Conformationally constrained analogues of diacylglycerol (DAG). 14. Dissection of the roles of the sn-1 and sn-2 carbonyls in DAG mimetics by isopharmacophore replacement.
The replacement of the sn-1 and sn-2 carbonyl esters in DAG-surrogate lactones by sulfonate esters showed that their isosteric properties in protein kinase C binding are controlled by the location of the hydrophobic alkyl chain on the molecule. The CO and SO2 groups appear to be true isosteres only when they are adjacent to the alkyl chain, which is presumed to insert normal to the lipid bilayer.


Deep learning methods for medical image fusion: A review.
The image fusion methods based on deep learning has become a research hotspot in the field of computer vision in recent years. This paper reviews these methods from five aspects: Firstly, the principle and advantages of image fusion methods based on deep learning are expounded; Secondly, the image fusion methods are summarized in two aspects: End-to-End and Non-End-to-End, according to the different tasks of deep learning in the feature processing stage, the non-end-to-end image fusion methods are divided into two categories: deep learning for decision mapping and deep learning for feature extraction. According to the different types of the networks, the end-to-end image fusion methods are divided into three categories: image fusion methods based on Convolutional Neural Network, Generative Adversarial Network, and Encoder-Decoder Network; Thirdly, the application of the image fusion methods based on deep learning in medical image field is summarized from two aspects: method and data set; Fourthly, evaluation metrics commonly used in the field of medical image fusion are sorted out from 14 aspects; Fifthly, the main challenges faced by the medical image fusion are discussed from two aspects: data sets and fusion methods. And the future development direction is prospected. This paper systematically summarizes the image fusion methods based on the deep learning, which has a positive guiding significance for the in-depth study of multi modal medical images.


A simple preparation step to remove excess liquid lipids in white adipose tissue enabling improved detection of metabolites via MALDI-FTICR imaging MS.
Matrix-assisted laser desorption ionization (MALDI) Fourier transform ion cyclotron resonance (FTICR) imaging mass spectrometry (MS) is a powerful technology used to analyze metabolites in various tissues. However, it faces significant challenges in studying adipose tissues. Poor matrix distribution and crystallization caused by excess liquid lipids on the surface of tissue sections hamper m/z species detection, an adverse effect that particularly presents in lipid-rich white adipose tissue (WAT). In this study, we integrated a simple and low-cost preparation step into the existing MALDI-FTICR imaging MS pipeline. The new method-referred to as filter paper application-is characterized by an easy sample handling and high reproducibility. The aforementioned filter paper is placed onto the tissue prior to matrix application in order to remove the layer of excess liquid lipids. Consequently, MALDI-FTICR imaging MS detection was significantly improved, resulting in a higher number of detected m/z species and higher ion intensities. After analyzing various durations of filter paper application, 30 s was found to be optimal, resulting in the detection of more than 3700 m/z species. Apart from the most common lipids found in WAT, other molecules involved in various metabolic pathways were detected, including nucleotides, carbohydrates, and amino acids. Our study is the first to propose a solution to a specific limitation of MALDI-FTICR imaging MS in investigating lipid-rich WAT. The filter paper approach can be performed quickly and is particularly effective for achieving uniform matrix distribution on fresh frozen WAT while maintaining tissue integrity. It thus helps to gain insight into the metabolism in WAT.


Tetracyclic triterpenes. Part VI [1]. The synthesis of 4,4,14 alpha-trimethyl-19 (10 leads to 9 beta) abeo-steroids. Synthons for the preparation of cucurbitacins.
The effective synthesis of 4,4,14 alpha-trimethyl-19 (10 leads to 9 beta) abeo-steroids (IV), (V), and (VI) with two- and five-carbon side chains from lanosterol is described. Their structures were proved on the basis of spectral data. The title compounds are the first synthetic synthons for the preparation of 4,4,14 alpha-trimethyl-steroids with an unnatural configuration.


Meeting highlights of the 8th Annual Scientific Sessions of the Society For Cardiovascular Magnetic Resonance, January 21 to 23, 2005.
Parallel tracks for clinical scientists, basic scientists, and pediatric imagers was the novel approach taken for the highly successful 8th Annual Scientific Sessions of the Society for Cardiovascular Magnetic Resonance, held in San Francisco, California, January 21 to 23, 2005. Attendees were immersed in information on the latest scientific advances in cardiovascular magnetic resonance (CMR) from mice to man and technological advances from systems with field strengths from 0.5 T to 11.7 T. State-of-the-art applications were reviewed, spanning a wide range from molecular imaging to predicting outcome with CMR in large patient populations.


Increased indices of lipid peroxidation in stress-susceptible pigs and effects of vitamin E.
When stress-susceptible and stress-resistant pigs consumed diets containing 10 iu vitamin E kg-1, the stress-susceptible pigs had damaged cell membranes. This was indicated by increased plasma activities of creatine kinase and pyruvate kinase. Plasma concentrations of thiobarbituric acid reactive substances and conjugated dienes were also increased suggesting that the membrane damage was caused by free radicals. Susceptibility to free radicals was further demonstrated by an increased tendency for erythrocytes and tissue homogenates from stress-susceptible pigs to peroxidise when incubated with hydrogen peroxide and iron, respectively. Supplementation of the diets with approximately 20 times normal requirements of vitamin E decreased plasma creatine kinase and pyruvate kinase activities, and inhibited the formation of the indices of peroxidation in vivo and in vitro. It is concluded that the stress-susceptible pig has an abnormality in its antioxidant defence mechanisms. However, this abnormality was not due to a deficiency in selenium-dependent glutathione peroxidase, since activities were significantly increased in the longissimus dorsi of stress-susceptible pigs. Although the nature of the antioxidant defect is unclear, it is suggested that stress-susceptible pigs are under a sustained oxidant stress and that a decreased ability to accommodate even a normal free radical load may contribute to the rapid development of the fatal stress response.


Xanthogranulomatous cholecystitis: diagnostic complexity and review of the literature.
We report the case of a 39-year-old male presenting with acute onset vomiting and diarrhoea. Initially treated empirically for gastroenteritis, imaging later confirmed a complicated episode of cholecystitis with fistular formation and intra-abdominal cyst. Following cholecystectomy, histology confirmed a case of xanthogranulomatous cholecystitis (XGC). This paper presents a detailed summary of the condition alongside a literature review of all available episodes of XGC since 2017 with the aim of highlighting diagnostic conclusions regarding the nature of the disease and its clinical manifestations.


Genotyping and biofilm formation of Mycoplasma hyopneumoniae and their association with virulence.
Mycoplasma hyopneumoniae, the causative agent of swine respiratory disease, demonstrates differences in virulence. However, factors associated with this variation remain unknown. We herein evaluated the association between differences in virulence and genotypes as well as phenotype (i.e., biofilm formation ability). Strains 168 L, RM48, XLW-2, and J show low virulence and strains 232, 7448, 7422, 168, NJ, and LH show high virulence, as determined through animal challenge experiments, complemented with in vitro tracheal mucosa infection tests. These 10 strains with known virulence were then subjected to classification via multilocus sequence typing (MLST) with three housekeeping genes, P146-based genotyping, and multilocus variable-number tandem-repeat analysis (MLVA) of 13 loci. MLST and P146-based genotyping identified 168, 168 L, NJ, and RM48 as the same type and clustered them in a single branch. MLVA assigned a different sequence type to each strain. Simpson's index of diversity indicates a higher discriminatory ability for MLVA. However, no statistically significant correlation was found between genotypes and virulence. Furthermore, we investigated the correlation between virulence and biofilm formation ability. The strains showing high virulence demonstrate strong biofilm formation ability, while attenuated strains show low biofilm formation ability. Pearson correlation analysis revealed a significant positive correlation between biofilm formation ability and virulence. To conclude, there was no association between virulence and our genotyping data, but virulence was found to be significantly associated with the biofilm formation ability of M. hyopneumoniae.


Prevalence and risk factors of primary open-angle glaucoma in a city of Eastern China: a population-based study in Pudong New District, Shanghai.
The aim of the present study was to investigate the prevalence and characteristics of primary open-angle glaucoma among the urban population of Pudong New District, Shanghai. Three residents' committees were randomly selected from Pudong New District, and residents aged 50 and older were screened for primary open-angle glaucoma (POAG) from March to April 2011. In remote screening, the tests on visual acuity, refraction, intraocular pressure (IOP), and the photographs of anterior segment and fundus were used to identify POAG suspect. The suspected subjects were then reexamined with the tests on IOP, gonioscopy, Humphrey visual field test, and retinal nerve fiber layer (RNFL) thickness by optical coherence tomography (OCT). POAG was diagnosed according to the criteria defined by International Society for Geographic and Epidemiological Ophthalmology (ISGEO). Finally, POAG risk factors were evaluated using logistic regression analysis. A total of 2528 citizens out of 3,146 eligible residents (80.36 %) participated in this study. Among the citizens, 72 were diagnosed to have POAG, giving the crude prevalence rate of 2.85 % (95 % CI:2.20 %-3.50 %) in general and age- and gender-adjusted prevalence rate of 2.8 % (95 % CI: 2.78 %-2.83 %). Among the 72 POAG patients, only 22 cases had IOP exceeding 21 mmHg while other 50 cases had IOP of 21 mmHg or less; nine cases had one eye blind (12.5 %). Intriguingly, only eight cases (11.11 %) had been diagnosed with POAG before this screening. More efforts are required for early screening and education on POAG in communities, especially in a POAG high-risk population.


Single Treatment with 100-Microsecond Alexandrite Laser Clears Selected Acquired Melanocytic Nevi in Type IV Asian Facial Skin.
Small common acquired melanocytic nevi (AMNs) are common on Asian facial skin. To show that the 755 nm Alexandrite laser stacked at the 100-μs long-pulsed mode (μsAL) is an effective modality for the removal of selected AMNs. This was a retrospective case series, followed up with a telephone interview. A retrospective analysis of all patients treated between January 2010 and April 2012 with the μsAL laser for small AMNs was conducted. Pre- and post-treatment facial photographs and photographs of the individual lesions were analyzed by two independent dermatological surgeons for degree of clearance and complications. A telephone interview was conducted with the patients to assess their satisfaction with the procedure. A total of 18 patients with 53 lesions were included. 7/18 (38.9%) of patients had 'excellent' results. No patients had 'mild' or 'poor' results. At 4 week post-treatment, 49/53 (92.5%) were totally cleared, with 14/53 (26.4%) reporting mild atrophy, and 11/53 (20.8%) reporting mild post-inflammatory hypopigmentation. The majority of lesions had negligible complications. 9/18 (50%) judged the procedure to be 'excellent', and all patients reported that they would recommend this procedure to a friend seeking removal of small facial AMNs. The μsAL is an effective modality for the removal of small facial AMNs.


Ultrastructural organization of thrombocytes in normal monkeys (Macaca arctoides) and in those with experimental viral malignant lymphoma.
Data on platelet ultrastructure of 6 M. arctoides monkeys investigated before and during experimental viral malignant lymphoma are presented. It has been established that the inner structure of thrombocytes in healthy monkeys was almost similar to that of man. In experimental malignant lymphoma, irrespective of the clinical period of the disease, a decrease of alpha-, beta- and sigma-granules, a decrease of the functional activity of thrombocytes and an increase of delta-granules and glycogen granulation content were observed. The above mentioned disturbances can be a result of changes of megakaryocytes in malignant lymphoma. No viral particles in thrombocytes of neither healthy nor sick monkeys have been revealed.


Exercise increased BDNF and trkB in the contralateral hemisphere of the ischemic rat brain.
Previous studies have suggested that brain-derived neurotrophic factor (BDNF) and trkB both have a role in plasticity following brain insults and exercise increases BDNF and trkB mRNA levels in the normal brain. We attempted to determine whether treadmill exercise improves motor function following experimental cerebral ischemia, and whether motor outcome is associated with BDNF and trkB expression. We subjected adult male Sprague-Dawley rats to a permanent ischemia, followed by either 12 days of treadmill exercise or non-exercise. In the exercise group, improvements in the motor behavior index were found and BDNF and trkB proteins in contralateral hemisphere were increased. This study suggests that after permanent brain ischemia, exercise improves motor performance and elevates BDNF and trkB proteins in the contralateral hemisphere.


Depression in Mexican medical students: A path model analysis.
Depression is a mental health disorder present in medical students that foster cumulative disadvantages throughout life and that may compromise patient care. Yet, in Mexico there is a dearth of research concerning prediction models for depression in this population. The aim of this cross-sectional study was to develop and test a path model analysis of depression in 103 freshmen Mexican medical students ages 18-23 years old selected non-randomly. Anxiety, emotion dysregulation, and coping skills were used as factors influencing depression. Mexican self-report scales were used as assessment measures. Main results showed that anxiety and emotion dysregulation had a significant direct influence on depression. Emotion dysregulation had a significant direct effect on coping skills and anxiety, while anxiety had a significant direct effect on problem drinking and alcohol problems. Fit indexes obtained indicate an excellent adjustment to data. R square indicates that 53.7% of the variance in depression can be explained by this model. The results are exploratory rather than confirmatory. The sample size was not large enough and the research focused on only Mexican freshmen medical students. Other factors associated to depression in medical students were not considered. The current research provides unique findings in terms of a model of depression in Mexican medical students through which it is possible to design and implement interventions that decrease depression, thus positively affecting their well-being, their future careers as medical doctors and their abilities to serve the society adequately, especially in the context of Latin American cities.


Prospectively Reallocating Sedentary Time: Associations with Cardiometabolic Health.
This study aimed to investigate whether prospectively reallocating time away from sedentary behavior (SB) into different physical activity intensities is associated with 12-month change to cardiometabolic health in a cohort at high risk of type 2 diabetes (T2DM). Participants with known risk factors for T2DM were recruited from primary care (Leicestershire, United Kingdom) as part of the Walking Away from Type 2 Diabetes trial (n = 808). Participants were followed up at 12, 24, and 36 months. SB, light-intensity physical activity (LPA) and moderate-to-vigorous intensity physical activity (MVPA) were measured objectively by accelerometer. Postchallenge glucose, triglycerides, HDL cholesterol, systolic blood pressure, and waist circumference were analyzed individually and combined into a clustered cardiometabolic risk score (CMRS). Associations of changing SB over each consecutive 12-month period were analyzed taking account of repeated measures. Reallocating 30 min from SB to LPA was associated with 0.21-cm (95% confidence interval, 0.03-0.38 cm) reduction in waist circumference, 0.09-mmol·L (0.04-0.13 mmol·L) reduction in 2-h glucose, 0.02-mmol·L (0.00-0.04 mmol·L) reduction in triglycerides, and 0.02 (0.01-0.03) reduction in CMRS. Every 30-min reallocation from SB to MVPA was associated with 1.23-cm (0.68-1.79 cm) reduction in waist circumference, 0.23-mmol·L (0.10-0.36 mmol·L) reduction in 2-h glucose, 0.04-mmol·L (0.00-0.09 mmol·L) reduction in triglycerides, and 0.07 (0.04-0.11) reduction in CMRS. Reallocating 30 min from LPA into MVPA was also associated with 1.02-cm (0.43-1.60 cm) reduction in waist circumference, 0.16-mmol·L (0.02-0.30 mmol·L) reduction in 2-h glucose, and 0.05 (0.01-0.09) reduction in CMRS. Over 12 months, reallocating time away from SB into LPA or MVPA was associated with improved cardiometabolic health in a population at risk of T2DM, with the greatest benefits observed for MVPA.


Effect of three endodontic sealers on the bond strength of prefabricated fiber posts luted with three resin cements.
There is limited information in the literature regarding the effect of eugenol-based sealers on the bond strength of resin-bonded endodontic posts. The purpose of this study was to evaluate the effect of 1 resin-based and 2 different eugenol-based endodontic sealers on the bond strength of prefabricated fiber posts luted with 3 different resin cements. One hundred thirty-five prefabricated fiber posts were luted into extracted single-rooted teeth with 1 of 3 composite resin cements (Rely X Unicem, Paracore, and Variolink II). Specimens were divided into 3 groups with 45 teeth each. The first 2 groups were obturated with gutta percha and 1 of 2 eugenol-based endodontic sealers (Endofil, Tubli-Seal). The third group was obturated with a resin-based root canal sealer (AH26). The forces required for dislodgment of posts from their prepared post spaces were recorded by using a universal testing machine. Data were collected and a 2-way ANOVA was applied to the mean retentive strengths of various combinations of sealer and cement. A Tukey multiple comparison test was performed to determine which groups were significantly different (α=.05). Endofil and Tubli-Seal (eugenol-based sealers) groups had significantly lower bond strengths for the posts than the AH26 group (P<.001). There was no significant difference between the means of post bond strength for the Endofil and Tubli-Seal groups. There was significant difference among the means of post bond strength for all 3 cement groups (P<.05). Endofil and Tubli-Seal (eugenol-based) sealers demonstrated significantly reduced mean bond strength for prefabricated fiber posts luted with resin cement. Rely X Unicem resin cement exhibited higher mean bond strength than with Paracore and Variolink II resin cements when a eugenol sealer was used.


Marketing Is Not a Luxury - Marketing Is a Necessity.
Physicians who work in small practices constantly face a basic marketing question: What's the best way to sell myself and my business to the community? Because few physicians studied business principles, many won't know the answer. That's why the Texas Medical Association is publishing a book titled Marketing Smart: A Guide for Medical Practices.


Modified Stancu operators based on inverse Polya Eggenberger distribution.
In this paper, we construct a sequence of modified Stancu-Baskakov operators for a real valued function bounded on [Formula: see text], based on a function [Formula: see text]. This function [Formula: see text] is infinite times continuously differentiable on [Formula: see text] and satisfy the conditions [Formula: see text] and [Formula: see text] is bounded for all [Formula: see text]. We study the degree of approximation of these operators by means of the Peetre K-functional and the Ditzian-Totik modulus of smoothness. The quantitative Voronovskaja-type theorems are also established in terms of the first order Ditzian-Totik modulus of smoothness.


Rupture of sinus of Valsalva aneurysm: a case report in a child.
Sinus of Valsalva aneurysm (SVA) is a rare congenital disease that can cause severe clinical presentations when the aneurysm ruptures. Here, we report a rare case of a noncoronary sinus of Valsalva aneurysm with rupture into the right atrium. A 14-year-old Chinese female patient presented viral myocarditis with acute heart failure at the local hospital, and she was finally diagnosed with a noncoronary sinus Valsalva aneurysm with rupture into the right atrium by digital subtraction angiography with cardiac catheterization angiography and echocardiography at our hospital (Children's Hospital of Chongqing Medical University). Percutaneous closure intervention was performed shortly after her diagnosis, and the patient showed good functional recovery. We report a case of ruptured sinus of Valsalva aneurysm successfully treated by percutaneous closure, which is an excellent alternative treatment.


In vivo relevance for platelet glycoprotein Ibalpha residue Tyr276 in thrombus formation.
Platelet glycoprotein (GP) Ib-IX-V supports platelet adhesion on damaged vascular walls by binding to von Willebrand factor (VWF). For several decades it has been recognized that the alpha-subunit of GP (GPIbalpha) also binds thrombin but the physiological relevance, if any, of this interaction was unknown. Previous studies have shown that a sulfated tyrosine 276 (Tyr276) is essential for thrombin binding to GPIbalpha. This study investigated the in vivo relevance of GPIbalpha residue Tyr276 in hemostasis and thrombosis. Transgenic mouse colonies expressing the normal human GPIbalpha subunit or a mutant human GPIbalpha containing a Phe substitution for Tyr276 (hTg(Y276F)) were generated. Both colonies were bred to mice devoid of murine GPIbalpha. Surface-expressed GPIbalpha levels and platelet counts were similar in both colonies. hTg(Y276F) platelets were significantly impaired in binding alpha-thrombin but displayed normal binding to type I fibrillar collagen and human VWF in the presence of ristocetin. In vivo thrombus formation as a result of chemical damage (FeCl(3)) demonstrated that hTg(Y276F) mice have a delayed time to occlusion followed by unstable blood flow indicative of embolization. In models of laser-induced injury, thrombi developing in hTg(Y276F) animals were also less stable. The results demonstrate that GPIbalpha residue Tyr276 is physiologically important, supporting stable thrombus formation in vivo.


Deoxyelephantopin-a novel PPARγ agonist regresses pressure overload-induced cardiac fibrosis via IL-6/STAT-3 pathway in crosstalk with PKCδ.
Pathological cardiac hypertrophy is associated with ventricular fibrosis leading to heart failure. The use of thiazolidinediones as Peroxisome Proliferator-Activated Receptor-gamma (PPARγ)-modulating anti-hypertrophic therapeutics has been restricted due to major side-effects. The present study aims to evaluate the anti-fibrotic potential of a novel PPARγ agonist, deoxyelephantopin (DEP) in cardiac hypertrophy. AngiotensinII treatment in vitro and renal artery ligation in vivo were performed to mimic pressure overload-induced cardiac hypertrophy. Myocardial fibrosis was evaluated by Masson's trichrome staining and hydroxyproline assay. Our results showed that DEP treatment significantly improves the echocardiographic parameters by ameliorating ventricular fibrosis without any bystander damage to other major organs. Following molecular docking, all-atomistic molecular dynamics simulation, reverse transcription-polymerase chain reaction and immunoblot analyses, we established DEP as a PPARγ agonist stably interacting with the ligand-binding domain of PPARγ. DEP specifically downregulated the Signal Transducer and Activator of Transcription (STAT)-3-mediated collagen gene expression in a PPARγ-dependent manner, as confirmed by PPARγ silencing and site-directed mutagenesis of DEP-interacting PPARγ residues. Although DEP impaired STAT-3 activation, it did not have any effect on the upstream Interleukin (IL)-6 level implying possible crosstalk of the IL-6/STAT-3 axis with other signaling mediators. Mechanistically, DEP increased the binding of PPARγ with Protein Kinase C-delta (PKCδ) which impeded the membrane translocation and activation of PKCδ, downregulating STAT-3 phosphorylation and resultant fibrosis. This study, therefore, for the first time demonstrates DEP as a novel cardioprotective PPARγ agonist. The therapeutic potential of DEP as an anti-fibrotic remedy can be exploited against hypertrophic heart failure in the future.


Neuronal SH2B1 attenuates apoptosis in an MPTP mouse model of Parkinson's disease via promoting PLIN4 degradation.
The incidence of Parkinson's disease (PD) has increased tremendously, especially in the aged population and people with metabolic dysfunction; however, its underlying molecular mechanisms remain unclear. SH2B1, an intracellular adaptor protein, contributes to the signal transduction of several receptor tyrosine kinases and exerts beneficial metabolic effects for body weight regulation; however, whether SH2B1 plays a major role in pathological neurodegeneration in PD has not yet been investigated. This study aimed to investigate the effects of SH2B1 in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mice with Sh2b1 deficiency or neuron-specific Sh2b1 overexpression. Cellular and molecular mechanisms were elucidated using human dopaminergic neuron SH-SY5Y cells analysed. We found that SH2B1 expression was confirmed to be downregulated in the blood samples of PD patients and in the brains of mice with MPTP-induced chronic PD. Sh2b1 deficiency caused marked exacerbation of behavioural defects and increased neuronal apoptosis in MPTP-treated mice, whereas restoration of neuron-specific Sh2b1 expression significantly reversed these effects. Similar results were observed in MPP + -treated SH-SY5Y cells. Mechanistically, upon binding to heat shock cognate 70 (HSC70), SH2B1 promotes HSC70-related recognition and PLIN4 lysosomal translocation and degradation, thus suppressing lipid peroxidation stress in the brains of PD mice. Adeno-associated virus-mediated rescue of neuronal HSC70 expression functionally alleviated the neuropathology of PD in wild-type but not in Sh2b1-deficient mice. This is the first study to examine the molecular underpinnings of SH2B1 against MPTP-induced neurodegeneration through cell autonomous promotion of neuronal survival in an in vivo PD model. Our findings reveal that SH2B1 antagonizes neurodegenerative pathology in PD via the SH2B1-HSC70-PLIN4 axis.


Mycotoxins in food systems in Sub Saharan Africa: A review.
Mycotoxins, toxic secondary metabolites of fungi are now recognised as major cause of food intoxications in Sub Saharan Africa (SSA). Aflatoxins, the most important of the group have been implicated in acute aflatoxicoses, carcinogenicity, growth retardation, neonatal jaundice and immunological suppression in SSA. The hot and humid tropical climate provides ideal condition for growth of toxigenicAspergillus spp, making food contamination to be widespread in SSA, with maize and groundnuts being the most contaminated. The available data suggests that cassava products (the most important African food) are not prone to aflatoxin contamination. Recent data on ochratoxin A produced by species ofAspergillus on grains have indicated the necessity for it to be monitored in SSA. Fumonisins represent the most importantFusarium mycotoxins in SSA, and surveillance data indicate very high contamination rates of almost 100% in maize samples from West Africa. Limited information exists on the occurrence of trichothecenes, while the data currently available suggest that zearalenone contamination seems not to be a problem in SSA. The strategies under investigation to mitigate the mycotoxin problem in SSA include education of the people on the danger of consuming mouldy foods, pre and post harvest management strategies with emphasis on biological control, use of plant products to arrest fungal growth during storage, enterosorbent clay technology, and the search for traditional techniques that could reduce/detoxify mycotoxins during food processing.


Obesity, overweight and ethnicity.
This article describes the prevalence of self-reported overweight and obesity, based on body mass index (BMI), by ethnicity and examines the influence of time since immigration within and between ethnic groups. Results are based on data from two cycles of Statistics Canada's Canadian Community Health Survey, conducted in 2000/01 and 2003. Weighted prevalences of overweight (BMI > or =25) and obesity (BMI > or =30) were calculated by sex and ethnicity for the population aged 20 to 64. Multiple logistic regression models were used to examine associations between overweight/obesity and ethnicity, and within and between ethnic groups based on time since immigration, controlling for age, household income, education and physical activity. Aboriginal men and women had the highest prevalences of overweight and obesity; East/Southeast Asians, the lowest. Independent of age, household income, education and physical activity, Aboriginal people had elevated odds of overweight and obesity, compared with Whites; South Asians and East/Southeast Asians had significantly lower odds. Recent immigrants (10 years or less) had significantly lower prevalences of overweight, compared with non-immigrants, but this difference tended to disappear over time.


Biofilm formation on restorative materials and resin composite cements.
Monolithic zirconia, polymer-infiltrated ceramic and acrylate polymer cemented with resin composite cement have recently been identified as prosthetic treatment options for zirconia implants. The aim of the present study is to determine in vitro, to what extent bacteria adhere to these materials. Disks made of zirconia (Vita YZ [YZ]), polymer-infiltrated ceramic (Vita Enamic [VE]), acrylate polymer (Vita CAD-Temp [CT]), self-adhesive cement (RelyX Unicem 2 Automix [RUN]) and of two different adhesive cements (RelyX Ulimate [RUL] and Vita Adiva F-Cem [VAF]) were produced. The biofilm formation of three bacterial species (Streptococcus sanguinis, Fusobacterium nucleatum, Porphyromonas gingivalis) on each material was assessed over 72h using a flow chamber system. The biofilms were quantified by crystal violet staining (optical density 595nm) and visualized using SEM. The inorganic composition of the different materials was analyzed and the wettability of the specimens was measured. For the restorative materials lowest biofilm formation was found on CT: OD 0.5±0.1, followed by VE: OD 0.8±0.1 and YZ: OD 1.4±0.3. The biofilm formation on resin composite cements was significantly lower on VAF: OD 0.6±0.1 than for RUL: OD 0.9±0.1 and RUN: OD 1.0±0.1. A high wettability of the specimens with saliva/serum mixture tended to result in a higher biofilm formation. Correlations were obtained between the organic/inorganic composition of the materials and the polar/dispersive part of the surface free energy. Three-species biofilm formation on restorative and cement materials strongly relies on the materials composition. If the restorative material CT and cement VAF also prevent excessive biofilm formation in a clinical situation should be further investigated.


Glutamatergic mechanisms for speed control and network operation in the rodent locomotor CpG.
Locomotion is a fundamental motor act that, to a large degree, is controlled by central pattern-generating (CPG) networks in the spinal cord. Glutamate is thought to be responsible for most of the excitatory input to and the excitatory activity within the locomotor CPG. However, previous studies in mammals have produced conflicting results regarding the necessity and role of the different ionotropic glutamate receptors (GluRs) in the CPG function. Here, we use electrophysiological and pharmacological techniques in the in vitro neonatal mouse lumbar spinal cord to investigate the role of a broad range of ionotropic GluRs in the control of locomotor speed and intrinsic locomotor network function. We show that non-NMDA (non-NMDARs) and NMDA receptor (NMDAR) systems may independently mediate locomotor-like activity and that these receptors set different speeds of locomotor-like activity through mechanisms acting at various network levels. AMPA and kainate receptors are necessary for generating the highest locomotor frequencies. For coordination, NMDARs are more important than non-NMDARs for conveying the rhythmic signal from the network to the motor neurons during long-lasting and steady locomotor activity. This study reveals that a diversity of ionotropic GluRs tunes the network to perform at different locomotor speeds and provides multiple levels for potential regulation and plasticity.


A meta-analysis examining the association between the MUC5B rs35705950 T/G polymorphism and susceptibility to idiopathic pulmonary fibrosis.
To explore whether the mucin (MUC) 5B rs35705950 T/G polymorphism confers susceptibility to idiopathic pulmonary fibrosis (IPF). A meta-analysis was conducted to determine associations between the MUC5B rs35705950 T/G polymorphism and either IPF or connective tissue disease-associated interstitial lung disease (CTD-ILD). We used the allele contrast, recessive, dominant, and additive models. A total of 12 IPF studies comprising 2859 patients and 6901 controls and four CTD-ILD studies involving 903 patients and 3306 controls were included in the meta-analysis. There was a significant association between the Tallele of the MUC5B rs35705950 polymorphism and IPF in all subjects (OR 3.768, 95 % CI 2.935-4.836, p < 1.0 × 10(-8)). Analysis after stratification by ethnicity indicated that the Tallele was significantly associated with IPF in Europeans and Asians (OR 3.728, 95 % CI 2.858-4.863, p < 1.0 × 10(-8); OR 4.334, 95 % CI 2.186-8.594, p = 2.6 × 10(-6)). However, there was no association between the Tallele and CTD-ILD in all subjects (OR 1.130, 95 % CI 0.937-1.363, p = 0.200), and in Europeans and Asians. Subgroup analysis by CTD type revealed no association between the Tallele and systemic sclerosis-associated ILD (SSc-ILD) and other CTD-ILDs. The MUC5B rs35705950 T/G polymorphism confers susceptibility to IPF in Europeans and Asians, but is not associated with susceptibility to CTD-ILD.


Goal-specific brain MRI harmonization.
There is significant interest in pooling magnetic resonance image (MRI) data from multiple datasets to enable mega-analysis. Harmonization is typically performed to reduce heterogeneity when pooling MRI data across datasets. Most MRI harmonization algorithms do not explicitly consider downstream application performance during harmonization. However, the choice of downstream application might influence what might be considered as study-specific confounds. Therefore, ignoring downstream applications during harmonization might potentially limit downstream performance. Here we propose a goal-specific harmonization framework that utilizes downstream application performance to regularize the harmonization procedure. Our framework can be integrated with a wide variety of harmonization models based on deep neural networks, such as the recently proposed conditional variational autoencoder (cVAE) harmonization model. Three datasets from three different continents with a total of 2787 participants and 10,085 anatomical T1 scans were used for evaluation. We found that cVAE removed more dataset differences than the widely used ComBat model, but at the expense of removing desirable biological information as measured by downstream prediction of mini mental state examination (MMSE) scores and clinical diagnoses. On the other hand, our goal-specific cVAE (gcVAE) was able to remove as much dataset differences as cVAE, while improving downstream cross-sectional prediction of MMSE scores and clinical diagnoses.


Enhanced lipidic algae biomass production using gas transfer from a fermentative Rhodosporidium toruloides culture to an autotrophic Chlorella protothecoides culture.
In order to produce single-cell oil for biodiesel, a yeast and a microalga were, for the first time, grown in two separate reactors connected by their gas-phases, taking advantage of their complementary nutritional metabolisms, i.e., respiration and photosynthesis. The yeast Rhodosporidium toruloides was used for lipid production, originating a carbon dioxide-enriched outlet gas stream which in turn was used to stimulate the autotrophic growth of Chlorella protothecoides in a vertical-alveolar-panel (VAP) photobioreactor. The microalgal biomass productivity was 0.015 gL(-1)h(-1), and its lipid productivity attained 2.2 mgL(-1)h(-1) when aerated with the outlet gas stream from the yeast fermenter. These values represent an increase of 94% and 87%, respectively, as compared to a control culture aerated with air. The CO2 bio-fixed by the microalgal biomass reached an estimated value of 29 mgL(-1)h(-1) in the VAP receiving the gas stream from the fermenter, a value 1.9 times higher than that measured in the control VAP.


Circular RNA circ_0001445 alleviates the ox-LDL-induced endothelial injury in human primary aortic endothelial cells through regulating ABCG1 via acting as a sponge of miR-208b-5p.
Coronary artery disease (CAD) originates from the blockage of the inner walls of the coronary arteries due to a plaque buildup. Circular RNA (circRNA) circ_0001445 has been reported to be downregulated in patients with a higher coronary atherosclerotic burden. This study is designed to explore the role and mechanism of circ_0001445 on the oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell damage. Circ_0001445, microRNA-208b-5p (miR-208b-5p), and ATP-binding cassette sub-family G member 1 (ABCG1) levels were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Inflammatory cytokines levels, cell viability, proliferation, migration were detected by Enzyme-linked immunosorbent assay (ELISA) kits, Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), and transwell assays, respectively. Protein levels were determined by western blot assay. The binding between miR-208b-5p and circ_0001445 or ABCG1 was predicted by circBank or TargetScan, and then verified by a dual-luciferase reporter, RNA Immunoprecipitation (RIP), and RNA pull-down assays. Circ_0001445 and ABCG1 were decreased, and miR-208b-5p was increased in CAD patients and ox-LDL-treated HAECs. Also, circ_0001445 overexpression could weaken ox-LDL-triggered HAEC injury by boosting proliferation, migration, and repressing inflammation and extracellular matrix (ECM). Mechanically, circ_0001445 directly targeted miR-208b-5p. Furthermore, miR-208b-5p mediated the modulation of circ_0001445 in ox-LDL-induced HAEC injury. ABCG1 acted as a direct target of miR-208b-5p, and the downregulation of miR-208b-5p relieved ox-LDL-induced HAEC damage by interacting with ABCG1. Additionally, circ_0001445 regulated ABCG1 expression by sponging miR-208b-5p. Circ_0001445 could abate ox-LDL-mediated HAEC damage by the miR-208b-5p/ABCG1 axis, providing a novel insight into the pathogenesis and treatment of CAD.


Spatial distribution of allergenic pollen through a large metropolitan area.
For nearly a decade, the majority of the world's population has been living in cities, including a considerable percentage of people suffering from pollen allergy. The increasing concentration of people in cities results in larger populations being exposed to allergenic pollen at the same time. There is almost no information about spatial distribution of pollen within cities as well as a lack of information about the possible impact to human health. To obtain this increasing need for pollen exposure studies on an intra-urban scale, a novelty screening network of 14 weekly changed pollen traps was established within a large metropolitan area-Berlin, Germany. Gravimetric pollen traps were placed at a uniform street-level height from March until October 2014. Three important allergenic pollen types for Central Europe-birch (Betula), grasses (Poaceae), and mugwort (Artemisia)-were monitored. Remarkable spatial and temporal variations of pollen sedimentation within the city and the influences by urban local sources are shown. The observed differences between the trap with the overall highest and the trap with the overall lowest amount of pollen sedimentation were in the case of birch pollen 245%, grass pollen 306%, and mugwort pollen 1962%. Differences of this magnitude can probably lead to different health impacts on allergy sufferers in one city. Therefore, pollen should be monitored preferably in two or more appropriate locations within large cities and as a part of natural air quality regulations.


The smallest worthwhile effect is superior to the MCID for estimating acceptable benefits of knee arthroplasty.
Traditionally, the minimal clinically important difference (MCID) is used to judge the meaningfulness of outcomes in total knee arthroplasty (TKA). However, MCID estimates do not consider patient costs, potential side effects, and inconveniences. MCIDs vary substantially across TKA studies and have several conceptual and psychometric problems. A more scientifically sound alternative for estimating benefits patients expect TKA is the smallest worthwhile effect (SWE), measured with the benefit-harm trade-off method. We recruited 121 participants and followed them for 6 months after surgery. All participants completed Knee Injury and Osteoarthritis Score (KOOS) Pain and Function, and underwent an interview using the benefit-harm trade-off method. The absolute SWE at the 50th percentile (approximating the average patient) was 31 points KOOS Pain improvement and 28 points for KOOS Function, daily living. Construct validity was supported with strong associations between meeting SWE (yes or no) and satisfaction with 6-month outcome (yes or no) using Pearson Chi Square (24.5, P < 0.001). Current MCID-based methods for interpreting KOOS change scores have flaws while the SWE method is conceptually superior. We determined the SWE of TKA that would justify the costs, risks, and inconveniences of surgery. SWE estimates suggest that patients expect a substantially greater change in KOOS scores than would be expected using MCID estimates. Clinicians can use SWE estimates when discussing likely outcomes and potential TKA benefits and risks with their patients.


Stereoselective synthesis of all stereoisomers of orthogonally protected cyclobutane-1,2-diamine and some chemoselective transformations.
The four stereoisomers of protected cyclobutane-1,2-diamine have been prepared in an enantio- and diastereocontrolled manner through stereodivergent synthetic routes starting from a half-ester as a common chiral precursor. Orthogonal protection allows the chemoselective manipulation of both amino groups as shown in this work.


Elastofibroma. A correlated light and electron microscopic study.
Four cases of elastofibroma located in the subscapular region of 3 men aged 66, 74, and 83 years, and a woman 70 years old are reported. A correlated light and electron microscopic study including ultrastructural examination of Verhoeff's iron-hematoxylin (VIH)-stained sections was performed. Light microscopically, the elastofibromas were characterized by connective tissue built up by collagen fibers and sclerotic masses mingled with numerous fibers and globules of elastin material. In one micron thick Epon sections these elastin fibers often revealed a central axis surrounded by a mantle composed of periodic segments giving them a necklace-like appearance. The ultrastructural findings of these elastin structures, stained with VIH, and the appearance of the stroma cells and their relation to the elastin indicate that elastofibroma is a non-neoplastic reactive lesion in which elastin material is synthesized by the stromal cells and predominantly laid down around preexisting elastic fibers.


Practice nurses' perceptions of their immunization role and strategies used to promote measles, mumps, and rubella vaccine uptake in 2014 - 2018: A qualitative study.
The aim of this study was to explore which aspects of their role practice nurses perceive to be most influential and the strategies they employ to promote the MMR vaccine. Qualitative study employing in depth interviews. Fifteen London based practice nurses, nine in 2014 and six in 2018, took part in semi-structured interviews that were audio recorded and transcribed verbatim. Qualitative content analysis was used to systematically manage, analyse, and identify themes. Analysis of data identified aspects of their role practice nurses perceived to be most influential (the themes) including: promoting vaccination, assisting parents' to make informed decisions, and provided insight into how they used specific strategies to achieve these in practice. These themes were consistent over both phases of the study. The findings provide an understanding of: (i) the practice nurses perceptions of the most important aspects of their role when promoting the measles, mumps, and rubella vaccine; and (ii) the strategies they implemented in practice to achieve these. The latter included assisting parents in their immunization decisions and was facilitated by practice nurses engaging with parents to provide relevant evidence to address parent queries, dispel misconceptions and tailor strategies to promote the measles, mumps, and rubella vaccine. This study addresses the paucity of literature available that specifically explores practice nurses' perceptions of their role concerning the measles, mumps, and rubella vaccine. The findings reveal how practice nurses promote the measles, mumps, and rubella vaccine by identifying strategies to enable parents to make informed decisions. At a time of an increasing incidence of measles, practice nurses have an important public health role in achieving herd immunity levels for measles, mumps, and rubella.


Effects of benzodiazepine agonist, inverse agonist and antagonist drugs in the mouse staircase test.
This study examined the effects of the benzodiazepine agonist midazolam and inverse agonist noreleagnine independently and in conjunction with the antagonist flumazenil in the mouse staircase test. According to this paradigm, the numbers of steps ascended (NSA) and rears (NR) reflect locomotor activity and anxiety, respectively. Midazolam reduced NR at doses that did not affect NSA; this NR-lowering effect was blocked by flumazenil. Noreleagnine increased NR at doses that did not affect NSA; this NR-elevating effect was also blocked by flumazenil. Effective antagonist doses of flumazenil alone had no effect on NR or NSA. The exactly opposite effects of midazolam and noreleagnine on NR and their antagonism by flumazenil are consistent with the postulated activities of these drugs at benzodiazepine receptors.


In-Hospital Amantadine Does Not Improve Outcomes After Severe Traumatic Brain Injury: An 11-Year Propensity-Matched Retrospective Analysis.
Disruption of dopamine neurotransmission is associated with functional impairment after severe traumatic brain injury (sTBI). This has prompted the study of dopamine agonists, such as amantadine, to assist recovery of consciousness. Randomized trials have mostly addressed the posthospital setting, with inconsistent findings. Therefore, we evaluated the efficacy of early amantadine administration on recovery of consciousness after sTBI. We searched the medical records of all patients with sTBI admitted to our hospital between 2010 and 2021 who survived 10 days postinjury. We identified all patients receiving amantadine and compared them with all patients not receiving amantadine and a propensity score-matched nonamantadine group. Primary outcome measures included discharge Glasgow Coma Scale, Glasgow Outcome Scale-Extended score, length of stay, mortality, recovery of command-following (CF), and days to CF. In our study population, 60 patients received amantadine and 344 did not. Compared with the propensity score-matched nonamantadine group, the amantadine group had no difference in mortality (86.67% vs. 88.33%, P = 0.783), rates of CF (73.33% vs. 76.67%, P = 0.673), or percentage of patients with severe (3-8) discharge Glasgow Coma Scale scores (11.11% vs. 12.28%, P = 0.434). In addition, the amantadine group was less likely to have a favorable recovery (discharge Glasgow Outcome Scale-Extended score 5-8) (14.53% vs. 16.67%, P < 0.001), had a longer length of stay (40.5 vs. 21.0 days, P < 0.001), and had a longer time to CF (11.5 vs. 6.0 days, P = 0.011). No difference in adverse events existed between groups. Our findings do not support the early administration of amantadine for sTBI. Larger inpatient randomized trials are necessary to further investigate amantadine treatment for sTBI.


Synthesis and characterization of SnO2/rGO nanocomposite for an efficient photocatalytic degradation of pharmaceutical pollutant: Kinetics, mechanism and recyclability.
The SnO2 and SnO2/rGO nanostructures were successfully synthesized using the facile hydrothermal synthesis technique. The prepared nanostructures were well studied using different techniques such as XRD, XPS, UV-DRS, FT-IR, EDX, SEM and HR-TEM analysis. The crystalline nature of SnO2 and SnO2/rGO was confirmed by the XRD technique. The formation of highly pure SnO2 and SnO2/rGO nanostructures was confirmed by EDX analysis. The morphological results show the good agglomeration of several spherical nanoparticles. The optical properties were studied through the UV-DRS technique and the bandgap energies of SnO2 and SnO2/rGO are estimated to be 3.12 eV and 2.71 eV, respectively. The photocatalytic degradation percentage in presence of SnO2 and SnO2/rGO against RhB was found to be 96% and 98%, respectively. The degradation of TTC molecules was estimated as 90% and 88% with SnO2/rGO and SnO2, respectively. The degradation of both RhB and TTC molecules was well suited with the pseudo-first-order kinetics. The results of successive experiments clearly show the enhancement in the photocatalytic properties in the SnO2/rGO nanostructures.


Molecular cloning and characterization of a novel UDP-GlcNAc:GalNAc-peptide beta1,3-N-acetylglucosaminyltransferase (beta 3Gn-T6), an enzyme synthesizing the core 3 structure of O-glycans.
The core 3 structure of the O-glycan, GlcNAcbeta1-3GalNAcalpha1-serine/threonine, an important precursor in the biosynthesis of mucin-type glycoproteins, is synthesized by UDP-N-acetylglucosamine:GalNAc-peptide beta1,3-N- acetylglucosaminyltransferase (beta3Gn-T; core 3 synthase). The core 3 structure is restricted in its occurrence to mucins from specific tissues such as the stomach, small intestine, and colon. A partial sequence encoding a novel member of the human beta3Gn-T family was found in one of the data bases. We cloned a complementary DNA of this gene and named it beta3Gn-T6. The putative amino acid sequence of beta3Gn-T6 retains the beta3Gn-T motifs and is predicted to comprise a typical type II membrane protein. The soluble form of beta3Gn-T6 expressed in insect cells showed beta3Gn-T activity toward GalNAcalpha-p-nitrophenyl and GalNAcalpha1-serine/threonine. The beta1,3-linkage between GlcNAc and GalNAc of the enzyme reaction product was confirmed by high performance liquid chromatography and NMR analyses. beta3Gn-T6 effectively transferred a GlcNAc to the GalNAc residue on MUC1 mucin, resulting in the synthesis of a core 3 structure. Real time PCR analysis revealed that the beta3Gn-T6 transcript was restricted in its distribution, mainly to the stomach, colon, and small intestine. We concluded that beta3Gn-T6 is the most logical candidate for the core 3 synthase, which plays an important role in the synthesis of mucin-type O-glycans in digestive organs.


Assessment of care of children with sickle cell disease: implications for neonatal screening programmes.
To assess the quality of care provided by hospitals for young children with sickle cell disease. Retrospective survey. Teaching hospital in London. 31 Children (mean age 4 years 4 months, range 4 months to 7 years 5 months) born with sickle cell disease between 1978 and 1985 identified from Hospital Activity Analysis data, an outpatient diagnostic register, and registers of the haematology department. Eight had been diagnosed on neonatal screening and at least four of these had not been followed up. Aspects of quality of outpatient care (blood testing, clinic attendance, and prophylactic drug treatment) and family care (adequate support and carers' knowledge about the disease) as assessed by reviewing the notes and administering a semistructured questionnaire to the carers, in relation to a devised list of standards deemed necessary to ensure achievement of the aims of screening. There were 93 outpatient attendances during the previous 12 months, but 13 children had not attended at least every six months and four not at all for more than a year. Only eight children had had three of the blood tests considered to be necessary for good care; three had had none. Prophylactic treatment with penicillin and folic acid was erratic; three children with sickle cell anaemia were not receiving regular prophylactic penicillin. Diagnosis of sickle cell disease on neonatal screening must be linked with follow up to ensure optimal management.


Spitz nevus and melanoma: evaluation with dermoscopy and reflectance confocal microscopy.
Spitz nevi (SN) include a wide range of benign melanocytic nevi, which are controversial due to their morphologic resemblance to melanoma (MM). To describe dermoscopic and reflectance confocal microscopic (RCM) features of SN compared to MM and assess the RCM utility in the differential diagnosis. We performed a multicentre retrospective analysis of MM and SN evaluated with dermoscopy and reflectance confocal microscopy. Three RCM mosaics were obtained for each lesion. Nine dermoscopic and twenty-one microscopic features were assessed for each lesion. A total of 26 lesions (15 SN and 11 MM) were included. Dermoscopically, most SN showed a "starburst" pattern. Asymmetry was marked in 8 MM. There were 6 dermoscopic features significantly more prevalent in MM than in SN. RCM showed that an atypical honeycomb pattern, atypical infiltration, and disarray of the epidermis were significant for MM. SN mostly revealed a typical honeycomb pattern. At the DEJ, most of SN had a meshwork pattern; MM revealed an atypical meshwork pattern. Atypical cells and sheet-like structures were observed in most MM. A combined approach using dermoscopy and RCM supports the differential diagnosis of SN and MM. Although our study showed some significant differences between SN and MM in RCM, further research on a larger group should be considered.


Electropolymerized tyrosine-based thin films: selective cell binding via peptide recognition to novel electropolymerized biomimetic tyrosine RGDY films.
We have created thin films by cyclic voltammetry (CV) electropolymerizations of the following phenolic functional group-based monomers: phenol; tyrosineamide; the tetrapeptide RGDY-containing the integrin membrane adhesion protein recognition tripeptide RGD; RDGY, a nonsense control tetrapeptide; and 1:3 mixtures of tyrosineamide with the two tetrapeptide monomers. The film formation process and description of the film properties were obtained by repetitive CV cycling using the oscillating quartz frequency shift, Deltaf, and motional resistance shift, DeltaR, parameters obtained with the electrochemical quartz crystal microbalance technique. Only the poly(phenol) film exhibited close chain packing-based self-limiting behavior, where all film synthesis ceased after approximately 7 CV cycles. All other films continued to form by electropolymerization with successive CV cycles out to the maximum cycle number (30 cycles) we measured. All of the films exhibited little energy dissipation behavior. Using the quartz crystal microbalance, we next compared the time course of cell attachment with the washed films and demonstrated that cells bound best to films in the following order: RGDY sense peptide:tyrosineamide films>RDGY nonsense peptide:tyrosineamide films=tyrosineamide films>phenol films. Cell enumeration after washing and trypsinization revealed firm protein-based cell attachment to the underlying extracellular matrix for the RGDY-containing films. These sense peptide films bound and retained two- to fivefold as many cells as the other films, with cells exhibiting a normal morphology. These results suggest the operation of specific cell attachment to the electropolymerized films via the RGD binding site for cellular integrin membrane proteins. The electropolymerization method we studied here forms a cassette system for creating electropolymerized films tailored to specific attachment of different cell types by varying the electropolymerized Y(tyrosine)-containing recognition peptide.


Plasma volume estimation in severely ill infants and children using a simplified Evans blue method.
Plasma volume was measured using Evans blue dye and 125iodinated human serum albumin (RIHSA) simultaneously in order to evaluate the accuracy of a simplified Evans blue method recommended by Nielsen and Nielsen (1962). 9 studies were performed in 8 newborn infants weighing 1.07 to 2.85 kg and 16 studies in 14 patients aged 6 months to 14 years suffering from severe circulatory disturbances. In 20 studies, plasma volumes measured by Evans blue and by RIHSA agreed within +/-5%, and in all the studies within +/-10%. The Evans blue method yielded higher plasma volumes than the RIHSA method in 19 instances. The mean difference (paired t-test) was only significant in newborn infants (+4.0+/-3.6%; P less than 0.05). The disappearance rates of Evans blue exceeded that of RIHSA in 16 studies, but this was only significant in the patients older than 6 months (+2.3+/-4.2%/h; P less than 0.05).


Effects of zinc on macromolecular synthesis and nuclear division during the yeast to mycelium transition in Sporothrix schenckii.
Zinc ions (10 mM) have been reported previously to inhibit the yeast to mycelium transition in Sporothrix schenckii. Yeast cells of this fungus were harvested, selected by filtration and allowed to form germ tubes in a basal medium with glucose in the presence of 10 mM zinc and the effects of this ion on protein, RNA and DNA synthesis and nuclear division recorded. All of these processes were affected by the addition of 10 mM zinc to the medium. Nevertheless, the inhibition of protein synthesis was observed earlier than that of RNA or DNA synthesis and was of a greater magnitude than that observed for both of these processes. Protein synthesis was inhibited within the first hour after inoculation, at which time this process begins in the control cells. RNA synthesis was inhibited during the 3 to 6 h interval after inoculation, that is, 3 h after the start of this process in the control cells. After 9 h of incubation, the inhibition of protein synthesis had reached its maximum at 70%, while that of RNA synthesis was only 52%. DNA synthesis was slightly inhibited, with maximum inhibition being observed 9 h after inoculation. Nuclear division in cells forming germ tubes in the presence of 10 mM zinc took place with a 3 h delay in relation to the control cells. These observations suggest that the inhibition of protein synthesis might be the most important mechanism by which zinc inhibits the yeast to mycelium transition in S. schenckii.


Nonheme ferric hydroperoxo intermediates are efficient oxidants of bromide oxidation.
This work presents the first combined experimental and computational study that gives evidence of the electrophilic reactivity of a nonheme iron(III)-hydroperoxo species. We show that in contrast to their heme counterparts the nonheme iron(III)-hydroperoxo complexes are catalytically much more active and even more so than nonheme iron(IV)-oxo species.


Evaluation of the efficacy of micro-Magnetic Resonance Imaging compared with light microscopy to investigate the anatomy of modern and ancient waterlogged wood.
Light microscopy is the conventional method used to investigate wood anatomy, identify the wood taxon, and assess its conservation state. It generally requires the mechanical cut of thin sections from a sample to obtain informative images. When dealing with wooden artworks or ancient remains (e.g., archaeological waterlogged wood), it is important to avoid sample destruction. In this work the efficacy of micro-magnetic resonance imaging (μ-MRI) to investigate the anatomy of waterlogged wood is assessed in comparison with light microscopy. Images along the three anatomical directions (transverse, tangential and radial) of six modern wood species and one archaeological specimen of waterlogged wood (from the Neolithic site "La Marmotta") were obtained both by μ-MRI and light microscopy. μ-MRI images were acquired virtually selecting 2D slices along the three wood anatomical directions. A 3D reconstruction was derived from 2D μ-MRI images. Conventional light microscopy histology was obtained by manually cutting thin sections. To the best of our knowledge, this is the first study in which high-resolution MR images and light microscopy images of the three anatomical directions of seven wood species are compared. The non-destructive μ-MRI approach allows to investigate the 2D and 3D topological organization of the whole waterlogged wood sample up to a resolution of 8 μm. Although the optical microscope attains higher image resolutions and remains superior in the observation of wood diagnostic characters, multi-parametric μ-MRI provides physiological investigation complementary to light microscopy, giving information concerning both a single section and the whole volume of the sample. The presented study may represent a starting point for further improvements of μ-MRI techniques applied to the non-destructive investigation of waterlogged wood samples, especially those of interest for cultural heritage.


The effect of blood-flow-restricted interval training on lactate and H+ dynamics during dynamic exercise in man.
To assess how blood-flow-restricted (BFR) interval-training affects the capacity of the leg muscles for pH regulation during dynamic exercise in physically trained men. Ten men (age: 25 ± 4y; V˙O2max : 50 ± 5 mL∙kg-1 ∙min-1 ) completed a 6-wk interval-cycling intervention (INT) with one leg under BFR (BFR-leg; ~180 mmHg) and the other without BFR (CON-leg). Before and after INT, thigh net H+ -release (lactate-dependent, lactate-independent and sum) and blood acid/base variables were measured during knee-extensor exercise at 25% (Ex25) and 90% (Ex90) of incremental peak power output. A muscle biopsy was collected before and after Ex90 to determine pH, lactate and density of H+ -transport/buffering systems. After INT, net H+ release (BFR-leg: 15 ± 2; CON-leg: 13 ± 3; mmol·min-1 ; Mean ± 95% CI), net lactate-independent H+ release (BFR-leg: 8 ± 1; CON-leg: 4 ± 1; mmol·min-1 ) and net lactate-dependent H+ release (BFR-leg: 9 ± 3; CON-leg: 10 ± 3; mmol·min-1 ) were similar between legs during Ex90 (P > .05), despite a ~142% lower muscle intracellular-to-interstitial lactate gradient in BFR-leg (-3 ± 4 vs 6 ± 6 mmol·L-1 ; P < .05). In recovery from Ex90, net lactate-dependent H+ efflux decreased in BFR-leg with INT (P < .05 vs CON-leg) owing to lowered muscle lactate production (~58% vs CON-leg, P < .05). Net H+ gradient was not different between legs (~19%, P > .05; BFR-leg: 48 ± 30; CON-leg: 44 ± 23; mmol·L-1 ). In BFR-leg, NHE1 density was higher than in CON-leg (~45%; P < .05) and correlated with total-net H+ -release (r = 0.71; P = .031) and lactate-independent H+ release (r = 0.74; P = .023) after INT, where arterial [ HCO3- ] and standard base excess in Ex25 were higher in BFR-leg than CON-leg. Compared to a training control, BFR-interval training increases the capacity for pH regulation during dynamic exercise mainly via enhancement of muscle lactate-dependent H+ -transport function and blood H+ -buffering capacity.


Impact of herbicide Ametryn on microbial communities in mixed liquor of a membrane bioreactor (MBR).
Ametryn, which is a second generation herbicide, was introduced to a lab-scale MBR at a concentration of 1mg/L and a 20-40% removal was observed at HRT ranging from 7.8 to 15.6h for an average influent Ametryn concentration of 0.8 mg/L. Components of EPS (protein and carbohydrates) increased in the bioreactor and the observed biomass production reduced after the addition of Ametryn. In a batch study, GAC was added to MBR mixed liquor and removal of Ametryn via biodegradation and adsorption were measured. Five common bacterial colony types (Gram negative and positive bacilli and Gram negative cocci) were identified and three of these were resistant to Ametryn up to 5mg/L. GAC was found to be a very effective Ametryn adsorption medium and in some occasions Ametryn may have acted as a nutrient source for bacteria.


Audiogenic seizures evoked in DBA/2 mice induce c-fos oncogene expression into subcortical auditory nuclei.
Recently the nuclear proto-oncogene c-fos has been shown to be rapidly and transiently expressed following seizures in many types of epilepsies. Until now, immunohistochemical as well as in situ hybridization studies have reported that the dentate gyrus of the hippocampus and most of the cortical areas were invariably heavily labeled. In order to see whether this distribution was reproduced or not in a model of epilepsy which has been proved to not involve these structures, a study was performed on genetically epilepsy-prone DBA/2 mice. Here we show that following audiogenic seizures, c-fos oncoprotein is not expressed in cortical and limbic structures but rather mapped the subcortical auditory nuclei.


Optimization of transfection of green fluorescent protein in pursuing mesenchymal stem cells in vivo.
Green Fluorescent Protein (GFP) has been used as a marker of gene expression and a single cell marker in living organisms in cell biology studies. The important areas that GFP is used are expression levels of different genes in different organisms by inserting GFP in these genes and as a marker in living cells. In this study, we tried to optimize transfection of mesenchymal stem cells, (MSCs) used for regeneration of damaged tissues in animals, by GFP containing plasmid vector by which MSCs can be followed in vivo. To this aim, phM-GFP plasmid vector carrying GFP gene and effectene transfection reagent were used. The data revealed that twice transfection of MSCs resulted in higher expression of GFP for longer times as compared to once transfected MSCs. On the other hand, leaving the chemical transfection agents in the medium induced apoptosis after a while. As a conclusion we suggest the transfection of MSCs twice with 48 hours interval and removal of transfection agents after 8 hours which removed toxic and apoptotic effects of the chemicals.


High levels of fluctuating asymmetry in populations of Apodemus flavicollis from the most contaminated areas in Chornobyl.
Random deviations from the perfect symmetry of normally bilaterally symmetrical characters for an individual with a given genotype occur during individual development due to the influence of multiple environmental factors. Fluctuating asymmetry (FA) is often used as a measure of developmental instability, and can be estimated as the variance of the distribution of differences between the left and right sides. We addressed the question of whether levels of FA were elevated in radioactively contaminated populations living around Chornobyl compared to those in reference populations of the yellow-necked mouse (Apodemus flavicollis). In addition, we studied amounts of directional asymmetry (DA) when one side is larger than the other on average. There was a significant difference among populations, including reference populations, in the amount of both FA and DA. A higher level of FA was documented for the contaminated populations in close proximity to the failed Chornobyl reactor for both the asymmetry of size and shape. The FAs of size and shape were highest in populations from the most contaminated locations in the Chornobyl exclusion zone. Although the directional asymmetry of shape was also highest in the contaminated populations, it was not significantly different from those in most of the reference populations. Populations from less contaminated areas inside the Chornobyl exclusion zone did not express FA values different from those of the reference populations outside the affected area. FA of skulls of A. flavicollis may indicate the degree to which the level of radioactive contamination affects the development of animals at Chornobyl. However, the mechanisms leading to these effects are not clear and probably vary from population to population. There were significant correlations between the overall right to left differences for the Procrustes aligned shape configurations, centroid sizes, and intramuscular (137)Cs. Detectable effects of radiation on developmental stability probably start to occur between 0.132 and 0.297 microGy/h.


Evidence that PICALM affects age at onset of Alzheimer's dementia in Down syndrome.
It is known that individuals with Down syndrome develop Alzheimer's disease with an early age at onset, although associated genetic risk factors have not been widely studied. We tested whether genes that increase the risk of late-onset Alzheimer's disease influence the age at onset in Down syndrome using genome-wide association data for age at onset of dementia in a small sample of individuals (N = 67) with Down syndrome. We tested for association with loci previously associated with Alzheimer's disease risk and, despite the small size of the study, we detected associations with age at onset of Alzheimer's disease in Down syndrome with PICALM (β = 3.31, p = 0.011) and the APOE loci (β = 3.58, p = 0.014). As dementia in people with Down syndrome is relatively understudied, we make all of these data publicly available to encourage further analyses of the problem of Alzheimer's disease in Down syndrome.


Advantages and challenges in nanomedicines for chronic liver diseases: A hepatologist's perspectives.
Chronic liver diseases (CLD) are responsible for significant morbidity and mortality worldwide. CLD patients are at a high risk of developing progressive liver fibrosis, cirrhosis, hepatocellular carcinoma (HCC), and subsequent liver failure. To date, there is no specific and effective therapies exist for patients with various forms of CLD. The application of nanotechnology has emerged as a rapidly developing area of interest for the safe and target-specific delivery of poorly aqueous soluble hepatoprotective agents and nucleic acids (siRNA/miRNAs) in CLD. The nanoparticle combination improves bioavailability and plasma stability of drugs with poor aqueous solubility. However, the extent of successful functional delivery of nanoparticles into hepatocytes is often surprisingly low. High Kupffer cells interaction reduces the nanomedicine efficacy. During fibrosis, the extracellular matrix accumulation in the perisinusoidal space restricts nanoparticle delivery to hepatocytes. The availability and uptake of nanoparticles exposure to different cells in the liver microenvironment is as Kupffer cells > sinusoidal endothelial cells > HSCs > hepatocytes. The most widely used strategy to reduce nanoparticles and macrophages interaction is to coat the particle surface with polyethylene glycol. The cationic charged nanoparticles have increased hepatocyte delivery by increased cellular interaction by disrupting the endosomal system via their pH buffering capacity. The immune clearance and toxicity of nanoparticles are mainly unpredictable. Therefore, more elaborate knowledge on exact cellular uptake and intracellular accumulation, trafficking, and endosomal sorting of nanoparticle is the need of the hour to improve the rational carrier design.


A subpopulation of olivocerebellar projection neurons express neuropeptide Y.
By neuropeptide Y (NPY) immunohistochemistry and in situ hybridization histochemistry with digoxigenin-labeled oligonucleotide probes for this peptide, a subpopulation of neurons in the caudal portions of the rat dorsal and medial accessory olives were found to express NPY immunoreactivity and mRNA. In the cerebellum, NPY-immunolabeled climbing fibers were distributed to the flocculus and parts of vermal cortex. The results suggest particular association of NPY with the climbing fiber system presumably mediating spinal, visual and vestibular inputs.


Investigating a possible link between antiseptic treatment and the increased occurrence of daptomycin-resistant Staphylococcus aureus.
Because of a steady increase in the detection of daptomycin-resistant (DAP-R) Staphylococcus aureus at three medical centres in Cologne, Germany, molecular surveillance was established from June 2016 to June 2018 to investigate the causes of the emergence and spread of respective isolates. Seventy-five S. aureus isolates, both DAP-R and DAP-susceptible, were collected from 42 patients for further analysis. Broth microdilution was used to determine the MICs for DAP and polyhexamethylene biguanide/polyhexanide (PHMB). To investigate the effect of PHMB on the development of DAP resistance, we performed selection experiments with PHMB. All isolates studied were subjected to whole-genome sequencing. Epidemiological, clinical, microbiological and molecular data were analysed comparatively. Acquisition of DAP resistance was mainly observed in patients with acute and chronic wounds (40/42, 96.2%) treated with antiseptic (32/42, 76.2%) rather than systemic antibiotic therapy using DAP or vancomycin (7/42, 16.7%). DAP-R S. aureus had a diverse genetic background; however, within individual patients, isolates were closely related. At least three potential transmission events were detected. Most DAP-R isolates had concomitant elevated MICs for PHMB (50/54, 92.6%), and in vitro selection experiments confirmed that PHMB treatment is capable of generating DAP resistance. DAP resistance could be linked to 12 different polymorphisms in the mprF gene in the majority of clinical isolates (52/54, 96.3%) as well as in all in vitro selected strains. DAP resistance in S. aureus can occur independently of prior antibiotic therapy and can be selected by PHMB. Therefore, wound treatment with PHMB may trigger individual resistance development associated with gain-of-function mutations in the mprF gene.


The effects of soil properties on the turbidity of catchment soils from the Yongdam dam basin in Korea.
Environmental concerns have been raised that suspended solids in turbid water adversely affect human health, and that their removal increases in the cost of water treatment. The Yongdam dam reservoir, located in the southwestern region of Korea, is severely affected by inflowing turbid water after storms. In this study, soil samples were collected from 37 sites in the Yongdam upstream basin to investigate mineralogical and environmental factors associated with the turbidity potential of soils in water environments. Turbidity potential was estimated by measuring the turbidity of soil-suspension solutions after settling for 24 h. The mineralogy of the soils was dominated by four minerals-quartz, microcline, albite, and muscovite-with lesser amounts of hornblende, chlorite, kaolinite, illite, and mixed layer illite. The quartz content was the most variable of the soil mineralogy among the collected samples. Principal-components analysis (PCA) was used to examine relationships between turbidity potential and other soil properties. The variables considered in the PCA included turbidity potential, quartz content, albite content, mean size of soil particles, clay content, clay mineral content, zeta potential, conductivity, and pH of the soil-suspension solution. The first two components of the PCA explained 52% of the overall variation of the selected variables. The first component was possibly explained by physical properties such as the size of the soil particles; the second was correlated with chemical properties of the soils, for example dissolution and extent of weathering. Closer examination of the PCA results revealed that the quartz content of the soils was negatively correlated with their turbidity potential. A linear correlation (r = 0.63) was obtained between measured turbidity potential and that predicted using multiple regression analysis based on the content of clay-sized particles, clay minerals, and quartz, and the conductivity of the soil-suspension solution.


Fructooligosaccharides and wheat bran fed at similar fermentation levels differentially affect the expression of genes involved in transport, signaling, apoptosis, cell proliferation, and oncogenesis in the colon epithelia of healthy Fischer 344 rats.
The influence of the source of fermentable material (FM) on the luminal concentrations of their end products and its effects on colon cell metabolism and disease susceptibility is not well characterized. We hypothesized that total fermentation but not the source (type) of FM would be the main factor in determining cellular /molecular outcomes in the healthy colon epithelia. The main aim of this study was to elucidate the role of two different sources of FM, fructooligosaccharides (FOS) and wheat bran (WB), on the expression of genes involved in short chain fatty acid (SCFA) transport, G-protein signaling, apoptosis, cell proliferation and oncogenesis in colon epithelia of healthy rats. Male Fischer 344 rats (n = 10/group) were fed AIN-93G control (0% FM) or experimental diets containing WB (~1%, 2%, or 5% FM) or FOS (~2%, 5%, or 8% FM). Rats were killed after 6 weeks and the colon mucosa was assessed for the expression of target genes using real-time quantitative polymerase chain reaction. By comparison to the control, dose-related changes of mRNA levels were found in rats fed FOS-based diets, including: (a) upregulation of three SCFA transporters (Smct2, Mct1 and Mct4) but downregulation of Mct2, (b) upregulation of Gpr109a and downregulation of Gpr120, Gpr43, Gprc5a, Rgs2 and Rgs16, (c) upregulation of apoptosis-related genes including Bcl2, Bcl2-like 1, Bak1, Caspase 3, Caspase 8 and Caspase 9, (d) downregulation of the oncogenes and metastasis genes Ros1, Fos, Cd44, Fn1 and Plau, and (e) downregulation of several genes involved in cellular proliferation including Hbegf, Hoxb13, Cgref1, Wfdc1, Tgm3, Fgf7, Nov and Lumican. In contrast, rats fed WB-based diets resulted in dose-related upregulation of mRNA levels of Smct2, Rgs16, Gprc5a, Gpr109a, Bcl2-like 1, Caspase 8, and Fos. Additionally, different gene expression responses were observed in rats fed FOS and WB at 2% and 5% FM. Over all, these gene changes elicited by FOS and WB were independent of the expression of the tumor suppressor Tp53. These results suggest that fermentation alone is not the sole determinant of gene responses in the healthy rat colon.


An eosinophil peroxidase activity assay accurately predicts eosinophilic chronic rhinosinusitis.
A definitive diagnosis of eosinophilic chronic rhinosinusitis (eCRS) requires invasive surgical tissue sampling and histologic enumeration of intact eosinophils. Eosinophil peroxidase (EPX) is an accurate biomarker of sinonasal tissue eosinophilia in CRS regardless of polyp status. A less invasive and rapid method that accurately identifies tissue eosinophilia would be of great benefit to patients. We sought to evaluate a new clinical tool that uses a nasal swab and colorimetric EPX activity assay to predict a diagnosis of eCRS. A prospective, observational cohort study was conducted using nasal swabs and sinonasal tissue biopsies obtained from patients with CRS electing endoscopic sinus surgery. Patients were classified as non-eCRS (n = 19) and eCRS (n = 35) on the basis of pathologically determined eosinophil counts of less than 10 or greater than or equal to 10 eosinophils/HPF, respectively. Swab-deposited EPX activity was measured and compared with tissue eosinophil counts, EPX levels, and CRS-specific disease metrics. EPX activity was significantly increased in patients with eCRS than in patients without eCRS (P < .0001). With a relative absorbance unit cutoff value of greater than or equal to 0.80, the assay demonstrated high sensitivity (85.7%) and moderate specificity (79.0%) for confirming eCRS. Spearman correlations between EPX activity and tissue eosinophil counts (rs = 0.424), EPX levels (rs = 0.503), and Lund-Kennedy endoscopy scores (rs = 0.440) in eCRS were significant (P < .05). This investigation evaluates a nasal swab sampling method and EPX activity assay that accurately confirms eCRS. This method could potentially address the unmet need to identify sinonasal tissue eosinophilia at the point-of-care, as well as to longitudinally monitor eosinophil activity and treatment response.


Cerebral cation shifts in hypoxic-ischemic brain damage are prevented by the sodium channel blocker tetrodotoxin.
We investigated the effect of the sodium channel blocker, tetrodotoxin, in two animal models of brain pathology. In the first, an acute model, we recorded the interstitial brain potential in the striatum of rats after cardiac arrest. The time of deflection of this potential, an indication of changes in cerebral cation concentrations, was determined in control rats, and in rats pretreated with intrastriatal tetrodotoxin. In control rats a deflection of the brain potential was noted 2 min after cardiac arrest; tetrodotoxin pretreatment delayed this deflection to about 5 min. The second, a survival model, was based on the Levine preparation in rats. A combination of ischemia and hypoxia produced unilateral, cerebral infarcts, which were characterized by a decrease of brain [K+], and by increases of [Ca2+] and [Na+] and thus of the Na+:K+ ratio. Data on the cation shifts, determined by chemical assay methods, were complemented by those of more conventional methods of assessment of brain damage, such as the determination of survival, of Evans blue staining, and of brain water content. Cation shifts could be prevented locally by tetrodotoxin. In conclusion, the drug can, at least partially, prevent the detrimental effects of an ischemic insult. In addition, our results showed that protective effects observed in the acute model may sometimes offer an indication of the effects to be expected in the survival model. Furthermore, the effect of tetrodotoxin on the brain potentials in the acute model showed that its protective action in the survival model may be brought about by delaying cell depolarization and by shortening the actual duration of the depolarized state. We conclude that Na+ influx and, consequently, neurotransmission may play a crucial role in the development of cerebral damage.


The possible existence of a heat-stable alkaline proteinase (HAP) in rat skeletal muscle.
1. A unique caseinolytic activity was found in the crude extract from chicken and rat skeletal muscle. Hardly any activity was detected at physiological assay temperatures at pH 8.0 but did well at around 60 degrees C. 2. The activity partially purified from rat skeletal muscle showed optimum pH at around 8.0 at 60 degrees C. It hardly hydrolyzed casein below 50 degrees C, but in the presence of 5 M urea it showed relatively high activity at 30 degrees C. The activity was completely stable at 50 degrees C for 1 hr. 3. The activity seems to be contained in a high mol. wt (450,000) protein from the elution volume and is due to cysteine proteinase from the effect of inhibitors. 4. The above properties agreed with those of the heat-stable alkaline proteinase (HAP) of fish purified homogeneously by electrophoresis. This seems to suggest that HAP may also exist in rat skeletal muscle.


Influence of midbrain stimulation on the excitability of neurons in the medial hypothalamus of the rat.
Extracellular action potentials were recorded from 1098 neurons in the medial hypothalamus of pentobarbital anesthetized male rats. Their excitability was analyzed after single 1 Hz stimulation of the midbrain periaqueductal gray (PAG) or adjacent reticular formation. Cells were also examined for their response to median eminence (ME), amygdala, lateral septum (LS) or anterior hypothalamic/preoptic area (AHA/POA) stimulation. Antidromic invasion from midbrain stimulation was recorded from 110 neurons. Eight of these neurons showed features of axon branching and displayed antidromic invasion from both midbrain and amygdala (2 cells) or AHA/POA (6 cells). Many neurons with midbrain projections displayed orthodromic responses to stimulation in the amygdala, but few responded to AHA/POA or LS stimulation. Midbrain stimulation evoked orthodromic responses from 99 medial hypothalamic neurons. Many of these cells also displayed orthodromic responses to amygdala or AHA/POA stimulation, whereas a small number were activated antidromically by stimulation in these sites. None of 42 neurons activated antidromically from median eminence stimulation were responsive to midbrain stimulation. These results provide electrophysiological evidence of reciprocal connections between medial hypothalamic and medial midbrain areas, and indicate that medial hypothalamic neurons with midbrain connections are subject to influences from other extrahypothalamic areas.


Efficient production of recombinant proteins in suspension CHO cells culture using the Tol2 transposon system coupled with cycloheximide resistance selection.
DNA recombination techniques in mammalian cells has been applied to the production of therapeutic proteins for several decades. To be used for commercial production, established cell lines should stably express target proteins with high productivity and acceptable quality for human use. In the conventional transfection method, the screening process is laborious and time-consuming since superior cell lines had to be selected from an enormous number of transfected cell pools and clonal cell lines with a wide variety of transgene insertion locations. In this study, we demonstrated that the combination of a Tol2 transposon system and cell selection by cycloheximide resistance is an efficient method to express therapeutic proteins, such as human antibody in suspension culture of Chinese hamster ovary cells. The resulting stable cell lines showed constant productivity and cell growth over a long enough cultivation periods for recombinant protein production. We anticipate that this approach will prove widely applicable to protein production in research and development of pharmaceutical products.


Efficacy of endoscopic ultrasound-guided tissue acquisition for solid pancreatic lesions 20 mm or less in diameter suspected as neuroendocrine tumors or requiring differentiation.
For non-functioning pancreatic neuroendocrine tumors (pNETs) ≤ 20 mm, most guidelines consider follow-up observations as an option; however, the various treatment strategies are defined by size alone, even though the Ki-67 index is important for malignancy grading. Endoscopic ultrasound-guided tissue acquisition (EUS-TA) is the standard for the histopathological diagnosis of solid pancreatic lesions; however, recent results for small lesions remain unclear. Therefore, we examined the efficacy of EUS-TA for solid pancreatic lesions ≤ 20 mm suspected as pNETs or requiring differentiation and the non-increase rate in tumor size in follow-up cases. We retrospectively analyzed data of 111 patients (median age = 58 years) with lesions ≤ 20 mm suspected as pNETs or requiring differentiation who underwent EUS-TA. All patients underwent specimen evaluation by rapid onsite evaluation (ROSE). EUS-TA led to a diagnosis of pNETs in 77 patients (69.4%) and tumors other than pNETs in 22 patients (19.8%). The histopathological diagnostic accuracy of EUS-TA was 89.2% (99/111) overall, 94.3% (50/53) for 10-20 mm lesions, and 84.5% (49/58) for ≤ 10 mm lesions, with no significant difference in diagnostic accuracy (p = 0.13). The Ki-67 index was measurable in all patients with a histopathological diagnosis of pNETs. Among 49 patients with a diagnosis of pNETs who were followed up, one patient (2.0%) showed tumor enlargement. EUS-TA for solid pancreatic lesions ≤ 20 mm suspected as pNETs or requiring differentiation is safe and has adequate histopathological diagnostic accuracy, suggesting that follow-up observations of pNETs with a histological pathologic diagnosis are acceptable in the short term.


A search for piroplasmids and spirochetes in threatened pudu (Pudu puda) and associated ticks from Southern Chile unveils a novel Babesia sp. and a variant of Borrelia chilensis.
Cervids are important hosts for ticks and although they are refractory to some tick-borne agents such as Borrelia, they do act as reservoirs for others such as Babesia. Babesia and Borrelia are commonly transmitted by Ixodes spp. associated with deer, and most of the knowledge on their biological cycles comes from northern latitudes of the globe. In this study, we performed genetic screenings to detect tick-borne agents in blood and Ixodes stilesi ticks collected from an insular population of threatened pudu (Pudu puda), a pygmy deer species that inhabits temperate rainforests of southern South America. Inferred by phylogenetic analyses for 18S rRNA, COI and cytb genes, our results unveiled a novel genospecies of Babesia (Babesia sp. pudui) genetically related to Babesia odocoilei, a species that infects Odocoileus virginianus deer in North America. Although blood of the deer was negative for Borrelia infection, multilocus sequencing typing performed in one I. stilesi tick revealed the occurrence of a novel genetic variant of Borrelia chilensis, differing 0.93% and 0.18% in flaB and pepX genes with the type of strain for the species, respectively. Such a genetic divergence could be the result of thousands of years of isolation because of recent glaciation events that separated pudus and their tick populations at Chiloé Island approximately 437,000 years ago. The finding of a Babesia sp. has no precedents for wild and domestic ungulates in Chile and shows a novel piroplasmid that must be considered now on in rehabilitation centres and zoos that attend pudu deer. Further research is now necessary to confirm pathogenic roles.


Comparison of non-invasive, scalp-recorded auditory steady-state responses in humans, rhesus monkeys, and common marmosets.
Auditory steady-state responses (ASSRs) are basic neural responses used to probe the ability of auditory circuits to produce synchronous activity to repetitive external stimulation. Reduced ASSR has been observed in patients with schizophrenia, especially at 40 Hz. Although ASSR is a translatable biomarker with a potential both in animal models and patients with schizophrenia, little is known about the features of ASSR in monkeys. Herein, we recorded the ASSR from humans, rhesus monkeys, and marmosets using the same method to directly compare the characteristics of ASSRs among the species. We used auditory trains on a wide range of frequencies to investigate the suitable frequency for ASSRs induction, because monkeys usually use stimulus frequency ranges different from humans for vocalization. We found that monkeys and marmosets also show auditory event-related potentials and phase-locking activity in gamma-frequency trains, although the optimal frequency with the best synchronization differed among these species. These results suggest that the ASSR could be a useful translational, cross-species biomarker to examine the generation of gamma-band synchronization in nonhuman primate models of schizophrenia.


Prepandemic cross-reactive humoral immunity to SARS-CoV-2 in Africa: Systematic review and meta-analysis.
To assess the evidence on the presence of antibodies cross-reactive with SARS-CoV-2 antigens in prepandemic samples from African populations. We performed a systematic review and meta-analysis of studies evaluating prepandemic African samples using pre-set assay-specific thresholds for SARS-CoV-2 seropositivity. In total, 26 articles with 156 datasets were eligible, including 3437 positives among 29,923 measurements (11.5%) with large between-dataset heterogeneity. Positivity was similar for anti-nucleocapsid (14%) and anti-spike antibodies (11%), higher for anti-spike1 (23%), and lower for anti-receptor-binding domain antibodies (7%). Positivity was similar, on average, for immunoglobulin M and immunoglobulin G. Positivity was seen prominently in countries where malaria transmission occurs throughout and in datasets enriched in malaria cases (14%, 95% confidence interval, 12-15% vs 2%, 95% confidence interval 1-2% in other datasets). Substantial SARS-CoV-2 reactivity was seen in high malaria burden with or without high dengue burden (14% and 12%, respectively), and not without high malaria burden (2% and 0%, respectively). Lower SARS-CoV-2 cross-reactivity was seen in settings of high HIV seroprevalence. More sparse individual-level data showed associations of higher SARS-CoV-2 cross-reactivity with Plasmodium parasitemia and lower SARS-CoV-2 cross-reactivity with HIV seropositivity. Prepandemic samples from Africa show high levels of anti-SARS-CoV-2 seropositivity. At the country level, cross-reactivity tracks especially with malaria prevalence.


Development of a combined storage medium for 7-day storage of platelet concentrates and 42-day storage of red cell concentrates.
Studies were carried out to examine whether a single additive solution could support both platelet and red cell storage. An ionically balanced electrolyte solution fortified with citrate, glucose and bicarbonate was used. This solution has previously been shown to provide good platelet viability with storage for up to 7 d. Optimization studies demonstrated that with adenine added to this solution (CSM), it also allowed for satisfactory preservation of in vitro red cell parameters for up to 49 d of storage. Confirmatory paired in vivo post-transfusion studies were carried out in which platelets and red cells obtained from the same donor were processed and stored in CSM on one occasion, and in CPD-plasma (platelets) and AS-1 (red cells) on another occasion. Five paired studies were conducted with platelets stored for 5 d and red cells for 42 d; another five paired studies with platelets stored for 7 d and red cells for 49 d. Except for a slight decrease in platelet survival (P less than 0.05) with storage in CSM, there were no statistically significant differences in post-transfusion recoveries and survivals between test and control media, demonstrating that both platelets and red cells may be satisfactorily stored in a combined single additive solution.


Prognostic Value of KRAS Mutation Subtypes and PD-L1 Expression in Patients With Lung Adenocarcinoma.
The prognostic value of different KRAS (Kirsten rat sarcoma viral oncogene) mutation subtypes and their association with programmed death ligand 1 (PD-L1) expression in lung adenocarcinoma (LADC) remain unclear. We examined the association of KRAS mutation subtypes with clinical outcomes and PD-L1 expression status. Patients diagnosed with KRAS-mutated LADC were evaluated for PD-L1 expression, cancer staging, overall survival (OS), and relapse-free survival. A cohort of 254 KRAS-mutated LADC patients (median follow-up, 17 months) was studied. The 3 major subtypes of KRAS mutations were G12C (46.1%), G12V (21.7%), and G12D (15.7%). We found that all these subtypes had no impact on cancer stages, brain metastasis at diagnosis, OS, and relapse-free survival. Among this cohort, 33% of 94 patients who had PD-L1 staining data available had PD-L1-positive disease (≥ 1% of tumor cells). PD-L1 expression status was not significantly different among the 3 major mutation subtypes. Of interest, among patients with G12C mutation, positive PD-L1 expression was associated with significantly shorter OS (median survival, 5.7 vs. 12.8 months, P = .007). In multivariable analysis, PD-L1 positivity remained as an adverse factor for OS, with hazard ratio of 4.44 (P = .0007). PD-L1 status did not affect OS in other subtypes of mutations. KRAS mutation subtype is not associated with patient clinical outcomes or PD-L1 expression status. However, PD-L1 positivity appears to negatively affect OS in LADC patients with G12C mutation. Further study is needed to confirm our observation and to determine if programmed cell death 1/PD-L1 antagonist may affect the clinical outcome of patients with different KRAS mutation subtypes.


Hodgkin's disease.
The epidemiological features of Hodgkin's disease (HD) suggest that it is a heterogeneous condition which may have different aetiologies in different age groups. The risk factors for the development of HD in young adults suggest that delayed exposure to a common infectious agent may be involved in this age group. Seroepidemiological studies have shown that HD patients have elevated antibody titres to Epstein-Barr virus (EBV) and the elevated titres have been shown to precede the diagnosis of HD. Recent molecular studies provide support for the idea that EBV is involved in the pathogenesis of HD. EBV genomes are consistently found in a proportion of tumour biopsies, the EBV-infected cells are clonal and the EBV genomes have been localized to Reed-Sternberg cells. Furthermore, EBV latent gene products are expressed by the Reed-Sternberg cells. The majority of HD samples from patients aged greater than 50 years and less than 15 years are EBV positive, whereas the minority (less than 15%) of samples from young adults contain detectable EBV DNA. The results suggest that EBV plays a role in HD in children and older adults but that other agents, possibly other viruses, are involved in young adults.