Unusual complexation behavior between daclatasvir and γ-Cyclodextrin. A multiplatform study.
During a screening of cyclodextrins (CDs) as chiral selectors for the separation of daclatasvir (DCV) and its enantiomer by capillary electrophoresis (CE), an unusual phenomenon for CDs was observed, that is two peaks with a plateau in between using γ-CD as chiral selector. The same result was encountered when enantiopure DCV was injected or when analyzing a sample containing enantiopure DCV and γ-CD in a CD-free background electrolyte. Peak coalescence was observed at 45°C and at a pH above 3.5. Two peaks with a plateau were also observed for DCV stereoisomers as well as a structural analog. However, only a single peak was detected if one or both amino acid moieties of DCV were lacking. Nuclear magnetic resonance (NMR) experiments including Nuclear Overhauser effect-based methods showed that in solution DCV adopted a folded conformation in which the isopropyl side chain of the valine residues pointed toward the aromatic rings of DCV. Moreover, NMR unequivocally demonstrated the simultaneous formation of DCV-γ-CD inclusion complexes with 1:1 and 2:1 stoichiometry, which was corroborated by mass spectrometry. In both complexes, DCV also adopted a folded structure. The RSSR-diastereomer of DCV as well as an analog lacking one of the amino acid moieties also formed 1:1 and 2:1 complexes with γ-CD although a plateau was only observed in the case of the RSSR-diastereomer. As shown by CE-MS, both DCV-γ-CD complexes surprisingly comigrated as the first peak, while the second migrating peak represents non-complexed DCV.


Associated pelvic organ prolapse in women with stress urinary incontinence: when to operate?
This review discusses recently published data concerning the indications for pelvic organ prolapse (POP) surgery in women who present with stress urinary incontinence (SUI). POP can functionally mask SUI. Surgery for POP may unmask occult SUI in many women. Clinically continent women undergoing POP surgery are at risk for developing symptomatic SUI postoperatively. Preoperative identification of occult (and overt) SUI will facilitate the use of an appropriate prophylactic anti-incontinence procedure at the time of prolapse repair. Numerous studies on the preoperative prediction of SUI following repair of POP have been conducted in an effort to determine whether concomitant prophylactic measures should be taken at the time of POP repair to prevent the postoperative unmasking of SUI in women who do not have SUI preoperatively (with or without prolapse reduction). Although the literary evidence available is not sufficient for POP with occult SUI, there is some information available to guide clinicians in deciding when to perform concurrent POP surgery in women who are undergoing primary surgery for SUI. The intended goal of surgical correction of SUI and POP is durable restoration of normal anatomy and function, with symptomatic relief and avoidance of morbidity. Recommendations regarding when to surgically intervene for POP in women who present with SUI are based on the available literature although contemporary studies are few and include small numbers of patients with no controls. Long-term, randomized, controlled prospective studies of large numbers of patients are indicated.


Performance of a PCR assay for the rapid identification of the Klebsiella pneumoniae ST258 epidemic clone in Latin American clinical isolates.
The worldwide dissemination of Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae ST258 demands a rapid PCR-based typing method to detect unique genes of the ST258 clone. This study evaluates a PCR developed by Adler et al. (2014) for the detection of ST258 in K. pneumoniae clinical isolates centered on the identification of the pilv-I and prp genes. We tested 143 clinical isolates from Argentina (n=109), Chile (n=1), Colombia (n=1), Costa Rica (n=2), Ecuador (n=5), El Salvador (n=2), Nicaragua (n=5), Panamá (n=2), Paraguay (n=2), Perú (n=3) and Trinidad and Tobago (n=11) recovered from 2006 to 2015. blaKPC, pilv-l and prp genes were detected by PCR and sequenced by standard procedures. ST258 and non-ST258 were defined by PFGE and/or MLST. Isolates were grouped according to PFGE patterns: 58 were compatible with ST258 (group 1) and 85 with non-ST258 (group 2). MLST study was done on an arbitrary selection of isolates. The pilv-l gene was present only in ST258 isolates, regardless of the presence of the blaKPC gene. Results for the prp gene were variable. Its presence did not define ST258. The pilv-I PCR had a sensitivity and specificity of 100%, respectively, for the detection of ST258 in the isolates under investigation. Given our findings, the pilv-I PCR could replace more time and resource consuming methods, allowing for more rapid detection of the circulating high risk K. pneumoniae clone ST258 in Latin American (LA) countries.


Effect of benserazide on the levels of pineal 5-hydroxytryptamine, melatonin synthesising enzymes and serum melatonin.
The effect of benserazide, an aromatic L-amino acid decarboxylase inhibitor, has been investigated on pineal 5-hydroxytryptamine content, melatonin synthesising enzyme activities and serum melatonin concn. Increasing doses of benserazide caused increasing reductions in the pineal content of 5-hydroxytryptamine. Dark phase serum melatonin concns were also greatly reduced. The drug abolished the diurnal rhythm of hydroxyindole-O-methyltransferase by increasing the light period activities and decreasing the night levels of this enzyme. Pineal N-acetyltransferase was unaffected. It is concluded that benserazide probably inhibits melatonin synthesis by preventing the formation of the substrate, 5-hydroxytryptamine.


A Novel Direct Approach to the Deep Uterine Vein in Laparoscopic Radical Hysterectomy.
To describe a direct approach to the deep uterine vein in laparoscopic radical hysterectomy. Demonstration of the laparoscopic technique with narrated video footage. Securing sufficient radicality is extremely important when performing a radical hysterectomy for cervical cancer, either by laparotomy or by minimally invasive surgery. The nerve-sparing Okabayashi radical hysterectomy (NS-RH) was originally aimed at achieving both radical resection and function preservation [1-3]. A key procedure when performing NS-RH is intraoperative identification of the relationship between the deep uterine vein and pelvic splanchnic nerve fibers [4]. With this in mind, a safe and easy method for identifying the crossing point of the deep uterine vein and pelvic splanchnic nerve in the initial phase of the surgery may greatly improve the safety and efficacy of functional preservation in NS-RH. Herein, we describe a minimally invasive "direct approach" to the deep uterine vein. Before undergoing the pelvic lymphadenectomy, all steps of laparoscopic radical hysterectomy were performed. First, we identified the ureter on the posterior peritoneum, and the peritoneum was dissected just above the ureter. By continuously exploring the pelvic cavity along the ureter, especially through the opening of the space below the ureter in a cranial to caudal direction, we could easily identify the deep uterine vein. This procedure also exposed the fibers of the hypogastric nerve, clarifying the relationship of these structures. Because the relationship between the deep uterine vein and nerve fibers is the most important guidepost of this surgery, their identification in the early phase of the surgery enables us to perform the subsequent procedure precisely and securely. This direct approach to the deep uterine vein can be easily and safely performed.


Nano-vaccines combining customized in situ anti-PD-L1 depot for enhanced tumor immunotherapy.
Low response rate of immune checkpoint blockade (ICB) has limited its clinical application. A promising strategy to overcome this limitation is the use of therapeutic cancer vaccines, which aim to induce robust immune responses that synergize with ICB through immune enhancement and immune normalization strategies. Herein, we developed a combination immunotherapy by combining nano-vaccines consisting of whole tumor cell lysates/CpG liposomes (LCLs) with an anti-PD-L1 loaded lipid gel (aPD-L1@LG). The LCLs were fabricated using cationic liposomes, while the lipid gels (LGs) were prepared by using soybean phosphatidylcholine (SPC) and glycerol dioleate (GDO). Subcutaneous administration of LCLs successfully activated dendritic cells (DCs), and intratumoral administration of anti-PD-L1@LG ensured sustained ICB activity. These results demonstrated that this combination immunotherapy enhanced anti-tumor efficacy and prolonged the survival time in melanoma by activating systemic anti-tumor immune responses. These findings highlight the potential of this rational design as a promising strategy for tumor treatment.


In vitro chemotherapy sensitivity patterns of human colon carcinoma continuous cell lines.
The cells obtained from four established human colon carcinoma cell lines were cultured in soft agar in the presence of eight chemotherapeutic agents that have been used clinically in patients with metastatic colorectal carcinoma. 5-Fluorouracil, mitomycin C, adriamycin, and cis-platinum were highly cytotoxic in vitro at the concentrations employed. These findings were in marked contrast to in vitro chemotherapy sensitivity patterns previously reported by us for a large series of primary human colorectal carcinomas cultured in soft agar. The possible reasons behind these differences are discussed.


Dose escalation phase 1 study of radiotherapy in combination with anti-cytotoxic-T-lymphocyte-associated antigen 4 monoclonal antibody ipilimumab in patients with metastatic melanoma.
A synergy between radiotherapy and anti-cytotoxic-T-lymphocyte-associated antigen 4 (anti-CTLA-4) monoclonal antibody has been demonstrated preclinically. The Mel-Ipi-Rx phase 1 study aimed to determine the maximum tolerated dose (MTD) and safety profile of radiotherapy combined with ipilimumab in patients with metastatic melanoma. A 3+3 dose escalation design was used with 9, 15, 18 and 24 Gy dose of radiotherapy at week 4 combined with 10 mg/kg ipilimumab every 3 weeks for four doses. Patients with evidence of clinical benefit at week 12 were eligible for maintenance with ipilimumab 10 mg/kg every 12 weeks starting at week 24 until severe toxicity or disease progression. The database lock occurred on April 30, 2019. Tumor growth rate of irradiated lesions and non-irradiated lesions were analyzed to assess the systemic immunologic antitumor response. Blood immune monitoring was performed before and during treatment to determine if radiotherapy could modify ipilimumab pharmacodynamics. 19 patients received ipilimumab between August 2011 and July 2015. Nine patients received the four doses of ipilimumab. All patients received the combined radiotherapy. Grade 3 adverse events occurred in nine patients, the most common being colitis and hepatitis. No drug-related death occurred. Dose limiting toxicity occurred in two of six patients in the cohort receiving 15 Gy. The MTD was 9 Gy. Two patients had complete response, three had partial response response and seven had stable disease, giving an objective response rate of 31% and a clinical benefit rate of 75% at week 24. The median duration of follow-up was 5.8 years (Q1=4.5; Q3=6.8). The median overall survival (95% CI) was estimated at 0.9 years (0.5-2). The median progression-free survival (PFS) (95% CI) was 0.4 (0.2-1.4). Radiotherapy combined with ipilimumab was associated with increased CD4+ and CD8+ICOS+ T cells. Increased CD8+ was significantly associated with PFS. When combined with ipilimumab at 10 mg/kg, the MTD of radiotherapy was 9 Gy. This combination of ipilimumab and radiotherapy appears to be associated with antitumor activity. Increased CD8+ was significantly associated with PFS. Thus, immune biomarkers may be useful for early response evaluation. NCT01557114.


How to measure the quality of the OSCE: A review of metrics - AMEE guide no. 49.
With an increasing use of criterion-based assessment techniques in both undergraduate and postgraduate healthcare programmes, there is a consequent need to ensure the quality and rigour of these assessments. The obvious question for those responsible for delivering assessment is how is this 'quality' measured, and what mechanisms might there be that allow improvements in assessment quality over time to be demonstrated? Whilst a small base of literature exists, few papers give more than one or two metrics as measures of quality in Objective Structured Clinical Examinations (OSCEs). In this guide, aimed at assessment practitioners, the authors aim to review the metrics that are available for measuring quality and indicate how a rounded picture of OSCE assessment quality may be constructed by using a variety of such measures, and also to consider which characteristics of the OSCE are appropriately judged by which measure(s). The authors will discuss the quality issues both at the individual station level and across the complete clinical assessment as a whole, using a series of 'worked examples' drawn from OSCE data sets from the authors' institution.


Triiodothyronine (T3) modulates hCG-regulated progesterone secretion, cAMP accumulation and DNA content in cultured human luteinized granulosa cells.
Ample clinical evidence indicates that women with thyroid disorders frequently exhibit menstrual disturbances and impaired fertility. In order to characterize the nature of thyroid hormone action in the ovary, the direct effects of triiodothyronine (T3) were investigated in vitro using a culture system of human luteinized granulosa cells. The presence of T3 receptors was also searched in such cells. The cell cultures were maintained in serum-free Ham's F-10 medium in the absence or presence of hCG, with or without graded doses of T3 (10(-11)-10(-7) M), and cell proliferation (assessed by DNA content) as well as cell function (cAMP accumulation and progesterone secretion) determined. T3 alone stimulated cell proliferation. hCG, on the other hand, was anti-mitogenic and T3 combined with hCG inhibited cell growth even further, reaching levels below those reached by either control or hCG alone. Exposure of cells to T3 markedly enhanced hCG-induced cAMP accumulation. Addition of 1-methyl-3-isobutylxanthine (MIX) abolished the cAMP-stimulatory effect elicited by T3, suggesting that the thyroid hormone may act, as MIX, by inhibiting phosphodiesterase. T3 was devoid of any influence on basal progesterone secretion, but inhibited hCG-induced secretion of the steroid. The effects of T3 are not accounted for by changes in cell number since the influence of thyroid hormone on cAMP and steroid secretion were expressed per microgram DNA.(ABSTRACT TRUNCATED AT 250 WORDS)


High level risky sexual behavior among persons living with HIV in the urban setting of the highest HIV prevalent areas in Ethiopia: Implications for interventions.
Data on the sexual behavior among people living with human immunodeficiency virus (PLHIV) dwelling at HIV prevalent setting located at the periphery of Ethiopia is lacking. Therefore, this study was designed to investigate sexual practice of patients following their antiretroviral therapy (ART) service and factors affecting their behavior. A facility based cross-sectional study design was employed to assess risky sexual practice and associated factors among HIV positive adults attending ART clinics in Gambella town, Southwest Ethiopia. Risky sexual practice is defined as a custom of getting in at least one of the following practices such as condom-unprotected sex with any partner, having two or more sexual partners and practicing casual sex in the last three months. A total of 352 randomly selected clients were interviewed by using a structured questionnaire. The multivariable logistic regression model was used to examine the association between covariates and the outcome variable. Majority of the study participants were engaged at least in one of the risky sexual practices (79.8% confidence interval (CI): 75.3% - 83.9%). The multivariable analysis found that the odds of risky sexual practice were higher among individuals who use substances ('khat' users (AOR: 3.82, 95%CI:1.30-11.22), smoke cigarette (AOR:4.90, 95%CI:1.19-12.60), consume alcohol (AOR: 2.59, 95%CI:1.28-5.21)); those who never discuss about safe sex with their partner/s (AOR: 2.21, 95%CI:1.16-4.21); those who have been in attachment for longer duration (more than four years) with their partner (AOR: 3.56, 95%CI: 1.32-9.62); and groups who desire to bear children in their future life (AOR: 3.15, 95%CI:1.40-7.04) as compared to their respective comparison groups. A significant number of participants were engaged at least in one of the risky sexual practices which potentially result in super infection by a new or/and drug resistant viral strain/s, and onward transmission of the virus. Thus, an HIV intervention program which focuses on the identified factors has to be implemented to mitigate risk of unsafe sexual behavior of this population group and move towards ending the HIV/Acquired Immunodeficiency Syndrome (AIDS) epidemic.


Integrated spatial health assessment of yellow perch (Perca flavescens) populations from the St. Lawrence River, Quebec, Canada) part A: physiological parameters and pathogen assessment.
A multi-disciplinary approach was used to evaluate the health of yellow perch (Perca flavescens) in the St. Lawrence River (Quebec, Canada), which is experiencing a severe population decline in the downstream portion of the river. Physiological parameters, liver alterations, trace metal concentrations, parasite prevalence and abundance, stable isotope composition, and the presence/absence of the viral hemorragic septicemia virus (VHSV) were evaluated in perch collected at six sites along the river: Lake St. François, Lake St. Louis (north and south), Beauregard Island, and Lake St. Pierre (north and south). Trace metal concentrations in surface water were higher in Lake St. Louis and downstream of a major urban wastewater treatment plant discharge, indicating that this effluent was a significant source of Cu, As, Ag, Zn, and Cd. Levels of Pb in surface water exceeded thresholds for the protection of aquatic life in Lake St. Louis and were negatively correlated with body condition index in this lake. In Lake St. Pierre, Cu, Ag, and Cd bioaccumulated significantly in perch liver and lower body condition index and greater liver damage were observed compared to upstream sites. Parasite analyses indicated a higher abundance of metacercariae of the trematodes Apophallus brevis and Diplostomum spp. in Lake St. Louis, and VHSV was not detected in the liver of yellow perch for all studied sites. Overall, results suggested that the global health of yellow perch from Lake St. Pierre is lower compared to upstream studied sites, which could contribute to the documented population collapse at this site.


Heart Rate Variability Is Associated with Motor Outcome 3-Months after Stroke.
The primary objective of this paper was to determine whether heart rate variability (HRV) acquired upon admission to inpatient rehabilitation is associated with motor outcome 3 months after stroke. The secondary objective of this paper was to determine whether HRV shows a strong association with the motor outcome 3 months after stroke in individuals with severe initial motor impairments. We recruited 13 patients with acute stroke from an acute inpatient rehabilitation hospital. A Holter monitor was placed upon admission and Fugl-Meyer Upper Extremity and Lower Extremity Subscales were used to assess the movement of the affected upper and lower extremities 3 months after admission. The standard deviation of R-R intervals was used to quantify HRV. A Spearman rank correlation revealed a strong positive and significant correlation between HRV upon admission and movement of the affected upper extremity (r = .70, P = .01) and affected lower extremity (r = .60, P = .03) at 3 months. For patients with severe initial motor impairments, HRV showed a strong positive association with the movement of the affected upper (r = .61, P = .04) and lower (r = .70, P = .04) extremities at 3 months. HRV is strongly associated with motor outcome after stroke and provides a promising marker to explore the mechanisms associated with motor recovery after stroke.


Enhanced removal of naproxen and carbamazepine from wastewater using a novel countercurrent seepage bioreactor immobilized with Phanerochaete chrysosporium under non-sterile conditions.
A countercurrent seepage bioreactor immobilized with Phanerochaete chrysosporium was continuously operated under non-sterile conditions to treat a synthetic wastewater spiked with naproxen and carbamazepine (1000μg/L each) for 165days. There were no serious bacterial contaminations occurred during the operational period. Naproxen was always removed to the undetectable level regardless of the experimental conditions, while the average removal efficiency for carbamazepine, a well-known recalcitrant pharmaceutically active compound, reached around 80%. The excellent removal performance was mainly attributed to the application of countercurrent seepage mode and the cardhouse fabric of the carriers, which provided the high efficiency in the transfer of oxygen and nutrients inside the bioreactor. From the fungal immobilization combined with the temperature adjustment, the fungal activity including the enzyme production was protected as well as the bacterial contamination inside the reactor was suppressed effectively.


The role of autophagy dysregulation in manganese-induced dopaminergic neurodegeneration.
The etiological role of dysregulated autophagy in neurodegenerative diseases has been a subject of intense investigation. While manganese (Mn) is known to cause dopaminergic (DAergic) neurodegeneration, it has yet to be determined whether the dysregulation of autophagy plays a role in Mn-induced neuronal injury. In this study, we investigated the effect of Mn on autophagy in a rat model of manganism, a neurodegenerative disease associated with excessive exposure to Mn. After a single intrastriatal injection of Mn, the short- (4-12 h) and long-term (1-28 days) effect of Mn on DAergic neurons and autophagy were examined. Marked reduction in the number of TH-immunoreactive neurons in the substantia nigra pars compacta (SNpc) as well as TH protein expression, and a significant increase of apomorphine-induced rotations were observed in rats after Mn injection. Manganese also induced the down-regulation of dopamine levels and D1 dopamine receptor expression. In addition, autophagy was dysregulated and inhibited, as evidenced by increased number of abnormal lysosomes, decreased protein expression of Beclin1, and decreased ratio of microtubule-associated protein 1 light chain 3 (LC3) II over LC3 I, concomitant with activated mammalian target of rapamycin (mTOR)/p70 ribosomal protein S6 kinase (p70s6k) signaling. In contrast, in the early phase of Mn exposure, the level of autophagy was not be suppressed but compensatorily activated. Although the morphology of the DAergic neuron was intact in the early phase, Mn caused a significant decrease in TH-immunoreactivity and a significant increase in apomorphine-induced rotations in the presence of wortmannin, an inhibitor of autophagy. Taken together, these results demonstrate, for the first time, that autophagy may play a protective role against Mn-induced neuronal damage, whilst dysregulation of autophagy at later phases may mediate DAergic neurodegeneration.


Declining tree growth resilience mediates subsequent forest mortality in the US Mountain West.
Climate change-triggered forest die-off is an increasing threat to global forests and carbon sequestration but remains extremely challenging to predict. Tree growth resilience metrics have been proposed as measurable proxies of tree susceptibility to mortality. However, it remains unclear whether tree growth resilience can improve predictions of stand-level mortality. Here, we use an extensive tree-ring dataset collected at ~3000 permanent forest inventory plots, spanning 13 dominant species across the US Mountain West, where forests have experienced strong drought and extensive die-off has been observed in the past two decades, to test the hypothesis that tree growth resilience to drought can explain and improve predictions of observed stand-level mortality. We found substantial increases in growth variability and temporal autocorrelation as well declining drought resistance and resilience for a number of species over the second half of the 20th century. Declining resilience and low tree growth were strongly associated with cross- and within-species patterns of mortality. Resilience metrics had similar explicative power compared to climate and stand structure, but the covariance structure among predictors implied that the effect of tree resilience on mortality could partially be explained by stand and climate variables. We conclude that tree growth resilience offers highly valuable insights on tree physiology by integrating the effect of stressors on forest mortality but may have only moderate potential to improve large-scale projections of forest die-off under climate change.


Label-free quantitative proteomic analysis of insect larval and metamorphic molts.
Molting is an essential biological process occurring characteristic times throughout the life cycle of holometabolous insects. However, it is not clear how insects determine the direction of molting to remain status quo or to initiate metamorphosis. To explore the functional factors that determine the direction of molts, liquid chromatography-mass spectrometry was used to identify the molecules involved in larval and metamorphic molting, and the differentially expressed proteins (DEPs) were compared in the two processes. There were 321 and 1140 DEPs identified in larval and metamorphic molting process, respectively. Bioinformatics analyses show that the amino sugar pathway was up-regulated in both processes. The up-regulated protease contributed to the metamorphosis. In addition, several proteins with different expression patterns in larval-larval and larval-pupal transitions, including Endochitinase, GRIM-19 (Genes associated with retinoid-IFN-induced mortality-19), IDE (Insulin-degrading enzyme), Sorcin (Soluble resistance related calcium binding protein), OBP (Odorant-binding protein-2 precursor), TRAP1(Tumor necrosis factor receptor associated protein-1), etc., were further identified by parallel reaction monitoring, which may play diverse functions in larval-larval and larval-pupal transitions. These results provide a proteomic insight into molecules involved in larval and metamorphic molts, and will likely improve the current understanding of determination of direction of molts.


Heme-solvent coupling: a Mössbauer study of myoglobin in sucrose.
The Mössbauer effect of 57Fe-enriched samples was used to investigate the coupling of 80% sucrose/water, a protein-stabilizing solvent, to vibrational and diffusive modes of the heme iron of CO-myoglobin. For comparison we also determined the Mössbauer spectra of K4 57Fe (CN)6 (potassium ferrocyanide, PFC), where the iron is fully exposed in the same solvent. The temperature dependence of the Mössbauer parameters derived for the two samples proved to be remarkably similar, indicative of a strong coupling of the main heme displacements to the viscoelastic relaxation of the solvent. We show that CO escape out of the heme pocket couples to the same type of fluctuations, whereas intramolecular bond formation involves solvent-decoupled heme deformation modes that are less prominent in the Mössbauer spectrum. With respect to other solvents, however, sucrose shows a reduced viscosity effect on heme displacements and the kinetics of ligand binding due to preferential hydration of the protein. This result confirms thermodynamic predictions of the stabilizing action of sucrose by a dynamic method.


A method to identify acoustic reverberation in multilayered homogeneous media.
The presence of reverberation is a source of artifacts that can hinder the analysis of ultrasound signals and images. Besides compromising image generation, these artifacts can introduce errors in the quantitative parameter estimation in fields such as material and biological tissue characterization. A method that allows the separation between the first reflection on an interface and all the other reflections from the same interface (reverberation) could improve the quality of these images as well as the precision and accuracy in quantitative results. This work presents an algorithm for the identification of reverberating echoes in multilayered media, based on the comparison of their power spectra (estimated via FFT), through a least mean square approach, and on the temporal relationship among them. It considers that the global effect from attenuation, reflection and transmission coefficients for each layer causes spectral differences that could differentiate echoes that pass through one layer or another. The results of 10 simulations and of 60 experiments, carried out with 6 different phantoms (10 experiments with each one) are presented and discussed. It was found that the algorithm provided a correct identification for 85% of the simulated and 86.6% of the experimental echoes collected from the 60 experiments.


ST waveform analysis versus cardiotocography alone for intrapartum fetal monitoring: a systematic review and meta-analysis of randomized trials.
ST waveform analysis was introduced to reduce metabolic acidosis at birth and avoid unnecessary operative deliveries relative to conventional cardiotocography. Our objective was to quantify the efficacy of ST waveform analysis vs. cardiotocography and assess the quality of the evidence using the Grading of Recommendations Assessment, Development and Evaluation tool. We identified randomized controlled trials through systematic literature searches and assessed included studies for risk of bias. Meta-analyses were performed, calculating pooled risk ratio or peto odds ratio. We performed post hoc trial sequential analyses for selected outcomes to assess the risk of false-positive results and the need for additional studies. Six randomized controlled trials were included in the meta-analysis. ST waveform analysis was not associated with a reduction in operative deliveries due to fetal distress, but we observed a significantly lower rate of metabolic acidosis (peto odds ratio 0.64; 95% confidence interval 0.46-0.88). Accordingly, 401 women need to be monitored with ST waveform analysis to prevent one case of metabolic acidosis. No statistically significant effects were observed in other fetal or neonatal outcomes, except from fetal blood sampling (risk ratio 0.59; 95% confidence interval 0.45-0.79) and a minor reduction in the number of operative vaginal deliveries for all indications (risk ratio 0.92; 95% confidence interval 0.86-0.99). The quality of the evidence was high to moderate. Absolute effects of ST waveform analysis were minor, and the clinical significance of the observed reduction in metabolic acidosis is questioned. There is not enough evidence to justify the use of ST waveform analysis in contemporary obstetrics.


Population cytogenetics of Chironomus circumdatus Kieffer, 1921 (Diptera, Chironomidae) from Thailand.
The objectives of this study were to explore cytogenetic variation and the role of chromosomal change on local adaptation and genetic differentiation of Chironomus circumdatus Kieffer from Thailand. A total of 1,505 larvae from 24 populations were examined cytologically. Twelve chromosomal inversions were found and most of these (9 of 12) were rare inversions. All populations were in Hardy-Weinberg equilibrium. Significant association (P < 0.001) between the A2 and B5 inversions was detected in one population. Population genetic structure analysis indicated significant genetic differentiation between populations (F (ST) = 0.037, P < 0.001). Geographic distance was the principal factor limiting gene flow between populations. Nei's modified genetic distance (D (A)) between populations ranged from 0.001 to 0.011 with an average of 0.003. An UPGMA population phenogram depicting relationship between populations based on D (A) values revealed three groups of populations, group I, II and III each characterized by different inversions/inversion frequencies. Significant correlation of inversion C3 and water temperature suggested that this inversion might have a role to play on adaptation to high temperature habitat. However, if detection of significant population subdivision and relationship between genetic and geographic distance are taken into account, relationship between C3 and water temperature will also be due to the effect of migration/drift alone without the effect of selection.


Mixture-based heterocyclic combinatorial positional scanning libraries: discovery of bicyclic guanidines having potent antifungal activities against Candida albicans and Cryptococcus neoformans.
A mixture-based synthetic combinatorial library of more than 100,000 bicyclic guanidines was generated in a positional scanning format and assayed for activity against Candida albicans. Potent individual bicyclic guanidines were directly identified following the screening of the library. Time-kill curve studies indicated bactericidal activities for the individual bicyclic guanidines. These compounds also showed potent activity against Cryptococcus neoformans. These studies demonstrate the value of using mixture-based combinatorial positional scanning libraries made up of heterocyclic compounds for the rapid identification of novel classes of antifungal compounds.


Microbial Phototrophic, Heterotrophic, and Diazotrophic Activities Associated with Aggregates in the Permanent Ice Cover of Lake Bonney, Antarctica.
Abstract The McMurdo Dry Valley lakes, Antarctica, one of the Earth's southernmost ecosystems containing liquid water, harbor some of the most environmentally extreme (cold, nutrient-deprived) conditions on the planet. Lake Bonney has a permanent ice cover that supports a unique microbial habitat, provided by soil particles blown onto the lake surface from the surrounding, ice-free valley floor. During continuous sunlight summers (Nov.-Feb.), the dark soil particles are heated by solar radiation and melt their way into the ice matrix. Layers and patches of aggregates and liquid water are formed. Aggregates contain a complex cyanobacterial-bacterial community, concurrently conducting photosynthesis (CO2 fixation), nitrogen (N2) fixation, decomposition, and biogeochemical zonation needed to complete essential nutrient cycles. Aggregate-associated CO2- and N2-fixation rates were low and confined to liquid water (i.e., no detectable activities in the ice phase). CO2 fixation was mediated by cyanobacteria; both cyanobacteria and eubacteria appeared responsible for N2 fixation. CO2 fixation was stimulated primarily by nitrogen (NO3-), but also by phosphorus (PO43-). PO43- and iron (FeCl3 + EDTA) enrichment stimulated of N2 fixation. Microautoradiographic and physiological studies indicate a morphologically and metabolically diverse microbial community, exhibiting different cell-specific photosynthetic and heterotrophic activities. The microbial community is involved in physical (particle aggregation) and chemical (establishing redox gradients) modification of a nutrient- and organic matter-enriched microbial "oasis," embedded in the desertlike (i.e., nutrient depleted) lake ice cover. Aggregate-associated production and nutrient cycling represent microbial self-sustenance in a microenvironment supporting "life at the edge," as it is known on Earth.


Serrated Colorectal Cancer: The Road Less Travelled?
Studies of colorectal cancer (CRC) originating through the conventional adenoma-carcinoma sequence have provided insight into the molecular mechanisms controlling its initiation and progression. Less is known about the alternative 'serrated' pathway, which has been associated with BRAF mutation and microsatellite instability. Recent transcriptomics approaches to classify human CRC revealed that mesenchymal and/or desmoplastic features combined with an immunosuppressive microenvironment are key determinants of CRC with the poorest prognosis. Importantly, these aggressive CRCs harbor the characteristics of serrated tumors, suggesting that initiation through this alternative pathway determines how aggressive the CRC becomes. Here, we review recent evidence on how serrated carcinogenesis contributes to the subtype of CRC with the poorest prognosis.


Embryonic toxicity of 3,4-dichloroaniline (3,4-DCA) on Javanese medaka (Oryzias javanicus Bleeker, 1854).
Early-life exposure to toxic chemicals causes irreversible morphological and physiological abnormalities that may last for a lifetime. The present study aimed to determine the toxicity effect of 3,4-Dichloroaniline (3,4-DCA) on Javanese medaka (Oryzias javanicus) embryos. Healthy embryos were exposed to various 3,4-DCA concentrations for acute toxicity (5, 10, 25, 50, and 100 mg.L-1) and sublethal toxicity (0.10, 0.50, 1.25, 2.50, and 5.00 mg.L-1) for 96 h and 20 days respectively. Acute toxicity test revealed that the median lethal concentration (96h-LC50) was 32.87 mg.L-1 (95 % CI = 27.90-38.74, R2 = 0.95). Sublethal exposure revealed that 1.25 mg.L-1 at 3 days post-exposure (3 dpe) has a significant lower heartrate (120 ± 12.3 beats/min., p < 0.01), while at 7 dpe those exposed to 5 mg.L-1 (141.8 ± 8.3 beats/min) had significantly (p < 0.01) lower heart rate compared to other treatments. Likewise, at 13 dpe, 5.00 mg.L-1 (110.4 ± 17.3 beats/min) and 2.5 mg.L-1 (130.4 ± 8.3 beats/min) were significantly lower (p < 0.001) compared to control. None of the embryos in 5.00 mg.L-1 and 2.50 mg.L-1 treatment groups survived at the end of the experiment. The results indicated a concentration-dependent response. The lowest observed effect concentration (LOEC) that exerted developmental deformities was 0.5 mg.L-1. Javanese medaka embryo have low sensitivity to acute toxicity of 3,4-DCA, but developmental abnormalities at sublethal concentrations were observed.


Barriers to knowledge production, knowledge translation, and urban health policy change: ideological, economic, and political considerations.
In this paper, we consider social forces that affect the processes of both knowledge production and knowledge translation in relation to urban health research. First, we briefly review our conceptual model, derived from a social-conflict framework, to outline how unequal power relations and health inequalities are causally linked. Second, we critically discuss ideological, political, and economic barriers that exist within academia that affect knowledge production related to urban health and health inequalities. Third, we broaden the scope of our analysis to examine how the ideological, political, and economic environment beyond the academy creates barriers to health equity policy making. We conclude with some key questions about the role that knowledge translation can possibly play in light of these constraints on research and policy for urban health.


POST-TREATMENT PREDICTION OF OPTICAL COHERENCE TOMOGRAPHY USING A CONDITIONAL GENERATIVE ADVERSARIAL NETWORK IN AGE-RELATED MACULAR DEGENERATION.
To develop a deep learning model to generate posttreatment optical coherence tomography (OCT) images of neovascular age-related macular degeneration. Two hundred ninety-eight patients with neovascular age-related macular degeneration were included. The conditional generative adversarial network was trained using 15,183 augmented paired OCT B-scan images obtained from 723 scans of 241 patients at baseline and 1 month after 3 loading doses of an anti-vascular endothelial growth factor treatment. The network was also trained using baseline fluorescein angiography (FA) or indocyanine green angiography (ICGA) images together with baseline OCT images. A test set of 150 images of 50 eyes was used to evaluate its ability to predict the presence of intraretinal fluid, subretinal fluid, PED, and subretinal hyperreflective material. Posttreatment OCT images were compared with images generated from baseline OCT with or without FA and indocyanine green angiography images. The predicted images inferred from baseline OCT images achieved an acceptable accuracy, specificity, and negative predictive value for four lesions (range: 77.0-91.9, 94.1-95.1, and 54.7-96.5%, respectively). The addition of both FA and indocyanine green angiography images improved the accuracy, specificity, and negative predictive value (range: 80.7-96.3, 97.3-99.0, and 59.0-98.3%, respectively). A conditional generative adversarial network is able to generate posttreatment OCT images from baseline OCT, FA, and indocyanine green angiography images.


The effects of feeding immunostimulant β-glucan on the immune response of Pangasianodon hypophthalmus.
Immunostimulants are food additives used by the aquaculture industry to enhance the immune response of fish, and although β-glucans are now commonly used for this purpose in aquaculture, little is known about their effects on the immune response of Pangasianodon hypophthalmus. Thus, a variety of immune parameters (e.g. phagocytosis, respiratory burst, lysozyme, complement, peroxidase, total protein, total anti-protease, total IgM, natural antibody titres, and specific IgM titres) was examined in this species after feeding fish with a basal control diet or diets supplemented with 0.05, 0.1, or 0.2 g/kg fungal-derived β-glucan or 0.1% commercial yeast-derived β-glucan, as a positive control diet, for a period of four weeks. The effect of the glucans on disease resistance was then evaluated by experimentally infecting the fish with Edwardsiella ictaluri by immersion and mortalities monitored for 14 days. Samples were collected from fish for analysis at 0, 1, 3, 7, 14, 21 and 28 days post-feeding (dpf), and also at 14 days post infection (dpi). The lowest dose of fungal-derived β-glucan (0.05%) appeared insufficient to effectively stimulate the immune response of the fish, while those fed with the two highest levels of fungal-derived β-glucan had enhanced immune responses compared to the control group. Significantly elevated levels of respiratory burst activity on all days examined (P < 0.05) and lysozyme activity on 7 dpf were found in the group fed 0.2% fungal-derived β-glucan, while plasma anti-protease activity was significantly enhanced (P < 0.05) by 21 dpf, natural antibody titres by 3 dpf and complement activity by 7 dpf and also at 14 dpi in the group fed 0.1% fungal-derived β-glucan. No statistical differences was seen in the level of mortalities between the dietary groups, although the group fed with the control diet had the highest level of mortalities and the groups fed with commercial yeast-derived β-glucan and 0.2% fungal-derived β-glucan the lowest.


Adsorption of Bismark Brown dye on activated carbons prepared from rubberwood sawdust (Hevea brasiliensis) using different activation methods.
Hevea brasiliensis or rubberwood tree, as it is commonly known finds limited use once the latex has been tapped. The sawdust of this tree is chosen to ascertain it viability as a precursor for activation. The carbons thus obtained were characterized in terms of iodine, methylene blue number and surface area. The best carbon in each method was utilized to study the adsorption of Bismark Brown, a dye used in the leather industry. Adsorption equilibrium studies were carried out with the synthetic solutions of the dye, at room temperature (298 K). Equilibrium data are fitted with the Langmuir and the Freundlich isotherms models for the system. The effects of contact time, adsorbent dosage and initial dye concentrations on sorption capacity were carried out. Excellent adsorption capacities of 2000 and 1111 mg g(-1) were obtained for steam and chemical followed by steam-activated carbons, respectively. Pilot-plant experimental studies have been performed using packed-bed column with different feed concentrations, flow rates and bed heights, to evaluate sorption of Bismark Brown on steam-activated carbon. Bed depth service time (BDST) design model have been used to analyze the data.


Effectiveness of short vs long-course perioperative antibiotics in lung transplant recipients with donor positive respiratory cultures.
Lung transplant recipients are at increased risk for infection in the early postoperative phase, thus perioperative antibiotics are employed. This retrospective study evaluated the efficacy of short- vs long-course perioperative antibiotics in lung transplant patients. Lung transplant patients with donor positive cultures between August 2013 and September 2019 were evaluated, excluding those with cystic fibrosis, death within 14 days and re-transplants. The primary outcome was 30-day freedom from donor-derived respiratory infection. A total of 147 patients were included (57 short vs 90 long-course). Median perioperative antibiotic duration was 6 days in the short-course vs 14 days in the long-course group (P < .0001). Thirty-day freedom from donor-derived respiratory infection was present in 56 (98%) patients in the short-course vs 85 (94%) patients in the long-course group (P = .41). There was no difference in development of Clostridioides difficile infections (P = .41), while cumulative ventilator time and time to post-op extubation were longer in the long-course group (P = .001 and .004, respectively). Among lung transplant recipients with positive donor respiratory cultures, short-course perioperative antibiotics were as effective as long-course antibiotics in preventing donor-derived bacterial respiratory infections.


Central place foraging by beavers (Castor canadensis): a test of foraging predictions and the impact of selective feeding on the growth form of cottonwoods (Populus fremontii).
Several predictions of central place foraging theory were tested. As predicted, beavers foraged more selectively at increasing distance from the central place. With increasing distance from the river's edge, beavers cut fewer branches and deleted small branches from their diet. Large branches were favored at all distances, which differs from patterns observed in previous studies of beaver foraging. This difference, however, is expected and supports Schoener's (1979) predictions which are based on differences between provisioning costs and item size.The selective harvesting of branches predicted by foraging theory affects plant growth form and may influence plant reproductive patterns. High rates of branch removal caused cottonwoods to develop a shrubby architecture. The importance of selective branch choice by beavers on patterns of cottonwood reproduction (i.e., delayed sexual maturity and induced cloning) is discussed.


Gd(minus)Matam, an African glucose-6-phosphate dehydrogenase variant with enzyme deficiency. Biochemical and immunological properties in various hemopoietic tissues.
Two unrelated Senegalese patients, both native of the Matam province, were found to have the same deficient G6PD variant. One has no hematological history, the other had several induced acute hemolytic episodes. The deficiency was almost complete in red blood cells and 20-30 percent of the normal level in leukocytes and platelets; in leukocytes the deficiency was due to a decrease in the molecular specific activity of the enzyme to which a molecular instability was added, explaining the greater deficiency in red blood cells. The electrophoretic mobility was slightly fast in leukocytes and platelets but normal in red blood cells. This pattern was confirmed by electrofocusing in ampholine-acrylamide gel. From a kinetic point of view, these enzymes were characterized by a lowered Km (G6P) (13 to 20 muM) a normal Km (NADP+), a Ki (NADPH) increased about twice, a thermal instability, a biphasic pH curve and an increased activation energy (15 kcal/mole). The polymorphonuclear cells were functionally strictly normal: engulfment, nitroblue tetrazolium (NBT) reduction test, induced iodination, and oxygen consumption were normal. The authors discuss the importance of post-synthetic modifications of the muted enzymes and their repercussions on the enzyme characteristics.


Simultaneous Torsion of a Uterine Horn and Spleen in a Pregnant Two-Year-Old Crossbreed Bitch: A Case Report.
A pregnant 2-year-old mixed-breed dog was admitted with a 2-day history of lethargy, anorexia and painful abdominal distension. Clinical manifestations were unspecific and mainly suggested hypovolemic shock. Physical examination, ultrasonographic evaluation and radiographs confirmed mid-late pregnancy and evident signs of fetal death and peritoneal effusion. An exploratory laparotomy was immediately initiated which revealed torsion of the right gravid uterine horn over the left one and, simultaneously, the unexpected presence of splenic torsion and rupture, with a stretched omentum covering and exerting tension on the spleen's cranial pole. Histopathology revealed hemorrhagic infarction of the uterus and spleen. The animal recovered uneventfully and was later submitted to a preventive gastropexy.


Anesthetic management of a parturient with hypermobility phenotype but possible vascular genotype Ehlers-Danlos syndrome.
We describe a case of a term parturient previously clinically diagnosed with hypermobility type Ehlers-Danlos syndrome (EDS) but later diagnosed with a genotype that may be associated with vascular type EDS. This 26-yr-old primigravida had been clinically diagnosed with hypermobility type EDS prior to her pregnancy. Nevertheless, subsequent genetic testing during pregnancy revealed a heterozygous variant of unknown significance in the COL3A1 gene causative for vascular type EDS. A multidisciplinary decision was made to prioritize the phenotype rather than the genotype in regard to clinical management of labour. An induced labour was planned with instrument-assisted vaginal delivery. We elected to proceed with placement of an epidural catheter for provision of labour analgesia and anesthesia during delivery. Both maternal and neonatal outcomes were excellent. The risk of severe morbidity and mortality in parturients with vascular EDS has warranted recommendations for modified management of labour, particularly regarding mode and timing of delivery. Nevertheless, a multidisciplinary approach and consideration of phenotype rather than genotype alone were instrumental in the successful management of this patient. Genetic testing of patients who display features of EDS and/or who have a positive family history of the disease is important in the preparation for labour and delivery. In the absence of convincing signs of vascular EDS and a negative family history, it may be rational to offer certain parturients neuraxial anesthesia and a trial of vaginal labour.


Inositol tetrakisphosphate (IP4)- and inositol triphosphate (IP3)-dependent Ca2+ influx in cortical neuronal nuclei of newborn piglets following graded hypoxia.
Previous studies have shown that hypoxia results in a modification of the binding characteristics of the neuronal nuclear membrane inositol tetrakisphosphate (IP4) and inositol triphosphate (IP3) receptors. The present study tests the hypothesis that hypoxia-induced modification of the IP4 and IP3 receptors results in increased IP4 and IP3 dependent Ca2+ influx in neuronal nuclei as a function of the degree of cerebral tissue hypoxia in newborn piglets. Studies were performed in piglets, 3-5 days old, divided into normoxic (N = 5) and hypoxic (N = 6) groups. The hypoxic group was exposed to decreased FiO2 ranging from 0.15 to 0.05 for 1 h. Brain tissue hypoxia was documented biochemically by determining ATP and phosphocreatine (PCr) levels. Neuronal nuclei were isolated and 45Ca2+ influx was determined in a medium containing 50 mM Tris buffer (pH 7.4), neuronal nuclei (150 microg protein), 1 microM 45Ca2+, with or without 10 microM IP4 or IP3. In normoxic and hypoxic groups, ATP levels were 4.27 +/- 0.80 and 1.40 +/- 0.69 micromoles/g brain, respectively, P < .001 (ranging from 4.78 to 0.82). PCr levels were 3.40 +/- 0.99 and 0.91 +/- 0.57 micromoles/g brain, respectively, P < .001 (raning from 4.07 to 0.60). During hypoxia, IP4-dependent intranuclear 45Ca2+ influx increased from 3.39 +/- 0.64 in normoxic nuclei to 13.30 +/- 2.18 pM/mg protein in hypoxic nuclei (P < .01). There was an inverse correlation between the 45Ca2+ influx in neuronal nuclei and the levels of cerebral tissue ATP (r = 0.83) and PCr (r = 0.85). Similarly, IP3-dependent intranuclear 45Ca2+ influx increased from 2.26 +/- 0.38 pmoles/mg protein in normoxic nuclei to 11.12 +/- 1.65 pmoles/mg protein in hypoxic nuclei and showed an inverse correlation between 45Ca2+ influx in neuronal nuclei and the levels of cerebral tissue ATP (r = 0.86) and PCr (r = 0.71). The data demonstrate that there is an IP4- as well as IP3-dependent increase in nuclear Ca2+ influx with increasing cerebral tissue hypoxia, suggesting a hypoxia-induced modification of the nuclear membrane IP4 and IP3 receptors. We propose that there is a specific level of tissue hypoxia that results in a critical increase of intranuclear Ca2+ that leads to altered transcription of apoptotic genes and activation of nuclear endonucleases resulting in hypoxia-induced programmed neuronal death.


Descriptive epidemiology of cancer of unknown primary site in Scotland, 1961-2010.
Cancers of unknown primary site (CUP) pose problems for diagnosis, treatment, and accurate prediction of prognosis. However, there are limited published data describing the epidemiology of this disease entity. Our aim was to describe the epidemiology of CUP in Scotland. Anonymised data, covering the period 1961-2010, were extracted from the Scottish Cancer Registry database, based on the following ICD-10 diagnostic codes: C26.0, C26.8, C26.9, C39, and C76-C80. Age-standardised incidence rates were calculated by direct standardisation to the World Standard Population. Estimates of observed survival were calculated by the Kaplan-Meier method. Between 1961 and 2010, there were 50,941 registrations of CUP, representing 3.9% of all registrations of invasive cancers. Age-standardised rates increased to a peak in the early to mid-1990s, followed by a steeper decrease in rates. During 2001-2010, age-standardised rates of CUP were higher in the most compared with the least deprived fifth of the population. Observed survival was marginally higher in patients diagnosed during 2001-2010 (median 5.6 weeks) compared with those diagnosed in the previous two decades. During the most recent decade, survival decreased with age at diagnosis, and was higher in patients with squamous cell carcinoma and with lymph node metastases. Patterns of CUP in Scotland are largely consistent with those reported from the few other countries that have published data. However, in comparing studies, it is important to note that there is heterogeneity in terms of definition of CUP, as well as calendar period of diagnosis or death. Variation in the definition of CUP between different epidemiological studies suggests that there would be merit in seeking international agreement on guidelines for the registration of CUP as well as a standard grouping of diagnostic codes for analysis.


Differentiation of human embryonic stem cells to regional specific neural precursors in chemically defined medium conditions.
Human embryonic stem cells (hESC) provide a unique model to study early events in human development. The hESC-derived cells can potentially be used to replace or restore different tissues including neuronal that have been damaged by disease or injury. The cells of two different hESC lines were converted to neural rosettes using adherent and chemically defined conditions. The progenitor cells were exposed to retinoic acid (RA) or to human recombinant basic fibroblast growth factor (bFGF) in the late phase of the rosette formation. Exposing the progenitor cells to RA suppressed differentiation to rostral forebrain dopamine neural lineage and promoted that of spinal neural tissue including motor neurons. The functional characteristics of these differentiated neuronal precursors under both, rostral (bFGF) and caudalizing (RA) signals were confirmed by patch clamp analysis. These findings suggest that our differentiation protocol has the capacity to generate region-specific and electrophysiologically active neurons under in vitro conditions without embryoid body formation, co-culture with stromal cells and without presence of cells of mesodermal or endodermal lineages.


Integrating Pediatric Care And Taking On Risk To Improve Health.
In Delaware, Nemours Children's Health System wants to get paid for keeping children healthy.


Comparative Analysis of Clinical Outcomes for COVID-19 and Influenza among Cardiac Transplant Recipients in the United States.
COVID-19 infections can lead to worse outcomes in an immunocompromised population with multiple comorbidities, e.g., heart transplant patients. We used the National Inpatient Sample database to compare heart transplant outcomes in patients with COVID-19 vs. influenza. A total of 2460 patients were included in this study: heart transplant with COVID-19 (n = 1155, 47.0%) and heart transplant with influenza (n = 1305, 53.0%) with the primary outcome of in-hospital mortality. In-hospital mortality (n = 120) was significantly higher for heart transplant patients infected with COVID-19 compared to those infected with influenza (9.5% vs. 0.8%, adjusted OR: 51.6 [95% CI 4.3-615.9], p = 0.002) along with significantly higher rates of mechanical ventilation, acute heart failure, ventricular arrhythmias, and higher mean total hospitalization cost compared to the influenza group. More studies are needed on the role of vaccination and treatment to improve outcomes in this vulnerable population.


Effects of remifentanil on esophageal motility: a double-blind, randomized, cross-over study in healthy volunteers.
Recent studies have shown that remifentanil increases the risk of aspiration and induces subjective swallowing difficulties. The mechanisms are not completely understood. Here, we investigated whether remifentanil impairs esophageal motility and hypothesized that this is one possible underlying mechanism. Naloxone was used to evaluate whether the effects of remifentanil are mediated through opioid receptors. We also examined subjective swallowing difficulties and the influence of metoclopramide on remifentanil-induced effects. Fourteen healthy volunteers participated in a double-blind, randomized, cross-over trial at the University Hospital in Örebro, Sweden. They were studied on two different occasions, during which they were randomly assigned to receive either naloxone given as a bolus of 6 μg/kg followed by an infusion of 0.1 μg/kg/min, or saline 5 min before target-controlled infusions of remifentanil at three target-site concentrations: 1, 2, and 3 ng/ml. On both occasions, 0.2 mg/kg metoclopramide was given before the final measurement. Five swallows were performed during each measuring condition, and the metrics defining esophageal motility were measured by high-resolution manometry. Outcomes were differences in the metrics at baseline vs. during remifentanil infusion, with naloxone vs. placebo, and with remifentanil before and after metoclopramide administration. Differences in swallowing difficulties were also recorded. Remifentanil decreased swallow-evoked esophagogastric junction relaxation and the latency time of esophageal peristalsis. There were no significant effects of naloxone or metoclopramide on remifentanil-induced effects, and we detected no differences in swallowing difficulties. Remifentanil induces dysfunction of esophageal motility; this may contribute to the elevated risk of regurgitation and aspiration.


Family members of people with disabilities' explicit and implicit disability attitudes.
Family members of people with disabilities may hold a unique position in that they may both internalize and reinforce ableism, and work to fight it through empowerment and resistance, making them a useful group for study to elicit understandings of disability and disability attitudes. The aim of this study was to explore the explicit (conscious) and implicit (unconscious) attitudes toward people with disabilities by family members of persons with disabilities. To do so, we analyzed secondary data from 180,701 family members, comparing their explicit and implicit disability attitudes and examining correlates with attitudes. Findings from our study suggest that although family members do not consciously believe they have negative attitudes, they unconsciously prefer nondisabled people. More work is necessary to reduce prominent and systemic negative attitudes about disability as most family members still had negative attitudes about people with disabilities. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


GD2 CAR T cells against human glioblastoma.
Glioblastoma is the most malignant primary brain tumor and is still in need of effective medical treatment. We isolated patient-derived glioblastoma cells showing high GD2 antigen expression representing a potential target for CAR T strategy. Data highlighted a robust GD2 CAR antitumor potential in 2D and 3D glioblastoma models associated with a significant and CAR T-restricted increase of selected cytokines. Interestingly, immunosuppressant TGF β1, expressed in all co-cultures, did not influence antitumor activity. The orthotopic NOD/SCID models using primary glioblastoma cells reproduced human histopathological features. Considering still-conflicting data on the delivery route for targeting brain tumors, we compared intracerebral versus intravenous CAR T injections. We report that the intracerebral route significantly increased the length of survival time in a dose-dependent manner, without any side effects. Collectively, the proposed anti-GD2 CAR can counteract human glioblastoma potentially opening a new therapeutic option for a still incurable cancer.


Stage-specific detection of a DNA-binding protein for the storage protein gene of Sarcophaga peregrina.
A nuclear extract of fat body prepared from third instar larvae of Sarcophaga peregrina (flesh fly) was found to contain a DNA-binding protein that specifically bound to 5'-upstream region of the storage protein (arylphorin) gene. This protein was found only in larvae harvested 46 h after larval emergence, indicating that its appearance was strictly regulated by the developmental stage of the insect. Since the storage protein gene is actively transcribed in the fat body at this stage, this protein is probably a specific transcription factor for the storage protein gene. DNase I footprinting analysis showed that the nucleotide sequence of the binding site of this protein is ACCACAACA, which is located at residues -247 to -255 upstream of the +1 site. Results indicated that formation of the DNA-protein complex required Zn2+.


Causal roles of prefrontal cortex during spontaneous perceptual switching are determined by brain state dynamics.
The prefrontal cortex (PFC) is thought to orchestrate cognitive dynamics. However, in tests of bistable visual perception, no direct evidence supporting such presumable causal roles of the PFC has been reported except for a recent work. Here, using a novel brain-state-dependent neural stimulation system, we identified causal effects on percept dynamics in three PFC activities-right frontal eye fields, dorsolateral PFC (DLPFC), and inferior frontal cortex (IFC). The causality is behaviourally detectable only when we track brain state dynamics and modulate the PFC activity in brain-state-/state-history-dependent manners. The behavioural effects are underpinned by transient neural changes in the brain state dynamics, and such neural effects are quantitatively explainable by structural transformations of the hypothetical energy landscapes. Moreover, these findings indicate distinct functions of the three PFC areas: in particular, the DLPFC enhances the integration of two PFC-active brain states, whereas IFC promotes the functional segregation between them. This work resolves the controversy over the PFC roles in spontaneous perceptual switching and underlines brain state dynamics in fine investigations of brain-behaviour causality.


Complications of therapeutic plasma exchange: a prospective study of 1,727 procedures.
The type and number of complications was prospectively examined in 1,727 successive TPE treatments in 174 patients over 66 months at a single center. Most treatments were prescribed for thrombotic thrombocytopenic purpura (TTP; 42%), recurrent focal segmental glomerulosclerosis (FSGS; 22%), or myasthenia gravis (MG; 13%). About 57% of treatments used albumin-saline as the replacement solution and 43% used fresh-frozen plasma (FFP), almost all for TTP. There were 889 complications; 614 treatments (36% of the total) involved a complication. Most complications were minor; there were no deaths. Three treatments (0.2%) were discontinued due to a complication, and 2 (0.1%) required transfer to a higher acuity hospital bed. The most common complications were fever (7.7% of treatments), urticaria (7.4%), and hypocalcemic symptoms (7.3%). 42% of treatments with FFP involved a complication, compared to 30% of treatments using albumin-saline (P < 0.0001). The most common complications with FFP were urticaria (17%) and pruritus (13%); these occurred more commonly than in patients receiving albumin-saline. The most common complications with albumin-saline replacement were hypocalcemic symptoms (8.2%) and mild hypotension (8.1%). Mild and severe hypotension was significantly (P < 0.0001) more common with albumin-saline replacement. TPE is associated with a number of minor complications. Complications occur more commonly with FFP replacement compared to albumin-saline replacement. Pruritus and urticaria occur more commonly with FFP, and hypotension occurs more commonly with albumin-saline.


Data characterizing tensile behavior of cenosphere/HDPE syntactic foam.
The data set presented is related to the tensile behavior of cenosphere reinforced high density polyethylene syntactic foam composites "Processing of cenosphere/HDPE syntactic foams using an industrial scale polymer injection molding machine" (Bharath et al., 2016) [1]. The focus of the work is on determining the feasibility of using an industrial scale polymer injection molding (PIM) machine for fabricating syntactic foams. The fabricated syntactic foams are investigated for microstructure and tensile properties. The data presented in this article is related to optimization of the PIM process for syntactic foam manufacture, equations and procedures to develop theoretical estimates for properties of cenospheres, and microstructure of syntactic foams before and after failure. Included dataset contains values obtained from the theoretical model.


The role of sesquiterpene lactones in contact hypersensitivity to some North and South American species of feverfew (Parthenium-Compositae).
Sixteen taxa of the genus Parthenium from North and South America and 20 sesquiterpene lactones derived from plants of the Compositae family were used for patch testing six volunteers who were known to be contact sensitive to plants and lactones of this family. The patterns of cross-sensitivity and structure-activity relationships of the allergenic lactones are evaluated and the clinical and ecological relevance is discussed.


MiR-423-5p activated by E2F1 promotes neovascularization in diabetic retinopathy by targeting HIPK2.
Diabetic retinopathy (DR) is a diabetic complication and the primary cause of blindness in the world. However, the treatments of DR are challenging given its complicated pathogenesis. Here, we investigated the molecular mechanisms of DR by focusing on the function of E2F1/miR-423-5p/HIPK2/HIF1α/VEGF axis. Cultured retinal endothelial cells (hRMECs, hRECs) were treated with 25 mM glucose to mimic the high glucose-induced DR in vitro. Streptozotocin (STZ) was injected into mice to induce DR in mice. qRT-PCR, western blotting, immunohistochemistry, and ELISA were employed to measure levels of E2F1, miR-423-5p, HIPK2, HIF1α, and VEGF. H&E staining was utilized to examine retinal neovascularization. CCK-8 assay, transwell assay, and vascular tube formation assay were used to assess the cell viability, migration, and angiogenesis. Dual luciferase assay was performed to validate interactions between E2F1 and miR-423-5p, miR-423-5p and HIPK2. HG treatment increased the cell viability, migration, and angiogenesis accompanied by upregulation of E2F1, miR-423-5p, HIF1α, and VEGF levels, but reduction in HIPK2 expression. Knockdown of E2F1 or miR-423-5p suppressed the HG-induced increases in cell viability, migration, and angiogenesis. E2F1 transcriptionally activated miR-423-5p expression and miR-423-5p mimics blocked the effects of E2F1 knockdown on angiogenesis. Moreover, miR-423-5p directly targeted HIPK2 to disinhibit HIF1α/VEGF signaling. Knockdown of HIPK2 reversed the effects of miR-423-5p inhibitor on cell viability, migration, and angiogenesis. Knockdown of E2F1 suppressed neovascularization during DR in vivo. E2F1 activates miR-423-5p transcription during DR to promote angiogenesis via suppressing HIPK2 expression to disinhibit HIF1α/VEGF signaling. Strategies targeting E2F1/miR-423-5p/HIPK2 axis could be potentially used for DR treatment.


Prescribed heart failure pharmacotherapy: How closely do GPs adhere to treatment guidelines?
Heart failure patients need GPs to adhere to treatment guidelines to improve health outcomes. To examine the extent to which pharmacotherapy of heart failure patients referred for a Home Medicines Review (HMR) adheres to heart failure management guidelines. This retrospective study examined HMR reports of patients with heart failure, who were referred for a HMR due to polypharmacy. Analysis established current pharmacotherapy patients were taking and whether it aligned with heart failure guidelines. Further analysis established the types of recommendations made to the referring GPs, in order to optimise heart failure pharmacotherapy. There were 258 patients referred for HMRs between 2015 and 2018; 30 patients had a diagnosis of heart failure and therefore included in the analysis. Out of these, 13 patients were not on an ACEI and 23 were not on an AR2B; seven patients were neither on ACEI nor AR2B. The pharmacist recommended commencing ACEI in 4 patients and an AR2B in 2 patients. Nine patients were not on any B-blocker, and 4 patients were recommended to commence a B-blocker. Most patients were on suboptimal doses of their heart failure medicines, for which dose up-titrations were recommended. There was a recommendation to assess 6 patients for symptoms indicative of heart failure exacerbation, e.g. SOB. Heart failure pharmacotherapy in patients referred for HMRs was not optimised. Pharmacists play a crucial role in optimising the treatment of heart failure, and in prompting GPs to align pharmacotherapy with treatment guidelines.


In vitro microarray analysis identifies genes in acute-phase response pathways that are down-regulated in the liver of chicken embryos exposed in ovo to PFUdA.
Perfluoroundecanoic acid (PFUdA) is one of the most highly detected perfluoroalkyl compounds in wild bird tissues and eggs. Although PFUdA does not affect hatching success, many PFCs are known to impair post-hatch development and survival. Here we use microarrays to survey the transcriptional response of cultured chicken embryonic hepatocytes (CEH) to PFUdA for potential targets of PFUdA action that could lead to developmental deficiencies in exposed birds. At 1 μM and 10 μM PFUdA significantly altered the expression of 346 and 676 transcripts, respectively (fold-change>1.5, p<0.05, false discovery rate-corrected). Using functional, pathway and interactome analysis we identified several potentially important targets of PFUdA exposure, including the suppression of the acute-phase response (APR). We then measured the expression of five APR genes, fibrinogen alpha (fga), fibrinogen gamma (fgg), thrombin (f2), plasminogen (plg), and protein C (proC), in the liver of chicken embryos exposed in ovo to PFUdA. The expression of fga, f2, and proC were down-regulated in embryo livers (100 or 1000 ng/g, p<0.1) as predicted from microarray analysis, whereas fibrinogen gamma (fgg) was up-regulated and plg was not significantly affected. Our results demonstrate the utility of CEH coupled with transcriptome analysis as an in vitro screening tool for identifying novel effects of toxicant exposure. Additionally, we identified APR suppression as a potentially important and environmentally relevant target of PFUdA. These findings suggest in ovo exposure of birds to PFUdA may lead to post-hatch developmental deficiencies, such as impaired inflammatory response.


Minor methylated pyranoamentoflavones from Calophyllum venulosum.
Two new biflavonoids, pyranoamentoflavone 7-methyl ether (1) and pyranoamentoflavone 4'-methyl ether (2), have been isolated from the leaves of Calophyllum venulosum. The structures of these two new compounds were elucidated by spectroscopic data.


Genome-wide methylome pattern predictive network analysis reveal mesenchymal stem cell's propensity to undergo cardiovascular lineage.
Mesenchymal stem cells (MSCs) differentiation toward cardiovascular lineage prediction using the global methylome profile will highlight its prospective utility in regenerative medicine. We examined the propensity prediction to cardiovascular lineage using 5-Aza, a well-known cardiac lineage inducer. The customized 180 K microarray was performed and further analysis of global differentially methylated regions by Ingenuity pathway analysis (IPA) in both MSCs and 5-AC-treated MSCs. The cluster enrichment tools sorted differentially enriched genes and further annotated to construct the interactive networks. Prediction analysis revealed pathways pertaining to the cardiovascular lineage found active in the native MSCs, suggesting its higher propensity to undergo cardiac, smooth muscle cell, and endothelial lineages in vitro. Interestingly, gene interaction network also proposed majorly stemness gene network NANOG and KLF6, cardiac-specific transcription factors GATA4, NKX2.5, and TBX5 were upregulated in the native MSCs. Furthermore, the expression of cardiovascular lineage specific markers such as Brachury, CD105, CD90, CD31, KDR and various forms of ACTIN (cardiac, sarcomeric, smooth muscle) were validated in native MSCs using real time PCR and immunostaining and blotting analysis. In 5-AC-treated MSCs, mosaic interactive networks were observed to persuade towards osteogenesis and cardiac lineage, indicating that 5-AC treatment resulted in nonspecific lineage induction in MSCs, while MSCs by default have a higher propensity to undergo cardiovascular lineage. The online version contains supplementary material available at 10.1007/s13205-021-03058-2.


Far infrared (terahertz) spectroscopy of a series of polycyclic aromatic hydrocarbons and application to structure interpretation of asphaltenes and related compounds.
A series of 33 different polycyclic aromatic hydrocarbons (PAHs) were studied by far infrared spectroscopy (terahertz spectroscopy) in the spectral range comprised between 600 and 50 cm(-1). In addition to common PAHs like naphthalene, anthracene, phenanthrene, fluoranthene, picene, pyrene, benzo[α]pyrene, and perylene, also quite unusual PAHs were studied like tetracene, pentacene, acenaphtene, acenaphtylene, triphenylene, and decacyclene. A series of alkylated naphthalenes and anthracenes were studied as well as methypyrene. Partially or totally hydrogenated PAHs were also object of the present investigation, ranging from tetrahydronaphthalene (tetralin) to decahydronaphthalene (decalin), 9,10-dihydroanthracene, 9,10-dihydrophenanthrene, hexahydropyrene, and dodecahydrotriphenylene. Finally, the large and quite rare PAHs coronene, quaterrylene, hexabenzocoronene, and dicoronylene were studied by far infrared spectroscopy. The resulting reference spectra were used in the interpretation of the chemical structure of asphaltenes (as extracted from a heavy petroleum fraction and from bitumen), the chemical structures of other petroleum fractions known as DAE (distillate aromatic extract) and RAE (residual aromatic extract), and a possible interpretation of components of the chemical structure of anthracite coal. Asphaltenes, heavy petroleum fractions, and coal were proposed as model compounds for the interpretation of the emission spectra of certain proto-planetary nebulae (PPNe) with a good matching in the mid infrared between the band pattern of the PPNe emission spectra and the spectra of these oil fractions or coal. Although this study was finalized in an astrochemical context, it may find application also in the petroleum and coal chemistry.


Effects of a brine discharge over soft bottom Polychaeta assemblage.
Desalination is a growing activity that has introduced a new impact, brine discharge, which may affect benthic communities. Although the role of polychaetes as indicators to assess organic pollution is well known, their tolerance to salinity changes has not been examined to such a great extent. The aim of this study was to examine the effect of brine discharge over soft bottom polychaete assemblage along the Alicante coast (Southeast Spain) over a two year period. Changes in the polychaete assemblage was analysed using univariate and multivariate techniques. We compared a transect in front of the discharge with two controls. At each transect we sampled at three depths (4, 10 and 15 m) during winter and summer. We have observed different sensitivity of polychaete families to brine discharges, Ampharetidae being the most sensitive, followed by Nephtyidae and Spionidae. Syllidae and Capitellidae showed some resistance initially, while Paraonidae proved to be a tolerant family.


Feasibility of six-month outpatient cognitive remediation in schizophrenia: Experience from the randomized controlled integrated social cognition and social skills therapy study.
Patients with schizophrenia often have cognitive impairments that contribute to diminished psychosocial functioning. Cognitive remediation therapy (CRT) has proven efficacy and is recommended by evidence-based treatment guidelines. Important moderators of efficacy include integration of CRT into a psychiatric rehabilitation concept and patient attendance at a sufficient number of therapy sessions. These conditions can probably best be met in an outpatient setting; however, outpatient treatment is prone to higher rates of treatment discontinuation and outpatient settings are not as well protected as inpatient ones and less closely supervised.The present study investigated the feasibility of outpatient CRT in schizophrenia over a six-month period. Adherence to scheduled sessions and safety parameters were assessed in 177 patients with schizophrenia randomly assigned to one of two matched CRT programs.Results showed that 58.8 % of participants completed the CRT (>80 % of scheduled sessions) and 72.9 % completed at least half the sessions. Predictor analysis revealed a high verbal intelligence quotient as favorable for good adherence, but this factor had only low general predictive power. During the six-month treatment phase, serious adverse events occurred in 15.8 % (28/177) of the patients, which is a comparable rate to that reported in the literature.Our findings support the feasibility of six-month outpatient CRT in schizophrenia in terms of adherence to scheduled sessions and safety. NCT02678858, DRKS00010033.


Failure to return pillbox is a predictor of being lost to follow-up among people living with HIV on antiretroviral therapy in rural Tanzania.
Pill count is used to assess drug adherence in people living with HIV (PLHIV). Carrying a pillbox is associated with fear of concealment and stigma and might indicate poor adherence and predict someone who will be lost to follow-up (LTFU). We therefore assessed the association between pillbox return and being LTFU in rural Tanzania. This is a nested study of the Kilombero and Ulanga Antiretroviral Cohort (KIULARCO). We included PLHIV aged ≥ 18 years enrolled in KIULARCO between January 2013 and March 2019 with follow-up through January 2020, who were on antiretroviral treatment (ART) for ≥ 6 months. Baseline was defined as the latest ART initiation or KIULARCO enrolment. We determined the association between time-dependent failed pillbox return updated at every visit and LTFU using Kaplan-Meier estimation and Cox models. Among 2552 PLHIV included in the study, 1735 (68.0%) were female, 959 (40.3%) had a WHO stage III/IV and 1487 (66.4%) had a CD4 cell count < 350 cells/µL. The median age was 38.4 years [interquartile range (IQR): 31.7-46.2]. During a median follow-up of 33.1 months (IQR: 17.5-52.4), 909 (35.6%) participants were LTFU, 43 (1.7%) died and 194 (7.6%) had transferred to another clinic. The probability of being LTFU was higher among PLHIV with failed pillbox return than among those who returned their pillbox [30.0%, 95% confidence interval (CI): 26.8-33.2% vs. 19.4%, 95% CI: 17.4-21.6%, respectively, at 24 months (hazard ratio = 1.67, 95% CI: 1.46-1.90; p < 0.001)]. Failed pillbox return was associated with a higher risk of being LTFU and could be used as a simple tool to identify PLHIV for appropriate interventions to reduce their chance of being LTFU.


IL-10 production in circulating T cells differs between allergen-induced isolated early and dual asthmatic responders.
IL-10 is an anti-inflammatory cytokine released from various cells, including T cells. The role of IL-10 in asthma pathogenesis remains uncertain. Allergen inhalation by atopic asthmatic subjects results in 2 bronchoconstrictor phenotypes: isolated early response and dual response. Persistence of allergen-induced airway inflammation is a feature of dual responders. The kinetics of IL-10 production in circulating T cells were investigated to examine a potential role of IL-10 in allergen-induced responses and airway inflammation. Fourteen subjects with mild asthma (7 isolated early and 7 dual responders) were challenged with allergen. PBMCs taken before and 24 hours after allergen challenge were processed for intracellular IL-10 staining with fluorescent-conjugated anti-IL-10 antibody. The frequency of IL-10-producing cells was assessed for CD4(+) and CD8(+) T-cell subsets by using flow cytometry. Before allergen administration, the frequency of IL-10-producing CD4(+) cells was significantly higher in dual than in isolated early responders. IL-10-producing CD4(+) cells significantly increased after allergen in early responders, whereas IL-10-producing CD4(+) cells significantly decreased in dual responders. Simultaneous assessments of IL-5-producing T cells did not show any differences between each group before or after allergen administration. These results suggest that the contrasting profiles of IL-10 production may be associated with the different time course of allergen-induced airway inflammation between allergen-induced early and dual responders.


Aesthetic Facial Surgery and Orthodontics: Common Goals.
Many of the aesthetic facial procedures can be performed simultaneously at the time of initial orthognathic surgery. Correction of any residual deformities after surgery, such as mandibular notching, malar asymmetry, labiomental crease, and any camouflage treatment, should be performed as a delayed procedure, when the outcome is more predictable. Additionally, these procedures could be used to enhance the orthodontic result, without the need of osteotomies to reposition the bones.


Sustained release of arsenic trioxide benefits interventional therapy on rabbit VX2 liver tumor.
The benefit of chemotherapy as a constituent of transcatheter arterial chemoembolization (TACE) is still in debate. Recently we have developed arsenic trioxide nanoparticle prodrug (ATONP) as a new anticancer drug, but its systemic toxicity is a big issue. In this preclinical TACE study, ATONP emulsified in lipiodol behaved as drug-eluting bead manner. Sustained release of arsenic from ATONP within occluded tumor caused very low arsenic level in plasma, avoiding the "rushing out" effect as ATO did. Correspondingly, intratumoral arsenic accumulation and inorganic phosphate deprivation were simultaneously observed, and arsenic concentration was much higher as ATONP was transarterially administered than ATO, or intravenously injected. Tumor necrosis and apoptosis were remarkably more severe in ATONP group than ATO, but no significant hepatic and renal toxicity was perceived. In brief, ATONP alleviated arsenic toxicity and boosted the therapeutic effect of TACE via Pi-activated drug sustainable release.


Developing Aspergillus niger as a cell factory for food enzyme production.
Aspergillus niger has become one of the most important hosts for food enzyme production due to its unique food safety characteristics and excellent protein secretion systems. A series of food enzymes such as glucoamylase have been commercially produced by A. niger strains, making this species a suitable platform for the engineered of strains with improved enzyme production. However, difficulties in genetic manipulations and shortage of expression strategies limit the progress in this regard. Moreover, several mycotoxins have recently been detected in some A. niger strains, which raises the necessity for a regulatory approval process for food enzyme production. With robust strains, processing engineering strategies are also needed for producing the enzymes on a large scale, which is also challenging for A. niger, since its culture is aerobic, and non-Newtonian fluid properties are developed during submerged culture, making mixing and aeration very energy-intensive. In this article, the progress and challenges of developing A. niger for the production of food enzymes are reviewed, including its genetic manipulations, strategies for more efficient production of food enzymes, and elimination of mycotoxins for product safety.


Source identification, contamination status and health risk assessment of heavy metals from road dusts in Dhaka, Bangladesh.
In this study, concentrations of Cr, Mn, Ni, Cu, Zn, Cd and Pb were determined in road dusts collected from different locations in Dhaka to assess source, contamination status and health risk. Energy-dispersive X-ray fluorescence spectroscopy and energy-dispersive X-ray spectroscopy were used to determine Cr, Mn, Ni, Cu, Zn, Cd and Pb and their mean concentrations were 162.27 ± 29.46, 721.18 ± 180.14, 35.65 ± 12.55, 104.56 ± 128.33, 515.32 ± 321.90, BDL, and 342.82 ± 591.20 mg/kg, respectively. Among the heavy metals, highest concentrations of Cu, Zn and Pb were found at urban sites-7 (municipal waste dumping) and 8 (medical waste incineration). Highest concentration of Cr followed by Cu and Zn was found at site-5 (Tejgaon, urban). Principal component analysis revealed that anthropogenic activities are the potential sources for Cr, Ni, Cu, Zn and Pb while earth crust for Mn. Pollution index and pollution load index results suggested that all the sites were contaminated and/or degraded by Cr, Cu, Zn and Pb except sites-9 (urban), 10 (sub-urban), 11 (rural) while sites-7 and 8 (urban) were extremely degraded. For noncarcinogenic health risk, hazard quotient values for dermal were higher compared to that of inhalation/ingestion. Though hazard index values were less than 1 at all the sites, these were at least one order of magnitude higher for children group than that of adult group, thus the children group may face more noncarcinogenic health risk at sites-7 and 8. Values of incremental lifetime cancer risk were from 10-9 to 10-11 showed no carcinogenic health risk by road dusts contaminated with the heavy metals.


Electron microscopy of serotonin-immunoreactive neuron branches and terminals in the locust central nervous system.
Using a pre-embedding peroxidase technique, we have investigated the ultrastructure of elements that react with antibodies to 5-hydroxytryptamine in the central nervous system of the locust. Reactive neuron profiles are widespread, and contain a variety of vesicle types: small lucent vesicles, some of which are stained only on their outer membranes while others are stained internally, and large dense granules that again differ in the staining properties of their cores. Some of the reactive profiles contain synaptic specializations, while others receive synaptic inputs from unlabelled elements. The heterogeneity of reactive profiles makes it impossible to define ultrastructural characteristics of putative serotonergic terminals that might be generally applicable. The differential reactivity both of small vesicles and of large dense granules may indicate functional differences within these categories.


PICOT, protein kinase C theta-interacting protein, is a novel regulator of FcepsilonRI-mediated mast cell activation.
PICOT (PKC-interacting cousin of thioredoxin) consists of one thioredoxin homology domain in the N-terminal and two tandem PICOT homology domains in the C-terminal. PICOT specifically interacts with protein kinase C theta (PKC-theta) via its thioredoxin homology domain and acts as an important modulator of T cell receptor (TCR)-signaling. Using PICOT overexpressing rat basophilic leukemia cells (RBL-2H3), we evaluated the effect of PICOT overexpression on the FcepsilonRI-mediated signaling. In comparison to the control cells, introduction of PICOT to RBL-2H3 cells induced increased degranulation and the activation of NFAT and in the expression of IL-4 and TNF-alpha transcripts by FcepsilonRI-crosslinking, whereas no significant change was observed with the elevation of ERK1/2 and p38 MAP kinase phosphorylation and NF-kappaB activation by FcepsilonRI aggregation. More interesting was the exogenous PICOT overexpression in RBL-2H3 cells causing a large decrease in the elevation of JNK phosphorylation. PICOT-regulated FcepsilonRI-mediated signals in RBL-2H3 cells and acted as a positive regulator on IL-4 and TNF-alpha expression, NFAT and degranulation signal pathways and a negative regulator on a JNK signal pathway. Considering that PICOT has no enzymatic activity, the regulation of PICOT on FcepsilonRI-signaling may depend on PICOT-associated molecule(s).


E. coli DNA binding protein HU forms nucleosomelike structure with circular double-stranded DNA.
The incubation of the E coli DNA binding protein HU with relaxed circular SV40 DNA in the presence of pure nicking-closing enzyme introduces up to 18 negative superhelical turns in the DNA molecules as measured by agarose gel electrophoresis. The maximal density of supercoiling is obtained at a HU-DNA mass ratio of 1. Reconstituted DNA-HU complexes prefixed with glutaraldehyde appear as condensed circular structures having an average of 14 "beads" per circular SV40 DNA molecule, with a "bead" diameter of 180 +/- 23 A. The circular SV40 DNA is condensed by a ratio of 2.0-2.5 relative to naked DNA. This is similar to the ratio (2.4) measured for chromatin formed by reassociation of relaxed SV40 DNA with the four core histones.


SC3 monoclonal antibody defines a novel specific human B-cell surface antigen differentially expressed on B-cell leukaemias and lymphomas and involved in the proliferation of normal and malignant B lymphocytes.
The monoclonal antibody SC3 was raised against the NK leukaemia cell line YTindi. It detected a 98-kDa surface antigen with weak expression on a restricted number of leukaemia cell lines under reducing conditions. SC3 mAb labelled 5-10% of normal peripheral blood lymphocytes corresponding almost exclusively to B lymphocytes, and 60-70% of tonsillar B cells. It did not react with erythrocytes, platelets or monocytes whereas it stained granulocytes. The aim of the present study was to examine the expression and functional effects of SC3 mAb reactive epitope on normal and malignant B cells. Most SC3+ B cells from healthy donors were CD23+, some co-expressed CD5 and CD27 and a few were CD38+. SC3 epitope was expressed exclusively by B-lineage malignant proliferations, including B-lineage ALL. Practically, all B-CLL studied expressed SC3 mAb reactive epitope although with variable intensity, while MCL and PLL were negative. Other low grade and high grade B-NHL were variably stained. SC3 mAb alone triggered the proliferation of CD2-depleted PBL and significantly increased the proliferation induced by suboptimal concentrations of LPS. This effect was much weaker with B-CLL cells but was increased after cross-linking with an anti-IgM antibody. The restricted expression pattern combined with molecular weight and functional data indicate that SC3 mAb may detect a novel B-cell antigen mostly expressed by early and naive B cells. Although its expression in B-cell malignancies was not limited to a single differentiation stage, it might confer specific functional characteristics to the positive malignant cells.


Aseasonal leaf abscission in Populus induced by volcanic ash.
Premature leaf abscission in Populus was observed in central Washington (USA) about two weeks after the 18 May 1980 ashfall from Mount St. Helens. Leaf abscission was probably a wounding response caused by sandsized ash carried in winds that gusted to 15 m s-1 7 days after the ashfall. Under glasshouse conditions simulating this environmental regime the leaf epidermis and often part of the mesophyll in Populus nigra L. var. italica Duroi. were abraded, resulting in permanent loss of leaf turgor and decreased stomatal conductance. Substantial abscission of still-green leaves ensued. Leaf abrasion by ash-laden winds, although rare, may result in conspicuous plant damage.


[Automatic dosage of uric acid by urate oxydase. Comparison with Praetorius' methods].
The authors describe an automatic method of estimation of uric acid based on the colour obtained in the presence of Gaiac resin and peroxidase, and the hydrogen peroxide produced by the action of urate oxidase on uric acid.


Revisiting lymphocyte clonality testing in feline B-cell lymphoma.
Differentiation between resident mature lymphocyte populations and small-cell lymphoma cannot be made by cytological review alone and remains challenging in histopathological review. These cases warrant application of complementary tools like PCR-based immunoglobulin (IG) and T-cell receptor (TCR) clonality testing for confirmation. Although primer coverage of potential IG gene rearrangements in feline B-cell neoplasms constantly improves, the possibility of false negative and false positive test results still poses a problem. In this retrospective study, we assessed diagnostic sensitivity and specificity of a novel developed multiplex PCR assay for routine diagnosis of B-cell clonality. Therefore, 24 feline patients were subjected to comparative clonality testing by using different primer sets. Feline lymphoma cell lines and confirmed patient material served as positive control. Compared to previous studies, this novel developed multiplex PCR assay showed positive effects on diagnostic sensitivity, specificity, accuracy, and positive predictive value accompanied by a slight impairment of negative predictive value. Notably, none of the primer sets was superior; hence, we recommend the combined application of the herein tested primer sets in routine diagnostics. However, a more in-depth-evaluation of the dynamic of assay specific parameters in dependency on primer set usage requires prospective studies on larger cohorts of feline patients.


Repetitive contractions of the upper esophageal body and sphincter in achalasia.
This report describes repetitive contractions in the upper esophageal sphincter (UES) and the repetitive upper esophageal spontaneous contractions (RUESCs) of patients with achalasia and relates this activity to repetitive contractile activity (RCA) recorded in the more distal esophageal body, to intraesophageal pressure (IEP), and to lower esophageal sphincter (LES) pressure. Two hundred and sixteen consecutive esophageal motility studies from 156 achalasia patients with retrospectively assessed. RUESCs were found in 105 patients (67%) and 125 of 216 studies (58%). General features of the RUESC were (1) coincidence with simultaneous repetitive increases in pressure throughout the entire esophageal body; (2) amplitude of pressure increases tended to be higher in the proximal esophagus; (3) RUESC frequency was different than respiration, except for 6 cases where continuous, RUESC and RCA were synchronized with inspiration; and (4) RUESCs were positively associated with increased IEP, and with increased LES pressure (> 40 mmHg). RCA in the esophageal body was uncommon without RUESC. It is concluded that (1) RUESCs are common in achalasia and appear to be closely linked to contractile activity in the upper esophageal body; (2) the close relationship of RUESC and RCA in the esophageal body to increased IEP and elevated LES pressure suggests that esophageal tone is high in these subjects; and (3) these findings indicate a potential mechanisms for localization of some of the clinical symptoms to the retrosternal and suprasternal areas, for the inability to readily belch, and for the development of structural features such as a prominent cricopharyngeal bar.


Heyneanol A induces apoptosis via cytochrome c release and caspase activation in human leukemic U937 cells.
Heyneanol A, a tetramer of resveratrol, is isolated from the roots of Vitis amurensis by cytotoxicity based fractionation. In this study, the mechanism of apoptosis by heyneanol A was evaluated in human leukemic U937 cells. Heyneanol A (IC(50) = 6.6 microM at 24 h) exhibited stronger cytotoxic effect than resveratrol (IC(50) = 100 microM at 24 h) by 15-fold on human leukemic U937 cells by XTT assay. Apoptotic bodies were observed in U937 cells treated with 6 microM of heyneanol A by TUNEL assay. Heyneanol A effectively increased the portion of sub-G(1) DNA content in a time- and concentration-dependent manner by flow cytometric analysis. Heyneanol A also induced cytochrome c release from mitochondria into the cytosol and subsequent caspase activation involving caspase 9 and 3 to cleave PARP. However, it did not affect the expressions of Bax and Bcl-2 by western blotting. It was confirmed that the activation of caspase 8, 9 and 3 and the cleavage of PARP by heyneanol A were completely blocked by adding Z-VAD-FMK, a caspase inhibitor. These findings suggest that heyneanol A has anti-tumor activity, which may be mediated by apoptosis caused by cytochrome c release and caspase activation in human leukemic U937 cells.