Epidermal growth factor receptor plays an anabolic role in bone metabolism in vivo.
While the epidermal growth factor receptor (EGFR)-mediated signaling pathway has been shown to have vital roles in many developmental and pathologic processes, its functions in the development and homeostasis of the skeletal system has been poorly defined. To address its in vivo role, we constructed transgenic and pharmacologic mouse models and used peripheral quantitative computed tomography (pQCT), micro-computed tomography (µCT) and histomorphometry to analyze their trabecular and cortical bone phenotypes. We initially deleted the EGFR in preosteoblasts/osteoblasts using a Cre/loxP system (Col-Cre Egfr(f/f)), but no bone phenotype was observed because of incomplete deletion of the Egfr genomic locus. To further reduce the remaining osteoblastic EGFR activity, we introduced an EGFR dominant-negative allele, Wa5, and generated Col-Cre Egfr(Wa5/f) mice. At 3 and 7 months of age, both male and female mice exhibited a remarkable decrease in tibial trabecular bone mass with abnormalities in trabecular number and thickness. Histologic analyses revealed decreases in osteoblast number and mineralization activity and an increase in osteoclast number. Significant increases in trabecular pattern factor and structural model index indicate that trabecular microarchitecture was altered. The femurs of these mice were shorter and smaller with reduced cortical area and periosteal perimeter. Moreover, colony-forming unit-fibroblast (CFU-F) assay indicates that these mice had fewer bone marrow mesenchymal stem cells and committed progenitors. Similarly, administration of an EGFR inhibitor into wild-type mice caused a significant reduction in trabecular bone volume. In contrast, Egfr(Dsk5/+) mice with a constitutively active EGFR allele displayed increases in trabecular and cortical bone content. Taken together, these data demonstrate that the EGFR signaling pathway is an important bone regulator and that it primarily plays an anabolic role in bone metabolism.


Differential carbohydrate expression in tumorigenic vs nontumorigenic prostate cell-lines.
Death from prostate cancer is most frequently a result of metastatic disease. A key step in the process of metastasis is the attachment of circulatory tumor cells to target organ endothelium. This process is thought to be mediated by lectins, a class of cell surface proteins that bind two or more carbohydrate groups. Using fluorescent microscopy and fluorescein isothiocyanate (FITC) conjugated lectins, the presence of various carbohydrates was examined in the following tumorigenic and non-tumorigenic cell lines: rat prostate epithelial (EPYP-2, EPYP-1), rat prostate endothelial (YPEN-2, YPEN-1, YPEN-2PV), Dunning rat prostate cancer cell lines (MLL, ML, AT6.3, AT.1, AT2.1, GP9F3), and three tumorigenic human prostate cell lines (LNCaP, PC3, PC3 bone). The lectin Triticum vulgaris (WGA) was found to readily bind the carbohydrates N-acetylglucosamine and sialic acid on the plasma membrane of tumorigenic cell lines. Interestingly, WGA bound carbohydrates located to the nucleus and cytoplasm in non-tumorigenic cell lines, indicating that N-acetylglucosamine and sialic acid residues are preferentially expressed on the cell membrane of prostate cancer cells. Lectin staining patterns in cell lines of varying metastatic potential revealed no significant difference between highly metastatic vs. low metastatic cell lines. Observations revealed an absence of N-acetylgalactosamine and L-fucose in all rat Dunning, epithelial and endothelial cell lines as well as all human prostate cancer cell lines except for the androgen insensitive human prostate cancer cell line PC3, in which L-fucose residues were detected in the nucleus and on the plasma membrane. PC3 bone cells, which metastasized selectively to bone, demonstrated the presence of galactose residues whereas PC3 cells did not, suggesting that preference for target organ endothelium may be influenced by the expression of carbohydrate residues. These data indicate that differential carbohydrate expression may play an important role in prostate cancer biology.


Induction of a putative monooxygenase of crabs (Carcinus spp.) by polycyclic aromatic hydrocarbons.
As part of a programme to develop biomarker assays for polycyclic aromatic hydrocarbons (PAHs) in marine invertebrates, two species of crabs, Carcinus maenas and Carcinus aestuarii were exposed to benzo(a)pyrene (B(a)P) or crude oil. Microsomes were prepared from the midgut gland (hepatopancreas), examined by gel electrophoresis and Western blotting and assayed for B(a)P monooxygenase activity. In early experiments there was evidence of protein degradation and results were inconsistent and inconclusive. However, when steps were taken to minimize this in subsequent experiments, including the inclusion of four protease inhibitors in the homogenization buffer, there was consistent evidence for an increase of proteins of estimated molecular weight 45-60 kDa, and particularly of a distinct band at c. 48 kDa, following exposure to PAH at levels down to 0.1 ppm in ambient water. In C. aestuarii the increase in this band was found to coincide with an 8-12-fold increaseof B(a)P monooxygenase activity in midgut gland microsomes. These results suggest that one or more forms of cytochrome P450 may be induced by PAHs in these species. However, Western blotting using antibodies raised to vertebrate P450s, and representing four different gene families, failed to recognize any proteins in either the PAH-treated samples or in the controls. The isolation and characterization of induced protein, and the production of antibodies may provide the basis for a biomarker assay to measure a response to environmental PAHs in crabs.


Management of atmospheric pollutants from waste incineration processes: the case of Bozen.
This article presents the case study of a waste incinerator located in a region rich in natural and environmental resources, and close to the city of Bozen, where there are about 100,000 inhabitants. Local authorities paid special attention to the effect of the plant on human health and the surrounding environment. Indeed, among the measures adopted to control the emissions, in 2003 an automatic sampling system was installed specifically to monitor polychlorinated dibenzo-p-dioxin and polychlorinated dibenzofuran (PCDD/F) emissions during the complete operation time of the plant. The continuous sampling system was coupled directly to aerosol spectrometers for the determination of fine and ultra-fine particles in the emissions of the plant. The measurement results suggest that the waste incineration plant of Bozen is not a significant source of PCDD/F, or fine and ultra-fine particles. Immission measurements from other monitoring systems confirmed these results.


Cryptic speciation and limited hybridization within Lumbricus earthworms (Clitellata: Lumbricidae).
Cryptic mitochondrial (mt) lineages are known to exist in the earthworm morphospecies Lumbricus rubellus and L. terrestris. The latter was recently split into two species, L. terrestris and L. herculeus, based on large genetic distances and a statistical difference in body size. There is support for the separation of some lineages in L. rubellus into species, whereas other lineages, separated by similar mt genetic distances, have been found to be part of the same species. However, no study has evaluated the status of the cryptic mt lineages in L. terrestris-L. herculeus and L. rubellus using nuclear genes. We use a combination of methods to reveal extensive cryptic speciation and limited hybridization in Lumbricus, based on one nuclear (H3) and one mitochondrial (COI) marker. Using a Bayesian multi-locus species delimitation method, as well as single gene haplotype networks and gene trees, we delimit seven well supported cryptic species within the morphospecies L. rubellus, and confirm the split within the species-pair L. terrestris-L. herculeus. Limited hybridization was found between the most common species of L. rubellus (A) in northern Europe and two other species (B and H) in this complex, as well as between L. terrestris and L. herculeus. Deep mt divergence was found within L. terrestris s.str. but no support for further splitting of this taxon was found. Both L. rubellus and L. terrestris are well studied model organisms, and considering that cryptic species and hybridization were found within them, it is important that they are properly identified in future studies.


A Semiparametric Bayesian Approach to Epidemics, with Application to the Spread of the Coronavirus MERS in South Korea in 2015.
We consider incomplete observations of stochastic processes governing the spread of infectious diseases through finite populations by way of contact. We propose a flexible semiparametric modeling framework with at least three advantages. First, it enables researchers to study the structure of a population contact network and its impact on the spread of infectious diseases. Second, it can accommodate short- and long-tailed degree distributions and detect potential superspreaders, who represent an important public health concern. Third, it addresses the important issue of incomplete data. Starting from first principles, we show when the incomplete-data generating process is ignorable for the purpose of Bayesian inference for the parameters of the population model. We demonstrate the semiparametric modeling framework by simulations and an application to the partially observed MERS epidemic in South Korea in 2015. We conclude with an extended discussion of open questions and directions for future research.


Conversion of finished leather waste incorporated with plant fibers into value added consumer products - An effort to minimize solid waste in Ethiopia.
Presently, the leftovers from leather product industries are discarded as waste in Ethiopia. The objective of the present study was therefore, to prepare composite sheets by incorporating various plant fibers like enset (Ensete ventricosum), hibiscus (Hibiscus cannabinus), jute (Corchorus trilocularis L.), palm (Phoenix dactylifera) and sisal (Agave sisal) in various proportions into the leather waste. Resin binder (RB) and natural rubber latex (NRL) were used as binding agents for the preparation of the composite sheets. The composite sheets prepared were characterized for their physicochemical properties (tensile strength, elongation at break, stitch tear strength, water absorption, water desorption and flexing strength). Composite sheets prepared using RB having 10% hibiscus, 20% palm and 40% sisal fibers showed better mechanical properties than their respective controls. In composite sheets prepared using NRL having 30% jute fiber exhibited better mechanical properties than its control. Most of the plant fibers used in this study played a role in increasing the performance of the sheets. However, as seen from the results, the contribution of these plant fibers on performance of the composite sheets prepared is dependent on the ratio used and the nature of binder. The SEM studies have exhibited the composite nature of the sheets and FTIR studies have shown the functional groups of collagen protein, cellulose and binders. The prepared sheets were used as raw materials for preparation of items like stiff hand bags, ladies' purse, keychain, chappal upper, wallet, wall cover, mouse pad and other interior decorating products. By preparing such value added products, we can reduce solid waste; minimize environmental pollution and thereby securing environmental sustainability.


Barriers to and Facilitators of a Career as a Physician-Scientist Among Rheumatologists in the US.
To determine perceived barriers to and facilitators of a career in rheumatology research, examine factors leading rheumatologists to leave an academic research career, and solicit ways to best support young physician-scientists. A web-based survey was conducted among the domestic American College of Rheumatology (ACR) membership from January through March 2014. Inclusion criteria were ACR membership and an available e-mail address. Non-rheumatologists were excluded. The survey assessed demographics, research participation, barriers to and facilitators of a career in research, reasons for leaving a research career (when applicable), and ways in which the ACR could support junior investigators. Content analysis was used to extract relevant themes. Among 5,448 domestic ACR members, 502 responses were obtained (9.2% response rate). After exclusions (38 incomplete, 2 duplicates, 32 non-rheumatologists), 430 responses were analyzed. Participants included fellows, young investigators, established investigators, mentors, clinicians, and those who previously pursued a research career but have chosen a different career path. Funding and mentoring were the most highly ranked barriers and facilitators. Protection from clinical and administrative duties, institutional support, and personal characteristics such as resilience and persistence were also ranked highly. The most commonly cited reasons for leaving an academic research career were difficulty obtaining funding and lack of department or division support. This is the first study to examine barriers to and facilitators of a career in rheumatology research from the perspectives of diverse groups of rheumatologists. Knowledge of such barriers and facilitators may assist in designing interventions to support investigators during vulnerable points in their career development.


Cross-linking of IgGs bound on circulating neutrophils leads to an activation of endothelial cells: possible role of rheumatoid factors in rheumatoid arthritis-associated vascular dysfunction.
Rheumatoid arthritis is characterized by the presence of circulating auto-antibodies, including rheumatoid factors, which recognize the Fc portion of IgGs. The neutrophil is the most abundant circulating leukocyte and it expresses high levels of FcγRs on its surface. The aim of the present study was to examine the capacity of circulating human neutrophils to be activated by rheumatoid factors and the consequences of these events on endothelium. Neutrophil-bound IgGs were cross-linked with anti-human IgGs to mimick the presence of circulating rheumatoid factors and FcγRs-dependent signalling events and functions were examined. The IgG and IgM composition of rheumatoid factors isolated from the serum of RA patients was characterized. Adhesion of neutrophils to endothelial cells was quantified in response to the addition of rheumatoid factors. Cross-linking of IgGs bound on neutrophils leads to FcγRs-dependent tyrosine phosphorylation, mobilisation of intracellular calcium and the extracellular release of superoxide anions and lysozyme. Incubation of endothelial cells with the supernatant of activated neutrophils increases ICAM-1 expression and IL-8 production by endothelial cells. Finally, rheumatoid factors enhance neutrophil adhesion to endothelial cells. Our results show that activation of neutrophils' FcγRs by rheumatoid factors could participate in rheumatoid arthritis-associated vascular damage.


Propolis as an Adjuvant in the Healing of Human Diabetic Foot Wounds Receiving Care in the Diagnostic and Treatment Centre from the Regional Hospital of Talca.
Diabetic foot wounds are a relevant diabetes complication and a major health problem. It has been described that propolis has health benefits due to its anti-inflammatory, antioxidant, and support in the healing process. The current study assessed the effect of propolis as an adjuvant in the healing of human diabetic foot ulcers. This was evaluated in a randomized placebo-controlled study of subjects receiving care in the Diagnostic and Treatment Centre from the Regional Hospital of Talca, Chile. Randomized subjects received ambulatory healing treatment for diabetes foot wounds with propolis spray (3%), which was applied to cover the entire wound surface each time it was dressed from week 0 until cicatrization or 8 weeks as a maximum. Two serum samples were taken (day 0 and end of the study) for cytokine and oxidative stress analyses. Also, macro- and microscopy were analyzed in the process of wound healing. The study comprised 31 subjects with type 2 diabetes in treatment for diabetic foot wounds in the Diagnostic and Treatment Centre from the Regional Hospital of Talca. Propolis promotes a reduction of the wound's area by an average of 4 cm2, related to an increase in the connective tissue deposit compared to the control. Also, propolis increased the glutathione (GSH) and GSH/glutathione disulfide (GSSG) ratio (p < 0.02), depleted tumor necrosis factor- (TNF-) α, and increased interleukin- (IL-) 10 levels. Topical propolis did not modify the biochemical parameters in the serum of the studied subjects. The topical use of propolis turned out to be an interesting therapeutic strategy as an adjuvant in the care of diabetes foot wounds due to its ability to improve and promote healing based on its anti-inflammatory and antioxidant profile. This trial is registered with NCT03649243.


Fast motion estimation using bidirectional gradient methods.
Gradient-based motion estimation methods (GMs) are considered to be in the heart of state-of-the-art registration algorithms, being able to account for both pixel and subpixel registration and to handle various motion models (translation, rotation, affine, and projective). These methods estimate the motion between two images based on the local changes in the image intensities while assuming image smoothness. This paper offers two main contributions. The first is enhancement of the GM technique by introducing two new bidirectional formulations of the GM. These improve the convergence properties for large motions. The second is that we present an analytical convergence analysis of the GM and its properties. Experimental results demonstrate the applicability of these algorithms to real images.


[Treating coronary heart disease by acupuncture at neiguan (PC6) and xinahu (BL15): an efficacy assessment by SPECT].
To evaluate the clinical efficacy of acupuncture combined single photon emission computed tomography (SPECT) in treating coronary heart disease (CHD) and its effect on the myocardial ischemia/perfusion and the recovery of heart functions. Totally fifty-nine patients with confirmed CHD were randomly assigned to two groups, the acupuncture group (32 cases) and the nitroglycerine group (27 cases). Patients in the acupuncture group were electro-acupunctured at bilateral Neiguan (PC6) and Xinshu (BL15) for 30 min with the frequency of 2/15 Hz and the current strength 9 - 18 mA after myocardial imaging induced by routine exercises or drug load. 99mTc-MIBI 370 MBq was injected 15 min after needling. The myocardial perfusion imaging was performed immediately after needling. 99mTc-MIBI740 MBq was injected to those in the nitroglycerine group during routine exercises or drug load. The myocardial perfusion imaging was performed 5 min after injection. Patients were asked to sublingual administration of nitroglycerine 1 mg after the myocardial perfusion imaging was completed. 99mTc-MIBI 370 MBq was intravenously injected 5 min later, and myocardial perfusion imaging was performed 5 min after injection. There was statistical difference in changes of radioactive uptake between before and after treatment in the two groups (P < 0.05, P < 0.01). Both acupuncture and buccal administration of nitroglycerine could increase the blood perfusion of ischemic myocardium. But there was no statistical difference in the improvement of ischemic myocardial cells (t = 1.57, P > 0.05). Using SPECT could clearly display therapeutic effects of acupuncture on CHD, thus providing a new visible research method for CHD studies.


Deaf versus hearing subjects' recall of words on a distraction task as a function of the signability of the words.
Subjects, including 17 deaf and 10 hearing students in Grades 11 and 12, completed a test for memory of lists of 6 words (presented visually for 10 sec.). Subjects recalled the words in writing after a distracting task of adding pairs of digits for 10 sec. Word lists are categorized as signable with a single sign, compound or combination of signs, or finger-spelling signs only. Hearing subjects recalled significantly more words in each category than did deaf subjects. Deaf subjects recalled significantly more single-signed words than either of the other two categories. Deaf subjects did not recall more compound/combination signed words than words that could only be finger-spelled. Hearing subjects also recalled significantly more single-signed words than either of the other two categories and were not superior in either of the other categories.


Cholestane-3beta, 5alpha, 6beta-triol promotes vascular smooth muscle cells calcification.
Oxysterols found in atherosclerotic plaque may be associated with vascular calcification. We investigated the effect of oxysterol cholestane-3beta, 5alpha, 6beta-triol (Triol) on in vitro calcification of rat vascular smooth muscle cells (VSMCs). In vitro calcification was induced by incubation of VSMCs with beta-glycerophosphate. Calcifying nodule formation, calcium deposition in extracellular matrix, and alkaline phosphatase (ALP) activity were measured as indices of calcification. Because apoptotic bodies can serve as nucleation sites for calcification, apoptosis of calcifying VSMCs was determined by Hoechst 33258 staining, TUNEL, and FITC-labeled annexin V/PI double staining. The calcium deposition and ALP activity in calcifying VSMCs were much higher than those in non-calcifying VSMCs. Triol increased calcifying nodule formation, calcium deposition, ALP activity, and apoptosis of nodular cells in calcifying VSMCs. As determined by 2,7-dichlorofluorescein fluorescence, Triol induced the generation of reactive oxygen species (ROS) in calcifying VSMCs dose- and time-dependently. Triol-induced increases in calcium deposition, ALP activity, apoptosis, and ROS generation were all attenuated by antioxidant vitamin C plus vitamin E (VC + VE). The results demonstrated that Triol promoted VSMCs calcification through direct increase of ALP activity and apoptosis, probably by ROS-related mechanism.


A clinic-based pilot intervention to enhance diabetes management for elderly Hispanic patients.
Successful diabetes self-management requires behavioral and lifestyle changes. However, low-income patients may face challenges related to poverty that make it difficult to engage in lifestyle changes. We piloted an intervention designed to help older, low-income, Hispanic, patients with diabetes access free or low-cost community resources to enhance diabetes self-management. Results from this pilot intervention are reported. Patients were recruited at baseline to complete surveys assessing diabetes self-care activities, diabetes self-efficacy, and general self-efficacy. Volunteers were trained by a clinic social worker to help patients identify needs and make referrals to local community resources (e.g., housing, transportation, food, clothing, dental and prescription services, employment, or family social services). Identical surveys were administered at 3-month follow-up. 28 patients were recruited at baseline and 18 patients completed the follow-up assessment. No significant changes in diabetes care and self-efficacy were detected. All patients requested at least one referral to a community resource. The most common requests were for low-cost dental clinics, food assistance, and housing support. At follow-up, nine (50%) patients contacted their given referrals. The need for assistance with basic social services is high in this population. The rate of referral uptake (50%) is high for a relatively low intensity intervention. Since the completion of the pilot, the program has trained 21 volunteers and helped over 220 patients in a primary care clinic. Using a volunteer model and creating connections to existing community resources is a cost-conscious way to deliver needed services to patients.


Out-of-pocket payments and community-wide health outcomes: an examination of influenza vaccination subsidies in Japan.
While studies have shown that reductions in out-of-pocket payments for vaccination generally encourages vaccination uptake, research on the impact on health outcomes has rarely been examined. Thus, the present study, using municipal-level survey data on a subsidy programme for influenza vaccination in Japan that covers the entire country, examines how reductions in out-of-pocket payments for vaccination among non-elderly individuals through a subsidy programme affected regional-level influenza activity. We find that payment reductions are negatively correlated with the number of weeks with a high influenza alert in that region, although the correlation varied across years. At the same time, we find no significant correlation between payment reductions and the total duration of influenza outbreaks (i.e. periods with a moderate or high alert). Given that a greater number of weeks with a high alert indicates a severer epidemic, our findings suggest that reductions in out-of-pocket payments for influenza vaccination among the non-elderly had a positive impact on community-wide health outcomes, indicating that reduced out-of-pocket payments contributes to the effective control of severe influenza epidemics. This suggests that payment reductions could benefit not only individuals by providing them with better access to preventive care, as has been shown previously, but also communities as a whole by shortening the duration of epidemics.


GrabBlur--a framework to facilitate the secure exchange of whole-exome and -genome SNV data using VCF files.
Next Generation Sequencing (NGS) of whole exomes or genomes is increasingly being used in human genetic research and diagnostics. Sharing NGS data with third parties can help physicians and researchers to identify causative or predisposing mutations for a specific sample of interest more efficiently. In many cases, however, the exchange of such data may collide with data privacy regulations. GrabBlur is a newly developed tool to aggregate and share NGS-derived single nucleotide variant (SNV) data in a public database, keeping individual samples unidentifiable. In contrast to other currently existing SNV databases, GrabBlur includes phenotypic information and contact details of the submitter of a given database entry. By means of GrabBlur human geneticists can securely and easily share SNV data from resequencing projects. GrabBlur can ease the interpretation of SNV data by offering basic annotations, genotype frequencies and in particular phenotypic information - given that this information was shared - for the SNV of interest. GrabBlur facilitates the combination of phenotypic and NGS data (VCF files) via a local interface or command line operations. Data submissions may include HPO (Human Phenotype Ontology) terms, other trait descriptions, NGS technology information and the identity of the submitter. Most of this information is optional and its provision at the discretion of the submitter. Upon initial intake, GrabBlur merges and aggregates all sample-specific data. If a certain SNV is rare, the sample-specific information is replaced with the submitter identity. Generally, all data in GrabBlur are highly aggregated so that they can be shared with others while ensuring maximum privacy. Thus, it is impossible to reconstruct complete exomes or genomes from the database or to re-identify single individuals. After the individual information has been sufficiently "blurred", the data can be uploaded into a publicly accessible domain where aggregated genotypes are provided alongside phenotypic information. A web interface allows querying the database and the extraction of gene-wise SNV information. If an interesting SNV is found, the interrogator can get in contact with the submitter to exchange further information on the carrier and clarify, for example, whether the latter's phenotype matches with phenotype of their own patient.


Risk factor profile and achievement of treatment goals among hypertensive patients from the Israeli Blood Pressure Control (IBPC) program--initial cost utility analysis.
Blood pressure (BP) reduction is crucial in reducing cardiovascular morbidity and mortality. The IBPC (Israeli Blood Pressure Control) program was initiated in order to enhance the control of modifiable risk factors among high-risk hypertensive patients under follow-up by general practitioners in Israel. The cost effectiveness of an intervention program is an important factor in the decision-making process of its implementation and therefore was evaluated here. The objective of this evaluation is to estimate the costs, monetary savings and benefits in terms of QALYs (quality-adjusted life years) that would be expected if the program were to be expanded to 100 clinics nationwide, enabling around 14800 persons to be treated. Hypertensive patients were screened in 30 general practice clinics, supervised by specialists in family medicine, each seeing 1000-5000 patients; 50-250 hypertensive patients were diagnosed at each participating clinic. BP levels, body mass index (BMI), lipid and glucose levels, as well as target organ damage and medications were recorded for all patients. A total of 4948 (2079, 42% males) were registered. Mean age was 64.8 +/- 12 years. After 1 year of follow-up versus baseline, the various parameters were as follows: BP control was achieved in 46.4% vs 29% of all hypertensive patients. LDL control (JNC VI criteria) was achieved in 41.7% vs 31.2% of all patients. Fasting plasma glucose control (glucose < 126 mg/dl) was achieved in 22% vs 19% of diabetic patients and 5.2% vs 3.1% of the diabetics had fasting plasma glucose levels > 200 mg/dl. Obesity (BMI > 30 kg/m2) was noted in 36.7% vs 43.8% at baseline. The cost utility analysis of the reduction in risk factors was calculated based on the international dicta applied to the reduction in risk factors as a result of treatment. For 100 clinics nationwide and 14800 persons to be treated the net saving to health services would be $977993 and the increase in QALYs would be 602 years. Better risk factor control in hypertensive patients by general practitioners could reduce morbidity and mortality as well as be cost effective.


The endothelial protein C receptor and malaria.
In this issue of Blood, Moxon et al provide novel insight into the pathogenesis of cerebral malaria, linking loss of the endothelial protein C receptor (EPCR) on brain vessels, caused by cytoadherent infected erythrocytes, with localized coagulation, inflammation, and disruption of endothelial barrier function.


Gender Differences in the Association of Brain Gray Matter and Pain-Related Psychosocial Characteristics.
Although the association of gray matter morphology alterations and pain-related psychosocial characteristics with pain intensity and chronification in people with chronic spinal pain is evident, research on their mutual interaction is scarce and does not account for possible gender differences. Gender-based differences are, however, of utmost importance to consider when examining pain neurobiology. To look for gender differences in the association between magnetic resonance imaging- (MRI) derived brain gray matter morphology and self-reported psychosocial characteristics. An explorative, observational study. University Hospitals Ghent and Brussels, Belgium. Brain gray matter morphology (using MRI) and self-reported psychosocial characteristics were examined in women and men with nonspecific chronic spinal pain. Statistical analyses were performed in SPSS and R to identify differences between men and women regarding brain gray matter, self-reported psychosocial characteristics, as well as gender differences in the association between those outcome measures. A total of 94 people with chronic spinal pain were studied, including 32 men (15 suffering from neck pain, 17 suffering from low back pain; demographics [mean ± SD] age: 45.00 ± 12.02 years; pain duration: 128.37 ± 110.45 months), and 62 women (36 suffering from neck pain, 26 suffering from low back pain; demographics [mean ± SD] age: 38.78 ± 12.69 years; pain duration: 114.27 ± 92.45 months). Woman showed larger (positive) associations of several central brain areas (paracentral, precentral, postcentral, etc.) with perceived consequences (P < 0.001), emotional representations (P < 0.001), chronicity (P < 0.001), and pain catastrophizing (P< 0.001). Men showed larger (both positive and negative) associations of the precuneus cortex, the precentral gyrus, and the insula with perceived personal control (P < 0.001) and kinesiophobia (P < 0.001). Other factors, such as menstrual cycle and medication can have a certain influence, and were only partly taken into consideration in the present investigation to obtain sufficient power. Another limitation is the observational study design, which hampers the possibility to look for causal or temporal interactions. Gray matter morphology relates differently to psychosocial characteristics in women and men. These explorative findings provide ideas for further research to investigate if targeting perceived negative consequences of the illness, perceived emotional representations, perceived chronicity, and pain catastrophizing in women, and perceived personal control of the illness and kinesiophobia in men, could contribute to the normalization of brain alterations in people with nonspecific chronic spinal pain. Gray matter, brain morphology, central nervous system, illness perceptions, central sensitization.


Anatomopathological study of dementia syndrome linked with an abnormal cerebrospinal fluid flow. Report of literature and personal observations.
The authors have analysed the data of the literature to identify the cases of normotensive hydrocephalus that underwent surgery and then died after the operation; some of these patients died over varying periods of time after the operation and the death was due to accidents. It seems that the anatomopathological lesions are less important in cases that benefited from the operation compared with cases that did not present perceptable clinical variations. The authors report the anatomopathological data of four personal cases which, from the clinical point of view, presented the dementia symptom associated in varying degrees to other neurological symptoms such as disturbances of the gait and of the sphincters functions (Adams' triad). All four subjects presented dilatation of the cerebral ventricles without cortical atrophy. From the histological point of view, there was: exfoliation of the ependyma, subependymal gliosis, demyelination of the white periventricular matter and spongiosis; there were no lesions of the meninges, of the cerebral cortex, no vascular alterations, except for those due to age, or stenosis of the aqueduct. The cause of the ventricular dilatation that was responsible for the clinical symptoms was not clear from the histological examination; the value and the significance of the histopathological data obtained and from the data available from the review of the literature are discussed and they point out the fact that many of the lesions encountered seem to be the consequence rather than the cause of the hydrocephalus.(ABSTRACT TRUNCATED AT 250 WORDS)


Separation of fast from slow anabolism by site-specific PEGylation of insulin-like growth factor I (IGF-I).
Insulin-like growth factor I (IGF-I) has important anabolic and homeostatic functions in tissues like skeletal muscle, and a decline in circulating levels is linked with catabolic conditions. Whereas IGF-I therapies for musculoskeletal disorders have been postulated, dosing issues and disruptions of the homeostasis have so far precluded clinical application. We have developed a novel IGF-I variant by site-specific addition of polyethylene glycol (PEG) to lysine 68 (PEG-IGF-I). In vitro, this modification decreased the affinity for the IGF-I and insulin receptors, presumably through decreased association rates, and slowed down the association to IGF-I-binding proteins, selectively limiting fast but maintaining sustained anabolic activity. Desirable in vivo effects of PEG-IGF-I included increased half-life and recruitment of IGF-binding proteins, thereby reducing risk of hypoglycemia. PEG-IGF-I was equipotent to IGF-I in ameliorating contraction-induced muscle injury in vivo without affecting muscle metabolism as IGF-I did. The data provide an important step in understanding the differences of IGF-I and insulin receptor contribution to the in vivo activity of IGF-I. In addition, PEG-IGF-I presents an innovative concept for IGF-I therapy in diseases with indicated muscle dysfunction.


Deep learning enables automatic detection and segmentation of brain metastases on multisequence MRI.
Detecting and segmenting brain metastases is a tedious and time-consuming task for many radiologists, particularly with the growing use of multisequence 3D imaging. To demonstrate automated detection and segmentation of brain metastases on multisequence MRI using a deep-learning approach based on a fully convolution neural network (CNN). Retrospective. In all, 156 patients with brain metastases from several primary cancers were included. 1.5T and 3T. [Correction added on May 24, 2019, after first online publication: In the preceding sentence, the first field strength listed was corrected.] SEQUENCE: Pretherapy MR images included pre- and postgadolinium T1 -weighted 3D fast spin echo (CUBE), postgadolinium T1 -weighted 3D axial IR-prepped FSPGR (BRAVO), and 3D CUBE fluid attenuated inversion recovery (FLAIR). The ground truth was established by manual delineation by two experienced neuroradiologists. CNN training/development was performed using 100 and 5 patients, respectively, with a 2.5D network based on a GoogLeNet architecture. The results were evaluated in 51 patients, equally separated into those with few (1-3), multiple (4-10), and many (>10) lesions. Network performance was evaluated using precision, recall, Dice/F1 score, and receiver operating characteristic (ROC) curve statistics. For an optimal probability threshold, detection and segmentation performance was assessed on a per-metastasis basis. The Wilcoxon rank sum test was used to test the differences between patient subgroups. The area under the ROC curve (AUC), averaged across all patients, was 0.98 ± 0.04. The AUC in the subgroups was 0.99 ± 0.01, 0.97 ± 0.05, and 0.97 ± 0.03 for patients having 1-3, 4-10, and >10 metastases, respectively. Using an average optimal probability threshold determined by the development set, precision, recall, and Dice score were 0.79 ± 0.20, 0.53 ± 0.22, and 0.79 ± 0.12, respectively. At the same probability threshold, the network showed an average false-positive rate of 8.3/patient (no lesion-size limit) and 3.4/patient (10 mm3 lesion size limit). A deep-learning approach using multisequence MRI can automatically detect and segment brain metastases with high accuracy. 3 Technical Efficacy Stage: 2 J. Magn. Reson. Imaging 2020;51:175-182.


Effect of prostanoids and their precursors on the aggregation of rainbow trout thrombocytes.
The role of prostanoids and their precursor fatty acids in the aggregatory response of thrombocytes (platelet equivalents of fish) from the rainbow trout, Oncorhynchus mykiss, was studied. Aggregation of these cells was induced by the thromboxane mimetic U-46619 or arachidonic acid (AA) in the presence of human or trout fibrinogen. The production of TXB2/3 by thrombocytes in response to stimulation with AA was inhibited by aspirin, ibuprofen, and indomethacin. However, thrombocyte aggregation in response to AA stimulation was not significantly altered by these agents at the concentrations tested (10-100 microM), with the exception of indomethacin at 20 and 40 microM. Effects on cytosolic calcium concentration have been suggested as an alternative mechanism for the inhibitory action of indomethacin on human platelet aggregation. The present study, however, failed to identify this as a mechanism for the inhibition of U-46619-induced trout thrombocyte aggregation by indomethacin. The polyunsaturated fatty acids docosahexaenoic acid and eicosapentaenoic acid both exhibited an inhibitory effect on U-46619-induced thrombocyte aggregation similar to that observed with mammalian platelets. Unlike the case in mammalian hemostasis, prostacyclin inhibited thrombocyte aggregation only at high concentrations (>5 microM). Prostaglandin E2, however, inhibited thrombocyte aggregation at much lower concentrations (>0.01 microM), suggesting that it may be the major inhibitory eicosanoid in trout.


Experimental determination of the alpha and beta C--H bond dissociation energies in naphthalene.
The acidities of the two different sites in naphthalene (1alpha and 1beta) and the electron affinities of the alpha- and beta-naphthyl radicals were measured using a Fourier transform mass spectrometer. Both carbon-hydrogen bond dissociation energies for naphthalene also were obtained, in this case via the application of a thermodynamic cycle. The final results are DeltaH(o)acid (1alpha) = 394.2+/-1.2 kcal mol(-1), DeltaH(o)acid (1beta) = 395.5+/-1.3 kcal mol(-1), EA(alpha) = 31.6+/-0.5 kcal mol(-1), EA(beta) = 31.6+/-0.5 kcal mol(-1), BDE(1alpha) = 112.2+/-1.3 kcal mol(-1) and BDE(1alpha) = 111.9+/-1.4 kcal mol(-1), and they are compared to benzene and phenyl radical as well as ab initio and density functional theory (B3LYP) calculations.


The 1.5-A structure of Chryseobacterium meningosepticum zinc beta-lactamase in complex with the inhibitor, D-captopril.
The crystal structure of the class-B beta-lactamase, BlaB, from the pathogenic bacterium, Chryseobacterium meningosepticum, in complex with the inhibitor, d-captopril, has been solved at 1.5-A resolution. The enzyme has the typical alphabeta/betaalpha metallo-beta-lactamase fold and the characteristic two metal binding sites of members of the subclass B1, in which two Zn2+ ions were identified. d-Captopril, a diastereoisomer of the commercial drug, captopril, acts as an inhibitor by displacing the catalytic hydroxyl ion required for antibiotic hydrolysis and intercalating its sulfhydryl group between the two Zn2+ ions. Interestingly, d-captopril is located on one side of the active site cleft. The x-ray structure of the complex of the closely related enzyme, IMP-1, with a mercaptocarboxylate inhibitor, which also contains a sulfhydryl group bound to the two Zn2+ ions, shows the ligand to be located on the opposite side of the active site cleft. A molecule generated by fusion of these two inhibitors would cover the entire cleft, suggesting an interesting approach to the design of highly specific inhibitors.


Recurrence of hepatocellular carcinoma following deceased donor liver transplantation: case series.
We aimed to study the clinical and pathological characteristics of liver transplant recipients with hepatocellular carcinoma recurrence. We reviewed the data for 26 patients who had tumor recurrence after deceased donor liver transplant for hepatocellular carcinoma at the Johns Hopkins Hospital from January 2005 to December 2015. In total, 88% of recipients were males. The mean age was 59 years. On explant, poor differentiation was detected in 43%, while 73% had microvascular invasion. Overall, 62% were diagnosed to be outside of Milan criteria. Out of these, 15% met the criteria for downstaging. Twenty (77%) patients had pre-transplant alpha fetoprotein levels ≥ 20 ng/mL. In 54% of patients, the location of hepatocellular carcinoma (HCC) recurrence was extrahepatic, followed by intrahepatic in 31% and both intra- and extrahepatic in 15%. The post-transplant tumor recurrence was diagnosed at a mean of 427 days (range 34-1502). Fifty percent of HCC recurrences were diagnosed within one year following liver transplant. Twenty (77%) patients received treatment for their recurrent HCC: external radiation (n = 10), surgical resections (n = 8; brain 4, spine 2, bone 1, and Whipple surgery 1), sorafenib (n = 7), locoregional therapy (n = 5). Overall, 24 out of 26 (92%) recipients died within four years after the transplant. HCC recurrence after liver transplant is infrequent. More than fifty percent of HCC recurrences following liver transplant are extrahepatic. Despite better recipient selection for liver transplant, the curative options are limited in recurrent cases and associated with extremely poor outcomes.


Dogs Rely On Visual Cues Rather Than On Effector-Specific Movement Representations to Predict Human Action Targets.
The ability to predict others' actions is one of the main pillars of social cognition. We investigated the processes underlying this ability by pitting motor representations of the observed movements against visual familiarity. In two pre-registered eye-tracking experiments, we measured the gaze arrival times of 16 dogs (Canis familiaris) who observed videos of a human or a conspecific executing the same goal-directed actions. On the first trial, when the human agent performed human-typical movements outside dogs' specific motor repertoire, dogs' gaze arrived at the target object anticipatorily (i.e., before the human touched the target object). When the agent was a conspecific, dogs' gaze arrived to the target object reactively (i.e., upon or after touch). When the human agent performed unusual movements more closely related to the dogs' motor possibilities (e.g., crawling instead of walking), dogs' gaze arrival times were intermediate between the other two conditions. In a replication experiment, with slightly different stimuli, dogs' looks to the target object were neither significantly predictive nor reactive, irrespective of the agent. However, when including looks at the target object that were not preceded by looks to the agents, on average dogs looked anticipatorily and sooner at the human agent's action target than at the conspecific's. Looking times and pupil size analyses suggest that the dogs' attention was captured more by the dog agent. These results suggest that visual familiarity with the observed action and saliency of the agent had a stronger influence on the dogs' looking behaviour than effector-specific movement representations in anticipating action targets.


Integrated 'omics analysis for the gut microbiota response to moxibustion in a rat model of chronic fatigue syndrome.
To observe the efficacy of moxibustion in the treatment of chronic fatigue syndrome (CFS) and explore the effects on gut microbiota and metabolic profiles. Forty-eight male Sprague-Dawley rats were randomly assigned to control group (Con), CFS model group (Mod, established by multiple chronic stress for 35 d), MoxA group (CFS model with moxibustion Shenque (CV8) and Guanyuan (CV4), 10 min/d, 28 d) and MoxB group (CFS model with moxibustion Zusanli (ST36), 10 min/d, 28 d). Open-field test (OFT) and Morris-water-maze test (MWMT) were determined for assessment the CFS model and the therapeutic effects of moxibustion.16S rRNA gene sequencing analysis based gut microbiota integrated untargeted liquid chromatograph-mass spectrometer (LC-MS) based fecal metabolomics were executed, as well as Spearman correlation analysis, was utilized to uncover the functional relevance between the potential metabolites and gut microbiota. The results of our behavioral tests showed that moxibustion improved the performance of CFS rats in the OFT and the MWMT. Microbiome profiling analysis revealed that the gut microbiomes of CFS rats were less diverse with altered composition, including increases in pro-inflammatory species (such as Proteobacteria) and decreases in anti-inflammatory species (such as Bacteroides, Lactobacillus, Ruminococcus, and Prevotella). Moxibustion partially normalized these changes in the gut microbiota. Furthermore, CFS was associated with metabolic disorders, which were effectively ameliorated by moxibustion. This was demonstrated by the normalization of 33 microbiota-related metabolites, including mannose (P = 0.001), aspartic acid (P = 0.009), alanine (P = 0.007), serine (P = 0.000), threonine (P = 0.027), methionine (P = 0.023), 5-hydroxytryptamine (P = 0.008), alpha-linolenic acid (P = 0.003), eicosapentaenoic acid (P = 0.006), hypoxanthine (P = 0.000), vitamin B6 (P = 0.000), cholic acid (P = 0.013), and taurocholate (P = 0.002). Correlation analysis showed a significant association between the perturbed fecal microbiota and metabolite levels, with a notable negative relationship between LCA and Bacteroides. In this study, we demonstrated that moxibustion has an antifatigue-like effect. The results from the 16S rRNA gene sequencing and metabolomics analysis suggest that the therapeutic effects of moxibustion on CFS are related to the regulation of gut microorganisms and their metabolites. The increase in Bacteroides and decrease in LCA may be key targets for the moxibustion treatment of CFS.


Cerebrospinal fluid inflammatory cytokines and chemokines in naturally occurring canine spinal cord injury.
Canine intervertebral disk herniation (IVDH) is a common, naturally occurring form of spinal cord injury (SCI) that is increasingly being used in pre-clinical evaluation of therapies. Although IVDH bears critical similarities to human SCI with respect to lesion morphology, imaging features, and post-SCI treatment, limited data are available concerning secondary injury mechanisms. Here, we characterized cerebrospinal fluid (CSF) cytokines, and chemokines in dogs with acute, surgically treated, thoracolumbar IVDH (n=39) and healthy control dogs (n=21) to investigate early inflammatory events after SCI. A bioplex system was used to measure interleukin (IL)-2, -6, -7, -8, -10, -15, and -18, granulocyte macrophage colony-stimulating factor (GM-CSF), interferon gamma (IFN-γ), keratinocyte chemoattractant (KC)-like protein, IFN-γ-inducible protein-10, monocyte chemotactic protein 1 (MCP-1), and tumor necrosis factor alpha. Cytokine and chemokine concentrations in the CSF of healthy and SCI dogs were compared and, in SCI dogs, were correlated to the duration of SCI, behavioral measures of injury severity at the time of sampling, and neurological outcome 42 days post-SCI as determined by a validated ordinal score. IL-8 concentration was significantly higher in SCI cases than healthy controls (p=0.0013) and was negatively correlated with the duration of SCI (p=0.042). CSF MCP-1 and KC-like protein were positively correlated with CSF microprotein concentration in dogs with SCI (p<0.0001 and p=0.004). CSF MCP-1 concentration was negatively associated with 42-day postinjury outcome (p<0.0001). Taken together, these data indicate that cytokines and chemokines present after SCI in humans and rodent models are associated with SCI pathogenesis in canine IVDH.


Control of vacuole permeability and protein degradation by the cell cycle arrest signal in Saccharomyces cerevisiae.
Saccharomyces cerevisiae responds to deperivation of nutrients by arresting cell division at the unbudded G1 stage. Cells situated outside of G1 at the time of deperivation complete the cell cycle before arresting. This prompted an investigation of the source of nutrients used by these cells to complete division and the mechanisms controlling their availability. We found a close correlation between accumulation of unbudded cells and loss of previously formed allophanate hydrolase activity after nutrient starvation. These losses were not specific to the allantoin, system since they have been observed for a number of other enzymes and also when cellular protein levels were monitored with [3H]leucine. Loss of hydrolase activity was also observed when protein synthesis was inhibited either by addition of inhibitors or loss of the prtl gene product. We found that onset of nutrient starvation brought about release of large quantities of arginine and allantoin normally sequestered in the cell vacuole. Treatment of a cells with alpha-factor resulted in both the release of allantoin and arginine from the cell vacuole and the onset of intracellular protein degradation. These effects were not observed when either alpha cells or a/alpha diploid strains were treated with alpha-factor. These data suggest that release of vacuolar constitutents and protein turnover may be regulated by the G1 arrest signal.


Transcriptome analysis unveils the mechanisms of lipid metabolism response to grayanotoxin I stress in Spodoptera litura.
Spodoptera litura (tobacco caterpillar, S. litura) is a pest of great economic importance due to being a polyphagous and world-distributed agricultural pest. However, agricultural practices involving chemical pesticides have caused resistance, resurgence, and residue problems, highlighting the need for new, environmentally friendly methods to control the spread of S. litura. This study aimed to investigate the gut poisoning of grayanotoxin I, an active compound found in Pieris japonica, on S. litura, and to explore the underlying mechanisms of these effects. S. litura was cultivated in a laboratory setting, and their survival rate, growth and development, and pupation time were recorded after grayanotoxin I treatment. RNA-Seq was utilized to screen for differentially expressed genes (DEGs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted to determine the functions of these DEGs. ELISA was employed to analyze the levels of lipase, 3-hydroxyacyl-CoA dehydrogenase (HOAD), and acetyl-CoA carboxylase (ACC). Hematoxylin and Eosin (H & E) staining was used to detect the development of the fat body. Grayanotoxin I treatment significantly suppressed the survival rate, growth and development, and pupation of S. litura. RNA-Seq analysis revealed 285 DEGs after grayanotoxin I exposure, with over 16 genes related to lipid metabolism. These 285 DEGs were enriched in the categories of cuticle development, larvae longevity, fat digestion and absorption. Grayanotoxin I treatment also inhibited the levels of FFA, lipase, and HOAD in the hemolymph of S. litura. The results of this study demonstrated that grayanotoxin I inhibited the growth and development of S. litura. The mechanisms might, at least partly, be related to the interference of lipid synthesis, lipolysis, and fat body development. These findings provide valuable insights into a new, environmentally-friendly plant-derived insecticide, grayanotoxin I, to control the spread of S. litura.


Neural bases of goal-directed implicit learning.
Several neuropsychological and neuroimaging studies have been performed to clarify the neural bases of implicit learning, but the question of which brain regions are involved in different forms of implicit learning, including goal-directed learning and habit learning, has not yet been resolved. The present study sought to clarify the mechanisms of goal-directed implicit learning by examining the sugar production factory (SPF) task in conjunction with functional magnetic resonance imaging (fMRI). Several brain regions were identified that contribute to learning in the SPF task. Significant learning-related decreases in brain activity were found in the right inferior parietal lobule (IPL), left superior frontal gyrus, right medial frontal gyrus, cerebellar vermis, and left inferior frontal gyrus, while significant learning-related increases in activity were observed in the right inferior frontal gyrus, left precenteral gyrus and, left precuneus. Among these regions, we speculate that the IPL and medial frontal gyrus may specifically be involved in the early stage of goal-directed implicit learning. We also attempted to investigate the role of the striatum, which has a significant role in habit learning, during learning of the SPF task. The results of ROI analysis showed no learning-related change in the activity of the striatum. Although some of the observed learning-related activations in this study have also been previously reported in neuroimaging studies of habit learning, the possibility that specific brain regions involved in goal-direct implicit learning cannot be excluded.


Structured approach to patients with memory difficulties in family practice.
To provide family physicians with a structured approach to patients presenting with memory difficulties. The approach is based on an accredited memory clinic training program developed by the Centre for Family Medicine Memory Clinic in partnership with the Ontario College of Family Physicians. Use of a structured clinical reasoning approach can assist physicians in achieving an accurate diagnosis in patients presenting with memory difficulties. Delirium, depression, and reversible causes need to be excluded, followed by differentiation among normal cognitive aging, mild cognitive impairment, and dementia. Obtaining collateral history and accurate functional assessment are critical. Common forms of dementia can be clinically differentiated by the order in which symptoms appear and by how cognitive deficits evolve over time. Typically, early signs of Alzheimer dementia involve impairment in episodic memory, whereas dementia involving predominantly vascular causes might present with early loss of executive function and relatively preserved episodic memory. Frontotemporal dementia and Lewy body spectrum disorders might have early loss of executive function and visuospatial function, as well as characteristic clinical features. A clinical reasoning approach can help physicians achieve early, accurate diagnoses that can guide appropriate management and improve care for patients with memory difficulties.


Shaped multi-cycle two-color laser field for generating an intense isolated XUV pulse toward 100 attoseconds.
The isolated attosecond pulse (IAP) generated from high-order harmonic (HH) radiation has been established as an important technique for the ultrafast optics over past decade. The applications of IAP in ultrafast processes can be greatly extended by further developing the high-intensity IAP. Here, we theoretically propose to shape a two-color field by performing peak amplitude-wavelength analysis. It is found that a 240-as IAP can be generated even without carrier envelop phase (CEP) stabilization using a 25 fs/800 nm fundamental field and a relative weak 25 fs/1330 nm control field, which enables us to markedly relax the requirements of the driving laser fields both in pulse duration and CEP control. On the other hand, if the CEPs of driving laser fields are stabilized, a 65-eV broadband continual harmonic, supporting a 81-as IAP, can be directly produced with the optimized intensity ratio of 0.866 and control wavelength of 1400 nm. Moreover, the propagation effect of two-color field on the macroscopic build-up of HH for generating a high-energy IAP is discussed. We found that the method of phase match still works for the efficient continuous harmonic generation as long as the ionization level and the pressure of gas medium are kept low enough. Since the phase-matched short IAP can be generated with our shaped two-color scheme in combination with a relaxed requirement of driving laser fields, the commercial available high-energy laser source with a loosely focused geometry is promising for scaling up the energy of IAP, showing the potential for the realization of IAP with high focused intensity toward 100 attoseconds.


Hepatitis B virus prevents excessive viral production via reduction of cell death-inducing DFF45-like effectors.
The relationship between hepatitis B virus (HBV) infection and lipid accumulation remains largely unknown. In this study, we investigated the effect of HBV propagation on lipid droplet growth in HBV-infected cells and HBV-producing cell lines, HepG2.2.15 and HBV-inducible Hep38.7-Tet. The amount of intracellular triglycerides was significantly reduced in HBV-infected and HBV-producing cells compared with HBV-lacking control cells. Electron and immunofluorescent microscopic analyses showed that the average size of a single lipid droplet (LD) was significantly less in the HBV-infected and HBV-producing cells than in the HBV-lacking control cells. Cell death-inducing DFF45-like effectors (CIDEs) B and C (CIDEB and CIDEC), which are involved in LD expansion for the improvement of lipid storage, were expressed at a significantly lower level in HBV-infected or HBV-producing cells than in HBV-lacking control cells, while CIDEA was not detected in those cells regardless of HBV production. The activity of the CIDEB and CIDEC gene promoters was impaired in HBV-infected or HBV-producing cells compared to HBV-lacking control cells, while CIDEs potentiated HBV core promoter activity. The amount of HNF4α, that can promote the transcription of CIDEB was significantly lower in HBV-producing cells than in HBV-lacking control cells. Knockout of CIDEB or CIDEC significantly reduced the amount of supernatant HBV DNA, intracellular viral RNA and nucleocapsid-associated viral DNA, while the expression of CIDEB or CIDEC recovered HBV production in CIDEB- or CIDEC-knockout cells. These results suggest that HBV regulates its own viral replication via CIDEB and CIDEC.


Biopsychosocial assessment criteria for functional chronic visceral pain: a pilot review of concept and practice.
Functional chronic visceral pain (FCVP) is one of the most common causes of morbidity in the general population. Pain perceived within the abdomen may occur due to a range of different mechanisms according to the organ and their afferent pathways. Advances in our understanding of the complexities of FCVP could lead to the exploitation of contemporary research in order to develop and utilize our understanding of neurobiological and psychobiological visceral mechanisms in a clinical setting. This progression, together with increasing amounts of epidemiological and gender based information concerning specific abdominal pain syndromes can allow us to develop assessment tools that go beyond disease only analysis and move toward a more comprehensive assessment model so that patients may have access to expert or multidisciplinary management sooner, rather than later. Based on current evidence, one must consider the main contributors to pain, whether it is nociceptive, neuropathic or psychosocial or as is common with FCVP, a combination of all three. This comprehensive assessment model should encompass not only systematic evaluation for reliable communication, but should also progress toward idiographic diagnosis relating to the uniqueness of the patient. This model should be practical in a multidisciplinary setting, taking into account the multi-faceted nature of this presentation.


DNA replicons for plant genome engineering.
Sequence-specific nucleases enable facile editing of higher eukaryotic genomic DNA; however, targeted modification of plant genomes remains challenging due to ineffective methods for delivering reagents for genome engineering to plant cells. Here, we use geminivirus-based replicons for transient expression of sequence-specific nucleases (zinc-finger nucleases, transcription activator-like effector nucleases, and the clustered, regularly interspaced, short palindromic repeat/Cas system) and delivery of DNA repair templates. In tobacco (Nicotiana tabacum), replicons based on the bean yellow dwarf virus enhanced gene targeting frequencies one to two orders of magnitude over conventional Agrobacterium tumefaciens T-DNA. In addition to the nuclease-mediated DNA double-strand breaks, gene targeting was promoted by replication of the repair template and pleiotropic activity of the geminivirus replication initiator proteins. We demonstrate the feasibility of using geminivirus replicons to generate plants with a desired DNA sequence modification. By adopting a general plant transformation method, plantlets with a desired DNA change were regenerated in <6 weeks. These results, in addition to the large host range of geminiviruses, advocate the use of replicons for plant genome engineering.


All CLEAR? Preparing for IT Downtime.
Health care information technology (IT) outages pose a threat to patient safety and patient care continuity. Organizations' downtime plans must be updated regularly and staff at the work area level should have experience with implementing IT outage operations through downtime drills. This article describes the study institution's IT Outage Toolkit, based on the acronym CLEAR, which guides the development of a downtime plan as well as design, execution, and assessment of work area downtime drills. Self-report and external audits of downtime drills help identify performance gaps and gaps in downtime plans.


Quantification of load-dependent changes in the collagen fiber architecture for the strut chordae tendineae-leaflet insertion of porcine atrioventricular heart valves.
Atrioventricular heart valves (AHVs) regulate the unidirectional flow of blood through the heart by opening and closing of the leaflets, which are supported in their functions by the chordae tendineae (CT). The leaflets and CT are primarily composed of collagen fibers that act as the load-bearing component of the tissue microstructures. At the CT-leaflet insertion, the collagen fiber architecture is complex, and has been of increasing focus in the previous literature. However, these previous studies have not been able to quantify the load-dependent changes in the tissue's collagen fiber orientations and alignments. In the present study, we address this gap in knowledge by quantifying the changes in the collagen fiber architecture of the mitral and tricuspid valve's strut CT-leaflet insertions in response to the applied loads by using a unique approach, which combines polarized spatial frequency domain imaging with uniaxial mechanical testing. Additionally, we characterized these microstructural changes across the same specimen without the need for tissue fixatives. We observed increases in the collagen fiber alignments in the CT-leaflet insertion with increased loading, as described through the degree of optical anisotropy. Furthermore, we used a leaflet-CT-papillary muscle entity method during uniaxial testing to quantify the chordae tendineae mechanics, including the derivation of the Ogden-type constitutive modeling parameters. The results from this study provide a valuable insight into the load-dependent behaviors of the strut CT-leaflet insertion, offering a research avenue to better understand the relationship between tissue mechanics and the microstructure, which will contribute to a deeper understanding of AHV biomechanics.


Eutrophication leads to the formation of a sulfide-rich deep-water layer in Lake Sevan, Armenia.
Lake Sevan is a meso-eutrophic water body, which was severely impacted by anthropogenic level decrease, pollution and eutrophication during the last century. Starting in the 1970s, these processes resulted in the formation of an oxygen-depleted hypolimnion during summer-autumn stratification of the lake. In this work, we demonstrate for the first time that eutrophication of the lake leads not only to the full depletion of oxygen and nitrate in the hypolimnion but as well to the presence of sulfate-reducing microorganisms and toxic hydrogen sulfide. Concentrations of hydrogen sulfide in the hypolimnion of Major and Minor Sevan in October were as high as 9 and 39 μM, respectively. In October 2019, 66 % of lake's bottom was covered by sulfidic waters, while the fraction of sulfidic water volume reached 19 %. Values of δ34S for hypolimnetic sulfide are lower by only 7-12 ‰ compared to epilimnetic sulfate, while δ33S values of sulfide are similar to the δ33S values of sulfate. These isotopic fingerprints are not consistent with microbial sulfate reduction as the sole source of hydrogen sulfide in the hypolimnion. We attribute the formation of a sulfidic deep-water layer to a combination of microbial sulfate reduction in the water column and diffusion of hydrogen sulfide from the sediments.


Measurement of amniotic fluid surfactant.
Measurement of the production of surfactant is the most direct means of prenatally assessing fetal pulmonary maturity. We review assays which have evolved for measuring surfactant, classifying them into two general categories: biochemical quantitation and biophysical measurements. Biochemical quantitation assays include the amniotic fluid lecithin/sphingomyelin ratio and quantitation of lecithin and other surfactant phospholipids. Biophysical measurements include measurement of surface-tension-lowering ability of extracts of amniotic fluid lipid and evaluation of surface-tension-related properties such as foam stability and microviscosity. Assays of surfactant are subject to certain pre-analysis sources of variation over which the analyst has no control, such as variability in total in vivo amniotic fluid volume, incomplete in vivo mixing of surfactant with amniotic fluid, and presence of contaminating blood or meconium. We also examine other factors such as centrifugation speed and time, and storage of the amniotic specimen before analysis. These factors can dramatically affect analyses, and must be carefully controlled by the analyst. In general, both biochemical and biophysical approaches to surfactant analysis are useful diagnostically. When properly performed, both approaches give results that correlate well, both with each other and with clinical outcome. Because "mature" and "immature" values overlap, none of the assays can completely eliminate false predictions, whether of fetal maturity or fetal immaturity.


Drug-eluting polymer implants in cancer therapy.
Drug-eluting polymer implants present a compelling parenteral route of administration for cancer chemotherapy. With potential for minimally invasive, image-guided placement and highly localized drug release, these delivery systems are playing an increasingly important role in cancer management. This is particularly true as the use of labile proteins and other bioactive molecules is likely to increase in the upcoming years. In this review, we present the current trends in the application of Pre-formed and in situ-forming systems as drug-eluting implants for cancer chemotherapy. We outline the clinically available options as well as up-and-coming technologies and their advantages and challenges. We also describe ongoing related innovations with image-guided drug delivery, mathematical modeling of implanted delivery systems and implanted drug delivery in combination with other therapies. Whether used alone or combined with other minimally invasive procedures, drug-eluting polymeric implants will play a significant role in the future of cancer management.


First molecular characterization of Cryptosporidium in Yemen.
Cryptosporidium is a protozoan parasite of humans and animals and has a worldwide distribution. The parasite has a unique epidemiology in Middle Eastern countries where the IId subtype family of Cryptosporidium parvum dominates. However, there has been no information on Cryptosporidium species in Yemen. Thus, this study was conducted in Yemen to examine the distribution of Cryptosporidium species and subtype families. Fecal samples were collected from 335 patients who attended hospitals in Sana'a city. Cryptosporidium species were determined by PCR and sequence analysis of the 18 s rRNA gene. Cryptosporidium parvum and C. hominis subtypes were identified based on sequence analysis of the 60 kDa glycoprotein (gp60) gene. Out of 335 samples, 33 (9.9%) were positive for Cryptosporidium. Of them, 97% were identified as C. parvum whilst 1 case (3%) was caused by C. hominis. All 7 C. parvum isolates subtyped belonged to the IIaA15G2R1 subtype. The common occurrence of the zoonotic IIa subtype family of C. parvum highlights the potential occurrence of zoonotic transmission of cryptosporidiosis in Yemen. However, this postulation needs confirmation with future molecular epidemiological studies of cryptosporidiosis in both humans and animals in Yemen.


Megalencephalic Leukoencephalopathy: Insights Into Pathophysiology and Perspectives for Therapy.
Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is a rare genetic disorder belonging to the group of vacuolating leukodystrophies. It is characterized by megalencephaly, loss of motor functions, epilepsy, and mild mental decline. In brain biopsies of MLC patients, vacuoles were observed in myelin and in astrocytes surrounding blood vessels. It is mainly caused by recessive mutations in MLC1 and HEPACAM (also called GLIALCAM) genes. These disease variants are called MLC1 and MLC2A with both types of patients sharing the same clinical phenotype. Besides, dominant mutations in HEPACAM were also identified in a subtype of MLC patients (MLC2B) with a remitting phenotype. MLC1 and GlialCAM proteins form a complex mainly expressed in brain astrocytes at the gliovascular interface and in Bergmann glia at the cerebellum. Both proteins regulate several ion channels and transporters involved in the control of ion and water fluxes in glial cells, either directly influencing their location and function, or indirectly regulating associated signal transduction pathways. However, the MLC1/GLIALCAM complex function and the related pathological mechanisms leading to MLC are still unknown. It has been hypothesized that, in MLC, the role of glial cells in brain ion homeostasis is altered in both physiological and inflammatory conditions. There is no therapy for MLC patients, only supportive treatment. As MLC2B patients show an MLC reversible phenotype, we speculated that the phenotype of MLC1 and MLC2A patients could also be mitigated by the re-introduction of the correct gene even at later stages. To prove this hypothesis, we injected in the cerebellar subarachnoid space of Mlc1 knockout mice an adeno-associated virus (AAV) coding for human MLC1 under the control of the glial-fibrillary acidic protein promoter. MLC1 expression in the cerebellum extremely reduced myelin vacuolation at all ages in a dose-dependent manner. This study could be considered as the first preclinical approach for MLC. We also suggest other potential therapeutic strategies in this review.


Can physician assistants be effective in the UK?
The National Health Sevice (NHS) faces a serious shortage of medical staff. One solution is to introduce US-style physician assistants (PAs) who train for around two years following previous clinical work or a first degree, and perform duties similar to junior doctors. This paper reviews the history and role of PAs, the quality of their work and their likely impact in the UK. A variety of sources were searched to identify suitable studies. The use of PAs in the UK appears to be an acceptable model that could eventually reduce the current skill shortage and provide high quality patient care. Twelve US-sourced PAs currently work in Sandwell, West Midlands. A recent report suggests they have made a substantial contribution to primary care and have improved patient access. For PAs to be successful in the UK, they must be highly regarded practitioners. High quality educational courses must be established to ensure their credibility.


Practical Guidance for Clinical Microbiology Laboratories: Viruses Causing Acute Respiratory Tract Infections.
Respiratory viral infections are associated with a wide range of acute syndromes and infectious disease processes in children and adults worldwide. Many viruses are implicated in these infections, and these viruses are spread largely via respiratory means between humans but also occasionally from animals to humans. This article is an American Society for Microbiology (ASM)-sponsored Practical Guidance for Clinical Microbiology (PGCM) document identifying best practices for diagnosis and characterization of viruses that cause acute respiratory infections and replaces the most recent prior version of the ASM-sponsored Cumitech 21 document, Laboratory Diagnosis of Viral Respiratory Disease, published in 1986. The scope of the original document was quite broad, with an emphasis on clinical diagnosis of a wide variety of infectious agents and laboratory focus on antigen detection and viral culture. The new PGCM document is designed to be used by laboratorians in a wide variety of diagnostic and public health microbiology/virology laboratory settings worldwide. The article provides guidance to a rapidly changing field of diagnostics and outlines the epidemiology and clinical impact of acute respiratory viral infections, including preferred methods of specimen collection and current methods for diagnosis and characterization of viral pathogens causing acute respiratory tract infections. Compared to the case in 1986, molecular techniques are now the preferred diagnostic approaches for the detection of acute respiratory viruses, and they allow for automation, high-throughput workflows, and near-patient testing. These changes require quality assurance programs to prevent laboratory contamination as well as strong preanalytical screening approaches to utilize laboratory resources appropriately. Appropriate guidance from laboratorians to stakeholders will allow for appropriate specimen collection, as well as correct test ordering that will quickly identify highly transmissible emerging pathogens.


Oral transit time: a critical review of the literature.
Oral transit time is one of the parameters observed during the clinical assessment of the swallowing function. The importance of this parameter is due to its impact on the total duration of a meal, whose consequence can be an unfavorable nutritional prognostic. To document scientific papers that measure oral transit time in healthy subjects. The review followed the steps proposed by the Cochrane Handbook. The search was done via the PubMed database through the use of descriptors related to the oral phase of swallowing, as well as to types of food consistency. The articles on the theme had different definitions for oral transit time, as well as heterogeneity of tested volumes, age and gender of the participants. The times found varied from 0.35 s to 1.54 s for liquids, from 0.39 s to 1.05 s for pasty foods and from 1 s to 12.8 s for solid foods. Also, regardless of volume or consistency, oral transit time in elderly people is significantly longer than in adults. There's no consensus in the literature about oral transit time in healthy subjects. However, this parameter should be valued during the assessment of the swallowing function due to its negative impact on the dynamics of swallowing, which can cause high energy expenditure during feeding.


IL-4 type 1 receptor signaling up-regulates KCNN4 expression, and increases the KCa3.1 current and its contribution to migration of alternative-activated microglia.
The Ca(2+)-activated K(+) channel, KCa3.1 (KCNN4/IK1/SK4), contributes to "classical," pro-inflammatory activation of microglia, and KCa3.1 blockers have improved the outcome in several rodent models of CNS damage. For instance, blocking KCa3.1 with TRAM-34 rescued retinal ganglion neurons after optic nerve damage in vivo and, reduced p38 MAP kinase activation, production of reactive oxygen and nitrogen species, and neurotoxicity by microglia in vitro. In pursuing the therapeutic potential of KCa3.1 blockers, it is crucial to assess KCa3.1 contributions to other microglial functions and activation states, especially the IL-4-induced "alternative" activation state that can counteract pro-inflammatory states. We recently found that IL-4 increases microglia migration - a crucial function in the healthy and damaged CNS - and that KCa3.1 contributes to P2Y2 receptor-stimulated migration. Here, we discovered that KCa3.1 is greatly increased in alternative-activated rat microglia and then contributes to an enhanced migratory capacity. IL-4 up-regulated KCNN4 mRNA (by 6 h) and greatly increased the KCa3.1 current by 1 day, and this required de novo protein synthesis. The increase in current was sustained for at least 6 days. IL-4 increased microglial migration and this was reversed by blocking KCa3.1 with TRAM-34. A panel of inhibitors of signal-transduction mediators was used to analyze contributions of IL-4-related signaling pathways. Induction of KCNN4 mRNA and KCa3.1 current was mediated specifically through IL-4 binding to the type I receptor and, surprisingly, it required JAK3, Ras/MEK/ERK signaling and the transcription factor, activator protein-1, rather than JAK2, STAT6, or phosphatidylinositol 3-kinase.The same receptor subtype and pathway were required for the enhanced KCa3.1-dependent migration. In providing the first direct signaling link between an IL-4 receptor, expression and roles of an ion channel, this study also highlights the potential importance of KCa3.1 in alternative-activated microglia.


Purulent flexor tenosynovitis: factors influencing the functional outcome.
This retrospective study aimed to evaluate the factors that influence the final outcome of treatment of purulent flexor tenosynovitis, viz. delay in treatment, severity of the condition, the infecting pathogen and the method of treatment. Of 41 patients with this condition treated by drainage and irrigation through two small incisions (16) and wide incision (25), 16 were treated after a delay. Continuous postoperative irrigation was applied in 26 patients. Re-operation was necessary in 11 patients. In most cases, Staphylococcus aureus was detected. The results were excellent in 32 cases and the mean Disabilities of the Arm, Shoulder and Hand score was 8.1. Irrigation through small incisions and continuous postoperative irrigation decreased the probability of an unfavourable outcome. Delayed treatment and infections with specific pathogens led to a worse outcome. Early diagnosis of purulent flexor tenosynovitis followed by drainage through small incisions and continuous postoperative irrigation appear, from this retrospective review, to lead to the best functional outcome.


The influence of a move to program management on physical therapist practice.
The purpose of this qualitative study was to examine how a move to program management (PM) from a traditional department structure affected the professional practice of physical therapists in a large Canadian teaching hospital. Twenty-five physical therapists participated in 1 of 5 focus groups, and 4 physical therapists participated in individual interviews. Focus groups and structured interviews were conducted by an experienced facilitator who was not a physical therapist. All focus groups and interviews were audiotaped and transcribed. Using an open-coding technique, 2 investigators undertook line-by-line analysis of each transcript to identify and code specific events related to the physical therapists' experiences. The investigators reached a consensus on all coding categories and then identified themes. Seven themes that addressed issues of affect (a sense of loss, low morale, and positive coping), professional practice (loss of professional development activities, professional advantages, the assuming of multiple roles), and patient care were identified. Physical therapists who were deployed from a department to a program described both positive and negative effects of the move to PM on their practice. There were reported influences on their personal affect, professional practice, and patient care. Staff and physical therapy administrators need to be aware of potential implications of an organization's move to PM on the professional practice of frontline staff.


Suicide in india: Part perceptions, partial insights, and inadequate solutions.
Suicide is a complex phenomenon, often linked to environment. Despite the identification of many social, cultural, economic and political correlates and risk factors, psychiatry continues to argue for curative solutions based on the reductionistic biomedical model, rather than support public health measures to manage the larger sociocultural, economic and political context. While psychiatry and curative medicine help many people, survival of the human body is best explained by the materialist explanation that locates the variation in health and longevity to tangible resources. There is no single, simple or straightforward solution to reducing population suicide rates; specific mental health interventions are unlikely to impact secular trends in the rates of suicide.


The reporting of funding in health policy and systems research: a cross-sectional study.
Major research-reporting statements, such as PRISMA and CONSORT, require authors to provide information about funding. The objectives of this study were (1) to assess the reporting of funding in health policy and systems research (HPSR) papers and (2) to assess the funding reporting policies of journals publishing on HPSR. We conducted two cross-sectional surveys for papers published in 2016 addressing HPSR (both primary studies and systematic reviews) and for journals publishing on HPSR (both journals under the 'Health Policy and Services' (HPS) category in the Web of Science, and non-HPS journals that published on HPSR). Teams of two reviewers selected studies and abstracted data in duplicate and independently. We conducted descriptive analyses and a regression analysis to investigate the association between reporting of funding by papers and the journal's characteristics. We included 400 studies (200 systematic reviews and 200 primary studies) that were published in 198 journals. Approximately one-third (31%) of HPSR papers did not report on funding. Of those that did, only 11% reported on the role of funders (15% of systematic reviews and 7% of primary studies). Of the 198 journals publishing on HPSR, 89% required reporting of the source of funding. Of those that did, about one-third (34%) required reporting of the role of funders. Journals classified under the HPS category (n = 72) were less likely than non-HPS journals that published HPSR studies (n = 142) to require information on the role of funders (15% vs. 32%). We did not find any of the journals' characteristics to be associated with the reporting of funding by papers. Despite the majority of journals publishing on HPSR requiring the reporting of funding, approximately one-third of HPSR papers did not report on the funding source. Moreover, few journals publishing on HPSR required the reporting of the role of funders, and few HPSR papers reported on that role.


125I-lysergic acid diethylamide binds to a novel serotonergic site on rat choroid plexus epithelial cells.
125I-Lysergic acid diethylamide (125I-LSD) binds with high affinity to serotonergic sites on rat choroid plexus. These sites were localized to choroid plexus epithelial cells by use of a novel high resolution stripping film technique for light microscopic autoradiography. In membrane preparations from rat choroid plexus, the serotonergic site density was 3100 fmol/mg of protein, which is 10-fold higher than the density of any other serotonergic site in brain homogenates. The choroid plexus site exhibits a novel pharmacology that does not match the properties of 5-hydroxytryptamine-1a (5-HT1a), 5-HT1b, or 5-HT2 serotonergic sites. 125I-LSD binding to the choroid plexus site is potently inhibited by mianserin, serotonin, and (+)-LSD. Other serotonergic, dopaminergic, and adrenergic agonists and antagonists exhibit moderate to weak affinities for this site. The rat choroid plexus 125I-LSD binding site appears to represent a new type of serotonergic site which is located on non-neuronal cells in this tissue.


Greek Sage Exhibits Neuroprotective Activity against Amyloid Beta-Induced Toxicity.
Alzheimer's disease (AD) is the most common neurodegenerative disease, affecting the elderly at a high incidence. AD is of unknown etiology and currently, no cure is available. Present medication is restricted to treating symptoms; thus, a need exists for the development of effective remedies. Medicinal plants constitute a large pool, from which active compounds of great pharmaceutical potential can be derived. Various Salvia spp. are considered as neuroprotective, and here, the ability of Salvia fruticosa (SF) to protect against toxic effects induced in an AD cell model was partly assessed. Two of AD's characteristic hallmarks are the presence of elevated oxidative stress levels and the cytotoxic aggregation of amyloid beta (Aβ) peptides. Thus, we obtained SF extracts in three different solvents of increasing polarity, consecutively, to evaluate (a) their antioxidant capacity with the employment of the free radical scavenging assay (DPPH•), of the ferric reducing ability of plasma assay (FRAP), and of the cellular reactive oxygen species assay (DCFDA) and (b) their neuroprotective properties against Aβ 25-35-induced cell death with the use of an MTT assay. All three SF extracts showed a considerable antioxidant capacity, with the methanol (SFM) extract being the strongest. The results of the total phenolic and flavonoid contents (TPC and TFC) of the extracts and of the FRAP and the DCFDA assays showed a similar pattern. In addition, and most importantly, the dichloromethane (SFD) and the petroleum ether (SFP) extracts had an effect on Aβ toxicity, exhibiting a significant neuroprotective potential. To our knowledge, this is the first report of SF extracts demonstrating neuroprotective potential against Aβ toxicity. In combination with their antioxidant capacity, SF extracts may be beneficial in combating AD and other neurodegenerative diseases.


Pharmacologically based dosing of etoposide: a means of safely increasing dose intensity.
We have previously demonstrated that individualized dosing of etoposide (VP16) by 72-hour infusion is feasible and that the extent of leukopenia is a function of plasma concentration, pretreatment WBC (WBCp), albumin (ALB), performance status (PS), and bone marrow function (based on transfusion requirements). In the current study, 45 patients were randomized between a fixed dose of VP16 (125 mg/m2/d) versus individualized dosing to a target WBC nadir (WBCN) of 1,700/microL. The total dose was increased by an average of 22% in the latter patients (459 +/- 130 mg/m2 v 375 mg/m2, P = .002). This was associated with a decrease in both the mean WBCN (1,510 +/- 950 v 2,500 +/- 1,420/microL, P = .013) and in the variability of the WBCN (P = .039). The VP16 clearance (mL/min) was not correlated with body surface area. Partial responses were observed in one patient each with hepatoma and non-Hodgkin's lymphoma. We conclude that pharmacologically based dosing may be a means of increasing dose intensity without increasing the incidence of life-threatening toxicity due to a decrease in variability around a target WBC.


Profiles of Chemical Effects on Cells (pCEC): a toxicogenomics database with a toxicoinformatics system for risk evaluation and toxicity prediction of environmental chemicals.
Profiles of Chemical Effects on Cells (pCEC) is a toxicogenomics database with a system of classifying chemicals that have effects on human health. This database stores and handles gene expression profiling information and categories of toxicity data. Chemicals are classified according to the specific tissues and cells they affect, the gene expression changes they induce, their toxicity and biological functions in this database system. The pCEC system also analyzes relationships between chemicals and the genes they affect in specific tissues and cells. The reason why we developed pCEC is to support decision-making within the context of environmental regulation. Especially, exposure to environmental chemicals during fetal and newborn development may result in a predisposition to various disorders such as cancer, learning disabilities and allergies later in life. The identification and prediction of hazardous chemicals using limited information are important issues in human health risk management. Therefore, various toxicity information including lethal dose 50 (LD50), toxicity pathways and pathological data were loaded into pCEC. pCEC is also a facility for query, analysis and prediction of unknown toxicochemical reaction pathways and biomarkers which are based on toxicoinformatical data mining approaches. This database is available online at http://project.nies.go.jp/eCA/cgi-bin/index.cgi. The current version of the database has information on the hepatotoxicity, reproductive toxicity and embryotoxicity of chemicals.


State Variations in Public Payer Reimbursement for Common Plastic Surgery Procedures.
Existing data suggest decreased willingness of plastic surgeons to participate in Medicare and Medicaid. Significant disparities exist in Medicare and Medicaid reimbursement for various general surgical procedures. The aims of this study were to investigate variations in Medicare and Medicaid reimbursement across the nation for common plastic surgery procedures. Medicare and Medicaid reimbursement data for 2017 were obtained by means of the Centers for Medicare & Medicaid Services and publicly available fee schedules from each state, respectively, for eight common plastic surgery procedures. The difference in Medicare and Medicaid reimbursement was calculated across all states. The difference in value ascribed to each procedure was determined by comparing the payment from each payer to the work relative value units. Medicaid reimbursement rates were significantly lower for the selected procedures, with a median national discount of -25 percent ($16.09 per work relative value unit) compared to Medicare. There were higher median rates of reimbursement per work relative value unit by Medicaid in only five states when compared to Medicare. Significant variations of more than 15 percent in the Medicaid-to-Medicare reimbursement ratios between our selected procedures were identified in 28 states. Variations exist between Medicare and Medicaid reimbursement for common plastic surgery procedures. The within-state variations in Medicaid reimbursement are likely reflective of important yet nontransparent differences in determining Medicaid reimbursement. These variations likely affect access to care for underserved populations. Professional societies should continue to convey the value of these important procedures and raise awareness regarding disparities in access to care.


Identification of C-type natriuretic peptide in heart of spiny dogfish shark (Squalus acanthias).
An unidentified atrial natriuretic peptide (ANP)-like substance is the principal hormone regulating NaCl secretion in the shark rectal gland, an epithelial model tissue for hormone-sensitive secondary active chloride transport. Antibodies to mammalian ANP do not recognize the prohormone of marine species. The polymerase chain reaction (PCR) was used to isolate a partial cDNA encoding the shark heart natriuretic peptide. Using this partial sequence as a probe, the full-length clone [882 base pairs (bp)] was obtained from a shark heart cDNA library. Amino acids 119-135 are similar to the recently identified peptide sequences of porcine C-type natriuretic peptide (CNP) and killifish brain natriuretic peptide isolated from the brain of these species. Mature shark heart CNP terminates at the second cysteine residue and lacks a COOH-terminal extension, in contrast to cardiac ANP-like peptides of all other species. The primary amino acid sequence of the shark heart prepro-CNP is distinctly different from all other cardiac natriuretic peptides. Amplification of genomic DNA spanning the coding region produced a 1.5-kb product, indicating the presence of at least one intron. Sequencing confirmed the presence of two exons of 90 and 315 bp, separated by a 1.1-kb intron. This is the first report of a cDNA encoding in nonneuronal tissue. Elasmobranch CNP may represent a primordial form of ANP-like peptides that evolved as an adaptation to environmental osmoregulatory stress.


Magnesium: potential roles in neurovascular disease.
Magnesium therapy has been studied extensively in pre-clinical and clinical trials in multiple organ systems. Cerebrovascular diseases may benefit from its neuroprotective properties. This review summarizes current studies of magnesium in a wide range of neurovascular diseases. We searched relevant terms in the National Library of Medicine PubMed database and selected research including basic science, translational reports, meta-analyses, and clinical studies. Studies examining magnesium administration in ischemic stroke have failed to show any benefit in clinical outcome. Data on magnesium for intracerebral hemorrhage (ICH) are limited. Preliminary investigations in subarachnoid hemorrhage (SAH) were promising, but definitive studies did not reveal differences in clinical outcome between magnesium and placebo-treated groups. Studies examining magnesium administration in global ischemia following cardiac arrest suggest a trend toward improved clinical outcome. The strongest evidence for clinically relevant neuroprotection following magnesium administration derives from studies of pre-term infants and patients undergoing cardiac bypass and carotid endarterectomy procedures. Magnesium was found to have an excellent safety profile across all investigations. Magnesium is easy to administer and possesses a favorable safety profile. Its utility as a neuroprotectant in cardiac surgery, carotid endarterectomy, and pre-term infant hypoxia remain promising. Value as a therapeutic agent in ischemic stroke, ICH, and SAH is unclear and appears to be limited by late administration. Ongoing clinical trials assessing magnesium administration in the first hours following symptom onset may help clarify the role of magnesium therapy in these disease processes.