Airway smooth muscle cell tone amplifies contractile function in the presence of chronic cyclic strain.
Chronic contractile activation, or tone, in asthma coupled with continuous stretching due to breathing may be involved in altering the contractile function of airway smooth muscle (ASM). Previously, we (11) showed that cytoskeletal remodeling and stiffening responses to acute (2 h) localized stresses were modulated by the level of contractile activation of ASM. Here, we investigated if altered contractility in response to chronic mechanical strain was dependent on repeated modulation of contractile tone. Cultured human ASM cells received 5% cyclic (0.3 Hz), predominantly uniaxial strain for 5 days, with once-daily dosing of either sham, forskolin, carbachol, or histamine to alter tone. Stiffness, contractility (KCl), and "relaxability" (forskolin) were then measured as was cell alignment, myosin light-chain phosphorylation (pMLC), and myosin light-chain kinase (MLCK) content. Cells became aligned and baseline stiffness increased with strain, but repeated lowering of tone inhibited both effects (P < 0.05). Strain also reversed a negative tone-modulation dependence of MLCK, observed in static conditions in agreement with previous reports, with strain and tone together increasing both MLCK and pMLC. Furthermore, contractility increased 176% (SE 59) with repeated tone elevation. These findings indicate that with strain, and not without, repeated tone elevation promoted contractile function through changes in cytoskeletal organization and increased contractile protein. The ability of repeated contractile activation to increase contractility, but only with mechanical stretching, suggests a novel mechanism for increased ASM contractility in asthma and for the role of continuous bronchodilator and corticosteroid therapy in reversing airway hyperresponsiveness.


Where to from here? Posthumous healthcare data, digital e(lectronic)-mortality and New Zealand's healthcare future.
Ongoing improvements in digital data acquisition and storage has led to the phenomenon of e(lectronic)-mortality, where digital data can now exist for a potentially infinite period. Globally, many countries are facilitating the acquisition and researcher-led access to large-scale, population-based digitised healthcare data sets. Their utilisation has led to numerous positive advances in healthcare. New Zealand's medical record system is becoming increasingly digitised, and as a consequence there will be an ever-increasing resource of posthumous healthcare data stored digitally, including genomic information. Such data could be utilised for research purposes, and developing such a consolidated resource could improve healthcare outcomes in our own nation and allow us to parallel global progress in healthcare research trends. This viewpoint article explores the issues surrounding, and potential for utilisation of, a national resource of posthumous digital healthcare data. Currently, there appear to be no legal barriers to the large-scale acquisition and utilisation of posthumous healthcare data in New Zealand, however, previous legislation may not have been developed with developments in technology or e-mortality in mind. Ethically, culturally and socially there are many challenges to address, including issues surrounding obtaining consent, respecting privacy, management of incidental findings, maintaining anonymity and ensuring community support for such a resource. Despite the potential for widespread health benefits that utilisation of posthumous healthcare data in this country may facilitate, wide and ongoing consultation is required to examine how such a precious resource can be enabled for the downstream benefit of all New Zealanders.


Hsa-miR-1269a up-regulation fosters the malignant progression of esophageal squamous cell carcinoma via targeting FAM46C.
Esophageal squamous cell carcinoma (ESCC) is a malignancy of the alimentary tract resulting in death worldwide. The role and underlying mechanism of hsa-miR-1269a in the progression of ESCC remain unclear. In this study, hsa-miR-1269a was screened by differential expression analysis in TCGA, and its target gene FAM46C was predicted. qRT-PCR was conducted to assay the expression of hsa-miR-1269a and FAM46C in ESCC cells. The results showed that hsa-miR-1269a was upregulated in ESCC tissues and cell lines. Hsa-miR-1269a overexpression stimulated the proliferation, migration, and invasion capacities of ESCC cells, and FAM46C overexpression inhibited these phenotypes. Dual-luciferase assay verified that hsa-miR-1269a could target FAM46C. Next, qRT-PCR and western blot demonstrated that hsa-miR-1269a overexpression downregulated FAM46C. Rescue experiments revealed that hsa-miR-1269a accelerated the malignant progression of ESCC through FAM46C down-regulation. These results indicate that the interaction between hsa-miR-1269a and FAM46C plays a regulatory role in driving the malignant progression of ESCC cells, thereby providing a novel molecular mechanism for understanding ESCC.


Delayed leukoencephalopathy in survivors with small cell lung cancer.
Six patients with small cell lung cancer developed a slowly progressive neurologic syndrome characterized by apathy, abulia, memory loss, gait ataxia, and corticospinal tract signs 26 to 50 months (mean, 35.2 months) after prophylactic cranial irradiation and systemic chemotherapy. In each case this was accompanied by CT and/or MRI evidence of changes in the periventricular white matter. These patients are long-term survivors (41 to 69 months) and do not have CNS metastases.


The relationship of histologic activity to serum ALT, HCV genotype and HCV RNA titers in chronic hepatitis C.
It is unclear whether serum ALT levels or virological characteristics of hepatitis C virus(HCV) including HCV genotypes and HCV RNA titers, can reflect the degree of histological injury in chronic hepatitis C. The aim of this study was to investigate the relationships between the levels of histological damage and serum ALT levels, HCV genotypes or circulating HCV RNA titers in chronic hepatitis C. A total of 56 patients underwent liver biopsy and the histological activity index (HAI) was evaluated by Knodell's scoring system. HCV genotype by RT-nested PCR and HCV RNA quantitation by competitive RT-PCR were performed. Thirty-four patients were infected with HCV genotype 1b, 20 patients with genotype 2a, and 2 patients with undetermined type. Serum ALT levels were not positively correlated with total HAI score or HCV RNA titers, but showed a linear correlation with scores of piecemeal necrosis (r=0.32, p<0.05) and portal inflammation (r=0.27, p<0.05). HCV genotype had no significant correlation with RNA titers, HAI score or with serum ALT levels. Also, no statistical relationship was seen between HCV RNA titer and HAI score. These results suggest that liver histology is essential to evaluate the severity of chronic hepatitis C precisely.


Analysis of pulse front tilt in simultaneous spatial and temporal focusing.
Simultaneous spatial and temporal focusing (SSTF) has gained enormous interest for controlling the focal region of ultrashort pulses in the material processing community. In this paper, we provide theoretical insight in the nature of SSTF. We use numerical simulations to propagate the initial pulse through the focusing lens and into the focal region. By that we can investigate the appearance of the pulse front tilt (PFT), which has been experimentally obtained in SSTF. This is further deepened by a mathematical investigation which shows that PFT is a fundamental consequence of SSTF. Next we follow the idea to use an initial PFT in order to influence or even compensate the PFT in the focal region. Again that is done by simulations as well as mathematical investigations. We found that an initial PFT cannot significantly influence the PFT in the focal region.


Staphylococcus aureus Bacteremia Complicated by Psoas Abscess and Infective Endocarditis in a Patient with Atopic Dermatitis.
The close relationship between atopic dermatitis (AD) and infective endocarditis (IE) has been implicated. Staphylococcus aureus colonization is frequently seen observed in AD patients' skin lesions. Although a case of IE due to S. aureus bacteremia in an AD patient has been sporadically reported, a case of S. aureus bacteremia complicated by psoas abscess and IE has not been previously reported. A 42-year-old man with a history of AD presented to our hospital complaining of fever, fatigue, chills, lower right back pain, and poor appetite for a week. His blood cultures showed growth of S. aureus. On day 3, the patient presented acute cardiac failure and was diagnosed with IE based on echocardiogram examination. Since the patient's cardiac failure did not respond to medication, an emergency surgery was performed on the fourth day of hospitalization. The patient underwent successful surgical treatment of the heart lesions and subsequent percutaneous drainage of psoas abscess and received intensive antibiotics, which successfully improved his condition. Our report emphasizes awareness of the association between AD and invasive S. aureus infections.


Effects of corticotropin-releasing hormone on luteinizing hormone, testosterone, and cortisol secretion in intact male rhesus macaques.
We previously have shown that 6 h of restraint stress in intact male rhesus macaques (Macaca mulatta) suppresses plasma levels of both LH and testosterone and that this effect lasts beyond the period of restraint. Since corticotropin-releasing hormone (CRH) inhibits both the GnRH pulse generator and LH release in ovariectomized macaques and is generally thought to be the central mediator of stress-induced inhibition of gonadotropin release, we investigated the influence of CRH administration on LH and testosterone in unrestrained intact male rhesus macaques. Blood samples were collected from 5 intact male macaques at 15-min intervals for 15 h from a remote site. During this time, each animal received a 4-h infusion of CRH (100-micrograms bolus followed by 100 micrograms/h for 4 h) through an indwelling jugular catheter. Blood samples were collected for an additional 8 h after cessation of the CRH infusion. ACTH and cortisol levels were significantly elevated during and after the CRH infusion and were comparable to levels observed during restraint. Although LH levels appeared lower in animals given CRH, they were not different from those in untreated control males. In some animals, CRH appeared to stimulate or prolong LH release. Testosterone levels in CRH-treated animals were significantly lower than in controls both during and after CRH administration. In some instances, increases in plasma LH were not accompanied by a rise in testosterone. This would suggest that as was observed in restrained animals, stress exerts either a direct or an indirect inhibition of testicular testosterone secretion. The present data indicate that this effect may be mediated by CRH.


Jackfruit Anaphylaxis Due to Cross Reactivity with Latex.
Growing evidence has demonstrated that many common plant foods such as mangos, kiwis and jackfruit lead to cross reactivity with the latex antigen in latex allergic patients. Here, we discuss the case of a 68 year old female of Bangladeshi descent who developed shortness of breath in the setting of anaphylaxis following the ingestion of jackfruit (Artocarpus heterophyllus). The patient had a history of latex allergy described as mild rash along with seasonal allergies causing mild rhinorrhea, congestion and sneezing. Given the strong cultural significance of jackfruit consumption in Asian countries and growing popularity in the use of jackfruit as a superfood meat alternative in Western nations, along with growing Asian population, there is a need for extensive education on the cross reactivity between plant foods and latex to prevent deadly cross-reactivity induced anaphylaxis. With this case report, we hope to raise awareness of this rare, yet morbid association.


Trends in Sunscreen Use Among US Middle and High School Students, 2007-2019.
The purpose of this project is to analyze trends in sunscreen usage among middle and high school students from the Centers of Disease Control's National Youth Risk Behavior Surveys (YRBS) from the years 2007 to 2019. Data from the 2007-2019 National YRBS were analyzed. YRBS is a cross-sectional survey of health risk behaviors among middle and high school students (grades 6-12) in the United States. Students were asked questions regarding sunscreen usage and demographic information, including age, race/ethnicity, and gender. From 2007 to 2019, the average mean sunscreen usage for all students increased by 4% between every consecutive year studied. Mean sunscreen usage among all racial groups studied, other than Native Hawaiian or Other Pacific Islanders, increased. Females were 92.7% more likely to use sunscreen than males each year. The mean sunscreen usage decreased by 5% with an increase in a student's age. Though overall sunscreen use in adolescents increased over the time period from 2007 to 2019, sunscreen use still remains limited due to a variety of factors possibly including cost, tanning, and different socio-cultural perceptions of sunscreen.


Application of Bottlebrush Block Copolymers as Photonic Crystals.
Brush block copolymers are a class of comb polymers that feature polymeric side chains densely grafted to a linear backbone. These polymers display interesting properties due to their dense functionality, low entanglement, and ability to rapidly self-assemble to highly ordered nanostructures. The ability to prepare brush polymers with precise structures has been enabled by advancements in controlled polymerization techniques. This Feature Article highlights the development of brush block copolymers as photonic crystals that can reflect visible to near-infrared wavelengths of light. Fabrication of these materials relies on polymer self-assembly processes to achieve nanoscale ordering, which allows for the rapid preparation of photonic crystals from common organic chemical feedstocks. The characteristic physical properties of brush block copolymers are discussed, along with methods for their preparation. Strategies to induce self-assembly at ambient temperatures and the use of blending techniques to tune photonic properties are emphasized.


Diabetes self-care activities: a community-based survey in urban southern India.
Diabetes is a lifestyle disease and can be successfully managed by good self-care activities such as diet, exercise, monitoring and drug adherence. Adequate baseline information about the prevalence of good self-care activities is not available from India. We aimed to estimate the existing self-care behaviours and factors influencing these behaviours among adult patients with type 2 diabetes in urban southern India. A cross-sectional survey was conducted using a cluster design in an urban community in southern India. The Summary Diabetes Self-Care Activities questionnaire was used to collect information on diet, exercise, monitoring of blood sugars and adherence to drugs. Risk factors such as marital status, socioeconomic status, depression, benefit-finding and duration of illness, which are likely to influence self-care behaviour, were assessed. Good dietary behaviour was present in 29% (95% CI 20.8%-37.2%), good exercise behaviour in 19.5% (95% CI 17.4%-21.6%), regular blood sugar monitoring in 70% (95% CI 62.2%-77.8%) and drug adherence in 79.8% (95% CI 75.1%-84.5%). Being male (OR 3.38; 95% CI 1.541-7.407) and married (OR 5.60; 95% CI 1.242-25.212) significantly favoured good exercise behaviour. Being married (OR 2.322; 95% CI 1.104-4.883) and belonging to the higher socioeconomic status (OR 2.713; 95% CI 1.419-5.190) were significantly associated with monitoring of blood sugars. Self-care activities with respect to diet and exercise are poor in the population studied. The self-care activities relating to blood sugar monitoring and drug adherence are good. Improving self-care behaviour among patients with diabetes in India should start with adequate targeted health education.


The Caudal Skeleton of the Zebrafish, Danio rerio, from a Phylogenetic Perspective: A Polyural Interpretation of Homologous Structures.
The structure of the caudal skeleton of extant teleost fishes has been interpreted in two different ways. In a diural interpretation, a caudal skeleton is composed of two centra articulated with one to six hypurals. Most subsequent authors have followed this interpretation. In contrast, a polyural interpretation considers the teleost fin to be derived from a fully metameristic ancestral bauplan originally composed of a one-to-one relationship between neural arches, centra (when present), and hypurals. Three different interpretations of the identity and homology of skeletal components of the caudal skeleton of the teleost fish Danio rerio have been proposed, two from a diural perspective and one from a polyural perspective. We examine each caudal skeletal component of Danio rerio from both a developmental and phylogenetic perspective. We propose that a polyural interpretation of structures is consistent with the current interpretation of the basal neopterygian caudal fin for this model organism rather than the older diural interpretation that does not take into account the metamerism observed in caudal structures during development. The polyural interpretation suggests several shared evolutionary innovations of major clades that would remain undiscovered under the older diural naming paradigm and makes the terminology of the parts of the caudal fin of Danio rerio strictly comparable to more basal fishes.


Continuity of care in a university-based practice.
Effects of changes in a pediatric practice--expansion of the number of pediatricians and incorporation into a university hospital setting--on continuity of care and utilization were examined by means of a longitudinal study of a sample of 63 families. Continuity of care was measured by the following index: the number of visits with own physician divided by the total number of pediatric visits per year. Although continuity of well-child visits remained unchanged at the university setting, the continuity of sick visits declined markedly. An increased use of doctor visits for illness care was observed; its relationship with the decline in continuity is analyzed and discussed. While continuity is inherent in a small partnership practice, it is not so in a larger medical organization, particularly when involvement in patient care is part time. In such an organization, deliberate arrangements that enable patients with acute needs to receive care from their own doctors are needed.


Granulocyte migration abnormality in patients suffering from seasonal allergic rhinitis: failure of treatment with cimetidine.
Granulocyte locomotory responses in 5 patients with symptomatic seasonal allergic rhinitis were lower compared with similar responses from 27 normal nonallergic controls. In a subsequent controlled, double-blind crossover study, neither cimetidine (histamine H2-receptor blocker) nor placebo improved these responses. In our in vitro study, histamine did not inhibit granulocyte responses to chemotactic attractant. These results indicate that defective granulocyte response in patients with seasonal allergic rhinitis may be due to factors other than or in addition to histamine.


Problem-based, peer-facilitated education about antibiotic prescribing.
To assess the feasibility and acceptability of a problem-based, peer-facilitated educational workshop about antibiotic prescribingfor GPs. All 39 GPs working in an average sized Local Health Care Co-operative (LHCC) in Glasgow. Prospective collection of information about 10 prescriptions for antibiotics to assess learning needs in relation to antibiotic prescribing. Two and a half hour workshop involving problem-based group work based on the needs assessment and discussions with a consultant microbiologist, prescribing adviser and academic GP. Written feedback about the process of the educational intervention immediately after the workshop and outcome feedback collected after four weeks. Telephone interviews with non-participants. Twenty-four GPs agreed to participate in the study. 19 of these completed the Needs Assessment and 14 of these completed the workshop. The method of learning needs assessment and the educational workshop were highly acceptable. "No time, too busy" was the main reason given for not taking part in the study. Some GPs are unable to participate as fully as they would like in continuing professional learning activities due to competing workload pressures. Further research is required to investigate the necessary type and balance of learning activities and the barriers to engagement to ensure the most effective use of clinician time available for continuing educational activities.


Inhibitory effect of Mao-Bushi-Saishin-to on prostaglandin E2 synthesis in C6 rat glioma cells.
Effect of Mao-Bushi-Saishin-to (Ma-Huang-Fu-Zi-Xi-Xin-Tang: MBS) on prostaglandin E(2) (PGE(2)) production was investigated using C6 rat glioma cells. Mao or Saishin inhibited histamine-induced PGE(2) production while MBS slightly decreased and Bushi increased it. MBS and Mao inhibited and Bushi enhanced A23187-induced PGE(2) production while Saishin had no effect. Concomitantly, Mao inhibted, but Bushi fascilitated, histamine- and A23187-induced phosphorylation of extracellular signal-regulated kinase (ERK)1/2. Treatment of MBS, Mao and also Saishin increased cAMP content. From these results, MBS inhibit PGE(2) production in C6 cells, mainly due to Mao but also due to Saishin at least in part, and the counteraction of Bushi. The former effect is mediated by formation of cAMP and resulting inhibition of ERK1/2-phosphorylation.


Reduced Basis Set Dependence in Anharmonic Frequency Calculations Involving Localized Coordinates.
Localized normal coordinates are known to be effective in speeding up anharmonic frequency calculations by reducing the complexity of the nuclear Hamiltonian and wave function. Displacing atoms in localized coordinates can also cause relatively small changes in the electronic structure, which can be exploited for further computational efficiency improvements during ab initio electronic structure calculations of the potential energy surface by reducing the electronic basis set dependence. Three different schemes for reducing the basis set dependence have been investigated in this work. These include combining localized coordinate schemes with general mixed basis sets, distance based force-field reductions, and using coordinate specific basis sets. The importance of accurately describing electronic interactions is found to diminish both for multicoordinate terms involving the displacement of remote atoms and when describing the interactions between more remote atoms within specific coordinates.


Screening and brief interventions for alcohol: attitudes, knowledge and experience of community pharmacists in Auckland, New Zealand.
Screening and brief intervention (SBI) for alcohol consumption in primary care have been shown to be effective in reducing drinking. This role has not been taken up by community pharmacists. This study aimed to explore attitudes, knowledge, barriers and incentives towards involvement of community pharmacists in New Zealand with problem drinkers. A postal survey (three mailshots) of community pharmacies was undertaken in Auckland, New Zealand, followed by a 10% random sample non-responder-follow-up. A response rate of 39.1% to the postal survey was obtained. In general, knowledge of alcohol content of drinks and recommended safe drinking limits was poor. Respondents were generally well motivated towards undertaking this role, but lacked knowledge, skills and confidence. On follow-up, non-responders were found to be less knowledgeable, but had similar attitudes to respondents, indicating potential for extrapolation with regard to beliefs and views on this subject. This novel study has found that there is potential for involvement of community pharmacists in New Zealand in SBI for problem drinkers. Lack of knowledge can be remedied relatively easily; greater hurdles such as lack of skills, and lack of confidence may be offset by the high level of motivation of this group. This is an area where community pharmacists can further their professional scope of practice by being in an essential position to provide this service.


[Aortic involvement in patients with temporal arteritis and polymyalgia rheumatica].
In 4 patients with temporal arteritis or polymyalgia rheumatica, women aged 60, 57, 83 and 73 years respectively, signs of aortic involvement were established. The first patient presented with signs of systemic inflammation without signs of temporal arteritis or aortitis. In the second, an acute symptomatic thoracoabdominal aneurysm developed. In the third, temporal arteritis was associated with chronic progressive dilatation ofthe thoracic aorta. The fourth developed signs of intermittent claudication of the extremities. The clinical manifestations in all patients were attributed to chronic inflammation of the aorta caused by giant cell arteritis. Aortic giant cell arteritis frequently accompanies temporal arteritis, but is rarely diagnosed. Up to 75% of patients with temporal arteritis may have some degree of aortic involvement. Thoracic aneurysms, complicated by rupture or dissection, are the most serious complications. Aortic disease associated with signs of systemic inflammation should trigger the suspicion of giant cell arteritis. Corticosteroids are the most important part of treatment. Three patients recovered following treatment; the first two received an endoprosthesis; in the woman aged 83 years, this was not technically possible; she died after 1.5 years.


Acute Kynurenine Exposure of Rat Thoracic Aorta Induces Vascular Dysfunction via Superoxide Anion Production.
The accumulation of uremic toxins is known to be one of the causes of cardiovascular disorder related to renal disease. Among the many uremic toxins, we focused on kynurenine (kyn), whose levels have been shown to be positively correlated with vascular endothelial dysfunction markers, and directly evaluated the influence of kyn on the rat thoracic aorta. Exposure of the endothelium-intact aorta to kyn markedly attenuated the acetylcholine (ACh)-induced relaxation and significantly increased superoxide anion (O2·-) production. These effects were ameliorated by pretreatment with ascorbic acid, an antioxidant, and CH223191, an aryl hydrocarbon receptor (AhR) inhibitor, but not by apocynin, a reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor. In the endothelium-denuded aorta, kyn significantly attenuated the nitric oxide (NO) donor sodium nitroprusside (SNP)-induced vasorelaxation and increased the O2·- production. Ascorbic acid treatment significantly ameliorated these effects, whereas CH223191 and apocynin treatments did not. Kyn had no influence on the vasorelaxant response to BAY 41-2272, a soluble guanylate cyclase stimulator. This suggested that kyn attenuates the NO-mediated vasorelaxation response by promoting O2·- production in thoracic aorta to inactivate NO. O2·- production is likely stimulated in both vascular endothelium and smooth muscle, the former of which may be mediated by AhR activation.


Surface photovoltage phase spectra for analysing the photogenerated charge transfer and photocatalytic activity of ZnFe2O4-TiO2 nanotube arrays.
The transfer properties of the photogenerated charge carriers in TiO2 nanotube arrays (TNAs) and ZnFe2O4 modified TiO2 nanotube arrays (ZFO-TNAs) were investigated by surface photovoltage phase spectra. The modification of ZnFe2O4 on TNAs will lead to a new process of photogeneration of charge carriers. When irradiated with UV or visible light, before ZnFe2O4 modification photogenerated holes can transfer to the surface of TNAs; while after ZnFe2O4 modification, a process of photogenerated electrons transferring to the surface occurred. The different detected photogenerated charge transfer processes agree well with the photocatalytic reduction of Cr(VI) and the degradation of methyl orange experiments, which confirmed that the photogenerated holes transferring to the surface of the TNAs or ZFO-TNAs were beneficial for the reactions of photocatalytic reduction and electrons transferring to the surface were beneficial for photocatalytic degradation.


Anti-neuroinflammatory capacity of fresh ginger is attributed mainly to 10-gingerol.
Despite the anti-neuroinflammatory capacity of ginger, the corresponding active constituents are unclear. This study analyzed the composition of fresh ginger ethanolic extract by using LC-MS. Inhibitory activities of fresh ginger extract and seven gingerol-related compounds on the neuro-inflammation were also evaluated by using a lipopolysaccharide (LPS)-activated BV2 microglia culture model. Except for zingerone and 6-gingerol, other gingerols and shogaols at a concentration of 20 μM inhibited the production of nitric oxide, IL-1β, IL-6 and TNF-α as well as their mRNA levels in LPS-activated BV2 microglia. Blocking NF-κB activation was the underlying mechanism responsible for inhibiting the proinflammatory gene expression. Increasing the alkyl chain length enhanced the anti-neuroinflammatory capacity of gingerols yet, conversely, attenuated those of shogaols. 6-Gingerol was the most abundant compound in the fresh ginger extract, followed by 10-gingerol. Furthermore, fresh ginger extract exhibited a significant anti-neuroinflammatory capacity, which was largely owing to 10-gingerol, but not 6-gingerol.


Grey zone score for hypo-osmotic swelling test (HOST) is not associated with embryo implantation defects.
To evaluate the clinical significance of the grey zone range (50-59%) of the hypo-osmotic swelling test (HOST) on sperm samples. Clinical and viable pregnancy rates and implantantion rates following in vitro fertilization-embryo transfer (IVF-ET) were determined in couples where the male partner scored between 50-59% on the HOST. The data were further evaluated according to the discrepancy between viability and HOST. Finally, the study assigned whether treating sperm with the protein digestive enzyme chymotrypsin had any effect on pregnancy outcome. Pregnancy and implantation rates were comparable for couples with grey zone HOST scores with the normal IVF rate for the center. Having a large discrepancy between viability and HOST scores had no clinical significance nor did treatment with chymotrypsin. In contrast to HOST scores < 50% where extremely poor implantation rates are noted, grey zone scores have no clinical significance.


Electrogenerated chemiluminescence derivatization reagents for carboxylic acids and amines in high-performance liquid chromatography using tris(2,2'-bipyridine)ruthenium(II).
2-(2-Aminoethyl)-1-methylpyrrolidine and N-(3-aminopropyl)pyrrolidine (NAPP) were found to be selective and sensitive derivatization reagents for carboxylic acid by high-performance liquid chromatography (HPLC) with electrogenerated chemiluminescence detection using tris(2,2'-bipyridine)ruthenium(II). Free fatty acids and ibuprofen were used as model compounds of carboxylic acids, and the derivatization conditions were optimized with myristic acid as a representative of free fatty acids. All the fatty acids tested were reacted with NAPP to produce highly sensitive derivatives under the mild reaction conditions of room temperature for 30 min in acetonitrile containing 2-bromo-1-ethylpyridinium tetrafluoroborate and 9-methyl-3,4-dihydro-2H-pyrido[1,2-a]pyrimidin-2-one. The chemiluminescence intensities were similar for all fatty acids. The derivatives obtained from 10 free fatty acids were completely separated by reversed-phase chromatography under isocratic elution conditions. The on-column detection limits (signal-to-noise ratio of 3) with proposed HPLC separation and chemiluminescence detection were 70 and 45 fmol for myristic acid and ibuprofen, respectively. The free fatty acids in human plasma were successfully determined using the present method. Histamine, a model compound of primary amines, was also determined after precolumn derivatization with 3-(diethylamino)propionic acid at room temperature for 60 min in acetonitrile containing N,N'-dicyclohexylcarbodiimide and 3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine with the detection limit of 70 fmol.


SVD and ASCII Character Encoding-Based Compression of Multiple Biosignals for Remote Healthcare Systems.
Advancements in electronics and miniaturized device fabrication technologies have enabled simultaneous acquisition of multiple biosignals (MBioSigs), but the area of compression of MBioSigs remains unexplored to date. This paper presents a robust singular value decomposition (SVD) and American standard code for information interchange (ASCII) character encoding-based algorithm for compression of MBioSigs for the first time to the best of our knowledge. At the preprocessing stage, MBioSigs are denoised, down sampled and then transformed to a two-dimensional (2-D) data array. SVD of the 2-D array is carried out and the dimensionality of the singular values is reduced. The resulting matrix is then compressed by a lossless ASCII character encoding-based technique. The proposed compression algorithm can be used in a variety of modes such as lossless, with or without using the down sampling operation. The compressed file is then uploaded to a hypertext preprocessor (PHP)-based website for remote monitoring application. Evaluation results show that the proposed algorithm provides a good compression performance; in particular, the mean opinion score of the reconstructed signal falls under the category "very good" as per the gold standard subjective measure.


Preoperative pulmonary risk assessment in esophagectomy.
Mortality rate after esophagectomy ranges from 2% to more than 25% and is related to preoperative and operative factors. Pulmonary complications are frequently seen after esophageal resection with a high mortality rate. The aim of preoperative pulmonary risk assessment is to identify patients who can undergo esophagectomy evaluating patient-related risk factors and their treatment. Preoperative pulmonary risk factors are smoking, respiratory diseases as COPD, elderly age and general condition. Careful medical history, physical examination and pulmonary function testing contribute to identify risk factors and strategies to reduce the risk of pulmonary complications.


Physical activity among minority populations: what health promotion practitioners should know--a commentary.
Leisure time and moderate-level physical activity participation in the United States is low among the majority of the population. Minority populations are especially inactive and report having fewer opportunities and access to be physically active. Physical activity programs utilizing a strong health promotion framework (including needs assessment, program plan and design, program implementation, and evaluation) are limited, particularly among minority populations. The purpose of this article is to discuss the importance of physical activity, briefly review and describe current literature, identify the need for physical activity programs among minority populations, and relate the concepts of a health promotion framework to physical activity programs that are usable among practitioners. The article also provides health promotion practitioners with various resources for improving physical activity programs.


A recombinant multi-epitope protein MEP1 elicits efficient long-term immune responses against HIV-1 infection.
The effective protective HIV vaccine should elicit either protective antibodies or effective T cell response, or both. To improve the efficacy of HIV-1 vaccines, HLA polymorphism and HIV-1 diversity are 2 key factors to be considered for vaccine development. In this study, we expressed a recombinant multi-epitope protein MEP1 which has the same amino acid sequence as a DNA vaccine for Chinese population in our previous report. We found that MEP1 alone could elicit moderate levels of humoral and cellular immune responses, but these responses could not provide protection from challenge with a recombinant virus rTTV-lucgag, which expresses Gag of HIV-1 CRF_07BC. Nevertheless, when MEP1 was immunized with aluminum adjuvant, both humoral and cellular immune responses were significantly increased, and they were protective against virus infection; meanwhile, MEP1 with aluminum not only elicited early (10 d post immunization) but also a long-term (at least 44 weeks post immunization) immune responses in BALB/c mice. These results suggested that MEP1 has the potential to be developed as an effective vaccine candidate, and that suitable adjuvant is necessary for this protein to generate protective immune responses.


A comparative study on the contraction induced by high K/Na deficient solution in the trachea, gall bladder and urinary bladder in guinea-pig.
In the present paper, a comparative study of high K solution was carried out on the mechanical response, wet and dry weights of tissue, and extracellular space of 14C-sorbitol in the trachea, gall bladder, and urinary bladder of guinea-pig. Hyperosmotic addition of KCl to the physiological solution produced a sustained contraction, while isosmotic substitution of K for Na in the physiological solution induced a contraction followed by a small sustained one in the trachea, gall bladder or urinary bladder. At 120 min after application of the substituted KCl, Na deficient solution, it increased cell water content in the smooth muscle cells of the trachea or gall bladder, but did not change that in the urinary bladder. Both the inhibition of contraction and the swelling in the trachea or gall bladder were prevented by the hyperosmotic application of sucrose or NaCl. The substitution for NaCl with the K salts with more permeable anions (I- greater than NO3- greater than Cl-) produced a greater inhibition of contraction in the trachea and gall bladder. Although the impermeable anion C2H5CO2- did not inhibit the contraction in the trachea, this anion partially inhibited it in the gall bladder. On the other hand, the inhibition of contraction in the urinary bladder was prevented by addition of pyruvate, oxalacetate or NaCl.(ABSTRACT TRUNCATED AT 250 WORDS)


Biofilm accumulation on additive manufactured Ti-6Al-4V alloy surfaces.
This study investigated whether additive manufactured (AM) surfaces inhibit accumulation of bacterial biofilm on the surfaces of Ti-6Al-4V alloy dental implants. Bacterial biofilms are thought to cause peri-implant disease, which develops in mucosa surrounding titanium (Ti) and Ti alloy dental implants and can lead to bone loss and implant failure. Accumulation of a Streptococcus mutans (ATCC 25175) biofilm on Ti-6Al-4V alloy was compared in relation to fabrication method, ie, AM using electron beam melting (EBM) or laser beam melting (LBM). Conventional lost-wax casting was used as positive control, and Teflon was used as negative control. Biofilm accumulation on the alloys and negative control (each n = 10) was conducted at 37°C under anaerobic conditions. After 4 h, the number of metabolically active S. mutans bacteria adhering to the alloy was determined with a bioluminescence assay. The quantitative roughness values of the specimens, before exposure to bacteria, ranked EBM > LBM > cast > Teflon. The amount of biofilm accumulation on the investigated AM metals and cast metal controls did not significantly differ.


The Wnt inhibitor dickkopf-1: a link between breast cancer and bone metastases.
Breast cancer is the second leading cause of cancer death in women and metastasizes to bone in greater than 80 % of advanced-disease patients. Once breast cancer bone metastases are established, the disease is incurable and drives numerous complications that increase morbidity and diminish patients' quality of life. Many mechanisms have been implicated in bone metastases of breast cancer. The critical role of Wnt signalling pathway inhibition in initiating bone lesions has been demonstrated in a variety of bone diseases and tumours. Overexpression of dickkopf-1 (Dkk1) protein, a negative regulator of the Wnt/β-catenin pathway, has been found in breast cancer cell lines that form osteolytic metastases preferentially and in serum from breast cancer patients with osteolytic bone metastases. Further understanding of the mechanistic role of Dkk1 as a link between primary breast tumours and secondary osteolytic bone metastases may facilitate development of anti-Dkk1 antibody therapeutic tools.


UGT1A1*28 polymorphism as a determinant of irinotecan disposition and toxicity.
The metabolism of irinotecan (CPT-11) involves sequential activation to SN-38 and detoxification to the pharmacologically inactive SN-38 glucuronide (SN-38G). We have previously demonstrated the role of UGT1A1 enzyme in the glucuronidation of SN-38 and a significant correlation between in vitro glucuronidation of SN-38 and UGT1A1 gene promoter polymorphism. This polymorphism (UGT1A1*28) is characterized by the presence of an additional TA repeat in the TATA sequence of the UGT1A1 promoter, ((TA)7TAA, instead of (TA)6TAA). Here we report the results from a prospective clinical pharmacogenetic study to determine the significance of UGT1A1*28 polymorphism on irinotecan disposition and toxicity in patients with cancer. Twenty patients with solid tumors were treated with a 90 min i.v. infusion of irinotecan (300 mg m(-2)) once every 3 weeks. The frequency of UGT1A1 genotypes was as follows: 6/6--45%, 6/7--35% and 7/7--20%, with allele frequencies of 0.375 and 0.625 for (TA)7TAA and (TA)6TAA, respectively. Patients with the (TA)7TAA polymorphism had significantly lower SN-38 glucuronidation rates than those with the normal allele (6/6>6/7>7/7, P = 0.001). More severe grades of diarrhea and neutropenia were observed only in patients heterozygous (grade 4 diarrhea, n = 1) or homozygous (grade 3 diarrhea/grade 4 neutropenia, n = 1 and grade 3 neutropenia, n = 1) for the (TA)7TAA sequence. The results suggest that screening for UGT1A1*28 polymorphism may identify patients with lower SN-38 glucuronidation rates and greater susceptibility to irinotecan induced gastrointestinal and bone marrow toxicity.


Limited role of biochars in nitrogen fixation through nitrate adsorption.
Nitrate cycling is essential in sustaining soil systems. Excessive application of N-fertilizers and the associated underground water contamination have attracted a great deal of research attention. Sorption is efficient and environmentally friendly in nitrate fixation. A debate was noted in literature regarding whether biochars have potential to fix nitrate through sorption. In this study, biochars produced from different biomasses as well as biomass compositions were chosen as the absorbents to evaluate their potential efficiencies in nitrate fixation. Increased sorption to nitrate was observed for biochars with increased pyrolysis temperature, but the increasing extent varied with biomass. The surface base functional groups and surface charges of biochars could not well explain nitrate sorption. The significant positive correlation between nitrate sorption and biochar surface areas suggested that surface area was the controlling parameter for nitrate sorption. The pre-coating of tannic acid (TA) on biochars decreased but did not completely inhibit nitrate sorption. This observation suggested that nitrate sorption on biochars may be further decreased after their interactions with natural organic matter. Nitrate sorption was compared among various adsorbents, including biochars, soil particles, clay minerals, engineered particles, as well as humic substances. Soil particles generally showed high sorption to nitrate over biochars. This result suggested that biochars investigated in this work may play a limited role in nitrate fixation through sorption after their massive application. Nitrogen fixation through nitrate adsorption on biochars should be carefully evaluated taking into consideration of biochar feedstocks and properties.


Assessment of doses to the offspring of the Techa River cohort due to intakes of radionuclides by the mother.
The Techa River was contaminated as a result of radioactive releases by the Mayak plutonium production facility in 1949-1956. The residents of riverside communities were exposed to internal irradiation from radionuclides ingested mainly with river water, and also to external gamma irradiation resulting from shoreline and flood-plain contamination. The most important role in population exposure was played by (89,90)Sr and 137Cs. The persons born after the onset of the contamination have been identified as the 'Techa River Offspring Cohort' (TROC). The TROC has the potential to provide direct data on health effects in progeny that resulted from exposure of a general population to chronic radiation. This report describes the results of the calculation of fetal doses due to intakes of radionuclides by their mothers. Particular attention has been given to fetal dose from 90Sr because this nuclide is the most significant in terms of population dose for the Techa River. The comparison of the fetal bone marrow doses evaluated using different approaches proposed in the literature has shown a large dispersal in dose values. The main cause of this is the difference in model assumptions simplifying some developmental aspects of fetal haematopoiesis and bone formation. This paper presents an analysis of these basic assumptions that could be useful for further improvements in fetal dosimetry.


Adrenal paraneurone contractile proteins and stimulus-secretion coupling.
Actin, myosin, and alpha-actinin have been isolated from adrenal chromaffin cells and characterized. Their physicochemical properties have been studied and their cell localization revealed by biochemical, immunocytochemical, and ultrastructural techniques. Alpha-actinin is a component of chromaffin granule membranes and some of the cell actin copurifies with these secretory granules. Myosin is not detected in the granules but is present mainly in the cytosol. Trifluoperazine, a calmodulin antagonist, blocks stimulation-induced hormone release from chromaffin cells at a step distal from Ca2+ entry. High affinity calmodulin binding sites have also been found in chromaffin granule membranes. Furthermore, microinjection of calmodulin antibodies into chromaffin cells blocks hormone output in response to stimulation. In view of the above findings, the possible roles of contractile proteins and calmodulin in chromaffin cell functions is discussed.


Prevalence of Parkinson's disease in Korea.
The prevalence of neurodegenerative disorders is not well documented in Korea. We assessed the prevalence of Parkinson's disease in an elderly population in a newly industrialized city in a rural region. Subjects for this study were randomly selected from a community-based cohort study. The sample in the cohort represented approximately 1.3% (4700) of 362 625 adults (age>18 years) listed in the city register in 1998. Among this group, 4218 subjects (1086 subjects aged>60 years) agreed to be interviewed and underwent a physical examination and neuropsychological tests administered by a neurologist and neuropsychologist. All participants were examined. Participants who had bradykinesia and at least one other possible cardinal sign of parkinsonism at the neurologic screening, and those who reported that they had Parkinson's disease, or were taking antiparkinsonian drugs were identified. In our study, 16 subjects showed evidence of Parkinson's disease. The prevalence in this population was 0.37%. Prevalence increased with age, and prevalence was 1.47% for those aged older than 60 years. Postural instability and gait disturbance were more common in the older age group. The results of neuropsychological tests were as follows: (1) only two subjects had low scores (<20) in the Korea-version mini-mental status examination; (2) seven subjects scored 0.5, one subject scored 2 and the other eight subjects scored 0 in the clinical dementia rating. The results of our prevalence study are similar to those of studies carried out in Western countries. Age is a risk factor for Parkinson's disease in Korea.


The Year in Extracorporeal Membrane Oxygenation: Selected Highlights From 2021.
This review summarizes the extracorporeal membrane oxygenation (ECMO) or extracorporeal life support literature published in 2021. This Selected Highlights article is not intended to be an exhaustive review of the literature, but rather a summarizing of key themes that developed in the ECMO literature during 2021. The primary topics presented include the following: ECMO for coronavirus disease 2019, extracorporeal cardiopulmonary resuscitation, periprocedural cardiopulmonary support with ECMO, and anticoagulation for ECMO.


The impact of Tegillarca granosa extract haishengsu on HL-60 cell.
Haishengsu (Hss) is a purified protein from Tegillarca granosa that has been used as a traditional Chinese medicine to treat cancer for more than a century. In this study, we observed the impact of Haishengsu (Hss) on the proliferation and differentiation of HL-60 cells in the leukemic cell line by taking tretinoin and AS2O3 as a positive control and making a comparative analysis between the effect of Hss and tretinoin and AS2O3. We found that Hss could significantly inhibit the proliferation of HL-60 cells and caused most of the cells to stay in the G0/G1 phase. Its effect was much stronger than that of tretinoin and AS2O3, and the ability of Hss to induce differentiation was close to tretinoin. Hss functions probably by inhibiting the expression of the Bcl-2 and MPO genes and further promoting the expression of the Bax gene. Hss has a significant effect on both inhibiting the proliferation and inducing the differentiation of HL-60 cells. It is possible that Hss may be a new kind of clinical differentiation inducer.


Elevation of the adenylate pool in rat cardiomyocytes by S-adenosyl-L-methionine.
Rapid resynthesis of the adenylate pool in cardiac myocytes is important for recovery of contractility and normal function of regulatory mechanisms in the heart. Adenosine and adenine are thought to be the most effective substrates for nucleotide synthesis, but the possibility of using other compounds has been studied very little in cardiomyocytes. In the present study, the effect of S-adenosyl-L-methionine (SAM) on the adenylate pool of isolated cardiomyocytes was investigated and compared to the effect of adenine and adenosine. Adult rat cardiomyocytes were isolated using the collagenase perfusion technique. The cells were incubated in the presence of adenine derivatives for 90 min followed by nucleotide determination by HPLC. The concentrations of adenine nucleotides expressed in nmol/mg of cell protein were initially 22.1 +/- 1.4, 4.0 +/- 0.3 and 0.70 +/- 0.08 for ATP, ADP and AMP, respectively (n = 10, +/- S.E.M.), and the total adenylate pool was 26.8 +/- 1.6. In the presence of 1.25 mM SAM in the medium, the adenylate pool increased by 5.2 +/- 0.4 nmol/mg of cell protein, but only if 1 mM ribose was additionally present in the medium. No changes were observed with SAM alone. A similar increase (by 4.9 +/- 0.6 nmol/mg protein) was observed after incubation with 1.25 mM adenine plus 1 mM ribose, but no increase was observed if ribose was omitted. Adenosine at 0.1 or 1.25 mM concentrations also caused an increase in the adenylate pool (by 5.2 +/- 1.0 and 5.2 +/- 0.9 nmol/mg protein, respectively), which in contrast to the SAM or adenine was independent of the additional presence of ribose. Thus, S-adenosyl-L-methionine could be used as a precursor of the adenylate pool in cardiomyocytes, which is as efficient in increasing the adenylate pool after 90 min of incubation as adenosine or adenine. Nucleotide synthesis from SAM involves the formation of adenine as an intermediate with its subsequent incorporation by adenine phosphoribosyltransferase.


Exploring the mesenchymal stem cell niche using high throughput screening.
In the field of stem cell technology, future advancements rely on the effective isolation, scale-up and maintenance of specific stem cell populations and robust procedures for their directed differentiation. The stem cell microenvironment - or niche - encompasses signal inputs from stem cells, supporting cells and from the extracellular matrix. In this context, the contribution of physicochemical surface variables is being increasingly recognised. This paradigm can be exploited to exert control over cellular behaviour. However, the number of parameters at play, and their complex interactions, presents a formidable challenge in delineating how the decisions of cell fate are orchestrated within the niche. Additionally, in the case of mesenchymal stem cells (MSC), more than one type of stem cell niche has been identified. By employing high throughput screening (HTS) strategies, common and specific attributes of each MSC niche can be probed. Here, we explore biological, chemical and physical parameters that are known to influence MSC self-renewal and differentiation. We then review techniques and strategies that allow the HTS of surface properties for conditions that direct stem cell fate, using MSC as a case study. Finally, challenges in recapturing the niche, particularly its three dimensional nature, in surface-based HTS formats are discussed.


Augmentation with an ovine forestomach matrix scaffold improves histological outcomes of rotator cuff repair in a rat model.
Rotator cuff tears can cause significant pain and functional impairment. Without surgical repair, the rotator cuff has little healing potential, and following surgical repair, they are highly prone to re-rupture. Augmenting such repairs with a biomaterial scaffold has been suggested as a potential solution. Extracellular matrix (ECM)-based scaffolds are the most commonly used rotator cuff augments, although to date, reports on their success are variable. Here, we utilize pre-clinical in vitro and in vivo assays to assess the efficacy of a novel biomaterial scaffold, ovine forestomach extracellular matrix (OFM), in augmenting rotator cuff repair. OFM was assessed in vitro for primary tenocyte growth and adherence, and for immunogenicity using an assay of primary human dendritic cell activation. In vivo, using a murine model, supraspinatus tendon repairs were carried out in 34 animals. Augmentation with OFM was compared to sham surgery and unaugmented control. At 6- and 12-week time points, the repairs were analysed biomechanically for strength of repair and histologically for quality of healing. OFM supported tenocyte growth in vitro and did not cause an immunogenic response. Augmentation with OFM improved the quality of healing of the repaired tendon, with no evidence of excessive inflammatory response. However, there was no biomechanical advantage of augmentation. The ideal rotator cuff tendon augment has not yet been identified or clinically implemented. ECM scaffolds offer a promising solution to a difficult clinical problem. Here, we have shown improved histological healing with OFM augmentation. Identifying materials that offset the poorer mechanical properties of the rotator cuff post-injury/repair and enhance organised tendon healing will be paramount to incorporating augmentation into surgical treatment of the rotator cuff.


Seminal reservoirs during an HIV type 1 eradication trial.
Despite dramatic reduction of the levels of human immunodeficiency virus type I (HIV-1) virions in blood and seminal plasma of infected patients, highly active antiretroviral therapy (HAART) does not eradicate HIV-1. Three patients, with less than 50 copies/ml of plasma viral RNA, were enrolled in this eradication protocol. Didanosine (DDI) and hydroxyurea (HU) were added to their baseline HAART and after a month of therapy, low dose OKT3, followed by a 2-week course of interleukin 2 (IL-2), was administrated. All antiretroviral therapy was then interrupted and the three patients developed viral rebound in the peripheral blood. The V3 loop region of the HIV-1 gp120 from cell-free viral RNA and proviral DNA in blood and seminal compartments was sequenced in one patient. The two major viral isolates in semen cells were macrophage- tropic (R5) and dual-tropic (R5X4), and these isolates were also present in the PBMCs. Six months after the viral rebound, we demonstrated a shift toward dual tropism in semen cell-associated HIV-1 proviral DNA, with the first appearance of a T-lymphotropic (X4) provirus solely in this compartment. The virus responsible for the blood plasma viral rebound was never found in the semen microenvironment. This study suggests viral compartmentalization of the semen microenvironment after an intensification and stimulatory HIV-1 eradication protocol, with evidence of viral evolution.


Evaluation and surgical management of intrinsic sphincter deficiency after radical prostatectomy.
Urinary incontinence following prostatectomy is usually due to intrinsic sphincter deficiency and is often referred to as post-prostatectomy incontinence (PPI). The male sling is an effective minimally invasive procedure for low volume PPI. Although the male sling procedure is becoming increasingly popular, the artificial urinary sphincter (AUS) remains the gold standard. Placement of the AUS cuff using the transcorporal technique salvages patients with urethral atrophy as well as prior AUS erosion or infection. As the surgical options for PPI expand, it is important to analyze the outcomes with contemporary surgical techniques and to develop an algorithm for procedure selection.


Recent myocardial infarction: assessment with unenhanced T1-weighted MR imaging.
The purpose of the study was to prospectively evaluate a T1-weighted technique for detection of myocardial edema resulting from recent myocardial infarction (MI) or intervention. This study was HIPAA compliant and institutional review board approved. Fifteen men and one woman (mean age, 57.8 years+/-11.5 [standard deviation]) were examined with T1-weighted magnetic resonance (MR) imaging and inversion-recovery cine pulse sequence in two groups, recent MI and chronic MI, and gave informed consent. T1 relaxation times of MI and adjacent myocardium were compared (Student t test and correlation analysis). In patients with recent MI, areas of myocardial edema were well depicted with T1-weighted MR imaging. T1 relaxation times of recent infarcts were longer than those of older MIs (925 msec+/-169 vs 551 msec+/-107, P<.001). From local edema, T1 relaxation time of infarcted myocardium is increased, may remain elevated for 2 months, and enables imaging with T1-weighted techniques.


[Genetic analysis of hantaviruses carried by Rattus norvegicus collected from Henan and Neimenggu provinces, China].
Genetic analysis was performed to infer the relationship between hantaviruses carried by Rattus norvegicus from Henan and Neimenggu provinces and the other known hantavirus and the vaccine strain. Total RNA was extracted from lung tissues with Trizol reagent. The complete M and S segment sequences of strains NM133 and Q12 were amplified by RT-PCR. The purified DNA fragments were directly subjected to sequencing, and then to sequence analysis and phylogenetic analysis. The complete S segment sequences of strains NM133 and Q12 were found to be 1770 nt and 1772 nt in length respectively, with one open reading frame encoding 429 amino acids. The complete M segment sequences of both two strains are 3654 nucleotide in length encoding a protein of 1133 amino acids. The two strains shared a high degree of homology with most of known Seoul virus (SEOV) but quite different from Hantaan virus and other hantaviruses. Furthermore, the nucleoprotein and glycoprotein of the two strains had the congruent structure with the vaccine strain Z37. On the S- and M-phylogenetic trees, both strains (NM133 and Q12) were grouped into the first cluster of SEOV, and were more closely related to the strains, such as: Hb8610, R22, HB55, L99, and K24-e7. Both strains (NM133 and Q12) belonged to SEOV, and sharing a high degree of homology and similar secondary structure with strains including the vaccine strains Z37, our data suggested that the present vaccine used in China could effectively prevent HFRS caused by SEOV.


Cerebellar vermis H₂ receptors mediate fear memory consolidation in mice.
Histaminergic fibers are present in the molecular and granular layers of the cerebellum and have a high density in the vermis and flocullus. Evidence supports that the cerebellar histaminergic system is involved in memory consolidation. Our recent study showed that histamine injections facilitate the retention of an inhibitory avoidance task, which was abolished by pretreatment with an H2 receptor antagonist. In the present study, we investigated the effects of intracerebellar post training injections of H1 and H2 receptor antagonists as well as the selective H2 receptor agonist on fear memory consolidation. The cerebellar vermi of male mice were implanted with guide cannulae, and after three days of recovery, the inhibitory avoidance test was performed. Immediately after a training session, animals received a microinjection of the following histaminergic drugs: experiment 1, saline or chlorpheniramine (0.016, 0.052 or 0.16 nmol); experiment 2, saline or ranitidine (0.57, 2.85 or 5.07 nmol); and experiment 3, saline or dimaprit (1, 2 or 4 nmol). Twenty-four hours later, a retention test was performed. The data were analyzed using one-way analysis of variance (ANOVA) and Duncan's tests. Animals microinjected with chlorpheniramine did not show any behavioral effects at the doses that we used. Intra-cerebellar injection of the H2 receptor antagonist ranitidine inhibited, while the selective H2 receptor agonist dimaprit facilitated, memory consolidation, suggesting that H2 receptors mediate memory consolidation in the inhibitory avoidance task in mice.


Pharmacokinetics of methotrexate in adult Indian patients and its relationship to nutritional status.
Since anticancer drugs have a low therapeutic index, any significant change in the pharmacokinetics of a drug would have a bearing on therapeutic efficacy and drug toxicity. Nutritional deficiencies have been shown to affect the four processes of pharmacokinetics, ie, absorption, distribution, biotransformation, and excretion. Malnutrition and undernutrition are widely prevalent in India and thus it was thought to be of practical importance to study the effect of initial nutritional status on the overall kinetics of methotrexate, a widely used anticancer drug. The results of the study reveal that relative weight correlates well with the anthropometric parameters, nutritional parameters, and dietary intake and may be used as a marker of nutritional status. When grouped on the basis of relative weights, the undernourished patients revealed a significant (P less than 0.01) prolongation of biological half-life and a significant (P less than 0.001) reduction in clearance. Based on these results, relative weight has been proposed as a basis for drug dosage determinations in place of the existing practice of administering antineoplastic drugs on the basis of body surface area or body weight alone.


Formulary decisions for pre-1938 medications.
A process is described for formulary revisions consistent with the Food and Drug Administration (FDA)'s current initiative to ensure that all drugs marketed in the United States have been approved for safety and efficacy. A list of pre-1938 drugs (i.e., formulations marketed before the Federal Food, Drug, and Cosmetic Act established safety requirements) was compiled from USP DI Volume III and International Journal of Pharmaceutical Compounding lists. Products on the resulting list were reviewed for current marketing and FDA approval status and for availability. The project team recommended formulary addition or retention if a product had been used and been purchased more than once at the hospital in the past year, if no formulary alternative was available, or if the product had been approved by FDA. Nonformulary status was recommended if none of these criteria applied or the product was no longer available. Of 88 pre-1938 formulations of 59 drugs, only 3 had been approved by FDA before 1962 (when evidence of efficacy was first required) and 14 thereafter. Of the 88 formulations, 47 were on the hospital's formulary. The team recommended that 37 formulations be retained on or added to the formulary and that 51 be maintained or designated as nonformulary. The hospital's pharmacy and therapeutics (P&T) committee accepted the recommendations, provided that the 30 nonapproved formulations recommended for formulary status be reviewed as FDA continues its effort to have manufacturers either apply for approval of their products or remove them from the market. The recommendations were also accepted by the health system's P&T committee. A systematic approach to reviewing pre-1938 medications for the purpose of formulary revision was successful in addressing safety concerns about these older drug formulations.


Pseudoexfoliation syndrome without glaucoma: White matter abnormalities detected by conventional MRI and diffusion tensor imaging.
To assess macro- and microstructural brain changes in patients with pseudoexfoliation syndrome (PXS). Comprehensive ophthalmic examination and brain MRI were conducted on 20 patients with PXS without glaucoma (aged 62.75 ± 0.4 years) and 20 controls (aged 62 ± 0.6 years). White matter (WM) integrity was evaluated on FLAIR and single-shot multisection SE-EPI diffusion tensor imaging (DTI) sequences. The presence and the number of white matter hyperintensities (WMHIs) on FLAIR images was compared between all patients and control subjects. Microstructural WM changes on DTI was evaluated using Tract-based spatial statistics (TBSS). DTI metrics of the optic tracts were assessed by the region-of-interest (ROI) method. A significantly higher number of WMHIs was found in the patients with PXS than in the control subjects (P ≤ 0.002). On DTI the patients showed bilateral increase in the mean diffusivity (MD), axial diffusivity (AD) and radial diffusivity (RD) values in the anterior thalamic radiation, the inferior fronto-occipital fasciculus, the superior longitudinal fasciculus, the inferior longitudinal fasciculus and the forceps minor. TBSS revealed no significant difference in fractional anisotropy (FA) values, but ROIs analysis of the optic tracts revealed decreased FA values in the patients. MRI in patients with PXS detects abnormalities in the brain and the optic tracts at a subclinical stage. Early detection of microstructural changes could be useful to guide appropriate treatment to impede the disease process.


Cruciferous Vegetables as Antioxidative, Chemopreventive and Antineoplasic Functional Foods: Preclinical and Clinical Evidences of Sulforaphane Against Prostate Cancers.
Sulforaphane (SF, 1-isothiocyanato-4-(methyl-sulfinyl)-butane) is found in broccoli, cabbage and cauliflower. we performed a critical review on the antioxidative, chemopreventive and antitumor effects of SF from cruciferous vegetables against prostate cancers and molecular pathways. For a complete and reliable review, primary and secondary resources were used, including original and review articles, books and government documents published until March 2018. Articles that are in duplicity and disconnected are not considered for review. SF is derived from glucoraphanin (4-methyl-sulfinyl-butyl-glucosinate), being one of the most commonly found isothiocyanates in vegetables from Brassica spp., especially in broccoli samples. In vitro studies indicate that SF induces apoptosis in a dependent or non-dependent method of androgens by transcription of tumor suppressor genes, oxidation response and higher expression of phase II enzymes in prostate cancer cells. Sulforaphane also decreases transcription of the nuclear factor kB and antiapoptotic proteins, expression of cyclin D2 and survivin and DNA synthesis, increases Nrf2 gene activity, interferes with genome compacting by inhibition of histone deacetylases and disrupts Hsp90 complexes, which cause cell cycle arrest, mitosis interruption, activation of caspases and mitochondria depolarization. SF and cruciferous vegetables play antioxidative and chemopreventive role, delaying or blocking in vivo carcinogenesis, causing biochemical and epigenetic changes, preventing, delaying, or reversing preneoplastic or advanced prostate lesions, and frequently activating tumor cell death by intrinsic methods of apoptosis. These outcomes encourage the consumption of Brassica specimens, which could be easily achieved by the incorporation of food and vegetables rich in cruciferous isothiocyanates in the diet.


An in vitro bioassay for nerve growth factor based on 24-hour survival of PC-12 cells.
We have developed an in vitro bioassay for nerve growth factor (NGF) which offers several advantages over currently used alternatives. The assay uses the commercially available PC-12 cell line, and requires only 24 hr incubation with NGF. The cells do not require priming, and are dosed as they are originally seeded in serum-free medium in 96-well cell culture plates coated with collagen. Concentrations between one and 20 ng/ml NGF can be measured, and only standard cell culture equipment and a microplate reader are required.


Muscarinic M2 receptor is active on pancreatic islets from hypothalamic obese rat.
Hypothalamic obese rats, obtained by neonatal treatment with monosodium L-glutamate (MSG), are hyperinsulinemic, and secrete more insulin than lean ones do when stimulated by glucose, while acetylcholine insulinotropic effect decreases. The effect of acetylcholine on glucose-induced insulin secretion is attributed to muscarinic receptors of pancreatic beta cells, mainly to M(3) subtype. However, it has been observed that activation of M(2) or M(4) subtypes causes inhibition of glucose-induced insulin secretion in insulin secreting cell line. Insulin secretion was measured, stimulated by glucose in the presence of acetylcholine plus methoctramine, a muscarinic M(2) antagonist, on pancreatic islets isolated from MSG-obese and lean rats to investigate whether impairment of acetylcholine insulinotropic effect on pancreatic islets from MSG-obese rats has any relationship with muscarinic M(2) receptor function in beta cells. Insulin secretion stimulated by 8.3 mM glucose was higher in islets from obese rats than from lean ones. Insulinotropic effect of acetylcholine was reported in islets of both animals, albeit less than in obese ones. Blockage of muscarinic M(2) receptor, using methoctramine at 1; 5 and 10 microM, increased acetylcholine secretory response in islets of obese rats, while no effect has been observed in lean ones. Results demonstrate that muscarinic M(2) receptors are functioning in pancreatic islets of MSG-obese rats. The inhibitory action of muscarinic M(2) receptor may be a mechanism by which acetylcholine discloses weak insulinotropic effect in MSG-obese rats.


In vivo effects of DL-alpha-difluoromethylornithine on the polyamine and nucleotide phosphate metabolism in P388/S leukemia cells.
In vivo effects of DL-alpha-difluoromethylornithine (DFMO) on the metabolism of polyamines and nucleotide phosphates were monitored in P388/S leukemia cells grown intraperitoneally in BDF1 inbred male mice. Inhibiting the ornithine decarboxylase (ODC) activity DFMO depleted putrescine and spermidine to 30-50 and 50-60%, respectively, and increased spermine to 25-60% compared with the controls, when given as 2% solution in drinking water of the tumor-bearing animals. DFMO treatment caused a parallel 56% elevation of total nucleotide content in tumor cells with distinct and significant increase of some nucleotide phosphates. The most pronounced alterations were shown in the intracellular UTP (202%), CTP (103%), ADP (92%) and ATP (71%) concentrations. Changes in polyamine and nucleotide phosphate metabolisms were dependent on tumor progression. A possible explanation of the metabolic events induced by DFMO is discussed.


Li Ion Conducting Polymer Gel Electrolytes Based on Ionic Liquid/PVDF-HFP Blends.
Ionic liquids thermodynamically compatible with Li metal are very promising for applications to rechargeable lithium batteries. 1-methyl-3-propylpyrrolidinium bis(trifluoromethanesulfonyl)imide (P(13)TFSI) is screened out as a particularly promising ionic liquid in this study. Dimensionally stable, elastic, flexible, nonvolatile polymer gel electrolytes (PGEs) with high electrochemical stabilities, high ionic conductivities and other desirable properties have been synthesized by dissolving Li imide salt (LiTFSI) in P(13)TFSI ionic liquid and then mixing the electrolyte solution with poly(vinylidene-co-hexafluoropropylene) (PVDF-HFP) copolymer. Adding small amounts of ethylene carbonate to the polymer gel electrolytes dramatically improves the ionic conductivity, net Li ion transport concentration, and Li ion transport kinetics of these electrolytes. They are thus favorable and offer good prospects in the application to rechargeable Li batteries including open systems like Li/air batteries, as well as more "conventional" rechargeable lithium and lithium ion batteries.


Production and recognition bias of stylistic sentences using a story reading task.
Four experiments examined participants' ability to produce surface characteristics of sentences using an on-line story reading task. Participants read a series of stories in which either all, or the majority of sentences were written in the same "style," or surface form. Twice per story, participants were asked to fill in a blank consistent with the story. For sentences that contained three stylistic regularities, participants imitated either all three characteristics (Experiment 2) or two of the three characteristics (Experiment 1), depending on the proportion of in-style sentences. Participants demonstrated a recognition bias for the read style in an unannounced recognition task. When participants read stories in which the two styles were the dative/double object alternation, participants demonstrated a syntactic priming effect in the cloze task, but no consistent recognition bias in a later recognition test (Experiments 3 and 4).


Relationship between family caregiver quality of life and the care provided to people living with dementia: protocol for a mixed methods study.
Family caregivers of people with dementia perform duties that are important for maintaining their loved one's overall well-being. However, it is not yet clear how these caregivers' quality-of-life affects their ability to meet the care demands of their loved ones. The purpose of this study is to utilize a mixed methods approach in investigating how family caregiver quality-of-life affects the care provided to people with dementia. Family caregivers will be recruited from the Rochester, New York area to participate in focus groups or one-on-one interviews. In addition to the qualitative data obtained, caregivers will complete questionnaires regarding their own quality-of-life (e.g., health status, socioeconomic) as well as the care provided to their loved ones with dementia (e.g., how the care provided meets the needs of the care recipient, time spent). A convergent mixed methods approach will be used to analyze the qualitative and quantitative data obtained. Data from the interviews will be transcribed verbatim and then analyzed qualitatively. Quantitative data from the questionnaires will be analyzed using IBM SPSS Statistics software. A convergent mixed methods approach will be applied to the datasets to help shed light on the relationship between family caregiver quality-of-life and the care provided to people living with dementia. Understanding of this relationship will make it possible to develop initiatives that better address caregiver needs.


Seat inclination, use of lumbar support and low-back pain of taxi drivers.
Epidemiologic evidence supporting optimal seating is limited and inconsistent. This cross-sectional study was conducted to examine the association between seat inclination, use of lumbar support, and the prevalence of clinically significant low-back pain among taxi drivers A digital inclinometer was used to measure inclinations of seat surfaces (theta(seat)) and backrests (theta(back)), and calculate the back-to-thigh angle (theta(back-thigh)). Structured interviews were conducted to gather information on the use of lumbar support and the prevalence of low-back pain that had led to medical attention or absence from driving in the past month. A multiple logistic regression analysis was used to estimate the prevalence odds ratio (OR) with adjustment for age, body mass index, professional seniority, monthly driving hours, and the intensity of exposure to whole-body vibration. Among 224 drivers, the mean theta(seat), and theta(backrest) were 14.5 (SD 9.6) and 95.1 (SD 2.7) degrees, respectively, resulting in theta(back-thigh) of 80.6 (SD 9.3) degrees. Fifty-five percent used a lumbar support regularly, but 25% reportedly had significant low-back pain. The prevalence of low-back pain was 23% among those with theta(back-thigh) <86 degrees, 37% for those with a theta(back-thigh) of 86 approximately 91 degrees, and 9% for a theta(back-thigh) of >91 degrees. The adjusted OR comparing those with a theta(back-thigh) of < or = 91 degrees to those with a theta(back-thigh) of >91 degrees was 5.11 [95% confidence interval (95% CI) 1.07 approximately 24.4]. For regularly using drivers versus those not using lumbar support, the prevalence of low-back pain was 18% versus 34%, with an adjusted OR of 0.33 (95% CI 0.16 approximately 0.68). Neither theta(seat) nor theta(backrest) alone was significantly associated with low-back pain. The epidemiologic observation of this study was consistent with the results of prior biomechanical studies on appropriate seat inclinations and the use of lumbar support. Prospective studies are needed to confirm the true beneficial effects of these seating parameters.


Long-term complete remission in a patient with intravascular large B-cell lymphoma with central nervous system involvement.
This report describes a patient with intravascular large B-cell lymphoma (IVLBCL) with central nervous system involvement at the time of diagnosis who achieved complete remission for over 5 years in response to therapy. The patient, a 71 year-old woman, was previously healthy with the exception of taking verapamil for paroxysmal supraventricular tachycardia. She had presented with pyrexia and gradually progressive anemia. Brain magnetic resonance imaging revealed an infarct-like lesion in the pons, although no paralysis was observed. She was diagnosed with IVLBCL on the basis of random skin biopsy. After eight cycles of rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone therapy, abnormal laboratory data had normalized, and no pontine lesion was evident on magnetic resonance imaging without receiving any intrathecal chemotherapy. IVLBCL is associated with poor prognosis, particularly in patients with central nervous system involvement. Early initiation of rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone therapy and drug interactions between anticancer agents and verapamil as a p-glycoprotein inhibitor were considered the possible reasons for favorable outcome in the present case.


Alcohol Dependence Disrupts Amygdalar L-Type Voltage-Gated Calcium Channel Mechanisms.
L-type voltage-gated calcium channels (LTCCs) are implicated in several psychiatric disorders that are comorbid with alcoholism and involve amygdala dysfunction. Within the amygdala, the central nucleus (CeA) is critical in acute alcohol's reinforcing actions, and its dysregulation in human alcoholics drives their negative emotional state and motivation to drink. Here we investigated the specific role of CeA LTCCs in the effects of acute alcohol at the molecular, cellular physiology, and behavioral levels, and their potential neuroadaptation in alcohol-dependent rats. Alcohol increases CeA activity (neuronal firing rates and GABA release) in naive rats by engaging LTCCs, and intra-CeA LTCC blockade reduces alcohol intake in nondependent rats. Alcohol dependence reduces CeA LTCC membrane abundance and disrupts this LTCC-based mechanism; instead, corticotropin-releasing factor type 1 receptors (CRF1s) mediate alcohol's effects on CeA activity and drive the escalated alcohol intake of alcohol-dependent rats. Collectively, our data indicate that alcohol dependence functionally alters the molecular mechanisms underlying the CeA's response to alcohol (from LTCC- to CRF1-driven). This mechanistic switch contributes to and reflects the prominent role of the CeA in the negative emotional state that drives excessive drinking.SIGNIFICANCE STATEMENT The central amygdala (CeA) plays a critical role in the development of alcohol dependence. As a result, much preclinical alcohol research aims to identify relevant CeA neuroadaptions that promote the transition to dependence. Here we report that acute alcohol increases CeA neuronal activity in naive rats by engaging L-type calcium channels (LTCCs) and that intra-CeA LTCC blockade reduces alcohol intake in nondependent rats. Alcohol dependence disrupts this LTCC-based mechanism; instead, corticotropin-releasing factor type 1 receptors (CRF1s) mediate alcohol's effects on CeA activity and drive the escalated alcohol intake of alcohol-dependent rats. This switch reflects the important role of the CeA in the pathophysiology of alcohol dependence and represents a new potential avenue for therapeutic intervention during the transition period.


Subcorneal pustular dermatosis associated with Coccidioides immitis.
Coccidioidomycosis (AKA "Valley fever") is a primary pulmonary infection via airborne spores released from coccidioides immitis in the soil. Reactive cutaneous eruptions resulting from the pulmonary infection are difficult to diagnose because skin biopsies do not contain the organism. We present an adolescent male with primary pulmonary C.immitis infection manifesting with biopsy proven subcorneal pustular dermatosis. Serological studies revealed increasingly positive titers for coccidioidomycosis and symptoms resolved promptly following initiation of systemic antifungal therapy. Our unique case presentation illustrates subcorneal pustular dermatosis as a reactive eruption owing to primary pulmonary coccidioidomycosis. An association between the two conditions warrants further investigation.


Potential of Autologous Progenitor Cells and Decellularized Porcine Artery Matrix in Construction of Tissue-engineered Vascular Grafts.
To develop a tissue-engineered vascular graft, we used pericardial effusion-derived progenitor cells (PEPCs) collected from drained fluid after open-heart surgery in children with congenital heart diseases to repopulate a decellularized porcine pulmonary artery. The PEPCs were compared with human fibroblasts (HS68) and human umbilical vein endothelial cells (HUVECs) in cell growth and migration. They were cultured with the matrices via an inner approach (intima), lateral approach (media), and outer approach (adventitia). PEPCs grew and migrated better than the other two cells 14 days after seeding in the decellularized vessel. In immunofluorescence assays, PEPCs expressed CD90 and CD105 indicating a vascular differentiation. PEPCs grew in a decellularized porcine pulmonary artery matrix may have the potential for producing tissue-engineered vascular grafts.


Anophthalmia, Global Developmental Delay, and Severe Dysphagia in a Young Girl With 14q22q23 Microdeletion Syndrome.
14q22q23 microdeletion syndrome, also called Frias syndrome, is an extremely rare partial deletion of the long arm of chromosome 14 characterized by the anomalies of the pituitary gland, eyes, and hand/foot. Intellectual disability and facial dysmorphism are other common manifestations. Haploinsufficiency of the genes bone morphogenetic protein 4 (BMP4) and orthodenticle homeobox 2 (OTX2) accounts for most of the phenotypic abnormalities seen in these patients. There are only a few cases reported with Frias syndrome in the literature, and there are multiple variations present, which are not well recognized due to different set of genes involved. This case report presents the case of a young child with a deletion in 14q22.2q23.1 region containing both BMP4 and OTX2 genes as well as sineoculis homeobox homolog 1 (SIX1) and sineoculis homeobox homolog 6 (SIX6) genes. The case report illustrates the wide phenotypic findings associated with these genes along with additional unique findings that previously have not been commonly reported.


Effects of modulators of protein phosphorylation on heme metabolism in human hepatic cells: induction of delta-aminolevulinic synthase mRNA and protein by okadaic acid.
Effects of modulators of protein phosphorylation on delta-aminolevulinic acid (ALA) synthase and heme oxygenase-1 mRNA were analyzed in the human hepatic cell lines Huh-7 and HepG2 using a quantitative RNase protection assay. Okadaic acid was found to induce ALA synthase mRNA in a concentration-dependent fashion in both Huh-7 and HepG2 cells. The EC(50) for induction of ALA synthase mRNA in Huh-7 cells was 13.5 nM, with maximum increases occurring at okadaic acid concentrations of 25-50 nM. The EC(50) for induction of ALA synthase mRNA in HepG2 cells was 35.5 nM, with maximum increases occurring at okadaic acid concentrations of 50 nM. Concentration-dependent induction of ALA synthase mRNA paralleled the increase in ALA synthase protein. Maximum induction of ALA synthase was observed between 5 and 10 h post-treatment in both cell lines. Induction of ALA synthase mRNA in Huh-7 cells, but not HepG2 cells, was associated with an increase in ALA synthase mRNA stability. Okadaic acid also induced heme oxygenase-1 mRNA in both cell lines, but the magnitude of induction was only twofold, and was rapid and transient. Okadaic acid and phorbol 12-myristate 13-acetate significantly decreased heme-mediated induction of heme oxygenase-1 mRNA in both Huh-7 and HepG2 cells. Wortmannin diminished the heme-mediated induction of heme oxygenase-1 mRNA in HepG2 cells, but not Huh-7 cells. These results report a novel property of okadaic acid to affect heme metabolism in human cell lines.


Substitution of an active site valine uncovers a kinetically slow equilibrium between competent and incompetent forms of choline oxidase.
The enzymatic oxidation of choline to glycine betaine is of interest because organisms accumulate glycine betaine intracellularly in response to stress conditions. This is relevant for the genetic engineering of crops with economic interest that do not naturally possess efficient pathways for the synthesis of glycine betaine and for the potential development of drugs that target the glycine betaine biosynthetic pathway in human pathogens. To date, the best characterized choline-oxidizing enzyme is the flavin-dependent choline oxidase from Arthrobacter globiformis, for which structural, mechanistic, and biochemical data are available. Here, we have replaced a hydrophobic residue (Val464) lining the active site cavity close to the N(5) atom of the flavin with threonine or alanine to investigate its role in the reaction of choline oxidation catalyzed by choline oxidase. The reductive half-reactions of the enzyme variants containing Thr464 or Ala464 were investigated using substrate and solvent kinetic isotope effects, solvent viscosity effects, and proton inventories. Replacement of Val464 with threonine or alanine uncovered a kinetically slow equilibrium between a catalytically incompetent form of enzyme and an active species that can efficiently oxidize choline. In both variants, the active form of enzyme shows a decreased rate of hydroxyl proton abstraction from the alcohol substrate, with minimal changes in the subsequent rate of hydride ion transfer to the flavin. This study therefore establishes that a hydrophobic residue not directly participating in catalysis plays important roles in the reaction of choline oxidation catalyzed by choline oxidase.


Contribution of Meat Inspection to the surveillance of poultry health and welfare in the European Union.
In the European Union, Meat Inspection (MI) aims to protect public health by ensuring that minimal hazardous material enters in the food chain. It also contributes to the detection and monitoring of animal diseases and welfare problems but its utility for animal surveillance has been assessed partially for some diseases only. Using the example of poultry production, we propose a complete assessment of MI as a health surveillance system. MI allows a long-term syndromic surveillance of poultry health but its contribution is lowered by a lack of data standardization, analysis and reporting. In addition, the probability of case detection for 20 diseases and welfare conditions was quantified using a scenario tree modelling approach, with input data based on literature and expert opinion. The sensitivity of MI appeared to be very high to detect most of the conditions studied because MI is performed at batch level and applied to a high number of birds per batch.


Characterization of Calcification Events Using Live Optical and Electron Microscopy Techniques in a Marine Tubeworm.
Characterizing the first event of biological production of calcium carbonate requires a combination of microscopy approaches. First, intracellular pH distribution and calcium ions can be observed using live microscopy over time. This allows identification of the life stage and the tissue with the feature of interest for further electron microscopy studies. Life stage and tissues of interest are typically higher in pH and Ca signals. Here, using H. elegans, we present a protocol to characterize the presence of calcium carbonate structures in a biological specimen on the scanning electron microscope (SEM), using energy-dispersive X-ray spectroscopy (EDS) to visualize elemental composition, using electron backscatter diffraction (EBSD) to determine the presence of crystalline structures, and using transmission electron microscopy (TEM) to analyze the composition and structure of the material. In this protocol, a focused ion beam (FIB) is used to isolate samples with dimension suitable for TEM analysis. As FIB is a site specific technique, we demonstrate how information from the previous techniques can be used to identify the region of interest, where Ca signals are highest.


Psychiatric Disorders and Distal 21q Deletion-A Case Report.
Partial deletion of chromosome 21q is a very rare genetic condition with highly variable phenotypic features including heart defects, high or cleft palate, brain malformations (e.g., cerebral atrophy), developmental delay and intellectual disability. So far, there is very limited knowledge about psychiatric disorders and their effective treatment in this special population. To fill this gap, the authors present the case of an initially five-year-old girl with distal deletion (del21q22.2) and comorbid oppositional defiant disorder (main psychiatric diagnosis) covering a period of time of almost four years comprising initial psychological/psychiatric assessment, subsequent treatment with Parent-Child Interaction Therapy (PCIT), and follow-up assessments. Post-intervention results including a 19-month follow-up indicated good overall efficacy of PCIT and high parental satisfaction with the treatment. This case report makes a substantial contribution to enhancing knowledge on psychiatric comorbidity and its effective treatment in patients with terminal 21q deletion. Moreover, it emphasizes the necessity of multidisciplinarity in diagnosis and treatment due to the variety of anomalies associated with 21q deletion. Regular screenings for psychiatric disorders and (if indicated) thorough psychological and psychiatric assessment seem to be reasonable in most affected children, as children with developmental delays are at increased risk of developing psychiatric disorders. As demonstrated with this case report, PCIT seems to be a good choice to effectively reduce disruptive behaviors in young children with partial deletion of chromosome 21q.


Magnetic resonance spectroscopy to study hepatic metabolism in diffuse liver diseases, diabetes and cancer.
This review provides an overview of the current state of the art of magnetic resonance spectroscopy (MRS) in in vivo investigations of diffuse liver disease. So far, MRS of the human liver in vivo has mainly been used as a research tool rather than a clinical tool. The liver is particularly suitable for static and dynamic metabolic studies due to its high metabolic activity. Furthermore, its relatively superficial position allows excellent MRS localization, while its large volume allows detection of signals with relatively low intensity. This review describes the application of MRS to study the metabolic consequences of different conditions including diffuse and chronic liver diseases, congenital diseases, diabetes, and the presence of a distant malignancy on hepatic metabolism. In addition, future prospects of MRS are discussed. It is anticipated that future technical developments such as clinical MRS magnets with higher field strength (3 T) and improved delineation of multi-component signals such as phosphomonoester and phosphodiester using proton decoupling, especially if combined with price reductions for stable isotope tracers, will lead to intensified research into metabolic syndrome, cardiovascular disease, hepato-biliary diseases, as well as non-metastatic liver metabolism in patients with a distant malignant tumor.


Preliminary studies on antihyperglycemic effect of aqueous extract of Brassica nigra (L.) Koch in streptozotocin induced diabetic rats.
In the streptozotocin induced diabetic rats treated separately with aqueous, ethanol, acetone and chloroform extracts of the seeds of B. nigra, the increase in serum glucose value between 0 and 1 hr of glucose tolerance test (GTT) was the least (29 mg/dl) in aqueous extract treated animals while it was 54, 44 and 44 mg/dl with chloroform, acetone and ethanol extracts respectively. In further studies carried out with aqueous extract, the effective dose was found to be 200 mg/kg body weight in GTT. Administration of 200 mg/kg body weight of aqueous extract to diabetic animals daily once for one month brought down fasting serum glucose (FSG) levels while in the untreated group FSG remained at a higher value. In the treated animals the increase in glycosylated hemoglobin (HbA1c) and serum lipids was much less when compared with the levels in untreated diabetic controls. These findings suggest that further studies with the aqueous extract of B. nigra seeds on its antidiabetic activity would be useful.


Hypolipidemic activity of Eclipta prostrata (L.) L. leaf extract in atherogenic diet induced hyperlipidemic rats.
In atherogenic diet induced hyperlipidemic model, the rats receiving treatment with the aqueous extract of the leaves of E. prostrata showed significant reduction in total cholesterol, triglyceride, total protein and elevation in high density lipoprotein cholesterol. The aqueous extract of E. prostrata was found to possess significant hypolipidemic activity. The results also suggest that E. prostrata leaf extract at 100 and 200 mg/kg b.wt. concentrations is an excellent lipid-lowering agent.


Environmentally relevant lead alters nuclear integrity in erythrocytes and generates oxidative stress in liver of Anabas testudineus: Involvement of Nrf2-Keap1 regulation and expression of biomarker genes.
Genotoxic and hepatotoxic effects of lead (Pb) on a freshwater fish, climbing perch (Anabas testudineus) were studied at an environmentally relevant concentration (43.3 ppm). The genotoxic potential of Pb was confirmed by micronucleus study, with increased frequencies of erythrocytic nuclear alterations like lobed, blebbed, notched, fragmented, and micronuclei were observed in erythrocytes in treated groups as compared to control. Inorganic Pb induces oxidative stress which is a consequence of elevated level of Reactive Oxygen Species. Hepatotoxicity was assessed both by the oxidative stress and cellular responses that emerged due to the toxic assault of Pb in the liver, the most important detoxifying organ. Upregulation of xenobiotic metabolizing enzyme like catalase was evident after 15, 30, and 90 days of exposure, and a profound effect was observed on 30th days. The level of lipid peroxidation and reduced glutathione was increased after Pb exposure. Histoarchitectural damages of liver were distinctly evident in treated fish. Western blot analysis confirmed the expressional alterations of stress-responsive marker proteins like Nrf2, Keap1, Hsp70, and Nqo1. Pb exposure resulted in increased expression of Hsp70, Nrf2, and Nqo1, whereas Keap1 was downregulated, suggesting the involvement of Nrf2-Keap1 regulation as a cytoprotective mechanism against Pb toxicity.


MITOCHONDRIAL DNA VARIATION AND EVOLUTION OF THE DEATH VALLEY PUPFISHES (CYPRINODON, CYPRINODONTIDAE).
A phylogenetic analysis of mitochondrial DNA (mtDNA) restriction sites was used to examine the geographic history of the Cyprinodon nevadensis complex of pupfishes, a group of four species (seven extant subspp.) in two endorheic (closed) basins of the Death Valley System in California and Nevada (Owens River Valley and Ash Meadows-Death Valley). The mtDNA results suggest that the group contains mtDNAs from two divergent clades. One such clade is represented by the mtDNAs of the Owens Valley pupfish (C. radiosus) and the existing species in the Colorado River (C. macularius), while the other includes the mtDNAs of the Ash Meadows-Death Valley species (C. nevadensis, C. salinus, and C. diabolis) and a species located much farther to the east (C. fontinalis from the Guzman Basin, Chihuahua, Mexico). These results, together with evidence from other studies, suggest two separate invasions of the Death Valley System by pupfishes carrying phylogenetically divergent mtDNAs. The C. nevadensis complex apparently is either an artificial group or else it is monophyletic and its genetic history includes loss of the original mtDNA in either Owens Valley or Ash Meadows-Death Valley following genetic introgression after an invasion by a pupfish carrying a divergent mtDNA.