A bovine ephemeral fever vaccine incorporating adjuvant Quil A: a comparative study using adjuvants Quil A, aluminium hydroxide gel and dextran sulphate.
Various vaccines containing the 919 strain of ephemeral fever virus were evaluated in experimental calves and in commercial cattle. The vaccine virus was mixed with one of the adjuvants, Quil A (a saponin derivative), aluminium hydroxide gel, dextran sulphate or combinations of these. The response of experimental calves was evaluated by measuring the production of neutralising antibodies and by resistance to challenge with virulent virus; the response of commercial cattle was judged only by the production of neutralising antibody. Twelve calves given two doses of vaccine containing Quil A produced neutralising antibodies to bovine ephemeral fever virus and all were resistant to challenge with virulent virus given 28 to 76 days after the second vaccination. The vaccine given in three of these calves also contained aluminium hydroxide gel. Six of eight unvaccinated control calves succumbed to experimental challenge. In commercial cattle (17 to 26 animals per group) the serological response after two doses of vaccine containing Quil A or Quil A and dextran sulphate was significantly better than that after vaccines containing only dextran sulphate or after vaccines containing combinations of aluminium hydroxide gel and Quil A. The adjuvant Quil A alone was tested in cattle and shown to produce a transient soft swelling at the injection site as well as a rise in rectal temperature of greater than 1 degree C one day after inoculation. At least 99.99 per cent of viral infectivity was destroyed when the vaccine was mixed with Quil A, suggesting that live virus may not be essential in the immunogenicity of the vaccine. This vaccine overcame two of the problems associated with previous attenuated vaccines tested in Australia; the necessity for adjuvant and virus to be mixed immediately before use and the large volume of the vaccine.


Lactate uptake contributes to the NAD(P)H biphasic response and tissue oxygen response during synaptic stimulation in area CA1 of rat hippocampal slices.
Synaptic train stimulation (10 Hz x 25 s) in hippocampal slices results in a biphasic response of NAD(P)H fluorescence indicating a transient oxidation followed by a prolonged reduction. The response is accompanied by a transient tissue PO(2) decrease indicating enhanced oxygen utilization. The activation of mitochondrial metabolism and/or glycolysis may contribute to the secondary NAD(P)H peak. We investigated whether extracellular lactate uptake via monocarboxylate transporters (MCTs) contributes to the generation of the NAD(P)H response during neuronal activation. We measured the effect of lactate uptake inhibition [using the MCT inhibitor alpha-cyano-4-hydroxycinnamate (4-CIN)] on the NAD(P)H biphasic response, tissue PO(2) response, and field excitatory post-synaptic potential in hippocampal slices during synaptic stimulation in area CA1 (stratum radiatum). The application of 4-CIN (150-250 micromol/L) significantly decreased the reduction phase of the NAD(P)H response. When slices were supplemented with 20 mmol/L lactate in 150-250 micromol/L 4-CIN, the secondary NAD(P)H peak was restored; whereas 20 mmol/L pyruvate supplementation did not produce a recovery. Similarly, the tissue PO(2) response was decreased by MCT inhibition; 20 mmol/L lactate restored this response to control levels at all 4-CIN concentrations. These results indicate that lactate uptake via MCTs contributes significantly to energy metabolism in brain tissue and to the generation of the delayed NAD(P)H peak after synaptic stimulation.


Sulindac Modulates the Response of Proficient MMR Colorectal Cancer to Anti-PD-L1 Immunotherapy.
Immune-checkpoint inhibitor (ICI) therapy has been widely used to treat different human cancers, particularly advanced solid tumors. However, clinical studies have reported that ICI immunotherapy benefits only ∼15% of patients with colorectal cancer, specifically those with tumors characterized by microsatellite instability (MSI), a molecular marker of defective DNA mismatch repair (dMMR). For the majority of patients with colorectal cancer who carry proficient MMR (pMMR), ICIs have shown little clinical benefit. In this study, we examined the efficacy of sulindac to enhance the response of pMMR colorectal cancer to anti-PD-L1 immunotherapy. We utilized a CT26 syngeneic mouse tumor model to compare the inhibitory effects of PD-L1 antibody (Ab), sulindac, and their combination on pMMR colorectal cancer tumor growth. We found that mice treated with combination therapy showed a significant reduction in tumor volume, along with increased infiltration of CD8+ T lymphocytes in the tumor tissues. We also demonstrated that sulindac could downregulate PD-L1 by blocking NF-κB signaling, which in turn led to a decrease in exosomal PD-L1. Notably, PD-L1 Ab can be bound and consumed by exosomal PD-L1 in the blood circulation. Therefore, in combination therapy, sulindac downregulating PD-L1 leads to increased availability of PD-L1 Ab, which potentially improves the overall efficacy of anti-PD-L1 therapy. We also show that low-dose sulindac does not appear to have a systemic inhibitory effect on prostaglandin E2 (PGE2). In conclusion, our findings provide unique insights into the mechanism of action and efficacy for sulindac as an immunomodulatory agent in combination with anti-PD-L1 therapy for the treatment of pMMR colorectal cancer.


In search of benzene-like Sn6(6-): synthesis of Na4CaSn6 with interconnected cyclohexane-like Sn6(6-).
The title compound was synthesized in an attempt to produce stacked benzene-like Sn6(6-) rings separated by alkaline-earth cations in analogy with the recently reported stacks of aromatic cyclopentadienyl-like Sn5(6-) in Na8BaSn6 (in addition to isolated Sn4- anions). The resulting compound, synthesized from a stoichiometric mixture of the elements at high temperature, has the "correct" stoichiometry with six tin atoms and six positive charges. However, the rings of Sn6(6-) are puckered into chair-type cyclohexanes that are interconnected into isolated cylindrical tubes stuffed with Ca2+ between the rings. Such tubes, if fused to each other, form the hexagonal diamond structure. The new compound is electronically balanced according to magnetic and four-probe resistivity measurements. Reported are also the synthesis and properties of Na10EuSn12 and Na10YbSn12 which are isostructural with the known Na10CaSn12.


Prevalence and Determinants of Vaccine Hesitancy and Vaccines Recommendation Discrepancies among General Practitioners in French-Speaking Parts of Belgium.
General practitioners (GPs) play a critical role in patient acceptance of vaccination. Vaccine hesitancy (VH) is a growing phenomenon in the general population but also affects GPs. Few data exist on VH among GPs. The objectives of this analysis of a population of GPs in the Belgian Wallonia-Brussels Federation (WBF) were to: (1) determine the prevalence and the features of VH, (2) identify the correlates, and (3) estimate the discrepancy in vaccination's behaviors between the GPs' children and the recommendations made to their patients. An online survey was carried out among the population of general practitioners practicing in the WBF between 7 January and 18 March 2020. A hierarchical cluster analysis was carried out based on various dimensions of vaccine hesitancy: perception of the risks and the usefulness of vaccines as well as vaccine recommendations for their patients. A total of 251 GPs answered the survey. The average percentage of moderate to high vaccine hesitancy was 50.6%. Three factors were independently associated with increased risk of vaccine hesitancy: an age <50 years old, having no children, and having no contact with selected vaccine-preventable disease (measles, complicated influenza, chronic hepatitis B (HBV), bacterial meningitis, or cervical cancer) in the past 5 years. VH was associated with controversies on vaccines' safety. GPs who had vaccinated their children against six diseases (MMR, meningococcus C (MenC), HBV, and HPV) tended not to recommend the same vaccines to their patients. Among GPs with all children vaccinated against HBV, only 37.5% recommended catch-up HBV immunization to their patients. In this small cohort of GP, moderate to high VH was associated with controversies on vaccines' safety and with specific personal characteristics (age <50, no children, and no recent experience with a serious VPD). As previously reported, GPs have different vaccine prescription attitude toward their patients and children. These findings should be confirmed in larger cohorts.


Identifying latent classes of empowerment among early adolescent girls and the factors that cultivate them: Evidence from a community-based skills training program.
Efforts to empower adolescent girls remain constrained by confusion around how to conceptually define empowerment and how to design interventions. More research, particularly among young adolescents, is needed that considers contextual and age-specific circumstances. This paper examines the latent classes of empowerment among early adolescent girls in Bangladesh and factors associated with membership in these classes. Using data from the Bangladeshi Association for Life Skills, Income, and Knowledge for Adolescents project, a four-arm randomized controlled trial designed to evaluate different community-based approaches for adolescents' skills development, we identify the latent classes before and after intervention and explore whether girls' membership in the study arms was associated with latent class membership at endline. To identify underlying subgroups among the sample, we conducted latent class analysis at baseline and endline using indicators of seven different constructs of empowerment based on the literature. Using multinomial logistic regression analysis, we assessed whether respondents' background characteristics and membership in the study arms were predictive of latent class membership at both timepoints. A three-class model at baseline and a four-class model at endline showed optimal fit. In both models, three distinct latent classes were observed: (1) a least empowered class, (2) a mobile, socially active class, and (3) a socially progressive class. At endline, a fourth class emerged-the most empowered-that was mobile, socially active, and socially progressive. Girls' marital status, religion, and parents' level of education predicted latent class membership; household wealth was predictive at baseline, but not endline. Study arm was also associated with class membership at endline: girls in the Gender-Rights Awareness Arm had the highest probability of belonging to the fourth, most empowered class. This paper identifies and characterizes latent classes of empowerment that exist among the sample before and after the intervention. The apparent differential effects of interventions in predicting latent class membership have implications for community-based programming. Findings suggest that skills-building curriculum focused on promoting gender equitable norms, voice, and agency has potential to influence early adolescent girls across multiple dimensions of empowerment.


Pulmonary toxicity from novel antineoplastic agents.
The pulmonary side-effects induced by novel antineoplastic agents have not been well characterized. To further investigate this topic, relevant English and non-English language studies were identified through Medline. For our search we used the generic names of novel cytotoxic or non-cytotoxic antineoplastic agents and the key phrases pulmonary/lung toxicity, dyspnea, pneumonitis, acute lung injury, acute respiratory distress syndrome and alveolar damage. The references from the articles identified were reviewed for additional sources. Abstracts from International Meetings were also included. Furthermore, information was obtained from the Pneumotox website, which provides updated knowledge on drug-induced respiratory disease as well as from pharmaceutical websites. Most novel antineoplastic drugs may induce pulmonary toxicity, which involves mainly the parenchyma, and less frequently the airways, pleura or the pulmonary circulation. Furthermore, a subset of these agents impairs pulmonary function tests. The exact incidence of lung toxicity remains unclear. The most common patterns consist of dyspnea without further details and infiltrative lung disease (ILD), denoting changes in the interstitium or alveoli. The diagnosis is one of exclusion. ILD is usually benign and responds to appropriate treatment; however, fatalities have been reported. Clinicians should be aware of the potential of most novel antineoplastic agents to cause lung toxicity. A high index of suspicion is required if these are combined with other cytotoxic drugs or radiation.


Deep learning resting state functional magnetic resonance imaging lateralization of temporal lobe epilepsy.
Localization of focal epilepsy is critical for surgical treatment of refractory seizures. There remains a great need for noninvasive techniques to localize seizures for surgical decision-making. We investigate the use of deep learning using resting state functional magnetic resonance imaging (RS-fMRI) to identify the hemisphere of seizure onset in temporal lobe epilepsy (TLE) patients. A total of 2132 healthy controls and 32 preoperative TLE patients were studied. All participants underwent structural MRI and RS-fMRI. Healthy control data were used to generate training samples for a three-dimensional convolutional neural network (3DCNN). RS-fMRI was synthetically altered in randomly lateralized regions in the healthy control participants. The model was then trained to classify the hemisphere containing synthetic noise. Finally, the model was tested on TLE patients to assess its performance for detecting biological seizure onset zones, and gradient-weighted class activation mapping (Grad-CAM) identified the strongest predictive regions. The 3DCNN classified healthy control hemispheres known to contain synthetic noise with 96% accuracy, and TLE hemispheres clinically identified to be seizure onset zones with 90.6% accuracy. Grad-CAM identified a range of temporal, frontal, parietal, and subcortical regions that were strong anatomical predictors of the seizure onset zone, and the resting state networks that colocalized with Grad-CAM results included default mode, medial temporal, and dorsal attention networks. Lastly, in an analysis of a subset of patients with postsurgical outcomes, the 3DCNN leveraged a more focal set of regions to achieve classification in patients with Engel Class >I compared to Engel Class I. Noninvasive techniques capable of localizing the seizure onset zone could improve presurgical planning in patients with intractable epilepsy. We have demonstrated the ability of deep learning to identify the correct hemisphere of the seizure onset zone in TLE patients using RS-fMRI with high accuracy. This approach represents a novel technique of seizure lateralization that could improve preoperative surgical planning.


Antihypertensive effect of celery seed on rat blood pressure in chronic administration.
This study investigated the effects of different celery (Apium graveolens) seed extracts on blood pressure (BP) in normotensive and deoxycorticosterone acetate-induced hypertensive rats. The hexanic, methanolic, and aqueous-ethanolic extracts were administered intraperitoneally and their effects on BP and heart rate (HR) were evaluated in comparison with spirnolactone as a diuretic and positive control. Also, the amount of n-butylphthalide (NBP), as an antihypertensive constituent, in each extract was determined by HPLC. The results indicated that all extracts decreased BP and increased the HR in hypertensive rats, but had no effect on normotensive rats. The data showed that administration of 300 mg/kg of hexanic, methanolic, and aqueous-ethanolic (20/80, v/v) extracts of the celery seed caused 38, 24, and 23 mmHg reduction in BP and 60, 25, and 27 beats per minute increase in the HR, respectively. Also, the HPLC analysis data revealed that the content of NBP in the hexanic extract was 3.7 and 4 times greater than methanolic and aqueous-ethanolic extracts. It can be concluded that celery seed extracts have antihypertensive properties, which appears to be attributable to the actions of its active hydrophobic constitutes such as NBP and can be considered as an antihypertensive agent in chronic treatment of elevated BP.


Calcium absorption in corticoid treated subjects effects of a single oral dose of calcitriol.
We compared the fractional absorption of calcium (FACa, 6 h, % TD) and the radiocalcium transit (% TD per min) in seven glucocorticoid-treated patients (10-25 mg prednisolone per day) and in seven normal subjects, in the basal state and 12 h after an oral dose of synthetic 1,25-(OH)2D (3 micrograms). In the basal state, the radiocalcium transit was significantly decreased (P less than 0.02) at 15 min in patients treated with prednisolone, but FACa at 6 h was not significantly decreased (51 +/- 5 vs. 60 +/- 5% TD). 12 h after an oral dose of 1,25-(OH)2D which resulted in supraphysiologic plasma levels, FACa increased significantly (P less than 0.02) in both groups but the peak absorption rate of Ca remained lower in the corticoid-treated patients than in controls (P less than 0.02). The results suggest that glucocorticoids decrease the 1,25-(OH)2D-dependent transport of calcium across the proximal small intestine.


beta-D-glucuronidase gene from Escherichia coli is a functional reporter in the methylotrophic yeast Pichia pastoris.
The Escherichia coli gene gusA was expressed in the methylotrophic yeast Pichia pastoris in a transcriptional fusion to the homologous methanol-inducible AOX1 promoter. Four recombinant clones were selected for expression studies in shake flask conditions and beta-D-glucuronidase (beta-GUS) activity was assayed each 24 h during the induction period. Regardless of the genomic integration patterns and the gene dosage, beta-GUS was functionally expressed and easily detected in all studied clones. The results obtained demonstrate the feasibility of using this bacterial enzyme as a reporter in Pichia pastoris.


[The characteristics of aerophagy and its treatment].
Clinical and roentgenological examination of esophagus and cervical spine were carried out in 27 patients with functional aerophagia. In 11 of them the air was found to pass through without any swallowing into the stomach and in remaining 16--into esophagus. All the patients revealed osteochondrosis, unstable and blocked spines. Manual therapy applied to cervical zone of spine promoted elimination of aerophagia.


Childhood Disadvantage Moderates Late Midlife Default Mode Network Cortical Microstructure and Visual Memory Association.
Childhood disadvantage is a prominent risk factor for cognitive and brain aging. Childhood disadvantage is associated with poorer episodic memory in late midlife and functional and structural brain abnormalities in the default mode network (DMN). Although age-related changes in DMN are associated with episodic memory declines in older adults, it remains unclear if childhood disadvantage has an enduring impact on this later life brain-cognition relationship earlier in the aging process. Here, within the DMN, we examined whether its cortical microstructural integrity-an early marker of structural vulnerability which increases risk for future cognitive decline and neurodegeneration-is associated with episodic memory in adults at ages 56-66, and whether childhood disadvantage moderates this association. Cortical mean diffusivity (MD) obtained from diffusion magnetic resonance imaging was used to measure microstructural integrity in 350 community-dwelling men. We examined both visual and verbal episodic memory in relation to DMN MD and divided participants into disadvantaged and non-disadvantaged groups based on parental education and occupation. Higher DMN MD was associated with poorer visual memory but not verbal memory (β=-.11, p=.040 vs. β=-.04, p=.535). This association was moderated by childhood disadvantage and was significant only in the disadvantaged group (β=-.26, p=.002 vs. β=-.00, p=.957). Lower DMN cortical microstructural integrity may reflect visual memory vulnerability in cognitively normal adults earlier in the aging process. Individuals who experienced childhood disadvantage manifested greater vulnerability to cortical microstructure-related visual memory dysfunction than their non-disadvantaged counterparts who exhibited resilience in the face of low cortical microstructural integrity.


The functions of type I and type II natural killer T cells in inflammatory bowel diseases.
CD1d-restricted natural killer T (NKT) cells are a distinct subset of T cells that rapidly produce an array of cytokines on activation and play a critical role in regulating various immune responses. NKT cells are classified into 2 groups based on differences in T-cell receptor usage. Type I NKT cells have an invariant T-cell receptor α-chain and are readily detectable by α-galactosylceramide (α-GalCer)-loaded CD1d tetramers. Type II NKT cells have a more diverse T-cell receptor repertoire and cannot be directly identified. Both types of NKT cells and multiple CD1d-expressing cell types are present in the intestine, and their interactions are likely to be modulated by pathogenic and commensal microbes, which in turn contribute to the intestinal immune responses in health and disease. Indeed, in several animal models of inflammatory bowel disease, type I NKT cells have been shown to make both protective and pathogenic contributions to disease. In contrast, in patients with ulcerative colitis, and a mouse model in which both CD1d expression and the frequency of type II NKT cells are increased, type II NKT cells seem to promote intestinal inflammation. In this review, we summarize the present knowledge on the antigen recognition, activation, and function of NKT cells with a particular focus on their role in inflammatory bowel disease and discuss factors that may influence the functional outcome of NKT cell responses in intestinal inflammation.


End-Point Prediction of Granule Moisture in a ConsiGma™-25 Segmented Fluid Bed Dryer.
Continuously operated pharmaceutical manufacturing lines often consist of a wet granulation unit operation, followed by a (semi-) continuous dryer. The operating conditions of the dryer are crucial for obtaining a desired final granule moisture. Commercially available dryers lack of a thorough online measurement of granule moisture during the drying process. However, this information could improve the operation of the equipment considerably, yielding a granule moisture close to the desired value (e.g., by drying time and process parameter adjustments in real-time). The paper at hand proposes a process model, which can be parameterized from a very limited number of experiments and then be used as a so-called soft sensor for predicting granule moisture. It utilizes available process measurements for the estimation of the granule moisture. The development of the model as well as parameter identification and validation experiments are provided. The proposed model paves the way for the application of sophisticated observer concepts. Possible future activities on that topic are outlined in the paper.


PubChem: a public information system for analyzing bioactivities of small molecules.
PubChem (http://pubchem.ncbi.nlm.nih.gov) is a public repository for biological properties of small molecules hosted by the US National Institutes of Health (NIH). PubChem BioAssay database currently contains biological test results for more than 700 000 compounds. The goal of PubChem is to make this information easily accessible to biomedical researchers. In this work, we present a set of web servers to facilitate and optimize the utility of biological activity information within PubChem. These web-based services provide tools for rapid data retrieval, integration and comparison of biological screening results, exploratory structure-activity analysis, and target selectivity examination. This article reviews these bioactivity analysis tools and discusses their uses. Most of the tools described in this work can be directly accessed at http://pubchem.ncbi.nlm.nih.gov/assay/. URLs for accessing other tools described in this work are specified individually.


Central nervous system toxicity and cerebrospinal fluid pharmacokinetics of intraventricularly administered bleomycin in beagles.
The neurotoxic effects and cerebrospinal fluid (CSF) pharmacokinetics of bleomycin were evaluated in beagles after chronic intraventricular administration twice a week for 8 consecutive weeks. Bleomycin was reasonably well tolerated at doses of 0.067 to 0.3 mg/week. Doses higher than 0.3 mg/week produced marked elevation of CSF protein levels and a necrotizing vasculitis within the central nervous system. Pharmacokinetic studies were performed approximately 1 month after the completion of the toxicity studies. [3H]inulin was used as a reference compound. Both [3H]inulin and bleomycin were cleared from the CSF more slowly than in previous studies and more slowly than in normal dogs, which suggests that bulk CSF absorption was reduced by the drug, probably secondary to protein-induced blockage of the arachnoid granulations through which CSF is normally absorbed. Because a "minimally toxic" dose of bleomycin (approximately 0.1 mg/week) produces a CSF C X t of only 1.9 mg/min/ml, we believe that a Phase I clinical trial would be too dangerous given the limited therapeutic potential that a dose of 0.1 mg/week could achieve.


Donepezil effects on hippocampal and prefrontal functional connectivity in Alzheimer's disease: preliminary report.
We used functional connectivity magnetic resonance imaging (fcMRI) to investigate changes in interhemispheric brain connectivity in 11 patients with mild Alzheimer's disease (AD) following eight weeks of treatment with the cholinesterase inhibitor donepezil. We examined functional connectivity between four homologous temporal, frontal, and occipital regions. These regions were selected to represent sites of AD neuropathology, sites of donepezil-related brain activation change in prior studies, and sites that are minimally affected by the pathologic changes of AD. Based on previous findings of selective, localized frontal responses to donepezil, we predicted that frontal connectivity would be most strongly impacted by treatment. Of the areas examined, we found that treatment had a significant effect only on functional connectivity between right and left dorsolateral prefrontal cortices. Implications for understanding the impact of donepezil treatment on brain functioning and behavior in patients with AD are discussed. This preliminary report suggests that fcMRI may provide a useful index of treatment outcome in diseases affecting brain connectivity. Future research should investigate these treatment-related changes in larger samples of patients and age-matched controls.


[Expert evaluation in bony injuries of the spine].
The conventional way of appreciating working capacity is no longer satisfying considering the improved diagnostic tools with increasing frequency of various deformities, instability and ankylosis and their combinations. As in the evaluation of peripheral joint function a concept is proposed, which is based on the structural changes in the motion segments. The concept is practicable in all parts of the vertebral column and allows a differentiated judgement within the usual percentages.


A comparative study of nebivolol and (S) atenolol on blood pressure and heart rate on essential hypertensive patients.
To study the effect of nebivolol 5 mg once daily versus (S)-atenolol 25 mg once daily in patients with essential hypertension. A prospective study was conducted at RLJH and Research Centre which included 30 patients in each group with essential hypertension. The sex, age, presenting illness, and family history of the patients were recorded. Investigations such as blood sugar, urine analysis, kidney function test, lipid profile, and ECG were performed before starting the treatment. Any adverse effects during the treatment were noted. Blood pressure and heart rate were recorded at baseline and during follow-up. One group received nebivolol 5 mg once daily and other group (S)atenolol 25 mg once daily. Patients were followed-up every 15 days for 3 months. Nebivolol group had 18 males and 12 females with mean age 50.6 ± 9.5 years, (S)-atenolol had 16 males and 14 females with mean age 54.4 ± 9 years. Patients receiving nebivolol and (S)-atenolol showed a significant fall (P <·0001) in systolic (SBP), diastolic blood pressure (DBP), and heart rate at the end of first, second, and third month when compared to baseline. The difference in fall in SBP and DBP was insignificant between the groups, but fall in heart rate was significant (P <·0001). Adverse effects such as headache, dizziness, and fatigue were reported with both drugs. Reduction of blood pressure with nebivolol and (S)atenolol was similar, but fall in blood pressure from baseline was highly significant in both groups.


Clinical characteristics, treatment and prognosis of angiocentric glioma.
Angiocentric glioma (AG) is a rare subtype of neuroepithelial tumor in children and young adults that commonly presents with seizures. To study the clinical characteristics, treatment and prognosis of patients with AG, the features of two cases of AG were described and 108 cases reported in the literature were assessed. The cases of the present study were two males aged 8 and 16 years, who mainly presented with seizures. MRI revealed superficial, non-enhanced lesions in the left temporal and right frontal lobe, respectively. The two patients underwent gross total resection (GTR) and remained seizure-free without neurological deficits after 3.5 and 2.5 years, respectively. Histopathological examination revealed that the tumors consisted of monomorphous cells that surrounded the blood vessels and neurons in the cerebral cortex, and formed concentric sleeves or pseudorosettes. Furthermore, immunostaining indicated that the diffuse infiltrative neoplastic cells were positive for glial fibrillary acidic protein and a dot-like pattern of epithelial membrane antigen was observed. AG mostly appeared similar to low-grade gliomas on MRI. GTR of the lesions was curative and radiation or chemotherapy were not required. AG typically has a favorable prognosis, with low mortality and incidence of disability.


Study of polyacrylamide grafted starch based algal flocculation towards applications in algal biomass harvesting.
Microalgae may be the source of high amount of lipid and protein. It has the property for carbon dioxide sequestration, recycling and also can remove pollutants from wastewater. Using traditional methods, collection of algal biomass is either cost effective, time consuming or may be toxic due to use of chemical salts. The aim of this study is to harvest freshwater microalgae (Chlorella sp. CB4) biomass by using polymer. Polyacrylamide grafted starch (St-g-PAM) has been synthesized by microwave assisted method involving a synergism of microwave radiation and ceric ammonium nitrate (CAN) to initiate the grafting reaction. The synthesis was optimized in terms of CAN and monomer (acrylamide) concentration. The algal flocculation efficacy of all the grades of this graft copolymer was studied through standard 'Jar test' procedure. Effects of percentage grafting, pH and zeta potential on percentage recovery of algal biomass were thoroughly investigated.


Recent progress and challenges in nanotechnology for biomedical applications: an insight into the analysis of neurotransmitters.
Nanotechnology offers exciting opportunities and unprecedented compatibilities in manipulating chemical and biological materials at the atomic or molecular scale for the development of novel functional materials with enhanced capabilities. It plays a central role in the recent technological advances in biomedical technology, especially in the areas of disease diagnosis, drug design and drug delivery. In this review, we present the recent trend and challenges in the development of nanomaterials for biomedical applications with a special emphasis on the analysis of neurotransmitters. Neurotransmitters are the chemical messengers which transform information and signals all over the body. They play prime role in functioning of the central nervous system (CNS) and governs most of the metabolic functions including movement, pleasure, pain, mood, emotion, thinking, digestion, sleep, addiction, fear, anxiety and depression. Thus, development of high-performance and user-friendly analytical methods for ultra-sensitive detection of neurotransmitters remain a major challenge in modern biomedical analysis. Nanostructured materials are emerging as a powerful mean for diagnosis of CNS disorders because of their unique optical, size and surface characteristics. This review provides a brief outline on the basic concepts and recent advancements of nanotechnology for biomedical applications, especially in the analysis of neurotransmitters. A brief introduction to the nanomaterials, bionanotechnology and neurotransmitters is also included along with discussions on most of the patents published in these areas.


[Perioperative monitoring of the course of anesthesia, the postanesthesia visit and inquiry of patient satisfaction. A prospective study of parameters in process and outcome quality in anesthesia].
This study is an investigation into the results of reporting on incidents during and after anaesthesia, to reveal any possible associations between intra-procedural and final outcome. The study contributes to the quality assurance project of the German Society of Anaesthesiology and Intensive Care (DGAI). We adjusted and tabulated our data for preoperative risk and for different methods of anaesthesia. This nation-wide DGAI project was launched to compare clinical institutions in accordance with German social legislation. DGAI recommends standardised documentation of parameters representing quality of process (QP) defined by a 63-items list, during every anaesthesia course. Additionally, quality of outcome parameters (QO) defined by a 64-items list should be assessed by an anaesthetist during a standardised postoperative ward round by means of spot checks. A questionnaire covering subjective complaints (SC) and patient satisfaction, is optional. The combination of these tools was evaluated during a 5-month period in every patient (n = 282) on a traumatological surgery ward. QP and QO showed a significant association (p < 0.0001). The incidence of both parameters increases according to risk factors like age and ASA physical status. This was not significant in respect of the type of anaesthesia (p = 0.20). Whereas perioperative QP parameters were less frequent in regional versus general anaesthesia (p < 0.0001), there was no postoperative difference in QO as seen by the anaesthetist (p = 0.20). However, postoperative SC were less frequent with regional anaesthesia (p < 0.0001). The association of QO and SC was mot significant (p = 0.24). There was comparable preoperative morbidity (p = 0.74) for both anaesthetic procedures. 96.5% of all patients expressed overall satisfaction. Despite this fact, nausea (25%), vomiting (29%), thurst (29%) and particularly wound pain (33%) were frequent. Quality parameters assessed by anaesthetists and patients are independent in respect to their frequency. For this purpose, anaesthesiological quality assurance must focus on both the anaesthetist and the patient. As clinical consequence, we suggest establishing an interdisciplinary post-anaesthesia service. Acceptance by, and collaboration between, the surgical disciplines are indispensable especially for a successful application of effective pain and antiemetic therapy.


Ten Years Follow Up of Patients with Tinnitus and Normal Hearing.
To describe the natural history of tinnitus in normal hearing patients. This is a prospective longitudinal observational study. Tertiary University Hospital. In 2009 we studied 68 patients with significant tinnitus and normal hearing (Study Group-SG) compared to 46 patients with normal hearing and no tinnitus (Control Group-CG). We have been able to contact and recruit 24 patients from SG (35.3 %) and ten patients from CG) (21.74%) in 2019. Both groups were submitted to conventional audiometry, Distortion Product Otoacoustic Emissions (DPOAE), Tinnitus Handicap Inventory (THI), Visual Analog Scale (VAS) and Beck`S Depression and Anxiety Scales (BDS, BAS). These measures were compared with the same ones performed ten years ago in the same groups of patients. The software SPSS for windows version 21 was used. P values <0.05 were considered significant. Student t test was used to compare SG and CG results in 2019 and to compare 2009 and 2019 outcomes.. Hearing loss, especially in the higher frequencies, were significantly worse in SG compared to GC in 2019. DPOAE were significantly worse in SG and CG in 2019 however CG had more abnormal results than SG. These results suggest that outer hair cells lesions are not the only determinant for tinnitus and hearing loss development. These results suggest that outer hair cells lesions are not the only determinant for tinnitus and hearing loss development.


Attitudes and Intentions of US Veterans Regarding COVID-19 Vaccination.
Compared with the general population, veterans are at high risk for COVID-19 and have a complex relationship with the government. This potentially affects their attitudes toward receiving COVID-19 vaccines. To assess veterans' attitudes toward and intentions to receive COVID-19 vaccines. This cross-sectional web-based survey study used data from the Department of Veterans Affairs (VA) Survey of Healthcare Experiences of Patients' Veterans Insight Panel, fielded between March 12 and 28, 2021. Of 3420 veterans who were sent a link to complete a 58-item web-based survey, 1178 veterans (34%) completed the survey. Data were analyzed from April 1 to August 25, 2021. Veterans eligible for COVID-19 vaccines. The outcomes of interest were veterans' experiences with COVID-19, vaccination status and intention groups, reasons for receiving or not receiving a vaccine, self-reported health status, and trusted and preferred sources of information about COVID-19 vaccines. Reasons for not getting vaccinated were classified into categories of vaccine deliberation, dissent, distrust, indifference, skepticism, and policy and processes. Among 1178 respondents, 974 (83%) were men, 130 (11%) were women, and 141 (12%) were transgender or nonbinary; 58 respondents (5%) were Black, 54 veterans (5%) were Hispanic or Latino, and 987 veterans (84%) were non-Hispanic White. The mean (SD) age of respondents was 66.7 (10.1) years. A total of 817 respondents (71%) self-reported being vaccinated against COVID-19. Of 339 respondents (29%) who were not vaccinated, those unsure of getting vaccinated were more likely to report fair or poor overall health (32 respondents [43%]) and mental health (33 respondents [44%]) than other nonvaccinated groups (overall health: range, 20%-32%; mental health: range, 18%-40%). Top reasons for not being vaccinated were skepticism (120 respondents [36%] were concerned about side effects; 65 respondents [20%] preferred using few medications; 63 respondents [19%] preferred gaining natural immunity), deliberation (74 respondents [22%] preferred to wait because vaccine is new), and distrust (61 respondents [18%] did not trust the health care system). Among respondents who were vaccinated, preventing oneself from getting sick (462 respondents [57%]) and contributing to the end of the COVID-19 pandemic (453 respondents [56%]) were top reasons for getting vaccinated. All veterans reported the VA as 1 of their top trusted sources of information. The proportion of respondents trusting their VA health care practitioner as a source of vaccine information was higher among those unsure about vaccination compared with those who indicated they would definitely not or probably not get vaccinated (18 respondents [26%] vs 15 respondents [15%]). There were no significant associations between vaccine intention groups and age (χ24 = 5.90; P = .21) or gender (χ22 = 3.99; P = .14). These findings provide information needed to develop trusted messages used in conversations between VA health care practitioners and veterans addressing specific vaccine hesitancy reasons, as well as those in worse health. Conversations need to emphasize societal reasons for getting vaccinated and benefits to one's own health.


Immunodetection of human telomerase reverse-transcriptase (hTERT) re-appraised: nucleolin and telomerase cross paths.
The involvement of telomerase in cellular immortalization and senescence has often been assessed by means of telomerase expression at the RNA level and quantification of telomerase activity by the telomeric repeat amplification protocol assay. However, these methods either neglected the existence of various telomerase splice variants, or ignored the nonconventional functions of telomerase independent of its ability to elongate and maintain telomere length. Immunodetection of telomerase is now being recognized as a necessary approach to precisely elucidate its roles in oncogenesis and senescence. A few antibodies directed against the catalytic subunit of the human telomerase (hTERT) are currently used but their specificity is not always demonstrated. A survey of the literature showed inconsistencies and led us to comparatively re-evaluate the most frequently used antibodies. Surprisingly, mass spectrometry, two-dimensional gel analysis and immunofluorescent experiments revealed that the most frequently used hTERT immunoprobe, a mouse monoclonal antibody that was claimed to be directed against an hTERT protein epitope, in fact recognizes nucleolin rather than telomerase. Our findings have interesting implications regarding the biology of nucleolin and telomerase in the context of pathophysiological investigations recently carried out.


Bone marrow: the bursa equivalent in man?
Human bone marrow lymphoid cells, particularly when enriched with plasma cells, as in multiple myeloma, respond to pokeweed mitogen and to antiserum to immunoglobulin but not to phytohemagglutinin. Cells of patients with X-linked agammaglobulinemia of the bursal deficient type failed to respond to either pokeweed or to the antiserum to immunoglobulin. Leukocytes of the agammaglobulinemia patients however responded in a normal fashion to phytohemagglutinin. Just as the in vitro response to phytohemagglutinin is taken as an index of the thymus-dependent system, the in vitro response to both antiserum to immunoglobulin and pokeweed may be considered an index for the bursaldependent system. Human bone marrow, therefore, contains bursal cells and probably very few or no thymus cells.


A Wearable Cellulose Acetate-Coated Mouthguard Biosensor for In Vivo Salivary Glucose Measurement.
In this study, a cellulose acetate (CA) membrane is formed as an interference rejection membrane on a glucose sensor to measure glucose in saliva. Glucose in saliva is successfully measured in vivo without any pretreatment of human saliva. A mouthguard (MG) glucose sensor is developed to monitor salivary glucose, which is reported to be correlated with the blood glucose level. Salivary components of ascorbic acid (AA) and uric acid (UA) hinder the accurate measurement of the glucose concentration of human saliva. CA-coated electrodes are prepared to investigate the interference rejection membrane. To measure hydrogen peroxide, which is a reaction product of glucose oxidase, effects of AA and UA are examined. Characteristics of the fabricated biosensor are examined on the basis of artificial saliva. The as-developed MG sensor can quantify the glucose concentration in the range of 1.75-10 000 μmol/L, which includes a salivary sugar concentration of 20-200 μmol/L. For the measurement of saliva samples collected from healthy subjects, the output corresponding to the concentration is confirmed; this suggests the possibility of glucose measurement. This MG glucose sensor can provide a useful method for the unrestricted and noninvasive monitoring of saliva glucose for the management of diabetes patients.


The Japanese cord blood bank network experience with cord blood transplantation from unrelated donors for haematological malignancies: an evaluation of graft-versus-host disease prophylaxis.
Cryopreserved umbilical cord blood (CB) from unrelated donors can restore haematopoiesis after myeloablative therapy in patients with haematological malignancy. We investigated the clinical outcomes of CB transplantation (CBT) with special emphasis on graft-versus-host disease (GVHD) prophylaxis. Patients with haematological malignancies (n = 216) received intensive chemotherapy or immunosuppressive therapy, followed by transplantation of cryopreserved CB cells from unrelated donors. The clinical outcomes, i.e. haematological reconstitution, the incidence of acute or chronic GVHD, relapse and event-free survival (EFS), were evaluated. The estimated probability of neutrophil recovery was 88.2%. The median follow-up for the survivors was 557 d (range 21-1492 d). The overall and EFS rates were 32.6% and 25.5%, respectively, 3.5 years after transplantation. Multivariate analysis using Cox's proportional hazards model showed that high-risk disease status at CBT and single-drug GVHD prophylaxis were associated with worse 2-year EFS rates [P = 0.0013, relative risk (RR) 1.90, 95% confidence interval (CI) 1.28-2.81 and P = 0.0007, RR 1.91, 95% CI 1.31-2.79 respectively). Age at CBT had no significant influence on EFS. Cryopreserved CB from unrelated donors can restore haematopoiesis in patients with haematological malignancy. Although the incidence is low, the prophylaxis for acute GVHD is an important factor for survival of CBT from unrelated donors. A high rate of suitable donors was found, with a probability of 1 to every 18 CB units, when compared with human leucocyte antigen matching at other haematopoietic stem cell banks.


Effects of yoga on depressive symptoms in people with mental disorders: a systematic review and meta-analysis.
To assess whether physically active yoga is superior to waitlist control, treatment as usual and attention control in alleviating depressive symptoms in people with a diagnosed mental disorder recognised by the Diagnostic and Statistical Manual of Mental Disorders (DSM). Systematic review and meta-analysis following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Data were obtained from online databases (MEDLINE, EMBASE, PsychINFO, CENTRAL, EMCARE, PEDro). The search and collection of eligible studies was conducted up to 14 May 2019 (PROSPERO registration No CRD42018090441). We included randomised controlled trials with a yoga intervention comprising ≥50% physical activity in adults with a recognised diagnosed mental disorder according to DSM-3, 4 or 5. 19 studies were included in the review (1080 participants) and 13 studies were included in the meta-analysis (632 participants). Disorders of depression, post-traumatic stress, schizophrenia, anxiety, alcohol dependence and bipolar were included. Yoga showed greater reductions in depressive symptoms than waitlist, treatment as usual and attention control (standardised mean difference=0.41; 95% CI -0.65 to -0.17; p<0.001). Greater reductions in depressive symptoms were associated with higher frequency of yoga sessions per week (β=-0.44, p<0.01).


[Goiter and pregnancy].
The author refers to the changes occurred in thyroid dimensions during pregnancy, evaluated clinically and echographically. He also describes the changes in thyroid normal function on pregnant women and the different stages of the fetal thyroid maturation. Some complementary tests must be carried out, when there is a presence of a goitre nodule, which opportunity and relevance the need are discussed. The possibility of thyroid carcinoma is referred as its most adequate treatment.


Development of MijnAVL, an Interactive Portal to Empower Breast and Lung Cancer Survivors: An Iterative, Multi-Stakeholder Approach.
MijnAVL (MyAVL) is an interactive portal being developed to empower cancer survivors. Literature review and focus groups yielded the selection of features such as access to the electronic medical record (EMR), patient reported outcomes (PROs) and related feedback, and a physical activity support program. Our aim was to present a final design of MijnAVL based on (1) health professionals' evaluation of proposed features, (2) cancer survivors' evaluation of a first draft, and (3) cancer survivors' evaluation of a functional online prototype. Professionals from various disciplines gave input to the content of and procedures related to MijnAVL. Subsequently, 16 cancer survivors participated in an interview to evaluate content and graphic design of a first draft (shown with screenshots). Finally, 7 survivors participated in a usability test with a fully functional prototype. They performed predefined tasks (eg, logging in, finding a test result, completing a questionnaire) while thinking aloud. Descriptive statistics and simple content analysis were used to analyze the data of both the interviews and the usability tests. Professionals supported access to the EMR (eg, histology reports, lab results, and their letters to general practitioners). They also informed the development of PROs and the physical activity support program. Based on the first draft, survivors selected the preferred graphic design, approved the features and provided suggestions for the content (eg, explanation of medical jargon, more concise texts, notification by emails). Usability tests revealed that it was relatively easy to navigate the website and use the different features. Recommendations included, among others, a frequently asked questions section and the use of hyperlinks between different parts of the website. The development of MijnAVL, an interactive portal to empower breast and lung cancer survivors, was performed iteratively and involved multiple groups of end-users. This approach resulted in a usable and understandable final version. Its effectiveness should be determined in further research.


Role of glucocorticoids in activation of hepatic PEPCK gene transcription during exercise.
The objective of these studies was to determine the molecular basis for the activation of phosphoenolpyruvate carboxykinase (PEPCK) gene transcription during prolonged submaximal exercise. Mice were fed a high-carbohydrate diet for 1 wk and exercised continuously by swimming for up to 120 min. The level of hepatic PEPCK mRNA increased progressively during exercise, reaching 510% above control, whereas transcription of the PEPCK gene increased 1,000%, before decreasing to control levels within 60 min of recovery. In transgenic mice carrying a chimeric gene consisting of the PEPCK promoter linked to a reporter gene for bovine growth hormone (bGH), PEPCK(-460)-bGH, the level of hepatic bGH mRNA increased by 490% in response to exercise, similar to the increase in the expression of the native PEPCK gene. However, in transgenic mice with a deletion of the glucocorticoid regulatory unit, PEPCK(-355)-bGH, bGH mRNA did not increase above control values. In transgenic mice with a block mutation in adenosine 3',5'-cyclic monophosphate (cAMP) regulatory regions -90/-82 and -250/-234, PEPCK cAMP response element 1 (CRE-1)/P3(1)-bGH, exercise increased bGH mRNA 260% above controls. Adrenalectomy (Adx) had no effect on PEPCK mRNA levels in nonexercised mice, whereas in adrenalectomized (Adx)-exercised mice, PEPCK mRNA increased only 80% above basal, and, in Adx mice injected with dexamethasone, PEPCK mRNA increased with exercise 570% above controls. Exercise was also associated with a large increase in transcription of the gene for the transcription factor CCAAT/enhancer-binding protein beta (C/EBP-beta) and a smaller rise in transcription of c-jun gene, both of which returned to control levels during recovery.(ABSTRACT TRUNCATED AT 250 WORDS)


Binding of MBD proteins to DNA blocks Tet1 function thereby modulating transcriptional noise.
Aberrant DNA methylation is a hallmark of various human disorders, indicating that the spatial and temporal regulation of methylation readers and modifiers is imperative for development and differentiation. In particular, the cross-regulation between 5-methylcytosine binders (MBD) and modifiers (Tet) has not been investigated. Here, we show that binding of Mecp2 and Mbd2 to DNA protects 5-methylcytosine from Tet1-mediated oxidation. The mechanism is not based on competition for 5-methylcytosine binding but on Mecp2 and Mbd2 directly restricting Tet1 access to DNA. We demonstrate that the efficiency of this process depends on the number of bound MBDs per DNA molecule. Accordingly, we find 5-hydroxymethylcytosine enriched at heterochromatin of Mecp2-deficient neurons of a mouse model for Rett syndrome and Tet1-induced reexpression of silenced major satellite repeats. These data unveil fundamental regulatory mechanisms of Tet enzymes and their potential pathophysiological role in Rett syndrome. Importantly, it suggests that Mecp2 and Mbd2 have an essential physiological role as guardians of the epigenome.


[Not everything is Zika: congenital toxoplasmosis, still prevalent in Colombia?]
Congenital toxoplasmosis continues to be a public health threat. Even existing guidelines, publicly known, its implementation and lack of appropriate interpretation of serological tests in pregnancy is often observed. This leds to failure in opportunities for positive and known interventions to decrease the fetal risk due to Toxoplasma gondii infection. We reported herein a case series, with variable neurological and systemic compromise (respiratory distress, hepatosplenomegaly, enterocolitis, brain calcifications, thrombocytopenia, ascites, shock), even fatal, calling for awareness about the fact that despite the Zika epidemics in 2015-2016 in Brazil, Colombia and other countries, precisely toxoplasmosis, is a differential diagnosis still prevalent in these territories, that can leds to severe consequences, with neurological disability and risk of ocular damage, even lately. Additionally, with varieties of T. gondii with more aggressive patterns in Latin America, which make worse those cases, including also a higher risk of death.


Task-related BOLD responses and resting-state functional connectivity during physiological clamping of end-tidal CO(2).
Carbon dioxide (CO(2)), a potent vasodilator, is known to have a significant impact on the blood-oxygen level dependent (BOLD) signal. With the growing interest in studying synchronized BOLD fluctuations during the resting state, the extent to which the apparent synchrony is due to variations in the end-tidal pressure of CO(2) (PETCO(2)) is an important consideration. CO(2)-related fluctuations in BOLD signal may also represent a potential confound when studying task-related responses, especially if breathing depth and rate are affected by the task. While previous studies of the above issues have explored retrospective correction of BOLD fluctuations related to arterial PCO(2), here we demonstrate an alternative approach based on physiological clamping of the arterial CO(2) level to a near-constant value. We present data comparing resting-state functional connectivity within the default-mode-network (DMN), as well as task-related BOLD responses, acquired in two conditions in each subject: 1) while subject's PETCO(2) was allowed to vary spontaneously; and 2) while controlling subject's PETCO(2) within a narrow range. Strong task-related responses and areas of maximal signal correlation in the DMN were not significantly altered by suppressing fluctuations in PETCO(2). Controlling PETCO(2) did, however, improve the performance of retrospective physiological noise correction techniques, allowing detection of additional regions of task-related response and resting-state connectivity in highly vascularized regions such as occipital cortex. While these results serve to further rule out systemic physiological fluctuations as a significant source of apparent resting-state network connectivity, they also demonstrate that fluctuations in arterial CO(2) are one of the factors limiting sensitivity in task-based and resting-state fMRI, particularly in regions of high vascular density. This must be considered when comparing subject groups who might exhibit differences in respiratory physiology or breathing patterns.


A randomized study of thromboprophylaxis in women with unexplained consecutive recurrent miscarriages.
To compare the effect of aspirin and enoxaparin on live births in women with unexplained recurrent miscarriages, as well as secondary outcomes including birth weight, uterine and umbilical blood flows, and congenital malformations. Multicenter randomized comparative cohort study. Four centers including two university hospitals, a peripheral general hospital, and a community health clinic. One hundred seven patients were randomized, 104 were available for analysis; 54 were randomized to enoxaparin and 50 to aspirin. Treatment with enoxaparin or aspirin in subsequent pregnancy. Subsequent live births or miscarriage, and the incidence of obstetric complications. Both groups had a similar live birth rate (relative risk = 0.92, 95% confidence interval: 0.58-1.46). In primary aborters, live births occurred in 17 of 18 (94%) enoxaparin-treated pregnancies compared to 18 of 22 (81%) aspirin-treated pregnancies. In the aspirin group, two pregnancies were terminated: for tricuspid insufficiency and for hemolysis, elevated liver enzymes, low platelet (HELLP) syndrome. One enoxaparin-treated infant was growth restricted (2,020 g) at 36 weeks. Preeclampsia was found in three aspirin-treated patients. Preterm delivery, placental Doppler blood flow, apgar scores, and mean birth weights were similar in both groups. In the aspirin group, one infant underwent orchidectomy after testicular torsion in utero, and one infant had hypoglycemia and convulsions. Both regimens were associated with a high live birth rate and few late pregnancy complications.


Bordetella pertussis adenylate cyclase toxin modulates innate and adaptive immune responses: distinct roles for acylation and enzymatic activity in immunomodulation and cell death.
Adenylate cyclase toxin (CyaA) of Bordetella pertussis belongs to the repeat in toxin family of pore-forming toxins, which require posttranslational acylation to lyse eukaryotic cells. CyaA modulates dendritic cell (DC) and macrophage function upon stimulation with LPS. In this study, we examined the roles of acylation and enzymatic activity in the immunomodulatory and lytic effects of CyaA. The adenylate cyclase activity of CyaA was necessary for its modulatory effects on murine innate immune cells. In contrast, acylation was not essential for the immunomodulatory function of CyaA, but was required for maximal caspase-3 activation and cytotoxic activity. The wild-type acylated toxin (A-CyaA) and nonacylated CyaA (NA-CyaA), but not CyaA with an inactive adenylate cyclase domain (iAC-CyaA), enhanced TLR-ligand-induced IL-10 and inhibited IL-12, TNF-alpha, and CCL3 production by macrophages and DC. In addition, both A-CyaA and NA-CyaA, but not iAC-CyaA, enhanced surface expression of CD80 and decreased CpG-stimulated CD40 and ICAM-1 expression on immature DC. Furthermore, both A-CyaA and NA-CyaA promoted the induction of murine IgG1 Abs, Th2, and regulatory T cells against coadministered Ags in vivo, whereas iAC-CyaA had more limited adjuvant activity. In contrast, A-CyaA and iAC-CyaA induced caspase-3 activation and cell death in macrophages, but these effects were considerably reduced or absent with NA-CyaA. Our findings demonstrate that the enzymatic activity plays a critical role in the immunomodulatory effects of CyaA, whereas acylation facilitates the induction of apoptosis and cell lysis, and as such, NA-CyaA has considerable potential as a nontoxic therapeutic molecule with potent anti-inflammatory properties.


Expression of the HSP 70.1 gene, a landmark of early zygotic activity in the mouse embryo, is restricted to the first burst of transcription.
Activation of the mouse embryonic genome at the 2-cell stage is characterized by the synthesis of several alpha-amanitin-sensitive polypeptides, some of which belong to the multigenic hsp 70 family. In the present work we show that a member of this family, the HSP 70.1 gene, is highly transcribed at the onset of zygotic genome activation. Transcription of this gene began as early as the 1-cell stage. Expression of the gene continued through the early 2-cell stage but was repressed before the completion of the second round of DNA replication. During this period we observed that the level of transcription was modulated by in vitro culture conditions. The coincidence of repression of HSP70.1 transcription with the second round of DNA replication was not found for other transcription-dependent polypeptides synthesized at the 2-cell stage.


Maternal erythrocyte ENT1-mediated AMPK activation counteracts placental hypoxia and supports fetal growth.
Insufficient O2 supply is frequently associated with fetal growth restriction (FGR), a leading cause of perinatal mortality and morbidity. Although the erythrocyte is the most abundant and only cell type to deliver O2 in our body, its function and regulatory mechanism in FGR remain unknown. Here, we report that genetic ablation of mouse erythrocyte equilibrative nucleoside transporter 1 (eENT1) in dams, but not placentas or fetuses, results in FGR. Unbiased high-throughput metabolic profiling coupled with in vitro and in vivo flux analyses with isotopically labeled tracers led us to discover that maternal eENT1-dependent adenosine uptake is critical in activating AMPK by controlling the AMP/ATP ratio and its downstream target, bisphosphoglycerate mutase (BPGM); in turn, BPGM mediates 2,3-BPG production, which enhances O2 delivery to maintain placental oxygenation. Mechanistically and functionally, we revealed that genetic ablation of maternal eENT1 increases placental HIF-1α; preferentially reduces placental large neutral aa transporter 1 (LAT1) expression, activity, and aa supply; and induces FGR. Translationally, we revealed that elevated HIF-1α directly reduces LAT1 gene expression in cultured human trophoblasts. We demonstrate the importance and molecular insight of maternal eENT1 in fetal growth and open up potentially new diagnostic and therapeutic possibilities for FGR.


Interlaminar shear stresses and laminae separation in a disc. Finite element analysis of the L3-L4 motion segment subjected to axial compressive loads.
This study analyzed interlaminar shear stresses across the laminae of a ligamentous L3-L4 motion segment. A three-dimensional finite element model of the motion segment was developed and its response in axial compression mode was predicted. The contributions of "mechanical" factors in producing laminae separation in a disc are not well understood, especially when the nucleus is still gel-like in appearance (stage 1 of disc degeneration). All types of stresses are likely to contribute to laminae separation. The authors believe it is partially due to the interlaminar shear stresses at the laminae interfaces in specific regions of an intact disc because the disc is a composite structure. The effects of anular tears on the interlaminar shear stresses were also investigated. These tears can be circumferential or radial in nature, and commonly occur in the aged, degenerated disc. A large number of biomechanical studies dealing with the role of the disc vis-a-vis other spinal components have been reported in the literature. The role of mechanical factors, however, in producing laminae separation, especially when the nucleus is still gel-like in appearance (stage 1 of disc degeneration), is not entirely clear. A three-dimensional nonlinear finite element model of an intact L3-L4 lumbar motion segment, based on the use of a special type of element for the disc anulus, was created to investigate the interlaminar shear stresses in the anulus. The effects of radial "out-in," radial "in-out," and "circumferential" injuries were analyzed. Injury was modeled as element removal in the posterolateral portion of the disc. Models subjected to axial compressive loads, ranging from 200 N to 2000 N, were analyzed. In addition to the interlaminar shear stresses, disc bulge, and displacements including coupled motions were predicted. The theoretical disc bulge predictions for the radial in-out injury case were in agreement with the disc bulge data obtained experimentally. Displacements, disc bulge, and coupled motions increased with injury, as expected. The interlaminar shear stresses were highest in the posterolateral portions of the intact disc model. Interlaminar shear stresses, in general, increased with injury. Also, a slight increase in circumferential injury was sufficient to see a substantial increase in interlaminar shear stresses. The interlaminar shear stresses being higher in the posterolateral regions of the intact disc reinforces that, from clinical studies, tears originate in the posterolateral portion of the disc. The large interlaminar shear stresses, caused by asymmetry in the disc structure due to injury, along with chemical and structural changes in the disc with age, may be an important cause of further degeneration through laminae separation. This is the case for traditional composite laminates. This study points out the importance of interlaminar shear stresses to gain further understanding of the role of mechanical factors in producing disc degeneration, especially delamination of the anulus. Clinical relevance of the findings and possible relationship to the aging process are explored.


Supporting Future Cannabis Policy - Developing a Standard Joint Unit: A Brief Back-Casting Exercise.
The standardization of cannabis doses is a priority for research, policy-making, clinical and harm-reduction interventions and consumer security. Scientists have called for standard units of dosing for cannabis, similar to those used for alcohol. A Standard Joint Unit (SJU) would facilitate preventive and intervention models in ways similar to the Standard Drink (SD). Learning from the SD experiences allows researchers to tackle emerging barriers to the SJU by applying modern forecasting methods. During a workshop at the Lisbon Addictions Conference 2019, a back-casting foresight method was used to address challenges and achieve consensus in developing an SJU. Thirty-two professionals from 13 countries and 10 disciplines participated. Descriptive analysis of the workshop was carried out by the organizers and shared with the participants in order to suggest amendments. Several characteristics of the SJU were defined: (1) core values: easy-to use, universal, focused on THC, accurate, and accessible; (2) key challenges: sudden changes in patterns of use, heterogeneity of cannabis compounds as well as in administration routes, variations over time in THC concentrations, and of laws that regulate the legal status of recreational and medical cannabis use); and (3) facilitators: previous experience with standardized measurements, funding opportunities, multi-stakeholder support, high prevalence of cannabis users, and widespread changes in legislation. Participants also identified three initial steps for the implementation of a SJU by 2030: (1) Building a task-force to develop a consensus-based SJU; (2) Expanded available national-level data; (3) Linking SJU consumption to the concept of "risky use," based on evidence of harms.


[Role of supravesicular urine diversion in the treatment of patients with renal and urethral tuberculosis].
The article presents the results of the survey, etiopathogenetic and surgery treatment of 73 patients with nephrotuberculosis complicated by tuberculous ureteritis. Patients were divided into 4 groups. 1-3 Groups of patients underwent urine diversion by percutaneous puncture nephrostomy, open nephrostomy and internal stent placement. Patients in Group 4 not underwent surgery with urine diversion. It was found that the long-term etiopathogenetic therapy against the background of retention changes resulted in progression of changes and loss of renal function in 63% of cases. Early urine diversion, depending on its method allows preserving the functional ability of the kidneys in 70.8 to 94.4% of cases. The combination of early renal drainage with antituberculous polychemotherapy is significantly superior to conservative treatment, leads to a rapid reduction of chronic renal failure (CRF) and allows to subsequently perform the greatest number of reconstructive operations: in 62.6% of cases after the external drainage and in 73.3%--after internal drainage (p < 0.05). At the same time, good results of plastics were achieved only in patients undergoing a two-stage surgical treatment, and poor results (relapse of stricture, progression of hydroureteronephrosis or CRF) were significantly more often observed (60%) in patients without urine diversion (p < 0.05).


Evaluation of pregnancy mortality in Louisiana using enhanced linkage and different indicators defined by WHO and CDC/ACOG: challenging and practical issues.
Differences in definitions and methods of data collection on deaths occurring during or shortly after pregnancy have created confusion and challenges in evaluating research findings. The study aimed to determine if the use of enhanced linkage procedures improve data collection of deaths occurring during or shortly after pregnancy, and how different definitions of those deaths changed the results of data analysis. The study used 2000-2005 Louisiana Pregnancy Mortality Surveillance System (LPMSS) and 2000-2005 death certificates linked with 1999-2005 live birth and fetal death certificates. Five indicators of deaths occurring during or shortly after pregnancy using WHO and CDC/ACOG definitions were estimated. One-sided Spearman rank test was used to analyze maternal mortality trends from 2000 to 2005. Of 345 women who died within 1 year of pregnancy, 187 were identified through linkage; 38 of those were missed by the LPMSS. Total mortality ratios of deaths occurring within 1 year of pregnancy ranged from 13.4 to 88.9 per 100,000 live births depending on the indicator used. CDC/ACOG pregnancy-related death and pregnancy-associated death statistically increased, whereas WHO pregnancy-related death decreased between 2000 and 2005. The most common causes of death differed by indicator. Universal adoption of linkage procedures could improve data on deaths occurring during or shortly after pregnancy. Estimates, trends, and most common causes of death were markedly different depending on which indicator was used. Additionally, the use of different mortality indicators during analysis provides a more detailed picture of potential target areas for future research and interventions.


Effects of interferon-alpha, verapamil and dacarbazine in the treatment of advanced malignant melanoma.
Treatment of patients with metastatic melanoma with either dacarbazine (DTIC) or interferon-alpha (IFNalpha) as single drugs, or in combination, results in a response rate of approximately 15-20%. This study evaluated the activity and toxicity following treatment with a combination of DTIC, IFNalpha2b and verapamil (VPL). Thirty patients with disseminated metastatic melanoma received DTIC 250 mg/m(2) on days 1-5 of a 4 week schedule, IFNalpha2b 3 MIU on days 1-5 each week, and VPL 80 mg three times a day throughout the cycle, until either disease progression or serious toxicity was observed. Among the 28 evaluable patients, there were four complete responses (CRs), five partial responses (PRs) and eight patients with stable disease (SD). The overall response rate (CR + PR) was 32%. Two patients with a CR were long-term survivors (45 and 34 months) and a third is still in complete remission after 49 months. The fourth CR patient relapsed and died with progressive brain metastases after 8 months. Among the eight patients with SD, one survived for 22 months and another for 34 months. Despite one toxic death, these results suggest that this treatment regimen is well tolerated and seems to be more effective than DTIC alone in a subset of patients. A controlled randomized study would be required to determine the value of adding VPL and IFNalpha2b to DTIC.


Wrist and hand ganglion treatment with hyaluronidase injection and fine needle aspiration: a tropical African perspective.
In an attempt to provide an alternative non-invasive treatment to surgical excision of ganglion cysts of the hand, and as part of the departmental audit resulting from the prevailing economic depression, 340 consecutive patients with 349 ganglia were treated in a prospective investigation by intralesional injection of hyaluronidase (up to 150 units in 1 ml) followed by fine needle aspiration (FNA) of the cyst to dryness. Pressure was applied over a piece of gauze and maintained with a crepe bandage for 24 h. Of the 340 patients treated in this way, the vast majority (323 or 95.0%) were considered to be cured on clinical examination at 6-month follow-up; only 17 patients (5.0%) exhibited recurrence during this period and these were successfully treated by re-aspiration. To the knowledge of this author, this is the first report of the use of the enzyme hyaluronidase as an adjunct to FNA in the treatment of ganglion cysts of the hand. The results clearly show that this method of treatment is a safe, fast, well accepted and cost-effective alternative to surgical excision, which is relatively expensive and is known to be associated with certain complications, including hypertrophic scars and cheloids.


The congenital undescended scapula. Surgical correction by the woodward procedure.
At The Hospital for Sick Children in Toronto, 21 undescended scapulae were corrected by the Woodward procedure in 20 patients over the past 18 years. The average age at operation was six years six months. The average follow-up period after operation was 8 years 9 months. Fifteen patients were girls and five were boys. All the patients had other associated abnormalities. Only one patient had a concomitant clavicular osteotomy. The average increase in glenohumeral abduction was 37 degrees. The average scapular lowering was two centimetres. Excellent or good cosmetic results were obtained in 80 per cent of patients. Ninety-three per cent of patients expressed satisfaction with the operative results. Complications were few. The Woodward procedure is a safe and relatively straightforward surgical means to effect a good cosmetic and functional improvement in a patient with a congenital undescended scapula. We do not recommend clavicular osteotomy as a routine, but it may be added to gain more correction with less risk of neurovascular compression.


Helichrysum italicum (Roth) G. Don and Helichrysum arenarium (L.) Moench Infusion Consumption Affects the Inflammatory Status and the Composition of Human Gut Microbiota in Patients with Traits of Metabolic Syndrome: A Randomized Comparative Study.
Helichrysum italicum (Roth) G. Don (HI) and Helichrysum arenarium (L.) Moench (HA) are rich in polyphenols and their infusions have beneficial effects for patients with metabolic syndrome. To investigate whether these effects are mediated by the gut microbiota, we analysed the effects of daily consumption of HI or HA infusion on the composition of gut microbiota, inflammatory status, and zonulin, a marker of gut barrier permeability. The study was a randomized, double-blind comparative trial. Thirty participants were randomly assigned to two groups and received either HA or HI tea filter bags, each containing 1 g of dried plant material, for daily consumption lasting 4 weeks. The results show that consumption of both infusions resulted in a reduction of some genera belonging to Firmicutes and in a slight but significant reduction in Shannon diversity index. Consumption of HI infusion significantly reduced serum levels of proinflammatory markers and zonulin alongside with the observed trend of Proteobacteria reduction. It can therefore be concluded that the HI and HA infusions could act as prebiotics and thus improve the intestinal environment. In addition, HI infusion has a positive impact on microbial dysbiosis and gut barrier dysfunction that occur in obesity and metabolic syndrome.


Effects of cimetidine and carbenoxolone on gastric mucus.
The effect of cimetidine on normal human gastric mucus has been compared with that of carbenoxolone, a drug believed to enhance mucus production. Each drug was given for two weeks, the gastric contents aspirated over a timed period and the results assessed in unstimulated and pentagastrin stimulated secretions. The volume, dry weight and the carbohydrate contents of non-diffusable glycoconjugates, high molecular mass glycoproteins and glycopolypeptides were investigated. Both drugs reduced the volumes of stimulated secretions. This was statistically significant after cimetidine. More importantly neither drug affected the amount of non-diffusable glycoconjugates, so that the concentration remained the same or increased. Both drugs reduced the monosaccharide content of the high molecular mass fractions. This reached significance for the stimulated secretion after cimetidine. As the carbohydrate content of the glycopolypeptides was unchanged this indicated the presence of a non-mucin glycoprotein or protein. Overall there was no fundamental difference between the results for cimetidine and carbenoxolone.


Development and application of a tool to assess laboratory hygiene in contained-use facilities.
To gain information on laboratory hygiene in contained-use laboratories, a method was developed to study the presence of microorganisms on laboratory equipment. Focusing detection on genetically modified organisms (GMOs) containing the universal M13 primer binding sites enabled the detection of a broad range of GMOs using a single PCR. Swabbing surfaces in three different contained-use laboratories led to detection of M13-containing PCR products in 26 out of 34 swabs. Most sequences (up to five per sample) were detected in swabs from the centrifuge and sink, followed by swabs taken from the bin and incubator (up to four sequences per sample). The obtained sequences varied in length from 171 nucleotides (nt) to 878 nt. In most cases, sequences were only partially similar to sequences published in GenBank. The lengths of the regions with high similarity varied from 94 nt to 795 nt, and these similarities ranged from 81% to 100%. Similarities with more than one sequence were commonly found, complicating the identification of detected sequences. Nonetheless, 84% of the detected sequences were actually handled in the laboratory at the time of sampling. This demonstrates that the method may be used as a quality control tool to assess the efficacy of decontamination and cleaning of commonly used surfaces, such as laboratory benches, freezer doors, and centrifuge rotors, without prior knowledge of the identity or characteristics of the GMOs.


Percutaneous Coronary Intervention versus Coronary Artery Bypass Grafting in Complex Coronary Artery Disease: Long-term Clinical Outcomes from a High-volume Center.
Clinical equipoise between a percutaneous coronary intervention (PCI) and coronary artery bypass grafting surgery (CABG) in the treatment of complex coronary artery disease (CAD), including unprotected left main coronary artery (LMCA) and/or three-vessel disease (3VD), remains debatable. A retrospective analysis of an unselected cohort undergoing contemporary PCI versus CABG at a large center in 2015. Patients who received nonemergent treatment of unprotected LMCA and/or 3VD were included. The primary study endpoint was all-cause mortality at 5 years. Secondary endpoints included a composite of all-cause mortality, spontaneous myocardial infarction (MI), or ischemia-driven repeat revascularization at 30 days and 1 year. Four hundred and thirty patients met the inclusion criteria, 225 had PCI, and 205 had CABG. PCI patients were older with frequent LMCA involvement and higher EuroSCORE yet they had a fourfold shorter in-hospital stay compared to CABG patients. At 5 years, there was no significant difference in the primary endpoint between CABG and PCI (adjusted Hazard ratios 0.68, 95% confidence interval: 0.38-1.22, P = 0.19). Likewise, there was no significant difference in the incidence of the secondary composite endpoint or its components at 30 days or 1 year. A propensity score-matched analysis in 220 patients revealed similar outcomes. In real-world long-term contemporary data, survival after PCI was comparable to CABG at 5 years in patients with unprotected LMCA and/or 3VD. At 1 year, the incidence of spontaneous MI and ischemia-driven repeat revascularization did not differ between the two cohorts. The mode of revascularization in these complex patients should be guided by the heart team.


Fractionation of transcription factors for RNA polymerase II from Drosophila Kc cell nuclear extracts.
Drosophila Kc cells were utilized to prepare nuclear extracts in which promoter-containing DNA templates were efficiently transcribed by RNA polymerase II. A combination of fractionation schemes was used to identify and partially purify seven activities (factors) which affected the transcription of four different genes in vitro. Reconstructing specific transcription required exogenous RNA polymerase II in addition to these factors. Moreover, the high efficiency of transcription characteristic of the crude extract was preserved in reconstruction reactions. The methods used are presented in detail. Functions were assigned to several of the factors. One essential factor appeared to affect initiation and displayed chromatographic properties unlike any other Drosophila transcription factor previously described. Two factors specifically affected RNA chain elongation. Another activity was a DNase inhibitor required to preserve template integrity in the fractionated system. The remaining three factors were not absolutely essential but affected the specific in vitro transcription either qualitatively or quantitatively. A comparison of these transcription factors with other Drosophila and mammalian transcription factors is made.


Serum levels of the soluble adhesion molecules in patients with malignant melanoma.
The incidence of malignant melanoma has been steadily increasing over the past decades. CD 44 is a transmembrane glycoprotein which is implicated in a number of adhesive and migratory events. Downregulation of CD 44 is implicated in the metastatic process. P-Selectin is a member of the selectin family of cell surface molecules. The levels of P-Selectin in biological fluids may be elevated in subjects with a variety of pathological conditions. In malignant melanoma, elevation of the plasma level of soluble intercellular adhesion molecule-1 (sICAM-1) has been associated with a reduction in disease-free survival. This study was performed to investigate the differences in the serum concentrations of the adhesion molecules in patients with malignant melanoma. The study group consisted of 52 patients with malignant melanoma and 20 healthy subjects. No meaningful difference was observed for P-selectin and sICAM 1 levels. A statistically significant decrease was observed in the cancer patients for serum CD 44 levels.


Ixodes ventalloi: morphological and molecular support for species integrity.
Despite their medical and veterinary importance, some tick species are so poorly studied, that their role within pathogen vector transmission cycles is difficult to assess. The tick Ixodes ventalloi is one such species, and its biology and phylogenetic status remain an issue of debate. In the present study, specimens of adult I. ventalloi (n = 65 females; n = 31 males) infesting cats in the Lipari Island (Aeolian archipelago, Sicily, southern Italy) were characterized morphologically and molecularly, the latter based on mitochondrial 16S rRNA and cytochrome c oxidase subunit 1 (cox1) genes. The genetic data and phylogenetic analyses for both mitochondrial genes suggest the existence of two distinct genogroups. The ecological and epidemiological significance of the genetic structure within the I. ventalloi endemic population remains to be determined. The results highlight the need for further analysis of this tick species, including whole mitochondrial genome sequencing and crossbreeding studies, which will be pivotal to complement features of its status as a vector of pathogens.


Validating TRANSP simulations using neutron emission spectroscopy with dual sight lines.
A method to generate modeled neutron spectra from bulk and fast ion distributions simulated by TRANSP has been developed. In this paper, modeled data generated from fuel ion distributions modeled with TRANSP is compared to measured data from two neutron spectrometers with different lines of sight; TOFOR with a radial one and the MPRu with a tangential one. The information obtained from the analysis of the measured neutron spectra such as the relative intensity of the emission from different ion populations places additional constraints on the simulation and can be used to adjust the parameters of the simulation.


Attenuation of the effects of rat hemorrhagic shock with a reperfusion injury-inhibiting agent specific to mice.
Death after hemorrhagic shock (HS) may be caused by a generalized reperfusion injury, particularly noticeable in the gut. A period of tissue ischemia followed by reinstitution of perfusion produces severe inflammation that can be blocked in mice by preventing the binding of a pathogenic natural immunoglobulin M (IgM) of defined specificity to antigens in reperfused tissue by using a soluble peptide analogue of the IgM tissue target. We hypothesize that this agent can improve end points of rat HS: death, intestinal injury, and lung injury. Male Sprague-Dawley rats were anesthetized; 50% of calculated blood volume was removed for 120 min, shed blood, then returned; and animals were sacrificed at 72 h. One group of rats received i.v. analogue ([N2] 300 microg) with the return of shed blood. Small intestine and lung were evaluated by histological examination and immunohistochemistry. Lung edema was assessed by Evans blue extravasation and histological examination. I.v. N2 decreased experimental mortality from 62% to 12% (P < 0.05). Associated with this was diminution of gut injury score from 57.8% +/- 5.5% to 19.5% +/- 2.5% (P < 0.05), lung injury from 21.4 +/- 1.5 to 14.8 +/- 1.3 polymorphonuclear leucocytes per high-power field x400 (P < 0.05), and Evans blue extravasation index from 0.61 +/- 0.14 to 0.18 +/- 0.06 (P < 0.05). As well, the deposition of IgM and C3 that is seen in intestinal villi from HS was not present in N2-treated rats. The N2 peptide agent that blocks reperfusion injury in mice prevents death from rat HS, as well as attenuates gut reperfusion injury and its remote target injuries. These data suggest that death from HS is caused by reperfusion injury, and that an agent derived from mice is effective in rats when given in real therapeutic time.


Long noncoding RNA LOC646029 functions as a ceRNA to suppress ovarian cancer progression through the miR-627-3p/SPRED1 axis.
Long noncoding RNAs (lncRNAs) play a crucial regulatory role in the development and progression of multiple cancers. However, the potential mechanism by which lncRNAs affect the recurrence and metastasis of ovarian cancer remains unclear. In the current study, the lncRNA LOC646029 was markedly downregulated in metastatic ovarian tumors compared with primary tumors. Gain- and loss-of-function assays demonstrated that LOC646029 inhibits the proliferation, invasiveness, and metastasis of ovarian cancer cells in vivo and in vitro. Moreover, the downregulation of LOC646029 in metastatic ovarian tumors was strongly correlated with poor prognosis. Mechanistically, LOC646029 served as a miR-627-3p sponge to promote the expression of Sprouty-related EVH1 domain-containing protein 1, which is necessary for suppressing tumor metastasis and inhibiting KRAS signaling. Collectively, our results demonstrated that LOC646029 is involved in the progression and metastasis of ovarian cancer, which may be a potential prognostic biomarker.


Developmental regulation of bicoid mRNA stability is mediated by the first 43 nucleotides of the 3' untranslated region.
During the transition from the maternal to the zygotic developmental program, the expression of genes important for pattern formation or cell cycle regulation changes dramatically. Rapid changes in gene expression are achieved in part through the control of mRNA stability. This report focuses on bicoid, a gene essential for formation of anterior embryonic structures in Drosophila melanogaster. bicoid mRNA is synthesized exclusively during oogenesis. Here, we show that bicoid mRNA stability is regulated. While bicoid mRNA is stable in retained oocytes, in unfertilized eggs, and during the first 2 h of embryogenesis, specific degradation is activated at cellularization of the blastoderm. To identify cis-acting sequences required for bicoid mRNA's regulated stability, fusions between bicoid and genes producing stable mRNAs were introduced into the Drosophila germ line by P-element-mediated transformation. The analysis of the fusion mRNAs identified a bicoid instability element (BIE) contained within a 43-nucleotide sequence immediately following the stop codon. The BIE is sufficient to destabilize the otherwise-stable ribosomal protein A1 mRNA and is separable from the previously identified bicoid mRNA localization signals and from the "nanos response element." Similar mechanisms may regulate a class of developmentally important maternal genes whose mRNA has a temporal profile similar to that of bicoid.


Profiling of immune-cancer interactions at the single-cell level using a microfluidic well array.
Cancer immunotherapy has achieved great success in hematological cancers. However, immune cells are a highly heterogeneous population and can vary highly in clonal expansion, migration and function status, making it difficult to evaluate and predict patient response to immune therapy. Conventional technologies only yield information on the average population information of the treatment, masking the heterogeneity of the individual T cell activation status, the formation of immune synapse, as well as the efficacy of tumor cell killing at the single-cell level. To fully interrogate these single-cell events in detail, herein, we present a microfluidic microwell array device that enables the massive parallel analysis of the immunocyte's heterogeneity upon its interaction pairs with tumor cells at the single-cell level. By precisely controlling the number and ratio of tumor cells and T cells, our technique can interrogate the dynamics of the CD8+ T cell and leukemia cell interaction inside 6400 microfluidic wells simultaneously. We have demonstrated that by investigating the interactions of T cell and tumor cell pairs at the single-cell level using our microfluidic chip, details hidden in bulk investigations, such as heterogeneity in T cell killing capacity, time-dependent killing dynamics, as well as drug treatment-induced dynamic shifts, can be revealed. This method opens up avenues to investigate the efficacy of cancer immunotherapy and resistance at the single-cell level and can explore our understanding of fundamental cancer immunity as well as determine cancer immunotherapy efficacy for personalized therapy.


The Effects of Machine-Weight and Free-Weight Resistance Exercise on Hemodynamics and Vascular Function.
The purpose of this study was to examine hemodynamic and vascular responses between machine-weight and free-weight exercise. Resistance-trained individuals were assigned to a machine-weight (n = 13) or free-weight (n = 15) group. Groups completed two visits consisting of their assigned exercise condition and a control (CON). A 2 × 2 × 3 repeated measures ANOVA was used to test the effects of group across condition and time on the hemodynamic parameters [cardiac output (CO), heart rate (HR), total peripheral resistance (TPR), mean arterial pressure (MAP), and stroke volume (SV)]. A 2 × 2 × 2 repeated measures ANOVA was used to test the effects of group across condition and time on the hemodynamic variable, forearm vascular conductance (FVC), as well as on vascular measures [forearm blood flow (FBF), blood flow peak, and total reactive hyperemia (RH)]. Main effects were analyzed using pairwise comparisons. The results of the present study demonstrate that both machine-weight and free-weight exercise produce similar (p > 0.05) alterations in hemodynamics and vascular function. Specifically, during recovery both groups demonstrated significant (p ≤ 0.05) increases in measures of hemodynamics such as CO, HR and FVC, as well as significant (p ≤ 0.05) decreases in TPR, MAP, and SV. Measures of vascular function such as FBF, blood flow peak, and total RH were also significantly (p ≤ 0.05) increased during recovery. Therefore, this study suggests that either machine weight or free-weight exercise may induce acute hemodynamic and vascular benefits, which may reduce the risk of cardiovascular disease (CVD) and CVD events.


Effects of NOD-like receptors in human B lymphocytes and crosstalk between NOD1/NOD2 and Toll-like receptors.
NLRs are recently discovered PRRs detecting substructures of peptidoglycans and triggering innate immunity. NLRs are expressed in several cell types, but the presence in human B lymphocytes is still unknown. This study aimed to investigate expression and function of NLRs in human B lymphocytes. B cells were isolated and analyzed for mRNA and protein expression. The functional responsiveness of NOD1 and NOD2 was investigated upon stimulation with the cognate ligands, with or without stimulation via IgM/IgD/CD40 and/or selected TLR agonists. A differential expression of NLRs was demonstrated in blood-derived and tonsillar B cells, whereas no variations were found among naive, germinal center, or memory B cells. Stimulation with the ligands alone did not induce B cell activation. However, upon concomitant BCR triggering, an increase in proliferation was seen, together with an induction of cell surface markers (CD27, CD69, CD71, CD80, CD86, and CD95) and prolonged survival. Peripheral B cells were activated by NOD1 and NOD2 ligands, whereas tonsil-derived B cells responded solely to NOD1. In contrast, costimulation with CD40L failed to induce activation. Additionally, it was found that NLR ligands could enhance TLR-induced proliferation of B cells. The present study demonstrates expression of functional NLRs in human B cells. We show that NOD1 and NOD2 have the ability to augment the BCR-induced activation independently of physical T cell help. Hence, NLRs represent a new pathway for B cell activation and a potentially important host defense system against bacterial infections.


Hepatitis A virus (HAV) proteinase 3C inhibits HAV IRES-dependent translation and cleaves the polypyrimidine tract-binding protein.
Hepatitis A virus (HAV) infection is still an important issue worldwide. A distinct set of viruses encode proteins that enhance viral cap-independent translation initiation driven by an internal ribosome entry site (IRES) and suppress cap-dependent host translation. Unlike cytolytic picornaviruses, replication of HAV does not cause host cell shut off, and it has been questioned whether HAV proteins interfere with its own and/or host translation. HAV proteins were coexpressed in Huh-7 cells with reporter genes whose translation was initiated by either cap-dependent or cap-independent mechanisms. Among the proteins tested, HAV proteinase 3C suppressed viral IRES-dependent translation. Furthermore, 3C cleaved the polypyrimidine tract-binding protein (PTB) whose interaction with the HAV IRES had been demonstrated previously. The combined results suggest that 3C-mediated cleavage of PTB might be involved in down-regulation of viral translation to give way to subsequent viral genome replication.


Computer-aided design and computer-assisted manufacturing in prosthetic implant dentistry.
The aim of this systematic review was to evaluate the existing scientific evidence on human clinical studies describing the application of computer-aided design/computer-assisted manufacturing (CAD/CAM) technology in restorative implant dentistry. Electronic searches for clinical studies from 1966 through May 2008 focusing on long-term follow-up were performed using the PubMed search engine. Concentrating on the restorative aspect of the CAD/CAM technology applicable to implant dentistry, pertinent literature was divided into articles related to implant abutments and implant frameworks. Of the 885 articles initially reviewed, 5 articles (3 CAD/CAM framework and 2 CAD/CAM abutment) satisfied the search criteria of the literature search performed. Combining the results from the framework clinical trial studies, there were a total of 189 prostheses supported by 888 implants. The follow-up varied between 12 and 60 months. Four implants were lost prior to the insertion of the prosthesis and 46 after the insertion. One prosthesis failure was reported. Similarly, in the 2 abutment clinical trial studies there were a total of 53 ceramic abutments supported by 53 implants. The patients were followed between 12 and 44 months. No significant failures or complications were reported in association with the implants and their restorations. Based on a systematic review of literature concerning CAD/CAM used for fabrication of frameworks and abutments, preliminary proof of concept was established. Clinical studies on the use of these techniques were too preliminary and underpowered to provide meaningful conclusions regarding the performance of these abutments/frameworks.