Insights into Chemical Structure-Based Modeling for New Sweetener Discovery.
The search for novel, natural, high-sweetness, low-calorie sweeteners remains open and challenging. In the present study, the structure-based machine learning modeling and sweetness recognition mechanism were investigated to assist this process. It was found that whether or not a compound was sweet was closely related to molecular connectivity and composition (the number of hydrogen bond acceptors and donors), tpsaEfficiency, structural complexity, and shape (nAtomP and Fsp3). While the relative sweetness of sweet compounds was more determined by the molecular properties (tpsaEfficiency and Log P), structural complexity and composition (nAtomP and ATSm 1). The built machine learning models exhibited very good performance for classifying the sweet/non-sweet compounds and predicting the relative sweetness of the compounds. Moreover, a specific binding pocket was found for sweet compounds, and the sweet compounds mainly interacted with the VFT domain of the T1R2-T1R3 through hydrogen bonds. In addition, the results indicated that among the sweet compounds, those that were sweeter bound to the VFT domain stronger than those that had low sweetness. This study provides very useful information for developing new sweeteners.


Suppression Effect of Astaxanthin on Osteoclast Formation In Vitro and Bone Loss In Vivo.
Osteoporosis is characterized by a reduction of the bone mineral density (BMD) and microarchitectural deterioration of the bone, which lead to bone fragility and susceptibility to fracture. Astaxanthin (AST) has a variety of biological activities, such as a protective effect against asthma or neuroinflammation, antioxidant effect, and decrease of the osteoclast number in the right mandibles in the periodontitis model. Although treatment with AST is known to have an effect on inflammation, no studies on the effect of AST exposure on bone loss have been performed. Thus, in the present study, we examined the antiosteoporotic effect of AST on bone mass in ovariectomized (OVX) mice and its possible mechanism of action. The administration of AST (5, 10 mg/kg) for 6 weeks suppressed the enhancement of serum calcium, inorganic phosphorus, alkaline phosphatase, total cholesterol, and tartrate-resistant acid phosphatase (TRAP) activity. The bone mineral density (BMD) and bone microarchitecture of the trabecular bone in the tibia and femur were recovered by AST exposure. Moreover, in the in vitro experiment, we demonstrated that AST inhibits osteoclast formation through the expression of the nuclear factor of activated T cells (NFAT) c1, dendritic cell-specific transmembrane protein (DC-STAMP), TRAP, and cathepsin K without any cytotoxic effects on bone marrow-derived macrophages (BMMs). Therefore, we suggest that AST may have therapeutic potential for the treatment of postmenopausal osteoporosis.


Weight loss with interferon and ribavirin therapy in chronic hepatitis C patients.
Recommended treatment of chronic Hepatitis C virus (HCV) infection is interferon and ribavirin. Most patients develop side effects during treatment. Weight loss is one of the side effects of interferon therapy. Present study was conducted at Pakistan Medical and Research Centre (PMRC), Jinnah Postgraduate Medical Centre (JPMC), Karachi from January 1998 - December 2010, to find out the frequency, extent of weight loss, its recovery after interferon therapy and the impact of weight loss on treatment response. Body weight was recorded on regular outpatient visits i.e initially, then at 1, 3 and 6 months of treatment and six months post treatment. Out of 260 patients, 176 (67.7%) observed statistically significant weight loss (p<0.05), on completion of therapy as compared to initial visit. Weight loss during interferon therapy was found to be independent of age, gender, treatment outcome, serum albumin, haemoglobin, white blood cell (WBC) and platelet count. Body weight returned to baseline within six months after completion of treatment in most cases.


[Relation between Der p2 and Der f2 antigen levels in house dust and the degree of sensitization in asthmatic patients].
The continuous exposure to high levels of environmental allergens, as dust mite, gets worse the reactivity of the airways as mentioned in previous studies. In other international studies, it has been determined the levels of house dust mite antigen needed to sensitize asthmatic patients, but few studies in Mexico have measured the concentration of house dust mite antigen before the initial treatment and correlated the sensitization to it with the levels of total IgE and the skin test. To determine the minimum concentration of house dust mite antigen required to sensitize patients with asthma at the Distrito Federal and if it exists a correlation between the different concentration of the antigen, the results of the skin test and the total IgE. Thirty patients with asthma and positive skin test to Dermatophagoides pteronyssinus took part in the study. We made a home visit to measure the concentration of Der p2 and Der f2 in house dust and we scheduled a visit at our hospital to measure the IgE total level. We did a statistical analysis with the chi square test and the correlation coefficient of Pearson and we found that the lower level needed to sensitize was 0.2 micrograms of antigen per gram of house dust. It did not exist a correlation between the total IgE levels and the concentration of the mite antigen. To a higher level of the house dust mite antigen we found more sensitization to it, as shown by the skin test but without direct relation with the total IgE levels in patients with asthma.


Dietary fat type and level influence adiposity development in obese but not lean Zucker rats.
The development of obesity is influenced by a variety of factors including genetics and dietary fat type and level. To examine the interaction between these factors, male lean and obese Zucker rats (5 weeks initial age) were fed either a low-fat (15% calories) or one of two high-fat diets (65% calories; predominant fat source of either soybean oil or palm olein) for 8 weeks. Body weight, food intake, indirect calorimetry, and body composition determinations were performed. As expected, food intake, body weight, feed efficiency, oxygen consumption, heat production and carcass lipid were all significantly higher in obese compared to lean rats. Dietary fat level and/or type influenced body weight gain, oxygen consumption, heat production, energy balance, and carcass weight and lipid content in the obese but not in the lean Zucker rats. Oxygen consumption and carcass weight were increased approximately 25% and 10% respectively in obese rats fed either of the two high-fat diets as compared to those fed the low-fat diet. The type of fat fed in the high-fat diets also influenced body weight gain, heat production, energy balance, and carcass lipid content of the obese rats. Body weight gain and carcass lipid content were increased (16%-17%; P < 0.005) in obese rats fed the high-fat palm olein diet as compared to those fed the low-fat diet. These parameters were not increased in obese rats fed the high-fat soybean oil diet. In contrast, indirect calorimetry measurements indicated a moderate increase in heat production (Kcal/effective body mass/day; 14.5%) and decrease in energy balance (44.8%) in the obese rats fed the high-fat soybean oil diet as compared to those fed the low-fat diet. Energy expenditure and lipid accumulation were negligibly influenced by dietary fat level or type in the lean Zucker rats. The differential response of the lean and obese Zucker rats to this short-term dietary manipulation demonstrate that genetic background can influence an individual's response to dietary fat type and level. The genetically obese Zucker rat appears to be a good model for further studies of high-fat diet-induced obesity.


Mapping the frequency of malnutrition in patients with head and neck cancer using the GLIM Criteria for the Diagnosis of Malnutrition.
Patients with head and neck cancer are defined as high-risk patients for malnutrition, but the inconsistent practice of diagnosing malnutrition is a barrier in comparing studies and in assessing patients in clinical practice. The aim of the study was to describe the frequency of malnutrition over time in patients treated for head and neck cancer using the GLIM Criteria for the Diagnosis of Malnutrition. Data from a prospective observational study on patients with head and neck cancer were used (n = 210). Patients were assessed for malnutrition using the combination of one phenotypic and one etiologic criterion. The following phenotypic criteria for malnutrition were used: body weight loss (either >5% within the past six months or >10% beyond six months), body mass index (<20 kg/m2 if <70 years or <22 kg/m2 if ≥70 years), and fat free mass index measured by bioelectrical impedance analysis (<17 FFM/m2 for males and <15 FFM/m2 for females). The following etiologic criteria for malnutrition were used: reduced food intake (partial or no food intake with the need for artificial nutrition) and C-reactive protein (>5 mg/L). For all eight possible combinations of GLIM, the frequency of malnutrition reached its highest point at seven weeks after the start of treatment. A total of 32% of the patients fulfilled the criteria for malnutrition using weight loss >5% within the past six months (phenotypic) in combination with C-reactive protein >5 mg/L (etiologic). GLIM Criteria for the Diagnosis of Malnutrition can be used to assess malnutrition in patients with head and neck cancer during treatment. Using the same criteria to define malnutrition in nutritional research will make it possible to produce multiple lines of evidence on malnutrition in head and neck cancer and its adverse effects on treatment, prognosis, and survival.


Calling All Curators: A Novel Approach to Individualized Interactive Instruction.
With the increasing influence of the "Free Open Access Medical Education" (FOAM or FOAMed) movement, it is critical that medical educators be engaged with FOAM in order to better inform and direct their learners, who likely regularly consume these materials. In 2012, the Accreditation Council for Graduate Medical Education (ACGME)/Residency Review Committee (RRC) began to permit 20% of emergency medicine (EM) residents' didactics hours to be earned outside of weekly conference, as "Individualized Interactive Instruction" (III) credits.1 We describe a digital course in EM, "Asynchrony," as an approach to FOAM to meet these III standards. Asynchrony is geared toward EM residents using FOAM and other online learning tools, curated by faculty into narrative, topic-specific educational modules. Each module requires residents to complete a topic assignment, participate in a discussion board, and pass a quiz to earn ACGME-approved III didactic credit; all of this is tracked and filed in an online learning management system.


Whose cancer? Visualising the distribution of mentions to cancer sites on instagram.
This article presents a quantitative analysis of mentions to cancer on Instagram. Using thousands of images with cancer-related hashtags, we build several visualisations to capture their distribution. Source images are clustered by their visual traits and by the incidence, prevalence, and mortality of the cancer site they refer to. Our goal is three-fold: to provide a quantitative basis for future research on the representation of cancer online; to offer an interpretation of the sources of the imbalanced representation of the different cancer sites; and to motivate a debate on how that representation may affect patients and families.


Molecular modeling of eluted peptides from DQ6 molecules (DQB1*0602 and DQB1*0604) negatively and positively associated with type 1 diabetes.
Insulin-dependent diabetes mellitus (IDDM) is positively associated with HLA-DQ8, DQ2, and DQ6 (B*0604) and negatively with DQ6 (B*0602). The mechanisms by which the DQ molecules control the development of IDDM is not known. DQ6 (B*0602) and DQ6 (B*0604) molecules share the same DQalpha chain but differ in the beta chain by six residues at positions 9, 30, 57, 70, 86, and 87. The aim of the study was to sequence the peptides eluted from both DQ6 molecules and to determine the binding motifs and construct peptides for docking them into the DQ6 peptide binding groove by molecular modeling. EBV transformed B cell line homozygous for DQ6 and hybridoma cell line secreting DQ6 specific antibody were grown in large-scale culture. The DQ6 molecules were precipitated with solid-phase bound antibodies specific for DQ6. The dissociation of peptides from MHC was done with ultrafiltration and separation of peptides by reversed-phase HPLC, using Edman degradation. A special application of Edman degradation is pool sequencing. This approach allowed us to determine common characteristics of all peptides associated with a given MHC molecule. The precipitation of DQ6 molecules and the peptide elution were done successfully. The sequencing of the peptides from DQ6 (B*0602) identified three fractions: (1) IINEPTAAAIAYGLD (Bovine HSP70), (2) IINEPTAAAIAGLDR (Human HSP70), and (3) NPRDAKACVVHGSDLK (Na+/K+ ATPase). Peptide eluted from DQ6 (B*0604) had a sequence ADLFRGTLDPVEK with sequence homology to HSP70 (residues 307-319). We were able to predict the motifs for DQ6 from the ligands eluted. We used molecular modeling as a tool to identify plausible binding motifs for peptides. Our studies show that peptide ADLFRGTLDPVEK and NPRDAKACVVHGSDLK fit well in the respective DQ6 binding grooves. These predicted motifs should then be useful for screening of autoantigens associated with diabetes and identifying the epitopes that are likely to interact with T cells.


Analysis of the volatile organic compounds from leaves, flower spikes, and nectar of Australian grown Agastache rugosa.
The foraging choices of honey bees are influenced by many factors, such as floral aroma. The composition of volatile compounds influences the bioactivity of the aromatic plants and honey produced from them. In this study, Agastache rugosa was evaluated as part of a project to select the most promising medicinal plant species for production of bioactive honey. Headspace solid-phase microextraction HS-SPME /GC-MS was optimized to identify the volatile bioactive compounds in the leaves, flower spikes, and for the first time, the flower nectar of Australian grown A. rugosa. Methyl chavicol (= estragole) was the predominant headspace volatile compound in the flowers with nectar, flower spikes, and leaves, with a total of 97.16%, 96.74% and 94.35%, respectively. Current results indicate that HS-SPME/GC-MS could be a useful tool for screening estragole concentration in herbal products. Recently, estragole was suspected to be carcinogenic and genotoxic, according to the European Union Committee on Herbal Medicinal Products. Further studies are needed on safe daily intake of Agastache as herbal tea or honey, as well as for topical uses.


General competencies are intrinsic to emergency medicine training: a multicenter study.
The Accreditation Council for Graduate Medical Education (ACGME) has promulgated six areas called General Competencies (GCs) that residency programs are required to evaluate. The authors sought to determine if these domains were an intrinsic part of emergency medicine (EM) residency training by using a global assessment evaluation device. This was an observational, multicenter, cross-sectional study that compared GC acquisition between first-, second-, and third-year (EM1, EM2, and EM3) residents. Five postgraduate year (PGY) 1 to PGY 3 allopathic EM programs in Michigan participated. A global assessment form using a 1 through 9 ordinal scale with 86 scoring items was given to program directors for each resident in their programs. Analysis of variance (ANOVA) was used to compare the means between EM1, EM2, and EM3 scores. Five EM programs evaluated 150 residents. The GC scores were as follows: Patient Care: EM1 4.92, EM2 5.79, and EM3 6.40; Medical Knowledge: EM1 4.90, EM2 5.80, and EM3 6.46; Practice-based Learning and Improvement: EM1 4.60, EM2 5.48, and EM3 6.16; Interpersonal and Communication Skills: EM1 4.99, EM2 5.39, and EM3 6.01; Professionalism: EM1 5.43, EM2 5.68, and EM3 6.27; Systems-based Practice: EM1 4.80, EM2 5.48, and EM3 6.21. ANOVA showed statistically significant differences (p < 0.001) for all GCs. EM residents from several residency programs showed statistically significant progressive acquisition of the ACGME GCs using a global assessment device. This suggests that the GCs may be an intrinsic component in the training of EM residents.


Risk assessment as a practice method in child protective services.
This article offers an overview of the development of risk assessment and its purpose and goals, a summary of research on risk assessment to date, and suggestions regarding future research. This overview indicates that the use of risk assessment is a widespread phenomenon, with differing goals and purposes. Research on risk assessment confirms the lack of conceptual clarity and the difficulty of incorporating it into a child protective services system that emphasizes findings of abuse and/or neglect as the key to continuing involvement in the system.


Spectrum of imaging findings in hyperextension injuries of the neck.
Nonphysiologic hyperextension and lateral forces acting on the cervical spine and soft-tissue structures of the neck can result in a wide spectrum of injury patterns. Multiple factors (eg, patient age; the underlying morphologic features of the cervical spine; the magnitude, vector, and maximal focus of the force) all influence the observed patterns and the severity of injury. A review of the 5-year trauma database in two trauma centers revealed various injury patterns that were frequently recognized in patients with clinical evidence or historical documentation of a predominant hyperextension mechanism. Injuries included anterior arch avulsion and posterior arch compression fractures of the atlas, odontoid fractures, traumatic spondylolisthesis and teardrop fracture of C2, laminar and articular pillar fractures, and hyperextension dislocation injuries. More severe injuries were observed in patients with underlying predisposing conditions (eg, degenerative spondylosis, ankylosing spondylitis, diffuse idiopathic skeletal hyperostosis). Knowledge of the involved biomechanical factors provides a framework for understanding these injury patterns. Diagnostic imaging, especially computed tomography and magnetic resonance imaging, plays a fundamental role in the assessment of patients with suspected cervical injury. Furthermore, cross-sectional imaging facilitates the recognition of accompanying injuries to the face, the head, and the vascular structures of the neck.


New diterpene glycosides of the fungus Acremonium striatisporum isolated from a sea cucumber.
Three new diterpene glycosides, virescenosides O (1), P (2), and Q (3), have been isolated from a marine strain of Acremonium striatisporum KMM 4401 associated with the holothurian Eupentacta fraudatrix. Their structures were determined on the basis of HRMALDIMS and NMR data as beta-D-altropyranosido-19-isopimara-8(14),15-diene-7alpha,3beta-diol (1), beta-D-altropyranosido-19-7-oxoisopimara-8(9),15-diene-3beta-ol (2), and beta-D-mannopyranosido-19-isopimara-7,15-diene-3beta-ol (3). The cytotoxic activity of the virescenosides was examined.


[Closure of Botallo's ductus arteriosus with indomethacin. Considerations on 4 cases].
Persistence of Patent Ductus Arteriosus (PDA) frequently occurs in preterm infants. In a remarkable number of cases, PDA may undergo spontaneous closure. In other cases, especially in patients with respiratory problems, a ductal left-to-right shunt may compromise the cardiac-respiratory function. Until a few years ago, surgery was the only possible therapy. Recently, pharmacological closure of the duct by means of Indomethacin, a powerful inhibitor of prostaglandin synthesis, was successfully carried out. The medical treatment, however, appears to be less effective if it is carried out on patients older than 2 weeks. It has also been reported that in patient with a very low birthweight and with lack of muscular tissue in the duct wall, closure of the duct may be transient. It is the purpose of this communication to report observations made on 4 preterm infants of 24-33 weeks gestational age, and with a weight range of 660 to 2.280 g. They received Indomethacin via naso-gastric tube, in a single dose of 0,3 mg/Kg, within 8 days of age. Successful closure of the PDA was obtained in all cases. The pharmacological treatment resulted also in reductions in ventilatory support requirements, reduced respiratory component of acidosis, reduced pCO2, increased pO2. A second administrations of Indo was necessary only in one case, and it was followed by permanent closure of the duct. Collateral effects, such as reduced diuresis and reduced number of platelets, were transient. Indo therapy was successful regardless of very low birthweight: in three cases, birthweight was under 1.000 g.(ABSTRACT TRUNCATED AT 250 WORDS)


De Novo Biosynthesis of Free Vaccenic Acid with a Low Content of Oleic Acid in Saccharomyces cerevisiae.
Omega-7 (ω-7) fatty acids have potential application in the fields of nutraceutical, agricultural, and food industry. The natural ω-7 fatty acids are currently from plants or vegetable oils, which are unsustainable and limited by the availability of plant sources. Here, we developed an innovative biosynthetic route to produce vaccenic acid (C18:1 ω-7) while minimizing oleic acid (C18:1 ω-9) content in Saccharomyces cerevisiae. We have engineered S. cerevisiaeto produce C18:1 ω-7 by expressing a fatty acid elongase from Rattus norvegicus. To reduce the content of C18:1 ω-9, the endogenous desaturase Ole1 was replaced by the desaturase, which has specific activity on palmitoyl-coenzyme A (C16:0-CoA). Finally, the production of free C18:1 ω-7 was improved by optimizing the source of cytochrome b5 and overexpressing endoplasmic reticulum chaperones. After combining these strategies, the yield of C18:1 ω-7 was increased from 0 to 9.3 mg/g DCW and C18:1 ω-9 was decreased from 25.2 mg/g DCW to 1.6 mg/g DCW. This work shows a de novo synthetic pathway to produce the highest amount of free C18:1 ω-7 with a low content of C18:1 ω-9 in S. cerevisiae.


Minimizing the Risk of Catastrophic Health Expenditure in China: A Multi-Dimensional Analysis of Vulnerable Groups.
Background: In moving toward universal health coverage in China, it is crucial to identify which populations should be prioritized for which interventions rather than blindly increasing welfare packages or capital investments. We identify the characteristics of vulnerable groups from multiple perspectives through estimating catastrophic health expenditure (CHE) and recommend intervention priorities. Methods: Data were from National Health Service Survey conducted in 2003, 2008, and 2013. According to the recommendation of WHO, this study adopted 40% as the CHE threshold. A binary regression was used to identify the determinants of CHE occurrence; a probit model was used to obtain CHE standardized incidence under the characteristics of single and two dimensions in 2013. Results: The total incidence of CHE in 2013 was 13.9%, which shows a general trend of growth from 2003 to 2013. Families in western and central regions and rural areas were more at risk. Factors related to social demography show that households with a female or an unmarried head of household or with a low socioeconomic status were more likely to experience CHE. Households with older adults aged 60 and above had 1,524 times higher likelihood of experiencing CHE. Among the health insurance schemes, the participants covered by the New Rural Cooperative Medical Scheme had the highest risk compared with the participants of all basic health insurance schemes. Households with several members seeking outpatient, inpatient care or with non-communicable diseases were more likely to experience CHE. Households with members not seeing a doctor or hospitalized despite the need for it were more likely to experience CHE. Characteristics such as a household head with characteristics related to low socioeconomic status, having more than two hospitalized family members, ranked high. Meanwhile, the combination of having illiterate household heads and with being covered by other health insurance plans or by none ranked the first place. Cancer notably caused a relatively high medical expenditure among households with CHE. Conclusion: In China, considering the vulnerability of the population across different dimensions is conducive to the alleviation of high CHE. Furthermore, people with multiple vulnerabilities should be prioritized for intervention. Identifying and targeting them to offer help and support will be an effective approach.


Assessing the effect of chronic oedema with associated ulceration.
Chronic oedema is a common problem in the UK and, given the country's ageing population, the numbers are predicted to rise. In an epidemiological study carried out in Derby, England, researchers found the prevalence of chronic oedema to be 3.99 in every 1000 people, with the prevalence increasing to 10.31 in those aged 65-74 years. Often, patients with untreated chronic oedema will develop an ulceration that can lead to further costs and hospital admissions. The cost of treating chronic wounds has been estimated at £2.3 billion-£3.1 billion a year. It is therefore surprising that given the number of patients living with these problems, there is still a lack of knowledge and skill among nurses when assessing patients with chronic oedema and associated ulceration. This article offers advice for nurses when assessing leg ulcers in patients with chronic oedema, detailing the visual skin changes most frequently seen in these patients. The article also discusses some of the treatment options available, briefly covering the advantages and disadvantages of each option.


Design and Synthesis of a Fluorescent Probe with a Large Stokes Shift for Detecting Thiophenols and Its Application in Water Samples and Living Cells.
A turn-on florescent probe (probe-KCP) was developed for highly selective detection of thiophenols based on a donor-excited photo-induced electron transfer mechanism. Herein, the synthesis of the probe, a chalcone derivative, through a simple straightforward combination of a carbazole-chalcone fluorophore with a 2,4-dinitrophenyl functional group. In a kinetic study of the probe-KCP for thiophenols, the probe displayed a short response time (~30 min) and significant fluorescence enhancement. In selection and competition experiments, the probe-KCP exhibited excellent selectivity for thiophenols over glutathione (GSH), cysteine (Cys), sodium hydrosulfide (NaSH), and ethanethiol (C₂H₅SH) in addition to common anions and metal ions. Using the designed probe, we successfully monitored and quantified thiophenols, which are highly toxic. This turn-on fluorescence probe features a remarkably large Stokes shift (130 nm) and a short response time (30 min), and it is highly selective and sensitive (~160-fold) in the detection of thiophenols, with marked fluorescence in the presence of thiophenols. probe-KCP responds to thiophenols with a good range of linearity (0⁻15 μM) and a detection limit of 28 nM (R² = 0.9946) over other tested species mentioned including aliphatic thiols, thiophenol analogues, common anions, and metal ions. The potential applications of this carbazole-chalcone fluorescent probe was successfully used to determine of thiophenols in real water samples and living cells with good performance and low cytotoxicity. Therefore, this probe has great potential application in environment and biological samples.


Enzyme-linked immunosorbent assay to detect Lawsonia intracellularis in rabbits with proliferative enteropathy.
Lawsonia intracellularis is an obligate intracellular pathogenic bacterium that causes proliferative enteropathy in domestic and experimental animals. Antiserum against synthetic peptides of the Lawsonia surface antigen (LsaA) well recognized L. intracellularis in infected ileum by immunohistochemistry. The synthetic peptides in LsaA showed strong reaction with serum from rabbits infected with L. intracellularis by enzyme-linked immunosorbent assay. These results suggest that ELISA used synthetic peptides in LsaA and anti-LsaA serum might be useful to diagnose for proliferative enteropathy.


Autologous Platelet Collection and Storage to Support Thrombocytopenia in a Leukemia Patient with Platelet Alloimmunization Undergoing Chemotherapy.
Platelet alloimmunization occurs frequently in multitransfused patients. To prevent the posttransfusion complications donor white blood cells (WBCs) must be removed from the platelet concentrates (PC). But sometimes the centrifugation and filtration of WBC is not enough and in such conditions HLA-matched PC are recommended. Cryopreservation of autologous PC offers a potential solution to this problem. We investigated the feasibility of supporting the aplastic period after chemotherapy in a 54-year-old leukemia patient with autologous platelets collected by apheresis and cryopreserved. Our case report demonstrates that autologous apheresis platelet transfusion might be a feasible approach to high risk patients with bleeding tendencies.


Annual review of section on ocular disease: diseases of the optic nerve, tracts, and visual cortex.
The ophthalmic literature dealing with diseases of the optic nerve, tracts, and visual cortex was reviewed for the period November 1977 to December 1978. Fifty-nine papers of interest to optometrists have been abstracted. Main areas of interest include: role of axoplasmic transport in production of disc edema and cotton-wool spots; ocular effects of local and systemic vascular disturbances; congenital abnormalities of the eye alone and in association with other bodily malformation; field testing, including an ingenious ophthalmoscopic method which can lead to detection of very small scotomas; a new quantitative method for assessing integrity of the optic nerve in multiple sclerosis and related conditions; stimulating questions on the development of optic atrophy and on its relation to retinal vascularity; considerations of acquired and congenital abnormalities of the cavernous sinus and their ocular manifestations; swinging-flashlight test in hemianoptic patients; a case of subacute sclerosing panecephalitis; methods to enhance magnification during opthalmoscopy; methods enabling a closer investigation of the retinal nerve fiber layer.


Determinants of termination of breastfeeding within the first 2 years of life in India: evidence from the National Family Health Survey-2.
The present study assesses socio-demographic and health service determinants of termination of breastfeeding within the first 2 years of life in India by analysing data from the nationally representative National Family Health Survey-2 using Cox regression modelling techniques. While the likelihood of stopping breastfeeding increased with increasing household wealth status, it declined with increasing maternal age at childbirth. The likelihood of stopping breastfeeding was significantly higher among female children compared with male children, and the gender differential was attenuated by increasing maternal educational status. Overall, findings of the present study suggest that breastfeeding promotion programmes in India should focus on certain high-risk mother-child pairs such as female infants, first-born babies, babies born in the private sector and in urban areas, as well as mothers who are literate, have a higher wealth status, are aged less than 20 years and belong to Sikh or Christian communities. Qualitative studies to understand cultural factors or norms and causal pathways responsible for the association of identified factors and early termination of breastfeeding, especially household wealth status and maternal education, are also called for.


Evidence for an extended duration of GABA-mediated excitation in the developing male versus female hippocampus.
Gamma-aminobutyric acid (GABA) is as an excitatory neurotransmitter during brain development. Activation of GABA(A) receptors in neonatal rat hippocampus results in chloride efflux and membrane depolarization sufficient to open voltage sensitive calcium channels. As development progresses, there is a decline in the magnitude of calcium influx subsequent to GABA(A) receptor activation and the number of cells that respond to GABA with excitation. By the second postnatal week in the rat, GABA action in the hippocampus is predominantly inhibitory. The functional consequences and endogenous regulation of developmental GABA-mediated excitation remains under-explored. Hippocampal neurons in the newborn male and female rat respond to GABA(A) receptor activation with increased intracellular calcium and are susceptible to GABA-mediated damage -- both being indicative of the excitatory nature of GABA. In the present study we observed that by postnatal day 7, only males are susceptible to GABA(A) agonist-induced damage and respond to GABA(A) agonist administration with elevated levels of intracellular calcium in cultured hippocampal neurons. By postnatal day 14, GABA(A) agonist administration was without effect on intracellular calcium in both males and females. The age-related sex difference in the impact of GABA(A) receptor activation correlates with a sex difference in chloride co-transporter expression. Males have elevated protein levels of pNKCC1 on PN0 and PN7, with no sex difference by PN14. In contrast, females displayed elevated levels of KCC2 on PN7. This converging evidence infers that sex affects the duration of GABA(A) receptor-mediated excitation during normal hippocampal development, and provides a mechanism by which the effect is mediated.


Activity-dependent neurotransmitter-receptor matching at the neuromuscular junction.
Signaling in the nervous system requires matching of neurotransmitter receptors with cognate neurotransmitters at synapses. The vertebrate neuromuscular junction is the best studied cholinergic synapse, but the mechanisms by which acetylcholine is matched with acetylcholine receptors are not fully understood. Because alterations in neuronal calcium spike activity alter transmitter specification in embryonic spinal neurons, we hypothesized that receptor expression in postsynaptic cells follows changes in transmitter expression to achieve this specific match. We find that embryonic vertebrate striated muscle cells normally express receptors for glutamate, GABA, and glycine as well as for acetylcholine. As maturation progresses, acetylcholine receptor expression prevails. Receptor selection is altered when early neuronal calcium-dependent activity is perturbed, and remaining receptor populations parallel changes in transmitter phenotype. In these cases, glutamatergic, GABAergic, and glycinergic synaptic currents are recorded from muscle cells, demonstrating that activity regulates matching of transmitters and their receptors in the assembly of functional synapses.


Hypomethylation of MB-COMT promoter is a major risk factor for schizophrenia and bipolar disorder.
The variability in phenotypic presentations and the lack of consistency of genetic associations in mental illnesses remain a major challenge in molecular psychiatry. Recently, it has become increasingly clear that altered promoter DNA methylation could play a critical role in mediating differential regulation of genes and in facilitating short-term adaptation in response to the environment. Here, we report the investigation of the differential activity of membrane-bound catechol-O-methyltransferase (MB-COMT) due to altered promoter methylation and the nature of the contribution of COMT Val158Met polymorphism as risk factors for schizophrenia and bipolar disorder by analyzing 115 post-mortem brain samples from the frontal lobe. These studies are the first to reveal that the MB-COMT promoter DNA is frequently hypomethylated in schizophrenia and bipolar disorder patients, compared with the controls (methylation rate: 26 and 29 versus 60%; P=0.004 and 0.008, respectively), particularly in the left frontal lobes (methylation rate: 29 and 30 versus 81%; P=0.003 and 0.002, respectively). Quantitative gene-expression analyses showed a corresponding increase in transcript levels of MB-COMT in schizophrenia and bipolar disorder patients compared with the controls (P=0.02) with an accompanying inverse correlation between MB-COMT and DRD1 expression. Furthermore, there was a tendency for the enrichment of the Val allele of the COMT Val158Met polymorphism with MB-COMT hypomethylation in the patients. These findings suggest that MB-COMT over-expression due to promoter hypomethylation and/or hyperactive allele of COMT may increase dopamine degradation in the frontal lobe providing a molecular basis for the shared symptoms of schizophrenia and bipolar disorder.


Pharmacovigilance and principle of nonmaleficence in sex reassignment.
Physicians are obliged to provide treatment that is consistent with their commitment to avoid or minimize harm (nonmaleficence) and their commitment to do good (beneficence). Therefore, if patient's desires were contradictory to the primary aim of medicine, the doctor's calling would require him/her to thoroughly analyze the cause of the disease and provide an adequate as well as ethical treatment rather than obediently follow patient's requests. Yet, chemical and surgical sex reassignment is one of the areas where some physicians surrender to the desire of their patients instead of finding out what their real condition is and trying to manage it in a way the essence of medicine would require. The objective of this article was to provide specific pharmacovigilance search details for the evaluation of the current situation and the scientific background of the treatment of gender dysphoria and to analyze its conformity with one of the two main ethical principles of medicine - nonmaleficence. Literature retrieval was accessed through Medline (1979-2011) using the terms "gender dysphoria," "replacement hormonal therapy," and "pharmacovigilance." The article concludes that hormonal and surgical interventions have not proven to be medically justified and could be harmful, not treating the cause, but resulting in irreversible disability. Thus, these interventions contradict the principle of nonmaleficence and goals of basic therapeutics and pharmacovigilance. They are not based on clinical trials and are lacking a thorough follow-up assessment.


The Relationship Between MMP-2 -1306C>T and MMP-9 -1562C>T Polymorphisms and the Risk and Prognosis of T-Cell Acute Lymphoblastic Leukemia in a Chinese Population: A Case-Control Study.
T-cell acute lymphoblastic leukemia (T-ALL) is a malignant hematological disease and is often accompanied by a variety of genetic abnormalities. Hence, our study aims to investigate the relationship between MMP-2 -1306C>T and MMP-9 -1562C>T polymorphisms and the risk and prognosis of T-ALL. From April 2009 to February 2011, a total of 376 T-ALL patients were chosen as the case group. Meanwhile, 352 healthy people who passed routine health examinations were selected as the control group. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was used to detect the frequency of MMP-2 -1306C>T (rs243865) and MMP-9 -1562C>T (rs3918242) polymorphisms in the study subjects. The serum levels of MMP-2 and MMP-9 were detected using enzyme-linked immunosorbent assay (ELISA). A Kaplan-Meier analysis was employed to analyze the event-free survival (EFS) rates of the T-All patients with different MMP-2 and MMP-9 genotypes. A multivariate COX model was applied to analyze the relationship between MMP-2 and MMP-9 polymorphisms and the prognosis of T-ALL patients. A C-statistic and net reclassification index (NRI) was carried out to evaluate the predictive value of MMP-2 and MMP-9 gene polymorphisms using the Cox model. Compared to the control group, the genotypic frequency of MMP-2 -1306C>T (CT + TT) and MMP-9 -1562C>T (CT + TT) in the case group was significantly higher. The serum level of MMP-9 was markedly elevated in T-ALL patients with the CT + TT genotype compared to patients with the CC genotype. The results of the Kaplan-Meier analysis showed that the median EFS was lower in T-ALL patients with the CT + TT genotype of MMP-9 -1562C>T compared to patients with the CC genotype. The results of a multivariate analysis using the Cox proportional hazard model indicated that the MMP-9 -1562C>T polymorphism was associated with the prognosis of T-ALL patients. These results indicated that MMP-2 -1306C/T and MMP-9 -1562C/T polymorphisms might be associated with an increased risk of T-ALL. The MMP-9 -1562C>T polymorphism may also be related to the prognosis of T-ALL patients.


Lymphocyte/Monocyte Ratio is a Novel Predictor for Early Stage Extranodal Natural Killer/T-cell Lymphoma, Nasal Type.
Objective: Great heterogeneity exists in clinical behavior and survival outcome in patients with stage IE/IIE extranodal natural killer/T-cell lymphoma, nasal type (ENKTL). In this study, we proposed lymphocyte/monocyte ratio (LMR) as a new prognostic factor for these early stage ENKTL. Methods: We retrospectively examined the LMR as a prognostic variable in a cohort of 379 patients with newly diagnosed stage IE/IIE ENKTL. The relationship between the LMR and clinicopathologic variables were analyzed in Kaplan-Meier log-rank survival analysis, and the Cox proportional hazards model was used to determine the survival significance of the LMR for both progression-free survival (PFS) and overall survival (OS). Results: Patients were categorized into two different groups based on the LMR using cut-off value of 2.0. The 5-year PFS rates in the low and high LMR group were 43.9% and 62.7%, respectively, and the 5-year OS rates in the two groups were 59.1% and 77.7%, respectively. In multivariate analysis, low LMR at diagnosis was associated with worse PFS (hazard ratio 1.611, 95% confidence interval: 1.027-2.525, P =0.038) independent of age (P=0.033) and treatment stratagem (P<0.001), and indicated worse OS (hazard ratio 2.003, 95% confidence interval: 1.124-3.569, P =0.018) independent of age (P=0.007), LDH level (P=0.042), local tumor invasiveness (P=0.008), and treatment stratagem (P<0.001). Conclusion: The LMR is an independent prognostic factor for both DFS and OS in patients with stage IE/IIE ENKTL, and provides additional prognostic value beyond standard clinicopathological parameters.


Entropy contribution toward micelle-driven deintercalation of drug-DNA complex.
Micelle-assisted "deintercalation" of intercalated drug/mutagen molecules from DNA is a well-established phenomenon; however, the driving energy cost for such a process is still not properly understood. In the present contribution, we have estimated the various energetic parameters for the SDS micelle-assisted deintercalation of a model DNA intercalator phenosafranine (PSF) using isothermal titration calorimetry (ITC) measurement. Both steady-state and picosecond-resolved fluorescence measurements provide strong evidence for the relocation of PSF molecules from the DNA interior to the micellar interface at an SDS concentration above cmc. The overall deintercalation process has been found to be enthalpy-wise forbidden (endothermic); however, it is strongly favored by a high positive entropy change, which can be correlated with the change in the associated hydration structure at the macromolecular interface.


[Magnetic Resonance Imaging in Facial Bell's Palsy].
The meaning of facial nerve enhancement by gadolinium-enhanced magnetic resonanca imaging (MRI) in Bell's palsy is very discussed. Aim of our study was to define the radiological data in facial Bell's palsy and to correlate them with clinical data. 34 patients with facial Bell's palsy were studied prospectively. Each patient had two gadolinium-enhanced MRI. The first MRI was realized during the first ten days. The second MRI was realized three months later. Two patients with facial nerve enhancement localized on the third portion was not clinically normalized at 3 months. Facial nerve enhancement and his precise location was noted. Facial nerve enhancement was noted in 71,5% on first MRI and in 30% on second MRI. Gadolinium-enhancement was predominantly noted on the Fundus. No patients with facial nerve enhancement persisting at three months was clinically normalized at one month. Facial nerve enhancement localized on the fundus and/or on the third portion, persisting at three months, could be consider as a negative landmark of fast clinical facial recuperation.


Couples' approaches to changes in everyday life during the first year after stroke.
The aim of the study was to identify and describe two couples' approaches to changes in everyday life during the first year after a stroke. An additional aim was to describe how the couples viewed rehabilitation as well as their own personal training relative to changes in everyday life during the first year at home after stroke. The study design was a prospective longitudinal case study based on two couples where one of the spouses in each couple had experienced a stroke. Data collection consisted of interviews and a questionnaire and took place in the participants' homes. Data analysis utilized a constant comparative method. The findings showed a divergence in the couples' approaches to changes in their everyday life at home and were described through the following categorizations: engaging in occupations, getting experience and thereby feedback from doing, changing one's occupational needs and demands, contributing to a picture of a possible future and, integrating training in everyday life. Getting experience and feedback from doing was found to be a key category or "driver" in the change process. The couples' experiences of changes in everyday life after stroke illustrated two very divergent approaches, which is discussed in the paper. The approaches in turn had consequences for how daily life was spent after stroke which is also discussed.


Adipokinetic hormone activities in insect body infected by entomopathogenic nematode.
The role of adipokinetic hormone (AKH) in the firebug Pyrrhocoris apterus adults infected by the entomopathogenic nematode (EPN) Steinernema carpocapsae was examined in this study. It was found that co-application of EPN and AKH enhanced firebug mortality about 2.5 times within 24h (from 20 to 51% in EPN vs. EPN+AKH treatments), and resulted in metabolism intensification, as carbon dioxide production in firebugs increased about 2.1 and 1.6times compared to control- and EPN-treated insects, respectively. Accordingly, firebugs with reduced expression of AKH receptors showed a significantly lower mortality (by 1.6 to 2.9-folds), and lower general metabolism after EPN+AKH treatments. In addition, EPN application increased Akh gene expression in the corpora cardiaca (1.6times), AKH level in the corpora cardiaca (1.3times) and haemolymph (1.7times), and lipid and carbohydrate amounts in the haemolymph. Thus, the outcomes of the present study demonstrate involvement of AKH into the anti-stress reaction elicited by the nematobacterial infection. The exact mechanism by which AKH acts is unknown, but results suggested that the increase of metabolism and nutrient amounts in haemolymph might play a role.


The Association of rs4753426 Polymorphism in the Melatonin Receptor 1B (MTNR1B) Gene and Susceptibility to Adolescent Idiopathic Scoliosis: A Systematic Review and Meta-analysis.
Adolescent idiopathic scoliosis (AIS) is a tridimensional structural deformity of the spine that may deteriorate progressively, leading to significant functional limitations and pain problems. Several previous studies have implicated the rs4753426 single nucleotide polymorphism in the melatonin receptor 1B (MTNR1B) gene in the etiology of AIS. However the sample sizes were limited and the findings of those studies were inconsistent. An overall assessment of the evidence supporting this association has not been previously conducted. To provide a comprehensive assessment and synthesis of the currently available evidence on the association between rs4753426 and AIS. A systematic review and meta-analysis. University hospital, China. This review followed the Preferred Reporting Items for Systematic Review and Meta-Analyses guidelines. PubMed (MEDLINE), EMBASE, Scopus databases, and WANFANG databases were systematically searched through December 2014 to identify relevant studies following a sensitive strategy. Statistical analysis was performed using the Review Manager 5.2 software. Summary odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were estimated using the fixed-effect inverse variance model for allelic (C vs. T) and genotypic comparisons. Four papers including 5 studies which involved 2,552 AIS cases and 2,738 controls were identified for this meta-analysis. The results showed that C allele of the rs4753426 was significantly associated with AIS (OR = 1.12, 95% CI: 1.03-1.21, P = 0.01). CT and CC genotypes were 26% (OR = 1.26, 95% CI: 1.04-1.53, P = 0.01) and 28% (OR = 1.28, 95% CI: 1.05-1.56, P = 0.01), respectively, more likely to have AIS compared with CC genotype. As for the dominant model (CC+TT vs. TT), summary ORs showed statistically significant association with AIS (OR = 1.28, 95% CI: 1.06-1.53, P = 0.009). Compared with the CT+TT genotype, the summary ORs of the CC genotype showed marginally statistically significant association with AIS (OR = 1.11, 95 % CI: 0.99-1.24, P = 0.07). The subgroup meta-analysis results showed the C allele and each genotype were significantly associated with AIS in the Asian group but not in the Caucasian group. Paucity of available literature. To our knowledge, there has been no meta-analysis to analyze the association between rs4753426 polymorphism in the MTNR1B gene and AIS. This systematic review was a comprehensive analysis of the currently available evidence, and found an overall significant association of rs4753426 polymorphism with the risk of AIS, especially in the Asian population. Further investigation of this association is necessary in other populations.


Conjugate addition of amines to an alpha,beta-unsaturated imine derived from alpha-aminophosphonate. Synthesis of gamma-amino-alpha-dehydroaminophosphonates.
Aza-Michael reaction of ammonia, aliphatic, aromatic and optically active amines to an alpha,beta-unsaturated imine derived from alpha-aminophosphonate affords alpha-dehydroaminophosphonates with a gamma-stereogenic center bearing an amino group. Resulting gamma-amino alpha-dehydroaminophosphonates can be used for the preparation of phosphorylated pyrimidine derivatives.


Physico-chemical characterization of growth hormone from water buffaloes (Bubalus bubalis).
A purified preparation of growth hormone from pituitaries of water buffaloes (Bubalus bubalis) has been extensively characterized with regard to physico-chemical properties. The molecular size of buffalo GH (buGH) by electrospray ionization mass spectroscopy (ES-MS) was found to be 21394.00+/-8.44Da and its stokes radius was determined as 2.3 nm. Size heterogeneity in buffalo GH was checked both by electrophoresis and molecular sieve chromatography using 125I-labelled buffalo GH. Similar size heterogeneity was found in standard preparations of ovine and bovine growth hormones. Isoelectric focussing and chromatofocussing indicated charge heterogeneity in buffalo GH preparation. Major charge isoforms having pI of 7.2, 7.7 and minor forms in the pI range of 5.7 to 7.0 were found. Lectin chromatography on Concanavalin A matrix showed that less than 1% of buffalo GH was glycosylated. Heterogeneity in NH2-terminal sequence was also observed, with alanine, phenylalanine and methionine as the NH2-terminal residues as checked by dansyl and DABITC methods. Estimation of tryptophan residue indicated that a single tryptophan residue was present. Ellman's method showed presence of two disulfide bridges per mole of buffalo GH. Intrinsic fluorescence spectrum of buffalo GH exhibited lambda emission maximum at 337 nm. UV-CD spectrum showed that almost 48% of the secondary structure of buGH was constituted by alpha-helicity. The T(M) of buGH as determined by differential scanning calorimetric (DSC) studies was found to be 63 degrees C.


Bacterial motility on a surface: many ways to a common goal.
When free-living bacteria colonize biotic or abiotic surfaces, the resultant changes in physiology and morphology have important consequences on their growth, development, and survival. Surface motility, biofilm formation, fruiting body development, and host invasion are some of the manifestations of functional responses to surface colonization. Bacteria may sense the growth surface either directly through physical contact or indirectly by sensing the proximity of fellow bacteria. Extracellular signals that elicit new gene expression include autoinducers, amino acids, peptides, proteins, and carbohydrates. This review focuses mainly on surface motility and makes comparisons to features shared by other surface phenomenon.


The epidemiology of primary headache disorders.
Primary headache disorders encompass a heterogeneous group of neurologic disorders that cause recurrent or persistent head pain without any clear underlying cause. The epidemiology of the most common primary headache disorders, tension-type headache and migraine, the most common trigeminal autonomic cephalalgia, cluster headache, as well as chronic daily headache are addressed in this review. The incidence, prevalence, burden, and comorbidities of these disorders are discussed, and the risk factors for progression from episodic to chronic daily headache are also reviewed.


A functional in vitro cell-free system for studying DNA repair in isolated nuclei.
Assessment of DNA repair is an important endpoint measurement when studying the biochemical mechanisms of the DNA damage response and when investigating the efficacy of chemotherapy, which often uses DNA-damaging compounds. Numerous in vitro methods to biochemically characterize DNA repair mechanisms have been developed so far. However, such methods have some limitations, which are mainly due to the lack of chromatin organization in the DNA templates used. Here we describe a functional cell-free system to study DNA repair synthesis in vitro, using G1-phase nuclei isolated from human cells treated with different genotoxic agents. Upon incubation in the corresponding damage-activated cytosolic extracts, containing biotinylated dUTP, nuclei were able to initiate DNA repair synthesis. The use of specific DNA synthesis inhibitors markedly decreased biotinylated dUTP incorporation, indicating the specificity of the repair response. Exogenously added human recombinant PCNA protein, but not the sensors of UV-DNA damage DDB2 and DDB1, stimulated UVC-induced dUTP incorporation. In contrast, a DDB2PCNA- mutant protein, unable to associate with PCNA, interfered with DNA repair synthesis. Given its responsiveness to different types of DNA lesions, this system offers an additional tool to study DNA repair mechanisms.This article has an associated First Person interview with the first author of the paper.


Development of a novel magnetic particle-based agglutination immunoassay for anticardiolipin antibody detection in syphilis.
Serological tests of non-treponemal and treponemal types are the most frequently used for syphilis diagnosis. Treponemal tests are available in wide variety of assay formats; however, limited advances have been made for the improvement of conventional non-treponemal tests. The objective of this work was to develop a novel non-treponemal magnetic particle-based agglutination assay (NT-MAA) and evaluate its feasibility for syphilis testing. Cardiolipin was modified and coupled to magnetic microbeads. Serum diluted in phosphate-buffered saline was mixed with cardiolipin-coupled beads and incubated in a round bottom microplate for 90-120 min followed by visual inspection. A panel of reported syphilis (n=127) and non-reactive (n=244) specimens was prepared to evaluate the NT-MAA performance in comparison to conventional rapid plasma reagin (RPR). Treponema pallidum particle agglutination (TP-PA) assay and enzyme immunoassay (EIA) were included. Analytical sensitivity and reproducibility of NT-MAA were also determined. The non-treponemal NT-MAA and RPR showed sensitivity of 90.6% and 88.2% and specificity of 96.7% and 100%, respectively. The treponemal TP-PA and EIA yielded sensitivity of 100% and 99.2%, respectively, and 100% specificity by both assays. The per cent agreement between NT-MAA and RPR was 97% (kappa=0.931, 95% CI 0.891 to 0.971). Analytical sensitivity determined with IgM anticardiolipin antibody (ACA) was 2.6 µg/mL for both NT-MAA and RPR, while IgG ACA yielded 0.9 µg/mL and 1.7 µg/mL for NT-MAA and RPR, respectively. Qualitative results of intra-assay and interassay reproducibility revealed 100% consistency for NT-MAA. Preliminary evaluation of the novel NT-MAA validated proof of concept using laboratory-characterised syphilis sera and demonstrated performance comparable to RPR. Further validation of NT-MAA using additional specimens with better clinical staging may broaden the scope of developed test for syphilis diagnosis.


Mephedrone (4-Methylmethcathinone): Acute Behavioral Effects, Hyperthermic, and Pharmacokinetic Profile in Rats.
Mephedrone (MEPH) is a synthetic cathinone derivative with effects that mimic MDMA and/or cocaine. Our study in male Wistar rats provides detailed investigations of MEPH's and its primary metabolite nor-mephedrone's (nor-MEPH) pharmacokinetics and bio-distribution to four different substrates (serum, brain, lungs, and liver), as well as comparative analysis of their effects on locomotion [open field test (OFT)] and sensorimotor gating [prepulse inhibition of acoustic startle reaction (PPI ASR)]. Furthermore, in order to mimic the crowded condition where MEPH is typically taken (e.g., clubs), the acute effect of MEPH on thermoregulation in singly- and group-housed rats was evaluated. Pharmacokinetics of MEPH and nor-MEPH after MEPH (5 mg/kg, sc.) were analyzed over 8 h using liquid chromatography with mass spectrometry. MEPH (2.5, 5, or 20 mg/kg, sc.) and nor-MEPH (5 mg/kg, sc.) were administered 5 or 40 min before the behavioral testing in the OFT and PPI ASR; locomotion and its spatial distribution, ASR, habituation and PPI itself were quantified. The effect of MEPH on rectal temperature was measured after 5 and 20 mg/kg, sc. Both MEPH and nor-MEPH were detected in all substrates, with the highest levels detected in lungs. Mean brain: serum ratios were 1:1.19 (MEPH) and 1:1.91 (nor-MEPH), maximum concentrations were observed at 30 min; at 2 and 4 h after administration, nor-MEPH concentrations were higher compared to the parent drug. While neither of the drugs disrupted PPI, both increased locomotion and affected its spatial distribution. The effects of MEPH were dose dependent, rapid, and short-lasting, and the intensity of locomotor stimulant effects was comparable between MEPH and nor-MEPH. Despite the disappearance of behavioral effects within 40 min after administration, MEPH induced rectal temperature elevations that persisted for 3 h even in singly housed rats. To conclude, we observed a robust, short-lasting, and most likely synergistic stimulatory effect of both drugs which corresponded to brain pharmacokinetics. The dissociation between the duration of behavioral and hyperthermic effects is indicative of the possible contribution of nor-MEPH or other biologically active metabolites. This temporal dissociation may be related to the risk of prolonged somatic toxicity when stimulatory effects are no longer present.


Guidelines for humeral subluxation cutoff values: a comparative study between conventional, reoriented, and three-dimensional computed tomography scans of healthy shoulders.
The humeral subluxation index (HSI) is frequently assessed on computed tomography (CT) scans in conditions of the shoulder characterized by humeral displacement. An arbitrarily set HSI cutoff value of 45% for anterior subluxation and 55% for posterior subluxation has been widely accepted. We studied whether mean values and thresholds of humeral subluxation, in relation to the glenoid and scapula, were influenced by different imaging modalities. The HSIs referenced to the scapula (SHSI) and glenoid (GHSI) were compared between conventional CT scans, CT scans reoriented into the corresponding reference plane (ie, scapular plane for the SHSI and glenoid center plane for the GHSI), and 3-dimensional (3D) CT reconstructions of 120 healthy shoulders. The 95% normal range determined the cutoff values of humeral subluxation. The SHSI thresholds for conventional, reoriented, and 3D CT scans were 33%-61%, 44%-68%, and 49%-61%, respectively. A different mean SHSI was found for each imaging modality (conventional, 47%; reoriented, 56%; 3D, 55%; P ≤ .014), with the conventional SHSI showing an underestimation in 89% of the cases. GHSI thresholds for conventional, reoriented, and 3D CT scans were 40%-61%, 44%-56%, and 46%-54%, respectively. The mean GHSI did not differ between each imaging modality (conventional, 51%; reoriented, 50%; 3D, 50%; P = .146). The SHSI and GHSI are susceptible to different imaging modalities with consequently different cutoff values. The redefined HSI cutoff values guide physicians in the evaluation of humeral subluxation in conditions characterized by humeral displacement, depending on the available image data.


Actin filament barbed end elongation with nonmuscle MgATP-actin and MgADP-actin in the presence of profilin.
We have quantitated the in vitro interactions of profilin and the profilin-actin complex (PA) with the actin filament barbed end using profilin and nonmuscle beta,gamma-actin prepared from bovine spleen. Actin filament barbed end elongation was initiated from spectrin seeds in the presence of varying profilin concentrations and followed by light scattering. We find that profilin inhibits actin polymerization and that this effect is much more pronounced for beta,gamma-actin than for alpha-skeletal muscle actin. Profilin binds to beta,gamma-actin filament barbed ends with an equilibrium constant of 20 microM, decreases the filament elongation rate by blocking addition of actin monomers, and increases the dissociation rate of actin monomers from the filament end. PA containing bound MgADP supports elongation of the actin filament barbed end, indicating that ATP hydrolysis is not necessary for PA elongation of filaments. Initial analysis of the energetics for these reactions suggested an apparent greater negative free energy change for actin filament elongation from PA than elongation from monomeric actin. However, we calculate that the free energy changes for the two elongation pathways are equal if the profilin-induced weakening of nucleotide binding to actin is taken into consideration.


Stress reduction with heart transplant patients and their families: a multidisciplinary approach.
Stress reduction is an important part of the treatment plan for heart transplant patients at Loyola University Medical Center, Maywood, Illinois. This individualized stress reduction program is offered to both patient and family members through the complementary efforts of the social work and occupational therapy departments. The objectives of this program are to educate the participants in a variety of coping and stress reducing techniques to help prepare them for the multitude of stressors encountered during the entire transplant experience. Patients and family members have positively responded to this program, both verbally and in practice.


QM/MM excited state molecular dynamics and fluorescence spectroscopy of BODIPY.
Absorption and emission spectra arising from the lowest energy transition in BODIPY have been simulated in the gas phase and water using a quantum mechanics/molecular mechanics (QM/MM) approach. Kohn-Sham density functional theory (DFT) is used to calculate both ground (So) and first excited (S1) states using the maximum overlap method to obtain the S1 state. This approach gives ground and excited state structures in good agreement with structures found using multiconfigurational perturbation theory (CASPT2). Application of a post-self-consistent field spin-purification relationship also yields transition energies in agreement with CASPT2 and available experimental data. Spectral bands were simulated using many structures taken from ab initio molecular dynamics simulations of the ground and first excited states. In these simulations, DFT is used for BODIPY, and in the condensed phase simulations the water molecules are treated classically. The resulting spectra show a blue shift of 0.3 eV in both absorption and emission bands in water compared to the gas phase. A Stokes shift of about 0.1 eV is predicted, and the width of the emission band in solution is significantly broader than the absorption band. These results are consistent with experimental data for BODIPY and closely related dyes, and demonstrate how both absorption and emission spectra in solution can be simulated using a quantum mechanical treatment of the electronic structure of the solute.


Tumor initiating activities of 1-nitropyrene and its nitrated products in SENCAR mice.
The environmental mutagens, 1-nitropyrene, 1,3-dinitropyrene, 1,6-dinitropyrene and 1,8-dinitropyrene were evaluated for their ability to initiate skin tumors in male and female SENCAR mice. 1-Nitropyrene (greater than 99.5% purity) did not induce papilloma formation over a dose range of 0-3.0 mg/mouse after 30 weeks of promotion with 12-O-tetradecanoylphorbol-13-acetate (TPA). Systemic administration of 1-nitropyrene by i.p. injection over a dose range of 0-8.0 mg/mouse followed by topical treatment with TPA also did not induce papilloma formation. A mixture of dinitropyrenes (1,3-dinitropyrene, 1,6-dinitropyrene and 1,8-dinitropyrene (1:1.94:1.95] (99% purity) was dermally applied to SENCAR mice over a dose range of 0-2.0 mg/mouse and the mice subsequently promoted with TPA. A significant induction of papillomas was observed at 30 weeks (0.37-0.39 papillomas/mouse, 26-29% of the mice bearing tumors) at 2.0 mg/mouse. Comparison of this data with that obtained from the same mouse strain using diesel exhaust particulate extracts containing 1-nitropyrene and its nitrated products indicate that these nitrated polynuclear aromatics do not significantly contribute to the mouse skin tumorigenic activity of diesel exhaust particulate extracts.


[Molecular mechanisms of exercise-induced cardiovascular adaptations. Influence of epigenetics, mechanotransduction and free radicals].
The description of mechanisms underlying exercise-induced heart and vascular bed adaptations reveals and highlights the significance of different mechanical and metabolic stimuli that possibly evoke various short-term and long-term regulations and adaptations of these tissues. In this brief review the molecular mechanisms mediated by free radicals and/or mechanical stimulation and, are therefore involved in the modulation of the extracellular matrix and epigenetics-based regulation of the functional genome will be discussed. In the heart and the vascular bed free radicals play important roles in physiological and pathophysiological processes. Exercise leads on the one hand to increased free radicals but on the other hand improves the antioxidative capacity. This phenomenon shifts the cellular oxidative stress balance and also a variety of signal cascades that mediate physiological and pathophysiological heart and vascular bed adaptations. A similar great significance can be attributed to mechanical stimulation which directly or indirectly influences a variety of signaling cascades. It was demonstrated that exercise alters the molecular composition and architecture of the extracellular matrix which in turn plays an important role in the regulation of different mechanical stimuli-mediating signaling cascades. These alterations in the molecular composition and architecture of the extracellular matrix are of high significance for cellular adaptation processes, possibly also in the sense of epigenetic modulations that are actually only indirectly linked to exercise in cardiovascular tissues. However, there is growing evidence that epigenetic modulations mediated by exercise and physical activity can provoke modifications of the functional genome in heart and vascular beds, comparable to already well-described phenomena, e.g. diet or inflammation.


Isolated muscular sarcoidosis presenting as hypercalcaemic renal failure.
The case report describes the details of a man in his 40s admitted for evaluation of renal failure. Biochemical testing revealed parathyroid hormone (PTH) -independent hypercalcaemia. The evaluations for the usual causes such as malignancies, granulomatous diseases, multiple myeloma and vitamin D toxicity were negative. Fluorodeoxyglucose Positron Emission Tomography (FDG-PET) scan identified diffuse uptake in the muscles, and the subsequent muscle biopsy showed non-caseating granulomas suggestive of granulomatous myositis, possibly sarcoidosis, in view of raised ACE levels and the absence of other causes. The patient showed a dramatic response to glucocorticoids, with prompt relief of symptoms and normalisation of serum calcium and creatinine. The case highlights the importance of considering sarcoid myositis in the evaluation of hypercalcaemia and the need for prompt initiation of glucocorticoid therapy to achieve favourable outcomes. The successful use of FDG-PET in diagnosing PTH-independent hypercalcaemia suggests its potential as a valuable tool in the diagnostic algorithm for this condition.


Real-time activity classification in a wearable system prototype for knee health assessment via joint sounds.
An algorithm for performing activity classification for a joint health assessment system using acoustical emissions from the knee is presented. The algorithm was refined based on linear acceleration data from the shank and the thigh sampled at 100 Hz/ch and collected from eight healthy subjects performing unloaded flexion-extension and sit-to-stand motions. The algorithm was implemented on a field-programmable gate array (FPGA)-based processor and has been validated in realtime on a subject performing two minutes of activities consisting of flexion-extension, sit-to-stand, and other motions while standing. When an activity is detected, the algorithm generates an enable signal for high throughput data acquisition of knee joint sounds using two airborne microphones (100 kHz/ch) and two single-axis gyroscope and accelerometer pairs (1 kHz/ch). This approach can facilitate energy-efficient recording of joint sound signatures in the context of flexion-extension and sit-to-stand activities from freely-moving subjects throughout the day, potentially providing a means of evaluating rehabilitation status, for example, following acute knee injury.


Controlled production of stable heterologous proteins in Lactococcus lactis.
The use of Lactococcus lactis (the most extensively characterized lactic acid bacterium) as a delivery organism for heterologous proteins is, in some cases, limited by low production levels and poor-quality products due to surface proteolysis. In this study, we combined in one L. lactis strain use of the nisin-inducible promoter P(nisA) and inactivation of the extracellular housekeeping protease HtrA. The ability of the mutant strain, designated htrA-NZ9000, to produce high levels of stable proteins was confirmed by using the staphylococcal nuclease (Nuc) and the following four heterologous proteins fused or not fused to Nuc that were initially unstable in wild-type L. lactis strains: (i) Staphylococcus hyicus lipase, (ii) the bovine rotavirus antigen nonstructural protein 4, (iii) human papillomavirus antigen E7, and (iv) Brucella abortus antigen L7/L12. In all cases, protein degradation was significantly lower in strain htrA-NZ9000, demonstrating the usefulness of this strain for stable heterologous protein production.


New papulovesicular rash in the course of COVID-19 signaling viral reactivation.
Cutaneous manifestations are considered an infrequent presentation of coronavirus disease 2019 (COVID-19) and are mostly described in outpatient settings. Its onset during the course of the severe COVID-19 disease has been poorly described in severe cases. Studies focused on dermatological manifestations mostly described maculopapular or pernio-like lesions and less frequently vesicular or varicella-like eruption. We described the occurrence of a vesiculopapular eruption in three laboratory-confirmed COVID-19 patients associated with severe lung injury in whom the skin findings preceded viral reactivation and recrudescence of hypoxemia. The potential mechanisms for COVID-19-related cutaneous manifestations include immune hypersensitivity, cytokine-release syndrome, deposition of microthrombi, and vasculitis.


[Anastomoses of the large intestine in standardized diffuse peritonitis in the rat].
A standardized experimental diffuse peritonitis model was developed for the purpose of studying the mechanisms of impaired anastomotic healing in cases of peritonitis. Sham peritonitis or standardized diffuse peritonitis was induced to 42 animals, and anastomosis of the ascending colon was performed 24 hours later. Bursting pressures were measured at six different time intervals, following performance of anastomosis. Significant differences in bursting pressure between sham peritonitis and real peritonitis were restricted to the shortest time interval after performance of anastomosis. More studies will be necessary to investigate the high rates of insufficiency in peritonitis cases.


Hexagonal-close-packed lattice: Ground state and phase transition.
We study the ground state (GS) and the phase transition in a hexagonal-close-packed lattice with both XY and Ising models by using extensive Monte Carlo simulation. We suppose the in-plane interaction J1 and interplane interaction J2, both antiferromagnetic. The system is frustrated with two kinds of GS configuration below and above a critical value of η=J1/J2 (ηc). For the Ising case, one has ηc=0.5 which separates in-plane ferromagnetic and antiferromagnetic states, while for the XY case ηc=1/3 separates the collinear and noncollinear spin configurations. The phase transition is shown to be of first (second) order for η>(<)ηc. The phase diagram in the space (η,T) is shown for both cases.


An aminoacridine derivative for radionuclide therapy: DNA binding properties studied in a novel cell-free in vitro assay.
Radiolabelled DNA-binding compounds can be used to increase the efficiency of radionuclide cancer therapy of disseminated disease. In this work, the aminoacridine compound N-[3-(acridine-9-ylamino)-propyl]-3-iodobenzamide (A3) labelled with the Auger-emitting nuclide 125I using Chloramine-T was studied. Optimal labelling conditions of 125I-A3 were investigated and the interaction with DNA was studied using a novel cell-free in vitro assay with naked human genomic DNA in agarose plugs. This novel assay showed to be simple and reliable. The results verify that 125I-A3 specifically binds DNA with low dissociation and is potent in causing double-strand breaks, yielding 1.0-1.4 breaks per decay. In conclusion, 125I-A3 is a most suitable DNA-binding compound for future therapeutic studies of Auger-electron emitters like 125I.


The role of ipsilateral and contralateral inputs from primary cortex in responses of area 21a neurons in cats.
Neuronal responses in cat visual area 21a were analyzed when the primary visual cortex (areas 17 and 18) was deactivated by cooling. Ipsilateral and contralateral cortices were deactivated separately. Results established that (1) cooling the ipsilateral primary cortex diminished the activity of all area 21a cells and, in 30%, blocked responsiveness altogether, and (2) cooling the contralateral primary cortex initially increased activity in area 21a cells but, with further cooling, reduced it to below the original level although only 9% of cells ceased responding. These findings were then compared to earlier results in which bilateral deactivation of the primary cortex greatly reduced and, in most cases, blocked the activity of area 21a cells (Michalski et al., 1993). Despite the response attenuation following cooling of the primary visual cortex (either ipsilateral or contralateral), neurons of area 21a retained their original orientation specificity and sharpness of tuning (measured as the half-width at half-height of the orientation tuning curve). Direction selectivity also tended to remain unchanged. We concluded that for area 21a cells (1) the ipsilateral primary cortex provides the main excitatory input; (2) the contralateral primary cortex supplies a large inhibitory input; and (3) the nature of orientation specificity, sharpness of orientation tuning, and direction selectivity are largely unaffected by removal of the ipsilateral hemisphere excitatory input or the contralateral hemisphere inhibitory input.


In vivo and ex vivo cetuximab sensitivity assay using three-dimensional primary culture system to stratify KRAS mutant colorectal cancer.
In clinic, cetuximab, an anti-EGFR antibody, improves treatment outcomes in colorectal cancer (CRC). KRAS-mutant CRC is generally resistant to cetuximab, although difference of the sensitivity among KRAS-mutants has not been studied in detail. We previously developed the cancer tissue-originated spheroid (CTOS) method, a primary culture method for cancer cells. We applied CTOS method to investigate whether ex vivo cetuximab sensitivity assays reflect the difference in sensitivity in the xenografts. Firstly, in vivo cetuximab treatment was performed with xenografts derived from 10 CTOS lines (3 KRAS-wildtype and 7 KRAS mutants). All two CTOS lines which exhibited tumor regression were KRAS-wildtype, meanwhile all KRAS-mutant CTOS lines grew more than the initial size: were resistant to cetuximab according to the clinical evaluation criteria, although the sensitivity was quite diverse. We divided KRAS-mutants into two groups; partially responsive group in which cetuximab had a substantial growth inhibitory effect, and resistant group which exhibited no effect. The ex vivo signaling assay with EGF stimulation revealed that the partially responsive group, but not the resistant group, exhibited suppressed ERK phosphorylation ex vivo. Furthermore, two lines from the partially responsive group, but none of the lines in the resistant group, exhibited a combinatory effect of cetuximab and trametinib, a MEK inhibitor, ex vivo and in vivo. Taken together, the results indicate that ex vivo signaling assay reflects the difference in sensitivity in vivo and stratifies KRAS mutant CTOS lines by sensitivity. Therefore, coupling the in vivo and ex vivo assays with CTOS can be a useful platform for understanding the mechanism of diversity in drug sensitivity.


The relationship between the Five-Factor Model personality traits and peptic ulcer disease in a large population-based adult sample.
The current study examined the relationship between the Five-Factor Model personality traits and physician-confirmed peptic ulcer disease (PUD) diagnosis in a large population-based adult sample, controlling for the relevant behavioral and sociodemographic factors. Personality traits were assessed by participants themselves and by knowledgeable informants using the NEO Personality Inventory-3 (NEO PI-3). When controlling for age, sex, education, and cigarette smoking, only one of the five NEO PI-3 domain scales - higher Neuroticism - and two facet scales - lower A1: Trust and higher C1: Competence - made a small, yet significant contribution (p < 0.01) to predicting PUD in logistic regression analyses. In the light of these relatively modest associations, our findings imply that it is certain behavior (such as smoking) and sociodemographic variables (such as age, gender, and education) rather than personality traits that are associated with the diagnosis of PUD at a particular point in time. Further prospective studies with a longitudinal design and multiple assessments would be needed to fully understand if the FFM personality traits serve as risk factors for the development of PUD.


Surface-Induced 2D/1D Heterostructured Growth of ReS2/CoS2 for High-Performance Electrocatalysts.
Two-dimensional/one-dimensional (2D/1D) heterostructures have received much attention from researchers for their abundant catalytically active sites and low contact resistance due to formation of chemical bonds at the interface. The investigation of such heterostructures, however, is confined to lattice-matched materials, which severely limits the material candidates. Herein, we demonstrate a lattice-mismatched 2D/1D heterostructured electrocatalyst consisting of 2D ReS2 nanosheets and 1D CoS2 nanowires. We propose that the higher surface energy of the CoS2 nanowire and the lattice mismatch between 1D and 2D units are crucial for the growth process of ReS2 nanosheets. More importantly, the terminal S2- exposed on the surface of CoS2 nanowires serves not only as the nucleus of ReS2 nanosheets but also as a bridge to enhance electron transport efficiency. Thus, the ReS2/CoS2 heterostructures show outstanding hydrogen evolution reaction performance. This work is of general interest for the design of complex multidimensional nano-heterostructures with outstanding functionalities.


Identification of Candidate Genes Involved in Renal Ischemia/Reperfusion Injury.
Renal ischemia/reperfusion injury (IRI) is a main risk factor for the occurrence of delayed graft function or primary graft nonfunction of kidney transplantation. However, it lacks ideal molecular markers for indicating IRI in kidney transplantation. The present study is to explore novel candidate genes involved in renal IRI. Experimental renal IRI mouse models were constructed, and the differentially expressed genes were screened using a microarray assay. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were performed. The expression of genes was detected using real-time qPCR assay. Western blotting and immunohistochemistry staining assays were performed for protein determination. We identified that renal IRI induced the upregulation of SPRR2F, SPRR1A, MMP-10, and long noncoding RNA (lncRNA) Malat1 in kidney tissues for 479.3-, 4.98-, 238.1-, and 3.79-fold, respectively. The expression of miR-139-5p in kidney tissues of IRI-treated mice was decreased to 40.4% compared with the sham-operated mice. These genes are associated with keratinocyte differentiation, regeneration and repair of kidney tissues, extracellular matrix degradation and remodeling, inflammation, and cell proliferation in renal IRI. Identification of novel biomarkers involved in renal IRI may provide evidences for the diagnosis and treatment of renal IRI.


Targeting cancer stem cells in glioblastoma multiforme using mTOR inhibitors and the differentiating agent all-trans retinoic acid.
Glioblastoma multiforme (GBM), the most aggressive primary brain tumor, portends a poor prognosis despite current treatment modalities. Recurrence of tumor growth is attributed to the presence of treatment-resistant cancer stem cells (CSCs). The targeting of these CSCs is therefore essential in the treatment of this disease. Mechanistic target of rapamycin (mTOR) forms two multiprotein complexes, mTORC1 and mTORC2, which regulate proliferation and migration, respectively. Aberrant function of mTOR has been shown to be present in GBM CSCs. All-trans retinoic acid (ATRA), a derivative of retinol, causes differentiation of CSCs as well as normal neural progenitor cells. The purpose of this investigation was to delineate the role of mTOR in CSC maintenance, and to establish the mechanism of targeting GBM CSCs using differentiating agents along with inhibitors of the mTOR pathways. The results demonstrated that ATRA caused differentiation of CSCs, as demonstrated by the loss of the stem cell marker Nestin. These observations were confirmed by western blotting, which demonstrated a time-dependent decrease in Nestin expression following ATRA treatment. This effect occurred despite combination with mTOR (rapamycin), PI3K (LY294002) and MEK1/2 (U0126) inhibitors. Expression of activated extracellular signal-regulated kinase 1/2 (pERK1/2) was enhanced following treatment with ATRA, independent of mTOR pathway inhibitors. Proliferation of CSCs, determined by neurosphere diameter, was decreased following treatment with ATRA alone and in combination with rapamycin. The motility of GBM cells was mitigated by treatment with ATRA, rapamycin and LY29002 alone. However, combination treatment augmented the inhibitory effect on migration suggesting synergism. These findings indicate that ATRA-induced differentiation is mediated via the ERK1/2 pathway, and underscores the significance of including differentiating agents along with inhibitors of mTOR pathways in the treatment of GBM.


[Treatment of patients with resected lower jaw with removable overdentures].
An orthopedic treatment mode is described of patients with hemiresection of the lower jaw with the retention of several teeth on the residual fragment. A postresectional overdenture with telescopic fixation and leading buccal phalanges have been worked out upon the properly formed, capped and splinted supporting teeth. The good results from the treatment reveal that overdentures are indicated for maxillofacial prosthesis.


Ex Vivo Treatment with a Polyphenol-Enriched Cocoa Extract Ameliorates Myocardial Infarct and Postischemic Mitochondrial Injury in Normotensive and Hypertensive Rats.
Our objective was to determine the effects of a polyphenol-enriched cocoa extract (PCE) on myocardial postischemic alterations in normotensive (Wistar rats, W) and spontaneously hypertensive rats (SHR). Isolated hearts were submitted to 110 min of perfusion or 20 min stabilization, 30 min global ischemia, and 60 min reperfusion (R). Other hearts were treated with PCE at the onset of R. Infarct size, the reduced glutathione (GSH), and the expression of phospho-Akt, P-GSK-3β, and P-eNOS were assessed. In isolated mitochondria, the Ca(2+)-mediated response of mitochondrial permeability transition pore (mPTP), membrane potential (Δψm), and superoxide production were determined. PCE decreased infarct size, partly preserved GSH, increased the P-Akt, P-GSK-3β, and P-eNOS contents, improved mPTP response to Ca(2+), decreased the superoxide production, and restored Δψm. These data show that PCE decreases the cardiac postischemic damage in W rats and SHR and suggest that Akt/GSK-3β/eNOS dependent pathways are involved.


Modifications and evaluation of double antibody radioimmunoassay of human carcinoembryonic antigen.
Double antibody radioimmunoassay of carcinoembryonic antigen (CEA), a cancer-associated antigen of the human digestive system, was subjected to certain modifications and critically evaluated. Modifications pertained to: (a) the production of a high titer goat anti-CEA antiserum that was rendered highly specific by solid phase immunoabsorption with cyanogen bromide-activated Sepharose conjugates of normal plasma liver, and colon perchloric acid-soluble glycoprotein antigens: (b) the introduction of suitable alterations in the experimental conditions of radioiodination procedure to minimize and to prevent breakdown of the antigen, thus prolonging the storage of the labeled antigen; (c) the extended incubation period of CEA-anti-CEA immune reaction; and (d) the use of sodium acetate buffer, pH 6.1. Furthermore, the use of an automatic pipetting station for accurate and rapid reagent dispensation and statistical analysis of the radioimmunoassay data on a modern computer to ensure strict quality control of the assay provided some definite improvement over the existing assay.


Analysis of Fatty Acid Metabolism in Fetal and Failing Hearts by Single-Cell RNA Sequencing Revealed SLC27A6 as a Critical Gene in Heart Maturation.
Heart failure is associated with shifts in substrate preferences and energy insufficiency. Although cardiac metabolism has been explored at the organ level, the metabolic changes at the individual cell level remain unclear. This study employed single-cell ribonucleic acid (RNA) sequencing to investigate the cell-type-specific characteristics of gene expression related to fatty acid metabolism. Single-cell RNA sequencing data from fetal hearts were processed to analyze gene expression patterns related to fatty acid metabolism. Immunofluorescence staining and Western blotting techniques were employed to validate the expression of specific proteins. Additionally, calcium recording and contractility measurements were performed to assess the functional implications of fatty acid metabolism in cardiomyocytes. Based on single-cell RNA sequencing data analysis, we found that a decrease in overall energy requirements underlies the downregulation of fatty acid oxidation-related genes in the later period of heart maturation and the compensatory increase of fatty acid metabolism in individual cardiomyocytes during heart failure. Furthermore, we found that solute carrier family 27 member 6 (SLC27A6), a fatty acid transport protein, is involved in cardiac maturation. SLC27A6 knockdown in human induced pluripotent stem cell-derived cardiomyocytes resulted in an immature cardiomyocyte transcriptional profile, abnormal morphology, impaired Ca2+ handling activity, and contractility. Overall, our study offers a novel perspective for exploring cardiac fatty acid metabolism in fetal and failing hearts along with new insights into the cellular mechanism underlying fatty acid metabolic alterations in individual cardiac cells. It thus facilitates further exploration of cardiac physiology and pathology.


Complementary Adsorption of Binary Levelers for Highly Uniform Cu Micropillars.
Highly uniform Cu micropillars were electrodeposited on a Cu seed layer in a micropatterned photoresist in the presence of binary levelers containing pyrrolidine and pyridine functional groups. The adsorption behaviors of binary levelers, i.e., 3-hydroxy-6-(tert-butyl)pyridine and pyridinium tribromide in an electroplating bath were investigated by chronopotentiometry with sequential injection of levelers and linear sweep voltammetry with a rotating disk electrode. From electrochemical analysis, pyridinium tribromide composed of a positively charged pyridine ring was strongly adsorbed on the Cu seed surface, relative to 3-hydroxy-6-(tert-butyl)pyridine. Additional microscopy, surface roughness, and nitrogen concentration analyses revealed that the binary levelers were preferentially adsorbed on the center and the edge region of the Cu seed, resulting in a uniform Cu pillar profile. The possible mechanism of highly uniform Cu pillar deposition was discussed in terms of the adsorption behaviors of the levelers dependent on their molecular structures.


Microsporidiosis in a Gouldian finch (Erythrura [Chloebia] gouldiae).
A 12-day-old nestling Gouldian finch (Erythrura [Chloebia] gouldiae) was presented for investigation of a mortality problem in nestling finches raised by Bengalese finch foster parents. On histological examination, large numbers of spores consistent with a microsporidian organism were present within the small intestinal mucosa. Electron microscopy and molecular studies (sequencing the 5' end of the ssu rRNA gene) further defined the organism as Encephalitozoon hellem. Sequence homology with other eukaryotes was determined using a BLASTN search from the NCBI GenBank database. The finch isolate sequences showed greater than 99% homology with those of previously reported human and avian isolates.


Evaluation of thermotolerant yeasts in controlled simultaneous saccharifications and fermentations of cellulose to ethanol.
Simultaneous saccharification and fermentation (SSF) experiments were performed at selected temperatures (37, 41, and 43 degrees C) to obtain comprehensive material balance and performance data for several promising strains of thermotolerant yeast. Parameters measured were ethanol concentration, yeast cell density, and residual sugar and cellulose concentrations. The three yeasts Saccharomyces uvarum, Candida brassicae, and C. lusitaniae and two mixed cultures of Brettanomyces clausenii with S. cerevisiae (mixed culture I) and C. Iusitaniae with S. uvarum (mixed culture II) exhibited rapid rates of fermentation, high ethanol yields, strong viability, or high cellobiase activity. Overall, mixed culture II at 41 degrees C performed better than either component yeast by themselves because it combined a cellobiose fermenting capability with the high ethanol tolerance and rapid glucose fermentation of conventional industrial yeasts. Thus, the mixed cultures provide good initial rates by preventing buildup of cellobiose (a strong inhibitor of enzyme activity) while attaining high ultimate yields of ethanol for high cellulase concentrations. However, C. brassicae and S. uvarum gave similar results to mixed culture II at 37 degrees C.


Recurrent focal segmental glomerulosclerosis in renal allograft recipients: role of human leukocyte antigen mismatching and other clinical variables.
Recurrence of focal segmental glomerulosclerosis (FSGS) after renal transplantation impacts long-term graft survival and limits access to transplantation. We hypothesized that HLA donor/recipient matching could be used as a surrogate marker of recurrence. In a retrospective study of 42 pediatric and 77 adult subjects with primary FSGS, transplanted from 1990 to 2007 at a single center, we analyzed the degree of donor/recipient HLA compatibility and other clinical variables associated with FSGS recurrence. There were total of 131 allografts for primary FSGS (11 subjects were transplanted twice, and 1 had a third allograft) with 20 cases of FSGS recurrence (17 children) in the primary allograft, and two children who had FSGS recurrence in the second allograft. Fifty-two subjects (40%) were African American, and 66 (50%) Caucasians. Recurrent FSGS and controls were not different for age at transplant, gender, donor source, acute/chronic rejection episodes, and HLA matches. Recurrent FSGS was not associated with HLA mismatches; power equals 83%. Immunosuppressive regimen had no effect on recurrence of FSGS, P = .75. Recurrent FSGS is not associated with HLA mismatching, acute cellular or vascular rejection, and occurs primarily in the pediatric population.


Regulation of AMP-activated protein kinase activity by G-protein coupled receptors: potential utility in treatment of diabetes and heart disease.
G-protein coupled receptors (GPCRs) comprise the largest and most diverse family of membrane receptors in the human genome, relaying information from a vast array of external stimuli. GPCRs are targets for approximately 30% of all current therapeutic agents. Recently some GPCRs have been shown to mediate part of their effects through activation of AMP-activated protein kinase (AMPK), a sensor of whole body energy status that plays a pivotal role in whole body energy balance by integrating signals in the periphery and central nervous system. It regulates glucose and lipid metabolism, food intake and body weight, making it an attractive target for the treatment of diseases such as type 2 diabetes and obesity. It mediates the effects of several important adipokines such as leptin and adiponectin and is thought to be responsible for the antidiabetic effects of metformin and thiazolidinediones. A diverse number of GPCRs (including adrenoceptors, cannabinoid receptors, ghrelin receptors, melanocortin receptors) modulate AMPK activity. This review focuses on the regulation of AMPK by GPCRs and signaling intermediates of GPCR signaling such as cyclic AMP and calcium, and how GPCR signaling can modulate AMPK activity by several different mechanisms, and the therapeutic implications of AMPK activation by GPCRs.


Geographical Environment Factors and Risk Assessment of Tick-Borne Encephalitis in Hulunbuir, Northeastern China.
Tick-borne encephalitis (TBE) is one of natural foci diseases transmitted by ticks. Its distribution and transmission are closely related to geographic and environmental factors. Identification of environmental determinates of TBE is of great importance to understanding the general distribution of existing and potential TBE natural foci. Hulunbuir, one of the most severe endemic areas of the disease, is selected as the study area. Statistical analysis, global and local spatial autocorrelation analysis, and regression methods were applied to detect the spatiotemporal characteristics, compare the impact degree of associated factors, and model the risk distribution using the heterogeneity. The statistical analysis of gridded geographic and environmental factors and TBE incidence show that the TBE patients mainly occurred during spring and summer and that there is a significant positive spatial autocorrelation between the distribution of TBE cases and environmental characteristics. The impact degree of these factors on TBE risks has the following descending order: temperature, relative humidity, vegetation coverage, precipitation and topography. A high-risk area with a triangle shape was determined in the central part of Hulunbuir; the low-risk area is located in the two belts next to the outside edge of the central triangle. The TBE risk distribution revealed that the impact of the geographic factors changed depending on the heterogeneity.