysis to liberate nitrotyrosine from proteins) or to H2O2 might cause artefactual generation of nitrotyrosine from NO2- in the samples. This may be the origin of some of the very large values for tissue nitrotyrosine levels quoted in the literature. Nitrous acid causes not only tyrosine nitration but also DNA base deamination at low pH: these events are relevant to the human stomach since saliva and many foods are rich in nitrite. Several plant phenolics inhibit nitration and deamination in vitro, an effect that could conceivably contribute to their protective effects against gastric cancer development.


Occurrence of enterotoxigenic Aeromonas species in foods.
Out of a total of 246 food samples of animal origin screened for isolation of Aeromonas spp., 33 (13.41%) were positive for these organisms. Maximum positivity was shown by the samples from fish (28.57%), followed by poultry meat (16.67%), poultry eggs (12.50%), goat meat (12%), buffalo meat (7.69%) and cow milk (5.56%). A. hydrophila was the predominant species (51.52%) followed by A. sobria (39.39%) and A. caviae. Of these, 70.59% A. hydrophila, 69.23% A. sobria and 33.33% A. caviae showed enterotoxigenic reaction in mouse paw oedema test.


The Past, the Present, and the Future of the Size Exclusion Chromatography in Extracellular Vesicles Separation.
Extracellular vesicles (EVs) are cell-derived membranous particles secreted by all cell types (including virus infected and uninfected cells) into the extracellular milieu. EVs carry, protect, and transport a wide array of bioactive cargoes to recipient/target cells. EVs regulate physiological and pathophysiological processes in recipient cells and are important in therapeutics/drug delivery. Despite these great attributes of EVs, an efficient protocol for EV separation from biofluids is lacking. Numerous techniques have been adapted for the separation of EVs with size exclusion chromatography (SEC)-based methods being the most promising. Here, we review the SEC protocols used for EV separation, and discuss opportunities for significant improvements, such as the development of novel particle purification liquid chromatography (PPLC) system capable of tandem purification and characterization of biological and synthetic particles with near-single vesicle resolution. Finally, we identify future perspectives and current issues to make PPLC a tool capable of providing a unified, automated, adaptable, yet simple and affordable particle separation resource.


The extended Granger causality analysis for Hodgkin-Huxley neuronal models.
How to extract directions of information flow in dynamical systems based on empirical data remains a key challenge. The Granger causality (GC) analysis has been identified as a powerful method to achieve this capability. However, the framework of the GC theory requires that the dynamics of the investigated system can be statistically linearized; i.e., the dynamics can be effectively modeled by linear regressive processes. Under such conditions, the causal connectivity can be directly mapped to the structural connectivity that mediates physical interactions within the system. However, for nonlinear dynamical systems such as the Hodgkin-Huxley (HH) neuronal circuit, the validity of the GC analysis has yet been addressed; namely, whether the constructed causal connectivity is still identical to the synaptic connectivity between neurons remains unknown. In this work, we apply the nonlinear extension of the GC analysis, i.e., the extended GC analysis, to the voltage time series obtained by evolving the HH neuronal network. In addition, we add a certain amount of measurement or observational noise to the time series to take into account the realistic situation in data acquisition in the experiment. Our numerical results indicate that the causal connectivity obtained through the extended GC analysis is consistent with the underlying synaptic connectivity of the system. This consistency is also insensitive to dynamical regimes, e.g., a chaotic or non-chaotic regime. Since the extended GC analysis could in principle be applied to any nonlinear dynamical system as long as its attractor is low dimensional, our results may potentially be extended to the GC analysis in other settings.


Kremen proteins are Dickkopf receptors that regulate Wnt/beta-catenin signalling.
The Wnt family of secreted glycoproteins mediate cell cell interactions during cell growth and differentiation in both embryos and adults. Canonical Wnt signalling by way of the beta-catenin pathway is transduced by two receptor families. Frizzled proteins and lipoprotein-receptor-related proteins 5 and 6 (LRP5/6) bind Wnts and transmit their signal by stabilizing intracellular beta-catenin. Wnt/beta-catenin signalling is inhibited by the secreted protein Dickkopf1 (Dkk1), a member of a multigene family, which induces head formation in amphibian embryos. Dkk1 has been shown to inhibit Wnt signalling by binding to and antagonizing LRP5/6. Here we show that the transmembrane proteins Kremen1 and Kremen2 are high-affinity Dkk1 receptors that functionally cooperate with Dkk1 to block Wnt/beta-catenin signalling. Kremen2 forms a ternary complex with Dkk1 and LRP6, and induces rapid endocytosis and removal of the Wnt receptor LRP6 from the plasma membrane. The results indicate that Kremen1 and Kremen2 are components of a membrane complex modulating canonical Wnt signalling through LRP6 in vertebrates.