Intradermal substance P as a challenge agent in healthy individuals.
Pharmacological challenge models are deployed to evaluate drug effects during clinical development. Intradermal injection of Substance P (SP) neuropeptide, a potential challenge agent for investigating local mediators, is associated with wheal and flare response mediated by the MRGPRX2 receptor. Although dose-dependent data on SP effects exist, full characterization and information on potential carryover effect after repeated challenge are lacking. This open-label, two-part, prospective enabling study of SP intradermal challenge in healthy participants aimed to understand and distinguish between wheal and flare responses following various SP doses. Part 1 included one challenge visit to determine optimum SP dose range for evaluation in part 2, which determined variability in 20 participants and used intradermal microdialysis (IDM) for SP-challenged skin sampling. At 5, 15, 50, and 150 pmol doses, respectively, posterior median area under the curve (AUC; AUC0-2h ) was 4090.4, 5881.2, 8846.8, and 9212.8 mm2 /min, for wheal response, and 12020.9, 38154.3, 65470.6, and 67404.4 mm2 /min for flare response (SP-challenge visit 2). When the challenge was repeated ~2 weeks later, no carryover effect was observed. IDM histamine levels were relatively low, resulting in low confidence in the data to define temporal characteristics for histamine release following SP challenge. No safety concerns were identified using SP. Wheal and flare responses following intradermal SP challenge were dose-dependent and different. The results indicate that this challenge model is fit-for-purpose in future first-in-human studies and further assessment of novel drugs targeting dermal inflammatory disease responses, such as chronic spontaneous urticaria, chronic inducible urticaria, and pseudo-allergic reactions.


Xylan degradation by the thermophile Clostridium stercorarium: cloning and expression of xylanase, beta-D-xylosidase, and alpha-L-arabinofuranosidase genes in Escherichia coli.
Seven genes related to arabinoxylan degradation were isolated from a genomic library of the thermophilic bacterium Clostridium stercorarium. The cloned genes include a xylanase gene (xynA), two beta-D-xylosidase genes (bx1A and bx1B), two alpha-L-arabinofuranosidase genes (arfA and arfB), and two genes (celW and celX) encoding enzymes termed celloxylanases, which hydrolyze both xylans and beta-D-cellobiosides. The genes xynA, celX, and bxlB were found to encode the major xylanolytic enzyme activities induced by growth of C. stercorarium on xylan.


Glomerular proteomic profiling reveals early differences between preexisting and de novo type 2 diabetes in human renal allografts.
Diabetes mellitus (DM), either preexisting or developing after transplantation, remains a crucial clinical problem in kidney transplantation. To obtain insights into the molecular mechanisms underlying PTDM development and early glomerular damage before the development of histologically visible diabetic kidney disease, we comparatively analysed the proteome of histologically normal glomeruli from patients with PTDM and normoglycaemic (NG) transplant recipients. Moreover, to assess specificities inherent in PTDM, we also comparatively evaluated glomerular proteomes from transplant recipients with preexisting type 2 DM (T2DM). Protocol biopsies were obtained from adult NG, PTDM and T2DM patients one year after kidney transplantation. Biopsies were formalin-fixed and embedded in paraffin, and glomerular cross-sections were microdissected. A total of 4 NG, 7 PTDM and 6 T2DM kidney biopsies were used for the analysis. The proteome was determined by liquid chromatography-tandem mass spectrometry. Relative differences in protein abundance and significantly dysregulated pathways were analysed. Proteins involved in cell adhesion, immune response, leukocyte transendothelial filtration, and cell localization and organization were less abundant in glomeruli from PTDM patients than in those from NG patients, and proteins associated with supramolecular fibre organization and protein-containing complex binding were more abundant in PTDM patients. Overall, proteins related to adherens and tight junctions and those related to the immune system, including leukocyte transendothelial migration, were more abundant in NG patients than in transplanted patients with DM, irrespective of the timing of its development. However, proteins included in cell‒cell junctions and adhesion, insulin resistance, and vesicle-mediated transport were all less abundant in PTDM patients than in T2DM patients. The glomerular proteome profile differentiates PTDM from NG and T2DM, suggesting specific pathogenetic mechanisms. Further studies are warranted to validate these results, potentially leading to an improved understanding of PTDM kidney transplant pathophysiology and to the identification of novel biomarkers.


Tremor Mimicking Ventricular Tachycardia on Telemetry.
A 79-year-old man with coronary artery disease and tremor-predominant Parkinson's disease underwent a gross total resection of a high-grade glioma and exhibited what appeared to be polymorphic ventricular tachycardia lasting 15-20 seconds on postoperative day 1. Further evaluation revealed that the patient did not have ventricular tachycardia, and that his abnormal telemetry signals were instead an artifact of his Parkinson's tremor. This case underscores the importance of considering tremor artifact when evaluating abnormal telemetry and electrocardiogram signals in patients with tremor, and highlights some features that can distinguish tremor artifact from a true arrhythmia.


Mitochondria-rich (chloride) cells in the gill epithelia from four species of stenohaline fresh water teleosts.
The mitochondria-rich (chloride) cells have been found to be present in the gill epithelia of four species of stenohaline fresh water teleosts. The cytoplasm of these chloride cells contains an extensive network of cytoplasmic tubules which communicate with intercellular spaces bordering the lateral and basal cell surfaces. Numerous vesicles with fairly electron-dense interiors are also present in the apical cytoplasm of chloride cells. The apical surface of a chloride cell forms an apical pit, but the lumen of the pit does not appear to be in continuity with the interior of the apical vesicles and tubules inside the cell. When Carassius auratus were kept in 100, 200, 300, and 400 mOsm-diluted sea water for a month, no appreciable changes occurred in the number and fine structure of the chloride cells, except for a dilation of the apical vesicles and a slight decrease in diameter of the cytoplasmic tubules in these cells in the fishes kept in 300 and 400 mOsm. These results suggest that chloride cells may be a rather common occurrence in the gill epithelia of stenohaline fresh water teleosts, and may function in ion transport in these fishes in fresh water environments.


Computer-aided diagnosis for the detection and classification of lung cancers on chest radiographs ROC analysis of radiologists' performance.
The aim of the study is to investigate the effect of a computer-aided diagnostic (CAD) scheme on radiologist performance in the detection of lung cancers on chest radiographs. We combined two independent CAD schemes for the detection and classification of lung nodules into one new CAD scheme by use of a database of 150 chest images, including 108 cases with solitary pulmonary nodules and 42 cases without nodules. For the observer study, we selected 48 chest images, including 24 lung cancers, 12 benign nodules, and 12 cases without nodules, from the database to investigate radiologist performance in the detection of lung cancers. Nine radiologists participated in a receiver operating characteristic (ROC) study in which cases were interpreted first without and then with computer output, which indicated locations of possible lung nodules, together with a five-color scale illustrating the computer-estimated likelihood of malignancy of the detected nodules. Performance of the CAD scheme indicated that sensitivity in detecting lung nodules was 80.6%, with 1.2 false-positive results per image, and sensitivity and specificity for classification of nodules by use of the same database for training and testing the CAD scheme were 87.7% and 66.7%, respectively. Average area under the ROC curve value for detection of lung cancers improved significantly (P = .008) from without (0.724) to with CAD (0.778). This type of CAD scheme, which includes two functions, namely detection and classification, can improve radiologist accuracy in the diagnosis of lung cancer.


[A differential approach to applying the terrain cure to adolescents with arterial hypertension].
The "severity index" obtained by specially devised algorithm and the type of central circulation served as the basis for adjustment of terrain exercise regimens in the treatment of hypertensive adolescents. The distance of the walk and the route topography were chosen differentially according to cardiac performance of each patient reflected in the above severity index of the disease.


Bone trace element pattern in an 18th century population sample of Tenerife (Canary Islands): comparison with a prehistoric one.
We have determined bone strontium (Sr), barium (Ba), calcium (Ca), and zinc (Zn) content in 24 samples belonging to adult individuals who died toward the end of the 18th century and were interred in a church's floor on the island of Tenerife, comparing the results with those obtained in 14 prehistoric samples of the same island and also with those of 7 modern controls. No differences were observed between the two ancient groups, which showed higher bone strontium and barium than the modern sample, and a slightly lower Ba/Sr ratio, thus pointing to consumption of marine sources.


Canola and hydrogenated soybean oils accelerate ectopic bone formation induced by implantation of bone morphogenetic protein in mice.
Canola oil (Can) and hydrogenated soybean oil (H2-Soy) are commonly used edible oils. However, in contrast to soybean oil (Soy), they shorten the survival of stroke-prone spontaneously hypertensive (SHRSP) rats. It has been proposed that the adverse effects of these oils on the kidney and testis are caused at least in part by dihydro-vitamin K (VK) 1 in H2-Soy and unidentified component(s) in Can. Increased intake of dihydro-VK1 is associated with decreased tissue VK2 levels and bone mineral density in rats and humans, respectively. The aim of the present study was to determine the effects of these oils on bone morphogenetic protein (BMP)-induced ectopic bone formation, which is promoted by VK2 deficiency, in relation to the role of VK in the γ-carboxylation of osteocalcin and matrix Gla protein. A crude extract of BMPs was implanted into a gap in the fascia of the femoral muscle in 5-week-old mice maintained on a Soy, Can, or H2-Soy diet. Newly formed bone volume, assessed by three-dimensional X-ray micro-computed tomography and three-dimensional reconstruction imaging for bone, was 4-fold greater in the Can and H2-Soy groups than in the Soy group. The plasma carboxylated osteocalcin (Gla-OC) and total OC (Gla-OC plus undercarboxylated osteocalcin [Glu-OC]) levels were significantly lower in the Can group than in the Soy group (p < 0.05). However, these levels did not significantly differ between the H2-Soy and Soy groups. The plasma Gla-OC/Glu-OC ratio in the Can and H2-Soy groups was significantly lower (in Can; p = 0.044) or was almost significantly lower (in H2-Soy; p = 0.053) than that in the Soy group. In conclusion, Can and H2-Soy accelerated BMP-induced bone formation in mice to a greater extent than Soy. Further research is required to evaluate whether the difference in accelerated ectopic bone formation is associated with altered levels of VK2 and VK-dependent protein(s) among the three dietary groups.


Higher levels of social and material deprivation are associated with higher rates of influenza-like illness-related emergency department visits: Edmonton, Alberta, 2004-2014.
Neighbourhood-level deprivation is associated with hospitalization related to respiratory infections; however, hospitalizations exclude many who seek care with less severe respiratory illnesses. Another major seasonal contributor to respiratory illness-associated healthcare burdens are influenza-like illness (ILI)-related emergency department (ED) visits. We investigated associations between area-level social and material deprivation and ILI-related ED use. This is a retrospective ecological study. We linked ILI-related ED visit data (2004-2014) for Edmonton, Alberta to a Canadian area-level material and social deprivation index, categorizing deprivation into quintiles. Using a multivariable Poisson model with log population as the offset, we modelled the relationship between visit rates and material and social deprivation adjusting for week and season, age, sex and the interaction between age and sex. We included 67,585 ILI-related ED visits, representing 1075.5 (95% confidence interval (CI) = 1067.4-1083.6) visits per 100,000 person-years. ILI-related visit rates increased as each of material and social deprivation increased; increases were slightly greater for material deprivation. Comparing the most deprived quintile to the least deprived quintile: for material deprivation, ILI-related ED visit rates were two times higher (rate ratio (RR) = 2.00, 95% CI = 1.96-2.05); and, for social deprivation, one-and-a-half times higher (RR = 1.47, 95% CI = 1.44-1.51). Higher area-level material and social deprivation were associated with higher ILI-related ED visit rates. These findings can be used to identify areas that may need additional public health and healthcare resources and to improve targeting of prevention strategies. Understanding differentials in healthcare use such as this may be especially relevant to ensuring equity of outcomes for pandemic preparedness planning.


Biological and environmental factors may affect children's executive function through motor and sensorimotor development: Preterm birth and cerebral palsy.
Disruptive biological and environmental factors may undermine the development of children's motor and sensorimotor skills. Since the development of cognitive skills, including executive function, is grounded in early motor and sensorimotor experiences, early delays or impairments in motor and sensorimotor processing often trigger dynamic developmental cascades that lead to suboptimal executive function outcomes. The purpose of this perspective paper is to link early differences in motor/sensorimotor processing to the development of executive function in children born preterm or with cerebral palsy. Uncovering such links in clinical populations would improve our understanding of developmental pathways and key motor and sensorimotor skills that are antecedent and foundational for the development of executive function. This knowledge will allow the refinement of early interventions targeting motor and sensorimotor skills with the goal of proactively improving executive function outcomes in at-risk populations.


The different central nervous system efficacy among gefitinib, erlotinib and afatinib in patients with epidermal growth factor receptor mutation-positive non-small cell lung cancer.
Brain metastasis is common in non-small cell lung cancer (NSCLC) and has an even higher incidence in epidermal growth factor receptor (EGFR)-mutant cancers. Although EGFR tyrosine kinase inhibitors (TKIs) are effective against brain metastases, it is unknown which first- or second-generation EGFR TKI is most effective. Patients treated with first-line gefitinib, erlotinib, or afatinib for advanced EGFR-mutant NSCLC were included. The efficacy against brain metastasis was evaluated by comparing the response rates of measurable and non-irradiated brain metastases, central nervous system progression-free survival (CNS-PFS), and the cumulative incidence of CNS failure. Among the 559 patients who received EGFR-TKIs (gefitinib, n=299; erlotinib, n=93; afatinib, n=167), 198 had initial brain metastasis before starting EGFR-TKIs. The CNS response rates of gefitinib, erlotinib, and afatinib were 64.7%, 68.2%, and 72.9%, respectively (P=0.78). In the overall study population, irrespective of initial CNS metastasis, the median CNS-PFS was 17.3 months for gefitinib, 12.4 months for erlotinib, and 23.3 months for afatinib (P<0.001). In multivariate analysis for CNS-PFS, the hazard ratio (HR) of afatinib was 0.63 (95% CI, 0.47-0.83) compared with gefitinib or erlotinib. In the competing risk analysis for cumulative incidence of CNS failure, afatinib showed a lower cumulative incidence of CNS failure compared with gefitinib or erlotinib after adjusting for both EGFR mutation type and preexisting CNS metastases (HR 0.51, 95% CI, 0.34-0.75, P=0.0007). Through there are some limitation as a retrospective study, afatinib showed similar CNS response rates, superior CNS-PFS and cumulative incidence of CNS failure, compared with gefitinib or erlotinib.


Solubilization and reconstitution of the sarcolemmal Na+-Ca2+ exchange system of vascular smooth muscle.
The Na+-Ca2+ exchange system of the sarcolemma of rat mesenteric artery was solubilized and reconstituted in soybean phospholipid vesicles. In the reconstituted system, the exchange process showed about 4-fold higher specific activity compared to that of native vesicles. The inhibitory effect of monensin and the stimulatory effect of valinomycin in the presence of K+ on Na+ gradient-dependent Ca2+ uptake were preserved and were pronounced in the reconstituted system. The stimulation by valinomycin indicates that the exchange process is electrogenic. Thus, the stoichiometry, the characteristics and the mechanism of action which were difficult to study in the native vesicles can now be determined conveniently using the reconstituted system. Also, solubilization and reconstitution of the exchange system confirms its existence in vascular smooth muscle.


Impact of the COVID-19 pandemic in patients with a previous history of premature myocardial infarction.
The coronavirus-disease-2019 (COVID-19) pandemic has led to the restructuring of health-services to prioritize the treatment of COVID-19. The severe restrictions on daily life affected the management of chronic diseases. Patients with a previous history of premature myocardial infarction (MI) are a vulnerable group requiring frequent and continued medical attention both in the pandemic and non-pandemic era. The present study was conducted to provide insight into the impact of COVID-19 outbreak on heart-healthy lifestyle and management of patients with a history of premature MI. This cross-sectional study included 170 consecutive patients with a history of premature MI who were already in regular follow-up in a tertiary out-patient prevention clinic before the pandemic. Inclusion criteria included age ≥18 years and being on regular follow up with the diagnosis of premature MI (documented MI before the age of 55 years) at least for one year. All patients were contacted by phone-call and replied to a 23-item questionnaire measuring the impact of the pandemic on the management, healthy lifestyle habits, and anxiety level. One patient died due to COVID-19 infection; therefore the analyses were conducted in 169 patients (age: 47.67 ± 11.84 years, 21.3% women). The median age at first MI was 39 (IQR 10) years and the median time elapsed since the first MI was 7 years (IQR 10). The study population was highly compliant with the follow-up visits (78.1%) and pharmacological therapy (97%) before the pandemic according to the medical files. The majority (82.2%) of the patients were aware that having a history of premature MI would increase the risk and harm of COVID-19. Anxiety level increased in 62.7% of the study patients. Overall, 65.7% of the patient group reported a disruption at least in ≥1 component(s) of healthy life-behaviors (non-compliance with the heart-healthy diet, an increase in alcohol intake, an increase in smoking, and/or reduced physical activity) since the emergence of the outbreak. The anxiety level (p = 0.001) and the prevalence of appetite change (p < 0.0001) and weight gain (p < 0.0001) was lower in the lifestyle compliant group than the non-compliant group. Avoidance of seeking medical care was reported in 33.7% of the patients. Statin use was 99.4% before the pandemic and decreased to 89.9% (p < 0.0001) despite the fact that medications were reimbursed and widely available. The COVID-19 pandemic significantly affected the heart-healthy lifestyle and anxiety levels of patients with a history of premature MI who were already in regular follow-up in a tertiary prevention clinic and led to significant avoidance of medical care. More rigorous follow-up, education, and reassurance of these patients with telemedicine are necessary for the prevention of further increase in their risk.


[Basic substituted thieno(3,4-d)pyrimidine carboxylic acids: synthesis via ANRORC-reaction starting from 7-cyanthieno(3,2-d)oxazines].
The synthesis of 7-cyan-thieno[3,2-d]-1,3-oxazine 7 is reported. Reaction of 7 with primary amines gives the thieno[3,4-d]pyrimidincarboxylic acids 14 in a unusual ring transformation. Several synthesized compounds were tested as antiallergic agents.


Putting AlphaFold models to work with phenix.process_predicted_model and ISOLDE.
AlphaFold has recently become an important tool in providing models for experimental structure determination by X-ray crystallography and cryo-EM. Large parts of the predicted models typically approach the accuracy of experimentally determined structures, although there are frequently local errors and errors in the relative orientations of domains. Importantly, residues in the model of a protein predicted by AlphaFold are tagged with a predicted local distance difference test score, informing users about which regions of the structure are predicted with less confidence. AlphaFold also produces a predicted aligned error matrix indicating its confidence in the relative positions of each pair of residues in the predicted model. The phenix.process_predicted_model tool downweights or removes low-confidence residues and can break a model into confidently predicted domains in preparation for molecular replacement or cryo-EM docking. These confidence metrics are further used in ISOLDE to weight torsion and atom-atom distance restraints, allowing the complete AlphaFold model to be interactively rearranged to match the docked fragments and reducing the need for the rebuilding of connecting regions.


2-thio-6-azauridine inhibits Vpu mediated BST-2 degradation.
BST-2 is an interferon-induced host restriction factor that inhibits the release of diverse mammalian enveloped viruses from infected cells by physically trapping the newly formed virions onto the host cell surface. Human Immunodeficiency Virus-1 (HIV-1) encodes an accessory protein Vpu that antagonizes BST-2 by down-regulating BST-2 from the cell surface. Using a cell-based ELISA screening system, we have discovered a lead compound, 2-thio-6-azauridine, that restores cell surface BST-2 level in the presence of Vpu. This compound has no effect on the expression of BST-2 and Vpu, but inhibits Vpu-mediated BST-2 down-regulation and exerts no effect on Vpu-induced down-regulation of CD4 or KSHV K5 protein induced BST-2 down-regulation. 2-thio-6-azauridine suppresses HIV-1 production in a BST-2-dependent manner. Further results indicate that 2-thio-6-azauridine does not interrupt the interaction of BST-2 with Vpu and β-TrCP2, but decreases BST-2 ubiquitination. Our study demonstrates the feasibility of using small molecules to target Vpu function and sensitize wild type HIV-1 to BST-2-mediated host restriction.


From gene networks to brain networks.
The brain's structural organization is so complex that 2,500 years of analysis leaves pervasive uncertainty about (i) the identity of its basic parts (regions with their neuronal cell types and pathways interconnecting them), (ii) nomenclature, (iii) systematic classification of the parts with respect to topographic relationships and functional systems and (iv) the reliability of the connectional data itself. Here we present a prototype knowledge management system (http://brancusi.usc.edu/bkms/) for analyzing the architecture of brain networks in a systematic, interactive and extendable way. It supports alternative interpretations and models, is based on fully referenced and annotated data and can interact with genomic and functional knowledge management systems through web services protocols.


Tissue reaction to absorbable endoloop, nonabsorbable titanium staples, and polymer Hem-o-lok clip after laparoscopic appendectomy.
The standard technique for securing the base of the appendix during laparoscopic appendectomy is by absorbable endoloop ligature, although clinical reports favor the use of the stapler. Nonabsorbable Hem-o-lok clips have been shown to be an alternative technique to this. However, it is currently not clear whether nonabsorbable clips have any effects on the intestine or promote infection in the surgical area. Sixty Wistar albino rats were randomized into 3 treatment groups: group I (n=20) the base of the appendix was secured by endoloop 2-0 ligature; group II (n=20) dissection of the appendix was performed by a 45-mm thick stapler; and group III (n=20) the base of the appendix was secured by a Hem-o-lok plastic clip. The animals were sacrificed on the 14th and 28th days after surgery. The secured stump was used for histopathological examination. There were no significant differences in histopathological changes observed on the 14th postoperative day between the groups. On the 28th postoperative day, it was proved that mild and moderate inflammation is more frequent in the endoloop and Hem-o-lok groups than in the stapler group. Reaction to a foreign body is more frequent in the endoloop than in stapler and Hem-o-lok groups. The mildest postoperative inflammatory changes were seen in the stapler group, followed by the Hem-o-lok group. However, because of the price of the plastic clip and the simplicity of its application, its use is still favored during laparoscopic appendectomy.


Roentgenologic diagnosis of pancreatic disease. By Josef Rösch, 1967.
On the basis of his own experience and of literary sources, the author reports the present situation of the roentgenologic diagnosis of the pancreas. He evaluates the various examination methods and gives a general description of the typical characteristics of the diseases of the pancreas. In conclusion, he suggests ways for further improvements.


Regional differences in resting extracellular potassium levels of rat brain.
The extracellular potassium concentration in the cerebral cortex of the mammalian brain has been repeatedly reported to be approximately 3.0 mM. We have made detailed measurements with potassium-selective microelectrodes and have found significantly lower extracellular potassium concentrations in unstimulated rat brain caudate and thalamus (1.9-2.5 mM) when compared to cortex and cerebral spinal fluid (3.0-3.5 mM). These regional differences may be caused by variations in spontaneous activity of neurons, regional permeability differences in endothelial cells of brain capillaries to potassium, or caused by variations in uptake by glia.


Relevance of mangled extremity severity score to compartment syndromes.
This study investigated the relevance of a mangled extremity severity score (MESS) to the evaluation of intracompartmental pressure in injured patients. We measured intracompartmental pressures with a modification of the Whitesides method and compared these values with MESS in 27 patients after trauma. For 5 of them fasciotomy was necessary to relieve high intracompartmental pressures. In our series MESS seemed to correlate with intracompartmental pressure values (r = 0.733, P < 0.05). We conclude that the MESS system provides objective criteria in determining the risk of a compartment syndrome.


ARF-GEF cytohesin-2/ARNO regulates R-Ras and α5-integrin recycling through an EHD1-positive compartment.
When expressed in epithelial cells, cytohesin-2/ARNO, a guanine nucleotide exchange factor (GEF) for ARF small GTPases, causes a robust migration response. Recent evidence suggests that cytohesin-2/ARNO acts downstream of small the GTPase R-Ras to promote spreading and migration. We hypothesized that cytohesin-2/ARNO could transmit R-Ras signals by regulating the recycling of R-Ras through ARF activation. We found that Eps15-homology domain 1 (EHD1), a protein that associates with the endocytic recycling compartment (ERC), colocalizes with active R-Ras in transiently expressed HeLa cells. In addition, we show that EHD1-positive recycling endosomes are a novel compartment for cytohesin-2/ARNO. Knockdown or expression of GEF-inactive (E156K) cytohesin-2/ARNO causes R-Ras to accumulate on recycling endosomes containing EHD1 and inhibits cell spreading. E156K-ARNO also causes a reduction in focal adhesion size and number. Finally, we demonstrate that R-Ras/ARNO signaling is required for recycling of α5-integrin and R-Ras to the plasma membrane. These data establish a role for cytohesin-2/ARNO as a regulator of R-Ras and integrin recycling and suggest that ARF-regulated trafficking of R-Ras is required for R-Ras-dependent effects on spreading and adhesion formation.


Conditional reasoning, causality, and the structure of semantic memory: strength of association as a predictive factor for content effects.
Available evidence indicates that responses to conditional inferences using concrete causal premises is affected by the relative number of available alternate causes (Cummins, D.D., 1995. Memory and Cognition 23 (5), 646-658). We propose that another important factor that may influence the kinds of inferences made to causal conditionals is the relative strength of association between such causes and the consequent term. We present a study with adult participants that examines the effect of strength of association on performance on a conditional reasoning task using causal premises for which there exist one highly associated potential cause for the given consequent term. We predicted that adults would produce a greater proportion of biconditional responses to invalid forms with strongly associated premises than weakly associated ones, while valid forms would not be affected by strength of association. The results are consistent with this hypothesis.


The role of transperineal ultrasonography for perinatal management: A review.
Transperineal ultrasound (TPUS) has become an increasingly popular tool in obstetrics due to its objective, non-invasive, and real-time imaging capabilities. This review aims to describe the basic approaches, current utilization, and potential future applications of TPUS. A comprehensive literature review on TPUS was conducted. In addition, discussions at academic meetings and congress focused on TPUS were also considered. TPUS was initially used in prostate biopsies and is currently applied to evaluating fetal head descent in labor, with the angle of progression being the most widely used parameter. It is more tolerated than conventional invasive or expensive methods, such as digital vaginal examinations or MRIs. Additionally, TPUS can assess the internal rotation of the fetal head in the birth canal. Compared to other imaging modalities like MRI and CT scans, TPUS is easier to perform and more cost-effective. It also provides real-time imaging, allowing for quick and accurate assessments. It also help clinicians make critical decisions regarding the mode of delivery and identify patients at high risk for fecal incontinence postpartum. With its many benefits, TPUS has the potential to become a standard tool in urogynecology and obstetrics. Transperineal ultrasound is a non-invasive imaging modality that is well-tolerated and easy to understand for patients and their family and help medical staff support the patients. Transperineal ultrasound can be applied in real-time monitoring of labor progress, helping predict the possibility of vaginal delivery during labor, and further research in this area is warranted.


Alphaviruses in cancer immunotherapy.
Alphaviruses have frequently been engineered for cancer therapy, cancer immunotherapy, and cancer vaccine development. As members of self-replicating RNA viruses, alphaviruses provide high levels of transgene expression through efficient self-amplifying of their RNA genome in host cells. Alphavirus vectors can be used as recombinant viral particles or oncolytic viruses. Alternatively, either naked or nanoparticle-encapsulated RNA and DNA replicons can be utilized. In the context of cancer prevention and treatment, antitumor, cytotoxic and suicide genes have been expressed from alphavirus vectors to provide tumor regression and tumor eradication. Moreover, immunostimulatory genes such as cytokines and chemokines have been used for cancer immunotherapy approaches. Expression of tumor antigens has been applied for cancer vaccine development. Alphavirus vectors has demonstrated tumor regression and even cure in various preclinical animal models. Immunization has elicited strong immune responses and showed protection against challenges with tumor cells in animal models. Several clinical trials have confirmed good safety and tolerability of alphaviruses in cancer patients although therapeutic efficacy will still require optimization.


Infantile autism and computerized tomography brain-scan findings: specific versus nonspecific abnormalities.
The hypothesis that specific computerized tomography brain-scan findings are associated with infantile autism was tested in 45 cases and 19 controls. The autistic group was subdivided into serious and less-serious language-impaired subgroups. The analysis of Euclidean Distances, a type of cluster analysis, showed that neuroradiological parameters of cases and controls, including ventricular sizes, were on the whole significantly different, but no statistically significant difference appeared between the two autistic subgroups. But the analysis of variance of each neuroradiological parameter did not show any significant difference between autistics and controls. It was concluded that autism is nonspecifically associated with brain-scan abnormalities, and that other nonorganic, as well as organic, factors should be taken into account.


Measurement by intracoronary ultrasound of in vivo arterial distensibility within atherosclerotic lesions.
Arterial distensibility is diminished by atherosclerosis. This process has not been well studied in the coronary arteries. The purpose of this study was to assess changes in coronary arterial distensibility in 4 groups of patients. Group I (n = 20) consisted of patients with normal vessels, group II (n = 40) with diseased undilated vessels, group III (n = 15) after successful percutaneous transluminal coronary angioplasty (PTCA), and Group IV (n = 20) after successful directional coronary atherectomy (DCA). Intracoronary ultrasound imaging was used to assess distensibility, plaque morphology and atherosclerotic burden (expressed as the percentage of total vessel cross-sectional area occupied by plaque: percent plaque area). Distensibility was defined as percent change in lumen area in a cardiac cycle. Group I (normal vessels) had a distensibility = 14 +/- 5%, which was significantly greater than that seen in group II (distensibility = 4 +/- 2%, p < 0.001). In undilated vessels, distensibility was related to the degree of atherosclerotic burden (r = 0.75). This relation was curvilinear with a marked decrease in distensibility when percent plaque area exceeded 30%. Distensibility in group III (after PTCA) was higher than in group II (10 +/- 3 vs 4 +/- 2%, p < 0.001) despite a larger plaque burden (percent plaque area of 56 +/- 12 vs 46 +/- 11%, p < 0.005). The distensibility in group IV (after DCA) was also higher than in group II (8 +/- 4 vs 4 +/- 2%, p < 0.001) despite a similar residual percent plaque area (49 +/- 13 vs 46 +/- 11%, p = NS).(ABSTRACT TRUNCATED AT 250 WORDS)


Site specific diagnostic yield of endoscopic biopsies in Gastrointestinal Graft-versus-Host Disease: A tertiary care Center experience.
Gastrointestinal (GI) graft versus host disease (GVHD) occurs in up to 40% of patients undergoing allogenic hematopoietic stem cell transplantation (HSCT). However, the optimal endoscopic approach is still unclear and the area of the GI tract with the highest diagnostic yield is still a topic of debate. We compared the diagnostic yield of different anatomic site biopsies in the diagnosis of GI GVHD and assessed the correlation of endoscopic findings with histopathology. All cases of biopsy proven GI GVHD were obtained from pathology database AUBMC between 1/1/2005 and 31/8/2017. We retrospectively analyzed the demographical, clinical and endoscopic data. Nineteen patients were diagnosed with GI GVHD over 17.6 years. The most common presenting symptom was severe diarrhea (18 patients, 94.7%). Combining upper endoscopy and sigmoidoscopy with biopsies had the highest diagnostic yield of 90% in diagnosing GI GVHD compared to 63.6%, 78.6% and 77.8% for upper endoscopy, sigmoidoscopy and colonoscopy respectively. In macroscopically normal mucosa, the recto-sigmoid and duodenal biopsies had the highest diagnostic yield (75%). As for the macroscopically abnormal mucosa, the highest yield was for the recto-sigmoid biopsies (100%) in lower endoscopy and duodenal biopsies in the upper endoscopy (60%). In a patient suspected to have GI GVHD, the best endoscopic approach is the combination of upper endoscopy and flexible sigmoidoscopy with biopsies of normal as well as abnormal mucosa. It should be emphasized that normal mucosa be biopsied especially in the duodenum and recto-sigmoid for a better diagnostic yield.


Mortality in Yusho patients exposed to polychlorinated biphenyls and polychlorinated dibenzofurans: a 50-year retrospective cohort study.
In 1968, the Yusho incident resulted in accidental exposure to polychlorinated biphenyls (PCBs), polychlorinated dibenzofurans (PCDFs), and related compounds in Japan. This study updated the risk of mortality in Yusho patients. We obtained updated cohort data for all Yusho patients for the period 1968-2017. We calculated standardized mortality ratios (SMRs) for all-cause and cause-specific mortality over a 50-year follow-up period compared with the general population in Japan. A total of 1664 Yusho patients with 63,566 person-years of follow up were included in the analysis. Among males, excess mortality was observed for all cancers (SMR: 1.22, 95% confidence interval [CI]: 1.02 to 1.45) and lung cancer (SMR: 1.59, 95% CI: 1.12 to 2.19). Among females, increased mortality was observed for liver cancer (SMR: 2.05, 95% CI: 1.02 to 3.67). No significant increase was seen in non-cancer-related mortality compared with the general population. Carcinogenic risk in humans after exposure to PCBs and PCDFs remains higher among Yusho patients. Our findings suggest the importance of care engagement and optimum management to deal with the burden of Yusho disease.


A Feedback Loop of LINC00665 and the Wnt Signaling Pathway Expedites Osteosarcoma Cell Proliferation, Invasion, and Epithelial-Mesenchymal Transition.
Osteosarcoma (OS) is a malignant tumor with frequent occurrence among teenagers. Long non-coding RNAs (lncRNAs) play pro-cancer roles in many tumors. The purpose of this study was to figure out the functional role of a novel lncRNA long intergenic non-protein coding RNA 665 (LINC00665) in OS by observing the OS cell behaviors. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to analyze LINC00665 expression in OS cells. Cell function assays assessed the impacts of LINC00665 on OS cell phenotype. Immunofluorescence and western blot analyzed the function of LINC00665 on epithelial-mesenchymal transition (EMT) in OS. Moreover, mechanistic assays analyzed the downstream mechanism of LINC00665 in OS cells. LINC00665 was significantly up-regulated in OS cells. LINC00665 silence facilitated OS cell proliferation, migration, invasion, and EMT while inhibiting cell apoptosis. Mechanically, LINC00665 acted as a competing endogenous RNA (ceRNA) to sponge miR-1249-5p and thereby modulated Wnt family member 2B (WNT2B) to activate Wnt pathway. Wnt pathway activated LINC00665 expression transcriptionally. Our study uncovered the cancer-promoting role of LINC00665 in OS, and the feedback loop of LINC00665/miR-1249-5p/WNT2B/Wnt might be a potential target for OS treatment.


Early social enrichment affects responsiveness to different social cues in female mice.
Communal nesting (CN), an early social enrichment procedure in which multiple females rear the offspring in a single nest, increases maternal care levels received by offspring and interaction with peers. It has been shown that male mice reared under CN conditions show increased social competence and propensity to social interactions at adulthood. In the present study we investigated long-term behavioural effects of CN on female offspring. Mouse pups were reared under two different experimental conditions: standard nesting (SN, where single mother rears her pups) and CN (three females rearing their pups in a single nest). At adulthood CN and SN virgin females underwent three different behavioural tests: (i) maternal induction following presentation of foster pups; (ii) social recognition test in which ultrasound vocalizations (USVs) and social investigation behaviour emitted by a resident female in the presence of a female partner were recorded; (iii) zero-maze to analyze anxiety profiles. CN females showed (i) decreased licking response in the maternal induction test accompanied by an increased sniffing response; (ii) decreased of social interest towards a novel partner (during the Retest Different phase), and decreased USV emission rate in the social recognition test; CN and SN females did not differ in the emotional responses measured in the zero-maze apparatus. As a whole these data suggest that CN rearing render female mice less reactive to social novelty.


Lymphangiography, ultrasonography, and computed tomography in Hodgkin's disease and non-Hodgkin's lymphoma.
Findings from histologic analysis, lymphangiography, ultrasonography, and computed tomography were reviewed for 54 cases of Hodgkin's disease and 18 cases of non-Hodgkin's lymphoma. All patients were classified as clinical stage 1 or 2 disease at the time of the imaging studies. The ultrasound and computed tomography studies identified only 30 to 40% of the truly positive patients. This low sensitivity contrasts with lymphangiography, which identified 95% of the truly positive Hodgkin's disease patients and 70% of the patients with abdominal spread of non-Hodgkin's lymphoma. Many errors in interpretation were attributable to location and distribution of disease in these patients and the differing patterns of spread in Hodgkin's disease and non-Hodgkin's lymphoma. The evidence indicates that when findings are positive on computed tomography or ultrasound, no other study is necessary. In stage 1 or 2 lymphoma, non-Hodgkin's patients with negative noninvasive findings and all Hodgkin's patients, regardless of their noninvasive findings, should undergo lymphangiography.


Parameter identifiability and model selection for sigmoid population growth models.
Sigmoid growth models, such as the logistic, Gompertz and Richards' models, are widely used to study population dynamics ranging from microscopic populations of cancer cells, to continental-scale human populations. Fundamental questions about model selection and parameter estimation are critical if these models are to be used to make practical inferences. However, the question of parameter identifiability - whether a data set contains sufficient information to give unique or sufficiently precise parameter estimates - is often overlooked. We use a profile-likelihood approach to explore practical parameter identifiability using data describing the re-growth of hard coral. With this approach, we explore the relationship between parameter identifiability and model misspecification, finding that the logistic growth model does not suffer identifiability issues for the type of data we consider whereas the Gompertz and Richards' models encounter practical non-identifiability issues. This analysis of parameter identifiability and model selection is important because different growth models are in biological modelling without necessarily considering whether parameters are identifiable. Standard practices that do not consider parameter identifiability can lead to unreliable or imprecise parameter estimates and potentially misleading mechanistic interpretations. For example, using the Gompertz model, the estimate of the time scale of coral re-growth is 625 days when we estimate the initial density from the data, whereas it is 1429 days using a more standard approach where variability in the initial density is ignored. While tools developed here focus on three standard sigmoid growth models only, our theoretical developments are applicable to any sigmoid growth model and any appropriate data set. MATLAB implementations of all software are available on GitHub.


Eliminating brightness induction effects when measuring motion centre-surround suppression of contrast.
The perceived contrast of a central stimulus is supressed when it is embedded in a higher contrast surround, centre-surround suppression of contrast. Local brightness induction effects between the two stimulus regions have been proposed to account for conflicting results when relative grating phases were different. Here, suppression and brightness induction effects are dissociated using a centre-surround arrangement with moving gratings. Four experienced observers were involved in experiments, utilising two-interval forced-choice contrast matching tasks. The stimuli were drifting sinusoidal grating patterns with surrounds (95% contrast) differing in direction of motion and orientation relative to the 40% contrast centre grating. First a 90°-phase-offset same direction surround condition was compared to both same direction (phase aligned) and opposing direction conditions. The reduction in the suppression for the phase-offset condition suggested a reduction in brightness induction influences. Then suppression was examined when surround directions varied and where phase was either fixed or randomised. For small changes in the motion direction between centre and surround (0° to 26.6°) the amount of brightness induction varied sinusoidally with the difference in phase introduced by the direction difference. Finally, the spatial separation between the centre and surround was varied to determine the reduction of suppression and brightness induction with increasing spatial distance. We found both fit an exponential decay function, with surround suppression producing the larger range of influence. Our findings quantify both brightness induction and suppression effects and validate the use of phase randomisation to remove effects of brightness induction when evaluating surround suppression.


Low serum albumin level is associated with cytomegalovirus reactivation in patients with chronic lymphoproliferative diseases treated with alemtuzumab (Campath-1H)-based therapies.
Alemtuzumab (Campath-1H), a monoclonal antibody that targets the CD52 antigen, has been approved for the treatment of fludarabine-refractory chronic lymphocytic leukemia. However, the profound immunosuppression caused by alemtuzumab has been associated with infectious complications. The authors report on the incidence and risk factors for development of symptomatic cytomegalovirus reactivation in 113 patients with chronic lymphoproliferative disorders who received alemtuzumab-based therapy. Kaplan-Meier methods were applied to generate survival curves, and the log-rank test was used to assess the difference between groups; in addition, univariate and multivariate Cox proportional hazards models were used to estimate the hazard ratio of death including 95% confidence intervals. Cytomegalovirus reactivation was diagnosed in 25 patients (22%), and most of those patients responded to antiviral therapy. Nine additional patients had asymptomatic cytomegalovirus viremia. With appropriate therapy, most patients achieved clearance of cytomegalovirus viremia. Low serum albumin was the only factor associated significantly with symptomatic cytomegalovirus reactivation.


A volitional help sheet to reduce binge drinking in students: a randomized exploratory trial.
This study tested the ability of a volitional help sheet (VHS) to decrease binge drinking in UK students. Fifty-six participants were randomly allocated to one of three conditions: control, active control or VHS as part of a questionnaire-based study. There were significant decreases in units of alcohol consumed and self-reported binge drinking frequency in the VHS condition, but not in either of the control conditions. The findings support use of the VHS to help people to reduce their alcohol consumption and binge drinking.


Differences in In Vitro Properties of Pancreatin Preparations for Pancreatic Exocrine Insufficiency as Marketed in Russia and CIS.
Pancreatic enzyme-replacement therapy (PERT), provided as pancreatin to patients with pancreatic exocrine insufficiency (PEI), is considered an essential substitute for the pivotal physiological function the pancreas fulfills in digestion. PEI involves a reduction in the synthesis and secretion of pancreatic enzymes (lipase, protease, amylase), which leads to an inadequate enzymatic response to a meal and consequently to maldigestion and malabsorption of nutrients. The efficacy of PERT is strongly dependent on enzyme activity, dissolution, and pancreatin particle size. The physiological properties of eight pancreatin preparations (nine batches; five different brands) available in Russia and CIS (Commonwealth of Independent States: Armenia, Azerbaijan, Belarus, Kazakhstan, Kyrgyzstan, Moldova, Russia, Tajikistan, Uzbekistan) were investigated. The lipase activity, dissolution, and particle size distribution of samples from multiple batches of pancreatin of different strengths were measured. Regarding lipase activities, all pancreatin preparations except Micrazim® matched the labeled content. Considerable differences were observed in particle size and dissolution. Pancreatin preparations available in Russia and CIS demonstrate product-to-product and batch-to-batch variability regarding the measured properties of lipase activity, dissolution, and particle size. This may impact the efficacy of PERT and therefore clinical outcomes.


Systematic review of ultrasound gel associated Burkholderia cepacia complex outbreaks: Clinical presentation, sources and control of outbreak.
Burkholderia cepacia complex (Bcc) is an emerging opportunistic pathogen among immunocompromised patients. It frequently contaminates saline, fluids and ultrasound (US) gel used in hospitals. This systematic review was conducted to analyze Bcc outbreaks due to ultrasound (US) gel for better management of these outbreaks. As per PRISMA guidelines, electronic databases "Embase" and "Pubmed" and "Web of sciences" were searched from 1991 to April, 2021 to identify studies causing Burkholderia spp outbreak due to contamination of US gels. The search identified 14 outbreak reports that met our inclusion criteria. Bacteremia was the most common clinical presentation in ten studies followed by urinary tract infections in 4 studies. In most of the studies B. cepacia was the most common isolated organism. Other members like B. ambifaria, B. contaminans, and B. stabilis caused outbreaks in two studies. Pulsed field gel electrophoresis and multilocus sequence typing were commonly employed methods to study the clonal association. In 8 outbreaks, intrinsic contamination of the gel, that is, contamination from manufacturing site, was present and 4 studies, extrinsic contamination, that is, contamination from environment was responsible for outbreak. This review highlights the importance of US gel as a source of outbreak in health-care facilities. Ensuring sterility of US gel, sound epidemiological investigation of outbreak and prompt response by infection control team can prevent these outbreaks.


A systems biology preview of the relationships between mineral and metabolic complications in chronic kidney disease.
There are emerging data that the skeleton is connected to systemic biological functions through the release of two osteoblast-/osteocyte-derived hormones, fibroblastic growth factor 23 (FGF23) and undercarboxylated osteocalcin (Ocn). FGF23 is important in the regulation of phosphate and vitamin D metabolism, whereas Ocn participates in endocrine networks, coordinating bone and fat mass, energy metabolism, and sex hormone production. Bone remodeling and mineralization per se, along with the hormones leptin, insulin, glucocorticoids, PTH, and 1,25(OH)2D, regulate the release of FGF23 and Ocn, leading to complex cross-talk and coordination between endocrine networks previously thought to be distinct. These pathways are particularly important in chronic kidney disease, in which both FGF23 and Ocn are increased. Although these hormones initially serve an adaptive role, with progressive loss of renal function they show maladaptive effects, particularly on the cardiovascular system, through multiple mechanisms, including possible cross-talk with the renin angiotensin system. The complex interconnections between the various endocrine networks in chronic kidney disease may account for the difficulty in treating the uremic state.


Rhythms in human bone marrow and blood cells.
In 24h studies of bone marrow (BM), circadian stage-dependent variations were demonstrated in the proliferative activity of BM cells from subsets of 35 healthy diurnally active men. On an average, the percentage of total BM cells in deoxyribonucleic acid (DNA) synthesis phase was 188% greater at midday than at midnight (circadian rhythm: p = 0.018; acrophase or peak time of 13: 16h). Patients with malignant disease (n = 15) and a normal cortisol circadian rhythm showed higher fractions of BM cells in S-phase at midday. Colony-forming units--granulocyte/macrophage (CFU-GM), an indicator of myeloid progenitor cells, showed the same circadian variation as DNA S-phase (average range of change or ROC = 136%; circadian rhythm: p < 0.001; acrophase of 12:09h). Deoxyribonucleic acid S-phase and CFU-GM in BM both showed a circannual rhythm (p = 0.015 and 0.008) with an identical acrophase of August 12. The daily peak in BM glutathione content, a tripeptide involved in cellular defense against cytotoxic damage, preceded BM proliferative peaks by 4-5 h (ROC = 31-90%; circadian rhythm: p = 0.05; acrophase of 08:30h). Myeloid (ROC = 57%; circadian rhythm: p = 0.056; acrophase at 08:40h) and erythroid (ROC = 26%; circadian rhythm: p = 0.01; acrophase of 13:01h) precursor cells were positively correlated (r = 0.41; p < 0.001), indicating a circadian temporal relationship and equal influence on S-phase of total BM cells. Yield of positive selected CD34+ progenitor stem cells also showed significant circadian variation (ROC = 595%; circadian rhythm: p = 0.02; acrophase of 12:40h). Thus, the temporal synchrony in cell cycling renders BM cells more sensitive at specific times to hematopoietic growth factors and cell cycle-specific cytotoxic drugs. Moreover, proper timing of BM harvesting may improve progenitor cell yield. When using marker rhythms in the blood to allow for individualized timing of BM procedures, the times of low values in white blood corpuscles, neutrophils, and lymphocytes and high values in cortisol were predictive of the times of highest BM erythroid, myeloid, and total S-phase numbers occurring in the following 12 h.


EuCAP, a Eukaryotic Community Annotation Package, and its application to the rice genome.
Despite the improvements of tools for automated annotation of genome sequences, manual curation at the structural and functional level can provide an increased level of refinement to genome annotation. The Institute for Genomic Research Rice Genome Annotation (hereafter named the Osa1 Genome Annotation) is the product of an automated pipeline and, for this reason, will benefit from the input of biologists with expertise in rice and/or particular gene families. Leveraging knowledge from a dispersed community of scientists is a demonstrated way of improving a genome annotation. This requires tools that facilitate 1) the submission of gene annotation to an annotation project, 2) the review of the submitted models by project annotators, and 3) the incorporation of the submitted models in the ongoing annotation effort. We have developed the Eukaryotic Community Annotation Package (EuCAP), an annotation tool, and have applied it to the rice genome. The primary level of curation by community annotators (CA) has been the annotation of gene families. Annotation can be submitted by email or through the EuCAP Web Tool. The CA models are aligned to the rice pseudomolecules and the coordinates of these alignments, along with functional annotation, are stored in the MySQL EuCAP Gene Model database. Web pages displaying the alignments of the CA models to the Osa1 Genome models are automatically generated from the EuCAP Gene Model database. The alignments are reviewed by the project annotators (PAs) in the context of experimental evidence. Upon approval by the PAs, the CA models, along with the corresponding functional annotations, are integrated into the Osa1 Genome Annotation. The CA annotations, grouped by family, are displayed on the Community Annotation pages of the project website http://rice.tigr.org, as well as in the Community Annotation track of the Genome Browser. We have applied EuCAP to rice. As of July 2007, the structural and/or functional annotation of 1,094 genes representing 57 families have been deposited and integrated into the current gene set. All of the EuCAP components are open-source, thereby allowing the implementation of EuCAP for the annotation of other genomes. EuCAP is available at http://sourceforge.net/projects/eucap/.


Lymphocyte response to mitogens in patients with malignant melanoma.
Lymphocyte blast transformation of 23 melanoma patients was compared to that of 22 healthy persons after stimulation with the mitogens PHA, ConA, and PWM. Transformation rate of lymphocytes in microcultures was measured following 3H-Thymidin uptake in a Liquid-Scintillation-Counter. Calculations were based on analyses of variance. There was no significant difference in blastogenetic response between the 2 groups (patients and controls), but there were differences between the used mitogens.


Axonal damage in the optic radiation assessed by white matter tract integrity metrics is associated with retinal thinning in multiple sclerosis.
White matter damage in the visual pathway is common in multiple sclerosis (MS) and is associated with retinal thinning, although the underlying mechanism of association remains unclear. The goal of this work was to evaluate the presence and extent of white matter tract integrity (WMTI) alterations in the optic radiation (OR) in people with MS and to investigate the association between WMTI metrics and retinal thinning in the eyes of MS patients without a history of optic neuritis (ON) as measured by optical coherence tomography (OCT). We hypothesized that WMTI metrics would reflect axonal damage that occurs in the OR in MS, and that axonal alterations revealed by WMTI would be associated with retinal thinning. Twenty-nine MS patients without previous ON in at least one eye and twenty-nine age-matched healthy controls (HC) were scanned on a dedicated high-gradient 3-Tesla MRI scanner with 300 mT/m maximum gradient strength using a multi-shell diffusion MRI protocol (b = 800, 1500, 2400 s/mm2). The patients were divided into two subgroups according to history without ON (N = 18) or with ON in one eye (N = 11). Diffusion tensor imaging (DTI) metrics and WMTI metrics derived from diffusion kurtosis imaging were assessed in normal-appearing white matter (NAWM) of the OR and in focal lesions. Retinal thickness in the eyes of MS patients was measured by OCT. Student's t-test was used to assess group differences between MRI metrics. Linear regression was used to study the relationship between OCT metrics, including retinal nerve fiber layer (RNFL) and combined ganglion cell and inner plexiform layer thickness (GCL/IPL), visual acuity measures and DTI and WMTI metrics. OR NAWM in MS showed significantly decreased axonal water fraction (AWF) compared to HC (0.36 vs 0.39, p < 0.001), with similar trends observed in AWF of lesions compared to NAWM (0.27 vs 0.36, p < 0.001). Fractional anisotropy (FA) was lower in OR NAWM of MS patients compared to HC (0.49 vs 0.52, p < 0.001). In patients without ON, AWF was the only diffusion MRI metric that was significantly associated with average RNFL (r = 0.68, p = 0.005), adjusting for age, sex and disease duration and correcting for multiple comparisons. Of all the DTI and WMTI metrics, AWF was the strongest and most significant predictor of average RNFL thickness in MS patients without ON. There was no significant correlation between visual acuity scores and DTI or WMTI metrics after correction for multiple comparisons. Axonal damage may be the substrate of previously observed DTI alterations in the OR, as supported by the significant reduction in AWF within both NAWM and lesions of the OR in MS. Our results support the concept that axonal damage is widespread throughout the visual pathway in MS and may be mediated through trans-synaptic degeneration.


Nicotinic acetylcholine receptors in the rat stomach: II. Alterations after vagotomy.
Nicotinic and muscarinic acetylcholine receptors were studied in the stomach of vagotomized rats, with ligand binding techniques, and with the concomitant measurement of choline acetyltransferase (ChAT). The maximum binding capacity (Bmax) of the high affinity sites of (-)-[3H]nicotine binding was significantly increased in all the regions of the stomach of subdiaphragmatically vagotomized animals while the values for the low affinity sites remained the same as those in sham-operated animals. On the other hand, [3H]quinuclidinyl benzilate binding to the stomach was unchanged after vagotomy. ChAT activity was significantly reduced in the vagotomized rats. These results suggest that surgical vagal denervation causes an increase in the density of nicotinic acetylcholine receptors presumably located on the parasympathetic ganglion cells. There is also a reduction of ChAT activity, probably due to a degenerative loss of preganglionic cholinergic neurons. Preganglionic denervation (decentralization) has little influence on the muscarinic receptors of postsynaptic effector organs.


Description of the immature stages of four species of Macrodactylini (Coleoptera: Melolonthidae: Melolonthinae).
The third instar of the genera Ceraspis, Clavipalpus, Isonychus and Manopus (Melolonthidae: Melolonthinae: Macrodactylini) are described for the first time. Descriptions are based on the larvae of Ceraspis innotata (Blanchard), Clavipalpus ursinus Blanchard, Isonychus maculatus Waterhouse and Manopus biguttatus Laporte. The pupae of C. ursinus, I. maculatus and M. biguttatus are also described. A key to the larvae of nine genera of Macrodactylini and a list of the species with immature descriptions are provided.


Process analytical technology as in-process control tool in semi-continuous manufacturing of PLGA/PEG-PLGA microspheres.
Nowadays, among 3rd generation drug delivery systems, biodegradable polymeric based long-acting injectable depot has achieved tremendous success in clinical application. So far, there have been two dozen of commercial products of Poly (lactic-co-glycolic acid) microspheres available in the market. Recently, continuous manufacturing concept has been successfully applied on oral solid formulation from buzzword to reality. However, the polymeric injectable microspheres are still stayed at batch manufacturing phase due to the lack of understanding of knowledge matrix. In this study, micro-mixer as a plug-and-play emulsification modules, Raman spectroscopy and focused beam reflectance measurement as real-time monitoring modules are integrated into a novel semi-continuous manufacturing streamline to provides more efficient upscaling flexibility in microspheres production. In this end to end semi-continuous manufacturing process, amphiphilic block polymer monomethoxy-poly (ethylene glycol) modified PLGA (mPEG-PLGA) was used for encapsulating Gallic acid. Additionally, with guarantee of good robustness, the correlation relationship between critical process parameters, critical material attributes and critical quality attributes were investigated. The time-space evolution process and mechanism for formation of PEG-PLGA microsphere with particular morphology were elaborated. Altogether, this study firstly established semi-continuous manufacturing streamline for PLGA/PEG-PLGA microspheres, which would not only lower the cost of production, narrow process variability and smaller equipment/environmental footprint but also applied in-process control (IPC) and QbD principle on complicated production process of microspheres. Therefore, this study build confidence in the industrial development of PLGA/PEG-PLGA microspheres and establish best practice standards, which might be a quantum leap for developing PLGA microspheres in the future.


Comparison of four current 25-hydroxyvitamin D assays.
The performance of recently developed vitamin D total assays (ADVIA Centaur and Elecsys) was compared to that of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and LIASON 25-OH Vitamin D total assays. A total of 157 clinical samples and standard reference material (SRM) 972 were analyzed. The correlations of LC-MS/MS with the three immunoassays were acceptable. However, compared to LC-MS/MS, LIAISON and ADVIA Centaur showed negative bias, and Elecsys showed positive bias. There was a lack of agreement among the four methods with only LC-MS/MS results close to the certified values of SRM 972. The prevalence of vitamin D insufficiency (<50 nmol/L) was higher with ADVIA Centaur (51.6%) and LIAISON (52.2%) and lower with Elecsys (37.6%), compared with that of LC-MS/MS (44.6%). The new, automated total vitamin D assays show acceptable correlation with LC-MS/MS, and could be used in routine laboratories. However, standardization of vitamin D assays and consideration of assay-specific decision limits should be addressed.


Genetic diversity and molecular markers of the tropical abalone (Haliotis asinina) in Thailand.
Genetic diversity of abalone in Thailand, Haliotis asinina, H. ovina, and H. varia, was analyzed by polymerase chain reaction (PCR) of 18S and 16S rDNAs, with randomly amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP). Species-specific RAPD markers were found in each abalone species. Restriction analysis of 18S (nuclear) ribosomal DNA with Alu I, Taq I, and Hae III and 16S (mitochondrial) rDNA with Bam HI, Eco RI, Hae III, and Alu I gave 12 and 13 digestion patterns, respectively. A total of 49 composite haplotypes were found. A dendogram obtained by the unweighted pair-group method with arithmetic mean, constructed from divergence between pairs of composite haplotypes, revealed reproductively isolated gene pools of these abalone and indicated that H. asinina and H. ovina are genetically closer than H. varia. When H. varia was discovered owing to small sample sizes, geographic heterogeneity analysis and FST estimate indicated clear genetic differentiation between H. ovina originating from the Andaman Sea (west) and the Gulf of Thailand (east, P<0.0001), whereas partial differentiation was observed between the Philippines and the remaining H. asinina samples (P<0.0021). The amplified 16S rDNAs of individuals representing composite haplotypes found in this study were cloned and sequenced. A neighbor-joining tree constructed from sequence divergence of 16S rDNA accurately allocated those sequences according to species origins of abalone. Species-specific PCR based on 16S rDNA polymorphism was successfully developed in H. asinina and H. varia but not in H. ovina.


Ultrastructural changes in the rat metrial gland in the latter half of pregnancy.
The ultrastructural changes occurring in the metrial gland in the latter half of pregnancy in the rat have been studied. Typical palely stained granulated cells are present in the metrial gland up to day 20 but many granulated cells show variations in appearance which may be associated with degeneration. In some the cytoplasm is more darkly stained and such cells often have apparently empty areas of cytoplasm adjacent to the granules. From day 14 onwards many areas of the gland show cellular debris, apparently resulting from lysis of the granulated cells. However, occasionally normal granulated cells are present in blood vessels and have been observed apparently penetrating the vascular endothelium. A variety of changes was also noted in the stromal cell population. Inclusions became apparent in many of these cells; some of these consisted of lipid while others resembled granules from the typical granulated cells. Other cells with numerous inclusions appeared to be macrophages. Lysis of granulated cells in situ is compatible with suggestions that the metrial gland produces a holocrine secretion, though some normal granulated cells enter blood vessels. The stromal cells may have a phagocytic role in late pregnancy but evidence for this was inconclusive.


Systematic meta-review and quality assessment of the structural brain alterations in schizophrenia.
The large quantity of systematic reviews of magnetic resonance imaging studies in schizophrenia challenges their meaningful interpretation. This meta-review synthesises the available information from systematic reviews of structural alteration in both chronic and first-episode schizophrenia. Systematic reviews were identified using electronic databases. Review methodological quality was assessed according to the Assessment of Multiple Systematic Reviews checklist. Data were extracted in duplicate and quality assessed for consistency and precision, guided by Grading of Recommendations Assessment, Development and Evaluation recommendations. Integration of volumetric and voxel-based estimates allowed critical assessment of the magnitude and location of anatomical differences. There is evidence for grey matter reductions of anterior cingulate, frontal (particularly medial and inferior) and temporal lobes, hippocampus/amygdala, thalamus, and insula that may be magnified over time. Other regional alterations appear specific to illness stage or medication status. There is limited high quality evidence supporting grey or white matter changes in schizophrenia, which has previously been obscured by a large volume of conflicting lower quality evidence.


In vivo analysis of chromatin following nystatin-mediated import of active enzymes into Saccharomyces cerevisiae.
In vivo DNA-protein interactions are usually studied at the molecular level using DNA-degrading agents of low molecular weight. In order to be useful, macromolecular probes of chromatin structure, such as enzymes must first cross the cell membrane. In this paper we describe the introduction and evaluation of macromolecules with enzymatic activity into yeast spheroplasts treated with the polyene antibiotic nystatin. We report the low resolution analysis of chromatin structure in the promoter region of the Saccharomyces cerevisiae gene encoding DNA topoisomerase I by this technique using micrococcal nuclease and restriction enzymes.


Association of SRD5A2 variants and serum androstane-3alpha,17beta-diol glucuronide concentration in Chinese elderly men.
Results of recent studies have demonstrated that genetic variants of the enzyme steroid 5α reductase type II (SRD5A2) are associated with serum concentrations of major androgen metabolites such as conjugates of androstane-3α,17β-diol-glucuronide (3α-diol-G). However, this association was not consistently found among different ethnic groups. Thus, we aimed to determine whether the association with SRD5A2 genetic variations exists in a cohort of healthy Chinese elderly men, by examining 2 metabolite conjugates: androstane-3α,l7β-diol-3-glucuronide (3α-diol-3G) and androstane-3α,17β-diol-17-glucuronide (3α-diol-17G). We used GC-MS and LC-MS to measure serum sex steroid concentrations, including testosterone and dihydrotestosterone, and 3α-diol-3G and 3α-diol-17G in 1182 Chinese elderly men age 65 and older. Genotyping of the 3 SRD5A2 tagSNPs [rs3731586, rs12470143, and rs523349 (V89L)] was performed by using melting-temperature-shift allele-specific PCR. The well-described SRD5A2 missense variant rs523349 (V89L) was modestly associated with the 3α-diol-17G concentration (P = 0.040). On the other hand, SNP rs12470143 was found to be significantly correlated with 3α-diol-3G concentration (P = 0.021). Results of haplotype analysis suggested that the presence of an A-C-G haplotype leads to an increased 3α-diol-3G concentration, a finding consistent with results of single SNP analysis. The genetic variation of SRD5A2 is associated with circulating 3α-diol-3G and 3α-diol-17G concentrations in Chinese elderly men. In addition, we showed that SRD5A2 haplotypic association, rather than a single SNP alone, might be a better predictor of the 3α-diol-G concentration. Thus, the effect of either the haplotype itself or of other ungenotyped SNPs in linkage disequilibrium with the haplotype is responsible for the interindividual variation of 3α-diol-G.


A novel myxozoan parasite of terrestrial mammals: description of Soricimyxum minuti sp. n. (Myxosporea) in pygmy shrew Sorex minutus from Hungary.
As part of a biodiversity study in northwestern Hungary, we conducted a parasitological survey of small mammals. In both common shrews (Sorex araneus Linnaeus) and pygmy shrews (Sorex minutus Linnaeus), we found myxospores of a species of Soricimyxum Prunescu, Prunescu, Pucek et Lom, 2007 (Myxosporea) and plasmodia in the bile ducts within the liver. Spores from both species of shrewswere morphologically and morphometrically indistinguishable, but differed in their SSU rRNA gene sequences by 3.3%. We identified spores and developmental stages from the common shrew as Soricimyxum fegati Prunescu, Prunescu, Pucek et Lom, 2007, based on morphometric data and DNA sequence similarity. Spores from the pygmy shrew were only 96.7% similar to S. fegati, hence we identified them as a novel myxosporean Soricimyxum minuti sp. n. This is only the second myxosporean parasite species described from mammals.


A mathematical model of coronavirus transmission by using the heuristic computing neural networks.
In this study, the nonlinear mathematical model of COVID-19 is investigated by stochastic solver using the scaled conjugate gradient neural networks (SCGNNs). The nonlinear mathematical model of COVID-19 is represented by coupled system of ordinary differential equations and is studied for three different cases of initial conditions with suitable parametric values. This model is studied subject to seven class of human population N(t) and individuals are categorized as: susceptible S(t), exposed E(t), quarantined Q(t), asymptotically diseased IA (t), symptomatic diseased IS (t) and finally the persons removed from COVID-19 and are denoted by R(t). The stochastic numerical computing SCGNNs approach will be used to examine the numerical performance of nonlinear mathematical model of COVID-19. The stochastic SCGNNs approach is based on three factors by using procedure of verification, sample statistics, testing and training. For this purpose, large portion of data is considered, i.e., 70%, 16%, 14% for training, testing and validation, respectively. The efficiency, reliability and authenticity of stochastic numerical SCGNNs approach are analysed graphically in terms of error histograms, mean square error, correlation, regression and finally further endorsed by graphical illustrations for absolute errors in the range of 10-05 to 10-07 for each scenario of the system model.


Treatment of gastric adenocarcinoma at a New Zealand centre: meeting the challenges of a low volume country.
Surgical treatment of gastric cancer in New Zealand is challenging because of a low annual incidence of 400 patients and population dispersal over a wide area with little data on regional treatment trends and outcomes. This investigation was undertaken to evaluate the surgical outcomes of gastric cancer patients presenting to a single upper gastrointestinal centre (WDHB, Waitemata District Health Board) and to compare these to national and international standards. Data on 135 patients with histologically proven gastric adenocarcinoma presenting between January 2010 and December 2014 were reviewed and compared with nationally available procedural volume data. Sixty of 135 patients were resected (resection rate 44%) and 75 patients were managed with palliative chemotherapy/symptomatic care alone. Twenty-six patients (43%) received adjuvant or neoadjuvant chemotherapy and only two patients (3%) underwent laparoscopic resection. In resected patients, 90-day mortality was 1.6%, and 11 patients (18%) developed complications ≥ Clavien-Dindo grade 3. Fifty-two patients (87%) had ≥15 lymph nodes resected and 54 patients (90%) had a histological R0 resection. At median follow-up of 49 months, 30 patients remain alive and disease-free with 20 true 5-year disease-free survivors. National data between 2010 and 2014 showed WDHB performed 20% (338/1710) of gastric resections for all indications in New Zealand. While WDHB is an internationally low volume centre for gastric cancer, surgical outcomes benchmark satisfactorily to international standards. New Zealand's national treatment standards should set aspirational goals for gastric cancer treatment and have a clear strategy to address issues of surgical volume and national service provision.


Enriched experience and recovery from amblyopia in adult rats: impact of motor, social and sensory components.
Amblyopia is one of the most common forms of visual impairment, arising from an early functional imbalance between the two eyes. It is currently accepted that, due to a lack of neural plasticity,amblyopia is an untreatable pathology in adults. Environmental enrichment (EE) emerged as a strategy highly effective in restoring plasticity in adult animals, eliciting recovery from amblyopia through a reduction of intracortical inhibition. It is unknown whether single EE components are able to promote plasticity in the adult brain, crucial information for designing new protocols of environmental stimulation suitable for amblyopic human subjects. Here, we assessed the effects of enhanced physical exercise,increased social interaction, visual enrichment or perceptual learning on visual function recovery in adult amblyopic rats. We report a complete rescue of both visual acuity and ocular dominance in exercised rats, in animals exposed to visual enrichment and in animals engaged in perceptual learning.These effects were accompanied by a reduced inhibition/excitation balance in the visual cortex. In contrast, we did not detect any sign of recovery in socially enriched rats or in animals practicing a purely associative visual task. These findings could have a bearing in orienting clinical research in the field of amblyopia therapy.


[The control of Aujeszky's disease using surface vaccination. 2. Serologic studies of the occurrence of latent infection by Aujeszky's disease virus in fattening swine].
The aim of the study was to prove in a region in the North-West of Germany that there is the possibility to decrease the prevalence of latent Aujeszky's Disease infections by means of a systematic vaccination. For this, the most important precondition of the success in a high population immunity that is maintained also in the finishing pig until slaughter. Since it is known that maternal antibodies interfere with the active immunization in the prefattening period, procedures were to be found that guarantee a high immunity in spite of the colostral antibodies. The results of the study show that the existing recommendation for vaccinating fattening pigs are sufficient for the prevalence of latent infections. They also confirm the necessity to carry out a check- and slaughter policy at the end of the eradication program.


Variability in eukaryotic initiation factor iso4E in Brassica rapa influences interactions with the viral protein linked to the genome of Turnip mosaic virus.
Plant potyviruses require eukaryotic translation initiation factors (eIFs) such as eIF4E and eIF(iso)4E to replicate and spread. When Turnip mosaic virus (TuMV) infects a host plant, its viral protein linked to the genome (VPg) needs to interact with eIF4E or eIF(iso)4E to initiate translation. TuMV utilizes BraA.eIF4E.a, BraA.eIF4E.c, BraA.eIF(iso)4E.a, and BraA.eIF(iso)4E.c of Brassica rapa to initiate translation in Arabidopsis thaliana. In this study, the BraA.eIF4E.a, BraA.eIF4E.c, BraA.eIF(iso)4E.a, and BraA.eIF(iso)4E.c genes were cloned and sequenced from eight B. rapa lines, namely, two BraA.eIF4E.a alleles, four BraA.eIF4E.c alleles, four BraA.eIF(iso)4E.a alleles, and two BraA.eIF(iso)4E.c alleles. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) analyses indicated that TuMV VPg could not interact with eIF4E, but only with eIF(iso)4E of B. rapa. In addition, the VPgs of the different TuMV isolates interacted with various eIF(iso)4E copies in B. rapa. In particular, TuMV-UK1/CDN1 VPg only interacted with BraA.eIF(iso)4E.c, not with BraA.eIF(iso)4E.a. Some single nucleotide polymorphisms (SNPs) were identified that may have affected the interaction between eIF(iso)4E and VPg such as the SNP T106C in BraA.eIF(iso)4E.c and the SNP A154C in VPg. Furthermore, a three-dimensional structural model of the BraA.eIF(iso)4E.c-1 protein was constructed to identify the specific conformation of the variable amino acids from BraA.eIF(iso)4E.c. The 36th amino acid in BraA.eIF(iso)4E.c is highly conserved and may play an important role in establishing protein structural stability. The findings of the present study may lay the foundation for future investigations on the co-evolution of TuMV and eIF(iso)4E.


Lipid metabolic vulnerabilities of multiple myeloma.
Multiple myeloma (MM) is the second most common hematological malignancy worldwide, characterized by abnormal proliferation of malignant plasma cells within a tumor-permissive bone marrow microenvironment. Metabolic dysfunctions are emerging as key determinants in the pathobiology of MM. In this review, we highlight the metabolic features of MM, showing how alterations in various lipid pathways, mainly involving fatty acids, cholesterol and sphingolipids, affect the growth, survival and drug responsiveness of MM cells, as well as their cross-talk with other cellular components of the tumor microenvironment. These findings will provide a new path to understanding the mechanisms underlying how lipid vulnerabilities may arise and affect the phenotype of malignant plasma cells, highlighting novel druggable pathways with a significant impact on the management of MM.


Correction to: Sebaceous carcinoma of trunk with bilateral axillary lymph node metastasis: a rare presentation of malignant adnexal tumor in young adult.
[This corrects the article DOI: 10.1093/jscr/rjac280.].


Detection of encephalomyocarditis virus in clinical samples by immunomagnetic separation and one-step RT-PCR.
A method of immunomagnetic separation and one-step reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of Encephalomyocarditis virus (EMCV). EMCV was captured from sample on magnetic beads with homologous monoclonal antibody and then heat denatured. The heated beads were used directly in one-step RT-PCR reaction to amplify a 285-bp PCR fragment at the 3' end of the genomic region that encodes the viral polymerase. This method detected as little as 3.5 TCID(50) of EMCV from infected cell culture. It was shown with this method that the sensitivity of RT-PCR increased when applied for the detection of EMCV added to fecal extract. Using this protocol EMCV was detected from heart homogenate samples containing less than 100 TCID(50)/ml. The amplified product was sequenced to ensure specificity. The immunomagnetic-RT/PCR procedure described here should be useful for the rapid, specific and sensitive detection of EMCV in clinical samples. This technique is rapid, reliable and can be readily adapted to detect EMCV from other clinical samples.


Subunit stoichiometry of the Drosophila melanogaster small nuclear RNA activating protein complex (SNAPc).
Small nuclear RNA activating protein complex (SNAPc) is a multi-subunit transcription factor required for expression of small nuclear RNA genes. This protein binds to a promoter element located approximately 40-65 bp upstream of the transcription start site. In Drosophila melanogaster, DmSNAPc contains three distinct polypeptide subunits: DmSNAP190, DmSNAP50, and DmSNAP43. The subunit stoichiometry in SNAPc complexed with DNA has not been examined. Therefore, the ability of differently tagged but otherwise identical subunits to associate with each other into the same protein-DNA complex was assayed by antibody super-shift analysis. The results reveal that DmSNAPc contains only a single copy of each of the three subunits.


Involution of thymic epithelium and low serum thymulin bioactivity in weanling mice subjected to severe food intake restriction or severe protein deficiency.
The volume of epithelium in the cortex and in the medulla of the thymus was compared in four groups of weanling male and female CBA/J mice. Well-nourished controls (C), food intake restricted (R), and animals given a low-protein diet ad libitum (LP) were fed from 23 to 37 days of age. Baseline controls (B) were studied at 23 days of age. Epithelial volume fraction was estimated for each group by point-counting morphometry on electron micrographs. Other mice were used to obtain group mean estimates of thymic index (mg/g live weight) and volume fraction of cortex and medulla (light microscope-level point-counting morphometry). Cortical and medullary epithelial volumes were calculated for each animal examined by electron microscopy by obtaining the live weight and applying, in sequence, the group mean thymic index, an assumed thymic density of 1.0 mm3/mg, the group mean cortical or medullary volume fraction, and the measured cortical or medullary volume fraction for that animal. Serum thymulin bioactivity was also measured in C, R, and LP mice. The results reveal thymic epithelial involution in the two most common rodent models of malnutrition, and suggest that this may contribute to the low serum thymulin levels found in malnourished experimental animals and humans.


1H NMR-based metabolomic study of the effects of flavonoids on citrinin production by Monascus.
Monascus comprises purple-red molds. Various compounds can be obtained from these species, including statins and food-safe yellow, red, and orange pigments. However, the secondary metabolite citrinin, a mycotoxin, is produced during the late stages of growth. Citrinin biosynthesis should be reduced to apply Monascus pigments safely. Fortunately, this can be achieved by the addition of flavonoids (genistein, daidzein, apigenin, and kaempferol). However, the effects of these flavonoids on other metabolites remain unknown. Here, we report a 1H NMR-based multivariate metabolomic analysis of the effects of flavonoids on mycotoxin citrinin production by Monascus. Fifteen metabolites involved in lysine and arginine biosynthesis and alanine, aspartate, glutamate, biotin, arginine, proline, and glutathione metabolism were detected. The reduction in glutamate, aspartate, biotin, and 2-phosphoglycerate content suggested their association with the citrinin reduction mechanism. This study identifies the citrinin production pathway in Monascus and will aid in the development of citrinin-control methods.


Transcriptionally-active high-risk human papillomavirus is rare in oral cavity and laryngeal/hypopharyngeal squamous cell carcinomas--a tissue microarray study utilizing E6/E7 mRNA in situ hybridization.
Human papillomavirus is well established in oropharyngeal squamous cell carcinoma as both causative and prognostic, but its significance in non-oropharyngeal tumours is unclear. In particular, the significance of finding viral DNA is not known. We sought to evaluate nonoropharyngeal squamous cell carcinomas for transcriptionally-active human papillomavirus and to compare this with the presence of viral DNA. We evaluated an 87 patient tissue microarray cohort of oral cavity and laryngeal/hypopharyngeal squamous cell carcinomas for high risk human papillomavirus DNA and E6 and E7 mRNA transcripts by in situ hybridization, and for p16 expression by immunohistochemistry. We found only two of the 73 (2.7%) evaluable cases to harbour transcriptionally-active human papillomavirus. Both of these tumours were from the larynx, one was positive for human papillomavirus DNA by in situ hybridization, and both were extensively positive for p16. All oral cavity and hypopharyngeal tumours were negative for human papillomavirus. Transcriptionally-active human papillomavirus appears to be rare in laryngeal, hypopharyngeal, and oral cavity squamous cell carcinomas. As such, it appears unlikely to be a 'driver' or to be clinically significant in most established tumours.


The effects of various nutritional supplements on the growth, migration and differentiation of Xenopus laevis neural crest cells in vitro.
This study investigates the nutritional requirements of Xenopus laevis neural crest cells and melanophores developing in vitro. A comparison is made between the growth and differentiation of cells in serum-containing medium and a chemically defined, serum-free medium that we have designed. Our chemically defined medium is more efficient than serum-supplemented medium in promoting proliferation of these cells. Several supplements are required to enhance culture development. These include insulin, alpha-melanocyte stimulating hormone, somatotropin, luteotrophic hormone, linoleic acid, uridine, and putrescine. In addition, collagen and fibronectin provide the most conducive environment tested for cell migration and adhesion.


Influence of gut microbiota on mucosal IgA antibody response to the polio vaccine.
The impact of intestinal microbiota on mucosal antibody response to the polio vaccine is poorly understood. We examined changes in vaccine-induced intestinal mucosal immunity to poliovirus by measuring the immunoglobulin A (IgA) antibody levels in stool samples collected from 107 infants in China, and the samples were collected 14 days after different sequential vaccinations combining inactivated polio vaccine (IPV) with oral poliovirus vaccine (OPV). Gut microbiota were identified using 16S ribosomal RNA sequencing 28 days before, 14 days before, and at the last dose of OPV. Vaccine-induced type 2-specific mucosal IgA showed a decrease after switching from trivalent to bivalent OPV (bOPV) (positive rate of polio type 2-specific mucosal IgA, 16.7%, 11.8%, and 45.9% for IPV + 2bOPV, 2IPV + bOPV, and 2IPV + trivalent OPV groups, respectively). The composition of the gut microbiome was significantly different, a higher abundance of Firmicutes and a lower abundance of Actinobacteria were observed in IgA-negative infant (n = 66) compared with IgA-positive infants (n = 39), and the gut microbiota were more diverse in IgA-negative infants on the day of OPV inoculation. The abundance of Clostridia was concomitant with a significantly lower conversion rate of mucosal IgA responses to the polio vaccine. The composition of the gut microbiome may affect the intestinal mucosal IgA response to the polio vaccine.


Rounding of low serum creatinine levels and consequent impact on accuracy of bedside estimates of renal function in cancer patients.
To compare glomerular filtration rate measured by technetium-99m ([Tc(99m)]) DTPA clearance with estimated creatinine clearance (CrCl) (Cockcroft and Gault (C&G) method) in patients with serum creatinine (Scr) levels <0.06 mmol l(-1), and determine the effect of rounding serum creatinine to 0.06 mmol l(-1). Patients with serum creatinine values <0.06 mmol l(-1) at the time of [Tc(99m)] clearance determination were identified. Creatinine clearance was calculated by the C&G method using both actual and rounded Scr values. A total of 419 adults had GFR measured by technetium-99m diethyl triamine penta-acetic acid ([Tc(99m)] DTPA) clearance. Out of this group, 26 patients had a serum creatinine value <0.06 mmol l(-1). The C&G estimates of renal function using actual serum creatinine resulted in an overall overestimation of 12.9% when compared to [Tc(99m)] DTPA clearance. When the value of serum creatinine was rounded to 0.06 mmol l(-1), the formula underestimated renal function by -7.0%. Analysis of estimated creatinine clearance for different levels of renal function showed significant differences to [Tc(99m)] DTPA clearance. Rounding up of serum creatinine to 0.06 mmol l(-1) improved the predictive ability of the C&G method for the patients with [Tc(99m)] DTPA clearance </=100 ml min(-1), but worsened the effect in those >100 ml min(-1). This work indicates that when bedside estimates of renal function are calculated using the C&G formula actual Scr should be used first to estimate CrCl. If the resultant CrCl is </=100 ml min(-1), then the Scr should be rounded up to 0.06 mmol l(-1) and CrCl recalculated. Further assessment of this approach is warranted in a larger cohort of patients.


Smoking is an important factor that affects peripheral blood progenitor cells yield in healthy male donors.
Smoking could reduce the CD34+ cells in peripheral blood of healthy individual. This study aimed to investigate the correlation between smoking load and the effect of peripheral blood hematopoietic progenitor cells (PBPCs) mobilization by granulocyte colony-stimulating factor (G-CSF) alone in healthy donors. Retrospective analysis was performed on 145 healthy adult PBPCs donors who underwent PBPCs mobilization and collection. Smoking factors were evaluated and correlated with mobilization responses, as indicated by the collected CD34+ cells concentration. The collected CD34+ cells concentration was closely related to pre-CD34 (P < .001) and CD34+ cells collected per volume blood processed (P < .001) which suggested that collected CD34+ cells concentration was a reliable indicator of PBPCs mobilization efficiency. The heavy smoking donors revealed significantly lower collected CD34+ cells concentration, compared to that of the nonsmoking (P < .001) and light smoking donors (P < .05). The levels of collected CD34+ cells in light smoking were also obviously lower than that in nonsmoking donors (P < .05).There were no obvious differences in the collected CD34+ cells concentration, overall processed blood volume and total collected CD34+ cells between nonsmoking and smoking cessation groups (P = .490; P = .464; P = .819). Cigarette smoking is an important factor that affects the yield of PBPCs in male donors, especially when the smoking load is more than five pack-years. Mobilization of PBMCs could be restored by smoking cessation in chronic smokers.


Porcine spermadhesin AQN-3 binds to negatively charged phospholipids.
The spermadhesin AQN-3 is a major component of porcine seminal plasma. While various studies suggest that this protein binds to boar sperm cells, its attachment to the cells is poorly understood. Therefore, the capacity of AQN-3 to interact with lipids was investigated. For that purpose, AQN-3 was recombinantly expressed in E. coli and purified via the included His-tag. Characterizing the quaternary structure by size exclusion chromatography revealed that recombinant AQN-3 (recAQN-3) is largely present as multimer and/or aggregate. To determine the lipid specificity of recAQN-3, a lipid stripe method and a multilamellar vesicle (MLV)-based binding assay were used. Both assays show that recAQN-3 selectively interacts with negatively charged lipids, like phosphatidic acid, phosphatidylinositol phosphates, and cardiolipin. No interaction was observed with phosphatidylcholine, sphingomyelin, phosphatidylethanolamine, or cholesterol. The affinity to negatively charged lipids can be explained by electrostatic interactions because binding is partly reversed under high-salt condition. However, more factors have to be assumed like hydrogen bonds and/or hydrophobic forces because the majority of bound molecules was not released by high salt. To confirm the observed binding behavior for the native protein, porcine seminal plasma was incubated with MLVs comprising phosphatidic acid or phosphatidyl-4,5-bisphosphate. Attached proteins were isolated, digested, and analyzed by mass spectrometry. Native AQN-3 was detected in all samples analyzed and was - besides AWN - the most abundant protein. It remains to be investigated whether AQN-3, together with other sperm associated seminal plasma proteins, acts as decapacitation factor by targeting negative lipids with signaling or other functional roles in fertilization.