Isolation and characterization of a T-superfamily conotoxin from Conus litteratus with targeting tetrodotoxin-sensitive sodium channels.
A T-1-conotoxin, lt5d, was purified and characterized from the venom of vermivorous hunting cone snails Conus litteratus. The complete amino acid sequence of lt5d (DCCPAKLLCCNP) has been determined by Edman degradation. With two disulfide bonds, the calculated average mass is 1274.57 Da, which is confirmed by MALDI-TOF mass spectrometry (average mass 1274.8778). Under whole cell patch-clamp mode, lt5d inhibits tetrodotoxin-sensitive sodium currents on adult rat dorsal root ganglion neurons, but has no effects on tetrodotoxin-resistant sodium currents. The inhibition of TTX-sensitive sodium currents by lt5d was found to be concentration-dependent with the IC(50) value of 156.16 nM. Thus, this is the first T-superfamily conotoxin identified to block TTX-sensitive sodium channels.


Mechanisms of hypercholesterolaemia in glycogen storage disease type I: defective metabolism of low density lipoprotein in cultured skin fibroblasts.
Hyperlipidaemia is a feature of glycogen storage disease type I (GSD-I) (Levy et al.). High levels of LDL cholesterol (200 +/- 25 mg dl-1) and apo B (387 +/- 44 mg dl-1) were found in association with hypercholesterolaemia in GSD-I. Related causative factors might be attributed to overproduction and/or delayed removal of LDL. In this study, a possible alteration in the clearance of LDL was examined. Using cultured fibroblasts for LDL receptor activity, the following observations were made: 1. GSD-I fibroblasts revealed only a slight decrease in LDL binding (65 +/- 7) when compared with controls (74 +/- 4 ng mg-1 protein), however, LDL internalization (382 +/- 24 vs. 570 +/- 52 ng mg-1 protein) and proteolytic degradation (2082 +/- 280 vs. 2916 +/- 12.5 ng mg-1 protein) were significantly affected (P less than 0.01). 2. Binding, internalization and proteolytic degradation of LDL from GSD-I were compared with that of controls, and were found to be significantly lower (P less than 0.01). 3. Substitution of control lipoprotein-deficient serum (LPDS) by GSD-I LPDS further diminished the above processes (P less than 0.05). Our results demonstrate that increased plasma cholesterol in GSD-I is due to a decreased catabolism of LDL. The data suggest that the problem may well be multifactorial, due to diminished receptor expression, abnormal LDL composition and impaired LDL receptor interaction due to a circulating inhibitory factor.


Defect-Engineering-Enabled High-Efficiency All-Inorganic Perovskite Solar Cells.
The emergence of cesium lead iodide (CsPbI3 ) perovskite solar cells (PSCs) has generated enormous interest in the photovoltaic research community. However, in general they exhibit low power conversion efficiencies (PCEs) because of the existence of defects. A new all-inorganic perovskite material, CsPbI3 :Br:InI3 , is prepared by defect engineering of CsPbI3 . This new perovskite retains the same bandgap as CsPbI3 , while the intrinsic defect concentration is largely suppressed. Moreover, it can be prepared in an extremely high humidity atmosphere and thus a glovebox is not required. By completely eliminating the labile and expensive components in traditional PSCs, the all-inorganic PSCs based on CsPbI3 :Br:InI3 and carbon electrode exhibit PCE and open-circuit voltage as high as 12.04% and 1.20 V, respectively. More importantly, they demonstrate excellent stability in air for more than two months, while those based on CsPbI3 can survive only a few days in air. The progress reported represents a major leap for all-inorganic PSCs and paves the way for their further exploration in order to achieve higher performance.


Venous return from distal regions affects heat loss from the arms and legs during exercise-induced thermal loads.
To study the role of venous return from distal parts of the extremities in influencing heat loss from the more proximal parts, changes in mean skin temperature (Tsk) of the non-exercising extremities were measured by color thermography during leg and arm exercise in eight healthy subjects. Thirty minutes of either leg or arm exercise at an ambient temperature (Ta) of 20 degrees C or 30 degrees C produced a greatly increased blood flow in the hand or foot and a great increase in venous return through the superficial skin veins of the extremities. During the first 10 min of recovery from the exercise, blood flow to and venous return from the hand or foot on the tested side was occluded with a wrist or ankle cuff at a pressure of 33.3 kPa (250 mm Hg), while blood flow to the control hand or foot remained undisturbed. During the 10-min wrist occlusion, Tsk increased significantly from 28.3 degrees +/- 0.41 degrees C to 30.1 degrees +/- 0.29 degrees C in the control forearm, but remained at nearly the same level (28.0 degrees +/- 0.34 degrees C to 28.2 degrees +/- 0.25 degrees C) in the occluded forearm. In the legs, although Tsk on both sides was virtually identical (32.0 degrees +/- 0.31 degrees C, control vs 32.0 degrees +/- 0.36 degrees C, tested) before occlusion, Tsk on the control side (32.6 degrees +/- 0.27 degrees C) was significantly higher than that on the tested side (32.2 degrees +/- 0.21 degrees C) after ankle occlusion.(ABSTRACT TRUNCATED AT 250 WORDS)


Nanoformulated Bioactive Compounds Derived from Different Natural Products Combat Pancreatic Cancer Cell Proliferation [Retraction].
[This retracts the article DOI: 10.2147/IJN.S238256.].


Underuse of Clinical Decision Rules and d-Dimer in Suspected Pulmonary Embolism: A Nationwide Survey of the Veterans Administration Healthcare System.
The diagnosis of pulmonary embolism (PE) remains a challenge. CT pulmonary angiography (CTPA) for suspected PE has become the primary imaging modality, but concerns regarding overutilization, overdiagnosis, radiation, and costs have led to algorithms that combine a clinical decision rule (CDR) and highly sensitive d-dimer to identify patients in whom PE can be safely excluded without further studies. This has been identified as a top five Choosing Wisely recommendation in pulmonary medicine, but adherence is modest at best and actual utilization is unknown. Therefore, a survey was conducted to determine the prevalence of this approach in the Veterans Administration (VA) healthcare system. A web-based questionnaire survey (SurveyGizmo.com) was developed and validated to query the utilization of CDR ± d-dimer in suspected PE. Key stakeholders identified from national VA mailing lists of radiology, pulmonary, and emergency medicine chiefs were sent an email describing the survey and provided a link for response. This study was reviewed and approved by our local institutional review board and accessing the link represented consent for participation. No personally identifiable data were collected and a drawing for a gift card was provided as an incentive. There were a total of 159 responses, with 120 fully completed surveys for analysis. The majority of respondents were chiefs (63%) with 11+ years of experience (80%), from hospitals with house staff (86%) and an emergency department (97%). Respondents were from emergency medicine (31%), pulmonary (27%), radiology (26%), and other departments (9%). The overwhelming majority of respondents (85%) did not require results of a CDR ± d-dimer before ordering a CTPA. Only 6.7% required a CDR + d-dimer, with others requiring either only a CDR (5.8%) or d-dimer (2.5%). The most common CDR was the Wells score, with only one using the Pulmonary Embolism Rule-Out Criteria. Nine of 18 (50%) regional Veterans Integrated Service Networks reported at least one site requiring a CDR before CTPA. An average of 9.6 CTPAs were estimated to be performed per week. Sorted by CDR and d-dimer use, 8 (CDR + d-dimer), 6.9 (CDR only), 8 (d-dimer only), 10.1 (no requirements) CTPA studies were performed weekly. The average CTPA yield for PE was estimated at 11.9% (CDR + d-dimer), 8% (CDR only), 2.5% (d-dimer only), and 7.6% (no requirements). The vast majority of hospitals within the VA system do not use a CDR ± d-dimer in the evaluation of patients with suspected PE. Utilization of a CDR and d-dimer may decrease CTPA utilization and increase yield, but this assessment is limited by the scope of the survey. CDR-guided strategies are recommended in the evaluation of suspected PE. Adherence within the VA healthcare system is very low. Further investigation is warranted to better characterize and improve the adherence to CDR-guided strategies and CTPA utilization.


Utilization and value addition of copper tailing as an extender for development of paints.
The present study deals with characterization of copper tailing waste, test for possibilities of hazards and its potential reuse as an extender in paints. The waste is a siliceous material containing aluminum oxide, iron oxide and sulphate in significant concentrations. In the primary stage waste is acidic in nature, which makes it unsuitable for paint. This acidity is removed from the waste by simple sieving and grinding. The prepared mass was characterized for basic properties of an extender like oil absorption, specific gravity, pH, etc. Toxicity studies were also conducted in term of leaching of heavy metals by standard techniques (USEPA using TCLP). Properties of the prepared paint's film in terms of hardness, adhesion, resistance to abrasion, resistance to impact, resistance to corrosion (under humidity and salt fog), etc. were evaluated and compared with a similar formulation of conventional extender and found satisfactory. Results from the experiments indicated that developed extender is environmentally clean and cost-effective.


Retracted: Feature Extraction of Athlete's Post-Match Psychological and Emotional Changes Based on Deep Learning.
[This retracts the article DOI: 10.1155/2022/2995205.].


The cortisol response to hypobaric hypoxia at rest and post-exercise.
High altitude exposure normally leads to a marked natriuresis and diuresis. Acute mountain sickness is often associated with fluid retention, to which an elevated cortisol may contribute. Most investigators report a rise in resting cortisol with ascent, but little data exist regarding the cortisol response to a day trekking. We therefore measured salivary cortisol during ascent to > 5000 m in a cohort of between 42-45 subjects following a 6-h trek (samples taken between 15:30-16:30 h) and between 15-20 subjects at rest (morning samples taken between 08:00-09:00 h). Morning resting cortisol [nmol/l, mean±sd, (range)] was 5.5±2.9 (2.13-13.61) at 1300 m; 4.7±6.8 (1.4-27.02) at 3400 m, and significantly (p=0.002) rose between 4270 m [3.5±2.1 (1.4-8.34)] and 5150 m [14.5±30.3 (1.9-123.1)]. Post-exercise cortisol [nmol/l, mean±sd, (range)] dropped between 3400 m [7±6 (1.5-33.3)] and 4270 m [4.2±4.8 (1.4-29.5)] (p=0.001) followed by a significant rise in post-exercise cortisol between 4270 m [4.2±4.8 (1.4-29.5)] and 5 150 m [9.2±10.2 (1.4-61.3)] (p<0.001). There were no significant associations between severity of acute mountain sickness and cortisol levels. There was a significant though weak correlation between cortisol post-exercise at 5150 m and oxygen saturation at 5150 m (rho= - 0.451, p=0.004). In conclusion, this is the largest cohort to have their resting and post-exercise cortisol levels ascertained at high altitude. We confirm the previous findings of an elevated resting morning cortisol at > 5000 m, but present the novel finding that the cortisol response to a day trekking at HA appears suppressed at 4270 m.


Epithelioid and spindle cell rhabdomyosarcoma with EWSR1::TFCP2 fusion mimicking metastatic lung cancer: A case report and literature review.
Rhabdomyosarcoma (RMS) with EWSR1/FUS::TFCP2 fusion is an emerging, molecularly defined, rare subtype of RMS. It can affect patients in a wide age range and follows an aggressive clinical course according to the reported cases. Due to its unusual clinical and pathohistological features, with a typical intraosseous presentation and common cytokeratin expression, the diagnosis is challenging, and metastatic undifferentiated/sarcomatoid carcinoma can be an important differential diagnosis. We report here a case of a 55-year-old woman with an RMS with EWSR1::TFCP2 fusion mimicking metastatic lung cancer in view of the clinical and microscopic presentation. However, further molecular workup, including RNA sequencing, led to the proper diagnosis. Although these tumors are rare, knowledge of their unique features is essential for correct diagnosis as a basis for clinical management and optimization of therapeutic approaches.


Biosynthesis, characterization and biomedical potential of Arthrospira indica SOSA-4 mediated SeNPs.
The use of aqueous cyanobacterial extracts for selenium nanoparticle (SeNP) synthesis is considered green, cost-effective, and eco-friendly technology that is more advanced than physical and chemical methods. In the current study, an aqueous extract of Arthrospira indica SOSA-4 was used as a reducing and stabilizing agent for the green synthesis of SeNPs. The UV-Visible absorption spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, X-Ray diffraction, Raman spectroscopy, Atomic force microscopy (AFM), Scanning electron microscopy-Energy Dispersive X-Ray spectroscopy(SEM-EDX), and Transmission electron microscopy (TEM) were performed to characterize the biosynthesized SeNPs. Gas chromatography-Mass spectrometry (GC-MS) was also performed to know the composition of the cyanobacterial extract. SEM, TEM, and AFM showed the average size of SeNPs to be 8.5 nm, 9 nm, and 8.7 nm respectively. FT-IR analysis demonstrated the presence of functional groups on the SeNPs that acted as stabilizing agents. XRD pattern and Raman spectroscopy showed the amorphous nature of SeNPs. Synthesized SeNPs showed significant antioxidant activity in DPPH, FRAP, SOR, and ABTS assay. SeNPs showed good anti-microbial activity against Staphylococcus aureus, Escherichia coli, Candida albicans, Candida glabrata, and Candida tropicalis and good anti-cancer activity in MTT assay, Trypan assay, and Flow cytometry analysis against MCF-7, SiHa, and SW480 cell lines. Non-toxicity of SeNPs against normal cell line (HEK-293) was an additional property that affirmed its potential as a bio-compatible nanomaterial.


Tandemly repeated DNA sequences from Xenopus laevis. I. Studies on sequence organization and variation in satellite 1 DNA (741 base-pair repeat).
A family of tandemly repeated sequences, having a basic repeating unit of 741 base-pairs, has been identified in Xenopus laevis DNA and designated satellite 1. Apart from its rather long repeat unit, the characteristics of this DNA appear to be quite similar to those of complex satellite DNAs from other organisms. The nucleotide sequence of a cloned repeat unit shows no evidence of simpler internal repeats, and there is no obvious suggestion of reasonable RNA- or protein-coding regions. Transcripts homologous to this DNA could not be demonstrated in liver, embryo or oocyte RNAs. By blot-hybridization, satellite 1 has been shown to exist in the genome chiefly as tandem repeats of the 741 base-pair sequence. However, there are a number of repeats that differ from the normal sequence (as judged by loss or gain of restriction sites) and some that differ in length. Similar variants are often, but not always, clustered. Characterization of genomic clones of this satellite has confirmed the tandem organization and clustering of variants. The nature of some variants has been elucidated in more detail. Some regions of the basic repeat seem to be more prone to variation than are others.


Analysis of the structure, evolution, and expression of CD24, an important regulator of cell fate.
CD24 is a small, glycophosphatidylinositol-anchored cell surface receptor, expressed in a variety of cells types and tissues. CD24 gene and protein expression is highly dynamic in response to cellular differentiation and stimulation in a cell-specific manner. Furthermore, CD24 interacts with a diverse collection of ligands, including cell adhesion molecules such as P-selectin, and the immune-associated siglec family of transmembrane proteins. While much is known regarding the biological roles of CD24 in regulating cell survival, death and differentiation, little is known about the evolution and organization of CD24 across species or the relationship between CD24 expression and its known ligands. We analyzed the organization and evolution of the CD24 gene from 56 mammalian, avian and reptilian species. We further examined the mRNA expression of CD24 and its known ligands in Mus musculus in immune cells, immunologically privileged tissues, developing brain, and developing and regenerating liver. CD24 arose prior to the reptilian-avian divergence and is conserved across many mammalian species, although we were unable to identify CD24 in marsupials or monotremes. The CD24 genomic structure is diverse between and within species, with varying numbers of exons, introns, and the presence of untranslated regions. Of note, we found no obvious criteria distinguishing CD24 genes from those annotated as CD24-like. The expression of CD24 is similarly complex, with immune cells showing dynamic changes in mRNA levels during development, while immunologically privileged and developing tissues show a high, static expression level that decreases in mature tissues. Furthermore, the expression of CD24 correlated with some but not all of its known ligands in a tissues-specific manner, suggesting that novel ligands have yet to be identified and that cell-specific ligand expression can influence CD24 function. We find that CD24 arose prior to the divergence of reptiles, birds and mammals. Furthermore, the most highly conserved areas of the protein are the amino acids which can be glycosylated. We also find that CD24 expression is highly tissue-specific and in many cases, not well conserved with known CD24 ligands, suggesting yet-unknown CD24-ligand interactions. Together, these data are a valuable resource for furthering studies in CD24 biology.


Prediction of effective Lens position using anterior segment optical coherence tomography in Chinese subjects with angle closure.
To assess the accuracy of biometric parameters measured by anterior segment optical coherence tomography (AS-OCT) and partial coherence interferometry (PCI) in prediction of effective lens position (ELP) compared with previous formulas in PACG patients. 121 PACG eyes were randomly divided into training set (85 eyes) and validation set (36 eyes) with same procedure including AS-OCT, PCI, phacoemulsification and IOL implantation surgery. Preoperative anterior chamber depth (pre-ACD), scleral spur depth (SSD), scleral spur width (SSW), lens vault (LV) and cornea thickness (CT) were measured from AS-OCT image. Axial length (AL) and corneal power (K) were measured by PCI. All the 7 parameters were analyzed by multiple linear regression in training set and a statistic regression formula was developed. In validation set, one-way ANOVA was applied to compare the new regression formula with Sanders-Retzlaff-Kraff theoretic (SRK/T), Holladay 1, Haigis, and a regression formula developed in previous study. The coefficient of determination (R2) of different parameter combinations are 0.19 (pre-ACD, AL), 0.25 (AL, K) and 0.49 (SSD, AL, SSW) in training set. In validation set, the correlation between predicted and measured ELP are: new formula (R2 = 0.50, P = 0.9947) Holladay 1 (R2 = 0.12, P < 0.0001), SRK/T (R2 = 0.11, P < 0.0001) and Haigis (R2 = 0.06, P < 0.0001). Among 7 tested parameters, pre-ACD contribute little in ELP prediction. Formula consist of SSD, AL and SSW showed better accuracy than other formulas tested.


A hybrid microcrystalline cellulose/metal-organic framework for dispersive solid phase microextraction of selected pharmaceuticals: A proof-of-concept.
Solid phase microextraction (SPME) is considered simple, ecofriendly, sustainable, cost-effective and timesaving sample preparation mode in comparison with other sample preparation procedures. The researchers always try to develop new sorbents with higher surface area in comparison with other conventional sorbents aiming to enhance the extraction efficiency. In this work for the first time, a comparative study was performed between Ca-BTC MOF (1,3,5-benzenetricarboxylic acid, BTC; metal-organic framework, MOF) and a hybrid Ca-BTC-MCC MOF (microcrystalline cellulose, MCC) by using as model compounds seven drugs with different physicochemical properties. The evaluation of the extraction efficiency of both sorbents were obtained by means of an HPLC/DAD instrument configuration in reversed phase mode under isocratic elution mode. The results indicate that Ca-BTC MOF showed superior extraction efficiency than Ca-BTC-MCC MOF in the case of all analytes except nirmatrelvir and ritonavir. The results highlight that not only the surface area of adsorbents controlled the adsorption capacity, but also other factors have a role in extraction efficiency including morphology of adsorbent and physico-chemical properties of the analytes. It is worth mentioning that this is the first time that a comparative study was performed between Ca-BTC MOF and Ca-BTC-MCC MOF hybrid material.


Screening for Candidate Genes Associated with Biocontrol Mechanisms of Bacillus pumilus DX01 Using Tn5 Transposon Mutagenesis and a 2-DE-Based Comparative Proteomic Analysis.
A total of 1467 mutants of the biocontrol bacterium Bacillus pumilus DX01 were obtained by Tn5 insertional mutagenesis and subjected to the determination of antagonistic capabilities. Compared with the wild-type strain DX01, the mutant M25 was identified to have the most significant reduction in antagonistic capability against the phytopathogen Bipolaris maydis and extracellular proteinase activity. The integration site of the exogenous T-DNA in the genome of mutant M25 was revealed in the coding region of malony CoA-ACP transacylase (MCAT) gene (mcat), which belongs to a polyketide synthase (PKS) gene cluster, DX01pks of B. pumilus DX01. Furthermore, the whole DX01pks gene cluster was cloned using Illumina Solexa sequencing technology, and it has a modular framework different from the other two gene clusters involved in polyketide synthesis in B. amyloliquefaciens FZB42 (pks1) and B. subtilis 168 (pksX). Finally, in order to gain more insights into the molecular mechanisms of biocontrol of B. pumilus DX01, the changes in the relative level of expression of total proteins between the original strain DX01 and the mutant M25 were detected by 2-DE-based proteomic analysis. A total of twenty differentially expressed proteins were identified upon the mcat gene transposition mutagenesis. Of these proteins, seven proteins were up-regulated in expression level and the other proteins were down-regulated. Taken together, the results in this study showed that Tn5 transposon mutagenesis of B. pumilus DX01 can lead to a significant change of antiphytopathogen ability, and the DX01pks gene cluster possibly play a potential role in the biocontrol processes of this bacterium.


Identification and characterization of N-acetylglucosamine-6-O-sulfate-specific beta1,4-galactosyltransferase in human colorectal mucosa.
6-Sulfo-sialyl Lewis X structure is attributable to recognition between lymphocytes and high endothelial venules. However, the biosynthetic pathway still remains unclear. We found that a beta-galactosyltransferase (betaGalT) in human colorectal mucosa preferentially acts on GlcNAc-6-O-sulfate (6S-GN). 6S-GN:beta4GalT was partially purified by UDP-hexanolamine-Sepharose and asialo-agalacto-ovomucin-Sepharose chromatographies. The optimum pH of this enzyme was found to be 6.5-7.5 and the Michaelis constants for 6S-GN and UDP-Gal were 0.43 mM and 16 microM, respectively. The enzymatic activity was dependent on divalent cations and the substrate specificity was not affected by alpha-lactalbumin. This is the first demonstration of the occurrence of 6S-GN:beta4GalT.


PTEN inhibitor attenuates cardiac fibrosis by regulating the M2 macrophage phenotype via the PI3K/AKT/TGF-β/Smad 2/3 signaling pathway.
Cardiac fibrosis is a key feature of hypertensive cardiac remodeling. In response to microenvironmental stimuli, phenotypic and functional changes in macrophages are considered important determinants of cardiac fibrosis attenuation. VO-OHpic, a phosphatase and tension homolog of chromosome 10 (PTEN) inhibitor, has been demonstrated to be cardioprotective in cardiac remodeling. However, whether VO-OHpic can improve cardiac fibrosis and macrophage polarization remains elusive. The interaction between VO-OHpic and the macrophage phenotype to attenuate cardiac fibrosis was studied in both spontaneously hypertensive rats in vivo and an Ang II-induced hypertension model in vitro. In vitro experiments showed that VO-OHpic promoted M2 macrophage polarization and markedly inhibited proinflammatory M1 macrophages, while VO-OHpic treatment of protein kinase B (AKT)-knockdown/LY294002 (a PI3K inhibitor) macrophages exerted a reduced effect. In a coculture system, culturing cardiac fibroblasts with VO-OHpic-treated macrophages led to significant suppression of proliferation, fibrotic marker expression, and transforming growth factor (TGF)-β and Smad 2/3 protein expression. Taken together, VO-OHpic mediated a fibro-protective effect and increased M2 macrophage polarization via the phosphatidylinositol 3-kinase (PI3K)/AKT/TGF-β/Smad2/3 pathway.


Melanocytic nevi, dysplastic nevi, and malignant melanoma in children from melanoma-prone families.
Melanocytic nevi, particularly dysplastic nevi (DN), are important markers of increased risk of malignant melanoma in adults, but little is known about their prevalence and relation to melanoma in children. Our purpose was to define the prevalence of DN, number of nevi, and their relation to the risk of melanoma in children younger than 20 years of age from melanoma-prone families. One hundred twenty-five persons younger than 20 years of age, from 23 melanoma-prone families, underwent clinical evaluation with nevus counts, photography, and biopsy of suspected melanocytic lesions and were observed for development of DN and melanoma. In melanoma-prone families, 37% of children had DN. The patients were divided into four categories: those with melanoma, DN (without melanoma), indeterminant (largely because of age at examination), and unaffected. The risk of melanoma was assessed by nevus number and presence of DN. High nevus number was strongly correlated with the presence of DN. The risk of the development of melanoma in children from melanoma-prone families appeared most related to the presence of DN (relative risk, 45; 95% confidence intervals, 2.6-786.4) and started at an early age. Of note, all children in whom melanoma developed had DN. Family history of melanoma and the presence of DN defines children with a high risk for melanoma developing at an early age.


Ryanodine receptor leak triggers fiber Ca2+ redistribution to preserve force and elevate basal metabolism in skeletal muscle.
Muscle contraction depends on tightly regulated Ca2+ release. Aberrant Ca2+ leak through ryanodine receptor 1 (RyR1) on the sarcoplasmic reticulum (SR) membrane can lead to heatstroke and malignant hyperthermia (MH) susceptibility, as well as severe myopathy. However, the mechanism by which Ca2+ leak drives these pathologies is unknown. Here, we investigate the effects of four mouse genotypes with increasingly severe RyR1 leak in skeletal muscle fibers. We find that RyR1 Ca2+ leak initiates a cascade of events that cause precise redistribution of Ca2+ among the SR, cytoplasm, and mitochondria through altering the Ca2+ permeability of the transverse tubular system membrane. This redistribution of Ca2+ allows mice with moderate RyR1 leak to maintain normal function; however, severe RyR1 leak with RYR1 mutations reduces the capacity to generate force. Our results reveal the mechanism underlying force preservation, increased ATP metabolism, and susceptibility to MH in individuals with gain-of-function RYR1 mutations.


WT1 in disease: shifting the epithelial-mesenchymal balance.
WT1 is a versatile gene that controls transitions between the mesenchymal and epithelial state of cells in a tissue-context dependent manner. As such, WT1 is indispensable for normal development of many organs and tissues. Uncontrolled epithelial to mesenchymal transition (EMT) is a hallmark of a diverse array of pathologies and disturbance of mesenchymal to epithelial transition (MET) has been associated with a number of developmental abnormalities. It is therefore not surprising that WT1 has been linked to many of these. Here we review the role of WT1 in proper control of the mesenchymal-epithelial balance of cells and discuss how far these roles can explain the role of WT1 in a variety of disease states.


Changes in the rate of photosynthesis accompanying the yield increase in wheat cultivars released in the past 50 years.
Photosynthetic rates in different development stages were carefully investigated in 18 cultivars of winter wheat released in the period between 1945 and 1995 in the area of Beijing, China. During this period, the recorded grain yield has increased eightfold. However, when those cultivars were planted and managed in the same environment, the difference was reduced to only 36%, indicating that agronomic practices are the most important factors for grain yield. Agronomic features have changed greatly in the past 50 years, through increasing the harvest index (R2 = 0.89, P < 0.05), shortening plant height (R2 = 0.77, P < 0.05) and slightly increasing flag leaf areas (R2 = 0.45, P < 0.05), which is mostly in agreement with many other researchers. In contrast to many reports, however, this study found a genetic increase in the rate of photosynthesis per unit leaf area. From the mid-stem elongation to soft dough stages, the average photosynthetic rates at saturated photosynthetic photon flux density (P(sat)) increased by 44%. In the process, the stomatal conductance (g(s)) also increased by 122%. Grain yield was positively related to the mean values of P(sat) (R2 = 0.61, P < 0.01) and g(s) (R2 = 0.67, P < 0.01) in the six development stages. Our experiment may suggest that increase in grain yield was associated with the elevation of leaf photosynthetic rate and stomatal conductance over the past 50 years.


Intratumoural mRNA expression of genes from the oestradiol metabolic pathway and clinical and histopathological parameters of breast cancer.
The expression of the oestrogen receptor (ER) is one of the more important clinical parameters of breast cancer. However, the relationship between the ER and its ligand, oestradiol, and the enzymes that synthesise it are not well understood. The expression of mRNA transcripts of members of the oestradiol metabolic and signalling pathways including the ER was studied in detail. mRNA transcripts for aromatase (CYP19), 17-beta-hydroxysteroid dehydrogenase I, 17-beta-hydroxysteroid dehydrogenase II, ERalpha, ERbeta, steroid sulfatase (STS), oestradiol sulfotransferase (EST), cyclin D1 (CYCLD1) and ERBB2 were fluorometrically quantified by competitive RT-PCR using an internal standard in 155 breast carcinomas. In addition, the transcripts of CYP19 were analysed for alternative splicing/usage of exon 1 and an alternative poly A tail. A great variability of expression was observed, ranging from 0 to 2376 amol/mg RNA. The highest levels were observed for STS and EST, and the lowest levels (close to zero) were observed for the 17-beta-hydroxysteroid dehydrogenase isoenzymes. The levels of mRNA expression were analysed with respect to clinical and histopathological parameters as well as for disease-free survival. High correlation of the mRNA expression of STS, EST and 17-beta-hydroxysteroid dehydrogenase in the tumours suggested a common regulation, possibly by their common metabolite (oestradiol). Hierarchical clustering analysis in the 155 patients resulted in two main clusters, representing the ERalpha-negative and ERalpha-positive breast cancer cases. The mRNA expression of the oestradiol metabolising enzymes did not follow the expression of the ERalpha in all cases, leading to the formation of several subclasses of tumours. Patients with no expression of CYP19 and patients with high levels of expression of STS had significantly shorter disease-free survival time (P > 0.0005 and P < 0.03, respectively). Expression of ERbeta mRNA was a better prognostic factor than that of ERalpha in this material. Our results indicate the importance of CYP19 and the enzymes regulating the oestrone sulfate metabolism as factors of disease-free survival in breast cancer, in addition to the well-known factors ER and ERBB2.


DRP1 upregulation promotes pancreatic cancer growth and metastasis through increased aerobic glycolysis.
Mitochondrial shape is dynamically changed by fusion and fission processes in cells, and dysfunction of this process has become one of the emerging hallmarks of cancer. However, the expression patterns and biological effects of mitochondrial fission and fusion proteins in pancreatic cancer (PC) are still unclear. The expressions of mitochondrial fission and fusion proteins were first evaluated by quantitative reverse transcription polymerase chain reaction and western blot analysis in both PC cell lines and tissue samples. In addition, the biologic functions of the differentially expressed proteins in PC cell growth and metastasis both in vitro and in vivo and their potential underlying mechanisms were systematically explored. We first found that DRP1 was substantially upregulated in PC cell lines and tissue samples mainly due to the downregulation of miR-29a, which contributed to the poor survival of PC patients. DRP1 promoted the growth and metastasis of PC cells both in vitro and in vivo by inducing G1-S cell cycle transition and matrix metalloproteinase 2 secretion. Mechanistic investigations revealed that increased DRP1 upregulation-mediated mitochondrial fission and subsequently enhanced aerobic glycolysis were involved in the promotion of growth and metastasis by DRP1 in PC cells. Our findings demonstrate that mitochondrial fusion protein DRP1 plays a critical oncogenic role in PC cells by enhancing aerobic glycolysis, which could serve as a novel therapeutic target for PC treatment.


Presynaptic calcium channels: pharmacology and regulation.
Voltage-dependent Ca2+ channels are considered as molecular trigger elements for signal transmission at chemical synapses. Due to their central role in this fundamental process, function and pharmacology of presynaptic Ca2+ channels have recently been the subject of extensive exploration employing various experimental techniques. Several lines of evidence indicate that, at nerve terminals in higher vertebrates, the evoked influx of Ca2+ -ions is mainly mediated by Ca2+ channels of the P-type. The stringent regulation of presynaptic Ca2+ channels is supposed to be involved in fine-tuning the efficiency of synaptic transmission. Intrinsic control mechanisms, such as voltage- or Ca(2+)-dependent inactivation, or modulation of channel activity, either by G-proteins directly or via phosphorylation by protein kinases, may be of particular functional importance.


Priapism as a sequela of chlorpromazine therapy.
A patient developed priapism following a single intramuscular injection of chlorpromazine for hiccups. Conservative management was unsuccessful. During surgical intervention on the 15th hospital day, the corpora cavernosa of the turgid penis was irrigated and aspirated with heparinized saline using No14 gauge needles. The patient did well postoperatively. The physiology of the erection and the pathophysiology of priapism are discussed.


A systematic review and meta-analysis of immediate postpartal intrauterine contraceptive device use in Ethiopia.
Immediate postpartum intrauterine device (IPPIUCD) use remains too low in Ethiopia and there are high levels of unmet need for IPPIUCD. This systematic review and meta-analysis investigates individual studies conducted in Ethiopia on IPPIUCD use and influencing factors. Extensive database searching was done using Google Scholar, PubMed, Scopes, Cochrane library, Web of Science, and Science Direct search engines. Data were extracted and analyzed using Cochrane review manager version 5.4.1. A random-effects model with a 95% confidence interval (CI) was used to estimate the pooled magnitude of IPPIUCD use. Forest plot was used to estimate the pooled IPPIUCD use and inverse variance was used to identify the presence of heterogeneity. Publication bias was assessed by using funnel plots and Egger's statistical tests. This study showed that the pooled use of IPPIUCD in Ethiopia was 8.37% (95% CI: 4.32, 16.21). Those who had heard about IPPIUCD were 4.2 times (OR = 4.2, 95% CI: 1.51, 11.68) more likely, had birth interval >3 years were 3.90 times (OR = 3.90, 95% CI: 1.68, 9.05) more likely, had good knowledge were 4.44 times (OR = 4.44, 95% CI: 2.26, 8.76) more likely and counseled clients were 3.99 times (OR = 3.99, 95% CI: 1.28, 12.37) more likely to use IPPIUCD; and the odds of using IPPIUCD was 45% (OR = 0.55, 95% CI: 0.40, 0.77) less likely among aged 15-24 years old to use IPPIUCD. IPPIUCD use in Ethiopia was low. Age category, ever heard about IPPIUCD, level of knowledge, birth interval and being counseled about IPPIUCD were statistically significant factors influencing IPPIUCD use.


Outcomes of Severe Seronegative Hepatitis-associated Aplastic Anemia: A Pediatric Case Series.
Hepatitis-associated aplastic anemia (HAAA) is a potentially life-threatening diagnosis without clear treatment guidelines. The goal of the study was to characterize the presentation, evaluation, histopathology, and outcomes of therapy in children with HAAA to guide future research and to develop standardized care guidelines for this rare disease. Retrospective chart review of 4 patients with HAAA who presented to Children's Hospital Colorado between 2016 and 2019 was conducted. Patient presentation, evaluation, bone marrow and liver pathology, interventions, and clinical course were collected. Immunohistochemistry of liver biopsies was performed. We treated 4 patients with HAAA without liver failure. All had evidence of systemic hyperinflammation and CD8+ T cell predominant liver tissue infiltration. One had a genetic mutation predisposing him to immune-mediated disease, but all other genetic testing was negative. In 3 of the 4 patients, hepatitis was poorly responsive to standard therapy with steroids, azathioprine, or tacrolimus; however, sustained biochemical remission of hepatitis was induced after more aggressive immunosuppressive therapies including Anti-Thymocyte Globulin (ATG) at standard immunosuppressive therapy (IST) dosing for severe Aplastic Anemia (sAA). Two patients underwent hematopoietic stem cell transplant (HSCT); 1 as first line therapy and 1 for refractory sAA. We found that ATG-based IST induced remission of hepatitis in patients with steroid-refractory HAAA. This is also an appropriate initial treatment for severe Aplastic Anemia, though may not prevent the need for HSCT. We propose that equine ATG based IST at standard dosing regimen for sAA is a therapy that in select cases can be considered early on in the treatment course and could lead to a sustained remission of both hepatitis and sAA. This should be considered in collaboration with a pediatric hematologist.


Posterior segment manifestations of HIV/AIDS.
The ocular posterior segment manifestations of AIDS may be divided into four categories: retinal vasculopathy, unusual malignancies, neuro-ophthalmologic abnormalities, and opportunistic infections. Microvasculopathy is the most common manifestation. Opportunistic infections, particularly cytomegalovirus retinitis and progressive outer retinal necrosis, are the most likely to result in visual loss due to infection or subsequent retinal detachment. Diagnosis and treatment are guided by the particular conditions and immune status of the patient.


"Triple infections" (fungal, bacterial and viral) in immunosuppressed renal transplant recipients.
In a period of 5 years, 8 out of 77 renal transplant patients showed simultaneous fungal, bacterial and viral infections. Candida albicans was found in all cases. The most severe bacterial complications were infections with Klebsiella, Pseudomonas and Staphylococcus aureus. Cytomegalovirus, persistent HBsAg positive hepatitis, herpes zoster, and herpes simplex infections were also found. Seven patients died of bacterial superinfection and miliary tuberculosis. The data presented show that "triple infections" are associated with high mortality and that miliary tuberculosis occurred frequently in immunosuppressed renal transplant recipients.


Targeting cancer stem cells as therapeutic approach in the treatment of colorectal cancer.
Colorectal cancer is one of the most common cancers globally. A large portion of colorectal cancer patients who are treated with conventional chemotherapy eventually develop local recurrence or metastases. The failure of a complete cure in colorectal cancer patients may be related to the lack of complete eradication of cancer stem cells when using conventional therapy. Colorectal cancer stem cells comprise a small population of tumor cells that possess the properties of rapid proliferation and differentiation. The colorectal cancer stem cells are also phenotypically and molecularly distinct, and resistant to conventional chemo-radiotherapy. Therefore, it is important to identify approaches in combination with conventional therapy for targeting and eradicating cancer cells. The aim of this review was to summarize the main findings of recent studies on targeting colorectal cancer stem cells as a novel therapeutic approach in colorectal cancer treatment.


Determinants of the uptake of medicines use reviews (MURs) by community pharmacies in England: a multi-method study.
To explore and identify the key determinants influencing the uptake of medicines use reviews (MURs), a new community pharmacy service in England. Survey of all primary care organisations (PCOs) in England (n=303, response rate=74%) and case study investigations of 10 PCOs, involving interviews with a purposive sample of 43 key stakeholders, including PCO, Local Pharmaceutical Committee and community pharmacy representatives. National data on MUR activity were also analysed and multiple linear regression was used to test determinants of MUR uptake. The ownership category of the pharmacy was shown to be the most significant determinant of MUR uptake. Rates of MUR provision by multiple pharmacies were almost twice that of independent pharmacies. Interview data corroborated this finding, suggesting that organisational pressure within multiple pharmacies was driving forward MUR activity in some PCOs. Interviewees expressed concern about this quantity driven approach. The PCO survey respondents perceived the greatest barrier to MUR implementation to be a lack of support from general practitioners (GPs). Interviewees reported a lack of communication about MURs between community pharmacists and GPs. The findings suggest that the organisational setting of the pharmacy is an important factor influencing the uptake of MURs. There is also a need for greater communication and collaboration with GPs regarding the MUR service.


Identification of the social and cognitive processes underlying human cumulative culture.
The remarkable ecological and demographic success of humanity is largely attributed to our capacity for cumulative culture, with knowledge and technology accumulating over time, yet the social and cognitive capabilities that have enabled cumulative culture remain unclear. In a comparative study of sequential problem solving, we provided groups of capuchin monkeys, chimpanzees, and children with an experimental puzzlebox that could be solved in three stages to retrieve rewards of increasing desirability. The success of the children, but not of the chimpanzees or capuchins, in reaching higher-level solutions was strongly associated with a package of sociocognitive processes-including teaching through verbal instruction, imitation, and prosociality-that were observed only in the children and covaried with performance.


Post-discharge oral nutritional supplements with dietary advice in patients at nutritional risk after surgery for gastric cancer: A randomized clinical trial.
Malnutrition frequently occurs and deteriorates in patients after surgery for gastric cancer, especially after hospital discharge, which has been consistently associated with negative outcomes. However, information regarding the impact of post-discharge nutritional interventions is poorly described. The aim of this study was thus to evaluate the impact of post-discharge oral nutritional supplements (ONS) with dietary advice compared with dietary advice alone on nutritional outcomes, including body mass index (BMI) and skeletal muscle index (SMI), sarcopenia prevalence, chemotherapy tolerance, the 90-day readmission rate, and quality of life in patients at nutritional risk after surgery for gastric cancer. Three hundred and fifty-three patients who underwent surgery for gastric cancer and were at nutritional risk (Nutritional Risk Screening 2002 [NRS 2002] score ≥3 points) in our institution were randomly assigned to receive either ONS with dietary advice or dietary advice alone (control) for 3 months after discharge. The primary endpoints were nutritional outcomes and sarcopenia prevalence; the secondary endpoints included chemotherapy tolerance, the 90-day readmission rate, and quality of life. Three hundred and thirty-seven patients completed the study and were included in the analyses, consisting of 171 in the ONS group and 166 in the control group. The average daily intake of ONS in the intervention group was 370 mL. After 3 months of the intervention, the patients who received ONS and dietary advice had significantly less weight loss and higher BMI and SMI than those given dietary advice alone (P < 0.05). The incidence of sarcopenia was significantly lower in the ONS group than in the control group (P < 0.05). Similar number of patients in the two groups underwent postoperative chemotherapy, but the patients who received ONS and dietary advice had significantly less chemotherapy modifications, including delay, dose reduction, or termination (P < 0.05). The two groups had no significant differences in the 90-day readmission rate (P > 0.05). Regarding the quality of life, the patients who received ONS and dietary advice reported significantly less fatigue and appetite loss than those given dietary advice alone (P < 0.05), but the two groups showed no significant differences in the other outcomes (P > 0.05). Post-discharge ONS with dietary advice in patients at nutritional risk after surgery for gastric cancer improved nutritional outcomes, skeletal muscle maintenance, chemotherapy tolerance and some quality of life variables. These findings strongly support the concept of the introduction of post-discharge ONS with dietary advice to this patient cohort.


Technique for thrombectomy of reversed saphenous vein arterial bypass grafts.
A technique for thrombectomy of saphenous vein arterial bypass grafts is described. The technique includes visual inspection of both anastomoses and avoids unnecessary trauma to the graft secondary to repeated attempts at passing a balloon catheter against the direction of the venous valves.


Noble gas-transition metal complexes: coordination of ScO+ by multiple Ar, Kr, and Xe atoms in noble gas matrixes.
The combination of matrix isolation infrared spectroscopic and density functional calculation results provides strong evidence that the transition metal monoxide cation, ScO+, coordinates five noble gas atoms in forming the [ScO(Ng)5]+ (Ng = Ar, Kr, or Xe) complexes in noble gas matrixes.


Continuously Varying Critical Exponents Beyond Weak Universality.
Renormalization group theory does not restrict the form of continuous variation of critical exponents which occurs in presence of a marginal operator. However, the continuous variation of critical exponents, observed in different contexts, usually follows a weak universality scenario where some of the exponents (e.g., β, γ, ν) vary keeping others (e.g., δ, η) fixed. Here we report ferromagnetic phase transition in (Sm1-yNdy)0.52Sr0.48MnO3 (0.5 ≤ y ≤ 1) single crystals where all three exponents β, γ, δ vary with Nd concentration y. Such a variation clearly violates both universality and weak universality hypothesis. We propose a new scaling theory that explains the present experimental results, reduces to the weak universality as a special case, and provides a generic route leading to continuous variation of critical exponents and multi-criticality.


Concatemerization and packaging of bacteriophage T7 DNA in vitro: determination of the concatemers' length and appearance kinetics by use of rotating gel electrophoresis.
During its morphogenesis both in intact infected cells (in vivo) and in lysates of infected cells (in vitro), bacteriophage T7 forms end-to-end concatemers of its mature DNA, a linear, nonpermuted, terminally repetitious DNA. During morphogenesis, in vivo T7 concatemers are packaged in preformed capsids and cut to mature size. In the present study the lengths and appearance kinetics of concatemers formed in vitro from mature T7 DNA have been determined. The following procedures are used here for the first time: (a) 20-35% efficient in vitro concatemerization and packaging of T7 DNA; the mixture used for packaging contained two lysates that together had all T7 gene products, and (b) fractionation of concatemers by rotating gel electrophorsis (RGE), which improves the resolution by length of concatemer-length DNA. Concatemerization at 30 degrees was so fast that some other process must be rate limiting for packaging. The concatemers formed were linear and joined left-end to right-end by complementary base pairing, not by blunt-end ligation. Concatemers formed at 30 degrees were reconverted to mature DNA by packaging in vitro. Reducing the temperature to 0 degrees both slowed concatemerization to the time scale (minutes) needed for control of the extent of concatemerization and reduced packaging to insignificant levels, thereby also uncoupling packaging from concatemerization. At both 30 degrees and 0 degrees bands of discrete-length concatemers were observed by RGE. The lengths were n times the length of mature T7 DNA; n was found to be any integer from 2 to 15. The bands were stronger at 0 degrees than they were at 30 degrees in comparison to a background of heterogeneous DNA. No evidence for the favoring of any value of n was found. In addition, it was found by two-dimensional agarose gel electrophoresis that a comparatively small amount of circular DNA was produced in vitro.


Purification and properties of a protein that binds to the C-terminal coding region of human c-myc mRNA.
The short half-life of c-myc mRNA is influenced by sequences in the 3'-untranslated region and the C-terminal part of the coding region. In cell-free extracts, a polysomal protein binds to RNA corresponding to the coding region stability determinant. This and other observations suggest that the protein is bound to polysome-associated c-myc mRNA and protects the mRNA from a ribosome-associated endoribonuclease (Bernstein, P.L., Herrick, D.J., Prokipcak, R.D., and Ross, J. (1992) Genes & Dev. 6, 642-654). Here, we describe a four-step purification of the binding protein: solubilization from ribosomes, ammonium sulfate precipitation, RNA affinity chromatography, and reverse-phase high performance liquid chromatography. The 70-kDa protein can be renatured from solutions containing sodium dodecyl sulfate or organic solvents, greatly facilitating its purification. Protein binding to c-myc coding region RNA is blocked by diamide and N-ethylmaleimide, indicating a requirement for sulfhydryl groups. The protein also binds to N-myc coding region RNA but with approximately 5-fold lower affinity than to the comparable c-myc region. Excess c-myc competitor RNA induces 8-fold destabilization of c-myc mRNA in cell-free mRNA decay extracts. In contrast, N-myc coding region competitor RNA has no effect on c-myc mRNA half-life. Therefore, the protein we have purified probably affects c-myc mRNA metabolism with high specificity.


Asymmetry in glenohumeral muscle strength of sitting volleyball players: an isokinetic profile of shoulder rotations strength.
Sitting volleyball (SV) is a popular Paralympic game, in which shoulders of SV players are overloaded continuously during play. Nevertheless, shoulder girdle muscle function has never been measured in this population yet. The objective of this study was to investigate the dominant and non-dominant strength profile of male and female elite SV players on a glenohumeral level. A total of 13 Brazilian SV national team players volunteered to participate in this study (male: N.=6, age 32.8±4.1 years, body mass 83.2±19.4 kg, seated height 1.47±0.04 m; female: N.=7, age 32.8±4.1 years, body mass 80.1±17.9 kg, seated height 1.36±0.05 m). A Biodex dynamometer was used to measure internal rotation (IR) and external rotation (ER) of the glenohumeral muscle strength at 60°/s and 180°/s velocities in a concentric/concentric mode. It was observed that male SV players were stronger than the female players at both upper limbs. Concerning the side effects, significant differences were found for IR at 60°/s velocity (F=7.55, P=0.02) and ER at 180°/s velocity (F=5.91, P=0.03), with higher values on the dominant limb in comparison to the non-dominant limb for both the male and female players. There were significant gender differences for the IR inter-limb asymmetry at 180°/s velocity, with a greater asymmetry between both shoulders in the male players compared to the female players, preference for the dominant side in both groups. The SV players have an asymmetrical rotational strength profile. As muscular imbalance is a common injury pattern among overhead players. The findings of this study may assist the SV coaches and physiotherapists for evaluation and training of strength of the ER and IR shoulders of these players.


Pertuzumab, trastuzumab, and docetaxel for Chinese patients with previously untreated HER2-positive locally recurrent or metastatic breast cancer (PUFFIN): a phase III, randomized, double-blind, placebo-controlled study.
The Chinese bridging study PUFFIN (NCT02896855) aimed to assess consistency of efficacy with CLEOPATRA (NCT00567190), investigating pertuzumab with trastuzumab and docetaxel versus placebo, trastuzumab, and docetaxel in patients with previously untreated HER2-positive locally recurrent or metastatic breast cancer. Patients were randomized 1:1, stratified by visceral/non-visceral disease and hormone receptor status. The primary endpoint was investigator-assessed progression-free survival (PFS). Secondary endpoints included objective response rate (in patients with measurable baseline disease), overall survival, and safety. The consistency threshold for PFS (hazard ratio [HR] < 0.81) (maintaining ≥ 50% of the risk reduction determined in CLEOPATRA [HR 0.62]) determined the target sample size (n = 240). Two hundred forty-three patients were randomized. Median PFS was 14.5 months in the pertuzumab arm (95% confidence interval [CI] 12.5, 18.6) and 12.4 months in the placebo arm (95% CI 10.4, 12.7) in the intention-to-treat population (HR: 0.69 [95% CI 0.49, 0.99]). Objective responses were recorded in 83/105 (79.0%) and 67/97 (69.1%) patients, respectively. Grade ≥ 3 adverse events (70.5% and 69.2%, respectively) and serious adverse events (19.7% and 19.2%, respectively) were similar across both arms. No heart failure cases or symptomatic left ventricular ejection fraction declines were reported. PUFFIN met its primary objective. Overall, efficacy data were consistent with CLEOPATRA. Safety was consistent with the known pertuzumab safety profile. PUFFIN adds to the totality of data with pertuzumab in previously untreated HER2-positive locally recurrent or metastatic breast cancer and supports the favorable benefit-risk profile of pertuzumab in Chinese patients TRIAL REGISTRATION: ClinicalTrials.gov, NCT02896855, registered 7 September 2016.


Investigation of myringosclerosis development in different grafting materials after myringo-/tympanoplasty.
This study aims to investigate the prevalence of myringosclerosis in tragal perichondrium (TP) and temporalis fascia (TF) grafts after myringo-/tympanoplasty. Thirty patients, who were diagnosed with non-suppurative chronic otitis media and administered myringo-/tympanoplasty under general anesthesia using TP or TF as grafting material in our clinic between January 2012 and May 2013, were recruited in this retrospective case-control study. Patients were divided into two groups according to used graft material as group 1 (TP) (9 males, 6 females; median age 35.4 years; range 20 to 62 years) and group 2 (TF) (7 males, 8 females; median age 39.8 years; range 19 to 63 years). Occurrence of myringosclerosis on the intact tympanic membrane grafts was evaluated postoperatively by the same surgeon under surgical microscope. Postoperative myringosclerosis was detected in six patients (40%) in TP group and 13 patients (86.7%) in TF group. Prevalence of myringosclerosis on graft materials used in myringo-/tympanoplasty was lower in TP group compared to TF group. We believe that this may be due to different vascular structures of different graft tissues and that the low predisposition of perichondrium towards development of myringosclerosis can be considered as an advantage for this graft. We hope that this study adds a new dimension to etiopathogenesis of myringosclerosis and assists otologists in the prevention and treatment of this lesion.


Multidisciplinary Clinical Approach to Cancer Patients with Immune-Related Adverse Events Induced by Checkpoint Inhibitors.
Immune-oncology is a major breakthrough in cancer treatment and has become the standard of care for a wide variety of solid organ malignancies. Unfortunately, manipulation of the immune system with checkpoint inhibitors may result in an immune-based attack of normal tissues which can lead to treatment discontinuation. These immune-related adverse events (irAEs) are diverse and affect several organs, constituting a new clinical challenge in the management of cancer patients. The complexity of this scenario requires a multidisciplinary approach that allows the early identification, diagnosis and treatment of specific irAE, ruling out other non-related adverse events. Hospital Clinic has a multidisciplinary team seeking to develop a coordinated strategy to facilitate the access of patients with suspected irAEs to specialised care resulting in harmonised management that guarantees the best patient care. The aim of the manuscript was to describe the current evidence on the management of irAEs reflecting a coordinated multidisciplinary approach to face this clinical challenge regardless of the immunotherapy indication.


Natural killer cell stimulatory factor (NKSF) augments natural killer cell and antibody-dependent tumoricidal response against colon carcinoma cell lines.
The therapy of colorectal cancer may be improved by biologic response modifiers that enhance natural killer (NK) cell and antibody-dependent tumoricidal mechanisms. This study examined the effect of a recently discovered cytokine purified from the supernatant of an Ebstein-Barr virus-transformed B-lymphoblastoid cell line (RPMI-8866), natural killer cell stimulatory factor (NKSF), on NK and antibody-dependent cellular cytotoxicity (ADCC) of human colon adenocarcinoma cell lines. Human peripheral blood lymphocytes were cultured for 24 hr in the presence or absence of NKSF (3.6 pM) or interleukin-2 (1 nM). The cultured lymphocytes were analyzed for lytic potential toward chromium-51-labeled colon carcinoma targets SW 1116, 498 LI, and WC 1. ADCC was measured by incubating chromium-51-labeled SW 1116 or WC 1 targets with the monoclonal antibody CO17-1A, an IgG2a antibody reactive with gastrointestinal cancer-associated cell antigen, or control mouse IgG prior to testing NKSF-treated or control PBL effectors in a 6-hr cytotoxicity assay. NKSF significantly enhanced NK cytolysis of colon carcinoma and NK-resistant lymphoma cell lines, and on a molar basis was approximately 300 times more potent than interleukin-2 in generating NK cytotoxicity. Furthermore, NKSF significantly augmented lymphocyte-mediated ADCC against colon carcinoma targets, and the combination of NKSF with the antibody CO17-1A had an additive effect on lymphocyte tumoricidial capacity. Thus, NKSF may have a potential role in the treatment of colon cancer.


Burnout and compassion fatigue among organ donation coordinators: a scoping review protocol.
The objective of this review is to develop a comprehensive description of burnout and compassion fatigue, including risk/protective factors, among organ and tissue donation coordinators worldwide. Research on turnover rates among organ and tissue donation coordinators has shown that job tenure generally lasts less than three years, a possible consequence of burnout and compassion fatigue. Increased turnover rates of organ and tissue donation coordinators have significant impact on the ability of ODOs to optimize organ donation. This current scoping review will help inform understanding of the nature of burnout and compassion fatigue in this population as well as the available strategies for managing it. Articles examining health care professionals working as organ and tissue donation coordinators, with the main concepts of interest being burnout and/or compassion fatigue, will be considered. Articles within the context of organ and tissue donation coordinators in acute care settings at any part of the organ donation process will also be considered for inclusion in this review. Quantitative and qualitative studies, text and opinion papers, unpublished material (eg, unpublished research data, reports, institutional protocols, government documents) provided by researchers in the organ donation field worldwide will also be examined for possible inclusion. The scoping review will be performed in accordance with JBI methodology. Published and unpublished papers from 1980 to the present will be searched. Databases will include MEDLINE (Ovid), CINAHL, Embase, LILACS, PsycINFO, and PTSDpubs, while appropriate gray literature will be searched. Two reviewers will screen the papers according to predefined inclusion and exclusion criteria, extract data for specific variables, and perform descriptive examination.


Association of fish and long-chain n-3 polyunsaturated fatty acid intakes with the occurrence of depressive episodes in middle-aged French men and women.
This study aimed to seek whether habitual fish and seafood or n-3 long-chain PUFA intake could influence the occurrence of depressive episodes. In a subsample from the French SU.VI.MAX cohort, dietary habits have been assessed during the first 2 years of the follow-up (six 24-h records) and declarations of antidepressant prescription, taken as markers of depressive episodes, have been recorded during the 8-year follow-up. Subjects consuming fatty fish or with an intake of long-chain n-3 PUFA higher than 0.10% of energy intake had a significantly lesser risk of any depressive episode and of recurrent depressive episodes, but not of single depressive episode. These associations were stronger in men and in non-smokers. In contrast, smokers eating fatty fish had an increased risk of recurrent depression. These results suggest that a usual intake of fatty fish or long-chain n-3 PUFA may decrease the risk of recurrent depression in non-smokers.


Transplantation of Differentiated Tonsil-Derived Mesenchymal Stem Cells Ameliorates Murine Duchenne Muscular Dystrophy via Autophagy Activation.
Skeletal muscles play many important roles in the human body and any malfunction or disorder of the skeletal muscles can lead to a reduced quality of life. Some skeletal dysfunctions are acquired, such as sarcopenia but others are congenital. Duchenne muscular dystrophy (DMD) is one of the most common forms of hereditary muscular dystrophy and is caused by a deficiency of the protein, Dystrophin. Currently, there is no clear treatment for DMD, there are only methods that can alleviate the symptoms of the disease. Mesenchymal stem cells, including tonsil-derived mesenchymal stem cells (TMSCs) have been shown to differentiate into skeletal muscle cells (TMSC-myocyte) and can be one of the resources for the treatment of DMD. Skeletal muscle cell characteristics of TMSC-myocytes have been confirmed through changes in morphology and expression of skeletal muscle markers such as Myogenin, Myf6, and MYH families after differentiation. Based on these characteristics, TMSC-myocytes have been transplanted into mdx mice, a mouse model of DMD, to investigate whether they can help improve the symptoms of DMD. The red fluorescent protein gene was transduced into TMSC (TMSC-R) for tracking transplanted cells. Prior to transplantation (TP), it was confirmed whether TMSC-R-myocytes had the same differentiation potential as TMSC-myocytes. Increased expression of dystrophin and autophagy markers in the TP group compared with the sham group was confirmed in the gastrocnemius muscle 12 weeks after TP. These results demonstrate muscle regeneration and functional recovery of mdx via autophagy activation following TMSC-myocyte TP.


Report of new variants in PPIL1 underlying type 14 pontocerebellar hypoplasia and their associated phenotypic manifestations in two fetuses.
Mutations in the PPIL1 gene have been linked to type 14 pontocerebellar hypoplasia (PCH14); however, prenatal clinical characteristics of PCH14 caused by mutations in the PPIL1 gene have not been reported. This study reports the first prenatal case of PCH14 diagnosed by whole-exome sequencing (WES). Two fetuses with severe microcephaly and cerebral dysplasia, along with their parents, underwent WES. The effects of the discovered PPIL1 variants on PPIL1 protein function were investigated using bioinformatics tools. WES revealed two compound heterozygous missense mutations in PPIL1, c.376C > G (p.His126Asp) and c.392G > T (p.Arg131Leu), inherited from the mother and father, respectively. The co-segregation of PPIL1 mutations in this family was confirmed using Sanger sequencing, identifying two PCH14-affected fetuses. Bioinformatics analysis revealed that these mutations could disrupt the formation of hydrogen bonds, altering the structural stability of the PPIL1 protein. This study is the first to define the clinical characteristics of PCH14 during pregnancy and reports a novel heterozygous missense variant, expanding the PCH14-related mutational spectrum of PPIL1.


The Intersection of Aging and Lung Transplantation: its Impact on Transplant Evaluation, Outcomes, and Clinical Care.
Older adults (age ≥ 65 years) are the fastest growing age group undergoing lung transplantation. Further, international consensus document for the selection of lung transplant candidates no longer suggest a fixed upper age limit. Although carefully selected older adults can derive great benefit, understanding which older adults will do well after transplant with improved survival and health-related qualiy of life is key to informed decision-making. Herein, we review the epidemiology of aging in lung transplantation and its impact on outcomes, highlight selected physiological measures that may be informative when evaluating and managing older lung transplant patients, and identify directions for future research. In general, listing and transplanting older, sicker patients has contributed to worse clinical outcomes and greater healthcare use. Emerging evidence suggest that measures of physiological age, such as frailty, body composition, and neurocognitive and psychosocial function, may better identify risk for poor transplant outcomes than chronlogical age. The evidence base to inform transplant decision-making and improvements in care for older adults is small but growing. Multipronged efforts at the intersection of aging and lung transplantation are needed to improve the clinical and patient centered outcomes for this large and growing cohort of patients. Future research should focus on identifying novel and ideally modifiable risk factors for poor outcomes specific to older adults, better approaches to measuring physiological aging (e.g., frailty, body composition, neurocognitive and psychosocial function), and the underlying mechanisms of physiological aging. Finally, interventions that can improve clinical and patient centered outcomes for older adults are needed.


Distribution of norepinephrine and dopamine in cerebral cortical areas of the rat.
Concentrations of norepinephrine and dopamine were determined using enzyme isotope assay in 27 microdissected cerebral cortical areas of the rat. A detailed map is presented for microdissection of rat cerebral cortex. Norepinephrine was found in low but still measurable quantities throughout the cortex. Differences between cortical areas are also low. Relatively highest levels were demonstrated in the pyriform, insular and entorhinal cortices. The distribution of dopamine was found to be uneven with a maximal regional difference of 1:24. Concentration of dopamine was in all areas lower than that of norepinephrine. The highest dopamine concentration (2,4 ng/mg protein) was measured in the rostral pyriform cortex but other mesocortical (cingulate, frontal, insular and entorhinal) dopaminergic areas also contained relatively high amounts. Except for the caudal occipital and caudal entorhinal cortices all regions studied contained measurable quantities of dopamine. Its low concentration relative to norepinephrine (below 15%) suggests that in the cortical areas studied dopamine is present as the precursor of norepinephrine.


The thyrotropin releasing hormone analogue, RGH 2202, reverses experimental haemorrhagic shock in rats.
Hypovolemic shock was produced in rats by withdrawing about 50% of the estimated total blood volume. Following mean arterial pressure stabilization in the range of 22-23 mm Hg, the rats were given an i.v. bolus injection of L-6-ketopiperidine-2-carbonyl-L-leucyl-L-proline amide (RGH-2202) to be compared with thyrotropin-releasing hormone (TRH). RGH-2202, administered within 5 min following shock induction, dose dependently improved the mean arterial pressure and survival of the rats. The activity of RGH-2202 was superior to that of TRH after i.v. administration; both drugs showed a lower activity after i.m. administration.


The Effects of Appearance-Based Comments and Non-Appearance-Based Evaluations on Body Dissatisfaction and Disordered Eating Urges: An Ecological Momentary Assessment Study.
The current study used ecological momentary assessment (EMA) to investigate whether appearance-based comments, social and performance-based evaluations affected levels of body dissatisfaction (BD) and urges to engage in disordered eating behaviors (DE) throughout daily life. A total of 620 participants completed a baseline questionnaire assessing sociodemographic variables. Participants then downloaded a mobile app which alerted them to complete short surveys assessing their levels of BD, DE urges, and experiences of receiving comments and evaluations six times per day for 7 days. Negative appearance-based comments predicted greater levels of state BD, while positive appearance comments predicted lower levels of state BD. Negative social and performance-based evaluations predicted an increase in state BD, while positive evaluations predicted a decrease in this outcome variable. No significant predictor was found for the DE urge outcomes. The present findings suggest that receiving negative and positive feedback in various domains of one's life may predict opposite outcomes for body image. However, these effects do not necessarily associate with urges to engage in DE in a nonclinical population.


Fetal alcohol spectrum disorders and the risk of crime.
Fetal alcohol spectrum disorders (FASD) are an important preventable global health concern. FASD is an umbrella term describing a range of mild to severe cognitive and behavioral problems among individuals prenatally exposed to alcohol. Alcohol causes FASD by interfering with molecular pathways during fetal development involving increased oxidative stress, disturbed organ development, and change of epigenetic gene expression control. Neuroimaging studies into FASD show several neuropathological abnormalities including abnormal brain structure, cortical development, white matter microstructure, and functional connectivity. Individuals with FASD experience a wide range of cognitive and behavioral challenges. Risks of violent behavior, criminality, and criminalization have been indicated by a limited number of epidemiological studies. The relationship between prenatal alcohol exposure and the increase of these risks remains unclear. This is further impeded by the complexity of an FASD diagnosis, the lack of a clear dose-response relationship of brain impact to alcohol use, and the lack of a clear FASD behavioral phenotype. Literature with respect to FASD and crime is still in its infancy. From the studies available, it is recommended to pay close attention to individuals with FASD and the relation with the criminal justice system and the risk for discrimination. There is a clear need for FASD-related stigma reduction programs within the correctional system. Further investigations into reliable biomarkers for diagnosis and treatment are needed.


Combination therapy for carbapenem-resistant Enterobacteriaceae: increasing evidence, unanswered questions, potential solutions.
Carbapenem-resistant Enterobacteriaceae (CRE) are associated with a high mortality rate and are an increasing problem worldwide. In this mini-review, we consider the growing number of observational studies in favour of combination therapy but highlight the absence of randomised control trials. We discuss the importance of data on minimum inhibitory concentrations (MICs), both for surveillance and for individual patient management. We examine the issues surrounding the use of carbapenems, polymyxins and tigecycline in the treatment of CRE. When and how should we be using carbapenems? Which polymyxin is best? Is tigecycline much maligned? Further studies are urgently needed to validate drug combinations, doses and ratios to maximise efficacy whilst reducing drug exposure and adverse effects.


Copper content of staple seeds and grains grown in Kanam local government area, Nigeria.
Copper is a trace mineral that plays an important role in human metabolism, largely because it allows many critical enzymes to function properly. Little is known of copper content of Nigerian foods. In this paper, copper contents of several typical Nigerian foods were determined. The samples were collected from Kanam of Plateau State in central region of Nigeria. The samples were wet-ashed according to the protocol of Hill et al. (Anal Chem 55:2340-2342, 1986).Concentrations of copper in the samples was determined using inductively coupled-mass spectrophotometry. There is, for each crop, a wide variation in copper content. The highest copper content was found in cowpea, Vigna unguiculata (16.95 μg/g of dry weight). Lowest copper content was found in white maize and in yellow maize (Zea mays), with values 1.23 μg/g and 1.38 μg/g of dry weights, respectively. Other foods, such as white sorghum, red sorghum (Sorghum bicolor), millet (Pennisetum glaucum), and groundnuts (Arachis hypogea) had copper contents varying from 2.22 to 11.81 μg/g. These values are well below the supplemental values of 50 mg/day that could interfere with zinc absorption. Thus, among the staple foodstuffs of the areas sampled, cowpea appears to be the richest source of dietary copper followed by groundnut while the two maize varieties are the poorest.


Autotaxin, an ectoenzyme that produces lysophosphatidic acid, promotes the entry of lymphocytes into secondary lymphoid organs.
The extracellular lysophospholipase D autotaxin (ATX) and its product, lysophosphatidic acid, have diverse functions in development and cancer, but little is known about their functions in the immune system. Here we found that ATX had high expression in the high endothelial venules of lymphoid organs and was secreted. Chemokine-activated lymphocytes expressed receptors with enhanced affinity for ATX, which provides a mechanism for targeting the secreted ATX to lymphocytes undergoing recruitment. Lysophosphatidic acid induced chemokinesis in T cells. Intravenous injection of enzymatically inactive ATX attenuated the homing of T cells to lymphoid tissues, probably through competition with endogenous ATX and exertion of a dominant negative effect. Our results support the idea of a new and general step in the homing cascade in which the ectoenzyme ATX facilitates the entry of lymphocytes into lymphoid organs.


Assessment of hepatotoxicity potential of drug candidate molecules including kinase inhibitors by hepatocyte imaging assay technology and bile flux imaging assay technology.
Kinases are members of a major protein family targeted for drug discovery and development. Given the ubiquitous nature of many kinases as well as the broad range of pathways controlled by these enzymes, early safety assessments of small molecule inhibitors of kinases are crucial in identifying new molecules with sufficient therapeutic window for clinical development. Failure or attrition of drug candidates in late-stage pipelines due to hepatotoxicity is a significant challenge in the drug development field. Herein we provide detailed methods for the hepatocyte imaging assay technology (HIAT) and the bile flux imaging assay technology (BIAT) to evaluate drug-induced liver injury (DILI) potentials for drug candidates. Optimized culturing methods for primary human hepatocytes, both freshly isolated and prequalified cryopreserved cells, are also presented. The applications of these high-content cellular imaging technologies in the evaluation of p38 and Her2 kinase inhibitors are highlighted to illustrate the usefulness of the research methodology in a compound screening as well as mechanistic investigative setting.


Inter-species differences for polychlorinated biphenyls and polybrominated diphenyl ethers in marine top predators from the Southern North Sea: Part 1. Accumulation patterns in harbour seals and harbour porpoises.
Harbour porpoises (Phocoena phocoena) and harbour seals (Phoca vitulina) are two representative top predator species of the North Sea ecosystem. The median values of sum of 21 polychlorinated biphenyl (PCB) congeners and sum of 10 polybrominated diphenyl ether (PBDE) congeners were 23.1 microg/g lipid weight (lw) and 0.33 microg/g lw in blubber of harbour seals (n=28) and 12.4 microg/g lw and 0.76 microg/g lw in blubber of harbour porpoises (n=35), respectively. For both species, the highest PCB concentrations were observed in adult males indicating bioaccumulation. On the contrary, the highest PBDE concentrations were measured in juveniles, likely due to better-developed metabolic capacities with age in adults. A higher contribution of lower chlorinated and non-persistent congeners, such as CB 52, CB 95, CB 101, and CB 149, together with higher contributions of other PBDE congeners than BDE 47, indicated that harbour porpoises are unable to metabolize these compounds. Harbour seals showed a higher ability to metabolize PCBs and PBDEs.


(2R)-8-Benzyl-2-[(S)-hy-droxy(phen-yl)meth-yl]-8-aza-bicyclo-[3.2.1]octan-3-one.
The crystal of the title compound, C(21)H(23)NO(2), was chosen from a conglomerate formed by a racemic mixture. An intra-molecular hydrogen bond is formed between hy-droxy group and heterocyclic N atom of the aza-bicyclo-[3.2.1]octan-3-one system. The crystal structure is stabilized by C-H⋯O inter-actions between aliphatic C-H groups and the carbonyl O atom. For the title chiral crystal, the highly redundant and accurate diffraction data set collected with low energy copper radiation gave a Flack parameter of 0.12 (18) for anomalous scattering effects originating from O atoms.


Retroperitoneal mature cystic teratoma arising from intra-abdominal undescended testis in an adult.
Teratomas are a type of germ-cell tumour which may contain embryological tissues from more than one germ-cell layer, usually occurring in the gonads. Primary retroperitoneal teratomas account for 1-11% of retroperitoneal neoplasms and are most commonly found in neonates and young adults. In the clinical literature, retroperitoneal mature cystic teratoma arising from an intra-abdominal undescended testis in adult males has been very rarely reported. We report a case of surgically proven retroperitoneal mature cystic teratoma arising from an intra-abdominal undescended testis in an adult male by multislice helical computed tomography and magnetic resonance imaging.


Small heterodimer partner interacting leucine zipper protein (SMILE) ameliorates autoimmune arthritis via AMPK signaling pathway and the regulation of B cell activation.
Rheumatoid arthritis (RA) is an autoimmune disease that causes joint swelling and inflammation and can involve the entire body. RA is characterized by the increase of pro-inflammatory cytokines such as interleukin (IL) and tumor necrosis factor, and the over-activation of T lymphocytes and B lymphocytes, which may lead to severe chronic inflammation of joints. However, despite numerous studies the pathogenesis and treatment of RA remain unresolved. This study investigated the use of small heterodimer partner-interacting leucine zipper protein (SMILE) overexpression to treat a mouse model of RA. SMILE is an insulin-inducible corepressor through adenosine monophosphate-activated kinase (AMPK) signaling pathway. The injection of a SMILE overexpression vector to mice with collagen induced-arthritis resulted in a milder clinical pathology and a reduced incidence of arthritis, less joint tissue damage, and lower levels of Th17 cells and plasma B cells in the spleen. Immunohistochemistry of the joint tissue showed that SMILE decreased B-cell activating factor (BAFF) receptor (BAFF-R), mTOR, and STAT3 expression but increased AMPK expression. In SMILE-overexpressing transgenic mice with collagen antibody-induced arthritis (CAIA), a decrease in the arthritis score and reductions in tissue damage, the number of B cells, and antibody production were observed. The treatment of immune cells in vitro with curcumin, a known SMILE-inducing agent, led to decreases in plasma B cells, germinal center B cells, IL-17-producing B cells, and BAFF-R-positive B cells. Taken together, our findings demonstrate the therapeutic potential of SMILE in RA, based on its inhibition of B cell activation mediated by the AMPK/mTOR and STAT3 signaling pathway and BAFF-R expression. Video abstract.


ATP and its metabolite adenosine act synergistically to mobilize intracellular calcium via the formation of inositol 1,4,5-trisphosphate in a smooth muscle cell line.
Interactions between ATP and adenosine on the formation of inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) and mobilization of intracellular calcium were investigated in the smooth muscle cell line DDT1 MF-2. Activation of adenosine A1 receptors with adenosine or cyclopentyladenosine (CPA) or of nucleotide receptors with ATP increased both Ins(1,4,5)P3 formation and intracellular calcium concentrations. The A1 receptor-induced Ins(1,4,5)P3 formation (EC50 10 nM) was antagonized by the A1 antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) and by pretreatment of the cells with pertussis toxin (PTX). ATP-stimulated Ins(1,4,5)P3 formation (EC50 21 microM) was attenuated, but still present, after PTX treatment. ATP and CPA had supraadditive effects on Ins(1,4,5)P3 accumulation and CPA increased ATP-induced Ins(1,4,5)P3 accumulation in a concentration-dependent manner with an EC50 of 3 nM, a concentration which per se had little or no effect on Ins(1,4,5)P3 accumulation. ATP (EC50 4 microM) and CPA (EC50 4 nM) both increased intracellular calcium levels. The effect of ATP was partially sensitive to PTX treatment, whereas the effect of CPA was blocked both by PTX and by DPCPX. Concentrations of ATP and CPA that by themselves were insufficient to raise intracellular calcium were able to do so when combined. The synergy between ATP and CPA on the mobilization of intracellular calcium was abolished after treatment of cells with PTX or when DPCPX was included in the experiment. Since ATP was metabolized by ecto-enzymes to ADP, AMP, and adenosine, we also examined whether adenosine formed from ATP could enhance the ATP effects on Ins(1,4,5)P3 accumulation. Indeed, the addition of the A1 receptor antagonist DPCPX or removal of endogenous adenosine by inclusion of adenosine deaminase in the experimental medium significantly attenuated the ATP response, and the two treatments did not have additive effects. The present study thus demonstrates that in a clonal cell line two types of receptors increase phospholipase C activity, but via different pathways; nucleotide receptors appeared to act via partially PTX-insensitive, and A1 receptors via PTX-sensitive G-proteins. ATP and CPA are not only able per se to induce formation of Ins(1,4,5)P3 and mobilize intracellular calcium, but they also act synergistically. Finally, it is demonstrated that endogenous adenosine, possibly formed from the rapid breakdown of ATP, can significantly enhance some ATP effects.


Changes in mitochondrial membrane potential and accumulation of reactive oxygen species precede ultrastructural changes during ovule abortion.
In many species, environmental stress reduces plant fertility. In Arabidopsis thaliana, a significant fraction of this reduction in plant fertility results from ovule abortion and embryo senescence. In this species, environmental conditions were identified that induced 94% of the developing ovules to either undergo stress-induced ovule abortion or embryo senescence (Sun et al. Plant Physiol 135:2358-2367, 2004). Following salt stress, physiological and anatomical changes were first detected in the female gametophyte of an aborting ovule. Two to four hours after a period of salt stress that induces most ovules to abort, the mitochondrial membrane potential dissipated. Subsequently, cells in the gametophyte accumulated reactive oxygen species, which are known to be molecules that promote programmed cell death (PCD). Because mitochondria often play an important role in PCD, these organelles were closely examined for changes in structure. Although the anatomy of mitochondria varied, reproducible changes in mitochondria structure were not observed. Nonetheless, other changes in ultrastructure were found. In some aborting gametophytes, concentric rings of endoplasmic reticulum were formed. In a fraction of the aborting ovules, cytoplasmic contents and organelles were invaginated into the vacuole. Even in cryofixed sections, many of these bodies appeared indistinct, which is consistent with the degradation of their contents.


Influence of initial fixation protocol on the appearance of primary cilia on the rabbit corneal endothelial cell apical surface as assessed by scanning electron microscopy.
The objective was to examine primary cilia at the apical surface of corneal endothelial cells after using different fixatives. Female albino rabbits (2 kg) were euthanised at 15:00 h and the corneas fixed immediately (usually with an isotonic 2% glutaraldehyde-cacodylate fixative) either after dissection, by application fixative at 4 degrees C, by immersion of the eyeball in fixative at room temperature (RT), or by application of an isotonic or a hypertonic (Karnovsky-type) fixative at RT. Images at 2000x were taken from the central corneal region, and number and length of primary cilia assessed. The length was the same regardless of method (overall average of 1.67+/-0.70 microm), but the incidence of primary cilia was hypertonic fixative (87% of cells) >cold drop fixation (71%), >whole globe immersion (68%) >dissect then fix methods (67%) >RT drop fixation (34%). The first four methods however yielded cells with unacceptable artefacts (especially distortion). More details should be provided of the primary fixation method used.


Perceived HIV stigma in AIDS caregiving dyads.
This study examines perceived HIV stigma in AIDS caregiving dyads in the United States, assessing the measurement of and correlates of personal stigma (among care-recipients living with HIV), courtesy stigma (among caregivers), and dyadic stigma. Survey data from 135 dyads in which the caregiver is a midlife or older mother or wife, and the care-recipient is her HIV-infected adult son or husband, are analyzed with individual-level and multilevel regression models. Results indicate that: (1) perceived stigma can be reliably measured among both persons living with HIV (PLH) and caregivers; (2) personal stigma can be distinguished from courtesy stigma; (3) perceived stigma is relatively low in this sample, and is higher among PLH than caregivers, higher among caregiving wives than mothers, and similar between PLH who are husbands and sons; (4) dyadic stigma is influenced by the caregiver's HIV status, the ethnic composition of the dyad, caregiving duration, and household income; (5) stigma discrepancy within dyads is a function of health discrepancy within dyads; and (6) differences in multivariate correlates of perceived stigma at the individual-level, in comparison to the dyad-level, suggest that dyadic stigma is a unique construct. A recognition that perceived stigma bears its own unique influence on the caregiving dyad is important for understanding how best to allocate resources aimed at alleviating stigma among individuals and families impacted by HIV.


Detection of neurocan in cerebrospinal fluid.
Cerebrospinal fluid (CFS) is the most easily accessible component of the human central nervous system and has been successfully used for the analysis of disease-associated molecular imbalances, particularly for extracellular matrix components. Alterations in the presence of the nervous system-associated chondroitin sulfate proteoglycan neurocan had been reported from active multiple sclerosis lesions. Neurocan could be detected as a component of human CFS after enrichment of proteoglycans by anion exchange chromatography from pooled liquor as well as individual 300 μL samples by Western blot. However, a general alteration in neurocan levels in CFS sample with high immunoglobulin content could not be demonstrated. To further reduce the sample size, the development of a PG capturing assay based on polybrene-coated 96-well plates was initiated. This approach could be an interesting alternative option for the analysis of PGs in biological fluid and tissue samples.


Diverse effects of estradiol-17 beta: concurrent suppression of appetite, blood pressure and vascular reactivity in conscious, unrestrained animals.
Evidence from epidemiological and clinical investigations have suggested a relationship between estrogen-containing oral contraceptives and hypertension. The present series of studies, however, documents the ability of estradiol-17 beta, a natural ovarian estrogen, to lower resting blood pressure and pressor responses to norepinephrine in conscious, unrestrained guinea pigs. Arterial measurements were made of resting blood pressure and heart rate, plus pressor responses to intravenous infusions of 1.56 micrograms norepinephrine. Injection of 30 micrograms estradiol-17 beta reduced resting pressures up to 12% and pressor responses up to 20% in the interval from 12 to 48 hours postinjection. The estradiol treatment also significantly and reversibly lowered food intake, water intake, and body weight. These effects could be induced by either 3 or 30 micrograms of estradiol benzoate for up to 4 days if estradiol treatment was continued. Parallel studies indicated that NE-induced contractions of the isolated aorta were markedly reduced by pretreatment with estradiol. These studies indicate that natural ovarian estrogens may reduce blood pressure by reducing the contractility of the arterial smooth muscle.


Factors associated with congenital syphilis: A retrospective study in Jiangxi Province, China.
There are a lack of studies about factors influencing congenital syphilis (CS) in economically underdeveloped areas, such as Jiangxi Province, China. A retrospective study was conducted based on the information system of prevention of mother-to-child transmission of syphilis management in Jiangxi Province, China. Pregnant women with syphilis infection who delivered ≥28 gestational weeks and registered in this system from 1 January 2013 to 2030 June 2018 were enrolled. Maternal characteristics and treatment regimens associated with CS were evaluated using multivariable regression analysis. 1196 syphilis infected mothers and their 1207 infants were included in the analyses, and 116 infants were diagnosed with CS, providing an overall incidence of 9.61% (116/1207). Multivariable logistic regression analysis showed that increasing maternal age was barely associated with the risk of CS (adjusted odds ratio (aOR) = 0.97, 95% CI, 0.93-1.00, p = .047). Women with a high nontreponemal serum test titer (≥1:8) had a 126% increased risk of delivering an infant with CS than those with a low titer (<1:8) (aOR = 2.26, 95% CI, 1.51-3.39, p < .001). The risk for CS decreased significantly in infants born to mothers receiving adequate treatment than those receiving no treatment (aOR = 0.36, 95% CI, 0.21-0.61, p < .001). Adequate treatment is critical for the prevention of CS. Further strategies focusing on early diagnosis and adequate treatment among syphilis infected pregnant women, particularly among those with younger age and high nontreponemal titer, should be strengthened to prevent CS.


Nanoparticle designs for delivery of nucleic acid therapeutics as brain cancer therapies.
Glioblastoma (GBM) is an aggressive central nervous system cancer with a dismal prognosis. The standard of care involves surgical resection followed by radiotherapy and chemotherapy, but five-year survival is only 5.6% despite these measures. Novel therapeutic approaches, such as immunotherapies, targeted therapies, and gene therapies, have been explored to attempt to extend survival for patients. Nanoparticles have been receiving increasing attention as promising vehicles for non-viral nucleic acid delivery in the context of GBM, though delivery is often limited by low blood-brain barrier permeability, particle instability, and low trafficking to target brain structures and cells. In this review, nanoparticle design considerations and new advances to overcome nucleic acid delivery challenges to treat brain cancer are summarized and discussed.


Quantitative assessment of emphysema and bronchial wall thickness in patients with stable chronic obstructive pulmonary disease: comparison between the eosinophilic and non-eosinophilic phenotypes.
To perform a quantitative assessment of bronchial wall thickening and the emphysema score in patients with stable chronic obstructive pulmonary disease (COPD), comparing the eosinophilic and non-eosinophilic COPD phenotypes. This was a retrospective observational study of patients with COPD followed between August 2018 and July 2019. The patients were divided into two groups by the eosinophil count in peripheral blood: eosinophilic (≥ 300 cells/µL); and non-eosinophilic (< 300 cells/µL). Quantitative, automated assessments of emphysema and bronchial wall thickness were performed by evaluating computed tomography scans of the chest. We evaluated the records of 110 patients diagnosed with COPD: 28 (25.5%) in the eosinophilic group; and 82 (74.5%) in the non-eosinophilic group. The demographic, clinical, functional, and therapeutic variables were comparable between the two groups. There were no significant differences between the two groups in terms of the emphysema score or bronchial wall thickness (p > 0.05 for both). Patients with eosinophilic COPD do not appear to have lower emphysema scores or greater bronchial wall thickening than do those with non-eosinophilic phenotypes of the disease.


Clinical outcomes, quality of life, and costs associated with implantation of vagus nerve stimulation therapy in pediatric patients with drug-resistant epilepsy.
VNS (Vagus Nerve Stimulation Therapy) is approved in the USA to treat refractory epilepsy as adjunctive to antiepileptic drugs (AEDs) in patients ≥12 years with complex partial seizures. To evaluate clinical outcomes, quality-adjusted life years (QALY), and costs associated with VNS in pediatric patients with drug-resistant epilepsy in a real-world setting. A retrospective analysis was conducted using Medicaid data (USA). Patients had ≥1 neurologist visits with epilepsy diagnosis (ICD-9 345.xx, 780.3x), ≥1 procedure claims for VNS implantation, ≥1 AEDs, ≥6-months of Pre- and Post-VNS continuous enrollment. Pre-VNS period was 6-months and Post-VNS period extended from implantation until device removal, death, Medicaid disenrollment, or study end (up to 3 years). Incidence rate ratios (IRR) and costs ($2010) were estimated. QALYs were estimated using number of seizure-related events. For patients 1-11 years old (N = 238), hospitalizations and emergency room visits were reduced Post-VNS vs. Pre-VNS (adjusted IRR = 0.73 [95% CI: 0.61-0.88] and 0.74 [95% CI: 0.65-0.83], respectively). Average total healthcare costs were lower Post-VNS vs. Pre-VNS ($18,437 vs. $18,839 quarterly [adjusted p = 0.052]). For patients 12-17 years old (N = 207), hospitalizations and status epilepticus events were reduced Post-VNS vs. Pre-VNS (adjusted IRR = 0.43 [95% CI: 0.34-0.54] and 0.25 [95% CI: 0.16-0.39], respectively). Average total healthcare costs were lower Post-VNS vs. Pre-VNS period ($14,546 vs. $19,695 quarterly [adjusted p = 0.002]). Lifetime QALY gain after VNS was 5.96 (patients 1-11 years) and 4.82 years (patients 12-17 years). VNS in pediatric patients is associated with decreased resource use and epilepsy-related events, cost savings, and QALY gain.


Topical therapies for diabetic foot ulcers: standard treatments.
This paper is the first in a two-part series reviewing the evidence on standard and advanced treatments for diabetic foot ulcers. Both clinical studies and economic models are evaluated. The first part concentrates on standard topical therapies.