Riboflavin deficiency causes protein and DNA damage in HepG2 cells, triggering arrest in G1 phase of the cell cycle.
Eukaryotes convert riboflavin to flavin adenine dinucleotide, which serves as a coenzyme for glutathione reductase and other enzymes. Glutathione reductase mediates the regeneration of reduced glutathione, which plays an important role in scavenging free radicals and reactive oxygen species. Here we tested the hypothesis that riboflavin deficiency decreases glutathione reductase activity in HepG2 liver cells, causing oxidative damage to proteins and DNA, and cell cycle arrest. As a secondary goal, we determined whether riboflavin deficiency is associated with gene expression patterns indicating cell stress. Cells were cultured in riboflavin-deficient and riboflavin-supplemented media for 4 days. Activity of glutathione reductase was not detectable in cells cultured in riboflavin-deficient medium. Riboflavin deficiency was associated with an increase in the abundance of damaged (carbonylated) proteins and with increased incidence of DNA strand breaks. Damage to proteins and DNA was paralleled by increased abundance of the stress-related transcription factor GADD153. Riboflavin-deficient cells arrested in G1 phase of the cell cycle. Moreover, oxidative stress caused by riboflavin deficiency was associated with increased expression of clusters of genes that play roles in cell stress and apoptosis. For example, the abundance of the pro-apoptotic pleiomorphic adenoma gene-like 1 gene was 183% greater in riboflavin-deficient cells compared with riboflavin-sufficient controls. We conclude that riboflavin deficiency is associated with oxidative damage to proteins and DNA in liver cells, leading to cell stress and G1 phase arrest.


Study of the Kuznets environmental curve hypothesis from a global perspective 1960-2019: a semi-parametric panel data proposal.
This work uses parametric and semiparametric panel data analysis methodologies to test the hypothesis of the environmental Kuznets curve, in 186 countries in the period 1960-2019. The main results reveal the acceptance of this hypothesis in the relationships of CO2 emissions (kt) and economic growth (GDP) and urbanization (% population) in the parametric models. Using semiparametric methods, the polynomial relations of fourth degree between CO2 emissions and GDP and of third degree between it and urbanization are verified. The economic policy implications derived from these results seem to indicate the need to continue making efforts in the reduction of CO2 emissions, through greater efforts in innovation and research and development, in search of clean and less polluting energies. The relationship between CO2 and economic growth is a major challenge, in terms of achieving a flattening of this relationship.


Optimal methods for preparation, separation, and determination of radium isotopes in environmental and biological samples.
In recent years, radium has attracted considerable attention primarily because of the rapid increase in unconventional (fracking) drilling technology in the United States and around the world. One of the major radionuclides of interest in unconventional drilling wastes is radium isotopes (224Ra, 226Ra, 228Ra). To access long-term risks associated with radium isotopes entering into the environment, accurate measurements of radium isotopes in environmental and biological samples are crucial. This article reviews many aspects of radium chemistry, which includes recent developments in radiochemical separations methods, advancements in analytical techniques followed by a more detailed discussion on the recent trends in radium determination.


Comparative evaluation of canine cadaver embalming methods for veterinary anatomy education.
Formalin-embalmed cadavers have been extensively used to teach anatomy. Although they ensure the preservation of anatomical structures without microbial contamination, they are considerably rigid and cannot be used to study the joint and muscle movements. Moreover, formalin irritates the eyes and airways and is carcinogenic on chronic exposure. To overcome the disadvantages of formalin-fixed cadavers, we investigated the usefulness of alternative embalming methods using saturated salt solution (SS) and Thiel's solution (TS). We compared the three solutions based on the following parameters: cost of the embalming solution; preservation of anatomical structure, color, flexibility, and texture; and microbial contamination. Convenience of anatomical structure identification and preferences in anatomical laboratory practice were evaluated using questionnaires answered by veterinary undergraduate students. Cost of the embalming solution was the lowest for formalin solution (FS) and most expensive for TS. All cadavers were successfully preserved without significant putrefaction and were useful for teaching veterinary anatomy. Cadavers embalmed with SS or TS were superior in facilitating joint and muscle movement. Compared to FS, the color and texture of muscles and internal organs were similar to those of living animals and there was no irritating and offensive smell in SS and TS cadavers. Students preferred the SS and TS cadavers for their usefulness in identification of anatomical structures, highlighting their usefulness in veterinary anatomy education.


[Biliary surgery using the ligamentum teres. Technique for repairing loss of substance in the common bile duct].
Ligamentum teres was used to repair a total or partial common duct defect in seven cases: 4 cases of biliobiliary stone fistula, 2 cases of hydatid cysto-biliary fistula and one case of iatrogenic common bile duct trauma. Biliary duct reconstruction is possible in the above cases of biliary damage. The short-term and long-term results were satisfactory.


Accuracy of radiation field alignment in clinical practice.
We present an analysis of simulator and portal films of 71 patients. Twenty-five were analyzed retrospectively, 39 prospectively, but without changing routine filming practice, and 7 had daily portal films taken. Treatment-to-treatment variations in anatomy with respect to the field were determined by comparing sequential portal films. The standard deviation of the variations was approximately normally distributed with an average value of 3 mm independent of site and field shaping technique. Discrepancies between the portal and simulator films were greater and depended on the site of treatment. The mean worst-case discrepancy averaged over all sites was 7.7 mm; the lowest value was 3.5 mm in the head and neck region; the highest value was 9.2 mm in the thorax.


Dose of 3-methylcholanthrene enhances vitamin C accumulation and mRNA expression of its transporter in the liver of ODS rats and in HepG2 cells.
Polycyclic aromatic hydrocarbon (PAH) compounds including 3-methylcholanthrene induce harmful reactive intermediates and reactive oxygen species. This study reports the effect of 3-methylcholanthrene on the accumulation of vitamin C and the expression of vitamin C transporters. ODS rats were given l-ascorbic acid daily and intraperitoneal injections of 10 mg 3-methylcholanthrene in total. On day 10, vitamin C concentrations and the expression of vitamin C transporter in the tissues were measured. As a result, the levels of sodium-dependent vitamin C transporter (SVCTs) 1 and the l-ascorbic acid concentration in 3-methylcholanthrene-treated livers and hepatocytes have increased significantly. However, the content of vitamin C in the urine and TBARS in the liver have not changed. These results suggest that the administration of 3-methylcholanthrene elevates the requirement for vitamin C via (SVCTs) 1 due to xenobitics-metabolizing, such as the induction of cytochrome P450 family.


Nucleophilicity and accessibility calculations of alkanolamines: applications to carbon dioxide absorption reactions.
Both nucleophilicities and accessibilities of three alkanolamines [monoethanolamine (MEA), (2-(methylamino)ethanol (MAE), and 2-amino-2-methyl-1-propanol (AMP)] were calculated to predict their reactivities with CO(2). After DFT geometry-optimization calculations, the global, group, and atomic nucleophilicities of each amine were obtained using MP2 quantum mechanical calculations. Only global nucleophilicity matched an experimental pK(a) order (MAE > AMP > MEA). However, it failed to predict the slow rate of the sterically hindered AMP and the order of rate constants, MAE > MEA > AMP. The accessibilities of amines to CO(2) have been calculated by MD simulations by monitoring collisions at the reaction centers: N atoms in amines and C in CO(2). The accessibility results indicate that global nucleophilicity needs quantitative correction for steric effects to predict better reactivities of amines with CO(2).


Prostate radiotherapy in the setting of bilateral hip prostheses: All commonly used photon-based radiation approaches are feasible.
The prevalence of hip prostheses is increasing. Prostate radiation delivery in the setting of hip prostheses is complicated by both imaging artifacts that interfere with volume delineation and dosimetric effects that must be addressed in the planning process. We hypothesized that with specialized planning, any photon-based definitive prostate radiotherapy approach may be utilized in patients with bilateral hip prostheses. Imaging data from sequential patients with prostate cancer and bilateral hip prostheses treated definitively with radiation were retrospectively reviewed. Bimodality imaging was used to define targets and organs at risk (OARs) along with specialized MRI sequences and/or orthopedic metal artifact reduction (OMAR) for MRI and CT artifact suppression, respectively. Multiple VMAT plans were generated for each set of patient images to include three fractionation schemes (conventional, hypofractionated, and SBRT), each with hip avoidance and with simulated normal hip. The ability to meet standard dose constraints was assessed for each plan type. Differences in target and OAR dosing between plans accounting for prosthetic hips via avoidance vs plans with simulated absence of prosthetic hip were also assessed. T-tests were used to compare dosimetric parameters. Ten patients with bilateral hip prostheses were identified, and 6 plans were created for each patient for a total of 60 radiation plans. Prosthetic hip avoidance did not result in failure to meet dose constraints for any patient. Hip avoidance resulted in minimal increases in high dose to the rectum and bladder (increases in mean V80%, V90%, and V95% ranged from 0.1% to 2.4%). Larger increases were seen at lower dose levels, with rectal V50% significantly increased in all three plan types with hip avoidance (conventional: 26.0% [standard deviation, SD 13.9] vs 16.9% [SD 10.2, p = 0.003]; hypofractionation: 26.4% [SD 13.3] vs 17.1% [SD 10.1, p = 0.002]; SBRT: 18.3% [SD 10.7] vs 10.5% [SD 6.9, p = 0.008]). Similarly, hip avoidance resulted in increases in bladder V50% to 31.7% (SD 16.8) vs 23.3% (SD 14.0, p = 0.001), 31.3% (SD 17.0) vs 23.3% (SD 13.8, p = 0.002), and 22.7% (SD 12.3) vs 16.5% (SD 12.6, p < 0.001) for conventional, hypofractionated, and SBRT plans, respectively. Hydrogel spacer resulted in reductions in rectal dose. For example, V70% for hip avoidance plans decreased with spacer presence to 8.3% (SD 6.7) vs 21.1% (SD 5.8, p = 0.021), 8.6% (SD 6.5) vs 21% (SD 5.7, p = 0.022), and 3.7% (SD 3.2) vs 15% (SD 8.2, p = 0.010) for conventional, hypofractionated, and SBRT plans, respectively. Any photon-based definitive prostate radiotherapy approach can be used with bimodality imaging for target and OAR definition and planning techniques to avoid dose attenuation effects of hip prostheses. Hydrogel spacer is a useful adjunct.


A transferable "resistance factor" from in vitro cultured MDMS-resistant Yoshida sarcoma cells.
Cells of the methylene dimethanesulphonate-(MDMS)-resistant Yoshida sarcoma cell line contain a low molecular weight "resistance factor" which is present in the culture medium of these cells and may be utilized by MDMS-sensitive Yoshida sarcoma cells either by co-culturing the two cell lines or by culturing the MDMS-sensitive Yoshida cells in a medium containing 20% used medium of MDMS-resistant Yoshida cells or in the presence of dialysed medium from resistant cells. The "resistance factor" does not inactivate the drug itself or its metabolites, and it has no influence on the sensitivity of the cells if added after MDMS treatment. Twenty-four hours seems to be enough time for the transfer of the resistance factor, but its effect on whole populations decreases within 24 hours of ceasing the supply. The relationship between these findings and the known phenomena of metabolic co-operation are discussed.


Linac-based radiosurgery of cerebral melanoma metastases. Analysis of 122 metastases treated in 64 patients.
Stereotactic radiosurgery is an alternative option to neurosurgical excision in the management of patients with brain metastases. We retrospectively analyzed patients with brain metastases of malignant melanoma who were treated at our institution for outcome and prognostic factors. 64 patients with 122 cerebral metastases were treated with stereotactic radiosurgery between 1986 and 2000. Twelve patients (19%) showed neurologic symptoms at the time of treatment, and 46 patients (72%) had extracerebral tumor manifestation at that time. The median dose to the 80% isodose line, prescribed to encompass the tumor margin, was 20 Gy (range, 15-22 Gy). Neurologic symptoms improved in five of twelve symptomatic patients. 41 patients remained asymptomatic or unchanged in their neurologic symptoms. Only five patients (8%) temporarily worsened neurologically after therapy despite no signs of tumor progression. With a mean follow-up time of 9.4 months, actuarial local control was 81% after 1 year. There was a statistically significant dose and size dependency of local tumor control. Median actuarial survival after treatment was 10.6 months. Patients without extracerebral tumor manifestation showed a superior survival (p = 0.04). Despite high local tumor control rates, the prognosis of patients with cerebral metastases of malignant melanoma remains poor. Stereotactic radiosurgery has the potential of stabilizing or improving neurologic symptoms in these patients in a palliative setting.


Dopamine release within forebrain sites perfused with tetrahydroisoquinolines or tryptoline in the rat.
The localized effect of amine-aldehyde condensation products on the endogenous release of dopamine (DA) was examined in dopaminergic-rich areas of the brain. In the unanesthetized rat, dopaminergic stores were labeled by 14C-dopamine (1.0 microCi) microinjected in a volume of 1.0 microliters into the caudate nucleus, tuberculum olfactorium, nucleus accumbens or other contiguous sites within the ventral forebrain. Subsequently, each labeled site was locally perfused with an artificial CSF, by means of push-pull cannulae, at a rate of 25.0 microliters per min. Two tetrahydroisoquinolines, tetrahydropapaveroline and 4,6,7-TH-tetrahydroisoquinoline, as well as the tetrahydro-beta-carboline, tryptoline, significantly enhanced the release of 14C-DA when each alkaloid metabolite was perfused in a concentration of 250 nanograms per min. Tetrahydroisoquinoline was without effect on the local kinetics of 14C-DA, whereas salsolinol had an intermediary action on DA release. These findings suggest that an alkaloid conjugate when given in the cerebral ventricle could exert its central effects by way of a dopaminergic mechanism in the telencephalon.


Wound healing by transplantation of mesenchymal stromal cells loaded on polyethylene terephthalate scaffold: Implications for skin injury treatment.
Several clinical studies have shown that cellular therapy based on mesenchymal stromal cells (MSCs) transplantation may accelerate wound healing. One major challenge is the delivery system used for MSCs transplantation. In this work, we evaluated the capacity of a scaffold based on polyethylene terephthalate (PET) to maintain the viability and biological functions of MSCs, in vitro. We examined the capacity of MSCs loaded on PET (MSCs/PET) to induce wound healing in an experimental model of full-thickness wound. Human MSCs were seeded and cultured on PET membranes at 37 °C for 48 h. Adhesion, viability, proliferation, migration, multipotential differentiation and chemokine production were evaluated in cultures of MSCs/PET. The possible therapeutic effect of MSCs/PET on the re-epithelialization of full thickness wounds was examined at day 3 post-wounding in C57BL/6 mice. Histological and immunohistochemical (IH) studies were performed to evaluate wound re-epithelialization and the presence of epithelial progenitor cells (EPC). As controls, wounds without treatment or treated with PET were established. We observed MSCs adhered to PET membranes and maintained their viability, proliferation and migration. They preserved their multipotential capacity of differentiation and ability of chemokine production. MSCs/PET implants promoted an accelerated wound re-epithelialization, after three days post-wounding. It was associated with the presence of EPC Lgr6+ and K6+. Our results show that MSCs/PET implants induce a rapid re-epithelialization of deep- and full-thickness wounds. MSCs/PET implants constitute a potential clinical therapy for treating cutaneous wounds.


Photodynamic Therapy for the Treatment of Complex Anal Fistula.
To validate and analyze the results of intralesional photodynamic therapy in the treatment of complex anal fistula. This prospective multicentric observational study enrolled patients treated for complex anal fistula who underwent intralesional photodynamic therapy (i-PDT). The included patients were treated from January 2016 to December 2018 with a minimum follow-up of 1 year to evaluate recurrence, continence and postoperative morbidity. Intralesional 5-aminolevulinic acid (ALA) gel (2%) was injected directly into the fistula. The internal and external orifices were closed. After an incubation period of 2 hours, the fistula was irradiated using an optical fiber connected to a red laser (Multidiode 630 PDT) operating at 1 W/cm for 3 minutes (180 J). In total, 49 patients were included (61.2% male). The mean age was 48 years, and the mean duration of fistula was 13 months. Of the fistulas included, 75.5% were medium transphincteric, and 24.5% were high transphincteric. The median fistula length was 4 ± 1,14 cm (range: 3-5). A total of 41 patients (83.7%) had a previous history of fistula surgery. Preoperatively, some degree of anal incontinence was found in 5 patients (10.2%). No center reported any other procedure-related complications intraoperatively. Phototoxicity was found in one patient. In the first 48 hours after the procedure, fever was reported in 2 patients (4%). At the end of follow-up, total healing was observed in 32/49 patients (65.3%). No patient reported new incontinence postoperatively. i-PDT could be considered a good choice in patients with complex anal fistulas to avoid surgery and its complications. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.


Experiences of adolescents living with Silver-Russell syndrome.
The psychosocial impact of growing up with Silver-Russell syndrome (SRS), characterised by growth failure and short stature in adulthood, has been explored in adults; however, there are no accounts of contemporary lived experience in adolescents. Such data could inform current healthcare guidance and transition to adult services. We aimed to explore the lived experience of adolescents with SRS. In-depth, semi-structured interviews were conducted between January 2015 and October 2016 with a sample of eight adolescents aged 13-18 (five girls) with genetically confirmed SRS from the UK. Qualitative interviews were transcribed and coded to identify similarities and differences using thematic analysis; codes were then grouped to form overarching themes. We identified four themes from the interview data: (1) the psychosocial challenges of feeling and looking different; (2) pain, disability and fatigue; (3) anticipated stigma; and (4) building resilience and acceptance. Despite adolescents accepting SRS in their lives, they described ongoing psychosocial challenges and anticipated greater problems to come, such as stigma from prospective employers. Adolescents with SRS may experience psychosocial difficulties from as young as 10 years old related to feeling and looking different; pain, disability and fatigue; anticipated stigma; and future challenges around employment. We discuss these findings in relation to recommendations for the care of adolescents with SRS to prepare them for adult life.


crAssphage as a human molecular marker to evaluate temporal and spatial variability in faecal contamination of urban marine bathing waters.
Bathing water quality may be negatively impacted by diffuse pollution arising from urban and agricultural activities and wildlife, it is therefore important to be able to differentiate between biological and geographical sources of faecal pollution. crAssphage was recently described as a novel human-associated microbial source tracking marker. This study aimed to evaluate the performance of the crAssphage marker in designated bathing waters. The sensitivity and specificity of the crAss_2 marker was evaluated using faecal samples from herring gulls, dogs, sewage and a stream impacted by human pollution (n = 80), which showed that all human impacted samples tested positive for the marker while none of the animal samples did. The crAss_2 marker was field tested in an urban marine bathing water close to the discharge point of human impacted streams. In addition, the bathing water is affected by dog and gull fouling. Analysis of water samples taken at the compliance point every 30 min during a tidal cycle following a rain event showed that the crAss_2 and HF183 markers performed equally well (Spearman correlation ρ = 0.84). The levels of these marker and faecal indicators (Escherichia coli, intestinal enterococci, somatic coliphages) varied by up to 2.5 log10 during the day. Analysis of a high-tide transect perpendicular to the shoreline revealed high levels of localised faecal contamination 1 km offshore, with a concomitant spike in the gull marker. In contrast, both the crAss_2 and HF183 markers remained at a constant level, showing that human faecal contamination is homogenously distributed, while gull pollution is localised. Performance of the crAss_2 and HF183 assay was further evaluated in bimonthly compliance point samples over an 18-month period. The co-occurrence between the crAss_2 and HF183 markers in compliance sampling was 76%. A combination of both markers should be applied in low pollution impacted environments to obtain a high confidence level.


Reproducibility and efficiency of serum-derived exosome extraction methods.
Exosomes are emerging as a source of biomarkers with putative prognostic and diagnostic value. However, little is known about the efficiency, reproducibility and reliability of the protocols routinely used to quantify exosomes in the human serum. We used increasing amounts of the same serum sample to isolate exosomes using two different methods: ultracentrifugation onto a sucrose cushion and ExoQuick™. Quantitative analysis of serum-derived exosomes was performed by determining protein concentration (BCA assay) and the number of nanoparticles (Nanosight™ technology). Exosome quality was assessed by Coomassie staining and Western blotting for CD9, LAMP2 exosomal markers and a negative marker Grp94. Correlation between serum volume and the number of isolated exosomes is significant for both methods when exosomes are quantified using protein concentration. However, when the number of nanoparticles is used to quantify exosomes, ExoQuick™ is the only reproducible and efficient method. CD9, LAMP2 and Grp94 exosomal markers are equivalently expressed in both methods. However, exosomes isolated using ultracentrifuge method are strongly contaminated with albumin and IgG. ExoQuick™ is an efficient and reproducible method to isolate exosomes for quantitative studies, whereas ultracentrifugation is not. Moreover, high albumin contamination of ultracentrifuged-derived exosomes impairs the use of protein concentration as a mean to quantify serum-derived exosomes.


Perceptions of care coordination in a population-based sample of diverse breast cancer patients.
To identify factors associated with perceptions of care coordination in a diverse sample of breast cancer patients. Breast cancer patients reported to the metropolitan SEER registries of Detroit or Los Angeles from 6/05 to 2/07 were surveyed after diagnosis (N=2268, RR=72.4%). Outcomes were two dichotomous measures reflecting patient appraisal of care coordination during their treatment experience. Primary independent variables were race/ethnicity (white, African American, Latina-high acculturated, Latina-low acculturated) and health literacy (low, moderate, high). Logistic regression was used to evaluate factors associated with both measures of care coordination. 2148 subjects were included in the analytic dataset. 16.4% of women perceived low care coordination and 12.5% reported low satisfaction. Race/ethnicity was not significantly associated with care coordination. Women with low subjective health literacy were 3-4 times as likely as those with high health literacy to perceive low care coordination and low satisfaction with care coordination (OR=3.88; 95% CI: 2.78-5.41; OR=3.19 95% CI: 2.25-4.52, respectively). Many breast cancer patients positively appraised their care coordination, but patients with low health literacy perceived low care coordination. Providers should be aware of the health literacy deficits that may contribute to their patients' attitudes towards their breast cancer care coordination.


How to treat lumbar disc herniation in pregnancy? A systematic review on current standards.
In this systematic review, we aim to illustrate the current and safe concepts in the assessment, diagnosis and management of herniated lumbar disc (HLD) during pregnancy. A systematic review and reporting on the diagnosis, treatment and clinical results of HLD during pregnancy is performed. The MRI represents the first level and safest diagnostic tool for pregnant women affected by spinal problems allowing for a noninvasive and detailed radiological examination of the spine. The initial management of pregnant women affected by HLD is conservative, and primarily aimed to pain therapy. Whenever radicular pain and progressive neurological deficits unresponsive to medical management occur, surgery should be considered. Few case reports regarding the operative management of HLD in pregnant women have been published up to date. Laminectomy and/or microdiscectomy represent the classical and most commonly used techniques that can be safely performed without affecting pregnancy, delivery, or baby's health. Endoscopic discectomy may be an alternative. The most adequate timing and surgical position are chosen based on to the fetal gestational age and site of the pathology. Surgical treatments during pregnancy impose multiple medical and ethical problems. Timely diagnosis by MRI, careful clinical evaluation, and surgical treatment represent safe and effective procedures. Ongoing evolution of surgical, anesthesiological and obstetrical procedures results in favorable outcomes. However, interdisciplinary management and a wide knowledge of pregnancy-related pathologies are crucial for the best outcome for both mother and child.


The cellular content of non Hodgkin lymphomas: a comprehensive analysis using monoclonal antibodies and other surface marker techniques.
Five samples of tonsil, 10 reactive lymph nodes and 65 consecutive cases of non-Hodgkin lymphoma (NHL) were evaluated in suspension phenotyping with the monoclonal antibodies alpha Leu-I, alpha Leu-2a, alpha Leu-3a, OKT1, OKT3, OKT4, OKT6, OKT8, W6/32, 26/114, DA-2, 2DI, J5, AN51 and OKT9 together with conventional surface marking by rosetting (E, Fc gamma, Fc mu, C3b, C3d) and staining for surface and cytoplasmic immunoglobulin (SIg, CyIg) heavy and light chain classes. The results confirm the reproductability and specificity of staining with monoclonal antibodies against T cells and T cells subsets. Evidence is presented for reactivity of alpha Leu-I antibody with SIg positive and Ia positive cells in some lymphomas (centroblastic centrocytic, lymphocytic and immunoblastic), and 2 cases showed evidence of marking with OKT3 on SIg positive cells in T cell predominant immunoblastic lymphoma. Lymphoblastic lymphomas of T cell type expressed the marker OKT6. On the basis of these results criteria for the diagnosis of T cell lymphoma are suggested. The monoclonal antibody J5, reactive with C-ALL antigen, showed variable positivity, occasionally strong in B cells in cases of centroblastic and centrocytic follicular lymphoma. Proportions of cells staining with the monoclonal antibody OKT9 showed a correlation between levels of cellular expression of transferrin (trf) receptor and the histological grade of malignancy, OKT9+ cells being elevated in high grade lymphomas, and in some cases of transforming lymphoma of low grade histological class. These results are discussed and indicate the advantage of employing a wide range of defined monoclonal reagents in the phenotypic evaluation of NHL.


Sensory processing measure-HK Chinese version: psychometric properties and pattern of response across environments.
This study aimed to examine the psychometric properties of the Sensory Processing Measure-Hong Kong Chinese version (SPM-HKC), and to study the pattern of behavioral response of children towards sensory events across home and school settings. The two major forms of the SPM, Home Form and Main Classroom Form, were translated into Chinese in this study. The content validity of the SPM-HKC was reviewed by 20 expert panel members. A total of 547 typically developing children and 140 children with autistic spectrum disorder (ASD) were recruited for the field test on its reliability and validity. The findings of this study showed that the SPM-HKC was a valid and reliable tool in the screening for sensory processing difficulty of children aged 5-12 among the Chinese populations. But the correlation between the Home Form and the Main Classroom Form was low. It is recommended to use separate forms and norms to measure the performance of children across the home and school environments for more thorough understanding of difficulty in encountering daily sensory events.


How to determine institutional needs and requirements for a trauma registry.
As institutions become interested in trauma registries, they find an overwhelming variety of decisions to be considered. A specific process can be used to simplify the procedure and to ensure a successful choice of a trauma registry.


Replication in Saccharomyces cerevisiae of plasmid pBR313 carrying DNA from the yeast trpl region.
Plasmid pBR313 carrying a 1.4 kb EcoRI fragment from the yeast TRP1 region (designated pLC544) is capable of transforming yeast trp1 mutants to Trp+ at high frequency (10(3)--10(4) transformants/micrograms DNA). Transformation can be achieved either by using purified plasmid DNA or by fusion of yeast spheroplasts with partially lysed Escherichia coli [pLC544] protoplast preparations. The Trp+ yeast transformants are highly unstable, segregating Trp- cells at frequencies of 0.18 per cell per generation (haploids) and 0.056 per cell per generation (diploids) in media containing tryptophan. Plasmid pLC544 replicates autonomously in the nucleus of yeast cells and segregation of Trp-cells is associated with the complete loss of plasmid sequences. In genetic crosses, pLC544 is randomly assorted during meiosis and is carried unchanged through the mating process into haploid recombinants.


Macronutrient selection by foraging rats.
Rats were studied in a laboratory simulation of foraging for separate sources of carbohydrate, fat, and protein. Fixed ratios of bar-presses were required to search for a cue signalling the availability of a meal and to procure each meal. In the first phase, the costs of procuring all three foods were raised equally for three rats. Meal frequency declined and meal size increased for all three foods. Protein opportunities were accepted more frequently than those for other foods, and the proportion of calories taken from protein remained low and constant. Two rats were studied in the remaining three phases. When protein meal cost was raised while the costs of energy foods were low, protein intake was gradually reduced and at the highest cost was dropped from the diet. There were individual differences in the preferred source of energy calories; when each animal's preferred energy source was made more costly, these preferences persisted over the range of costs studied in one rat, but shifted abruptly in the other rat. Finally, when search costs were raised, the animals became less selective in accepting meal opportunities and composing their diets. These results suggest that diet selection in the rat is flexible and responsive to changes in the costs of acquiring food. This ability is consistent with the foraging behaviors required by omnivorous generalized feeders.


Interspecific abundance-occupancy relations along estuarine gradients.
Do interspecific abundance-occupancy (A-O) relationships vary systematically along environmental gradients? A-O relationships of macrobenthic assemblages of seagrass and adjacent bare-sediment were compared at series of sites along two types of estuarine gradient; the longitudinal one of the main axial channel, and a transverse one from that channel into progressively smaller channels and creeks enclosed within fringing saltmarsh. Three general features emerged: A-O regression slopes were remarkably uniform across all the disparate assemblages and sites (1.04 ± 0.08); values of R2 were very high (≈0.96); and where small but significant differences between sites occurred, they were associated with the stronger gradients (i.e. those with the larger differentials in abundance and smaller compositional similarities). In two such cases, slope values were correlated with position along the gradient (although only at the margin of statistical significance). Variation in A-O relationships along estuarine gradients does clearly occur, but their constancy leaves the stronger impression.


Purification of Recombinant Human 6His-FAD Synthase (Isoform 2) and Quantitation of FAD/Protein Monomer Ratio by UV-Vis Spectra.
Here we describe a protocol for a one-step purification of a soluble form of human FAD synthase (isoform 2; hFADS2), overexpressed as a 6-His-tagged fusion protein in Escherichia coli, with a yield of about 15 mg from 1 L of transformed bacterial culture.Following a desalting procedure, the protein is obtained in its FAD-bound form (about 0.8 molecules of FAD per 1 protein monomer). A simple method is also proposed here, for the rapid estimation of the [FAD ]/[protein monomer] ratio, starting from the typical flavoprotein spectrum of the purified protein fraction.The procedure described gives the protein at a quite high grade of purity (about 95%) and in its bifunctional (184.108.40.206/184.108.40.206) enzymatically active form, useful for further kinetical and molecular characterization.


Lactococcus intestinalis sp. nov., a new lactic acid bacterium isolated from intestinal contents in Alzheimer's disease mice.
A gram-positive, facultatively anaerobic, and coccoid or ovoid-shaped bacterium designated M2458T was isolated from the intestinal contents of APPswe/PSΔE9 mouse model of Alzheimer's disease. With the polyphasic approach, the taxonomic position of the novel isolate was confirmed. Strain M2458T grew well at 37 °C on YCFA agar. Strain M2458T belongs to the family Streptococcaceae and class Bacilli, and it is closed to Lactococcus formosensis NBRC 109475T (97.59% sequence similarity) according to its 16S rRNA gene sequence. In a comparison of two housekeeping genes, rpoA and rpoB, strain M2458T was found to be well separated from Lactococcus formosensis NBRC 109475T. On the basis of whole genome sequences, the DNA G+C content was 38.29 mol%. The phylogentic analysis of the whole genome showed that a different branch was clearly formed in the phylogenetic tree of strain M2458T compared to other strains in the genus Lactococcus. A total of eight genes in strain M2458T are involved in the 'neurodegenerative disease' pathway, which involves an annotated protein (glyceraldehyde 3-phosphate dehydrogenase) involved in Alzheimer's disease. In terms of average nucleotide identity and digital DNA-DNA hybridization, strain M2458T was identified as a novel species of the genus Lacococcus. The major fatty acids (> 10% of the total fatty acids) were C18:1ω9c (39.68%), C16:0 (13.26%) and C18:1ω7c (11.52%). The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and an unidentified phospholipid. As a result of its genotypic and phenotypic characteristics, strain M2458T was considered to be a new species within the genus Lactococcus; the name Lactococcus intestinalis sp. nov. has been proposed, with type strain M2458T (=JCM 35706 = CGMCC 1.60066).


Exercise training fails to prevent glucocorticoid-induced muscle alterations in young growing rats.
The aim of this study was to determine the impact of chronic treatment for 8 weeks with hydrocortisone acetate (5 mg kg-1 day-1) on skeletal muscles, and to evaluate whether sprint training can prevent glucocorticoid-induced muscle atrophy better than endurance training. Biochemical, histological and contractile properties were employed to determine the influence of this steroid on skeletal musculature, and the results were compared to pair-weight animals to take into account the influence of corticoids on growth rate. It was found that hydrocortisone acetate treatment results in a stunted growth, adrenal atrophy and depressed plasma corticosterone levels. Mild corticoid-induced losses of muscle mass and protein content (9%-13%) were observed in fast-twitch skeletal muscles. It appeared that the impact of corticoids is strictly directed toward type IIb fibres, which displayed a 12%-18% reduction in cross-sectional areas. No alterations occurred in plantaris contractile speed or tensions properties. Neither endurance training (30 m/min; 90 min/day; 5 days/week) nor sprint training (60 m/min; 15 min/day; 5 days/week) for 8 weeks was able to counteract the effects of corticoids. These data suggest that increased contractile activity, as induced by treadmill running, is not sufficient to counteract the muscular effects of glucocorticoids when administered at a dose of 5 mg kg-1 day-1.


Malingering in children: fibs and faking.
Research has established that children can make efforts to deceive others and that malingering or underperformance in psychiatric and psychological evaluations is common. Clinicians often resist the idea that children can successfully fake mental disorders and formal assessment for malingering is rare in clinical practice. The author suggests that screening tests be performed during the initial evaluation of all children to identify deceptive behavior. Children who behave in a suspect fashion and children who have known motivations to present as more pathologic than they are should be formally assessed with psychological techniques to rule out the presence of malingering.


Failure of monocytes of trauma patients to convert to immature dendritic cells is related to preferential macrophage-colony-stimulating factor-driven macrophage differentiation.
Following trauma, increased inflammatory monokine activation and depressed APC function can occur simultaneously. These contradictory monocyte (Mphi) dysfunctions could result if postinjury Mphi differentiation preferentially favored inflammatory macrophage (Mac) differentiation over development into the most potent APC, dendritic cells (DC). In this report, Mphi of trauma patients with a depressed MLR induction capacity are, for the first time, shown to be unable to differentiate in vitro to immature CD1a(+) DC under the influence of GM-CSF and IL-4. Trauma patient Mphi that retained MLR-inducing capacity had a nonsignificant reduction in DC differentiation capacity. Only patient Mphi populations with depressed differentiation to immature DC (iDC) demonstrated depressed IL-12 and IL-15 production and a continued reduced MLR induction capacity. Neither increased IL-10 production nor decreased CD11c(+) DC precursor numbers correlated with depressed Mphi-to-DC differentiation. Instead, these patients' APC-dysfunctional Mphi populations had increased expression of inflammatory Mac phenotypes (CD64(+), CD86(low), HLA-DR(low)) and up-regulated secretion of M-CSF. M-CSF combined with IL-6 inhibits Mphi-to-iDC differentiation and promotes Mphi-to-Mac differentiation by down-regulating GM-CSFR expression and increasing DC apoptosis. Both depressed GM-CSFR expression and increased Mphi iDC apoptosis, as well as increased expression of CD126 (IL-6R) and CD115 (M-CSFR), were detected in APC-defective patient Mphi. In vitro addition of anti-M-CSF enhanced the IL-4 plus GM-CSF-induced Mphi-to-DC differentiation of these patients. This suggests that, in trauma patients, enhanced Mphi-to-Mac differentiation with concomitant inhibited iDC development is partially due to increased circulating Mphi sensitivity to and production of M-CSF and contributes to postinjury immunoaberrations.


Regioselective hydrations of 1-aryl-3-en-1-ynes using gold and platinum catalysts: selective production of 2-en-1-ones and 3-en-1-ones.
Regiocontrolled hydrations of 1-aryl-3-en-1-ynes have been accomplished with IPrAuOTf and PtCl2/CO to yield 3-en-1-ones and 2-en-1-ones efficiently; our experimental data indicates that the sizes of catalysts play an important role.


Filamentous phage infection-mediated gene expression: construction and propagation of the gIII deletion mutant helper phage R408d3.
We describe the use of transcriptional fusions to the phage shock protein (psp) promoter. These fusions are expressed only when cells are infected by filamentous phage. In an application, the psp promoter was fused to the protein coding part of filamentous phage gene III (gIII). Protein III (pIII) is needed to complement mutant f1 phage containing a deletion of gIII, but its synthesis also renders cells resistant to infection. By inducing pIII production from psp-gIII only in the cells that are already infected with phage, it was possible to obtain plaques from phage in which gIII had been completely deleted. gIII was deleted from two helper phages: R408 and VCSM13, which were then propagated on cells containing the psp-gIII fusion. These two phages were tested for use in a phage display method that requires generation of noninfectious, phagemid-containing virion-like particles. Both helpers worked, but R408d3 was superior to VCSM13d3, because it generated about 1800-times fewer background infectious particles.


Production of volatile compounds by wild-type yeasts in a natural olive-derived culture medium.
The production of volatile compounds in naturally fermented green table olives from Manzanilla cultivar was investigated. A total of 62 volatile compounds were detected after 24 weeks of fermentation. To clarify the contribution of yeasts to the formation of these compounds, such microorganisms were isolated from the corresponding fermenting brines. Five major yeast strains were identified: Nakazawaea molendinolei NC168.1, Zygotorulaspora mrakii NC168.2, Pichia manshurica NC168.3, Candida adriatica NC168.4, and Candida boidinii NC168.5. When these yeasts were grown as pure cultures in an olive-derived culture medium, for 7 days at 25 °C, the number of volatiles produced ranged from 22 (P. manshurica NC168.3) to 60 (C. adriatica NC168.4). Contribution of each yeast strain to the qualitative volatile profile of fermenting brines ranged from 19% (P. manshurica NC168.3) to 48% (Z. mrakii NC168.2 and C. adriatica NC168.4). It was concluded that C. adriatica NC168.4 presented the best aromatic profile, being a solid candidate to be part of a novel starter culture to enhance the organoleptic properties of naturally fermented green table olives.


Oregano Oil Combined with Macleaya Cordata Oral Solution Improves the Growth Performance and Immune Response of Broilers.
The abuse of AGPs in animal husbandry has led to severe problems such as drug resistance and ecological, and environmental destruction, which seriously threaten human health and public health security. In recent years, extracts of oregano oil and macleaya cordata have become a hot spot in the research and application of AGP substitutes for their safety and high efficiency. This study is the first to report the effect of oregano oil combined with macleaya cordata oral solution on broiler growth performance. A total of 960 one-day-old broiler chickens were randomly divided into four treatment groups (240 chickens per group). Each treatment group was divided into six replicate groups (40 birds per replicate group). There were four groups in this study: the solvent control group, the oregano essential oil combined with macleaya cordata extract oral solution group (OS group), the oregano essential oil oral solution group (OEO group), and the macleaya cordata extract oral solution group (MCE group). Two chickens from each replicate group were collected and mixed into a composite sample. Six composite samples were obtained for each treatment group. The results showed that the oregano oil combined with macleaya cordata oral solution significantly improved the growth performance of broiler chickens. At the same time, serum biochemical indices, serum antioxidant indices, serum immune indices, serum cytokines, and intestinal morphology were significantly improved by the OS group. This study shows that oregano oil combined with macleaya cordata oral solution has substantial potential to be an alternative to AGPs for broilers.


A technique for preparing protein gradients on polymeric surfaces: effects on PC12 pheochromocytoma cells.
A technique for preparing micropatterns and gradients of proteins on polymeric substrates has been developed in this work. Peroxides were generated on the substrate surface by UV preirradiation and they initiated graft polymerization of acrylic acid (AA) onto the surface upon a second UV irradiation. Micropatterns and gradients of poly(acrylic acid) (PAA) were formed when the substrate was placed under or moved with respect to a photomask during UV preirradiation. Protein micropatterns and gradients were fabricated on the surface by covalently linking to the carboxyl groups on PAA chains. To test cell response to the protein gradient surfaces, PC12 pheochromocytoma cells were cultured on laminin-bound substrates in serum-free medium supplemented with nerve growth factor (NGF). It is found that both the attachment and neurite outgrowth behaviors of PC12 cells were dependent on the surface laminin density. However, the unreacted carboxyl groups on the polymer surface negatively affected PC12 cells. This weakened the positive influence from laminin.


Hierarchical Temporal Structuring of Speech: A Multiscale, Multimodal Framework to Inform the Assessment and Management of Neuromotor Speech Disorder.
Hierarchical temporal structuring of speech is the key to multiscale linguistic information transfer toward effective communication. This study investigated and linked the hierarchical temporal cues of the kinematic and acoustic modalities of natural, unscripted speech in neurologically healthy and impaired speakers. Thirteen individuals with amyotrophic lateral sclerosis (ALS) and 10 age-matched healthy controls performed a story-telling task. The hierarchical temporal structure of the speech stimulus was measured by (a) 26 articulatory-kinematic features characterizing the depth, phase synchronization, and coherence of temporal modulation of the tongue tip, tongue body, lower lip, and jaw, at three hierarchically nested timescales corresponding to prosodic stress, syllables, and onset-rime/phonemes, and (b) 25 acoustic features characterizing the parallel aspects of temporal modulation of five critical-spectral-band envelopes. All features were compared between groups. For each aspect of temporal modulation, the contributions of all articulatory features to the parallel acoustic features were evaluated by group. Generally consistent disease impacts were identified on the articulatory and acoustic features, manifested by reduced modulation depths of most articulators and critical-spectral-band envelopes, primarily at the timescales of syllables and onset-rime/phonemes. For healthy speakers, the strongest articulatory-acoustic relationships were found for (a) jaw and lip, in modulating stress timing, and (b) tongue tip, in modulating the timing relation between onset-rime/phonemes and syllables. For speakers with ALS, the tongue body, tongue tip, and jaw all showed the greatest contributions to modulating syllable timing. The observed disease impacts likely reflect reduced entrainment of speech motor activities to finer-grained linguistic events, presumably due to the dynamic constraints of the neuromuscular system. To accommodate these restrictions, speakers with ALS appear to use their residual articulatory motor capacities to accentuate and convey the perceptually most salient temporal cues underpinned by the syllable-centric parsing mechanism. This adaptive strategy has potential implications in managing neuromotor speech disorders.


Vocal symptomatology of postoperative dysphonia.
Vocal symptomatology of 24 patients was assessed following the removal of polyps (12 cases) and polypoid degeneration (12 cases). In addition to fundamental frequency, hoarseness type was measured using the spectrographic system developed by Yanagihara which assesses harmonic structure of phonemes relative to noise components. Mean vocal frequency (155Hz) for the polypoid degeneration group was significantly lower than mean vocal frequency for the polyp group (186Hz). There was no difference between the two groups in mean spectrographic hoarseness type. Analysis of data suggested two subgroups of postoperative dysphonia based upon vocal symptomatology: Type A--little or no hoarseness and Type B--severe hoarseness. Neither preoperative nor postoperative conditions of the vocal cords were related to the severity of hoarseness demonstrated by Type B dysphonics. Illustrative cases are presented. One possible explanation for severe postoperative dysphonia is developed.


Foxp3 exploits a pre-existent enhancer landscape for regulatory T cell lineage specification.
Regulatory T (Treg) cells, whose identity and function are defined by the transcription factor Foxp3, are indispensable for immune homeostasis. It is unclear whether Foxp3 exerts its Treg lineage specification function through active modification of the chromatin landscape and establishment of new enhancers or by exploiting a pre-existing enhancer landscape. Analysis of the chromatin accessibility of Foxp3-bound enhancers in Treg and Foxp3-negative T cells showed that Foxp3 was bound overwhelmingly to preaccessible enhancers occupied by its cofactors in precursor cells or a structurally related predecessor. Furthermore, the bulk of Foxp3-bound Treg cell enhancers lacking in Foxp3(-) CD4(+) cells became accessible upon T cell receptor activation prior to Foxp3 expression, and only a small subset associated with several functionally important genes were exclusively Treg cell specific. Thus, in a late cellular differentiation process, Foxp3 defines Treg cell functionality in an "opportunistic" manner by largely exploiting the preformed enhancer network instead of establishing a new enhancer landscape.


Contribution of psychiatric diagnoses to extent of opioid prescription in the first year post-head and neck cancer diagnosis: A longitudinal study.
The purpose of this study was to determine, within the first-year post-head and neck cancer (HNC) diagnosis, the contribution of past and upon HNC psychiatric diagnoses (ie, substance use disorder, major depressive disorder, and anxiety disorder) to the extent (ie, cumulated dose) of opioid prescription. Prospective longitudinal study of 223 consecutive adults (on 313 approached; 72% participation) newly diagnosed (<2 weeks) with a first occurrence of primary HNC, including Structured Clinical Interviews for DSM-IV disorders, validated psychometric measures, and medical chart reviews. Opioid doses were translated into standardized morphine milligram equivalents (MME) using CDC guidelines. A model of variables was tested using multiple linear regression. Fifty-five percent (123/223) of patients received opioids at some point during the first 12 months post-HNC diagnosis, 37.7% (84/223) upon HNC diagnosis (pre-treatment), 40.8% (91/223) during treatments, and 31.4% (70/223) post-treatment. The multiple linear regression indicated that an AD (P = 0.04) upon HNC diagnosis in early stage contributes to cumulated MME dose in the first year post-HNC diagnosis. This study underlines how anxiety has important repercussions on the management of pain and illustrates the importance of screening for AD upon HNC diagnosis to allow for early prophylactic treatment and support.


Method comparison of Particle Enhanced Immunoturbidimetry (PEIT) with High Performance Liquid Chromatography (HPLC) for glycated hemoglobin (HbA1c) analysis.
High Performance Liquid Chromatography (HPLC) technique is considered as a gold standard for HbA1c analysis however all laboratories cannot adopt it due to certain limitations. Our aim was to compare Particle Enhanced Immunoturbidimetry (PEIT) method with High Performance Liquid Chromatography (HPLC) for HbA1c analysis. All blood samples were analyzed by HPLC assay on a Bio-Rad D-10 analyzer and PEIT on an Erba XL-200 analyzer. Precision studies were undertaken and Coefficient of Variation (%CV) calculated. Systemic Error (SE), Random Error (RE) and Total Error (TEcalc) were obtained. The Total Allowable Error (TEa) set by the National Glycohemoglobin Standardization Program (NGSP) for HbA1c is 6%.The acceptable evaluation method is where TEcalc is less than TEa. RESULTS: The Precision studies were satisfactory with Coefficient of Variation (%CV) being less than 4% for both techniques. Mean HbA1c levels were slightly higher from HPLC than PEIT 9.07 ± 2.23% and 8.93 ± 2.10% respectively, although the difference was minimal. RE was 1.41%, TEcalc was 1.55%, which was less than TEa set by the NGSP. Both methods strongly correlated with the correlation coefficient (r) 0.9716, p < 0.0001. Our study showed HbA1c analysis by PEIT technique is precise, accurate, rapid and convenient and can be employed as an alternative to HPLC technique in countries where cost is a major problem for diagnosis and treatment.


The effect of noise interference, type of cognitive stressor, and order of task on cardiovascular activity.
In the search for reliable, easily applicable mental stressors many researchers have investigated the propensity of mental arithmetic and Stroop color discrimination tasks to elicit a physiological stress response; some researchers have added noise interference with the expectation of larger response magnitudes. The present study investigates a number of previously untested questions by directly comparing cardiovascular responses to mental arithmetic or the Stroop task with and without noise interference in a sample of 66 young adults. Half of all subjects were exposed to the additional noise interference, all subjects responded to both types of mental stressors. Blood pressure responses did not discriminate among the stressors but noise interference consistently augmented heart rate responses during mental arithmetic. Response to the second, repeated stressor was associated with highly significant response habituation effects for blood pressure. Furthermore, when mental arithmetic was presented first, both tasks elicited comparable and large responses; when the Stroop test was presented first both tasks triggered smaller responses but of comparable magnitude. Subjective evaluations of the stressors were not predictive of overall physiological response.


Operative transluminal balloon angioplasty. Adjunct to coronary bypass for extended myocardial revascularization of more than 3000 lesions in 1000 patients.
Operative transluminal coronary artery balloon angioplasty has been used for over 3000 lesions in 1000 patients since 1980. Initially it was only used for distal stenoses not accessible to coronary bypass grafting in 200 patients. Recatheterization of patients who had intraoperative transluminal balloon angioplasty of the proximal left anterior descending, right, and circumflex coronary arteries 3 years previously revealed excellent patency of both the bypass grafts and the dilated native coronary arteries. This observation supports the thesis that with properly constructed bypass anastomoses competitive flow does not significantly mandate graft thrombosis. Subsequently, intraoperative balloon angioplasty has been performed for both proximal and distal stenoses in 800 patients to improve native coronary artery perfusion and maximize revascularization. Follow-up from 1 to 7 years revealed perioperative myocardial infarction in 21 patients (2.1%) and death in 19 patients (1.9%). Recatheterization from 1 to 7 years after the operation in 51 patients (41 with symptoms) revealed that patency was almost as prevalent in arteries subjected to angioplasty (82%; 137/167) as in bypass grafts (84%; 102/122). Intraoperative balloon angioplasty appears to improve coronary artery perfusion without detrimental competitive flow when used with bypass grafts.


A two-stage Bayesian method for estimating accuracy and disease prevalence for two dependent dichotomous screening tests when the status of individuals who are negative on both tests is unverified.
Estimating the disease prevalence and test accuracy (sensitivity and specificity) for two dependent screening tests when the status of individuals who are negative on both tests is unverified represents a considerable challenge, as the disease rates for individuals negative on both tests are not identifiable without additional assumptions. This article presents a unified framework for handling this non-identifiability problem using two-step hierarchical informative prior on the sensitivities by two-stage Bayesian modeling with the characterized by joint testing strategies based on the inherent attribute of screening/diagnostic tests. We assign a diffuse and less risky two-step hierarchical informative uniform prior to the sensitivities while assigning a uniform (0,1) prior distribution to the specificities and prevalence. Strategies for model evaluation, general global evaluations, and individual cell checking are presented. Simulations are conducted under various scenarios to evaluate the performance of the proposed method. Applications to real data are also presented to illustrate the potential impact and benefit of the proposed method. Our results indicate that when the priors of sensitivities are assigned as appropriate two-step hierarchical informative priors, or even in the absence of the priors for the specificities and prevalence, the parameters involved in this study can still be estimated well. The advantages and limitations of this method in solving such problems are discussed and compared with other two-stage methods. We developed a two-stage Bayesian method for two dependent dichotomous screening tests with unverified individuals who are negative on both tests, and addressed the ad hoc model evaluation and checking procedures. The method can be understood easily and used conveniently by non-statisticians.


Patient-ventilator asynchrony in Saudi Arabia: Where we stand?
Patient-ventilator asynchrony in Saudi Arabia practices is common, and more emphasis on how to mitigate such a clinical problem is needed. This letter is intended to shed the light on the current national evidence of patient-ventilator asynchrony and how to step ahead for better patients' ventilation management.


'A plant's major strength in rhizosphere': the plant growth promoting rhizobacteria.
Human activities, industrialization and civilization have deteriorated the environment which eventually has led to alarming effects on plants and animals by heightened amounts of chemical pollutants and heavy metals in the environment, which create abiotic stress. Environmental conditions like drought, salinity, diminished macro-and micro-nutrients also contribute in abiotic stress, resulting in decrement of survival and growth of plants. Presence of pathogenic and competitive microorganisms, as well as pests lead to biotic stress and a plant alone can not defend itself. Thankfully, nature has rendered plant's rhizosphere with plant growth promoting rhizobacteria which maintain an allelopathic relationship with host plant to defend the plant and let it flourish in abiotic as well as biotic stress situations. This review discusses the mechanisms behind increase in plant growth via various direct and indirect traits expressed by associated microorganisms in the rhizosphere, along with their current scenario and promising future for sustainable agriculture. It also gives details of ten such bacterial species, viz. Acetobacter, Agrobacterium, Alcaligenes, Arthrobacter, Azospirillum, Azotobacter, Bacillus, Burkholderia, Enterobacter and Frankia, whose association with the host plants is famed for enhancing plant's growth and survival.


Usefulness of performing colonoscopy one year after endoscopic polypectomy.
This retrospective study defines a population with neoplastic colonic polyps who have had colonoscopic polypectomy and, in follow-up within one year, a repeat colonoscopic evaluation. The population was broken down into two groups, one group that had polyps at the second examination and one group that did not. This study determined which factor(s) were significant among this population in distinguishing whether new polyps would be found at one year follow-up. The authors found that among the many variables studied, only polyp multiplicity was significant in predicting polyp recurrence. More than one polyp found at index colonoscopy led to a significant chance of having a new polyp after only one year. Also, it was demonstrated that these "new" polyps were unlikely to have been "missed" polyps from the initial colonoscopy. Because of the shifting location, smaller size, and fewer instances of histologic atypia in these polyps compared with those at index examination, the authors believe that polyps found after one year may be assumed to have arisen de novo. Finally, the authors show that a significant number of polyps occur beyond the reach of the flexible sigmoidoscope (approximately 60 cm). The authors recommend that patients who have polyps undergo a colonoscopic examination. When patients are re-evaluated after having colonoscopic neoplastic polypectomy, they should undergo repeat colonoscopy.


Statins lower the risk of developing Alzheimer's disease by limiting lipid raft endocytosis and decreasing the neuronal spread of Herpes simplex virus type 1.
Many possible risk factors for Alzheimer's disease (AD) have been investigated, with only a very few showing positive associations and none defining the etiology of the neurodegenerative disease. The presence of herpes simplex virus type 1 (HSV-1) DNA in the brain, coupled with apolipoprotein E allele e4 (ApoE e4), has been suggested to confer an increased risk for AD. Studies have shown that pathogens, including viruses, utilize clathrin-independent endocytosis, i.e., lipid rafts that contain cholesterol, as part of their structure. Moreover, cholesterol-lowering statins have recently been linked with a reduced risk of developing Alzheimer's dementia. We, therefore, posit that long-term statin therapy protects individuals from AD by reducing the neuronal spread of HSV-1 via lipid raft domain pathways. Although the mechanism by which statins reduce AD risk is unknown, they reduce the amount of cholesterol in the plasma membrane and, thus, may decrease the availability of lipid raft pathways to spread HSV-1 within the brain.


Characterizing the differential distribution and targets of Sumo1 and Sumo2 in the mouse brain.
SUMOylation is an evolutionarily conserved eukaryotic posttranslational protein modification with broad biological relevance. Differentiating between the major small ubiquitin-like modifier (SUMO) paralogs and uncovering paralog-specific functions in vivo has long been very difficult. To overcome this problem, we generated His6-HA-Sumo2 and HA-Sumo2 knockin mouse lines, expanding upon our existing His6-HA-Sumo1 mouse line, to establish a "toolbox" for Sumo1-Sumo2 comparisons in vivo. Leveraging the specificity of the HA epitope, we performed whole-brain imaging and uncovered regional differences between Sumo1 and Sumo2 expression. At the subcellular level, Sumo2 was specifically detected in extranuclear compartments, including synapses. Immunoprecipitation coupled with mass spectrometry identified shared and specific neuronal targets of Sumo1 and Sumo2. Target validation using proximity ligation assays provided further insight into the subcellular distribution of neuronal Sumo2-conjugates. The mouse models and associated datasets provide a powerful framework to determine the native SUMO "code" in cells of the central nervous system.


Secondhand smoke exposure-induced nucleocytoplasmic shuttling of HMGB1 in a rat premature skin aging model.
Secondhand cigarette smoke exposure (SSE) has been linked to carcinogenic, oxidative, and inflammatory reactions. Herein, we investigated whether premature skin aging could be induced by SSE in a rat model, and assessed the cytoplasmic translocation of high mobility group box 1 (HMGB1) protein and collagen loss in skin tissues. Animals were divided into two groups: SSE and controls. Whole body SSE was carried out for 12 weeks. Dorsal skin tissue specimens were harvested for HMGB1 and Mallory's azan staining. Correlations between serum HMGB1 and collagen levels were determined. Rat skin exposed to secondhand smoke lost collagen bundles in the papillary dermis and collagen decreased significantly (p<0.05) compared with control rats. In epidermal keratinocytes, cytoplasmic HMGB1 staining was more diffuse and there were more HMGB1-positive cells after four weeks in SSE compared to control rats. A negative correlation between HMGB1 serum and collagen levels (r=-0.631, p=0.28) was also observed. Therefore, cytoplasmic HMGB1 expression in skin tissues might be associated with skin collagen loss upon the initiation of SSE. Additionally, long-term SSE might affect the appearance of the skin, or could accelerate the skin aging process.


Peptichemio in pretreated patients with plasmacell neoplasms.
Twenty-one patients with alkylator-resistant plasmacell neoplasms were treated with Peptichemio (PTC) at a dose of 40 mg/m2 for 3 days every 3 weeks or, in the case of persistent leukopenia and/or thrombocytopenia, at the single dose of 70 mg/m2 every 2-3 weeks according to haematological recovery. Seventeen patients, 10 with multiple myeloma and seven with extramedullary plasmacytoma (EMP), were fully evaluable. Six of 17 patients (35%) responded: three of seven EMP patients had a complete remission and 3 of 10 multiple myeloma patients had an objective response greater than 50%. The median duration of response was 8.5 months. An EMP patient obtained a complete response lasting for 16 months. The most frequent toxic effect were phlebosclerosis, occurring in all the patients, and myelosuppression, which was severe in only one case. PTC appears to be an active drug in patients with plasmacell neoplasms even if resistant to alkylating agents.


Trace elements exposure influences proximate body composition and antioxidant enzyme activities of the species tilapia and catfish in Burullus Lake-Egypt: human risk assessment for the consumers.
The trace elements concentration in the fish estimates the contamination degree in the aquatic environment. These toxic trace elements are transported into the human through the consumption of polluted fish. This study estimated the effect of Cd, Hg, Pb, As, and Al on tilapia species and catfish Clarias gariepinus (six for each species of fish) inhabiting Burullus Lake-Egypt 30° 33'-31° 08' E and 30° 22'-31° 35' N. The highest Pb concentrations recorded in the muscle of C. gariepinus 2.29 ± 0.29 μg/g while S. galilaeus was estimated the lowest Hg concentration of 0.54 ± 0.02 μg/g which indicated the presence of contaminants exceeded the limits permitted by FAO/WHO and EC. The maximum mean carbohydrate, lipid, and protein recorded in O. niloticus 18.66, 16.33, and 58.16 mg/g, respectively; moisture in O. aureus 67.33%; and ash 16.41% in O. niloticus. The lowest amount of carbohydrate was recorded in the T. zillii 14.1 mg/g, lipid, and ash in C. gariepinus 11.65 mg/g and 3.375%, respectively. Protein and moisture in the S. galilaeus were 53.75 mg/g and 60.75%, respectively. The results recorded a marked insignificant (P > 0.05) decrease in CAT, GR, and GPx activity in O. niloticus. GSH and SOD activity was an insignificant (P > 0.05) decrease in C. gariepinus. The results concluded that the trace elements concentrations exceed the maximum permissible limits recommended in fish samples set by Egypt, FAO, WHO, and EC. The estimated weekly intake of all elements through consumption of studied fish species inhabiting Burullus Lake by a child (15 kg) in Egypt is well above the PTWI recommended by FAO/WHO, whereas it is well below the PTWI for human consumption by young people (40 kg) and adult person (70 kg), at least in respect of residual levels of studied elements excluding Cd and Hg. Thus, for consumer protection, these fish species are unsafe and have hazardous effects for children, and about youth and adult consumption, caution must be taken to consider individuals eating significant amounts of fish.


Simple chemical synthesis and hybridization properties of non-radioactive DNA probes.
A simple chemical method for the synthesis of non-radioactive DNA probes is described: triazolyl-containing sequences were built by incorporation of 4-triazolylpyrimidin-2-ones instead of cytidines during oligodeoxyribonucleotide synthesis. The activating triazolyl groups were then displaced by a diamine which was further derivatized by a label, such as biotin. Synthesized DNA probes were oligonucleotides complementary to a cloned human antithrombin III DNA sequence. These probes, containing the same label at different positions of the sequence, were hybridized to their target DNA immobilized on nitrocellulose. Their hybridization specificity and stability were studied. Hybrid detection was performed either colorimetrically by the streptavidin-alkaline phosphatase-based system or by autoradiography after 5'-32P labeling of the probes: 15 fmol (0.05 microgram) of complementary sequence could be visualized in the two cases.


Botryoid-type of embryonal rhabdomyosarcoma of the proximal and mid ureter in an adult woman.
We report a unique case of botryoid-type embryonal RMS of the proximal and mid ureter in a pregnant 29-year-old woman. The ureteral polyp consisted of a malignant small blue round cell tumor with a myxoid background and contained evidence of foci of immature cartilage and aggregates of epithelial cells reminiscent of hair follicle. Immunohistochemical stains for myogenin and desmin confirmed skeletal muscle, or rhabdomyoblastic, differentiation. The compact epithelial cell fragments reminiscent of hair follicle differentiation were positive for p40. Treatment included 6 cycles of adjuvant chemotherapy (vincristine, actinomycin and cyclophosphamide (VAC). No recurrent or metastatic disease was identified post-surgery.


Human metallothionein gene MT1L mRNA is present in several human tissues but is unlikely to produce a metallothionein protein.
A human MT gene from the functional locus on chromosome 16, MT1L, is characterised and shown to produce mRNA in at least four human tissues. This gene is unlikely to produce a metallothionein protein because it contains a termination codon at position 26, by analogy to other human MT1 genes. MT1L cDNA is almost identical to another metallothionein cDNA clone reported recently, MT1R, suggesting that either there are unmapped human metallothionein genes, or that MT1L is polymorphic.


[Diffuse idiopathic skeletal hyperostosis].
Diffuse idiopathic skeletal hyperostosis is a progressive non-inflammatory bone and entheses disease which reduces the quality of life and self-sufficiency of patients. Despite significant prevalence is awareness of this disease still low and often leads to misdiagnosis. Therapeutic approach is different in diseases with similar symptomatology so it is important to raise awareness of this disease.


Effects of age on cell size and ion uptake in canine cortical bone.
In cortical bone from dogs of various ages, the uptakes of two bone-seeking isotopes, 85Sr and 99mTc-methylene diphosphonate (MDP), were compared with that of the intracellular cation 42K by using the volume of distribution (VD) technique. The rate of bone remodeling and the osteocyte volume were estimated by morphometric techniques. The percentage cell volume was related to the VD of 42K (r = 0.83; P less than 0.001) and decreased with increasing age. The decline in the VD of 85Sr and 99mTc-MDP with aging was related to the decrease in bone formation (r = 0.90; P less than 0.001). On a logarithmic plot, the cell volume and the VD of 42K had a linear relationship with the rate of new bone formation, the VD of 85Sr, and the VD of 99mTc-MDP. Our results demonstrate that, with increasing age in dogs, the bone formation rate and cell size decrease, as does the uptake of 42K, 99mTc-MDP, and 85Sr.


Elution of lipoprotein fractions containing apolipoproteins E and A-I in size exclusion on Superose 6 columns is sensitive to mobile phase pH and ionic strength.
Separation of lipoproteins secreted from McA-RH7777 (rat hepatoma) cells by Superose 6 column size-exclusion chromatography, using PBS buffer (NaCl 150 mM, sodium phosphate 10 mM, pH 7.5, EDTA 1 mM), produced apolipoprotein (apo) E or A-I profiles that did not correlate with lipoproteins separated by density ultracentrifugation. By density ultracentrifugation, apoE and apoA-I were mostly (> 90%) confined to high-density lipoproteins (HDL, d = 1.063-1.023 g/ml), but by chromatography apoE and apoA-I were recovered in all lipoprotein classes, including low-density lipoproteins (LDL), HDL, and post-HDL. Moreover, the elution volume of phenol red on Superose 6 greatly exceeded the total column volume. These discrepancies were attributable to pH and ionic strength effects. In low ionic strength, high pH buffer (Tris 25 mM, pH 8.3), elution volumes of lipoproteins, albumin, and phenol red were minimized. Elution volumes increased 25-70% when buffer pH was lowered at constant ionic strength (Tris 25 mM, pH 7.4) or when ionic strength was increased at constant pH (Tris 25 mM, pH 8.3, NaCl 500 mM). Altered phase partition appeared to cause the altered elution volumes, since recovery (measured as analyte peak area), resolution (measured as peak width at half height), and column void volume varied little from buffer to buffer. In Superose 6 size-exclusion chromatography with PBS buffer, then, elution volumes vary with pH and ionic strength. We propose that TBE buffer (Tris-borate 89 mM, pH 8.3, EDTA 2 mM) may produce fewer artefacts than PBS. With TBE there were (i) better correlation between size-exclusion and ultracentrifugal fractions, (ii) lower elution volumes, and (iii) less ¿smearing¿ of McA-RH7777 apoE and apoA-I containing lipoprotein bands.


Metabolic CO2 removal by dialysis: THAM vs NaOH infusion.
New methods of respiratory support are needed to reduce the high mortality rate of acute respiratory failure. To simplify the procedures of extracorporeal CO2 elimination under apneic oxygenation, one approach is to replace the membrane lung by a hemodialyzer and to administer an alkali, since hemodialysis requires a lower blood flow rate than blood-gas exchange. This study compared the effectiveness of trishydroxymethyl aminomethane (THAM) and NaOH in this procedure. Twelve male Anglo-Poitevin dogs (25 to 33 kg) were anesthetized, curarized and mechanically hypoventilated (VE = 41% of the control value). After not less than 15 min, a venovenous shunt was used for dialysis with blood flow of 7-10 ml. min.-1kg-1 for at least 8 hours. The dialysate contained no acetate, bicarbonate or lactate, but was alkalinized to a pH of 8-9 by the addition of NaOH. A solution of THAM (0.5 N) was infused into the right heart at the rate of 0.30 ml.min.-1kg-1 in six animals, and NaOH (0.15 N) was infused in the other six at the rate of 0.80 ml.min-1kg-1. The injected volumes were compensated for by an equivalent amount of ultrafiltration. Elimination of CO2 (mean TCO2 = 2.3 ml.min.-1kg-1) was the same with both methods and the difference for the electrolytes and acid-base equilibrium was only very small. However, hemolysis was six times greater with NaOH than with THAM. Despite ultrafiltration, a similar marked weight gain was observed from the second hour of the experiment in the NaOH series, but only after 7 hours with THAM. It thus appears that hemodialysis combined with alkalinisation is still too complex a procedure to be safely applied in acute or chronic pulmonary failure.


Preclinical Study on the Hepatoprotective Effect of Pollen Extract of Pinus brutia Ten. (Red Pine) in Mice and Phenolic Acid Analysis
Many agents, including those from herbal sources, have been sought as preventives or cures for hepatotoxicity. The pollen of Pinus brutia Ten., known as red pine (Pinaceae), is used against liver diseases in Anatolian folk medicine. In the current study, pollen ethanol extract of P. brutia was investigated for its possible hepatoprotective activity using a mouse model of CCl4-induced hepatotoxicity. Swiss albino mice were divided into five groups, and extract-treated groups were compared with a silymarin-treated group as the reference. The extract was tested at 100, 200, and 300 mg/kg (b.w.). Phenolic acids were analyzed using high-performance column chromatography (HPLC) in the extracts as pollens are usually known to be rich in phenolics. Our data revealed that the extract displayed the best hepatoprotection at a dose of 100 mg/kg when compared with silymarin (Legalon®), the reference drug. HPLC analysis indicated presence of protocatechuic acid (0.176 mg/g extract), p-hydroxybenzoic acid (0.001 mg/g extract), vanillic acid (VA) (0.537 mg/g extract), syringic acid (0.050 mg/g extract), and tr-cinnamic acid (0.310 mg/g extract), while the major phenolic acid was VA. The outcomes of this study allow us to conclude that red pine pollen extract can serve as a promising hepatoprotective agent. Among the phenolic acids analyzed in the pollen extract, vanillic acid as the major one besides some other phenolic acids detected seems to be responsible for its remarkable hepatoprotective effect.


Collection of platelets with a new cell separator and their storage in a citrate-plasticized container.
A new apheresis device using microprocessor control for the collection of a high-purity single-donor platelet concentrate was evaluated, as was the storage of platelets for up to 5 days in a citrate-plasticized polyvinylchloride blood bag. The study was conducted in three phases: collection of platelets for in vitro studies and determination of donor safety; autologous transfusion of platelets in healthy volunteers; and transfusion of platelets in patients requiring platelet transfusion therapy. Donors had mild hypocalcemia and minimal changes in blood counts except for a platelet count reduction from 288 +/- 50 x 10(3) (288 +/- 50 x 10(9)/L) to 217 +/- 43 x 10(3) per microL (217 +/- 43 x 10(9)/L). A mean of 3.36 +/- 1.24 x 10(11) platelets was collected in the mean volume of 214 mL with red cell and white cell contamination in the range of 10(7). Morphology and aggregation were as described previously in stored platelets. Platelet survival data in eight subjects showed a mean recovery of 61 +/- 11 percent and mean survival of 5.03 +/- 1.07 days by a weighted-mean model. Patients transfused with platelets had mean increments of 23,000 immediately and of 8000 at 24 hours; corrected count increments were 6000 at 1 hour and 4000 at 24 hours. The platelets were successful in providing hemostasis to these patients. Clinically useful 5-day-stored platelets are obtained by using this apheresis technology with a functionally closed system and a citrate-plasticized blood bag.


Virus-like particles in venom of Meteorus pulchricornis induce host hemocyte apoptosis.
Ultrastructural studies on the reproductive tract and venom apparatus of a female braconid, Meteorus pulchricornis, revealed that the parasitoid lacks the calyx region in its oviduct, but possesses a venom gland with two venom gland filaments and a venom reservoir filled with white and cloudy fluid. Its venom gland cell is concaved and has a lumen filled with numerous granules. Transmisson electron microscopic (TEM) observation revealed that virus-like particles (VLPs) were produced in venom gland cells. The virus-like particle observed in M. pulchricornis (MpVLP) is composed of membranous envelopes with two different parts: a high-density core and a whitish low-density part. The VLPs of M. pulchricornis is also found assembling ultimately in the lumen of venom gland cell. Microvilli were found thrusting into the lumen of the venom gland cell and seem to aid in driving the matured MpVLPs to the common duct of the venom gland filament. Injection of MpVLPs into non-parasitized Pseudaletia separata hosts induced apoptosis in hemocytes, particularly granulocytes (GRs). Rate of apoptosis induced in GRs peaked 48h after VLP injection. While a large part of the GR population collapsed due to apoptosis caused by MpVLPs, the plasmatocyte population was minimally affected. The capacity of MpVLPs to cause apoptosis in host's hemocytes was further demonstrated by a decrease ( approximately 10-fold) in ability of host hemocytes to encapsulate fluorescent latex beads when MpVLPs were present. Apparently, the reduced encapsulation ability was due to a decrease in the GR population resulting from MpVLP-induced apoptosis.


Imaging Manifestations of Acute and Chronic Renal Infection That Mimics Malignancy: How to Make the Diagnosis Using Computed Tomography and Magnetic Resonance Imaging.
To review the computed tomography and magnetic resonance imaging manifestations of acute and chronic renal infections that may mimic malignancy and to provide useful tips to establish an imaging diagnosis. Acute and chronic bacterial pyelonephritis are usually readily diagnosed clinically and on imaging when the diagnosis is suspected based upon clinical presentation. When unsuspected, focal, extensive or mass-like, acute and chronic bacterial pyelonephritis may mimic infiltrative tumours such as urothelial cell carcinoma (UCC), lymphoma, and metastatic disease. Infection may be suspected when patients are young and otherwise healthy when there is marked associated perinephric changes and in the absence of metastatic adenopathy or disease elsewhere in the abdomen and pelvis. Renal abscesses, from bacterial or atypical microbial agents, can appear as complex cystic renal masses mimicking cystic renal cell carcinoma. Associated inflammatory changes in and around the kidney and local invasion favour infection. Emphysematous pyelonephritis can mimic necrotic or fistulizing tumour; however, infection is more likely and should always be considered first. Xanthogranulomatous pyelonephritis can mimic malignancy when focal or multifocal and in cases without associated renal calculi. Malacoplakia is an inflammatory process that may mimic malignancy and should be considered in patients with chronic infection. Bacillus Calmette-Guerin (BCG)-induced pyelonephritis is rare but can mimic renal malignancy and should be considered in patients presenting with a renal mass when being treated with BCG for urinary bladder UCC.


An unusual case of random fire-setting behavior associated with lacunar stroke.
A case of a 47-year-old man with a sudden onset of a bizarre and random fire-setting behavior is reported. The man, who had been arrested on felony arson charges, complained of difficulties concentrating and of recent memory impairment. Axial T1-weighted magnetic resonance imaging showed a low intensity lacunar lesion in the genu and anterior limb of the left internal capsule. A neuropsychological test battery revealed lower than normal scores for executive functions, attention and memory, consistent with frontal lobe dysfunction. The recent onset of fire-setting behavior and the chronic nature of the lacunar lesion, together with an unremarkable performance on tests measuring executive functions two years prior, suggested a causal relationship between this organic brain lesion and the fire-setting behavior. The present case describes a rare and as yet unreported association between random impulse-driven fire-setting behavior and damage to the left internal capsule and suggests a disconnection of frontal lobe structures as a possible pathogenic mechanism.


Spatial dynamics of multistage cell lineages in tissue stratification.
In developing and self-renewing tissues, terminally differentiated (TD) cell types are typically specified through the actions of multistage cell lineages. Such lineages commonly include a stem cell and multiple progenitor (transit-amplifying) cell stages, which ultimately give rise to TD cells. As the tissue reaches a tightly controlled steady-state size, cells at different lineage stages assume distinct spatial locations within the tissue. Although tissue stratification appears to be genetically specified, the underlying mechanisms that direct tissue lamination are not yet completely understood. Herein, we use modeling and simulations to explore several potential mechanisms that can be utilized to create stratification during developmental or regenerative growth in general systems and in the model system, the olfactory epithelium of mouse. Our results show that tissue stratification can be generated and maintained through controlling spatial distribution of diffusive signaling molecules that regulate the proliferation of each cell type within the lineage. The ability of feedback molecules to stratify a tissue is dependent on a low TD death rate: high death rates decrease tissue lamination. Regulation of the cell cycle lengths of stem cells by feedback signals can lead to transient accumulation of stem cells near the base and apex of tissue.


Exposure to Landfill Leachates Affects the Embryonic Development of Zebrafish, Danio rerio: A Case Study in Yucatan, Mexico.
We report the chemical characterisation and toxic effects of municipal solid waste landfill leachates on the embryonic development of Danio rerio. The results of the Fourier transform infrared spectroscopy (FTIR) revealed the presence of nitrogen-containing groups and aromatic functional groups associated with highly toxic pollutants such as ammonia and heavy metal-humic complexes. Mortalities of up to 93 and 100% were observed in 1:64 and 1:32 landfill leachate dilutions, v/v, respectively. The hatching percentages of the fish were also severely affected, with very low percentages ranging from 0 to 33.3% for 1:32, 1:64 and 1:128 dilutions, v/v. Morphologically, a developmental arrest was evident for all treatments. This study reveals the high toxicity of landfill leachates that could contaminate the aquifer of the Yucatan Peninsula and threaten the health of living organisms.


A Turner syndrome woman with a ring X chromosome [45,X/46,X,r(X)(p22.3q27)] whose child also had a ring X chromosome.
To describe a woman with Turner syndrome with ring X chromosome mosaicism who had a child who possessed the same ring X chromosome. Polymorphisms of genes located on the X chromosome from genomic DNA of the mother, father, and the child were evaluated. The mother's karyotype was 45,X [48]/46,X,r(X)(p22.3q27) [2], and the child's karyotype was 45,X[33]/46,X,r(X)(p22.3q27) [17]. Polymerase chain reaction was used to amplify short tandem repeats from the loci of the hypoxanthine phosphoribosyltransferase gene and the androgen receptor gene. Alleles for both genes in the child originated from both parents in a heterozygous fashion. The alleles originating from the mother originated from the ring X chromosome. However, the amount of amplified DNA was less than that of a normal X chromosome. The ring X chromosome of the mother was most likely transmitted to the newborn. Thus, an ovum with the ring X chromosome can be fertile and can produce a viable zygote.


Lead-contaminated soils with contrasting texture remediated with phosphate: chemical fractionation and chloropyromorphite stability.
Pb can be stabilized in soil as Pb-P mineral. The aims of this study were to access the distribution of Pb in organic and mineral fractions of contrasting texture of soil Pb-contaminated and remediated with P and Cl and to evaluate the stability of chloropyromorphite in these soils. A clay loam Oxisol (sandstone) and a clayey Ultisol (basalt) were used in a factorial experiment, with three replications: two soils, two Pb contamination levels, two soil pH values, and four P doses. The Pb concentrations were determined in seven soil phases. Release kinetics of Pb were performed with 0.1 mol L-1 pH 2.5 citric acid. The transfer of soil Pb to chloropyromorphite was dependent on the level of contamination in the clay loam Oxisol. In the lowest P dose (molar ratios P:Pb 3:5), the main source was the Pb complexed in the organic matter and in the highest P dose (molar ratios P:Pb 12:5) was the Pb adsorbed by inner sphere in gibbsite and kaolinite. The release of Pb in the citric acid was dependent on the texture and mineralogy of the soils. Pb recovery applied to the clay loam Oxisol was around 100% (biphasic kinetic), while for the clayey Ultisol, the recovery ranged from 43 to 52% (single-phase kinetic). Remediation of Pb-contaminated soils with P and Cl is more efficient in clayey and oxidic soils since chloropyromorphite formation is faster and its solubilization is slower, an important combination in environmental terms.


Economic impact of faith-based residential addiction recovery for the homeless.
To examine the economic and fiscal impact of a faith-based long-term residential addiction recovery program for homeless men on the state and local economy. This descriptive study used entry, within-program, and exit data to determine the program's fiscal and economic impact. The population consisted of 5,122 homeless men at an inner-city mission in Baltimore City from 2006 to 2019. To compare costs and savings at different program intervals, economists examined program data before, during, and following completion of the program. Data were analyzed using the IMPLAN input-output model. The study revealed that each man who participates in the residential recovery program for 1 year produces a total savings of $14,263 to state and county governments. Each man who successfully completes the program saves state and county governments over $5,000 per year post graduation. Each man who works for 1 year following graduation and reallocates his spending from illicit drugs to goods and services in the economy supports 1.5 jobs and generates over $240,000 in additional revenue and economic output. The findings from this study support the value of this faith-based residential program in promoting economic productivity in the surrounding state and region.


Effects of hypoxia on cardiac vagal efferent activity and on the action of the vagus nerve at the heart in the dog.
The discharge of cardiac efferent fibres in the central end of the transected vagus nerve was studied in single or few-fibre preparations in anaesthetized dogs. At the same time standard electrical stimuli were delivered to the cardiac end of the vagus nerve. During progressive asphyxic hypoxia induced by tracheal occlusion cardiac vagal activity increased and vagal action on the heart was potentiated.


Investigation of iron uptake and virulence gene factors (fur, tonB, exbD, exbB, hgbA, hgbB1, hgbB2 and tbpA) among isolates of Pasteurella multocida from Iran.
Iron is an essential compound in metabolic pathway of wide range of organisms. Because of limited free iron supply in mammalian and avian hosts, bacteria have applied various ways to acquire iron. In this study, the frequency of 8 iron acquisition factors was examined among 63 avian and ovine Pasteurella multocida field isolates and their vaccine strains using PCR method. Five candidate genes (fur, tonB, exbD, exbB and hgbA) were identified among all isolates. For the first time, 2 loci (hgbB1 and hgbB2) of the hgbB gene were identified, which were previously reported as 1 gene. Also, it was found that 5 ovine and 1 avian isolates possessed all the virulence factors, which could also be considered for evaluating the frequency of other virulence factors. More studies need to be conducted on the frequency of all other virulence factors among these isolates, which can provide basic information for improvement or substitution of current vaccinal strains. Overall, as the new designed sets of primers showed more potential in detecting the corresponded genes, researchers can consider them in further studies.


Evaluation of a new complement-dependent lymphocytotoxicity cross match method using an automated cell counter, the NucleoCounter® NC-3000™.
Complement-dependent lymphocytotoxicity cross match (CDC-XM) is the ultimate test of donor/recipient compatibility prior to organ transplantation. This test is based on cell viability, evaluated under fluorescence microscopy by an operator after proper staining. The determination of the positivity threshold may vary depending on the operator. We developed a new method in which the final step of determining cell viability is automated using the NC-3000™ (Chemometec®), an image cytometer able to precisely determine the percentage of dead/live cells in a suspension. After T and B donor cells isolation by negative selection, complement-dependent lysis was performed in macrovolumes in a PCR plate. Then, cell viability was measured by the NC-3000™. The sensitivity and routine CDC-XM results of this new method were compared to those of CDC-XM reference method using Terasaki plates. The sensitivity of CDC-XM expressed in the ASHI scoring system of this method was similar to the reference method results for a dilution range of the positive controls. Similarly, the results of the new method were comparable in a clinical situation to those obtained with the reference method after a study of 10 cross-matches, of which 5 cross-matches with DSA were positive and five cross-matches without DSA were negative. Moreover, ASHI scores were similar to those obtained using the reference method, and the mortality percentage was reproducible (CV < 15%). The assessment of cell viability by the NC-3000™ is easy to perform and highly reproducible but requires CDC-XM to be performed by the macrovolume method. The determination of a precise percentage of viability/mortality by the automation excludes operator variability and allows a better understanding of results close to the decision threshold.


Complete Genome Sequence of Escherichia coli Myophage Mansfield.
Mansfield is a PB1-like Escherichia bacteriophage with a 68,120-bp genome and a predicted 3,673-bp direct terminal repeat. This myophage encodes 105 proteins, for which 32 functions were predicted.


Aggressive infantile fibromatosis of the submandibular region.
Aggressive infantile fibromatosis is an unusual, tumor-like, proliferative lesion which grows infiltratively. The clinical and histological features of aggressive infantile fibromatosis in the submandibular region of a 3-year-old girl are reported.


[Cortical tissue from syngenous brain as a stimulator of immunogenesis in thymectomized mice].
Experiments on CBA mice showed that the gray matter homogenate of the syngeneic brain contained components reconstituting the T-cell population in the spleen of thymectomized animals and possessing a stimulating effect on the immune response to sheep red blood cells. The white matter homogenate was much less active, it is not excluded that the trace activity is due to the gray matter admixture in the preparation. Syngeneic muscle tissue homogenate possessed no biological activity.


Enhanced neogenic potential of Panc-1 cells supplemented with human umbilical cord blood serum--An alternative to FCS.
The promise(s) of using Fetal Calf Serum (FCS) as a supplement for the maintenance of cell cultures has been well documented. However, FCS forms the xenogenic source for any human derived cells/organ and limits its application. Recently, the usage of human umbilical cord blood serum (hUCBS) for maintenance of mesenchymal cells has been supportive. In the present study we investigated the effects of hUCBS and FCS on the proliferation (viability, proliferative) and its differentiation potential (DTZ staining, immunofluroscence) to generate islet like cellular aggregates (ICAs) using the human derived Panc-1 cell lines. A comparative analysis of hUCBS and FCS for each parameter demonstrated that hUCBS supplemented media was better for proliferation and differentiation of the Panc-1 cells. The ICAs obtained from hUCBS primed cultures showed a higher yield, increased islet size, and showed an increase for insulin staining compared to FCS. We suggest that hUCBS can be explored as an alternate serum supplement for FCS, making it more feasible in cell systems of human derived origin and can also find its application for the human transplantation programmes.