Prevalence of Eosinophilic Granulomatosis With Polyangiitis and Associated Health Care Utilization Among Patients With Concomitant Asthma in US Commercial Claims Database.
To estimate the prevalence and associated disease burden of eosinophilic granulomatosis with polyangiitis (EGPA) in patients with asthma from a US claims database. Two cohorts were defined using enrollees (aged ≥18 years) from the Optum deidentified Clinformatics Datamart claims database 2010-2014, based on validated EGPA case definitions with varying specificity: EGPA 1 (main cohort; more specific; patients with 2 codes [in any combination] within 12 months of each other for eosinophilia, vasculitis, or mononeuritis multiplex) and EGPA 2 (sensitivity analysis cohort; less specific; patients with 2 codes of above conditions and/or neurologic symptoms within 12 months of each other). Patients had 3 or more asthma medications in the 12-month baseline before index date (date of the second code). Eosinophilic granulomatosis with polyangiitis prevalence, asthma severity during the baseline period, oral corticosteroid (OCS) use, and health care utilization during the 12-month follow-up period were determined. Overall, 88 and 604 patients were included in main cohort EGPA 1 and sensitivity analysis cohort EGPA 2, respectively; corresponding annual EGPA prevalence rates were 3.2 to 5.9 and 23.4 to 30.7 cases/million patients. Approximately 75% of patients were prescribed OCS and ~30% experienced 1 or more hospitalization; 75% in EGPA 1 and 52% in EGPA 2 with 1 or more non-OCS prescription in the 90 days before index date had severe asthma. Eosinophilic granulomatosis with polyangiitis prevalence estimates varied based on specificity of the case definition but were generally consistent with previous country-specific estimates. Despite differences in prevalence, both cohorts displayed a generally similar, high burden of OCS use and health care utilization, highlighting the substantial disease burden among patients with EGPA and the need for specific treatments.


[Danazol action on 5'-nucleotidase activity in mouse peritoneal macrophages].
The paper reports the results of danazol action in vivo and in vitro on 5'-nucleotidase activity of mouse peritoneal macrophages. The results showed, that danazol at a concentration of 10 M (the pharmacologic level in women taking 600 mg/day) significantly stimulated the enzyme activity.


Socio-cultural factors favoring home delivery in Afar pastoral community, northeast Ethiopia: A Qualitative Study.
Despite expanding the number of health facilities, Ethiopia has still the highest home delivery services utilization. Health care service utilization varies between regions within the country. This study explored the socio-cultural factors influencing health facility delivery in a pastoralist region of Afar, Ethiopia. An explorative qualitative study was conducted in October-December 2015. A total of 18 focus group discussions were conducted separately with mothers, male tribal leaders and religious leaders. In addition, 24 key informant interviews were conducted with Women's Affairs Bureau and district health office experts and traditional birth attendants and all were selected purposively. Data were coded and categorized using open code software and analyzed based on a thematic approach. The social factors that affect the choice of delivery place include workload, lack of independence and decision-making power of women, and lack of substitute for childcare and household chores during pregnancy and childbirth. The cultural and spiritual factors include assuming delivery as natural process ought to happen at home, trust in traditional birth attendants, traditional practices during and after delivery and faithful to religion practice, besides, denial by health facilities to benign traditional and spiritual practices such as prayers and traditional food preparations to be performed over there. Socio-cultural factors are far more than access to health centers as barriers to the utilization of health facilities for child birth. The provision of a maternity waiting home around the health facilities can alleviate some of these socio-cultural barriers.


Relation between myocardial infarct location and stroke.
We sought to compare the likelihood of stroke in patients with anterior versus nonanterior myocardial infarction. The association between anterior infarction and left ventricular thrombus has led to the assumption that embolization from thrombi is an important cause of stroke in patients with anterior infarction. We hypothesized that if anterior infarction is a cause of left ventricular thrombi, the number of strokes should be disproportionately higher in patients with anterior than nonanterior infarction. We performed a retrospective analysis of 2,466 patients randomized from day 3 to day 15 after infarction as part of a multicenter placebo-controlled study of diltiazem to prevent cardiac death or myocardial infarction. Any acute focal cerebral disorder resulting in localizing findings characterized as a stroke or transient ischemic attack was considered an event. Of 91 events during a follow-up period of 12 to 52 months, 23 (3.2%) occurred in 724 patients with an anterior and 68 (3.9%) in 1,742 patients with a nonanterior myocardial infarction (relative risk 0.81; 95% confidence interval 0.51 to 1.30). Power analysis revealed that the negativity of the study was not the result of inadequate sample size. Life table analysis showed no difference in cumulative event rate (p = 0.42) according to site of infarction. Cox regression analysis showed that of 10 clinical covariates, only systolic blood pressure was predictive of stroke (p < 0.001). The use of warfarin did not contribute to the model. Finally, the addition of site of infarction (anterior vs. nonanterior) did not contribute significantly to the Cox model. Although there is a significant incidence of stroke after acute myocardial infarction, there is no relation between the occurrence of stroke and site of infarction. These data do not support the presumed causal relation between anterior myocardial infarction, thrombus and stroke.


Assessment of immunogenicity and safety following primary and booster immunisation with a CRM197 -conjugated Haemophilus influenzae type B vaccine in healthy Chinese infants.
Invasive meningitis and pneumonia caused by Haemophilus influenzae type b (Hib) is an important cause of childhood mortality in countries where Hib vaccination is not routine. We evaluated the non-inferiority of a licensed Hib vaccine, PRP-CRM(197) compared with a second licensed Hib vaccine, PRP-T, following the recommended Chinese immunisation schedule for infants between 6 months and 1 year of age. In the first study phase, 6-12 month-old infants received two primary doses of either PRP-CRM(197) (n = 335) or PRP-T (n = 335) vaccine administered 1 month apart. In the second study phase 8 months later, the same children received a single booster dose of vaccine identical to that use for priming (PRP-CRM(197), n = 327; PRP-T, n = 333). Serum levels of anti-polyribosylribitol phosphate (PRP) antibodies were measured using enzyme-linked immunosorbent assay (ELISA). Non-inferiority of primary and booster doses was assessed in terms of percentages of subjects with anti-PRP antibody levels associated with providing short-term (≥ 0.15 μg/ml) and long-term (≥ 1.0 μg/ml) protection; the non-inferiority margin was set at -5%. PRP-CRM(197) was demonstrated to be non-inferior to PRP-T. Anti-PRP antibodies levels ≥ 0.15 μg/ml and ≥ 1.0 μg/ml were achieved by 97% of infants in the PRP-CRM(197) group and 98% of infants in the PRP-T group 1 month after primary immunisation, and by all subjects (100%) in both vaccine groups 1 month after booster administration. Safety profiles for both vaccines were similar; no serious adverse events, deaths or adverse events leading to withdrawal occurred during the study. PRP-CRM(197) was well-tolerated and immunologically non-inferior to a licensed comparator Hib vaccine in Chinese infants (Clinicaltrials.gov: NCT01044316 & NCT01226953).


Potentially modifiable predictors of mortality in patients treated with long-term oxygen therapy.
Anemia is considered a systemic manifestation of Chronic Obstructive Pulmonary Disease (COPD); however, few studies have described its influence on chronic respiratory failure (CRF) prognosis. We aimed to test the hypotheses that anemia negatively influences survival and also to identify the cut-off points of hematocrit (Htc) and hemoglobin (Hb) associated with higher mortality in CRF patients using long-term oxygen therapy (LTOT). One-hundred forty two patients with CRF in use of LTOT were evaluated at baseline and followed for three years or until death. Baseline assessment included identification, diagnosis, body composition, dyspnea, health status (HS), spirometry, arterial blood gases, Hb and Htc. Univariate and Cox proportional hazard models were used to evaluate predictors of mortality. We performed ROC curve to identify the best cut-off point of the variables related to survival to construct the Kaplan-Meier curves. Eight-three patients (58%) died after three years. Baseline values of Hb and Htc were significantly lower in the non-survivors group and both, Htc (HR, 0.96; 95%CI 0.91-0.99; p = 0.04), Hb (HR, 0.86; 95%CI 0.76-0.98; p = 0.02) were selected as predictors of mortality after three years. The cut-off points determined were: the value of HB is < 11 g/dl (sensitivity 95% specificity 85%), Htc ≤ 33% (sensitivity 97% specificity 89%). Other prognostic factors were: male gender, low PaCO(2) and SpO(2), higher dyspnea perception and impairment of HS. Our study shows that anemia is a predictor of mortality in patients with CRF under LTOT treatment. Although anemia is potentially modifiable, the effects of raising hemoglobin on mortality remain undetermined.


Treatment of severe infrapopliteal vascular disease using a novel atherectomy device: a case report.
We report two first in man cases of percutaneous revascularization for severe intrapopliteal arterial disease using the novel PV atherectomy device.


The vulvar clinic: an eight-year appraisal.
One thousand forty-three patients made 2,750 visits to the Vulvar Clinic at the Milwaukee County Medical Complex from July, 1970, to July, 1978. This experience is analyzed and the various diseases are grouped in order of frequency. The results shed light on those vulvar problems most likely to be encountered in general office practice and point the way toward future areas of research.


Gingival biotype modification with collagen matrix or autogenous subepithelial connective tissue graft: Histologic and volumetric analyses in a beagle model.
To evaluate the volumetric effect and biocompatibility of porcine tendon-derived type I collagen matrix graft (CG) in gingival biotype modification (GBM) compared with subepithelial connective tissue graft (SCTG) in a beagle model. Surface analysis using scanning electron microscopy and a collagen degradation assay of CG was performed in vitro. Six adult dogs were used in in vivo experiment, and each received autologous SCTG or CG at the anterior side. Histometric and three-dimensional digital volume analyses were conducted to compare quantitative changes in CG and SCTG in GBM. Immunohistochemical analysis was performed for the qualitative evaluation of CG compared to SCTG. CG had a double-layered structure, and its degradation was slower than that of other well-reported materials. No critical problems were associated with the healing procedure. Changes in gingival thickness and volume in the CG and SCTG groups were equivalent, with no significant differences between the groups. Type I collagen and vascular endothelial growth factor expression levels were similar in both groups. CG and SCTG had equivalent potential for GBM in terms of quantity and quality. Additionally, CG could be used as a reasonable substitute for SCTG, making surgery convenient and predicting successful clinical outcomes.


Treating impaired cognition in schizophrenia: the case for combining cognitive-enhancing drugs with cognitive remediation.
Cognitive impairment is a well-documented feature of schizophrenia and represents a major impediment to the functional recovery of patients. The therapeutic strategies to improve cognition in schizophrenia have either used medications (collectively referred to as 'cognitive-enhancing drugs' in this article) or non-pharmacological training approaches ('cognitive remediation'). Cognitive-enhancing drugs have not as yet been successful and cognitive remediation has shown modest success. Therefore, we may need to explore new therapeutic paradigms to improve cognition in schizophrenia. The optimal approach may require a combination of cognitive-enhancing drugs with cognitive remediation. We review the available data from animal and human studies that provide the conceptual basis, proof-of-concept and illustrations of success of such combination strategies in experimental and clinical paradigms in other conditions. We address the major design issues relevant to the choice of the cognitive-enhancing drugs and cognitive remediation, as well as the timing and the duration of the intervention as will be relevant for schizophrenia. Finally, we address the practical realities of the development and testing of such combined approaches in the real-world clinical situation and conclude that while scientifically attractive, there are several practical difficulties to be overcome for this approach to be clinically feasible.


Comparative genomics of Sphingopyxis spp. unravelled functional attributes.
Members of genus Sphingopyxis are known to thrive in diverse environments. Genomes of 21 Sphingopyxis strains were selected. Phylogenetic analysis was performed using GGDC, AAI and core-SNP showed agreement at sub-species level. Based on our results, we propose that both S. baekryungensis DSM16222 and Sphingopyxis sp. LPB0140 strains should not be included under genus Sphingopyxis. Core-analysis revealed, 1422 genes were shared which included essential pathways and genes for conferring adaptation against stress environment. Polyhydroxybutyrate degradation, anaerobic respiration, type IV secretion were notable abundant pathways and exopolysaccharide, hyaluronic acid production and toxin-antitoxin system were differentially present families. Interestingly, genome of S. witflariensis DSM14551, Sphingopyxis sp. MG and Sphingopyxis sp. FD7 provided a hint of probable pathogenic abilities. Protein-Protein Interactome depicted that membrane proteins and stress response has close integration with core-proteins while aromatic compounds degradation and virulence ability formed a separate network. Thus, these should be considered as strain specific attributes.


Aminoborylation/Suzuki-Miyaura tandem cross coupling of aryl iodides as efficient and selective synthesis of unsymmetrical biaryls.
Sequential borylation of a first aryl iodide using a dialkylaminoborane followed by a Suzuki-Miyaura cross coupling of second aryl iodide ended up with an efficient, selective and practical synthesis of unsymmetrical biaryls. This tandem coupling shows a wide range of applicability.


Selective 19F MR imaging of 5-fluorouracil and alpha-fluoro-beta-alanine.
A 19F MR chemical shift imaging (CSI) technique is presented which enables selective imaging of the antineoplastic drug 5-fluorouracil (5-FU) and its major catabolite alpha-fluoro-beta-alanine (FBAL). The CSI sequence employs a chemical shift selective (CHESS) saturation pulse to suppress either the 5-FU or the FBAL resonance before the other component of the two-line 19F MR spectrum is measured. Because the transmitter frequency can always be set to the Larmor frequency of the 19F resonance to be imaged, this approach yields 5-FU and FBAL MR images free of chemical shift artifacts in read-out and slice-selection direction. In phantom experiments, selective 5-FU and FBAL images with a spatial resolution of 15 x 15 x 20 mm3 (4.5 ml) were obtained in 30 min from a model solution, whose drug and catabolite concentrations were similar to those estimated in the liver of tumor patients undergoing IV chemotherapy with 5-FU. The drug-specific MR imaging technique developed is, therefore, well-suited for the direct and noninvasive monitoring of the up-take and trapping of 5-FU in liver tumors in vivo.


Female Sexual Function in Young Women With Type 1 Diabetes and Additional Autoimmune Diseases.
Female sexual dysfunctions (FSDs) are frequent concerns in women with type 1 diabetes (T1D), which is frequently associated with other autoimmune diseases (ADs). To assess sexual function in young type 1 diabetic women with or without additional ADs. Women with T1D aged 18-35 years with a stable couple relationship and no oral contraceptive use were enrolled. Diabetic women with concomitant ADs were also identified. All women completed the Female Sexual Function Index (FSFI) and the Female Sexual Distress Scale. The main outcome was the prevalence of FSD. The FSFI-single domain scores were also evaluated in diabetic women with or without additional ADs. The global population included 154 diabetic women, of whom 66 (42%) had at least one additional AD. The prevalence of FSD was similar among diabetic women with and those without (30% vs 32%, P = .980) additional ADs. The FSFI-desire score was significantly lower among diabetic women with concomitant ADs than those without ADs [median (interquartile range), 4.1 (3.6, 4.8) vs 4.6 (4.0, 5.0), P = .042]. An early evaluation of sexual function in women with T1D and concomitant ADs should be encouraged. Major strengths are the use of 2 validated tools to diagnose FSD and the relatively large number of subjects investigated. The limitations include the cross-sectional nature of the study, which does not allow to make inference regarding the cause and effect. Diabetic women with additional ADs show an impairment in sexual desire as compared with those suffering only from diabetes. Longo M, Cirillo P, Scappaticcio L, et al. Female Sexual Function in Young Women With Type 1 Diabetes and Additional Autoimmune Diseases. J Sex Med 2021;18:219-223.


Untangling the effects of hunger, anxiety, and nausea on energy intake during intravenous cholecystokinin octapeptide (CCK-8) infusion.
Infusions of cholecystokinin (CCK) may exert their effects on appetite by inducing feelings of nausea or anxiety. In this double blind, placebo controlled crossover study, the impact of these effects on appetite were examined. Fifteen male subjects received a 20 min i.v. infusion of cholecystokinin octapeptide (CCK-8) (4 ng/kg/min) or saline. The infusion commenced 20 min after a soup preload and 10 min before an ad libitum test meal. Visual analogue scales of appetite and mood were measured over 3 h, and subjects were instructed to report any other sensations they experienced over this time. CCK-8 significantly reduced premeal hunger, elevated premeal anxiety, and reduced energy intake at the ad libitum test meal. Meal duration and rate of eating (kcal/min) were also significantly reduced after CCK-8. After the smaller meal with CCK-8, hunger rose quickly to a higher level than with placebo. The return of hunger was commensurate with the smaller amount of energy consumed and indicated that CCK did not exert an enduring effect on hunger suppression. A significant correlation was found between the reduction in energy intake and hunger (r = 0.75 p < 0.01), but not with anxiety (r = 0.15 not significant). Analyses were performed separately on subjects who did (n = 8), or did not (n = 7) report gastrointestinal disturbance. Energy intake was reduced by 56.6% and 44.6%, respectively. These results indicate that, although feelings of anxiety and nausea may accompany CCK infusions, they are not necessary for the effects of CCK on appetite. These data provide support for a role of CCK in satiety.


Nutlin-3 acts as a DNA methyltransferase inhibitor to sensitize esophageal cancer to chemoradiation.
Esophageal squamous cell carcinoma (ESCC) is highly resistant to chemoradiation therapy. We aimed to examine whether Nutlin-3, a molecule that suppresses murine double min 2 (MDM2)-mediated p53 and Retinoblastoma (RB) protein degradation leading to downregulation of DNA methyltransferases (DNMTs), can be a novel therapeutic agent for ESCC. We used wild-type and chemoradiation-resistant ESCC cell lines in this study. The expression of DNMTs, p53 and RB, and methylation level of tumor suppressor genes (TSG) were analyzed upon Nutlin-3 treatment. The antitumor efficacy of Nutlin-3 was investigated in ESCC cell lines and xenograft tumor model. TSG protein expression was checked in the excised tumor tissue. Nutlin-3 induced upregulation of p53 and RB and downregulation of DNMTs proteins in the chemoradiation-resistant and aggressive ESCC cells. The methylation level of TSGs was decreased by Nutlin-3. Nutlin-3 inhibits clonogenic growth of ESCC cells and exerts a synergistic cytotoxic-effect when combined with chemotherapeutic agent cisplatin. Moreover, xenograft tumor growth in SCID mice was suppressed by Nutlin-3. The protein expression level of DNMTs was downregulated, and that of TSGs was upregulated by Nutlin-3 treatment in the excised tumor tissue. In conclusion, Nutlin-3 is a potential therapeutic agent that can potentiate the treatment efficacy of chemoradiation-resistant ESCC.


Disruption in neuropeptide Y and leptin signaling in obese ventromedial hypothalamic-lesioned rats.
Electrolytic lesions placed in the ventromedial hypothalamus (VMH) of rats induce instant hyperphagia and excessive weight gain. Since neuropeptide Y (NPY) is a potent hypothalamic orexigenic signal, and leptin secreted by adipocytes regulates NPY output, we tested the hypothesis that altered NPYergic-leptin signaling may underlie hyperphagia in VMH-lesioned rats. VMH-lesioned rats exhibiting hyperphagia and excessive weight gain in a time-related fashion were sacrificed on days 2, 7, and 21 post-surgery. Quite unexpectedly, NPY concentrations in the hypothalamic paraventricular nucleus (PVN), a major site of NPY release for stimulation of feeding, and in other sites, such as the dorsomedial nucleus, lateral hypothalamic area and median eminence-arcuate nucleus decreased, with the earliest diminution occurring on day 2 in the PVN only. In vitro basal and K+-evoked NPY release from the PVN of VMH-lesioned rats was significantly lower than that of controls. Analysis of hypothalamic NPY gene expression showed that although the daily decrease in NPY mRNA from 0800 to 2200 h occurred as in control rats, NPY mRNA concentrations were markedly reduced at these times in the hypothalami of VMH-lesioned rats. Leptin synthesis in adipocytes as indicated by leptin mRNA levels was also profoundly altered in VMH-lesioned rats. The daily pattern of increase in adipocyte leptin mRNA at 2200 h from 0800 h seen in controls was abolished, higher levels of leptin gene expression at 2200 h were maintained at 0800 h. The pattern of increase in serum leptin and insulin levels diverged in VMH-lesioned rats. Serum insulin concentration increased to maximal on day 2 and remained at that level on day 21-post-lesion; serum leptin levels on the other hand, increased slowly in a time-related fashion during this period. These results demonstrate that hyperphagia and excessive weight gain in VMH-lesioned rats are associated with an overall decrease in hypothalamic NPY and augmented leptin signaling to the hypothalamus. The divergent time course of increases in serum leptin and insulin levels suggest independent mechanisms responsible for their augmented secretion, and neither these hormones nor VMH lesions altered the daily rhythm in NPY gene expression. These observations underscore the existence of an independent mechanism controlling the daily rhythm in hypothalamic NPY gene expression and suggest that leptin feedback action requires an intact VMH.


UPLC-TOF-MS for plant metabolomics: a sequential approach for wound marker analysis in Arabidopsis thaliana.
The model plant Arabidopsis thaliana was studied for the search of new metabolites involved in wound signalling. Diverse LC approaches were considered in terms of efficiency and analysis time and a 7-min gradient on a UPLC-TOF-MS system with a short column was chosen for metabolite fingerprinting. This screening step was designed to allow the comparison of a high number of samples over a wide range of time points after stress induction in positive and negative ionisation modes. Thanks to data treatment, clear discrimination was obtained, providing lists of potential stress-induced ions. In a second step, the fingerprinting conditions were transferred to longer column, providing a higher peak capacity able to demonstrate the presence of isomers among the highlighted compounds.


PPAR-γ promotes p38 MAP kinase-mediated endothelial cell permeability through activating Sirt3.
Ischemia-reperfusion (I/R)-induced vascular dysfunction is the main factor to acute ischemic stroke. Sirt3 is one of the sirtuin family members, which plays an important role in the development of neurological diseases. In this study, we constructed I/R injury model on HBMEC cells and induced the overexpression of Sirt3 in model cells. Meanwhile, the p38 activator U-46619 was used to examine the connection between Sirt3 and p38. We also examined the level of endothelial associated proteins, including occluding, ZO-1 and claudin-4 by using qRT-PCR and western blot. Our findings indicated that overexpression of Sirt3 decreased the permeability of model cells and promoted in the growth of endothelial cells. However, the activation of p38 could antagonize the function of Sirt3 in HBMEC cells. Moreover, Our results indicated a positive correlation between Sirt3 and inter-endothelial junction proteins. Importantly, PPAR-γ agonist and inhibitor were utilized to investigate the role of PPAR-γ in Sirt3 mediated cell function. Sirt3 was targeted by PPAR-γ in model cells. Taken together, this research not only demonstrated PPAR-γ might benefit to the growth of endothelial cell though activating Sirt3 but also indicated its potential value in the treatment for ischemic stroke.


Allometric analysis of the atrioventricular heart-valve weights in beagle dogs.
A statistical analysis of atrioventricular heart valve weights of 218 beagle dogs was performed. Applying the allometric function, the correlation between body or heart weight and the weights of the mitral valve, tricuspid valve-angular/parietal leaflet and tricuspid valve-septal leaflet could be described statistically. In the age range between 2 and 81 weeks a significant linear, positive correlation was found for all three parts. The mitral valve and the angular and parietal leaflet of the tricuspid valve show a negative allometry or slower growth rate than body or heart, whereas the septal leaflet grows isometrically. Sex differences were not found. The relevance of such data for the evaluation of certain pathological findings is discussed.


Changes in HIV treatment differentiated care uptake during the COVID-19 pandemic in Zambia: interrupted time series analysis.
Differentiated service delivery (DSD) models aim to improve the access of human immunodeficiency virus treatment on clients and reduce requirements for facility visits by extending dispensing intervals. With the advent of the COVID-19 pandemic, minimising client contact with healthcare facilities and other clients, while maintaining treatment continuity and avoiding loss to care, has become more urgent, resulting in efforts to increase DSD uptake. We assessed the extent to which DSD coverage and antiretroviral treatment (ART) dispensing intervals have changed during the COVID-19 pandemic in Zambia. We used client data from Zambia's electronic medical record system (SmartCare) for 737 health facilities, representing about three-fourths of all ART clients nationally. We compared the numbers and proportional distributions of clients enrolled in DSD models in the 6 months before and 6 months after the first case of COVID-19 was diagnosed in Zambia in March 2020. Segmented linear regression was used to determine whether the outbreak of COVID-19 in Zambia further accelerated the increase in DSD scale-up. Between September 2019 and August 2020, 181,317 clients aged 15 or older (81,520 and 99,797 from 1 September 2019 to 1 March 2020 and from 1 March to 31 August 2020, respectively) enrolled in DSD models in Zambia. Overall participation in all DSD models increased over the study period, but uptake varied by model. The rate of acceleration increased in the second period for home ART delivery (152%), ≤ 2-month fast-track (143%) and 3-month MMD (139%). There was a significant reduction in the enrolment rates for 4- to 6-month fast-track (-28%) and "other" models (-19%). Participation in DSD models for stable ART clients in Zambia increased after the advent of COVID-19, but dispensing intervals diminished. Eliminating obstacles to longer dispensing intervals, including those related to supply chain management, should be prioritized to achieve the expected benefits of DSD models and minimize COVID-19 risk.


Vaccination and immunotherapies in neuroimmunological diseases.
Neuroimmunological diseases and their treatment compromise the immune system, thereby increasing the risk of infections and serious illness. Consequently, vaccinations to protect against infections are an important part of the clinical management of these diseases. However, the wide variety of immunotherapies that are currently used to treat neuroimmunological disease - particularly multiple sclerosis and neuromyelitis optica spectrum disorders - can also impair immunological responses to vaccinations. In this Review, we discuss what is known about the effects of various immunotherapies on immunological responses to vaccines and what these effects mean for the safe and effective use of vaccines in patients with a neuroimmunological disease. The success of vaccination in patients receiving immunotherapy largely depends on the specific mode of action of the immunotherapy. To minimize the risk of infection when using immunotherapy, assessment of immune status and exclusion of underlying chronic infections before initiation of therapy are essential. Selection of the required vaccinations and leaving appropriate time intervals between vaccination and administration of immunotherapy can help to safeguard patients. We also discuss the rapidly evolving knowledge of how immunotherapies affect responses to SARS-CoV-2 vaccines and how these effects should influence the management of patients on these therapies during the COVID-19 pandemic.


Were self-described introverts "immune" to increased drug use and entrapment during the pandemic?
Social restrictions and other stressors related to the Covid-19 pandemic disrupted daily life in ways that might have increased drug use and undermined mental health. We investigated whether such changes depended on the amount and quality of a person's social activity. We also evaluated the popular idea that effects of pandemic-related restrictions would depend on introversion; to this end, we used self-described introversion as a proxy for preferred frequency of social activity. Between September 2020 and March 2021, we obtained online-survey data from 2615 respondents who retrospectively reported alcohol, opioid, or psychostimulant use. We analyzed (1) changes in drug use and entrapment (a psychological construct linked to suicidality) as a function of introversion and the frequency and quality of social activity, and (2) changes in drug use as a function of change in entrapment. Most felt more entrapped during the pandemic, but only a minority increased drug use. Generally: (1) entrapment and drug use increased in respondents unsatisfied with their social activity, (2) introversion and frequency of activity had less influence than satisfaction, (3) introverts reported more symptoms of entrapment, anxiety, depression, and loneliness than non-introverts, (4) when social activity was frequent and unsatisfying, psychostimulant use increased in introverts and opioid use increased in extraverts, (5) alcohol use increased in those who felt increased entrapment, and (6) alcohol and opioid use decreased in those who felt decreased entrapment. Satisfactory social activity (even in small amounts) was associated with better outcomes, mostly without regard to introversion.


Reproductive effort in clonal plants: constant allocation ratios among ramets?
Armstrong (1982, 1983) predicted that all ramets within a clone should have the same ratio of biomass allocation to sexual reproduction versus vegetative growth. He presented data (1984) that he interpreted as showing that Solidago altissima ramets in a clone do have the predicted constant allocation ratio. Reanalysis of his methods shows that this conclusion was an artifact of his analysis. A simulation using random numbers and Armstrong's analysis showed the same pattern as his data. Data from S. altissima ramets of a single clone grown in a greenhouse experiment, using a different analysis, illustrated that the allocation ratios within a clone can be highly variable.


Quality of diabetes care in the UK: comparison of published quality-of-care reports with results of the Quality and Outcomes Framework for Diabetes.
To conduct a systematic review of published observational studies of quality of diabetes care in primary care in the UK and to compare the results with the quality of care data from the Quality and Outcomes Framework (QOF) of the new General Practice Contract in the UK. medline and embase were searched for articles published from 1999 to June 2006. We also searched for reference lists of studies that fitted our inclusion criteria. All members of the Primary Care Diabetes Europe were contacted and asked to send lists of any relevant published articles. Abstracts were reviewed and data were collected independently by two authors. Abstracts of 742 papers were identified, of which six papers fulfilled the final selection criteria. The total number of people included in the six published studies was 83 098 (a range of 504 to 54 180 people) compared with the UK QOF data of 1.8 million people with diabetes. The quality indicators for assessment of care varied between different published studies, making comparisons more difficult. Overall, there was a trend towards improvement in both process and outcome of care in the published studies. The quality of care achieved as a result of QOF was greater than that found in published studies. There have been improvements in both process and outcome measures recorded in publications of quality of diabetes care in the UK between 2000 and 2004. Modest financial incentives in primary care are a successful method of improving care for people with diabetes.


Hierarchy of orofacial rhythms revealed through whisking and breathing.
Whisking and sniffing are predominant aspects of exploratory behaviour in rodents. Yet the neural mechanisms that generate and coordinate these and other orofacial motor patterns remain largely uncharacterized. Here we use anatomical, behavioural, electrophysiological and pharmacological tools to show that whisking and sniffing are coordinated by respiratory centres in the ventral medulla. We delineate a distinct region in the ventral medulla that provides rhythmic input to the facial motor neurons that drive protraction of the vibrissae. Neuronal output from this region is reset at each inspiration by direct input from the pre-Bötzinger complex, such that high-frequency sniffing has a one-to-one relationship with whisking, whereas basal respiration is accompanied by intervening whisks that occur between breaths. We conjecture that the respiratory nuclei, which project to other premotor regions for oral and facial control, function as a master clock for behaviours that coordinate with breathing.


Endogenous opiates: 1986.
This is the ninth installment of our annual review of research involving the endogenous opiate peptides. It is restricted to the non-analgesic and behavioral studies of the opiate peptides published in 1986. The specific topics this year include stress; tolerance and dependence; eating; drinking; gastrointestinal, renal, and hepatic processes; mental illness; learning, memory, and reward; cardiovascular responses; respiration and thermoregulation; seizures and other neurological disorders; activity; sex, pregnancy, and development; and some other behaviors.


Biochemical and metabolomic insights into antifungal mechanism of berberine against Candida glabrata.
An unprecedented expansion of antifungal therapy failure incidences in healthcare settings of Candida glabrata is the matter of global concern that needs to be addressed efficiently and effectively. In this pursuit, the present study has investigated the antifungal mechanism of benzylisoquinoline alkaloid berberine using biochemical, metabolic, and gene expression analysis, with the aim to delineate its therapeutic activity against C. glabrata and differentially fluconazole-responsive clinical isolates. Interestingly, the clinical isolates were found to be highly susceptible to berberine. Berberine was found to control the surface properties like hydrophobicity and charge of the cells. The cell membrane composition was altered by berberine, where the ergosterol and fatty acids were affected. The efflux pump activity was inhibited, and osmotic stress was generated in C. glabrata cells upon berberine exposure. The berberine has also generated oxidative stress and activated antioxidant system in C. glabrata cells. Furthermore, these observations were supported by the transcriptional expression study of C. glabrata cell genes (CDR1, RLM1, SLT2, SUR4, KRE1) and metabolomics analysis. Based on fold change analysis, the study identified 20 differential metabolites upon berberine treatment, which belong to central carbon, amino acids, and nucleotide pathways. The checkerboard analysis revealed the potentiation of some classically used antifungal drugs by berberine, thus suggesting it as a combinatorial nutraceutical adjuvant for the eradication of fungal infections. KEY POINTS: • Berberine exhibited better potency against azole-resistant clinical isolates • Berberine modulated metabolites of different pathways • Berberine generated oxidative stress and blocked efflux pump activity.


Stretchable and self-healable hyaluronate-based hydrogels for three-dimensional bioprinting.
Hydrogels have been widely exploited as inks for three-dimensional (3D) bioprinting, a useful technique for building complex biological structures with living cells. However, hydrogels have inherently limited mechanical properties (e.g., brittleness) and printability. Thus, we hypothesized that hyaluronate-based hydrogels with stretchable and self-healing properties would be useful for 3D bioprinting. Oxidized hyaluronate (oHA) and hydrazide-modified hyaluronate (hHA) formed stretchable and flexible hydrogels because of double network formation via chemical cross-linking (i.e., acylhydrazone bond formation) and physical cross-linking (i.e., charge interaction). The addition of adipic acid dihydrazide (ADH) to oHA/hHA hydrogels enhanced the self-healing capability of the gels, which were useful for fabricating 3D constructs with various shapes maintaining their stretchability even after 3D printing (about two times its original length). ATDC5 cells were viable within the 3D-printed constructs in vitro. This hydrogel system, consisting of hyaluronic acid (HA)-based polymers, may have potential for many tissue engineering applications via 3D bioprinting.


Shining a light on SSP4: A comprehensive analysis and biological applications for the detection of sulfane sulfurs.
Fluorescent probes are useful tools for the detection of sulfane sulfurs in biological systems. In this work, we report the development of SSP4, a widely used probe generated in our laboratory. We describe its evolution, preparation, and physical/chemical properties. Fluorescence analyses of SSP4 determined its high selectivity and sensitivity to sulfane sulfurs, even with the interfering presence of other species, such as amino acids and metal ions. Protocols for using SSP4 in a relatively quick and simple manner for the detection of persulfidated proteins, including papain, BSA, and GAPDH were developed. The method was then applied to human protein disulfide isomerase (PDI), leading to the discovery that persulfidation can occur at PDI's non-active site cysteines, and that PDI reductase activity is affected by sulfane sulfur treatment. Protocols for using SSP4 for the bioimaging of exogenous and endogenous sulfane sulfurs in different -cell lines were also established. These results should guide further applications of SSP4.


Impact of Therapeutics on Unified Immunity During Allergic Asthma and Respiratory Infections.
Asthma is a common chronic respiratory disease that affects millions of people worldwide. Patients with allergic asthma, the most prevalent asthma endotype, are widely considered to possess a defective immune response against some respiratory infectious agents, including viruses, bacteria and fungi. Furthermore, respiratory pathogens are associated with asthma development and exacerbations. However, growing data suggest that the immune milieu in allergic asthma may be beneficial during certain respiratory infections. Immunomodulatory asthma treatments, although beneficial, should then be carefully prescribed to avoid misuse and overuse as they can also alter the host microbiome. In this review, we summarize and discuss recent evidence of the correlations between allergic asthma and the most significant respiratory infectious agents that have a role in asthma pathogenesis. We also discuss the implications of current asthma therapeutics beyond symptom prevention.


Gene expression profiling of facilitated L-LTP in VP16-CREB mice reveals that BDNF is critical for the maintenance of LTP and its synaptic capture.
Expression of VP16-CREB, a constitutively active form of CREB, in hippocampal neurons of the CA1 region lowers the threshold for eliciting the late, persistent phase of long-term potentiation (L-LTP) in the Schaffer collateral pathway. This VP16-CREB-mediated L-LTP differs from the conventional late phase of LTP in not being dependent on new transcription. This finding suggests that in the transgenic mice the mRNA transcript(s) encoding the protein(s) necessary for this form of L-LTP might already be present in CA1 neurons in the basal condition. We used high-density oligonucleotide arrays to identify the mRNAs differentially expressed in the hippocampus of transgenic and wild-type mice. We then explored the contribution of the most prominent candidate genes revealed by our screening, namely prodynorphin, BDNF, and MHC class I molecules, to the facilitated LTP of VP16-CREB mice. We found that the overexpression of brain-derived neurotrophic factor accounts for an important component of this phenotype.


Prognostic impact of GATA binding protein-3 expression in primary lung adenocarcinoma.
GATA binding protein-3 (GATA3) is a transcription factor that regulates cell differentiation and maintenance in some types of normal cells. This study aimed to investigate the association between GATA3 expression and primary lung adenocarcinoma and to clarify the clinical significance of GATA3 expression in lung adenocarcinoma. Immunohistochemical GATA3 expression was evaluated using completely resected lung adenocarcinoma samples from 95 cases. GATA3 immunohistochemical staining was performed and scored. Associations between clinicopathological factors and GATA3 expression were analyzed by using the χ2 test and Fisher exact test. The Kaplan-Meier method was used to analyze overall survival (OS) and disease-free survival (DFS). Forty-nine cases expressed high levels of GATA3, which were associated with lymphatic invasion (P=.003). In univariate and multivariate analyses, vascular invasion (P<.001) and high GATA3 expression (P=.023) were identified as independent risk factors for OS. Higher pathological stages (P=.012), vascular invasion (P=.010), and high GATA3 expression (P=.009) were identified as independent risk factors for DFS. The high GATA3 expression group exhibited statistically worse OS (P=.031) and DFS (P=.011) than the low-expression group based on the Kaplan-Meier curves. In resected lung adenocarcinoma, high GATA3 expression is associated with poorer prognosis for both OS and DFS. Therefore, the immunohistochemical evaluation of GATA3 represents a potentially useful prognostic tool for postoperative patients.


Structure-guided design of aminopyrimidine amides as potent, selective inhibitors of lymphocyte specific kinase: synthesis, structure-activity relationships, and inhibition of in vivo T cell activation.
The lymphocyte-specific kinase (Lck), a member of the Src family of cytoplasmic tyrosine kinases, is expressed in T cells and natural killer (NK) cells. Genetic evidence, including knockout mice and human mutations, demonstrates that Lck kinase activity is critical for normal T cell development, activation, and signaling. Selective inhibition of Lck is expected to offer a new therapy for the treatment of T-cell-mediated autoimmune and inflammatory disease. With the aid of X-ray structure-based analysis, aminopyrimidine amides 2 and 3 were designed from aminoquinazolines 1, which had previously been demonstrated to exhibit potent inhibition of Lck and T cell proliferation. In this report, we describe the synthesis and structure-activity relationships of a series of novel aminopyrimidine amides 3 possessing improved cellular potency and selectivity profiles relative to their aminoquinazoline predecessors 1. Orally bioavailable compound 13b inhibited the anti-CD3-induced production of interleukin-2 (IL-2) in mice in a dose-dependent manner (ED 50 = 9.4 mg/kg).


Forthcoming complications in recovered COVID-19 patients with COPD and asthma; possible therapeutic opportunities.
Infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been growing swiftly worldwide. Patients with background chronic pulmonary inflammations such as asthma or chronic obstructive pulmonary diseases (COPD) are likely to be infected with this virus. Of note, there is an argument that COVID-19 can remain with serious complications like fibrosis or other pathological changes in the pulmonary tissue of patients with chronic diseases. Along with conventional medications, regenerative medicine, and cell-based therapy could be alternative approaches to compensate for organ loss or restore injured sites using different stem cell types. Owing to unique differentiation capacity and paracrine activity, these cells can accelerate the healing procedure. In this review article, we have tried to scrutinize different reports related to the harmful effects of SARS-CoV-2 on patients with asthma and COPD, as well as the possible therapeutic effects of stem cells in the alleviation of post-COVID-19 complications. Video abstract.


A novel field method for preserving African lion (Panthera leo) fecal samples for noninvasive hormone metabolite analysis.
The traditional methods of preserving fecal samples to suspend hormone-degrading bacteria are not always options during remote fieldwork when studying wildlife endocrinology. Our goal was to develop a field method for preserving African lion (Panthera leo) feces for hormone metabolite analysis by determining the stability of fecal hormone metabolites: 1) when exposed to the natural environment, and 2) in silica beads at ambient temperatures. We collected fresh feces from zoo-housed lions and combined them into two (male and female) homogenous samples. Each was divided into eight samples to undergo a cross-designed treatment.•We immediately lyophilized one sample to serve as the control.•We then exposed seven samples outside to natural environmental conditions for 0, 12, 24, 36, 48, 60, or 72 h. After outdoor exposure, we desiccated a subsample in silica beads for an additional 5, 7, or 10 days.•We analyzed the fecal hormone metabolite concentrations in each sample using enzyme immunoassays for corticosterone, cortisol, testosterone, progesterone, and estradiol.We determined that male and female fecal hormone metabolites in fresh African lion fecal samples are stable and comparable to a standardized desiccation method if dried in silica beads for 5 to 10 days prior to storing them at -20℃.


Understanding the contextual functions of C1q and LAIR-1 and their applications.
The importance of the complement component C1q has been highlighted by its involvement in autoimmunity, infection, inflammatory diseases, and tumors. The unique tulip-like structure of C1q has both a collagen-like stalk (C1q tail) and heterotrimeric globular head (gC1q), each with different binding specificities, and the binding of these components to their respective receptors leads to functional complexities in the body and bridges innate and adaptive immunity. This review describes the fundamental roles of C1q in various microenvironments and focuses on the importance of the interactions of C1q and its receptors with the inhibitory receptor LAIR-1 in maintaining homeostasis. Current therapeutic opportunities modulating LAIR-1 are also discussed.


Serologic responses of infertile women to the 60-kd chlamydial heat shock protein (hsp60).
To determine whether women with Chlamydia trachomatis-associated tubal infertility are more likely than other infertile women to have antibodies to a particular region of the 60-kd chlamydial heat shock protein, hsp60. Serologic responses to the chlamydial hsp60 were examined in 43 infertile women seropositive for Chlamydia trachomatis, including 21 women with tubal infertility, 13 women with endometriosis, and 9 women with other causes of infertility. Antibody responses were localized to regions of hsp60 using five nonoverlapping recombinant polypeptides. Sixteen women with tubal infertility had anti-hsp60 antibodies compared with seven women with endometriosis and two women with other causes of infertility. Antibodies of 11 women with tubal infertility reacted predominantly with a region of hsp60 containing amino acids (201 to 300) compared with 1 women without tubal infertility. In contrast, antibodies that localized to the carboxyl terminus, amino acids (401 to 544), were seen equally in all groups. Among seropositive infertile women, antibodies that localized to amino acids (201 to 300) were immunodominant in those with tubal infertility but not in those with infertility due to other causes.


Treatment with the specific estrogen receptor antagonist ICI 182,780 demasculinizes neuron soma size in the developing zebra finch brain.
In zebra finches, many features of the neural song system are more pronounced in males compared to females. The exact mechanism(s) responsible for these differences are unknown, but may involve steroid hormones. More specifically, estrogens are most effective in masculinizing the female brain. Attempts to prevent masculine development through various estrogen receptor antagonists have been relatively ineffective, possibly due to partial agonistic activity of the compounds tested. To further investigate the role of estrogens in dimorphic development we utilized a more potent estrogen receptor blocker, ICI 182,780. Animals were treated during the first 25 days post-hatching. Daily intracranial injections significantly decreased neuron soma size in RA and HVC of both sexes. A similar effect was noted in LMAN. Treatment also appeared to decrease the volume of several song control nuclei. Together, these data support the hypothesis that ICI is an effective estrogen receptor antagonist in the zebra finch brain and that estrogens may influence sexually dimorphic development of the zebra finch song circuit. However, reported inconsistencies about sex differences in estrogen exposure and/or utilization exist, suggesting that complete sexual differentiation and development likely involve additional factors.


Cytotoxic T Lymphocyte Antigen 4 Gene +49 A/G (rs231775) Polymorphism and Susceptibility to Systemic Lupus Erythematosus.
To assess the probable role of +49AG polymorphism in susceptibility to SLE in an Egyptian population. Systemic lupus erythematosus (SLE) is a compound inflammatory chronic disease distinguished through the release of autoantibodies. Cytotoxic T lymphocyte associated antigen-4 is a main down controller of T-cell response; its dysregulation could affect SLE pathogenesis by altered T cells activation to self-antigens. To evaluate the CTLA-4 +49AG allelic and genotype frequency in a sample of the Egyptian population and correlate them with disease susceptibility and clinical severity. Including 100 patients with SLE and 100 healthy controls (age and gender matched), CTLA-4 exon 1 49 A>G Genotyping was done using Real-Time PCR. No difference was noticed in genotype or allele distributions of the studied polymorphism between both groups. Similar genotypes and allele frequencies were established for the 2 groups after their stratification by the age of disease onset, clinical course, or severity. CTLA-4 +49AG gene polymorphism is not linked with the liability to develop SLE in the studied Egyptian population. Yet it is significantly related to disease severity.


Differential expression of cystathionine beta synthase in adolescents with Down syndrome: impact on adiposity.
Obesity is more prevalent among people with Down Syndrome (DS) compared to general population. In this pilot study, we investigated the effect of cystathionine beta-synthase (CBS) overdosage on the regulation of transsulfuration pathway and the obesity phenotype in fifty adolescents (25 obese/overweight and 25 lean) with trisomy 21. The transcriptional levels of CBS in leukocytes and its translational levels in plasma were quantified using real time polymerase chain reaction and enzyme-linked immunosorbent assay respectively. Meanwhile, ultra performance liquid chromatography tandem mass spectrometry was used to determine the plasma concentrations of methionine, homocysteine, cystathionine and cysteine. Fasting plasma lipid profiles were assessed by colorimetric assays. The anthropometric measurements and indices of all subjects were recorded. Both DS groups had comparable levels of CBS transcripts (p = 0.2734). The plasma levels of the enzyme were significantly higher in the lean DS cases (p = 0.0174) compared to the obese/overweight participants. Total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein, methionine, homocysteine, cystathionine and cysteine showed similar plasma levels in both groups. However, the plasma cysteine levels exceeded the normal range in all DS cases. We reported a statistically significant inverse association between CBS enzyme levels and weight (r= - 0.3498, p = 0.0128), hip circumference (r= - 0.3584, p = 0.0106), body mass index (r= - 0.3719, p = 0.0078) and body adiposity index (r= - 0.3183, p = 0.0243). Our data suggests that the high concentrations of CBS enzyme together with cysteine modulate the DS obesity presumably through increased hydrogen sulfide production which has recently showed anti-adiposity effects.


The cardiovascular effects of centrally administered substance P in the anaesthetised rabbit.
The cardiovascular effects of Substance P (SP) administered into the cisterna magna (i.c.) of the pentobarbitone anaesthetised rabbit have been investigated. SP given i.c. in low doses (0.2-10 ng/kg) caused a dose related rise in blood pressure and bradycardia. Both responses were attenuated by pretreatment with intravenous (i.v.) pentolinium. SP given i.v. (1 ng/kg-1 microgram/kg) caused a dose related fall in pressure and tachycardia. The pressor response observed after i.c. administration was reduced by pretreatment with morphine and enhanced by naloxone. Bilateral sinoaortic denervation also enhanced the pressor effect of SP, but after deafferentation naloxone had no further effect. Pretreatment with clonidine abolished this rise in pressure. It is concluded that the central administration of SP has a direct pressor effect whereas the bradycardia appears to result from both direct and indirect effects, leading to a decrease in cardiac sympathetic activity. Both actions are mediated by the autonomic nervous system. SP may be involved in the central regulation of blood pressure and modulation of baroreceptor reflex activity.


Anti-ganglioside antibodies alter presynaptic release and calcium influx.
Acute motor axonal neuropathy (AMAN) variant of Guillain-Barré syndrome is often associated with IgG anti-GM1 and -GD1a antibodies. The pathophysiological basis of antibody-mediated selective motor nerve dysfunction remains unclear. We investigated the effects of IgG anti-GM1 and -GD1a monoclonal antibodies (mAbs) on neuromuscular transmission and calcium influx in hemidiaphragm preparations and in cultured neurons, respectively, to elucidate mechanisms of Ab-mediated muscle weakness. Anti-GM1 and -GD1a mAbs depressed evoked quantal release to a significant yet different extent, without affecting postsynaptic currents. At equivalent concentrations, anti-GD1b, -GT1b, or sham mAbs did not affect neuromuscular transmission. At fourfold higher concentration, an anti-GD1b mAb (specificity described in immune sensory neuropathies) induced completely reversible blockade. In neuronal cultures, anti-GM1 and -GD1a mAbs significantly reduced depolarization-induced calcium influx. In conclusion, different anti-ganglioside mAbs induce distinct effects on presynaptic transmitter release by reducing calcium influx, suggesting that this is one mechanism of antibody-mediated muscle weakness in AMAN.


A novel human RasGAP-like gene that maps within the prostate cancer susceptibility locus at chromosome 1q25.
We report the molecular cloning of a human cDNA that encodes a molecule having striking homology with Ras-specific GTPase-activating proteins (RasGAPs). Among previously described RasGAPs, the cDNA product is most closely related to Caenorhabditis elegans GAP-2, including a predicted coiled-coil structure near the carboxyl terminus. Expression of the cDNA in Saccharomyces cerevisiae defective in one of two RasGAPs, Ira2, complemented loss of the Ira2 function, indicating that the cDNA product functions as a RasGAP. The RasGAP-like gene is located on the human chromosome 1q25, the locus that appears to contain a hereditary prostate cancer susceptible gene, HPC1.


Texture analysis of the liver at MDCT for assessing hepatic fibrosis.
To evaluate CT texture analysis (CTTA) for staging of hepatic fibrosis (stages F0-F4) METHODS: Quantitative texture analysis (QTA) of the liver was performed on abdominal MDCT scans using commercially available software (TexRAD), which uses a filtration-histogram statistic-based technique. Single-slice ROI measurements of the total liver, Couinaud segments IV-VIII, and segments I-III were obtained. CTTA parameters were correlated against fibrosis stage (F0-F4), with biopsy performed within one year for all cases with intermediate fibrosis (F1-F3). The study cohort consisted of 289 adults (158M/131W; mean age, 51 years), including healthy controls (F0, n = 77), and patients with increasing stages of fibrosis (F1, n = 42; F2 n = 37; F3 n = 53; F4 n = 80). Mean gray-level intensity increased with fibrosis stage, demonstrating an ROC AUC of 0.78 at medium filtration for F0 vs F1-4, with sensitivity and specificity of 74% and 74% at cutoff 0.18. For significant fibrosis (≥F2), mean showed AUCs ranging from 0.71-0.73 across medium- and coarse- filtered textures with sensitivity and specificity of 71% and 68% at cutoff of 0.3, with similar performance also observed for advanced fibrosis (≥F3). Entropy showed a similar trend. Conversely, kurtosis and skewness decreased with increasing fibrosis, particularly in cirrhotic patients. For cirrhosis (≥F4), kurtosis and skewness showed AUCs of 0.86 and 0.87, respectively, at coarse-filtered scale, with skewness showing a sensitivity and specificity of 84% and 75% at cutoff of 1.3. CTTA may be helpful in detecting the presence of hepatic fibrosis and discriminating between stages of fibrosis, particularly at advanced levels.


Lipid adaptation in liver mitochondrial membranes of carp acclimated to different environmental temperatures: phospholipid composition, fatty acid pattern and cholesterol content.
The lipid fraction of liver mitochondria has been studied in carp acclimated to high and low environmental temperatures. Evidence is provided for a temperature-induced lipid adaptation which might control membrane fluidity. This supports suggestions made in a recent communication on temperature-induced changes in the Arrhenius functions of mitochondrial oxidase systems from carp liver ((Wodtke, E. (1976) J. Comp. Physiol. 110, 145--157). The results of the analysis of lipid composition are: the ratio of phospholipid to protein does not differ at cold and warm acclimation temperatures. Fish kept at low environmental temperature show a decreased molar ratio of cholesterol : phospholipid in mitochondrial membrane lipids; the diminished complexation renders phospholipids more sensitive to fluidity control by fatty acid substitution. A decrease of mitochondrial phosphatidylcholine at low acclimation temperature is observed, which is compensated by increased amounts of phosphatidylethanolamine and phosphatidylinositol. This means there is an increase in the acidic character of the phospholipids at low environmental temperature, and might be the cause of fluidization of the membrane and a decrease in transition temperature. The fatty acid pattern of carp mitochondria differs markedly from that of mammalian mitochondria; it is not identical for total lipids, phosphatidylcholine and phosphatidylethanolamine, and is roughly characterized by high amounts in n--3, but low amounts in n--6 and mono-unsaturated fatty acids. Low environmental temperature decreases the proportion of saturated species and markedly lowers the ratio of the alpha-linolenic acid family, the latter perhaps being caused by the increased activity of delta6-desaturase, as calculated for low acclimation temperature. An increase in mean unsaturation and in the proportion of 1,2-diunsaturated phospholipids most probably increases membrane fluidity at low acclimation temperature.


How Can the Tuberculosis Laboratory Aid in the Patient-Centered Diagnosis and Management of Tuberculosis?
In 2019, tuberculosis is still a global source of morbidity and mortality. To determine and provide the most effective treatment regimen to patients, the tuberculosis laboratory needs to rapidly but reliably answer 2 main questions: (1) Is Mycobacterium tuberculosis detectable in the patient specimen? and (2) If so, is the strain detected drug susceptible or does it show any form of drug resistance? In cases of drug resistance, health care providers need to have access to minimal inhibitory concentration results and to the type of mutation conferring drug resistance to tailor the most appropriate drug regimen.


Long-term T-cell-mediated immunity to Epstein-Barr virus in man. III. Activation of cytotoxic T cells in virus-infected leukocyte cultures.
Experiments have been conducted to determine the role played by immune T cells in the regression of EB-virus-induced transformation which is exclusively seen in leukocyte cultures from sero-positive donors. Kinetic studies suggest that, in virus-infected cultures from such donors, a population of T cells proliferates within the first 2 weeks apparently in response to the appearance of virus-infected B cells. This proliferation continues to some extent during the period of regression. Nonspecific induction of T-cell proliferation by PHA did not induce regression in virus-infected cultures from seronegative donors and acutally prevented the regression in seropositive donor cultures. T cells harvested from seropositive donor cultures 11-14 days post infection were generally much more inhibitory to the growth of the autologous EB-virus-transformed cell line than were T cells either freshly prepared from whole blood or harvested from corresponding uninfected cultures; this inhibitory activity was either absent or much diminished when assayed against allogeneic target cell lines. The results suggest that virus-specific memory T cells capable of mounting a cytotoxic response when properly challenged in vitro.


Dideoxynucleoside triphosphates inhibit a late stage of SV40 DNA replication in vitro.
The role of DNA polymerases in the replication of SV40 DNA was studied using a T-antigen-dependent assay supplemented with a human KB cell extract. Inhibition of DNA polymerase alpha by addition of aphidicolin or monoclonal antibodies prevented DNA synthesis, confirming the requirement for this enzyme in replication. The replication process was unaffected by ddTTP at a concentration (5 microM) inhibitory to DNA polymerases beta and gamma, however, higher concentrations of ddTTP (200 microM) caused an apparent accumulation of relaxed circular plasmid with a concomitant decrease in DNA synthesis. An analysis of this replication intermediate indicated that it was formed during the replication reaction and that the replicative cycle was nearly complete. A kinetic study of ddTTP inhibition strongly suggested DNA polymerase epsilon (PCNA-independent DNA polymerase delta) was the target of the inhibitor and that this enzyme functions during the final stages of DNA replication.


The distribution and mechanism of iodotyrosine deiodinase defied expectations.
Iodotyrosine deiodinase (IYD) is unusual for its reliance on flavin to promote reductive dehalogenation under aerobic conditions. As implied by the name, this enzyme was first discovered to catalyze iodide elimination from iodotyrosine for recycling iodide during synthesis of tetra- and triiodothyronine collectively known as thyroid hormone. However, IYD likely supports many more functions and has been shown to debrominate and dechlorinate bromo- and chlorotyrosines. A specificity for halotyrosines versus halophenols is well preserved from humans to bacteria. In all examples to date, the substrate zwitterion establishes polar contacts with both the protein and the isoalloxazine ring of flavin. Mechanistic data suggest dehalogenation is catalyzed by sequential one electron transfer steps from reduced flavin to substrate despite the initial expectations for a single two electron transfer mechanism. A purported flavin semiquinone intermediate is stabilized by hydrogen bonding between its N5 position and the side chain of a Thr. Mutation of this residue to Ala suppresses dehalogenation and enhances a nitroreductase activity that is reminiscent of other enzymes within the same structural superfamily.


Membrane damage mechanism of protocatechualdehyde against Micrococcus luteus and its effect on pork quality characteristics.
This study investigated the mechanism of membrane damage by protocatechualdehyde (PCA) against Micrococcus luteus and assessed effects of PCA on the sensory and physicochemical properties of pork. The mechanism of PCA inhibition on M. luteus was studied by determining the minimum inhibitory concentration (MIC) based on membrane potential, intracellular ATP concentration, intracellular pH, confocal laser scanning microscopy (CLSM), and field emission gun scanning electron microscopy (FEG-SEM). The results showed that the MIC of PCA against M. luteus was 1.25 mg/mL. Hyperpolarization of the bacterial cell membrane, a decrease in the intracellular ATP concentration, and intracellular pH indicated that PCA damaged the cell membrane of M. luteus. FEG-SEM observation revealed that PCA could cause surface collapse, cell membrane rupture, and content outflow of M. luteus. Additionally, PCA was found to inhibit increases in the total number of colonies, the thiobarbituric acid reactive substances (TBARS) value growth rate, and moisture mobility in raw pork. Additionally, it improved the color and texture of raw pork, all of which effectively prolonged its shelf life. This study will encourage the application of PCA as a natural antibacterial agent in the food industry.


Diagnostic NGS for Severe Neuromuscular Disorders.
Investigators from the University of Western Australia report the diagnostic yield of performing next generation sequencing (NGS; whole exome and targeted capture of 277 neuromuscular genes) in a heterogenous cohort of patients with neuromuscular disorders (NMD) presenting at or before birth.


Perceptual organization of tone sequences in the auditory cortex of awake macaques.
Acoustic sequences such as speech and music are generally perceived as coherent auditory "streams," which can be individually attended to and followed over time. Although the psychophysical stimulus parameters governing this "auditory streaming" are well established, the brain mechanisms underlying the formation of auditory streams remain largely unknown. In particular, an essential feature of the phenomenon, which corresponds to the fact that the segregation of sounds into streams typically takes several seconds to build up, remains unexplained. Here, we show that this and other major features of auditory-stream formation measured in humans using alternating-tone sequences can be quantitatively accounted for based on single-unit responses recorded in the primary auditory cortex (A1) of awake rhesus monkeys listening to the same sound sequences.


High-fat diet induces cardiomyocyte apoptosis via the inhibition of autophagy.
Excessive fat intake induces obesity and causes cardiac injury. Intracellular degradation process involving destruction of long-lived proteins and organelles maintains homeostasis for cells under stress. The purpose of this study was to explore the relation of high-fat diet (HFD)-induced cardiac injury and intracellular degradation process with regard to autophagy and ER stress. HFD feeding for 24 weeks induced hyperglycemia, hyperlipidemia, and cardiac hypertrophy in adult male C57BL/6 mice. In the heart, PARP cleavage, an indicator of apoptosis, levels of LC3-II and p62, indicators of autophagy, and CHOP, indicator of ER stress, were increased. A palmitate-treated cardiomyoblast (H9C2) cell culture was examined to explore how HFD induced myocardial injury. Excessive palmitate (400 μM) treatment induced apoptosis and increased the number of autophagosomes and acid vacuoles of H9C2 cells. Besides, it elevated the expression of LC3-II, p62, and PARP cleavage. Induction of autophagy by rapamycin ameliorated palmitate-induced apoptosis, while inhibition of autophagy by 3-methyladenine or LC3 siRNA exacerbated palmitate-induced apoptosis. Palmitate treatment also induced CHOP expression which is associated with ER stress. HFD can cause cardiac injury by induction of apoptosis which is associated with autophagy dysregulation and ER stress. In addition, autophagy deficiency augments cardiac apoptosis, suggesting that autophagy serves as a pro-survival role in lipotoxic condition.


Potential role of human umbilical cord stem cells-derived exosomes as novel molecular inhibitors of hepatocellular carcinoma growth.
Hepatocellular carcinoma (HCC) is one of the most critical cancers; thus, novel therapeutical regimens are of great need. In this study, we investigated the effects of umbilical cord mesenchymal stem cells (UC-MSCs) derived exosomes on HepG2 cell line, and the underlying mechanism to control HCC proliferation, to identify the potential clinical role of exosomes as a novel molecular therapeutic target. Proliferation, apoptosis, and angiogenesis effects were assessed together with the cell viability evaluation by MTT assay in HepG2 cells at 24/48 h. with or without UC-MSCs-derived exosomes. Gene expressions of TNF-α, caspase-3, VEGF, stromal cell-derived factor-1 (SDF-1), and CX chemokine receptor-4 (CXCR-4) were measured by quantitative real-time PCR technique. Expression of sirtuin-1 (SIRT-1) protein was detected by western blot. Treatment of HepG2 cells with UC-MSCs-derived exosomes for 24 and 48 h. demonstrated a significant reduction of cells survival compared to the control group (p < 0.05). The SIRT-1 protein, and VEGF, SDF-1, CXCR-4 expression levels were significantly lower, TNF-α and caspase-3 expression levels were significantly higher in exosomal-treated HepG2 cells for 24 and 48 h. than those in the control group. Moreover, our findings documented that the anti-proliferative, apoptotic, and anti-angiogenic effects were achieved in a time-dependent manner in which more effects were determined after 48 h supplementation compared to 24 h (p < 0.05). UC-MSCs-derived exosomes exert anticarcinogenic molecular effects on HepG2 cells through the involvement of SIRT-1, SDF-1, and CXCR-4. Hence, exosomes would be a potential novel therapy regimen against HCC. Large-scale studies are recommended to verify this conclusion.


Clinical and radiological differences in posterior reversible encephalopathy syndrome between patients with preeclampsia-eclampsia and other predisposing diseases.
Posterior reversible encephalopathy syndrome (PRES) is a serious maternal complication in pregnancy, but data on the clinicoradiological differences to other etiologies of PRES are scarce. In this study, we aimed to investigate the clinical and imaging characteristics of PRES in preeclampsia-eclampsia patients compared with other predisposing diseases in a large cohort. We retrospectively reviewed the radiological report data bases between January 1999 and August 2010 for patients with PRES. Patients fulfilling the criteria for PRES after detailed investigation of clinical charts and imaging studies were separated into patients with eclampsia-preeclampsia and other predisposing causes. Various imaging features at onset of symptoms and on follow-up as well as clinical and paraclinical data were analyzed. A total of 24 patients with preeclampsia-eclampsia associated PRES and 72 patients with PRES of other predisposing causes were included into the study. In preeclampsia-eclampsia patients, headaches were significantly more frequent as initial PRES-related symptom (P < 0.001), whereas altered mental state was significantly less frequent (P = 0.001). Thalamus, midbrain, and pons affection was significantly less frequent in preeclampsia-eclampsia associated PRES (P = 0.01). Preeclampsia-eclampsia patients had significantly less severe edema, less cytotoxic edema, hemorrhage and contrast enhancement, while more frequent complete resolution of edema and less frequent residual structural lesions were seen on follow-up imaging. In our PRES cohort, we found major clinicoradiological differences between preeclampsia-eclampsia and other predisposing causes pointing toward a less severe course of disease in preeclampsia-eclampsia.


Anti-Tumor Effects of Second Generation β-Hydroxylase Inhibitors on Cholangiocarcinoma Development and Progression.
Cholangiocarcinoma (CCA) has a poor prognosis due to widespread intrahepatic spread. Aspartate β-hydroxylase (ASPH) is a transmembrane protein and catalyzes the hydroxylation of aspartyl and asparaginyl residues in calcium binding epidermal growth factor (cbEGF)-like domains of various proteins, including Notch receptors and ligands. ASPH is highly overexpressed (>95%) in human CCA tumors. We explored the molecular mechanisms by which ASPH mediated the CCA malignant phenotype and evaluated the potential of ASPH as a therapeutic target for CCA. The importance of expression and enzymatic activity of ASPH for CCA growth and progression was examined using shRNA "knockdown" and a mutant construct that reduced its catalytic activity. Second generation small molecule inhibitors (SMIs) of β-hydroxylase activity were developed and used to target ASPH in vitro and in vivo. Subcutaneous and intrahepatic xenograft rodent models were employed to determine anti-tumor effects on CCA growth and development. It was found that the enzymatic activity of ASPH was critical for mediating CCA progression, as well as inhibiting apoptosis. Mechanistically, ASPH overexpression promoted Notch activation and modulated CCA progression through a Notch1-dependent cyclin D1 pathway. Targeting ASPH with shRNAs or a SMI significantly suppressed CCA growth in vivo.


Influence of extract purification on the recovery of citrinin in the HPLC analysis.
Various analytical methods of citrinin determination are known from publications which cannot be applied without any difficulties. Low fluctuating recovery rates of standard additions, an insufficient extract purification for rye, and a rather low sensitivity based on improper HPLC conditions are being observed if the published methods are applied. An easily applicable, reliable and sensitive HPLC method in the lower ppb range on the basis of a solid phase extraction combined with a HPLC gradient which allows for a sensitive fluorescence detection, is under development. The results already achieved are described in the following.


Stimulation of synaptosomal Na+,K(+)-ATPase by ethanol: possible involvement of an isozyme-specific inhibitor of Na+,K(+)-ATPase.
In synaptosomal preparations from rat cerebral cortex, ouabain-sensitive Rb+ uptake was stimulated by ethanol (20-80 mM). Based on differential sensitivity to ouabain, 80% of this Na+,K(+)-ATPase activity represented activity of the alpha 1 isozyme while 20% was due to the alpha 2 and/or alpha 3 isozymes (alpha 2/ alpha 3). Stimulation of Na+,K(+)-ATPase was selective for the activity of alpha 2/alpha 3 which was increased by 167% in the presence of 80 mM ethanol. In this concentration range, ethanol had no effect on alpha 1 activity. Exposure of synaptosomal preparations to EGTA increased basal (no ethanol) alpha 2/alpha 3 activity with no effect on alpha 1 activity. Further, ethanol no longer stimulated alpha 2/alpha 3 activity after EGTA treatment. An EGTA extract was concentrated and desalted to yield a fraction that selectively inhibited alpha 2/alpha 3 activity when reconstituted with EGTA-treated synaptosomal preparations. This inhibition was trypsin-sensitive, suggesting protein involvement, and was prevented by 80 mM ethanol. In the presence of the inhibitory protein fraction, ethanol stimulated Na+, K(+)-ATPase activity in EGTA-treated membranes with a dose-response like that observed with the crude (no EGTA) synaptosomes. We propose that the alpha 2/alpha 3 activity of Na+,K(+)-ATPase is subject to inhibitory regulation and that ethanol stimulates this activity by releasing it from inhibition, an effect that may mimic in vivo deregulation of the enzyme by ethanol.


Optimal indications of endocrine therapy alone as adjuvant systemic treatment of breast cancer.
We examined the validity of the St Gallen algorithm for Japanese breast cancer patients and sought the optimal indications of endocrine monotherapy as adjuvant systemic treatment. According to the 2005 St Gallen algorithm, endocrine responsiveness (responsive, uncertain, or non-responsive) and recurrence risk (low, intermediate, or high) were assessed in 436 invasive breast cancer patients, who underwent surgery and adjuvant therapy of tamoxifen alone in 1982-1993. Furthermore, intermediate-risk patients were divided into three groups based on lymph node metastasis and number of risk factors as follows: Group A, negative lymph node metastasis and one risk factor; Group B, negative lymph node metastasis and two to five risk factors; and Group C, positive lymph node metastasis. Cumulative 10-year recurrence-free survival (RFS) rates of each type were calculated. Recurrence-free survival was as follows: endocrine responsiveness; responsive: 86.0%, uncertain: 79.5%, non-responsive: 72.4%, risk category; low: 93.3%, intermediate: 84.0%, high: 59.6%, intermediate-risk patients; Group A: 93.5%, Group B: 88.2%, and Group C: 75.0%. In conclusion, patient classification based on St Gallen algorithm appears valid in Japanese breast cancer patients. Endocrine monotherapy may be sufficient as adjuvant treatment in the intermediate-risk patients, in which only one risk factor was present without any metastatic involvement in lymph node.


Evaluating deep-sea communities' susceptibility to mining plumes using shallow-water data.
Increased suspended sediment concentrations (SSC) are a major stressor across aquatic habitats. Here, the literature was synthesized to show that animal responses to increases in relative SSC (test concentration/natural background concentration) were similar in type and negative across different shallow-water (marine, estuarine, freshwater) habitats. Further, animal sensitivities are similar across habitats based on relative SSC and occur starting at low relative SSC increases in all habitats despite differences in natural background SSC. Based upon these similarities in relative SSC sensitivities, deep-sea sensitivity values for acute exposure to increased SSC, where empirical data are almost non-existent, were estimated. Because of the low natural SSC in deep sea environments, very small increases in absolute SSC could result in acute effects. How the methods and results can be used to inform regulatory thresholds are discussed. Because of the large variability in shallow water datasets and differences between deep-sea and shallow-water habitats, deep-sea specific data are needed to verify the estimates and improve their precision. Following the precautionary principle and the results presented here, it is recommended that the threshold for acute plume impacts is set very close to natural background levels.


No post-genetics era in human disease research.
In the 1980s, linkage emerged as a route to discovering genetic defects, spurring the rise of genomics and making gene-based approaches available to previously phenotype-orientated researchers. In the post-genomics era, genetics is fundamental to understanding disease at all stages of the pathogenic process.


Soft tissue non-Hodgkin lymphoma of shoulder in a HIV patient: a report of a case and review of the literature.
The risk of developing lymphoma is greatly increased in HIV infection. Musculoskeletal manifestations of the human immunodeficiency virus (HIV) are common and are sometimes the initial presentation of the disease. Muscle, bone, and joints are involved by septic arthritis, myopathies and neoplasms. HIV-related neoplastic processes that affect the musculoskeletal system include Kaposi's sarcoma and non-Hodgkin's lymphoma, the latter being mainly localized at lower extremities, spine and skull. The Authors report a case of a 34 year-old lady. In December 2003 the patient noted a painless mass on her right shoulder whose size increased progressively. In March 2004 she was diagnosed HIV positive and contemporary got pregnant. The patient decided to continue her pregnancy and to not undergo any diagnostic procedure and treatment. At the end of August she underwent a surgical ablation of the lesion that revealed a lesion of 7 cm x 7 cm x 3.3 cm. The histology showed B-cells expressing CD20, PAX-5, CD10, BCL-6 and MUM-1 with 70% Ki67 positive nuclei. The lesion was also negative for EBV infection and showed a monoclonal rearrangement of IgH chain and a polyclonal pattern for TCR gamma and beta. A final diagnosis of diffuse large B-cell lymphoma was made. The patient underwent postoperative chemotherapy. At four-years follow up the patient is symptom free and no local nor systemic recurrence of pathology has been noted on MRI control. HIV infection is still under control. In this report, we present a case of diffuse large B-cell lymphoma localized in the soft tissue of the shoulder in a HIV infected patient. Authors want to underline this case for the rare position, the big size and the association with HIV infection.


Correlations between aqueous flare and cells and lens surface cytology in eyes with poly(methyl methacrylate) and heparin-surface-modified intraocular lenses.
A prospective, single-surgeon study was performed on 59 patients randomized to receive a poly(methyl methacrylate) intraocular lens or a heparin-surface-modified lens after standardized endocapsular cataract surgery with a routine postoperative regimen. The anterior chamber laser flare and cell measurements and the cellular reaction on the anterior lens surface were monitored for a year following surgery. This report examines the correlations between the anterior chamber flare and cell reaction and the anterior IOL surface cellular reaction. The findings suggest that the early postoperative blood-aqueous barrier breakdown is primarily caused by surgical trauma; by one month the blood-aqueous barrier function is determined by inflammatory mediators released by small monocyte-derived cells on the IOL surface. Giant cells on the IOL surface did not correlate with aqueous flare, which suggests an insignificant capacity to release inflammatory mediators.


Zinc Acts Synergistically with Berberine for Enhancing Its Efficacy as an Anti-cancer Agent by Inducing Clusterin-Dependent Apoptosis in HT-29 Colorectal Cancer Cells.
It is now widely accepted that anti-cancer medications are most effective when administered in combination. Zinc is an essential micronutrient whilst berberine is a well-known natural phytochemical, both having multiple molecular mechanisms of action. The present study aimed to determine the combinatorial effect of zinc and berberine on the human adenocarcinoma HT-29 cancer cell line. The anti-proliferative activity of berberine and zinc was determined by cell viability and colony-forming assays. The combination index and drug reduction index values of zinc and berberine co-treatments were estimated by suitable software. Flow cytometry was used to analyse cell cycle distribution and Annexin V/PI staining. The expression of apoptosis and zinc signalling markers were analysed by RT-qPCR and immunoblot analysis. Berberine decreased the viability of colon cancer cells in a dose-dependent manner whilst zinc alone had no significant influence on it. However, zinc and berberine co-treatment resulted in a synergistic anti-cancer action which was demonstrated by G2/M phase arrest of cell growth at a lower dose of berberine. Annexin V assay demonstrated that the synergistic impact of zinc and berberine boosted the number of apoptotic cells. Gene expression analysis at both transcriptional and translational levels showed the upregulation of apoptotic (caspase-3 and caspase-8) and a zinc-sensing receptor (GPR39) gene with concomitant downregulation of anti-apoptotic genes like proliferating cell nuclear antigen (PCNA) and clusterin. Our findings showed that the combination of zinc and berberine has synergistic anti-cancer efficacy and thus could be used as a potential chemopreventive option for colon cancer.


Cholesterol depletion inhibits src family kinase-dependent calcium mobilization and apoptosis induced by rituximab crosslinking.
The monoclonal antibody (mAb) rituximab produces objective clinical responses in patients with B-cell non-Hodgkin's lymphoma and antibody-based autoimmune diseases. Mechanisms mediating B-cell depletion by rituximab are not completely understood and may include direct effects of signalling via the target antigen CD20. Like most but not all CD20 mAbs, rituximab induces a sharp change in the solubility of the CD20 protein in the non-ionic detergent Triton-X-100, reflecting a dramatic increase in the innate affinity of CD20 for membrane raft signalling domains. Apoptosis induced by rituximab hypercrosslinking has been shown to require src family kinases (SFK), which are enriched in rafts. In this report we provide experimental evidence that SFK-dependent apoptotic signals induced by rituximab are raft dependent. Cholesterol depletion prevented the association of hypercrosslinked CD20 with detergent-insoluble rafts, and attenuated both calcium mobilization and apoptosis induced with rituximab. CD20 cocapped with the raft-associated transmembrane adaptor LAB/NTAL after hypercrosslinking with CD20 mAbs, regardless of their ability to induce a change in the affinity of CD20 for rafts. Taken together, the data demonstrate that CD20 hypercrosslinking via rituximab activates SFKs and downstream signalling events by clustering membrane rafts in which antibody-bound CD20 is localized in a high-affinity configuration.


Predicting Neurocognitive Change after Bilateral Deep Brain Stimulation of Subthalamic Nucleus for Parkinson's Disease.
Deep brain stimulation (DBS) of the bilateral subthalamic nucleus (STN) is a standard surgical treatment option in patients with advanced Parkinson's disease. Adverse effects on cognitive function have been reported, impacting the quality of life of patients and caregivers. We aimed to investigate a quantitative predictive preexisting cognitive factor for predicting postoperative cognitive changes. Thirty-five patients underwent STN-DBS. A battery of neuropsychological tests were used to examine executive function, processing speed, and visuospatial function both preoperatively and 1 year postoperatively. A multiple logistic regression analysis was performed to investigate the relationships between preoperative factors and cognitive outcomes. The predictive value of the preoperative factors for global cognitive decline during long-term follow-up were evaluated. The patients exhibited significant changes in processing speed and visuospatial function after surgery. Using reliable change index values, lower preoperative scores on the Similarities and Object Assembly subtests of the Wechsler Adult Intelligence Scale III were associated with decreases in visuospatial function at 1 year after DBS. The odds ratios were 10.2 for Similarities and 9.53 for Object Assembly. The proportion of Mini Mental State Examination-maintained patients with low scores on the Similarities subtest was significantly lower than that of patients with high scores at 3 and 5 years. No factors were found to be related to decreases in processing speed. Preoperative evaluation of the Similarities and Object Assembly subtests may be useful to identify patients who are at a greater risk of experiencing decreases in visuospatial functioning after STN-DBS. Furthermore, a low score on the Similarities subtest may predict future global cognitive deterioration.


Review of doped silica glass optical fibre: their TL properties and potential applications in radiation therapy dosimetry.
Review is made of dosimetric studies of Ge-doped SiO(2) telecommunication fibre as a 1-D thermoluminescence (TL) system for therapeutic applications. To-date, the response of these fibres has been investigated for UV sources, superficial X-ray beam therapy facilities, a synchrotron microbeam facility, electron linear accelerators, protons, neutrons and alpha particles, covering the energy range from a few eV to several MeV. Dosimetric characteristics include, reproducibility, fading, dose response, reciprocity between TL yield and dose-rate and energy dependence. The fibres produce a flat response to fixed photon and electron doses to within better than 3% of the mean TL distribution. Irradiated Ge-doped SiO(2) optical fibres show limited signal fading, with an average loss of TL signal of ~0.4% per day. In terms of dose response, Ge-doped SiO(2) optical fibres have been shown to provide linearity to x and electron doses, from a fraction of 1 Gy up to 2 kGy. The dosimeters have also been used in measuring photoelectron generation from iodinated contrast media; TL yields being some 60% greater in the presence of iodine than in its absence. The review is accompanied by previously unpublished data.


New insights into the mechanochemical coupling mechanism of kinesin-microtubule complexes from their high-resolution structures.
Kinesin motor proteins couple mechanical movements in their motor domain to the binding and hydrolysis of ATP in their nucleotide-binding pocket. Forces produced through this 'mechanochemical' coupling are typically used to mobilize kinesin-mediated transport of cargos along microtubules or microtubule cytoskeleton remodeling. This review discusses the recent high-resolution structures (<4 Å) of kinesins bound to microtubules or tubulin complexes that have resolved outstanding questions about the basis of mechanochemical coupling, and how family-specific modifications of the motor domain can enable its use for motility and/or microtubule depolymerization.


Simple and sensitive determination of 2,4-xylenol in surface water samples from river and sea by gas chromatography-mass spectrometry.
A simple and selective method was developed for determination of the concentration of 2,4-xylenol in river and sea water samples using gas chromatography/mass spectrometry (GC/MS). Trace amounts of 2,4-xylenol were collected in a Oasis HLB Plus cartridge, eluted with acetonitrile. The method detection limit of 2,4-xylenol was 1.4 ng/L. The trace peaks of 2,4-xylenol were found in water samples from the river and the sea, and the concentrations were all less than 1.4 ng/L. The nine peaks of the 2,3-, 2,4-, 2,5-, 2,6-, 3,4-, 3,5-xylenol, and o-, m-, p-ethyl phenol that gave the same m/z ratio were separated efficiently.


Pulmonary extravascular fluid accumulation in recreational climbers: a prospective study.
High altitude pulmonary oedema (HAPE) that is severe enough to require urgent medical care is infrequent. We hypothesised that subclinical HAPE is far more frequent than suspected during even modest climbs of average effort. We assessed 262 consecutive climbers of Monte Rosa (4559 m), before ascent and about 24 h later on the summit 1 h after arriving, by clinical examination, electrocardiography, oximetry, spirometry, carbon monoxide transfer, and closing volume. A chest radiograph was taken at altitude. Only one climber was evacuated for HAPE, but 40 (15%) of 262 climbers had chest rales or interstitial oedema on radiograph after ascent. Of 37 of these climbers, 34 (92%) showed increased closing volume. Of the 197 climbers without oedema, 146 (74%) had an increase in closing volume at altitude. With no change in vital capacity, forced expiratory volume in 1 s and forced expiratory flow at 25-75% of forced vital capacity increased slightly at altitude, without evidence of oedema. If we assume that an increased closing volume at altitude indicates increased pulmonary extravascular fluid, our data suggest that three of every four healthy, recreational climbers have mild subclinical HAPE shortly after a modest climb. The risk of HAPE might not be confined to a small group of genetically susceptible people, but likely exists for most climbers if the rate of ascent and degree of physical effort are great enough, especially if lung size is normal or low.


Role of 4-hydroxynonenal in stress-mediated apoptosis signaling.
In this mini review we summarize recent studies from our laboratory, which show the involvement of 4-hydroxynonenal (4-HNE) in cell cycle signaling. We demonstrate 4-HNE induced apoptosis in various cell lines is accompanied with c-Jun-N-terminal kinase and caspase-3 activation. Cells exposed to mild, transient, heat or oxidative stress acquire capacity to exclude intracellular 4-HNE at a faster rate by inducing hGST5.8 which conjugate 4-HNE to GSH, and RLIP76 which mediates the ATP-dependent transport of the GSH-conjugate of 4-HNE. The cells preconditioned with mild transient stress acquire resistance to H(2)O(2) and 4-HNE induced apoptosis by excluding intracellular 4-HNE at an accelerated pace. Furthermore, a decrease in intracellular concentration of 4-HNE achieved by transfecting cells with mGSTA4-4 or hGSTA4-4 results in a faster growth rate. These studies strongly suggest a role of 4-HNE in stress mediated signaling.


Poststreptococcal dystonia with bilateral striatal enlargement: MR imaging and spectroscopic findings.
Isolated bilateral striatal necrosis is an abnormality of the basal ganglia associated with acute dystonia in children. This report describes the development of dystonic movements in a 7-year-old male patient 2 weeks after streptococcal pharyngitis.


BtuB TonB-dependent transporters and BtuG surface lipoproteins form stable complexes for vitamin B12 uptake in gut Bacteroides.
Vitamin B12 (cobalamin) is required for most human gut microbes, many of which are dependent on scavenging to obtain this vitamin. Since bacterial densities in the gut are extremely high, competition for this keystone micronutrient is severe. Contrasting with Enterobacteria, members of the dominant genus Bacteroides often encode several BtuB vitamin B12 outer membrane transporters together with a conserved array of surface-exposed B12-binding lipoproteins. Here we show that the BtuB transporters from Bacteroides thetaiotaomicron form stable, pedal bin-like complexes with surface-exposed BtuG lipoprotein lids, which bind B12 with high affinities. Closing of the BtuG lid following B12 capture causes destabilisation of the bound B12 by a conserved BtuB extracellular loop, causing translocation of the vitamin to BtuB and subsequent transport. We propose that TonB-dependent, lipoprotein-assisted small molecule uptake is a general feature of Bacteroides spp. that is important for the success of this genus in colonising the human gut.