Erratum regarding missing Declaration of Competing Interest statements in previously published articles.
[This corrects the article DOI: 10.1016/j.xnsj.2020.100030.][This corrects the article DOI: 10.1016/j.xnsj.2020.100006.][This corrects the article DOI: 10.1016/j.xnsj.2020.100026.].


Lab-on-a-chip: applications in proteomics.
Recent advances in chip-based separation of proteins provide methods that are faster and more convenient than conventional gel electrophoresis. Rapid and automated protein sizing on a chip is at the commercial stage and first attempts have been made to perform two-dimensional separation on a chip. Numerous designs have been described to interface a microfluidic chip to a mass spectrometer. Impressive integration efforts are demonstrated by the ability to perform on-chip trypsin digestion, separation and injection into a mass spectrometer with a single device.


Engineering the Surface Properties of DNA Nanostructures by Tuning the Valency of Assembling Species for Biomedical Applications.
Self-assembled DNA nanostructures hold great potentials in biomedical applications. Nevertheless, the negatively charged DNA backbone and susceptivity to enzyme degradation pose challenges to this regard. Engineering the surface properties of DNA nanostructures by assembling DNA with guest molecules in magnesium-free system is promising to solve these issues. In this study, the polyamines-mediated DNA self-assembly with an emphasis on the valency of polyamines is investigated. Both spermine, spermidine, and putrescine can assemble DNA tetrahedron under appropriate concentrations. The cytotoxicity and cellular uptake efficiencies vary with the polyamine valency. Compared with magnesium-assembled DNA tetrahedron, polyamine-assembled DNA tetrahedron exhibits higher cellular uptake efficiency and serum stability. Circular dichroism spectrum results indicate that polyamines induce DNA conformation slightly shifting from B form to A form. The improved performances of polyamine-assembled DNA tetrahedrons under physiological settings are attributed to the surface properties that altered by guest molecules polyamine. The current study suggests that engineering the surface properties of DNA nanostructures by assembling them with guest cationic species is promising to further their biomedical applications.


Comparison of base inclination of ribo-AU and deoxyribo-AT polymers.
The inclination angle between the base normal and the helix axis is measured for ribo-AU polymers by using flow linear dichroism (LD), and compared to measurements for deoxyribo-AT polymers under dehydrating conditions. The CD of the DNA polymers under the dehydrating conditions is not the same as the corresponding RNA polymers, which are presumed to be in the A form. However, the LD indicates that poly(dAdT)-poly(dAdT) can assume the A form in 80% 2,2,2-trifluoroethanol, although poly(dA)-poly(dT) retains B form structure in this dehydrating solvent. The inclination angles are similar for B form poly(dAdT)-poly(dAdT) and poly(dA)-poly(dT), and these parameters are also similar for A form poly(rArU)-poly(rArU) and poly(rA)-poly(rU). All inclination axes are similar.


Lead acetate may cause erectile dysfunction by modulating NO/cGMP pathway in rat corpus cavernosum.
Despite the fact that metal toxicity has been widely reported in industrial toxicological studies, very little has been reported about the effect of lead exposure on erectile function. This study investigated the effect of lead on erectile function in rats and aimed to preliminarily test the mechanisms by which it might affect erection. Rats were injected with lead acetate (0.25-2 mg/kg) intraperitoneally for 21 days. Intracavernosal pressure/mean arterial pressure (ICP/MAP) next to nerve stimulation; nitrite/nitrate; malonaldehyde; and reduced glutathione levels and superoxide dismutase activity in the corpus cavernosum, kidney, and brain were measured in addition to creatinine, urea, and testosterone. For acute studies, rats were injected intravenously with lead acetate, and then ICP/MAP was recorded for 45 min. Subacute treatment significantly reduced erection with significant elevation of malonaldehyde and reduction of nitrite/nitrate levels in the corpus cavernosum. In acute studies, lead (2 and 5 mg/kg) reduced neurogenic erections by 28.42 ± 3.76 and 96.84 ± 8.52%, respectively, an effect that was masked in the presence of NG-nitro-L-arginine, tetraethyl ammonium, or methylene blue, but not zinc protoporphyrine, and reversed by vitamin C and partially by sildenafil. Lead acetate may inhibit the erectile process in rats. Besides its prooxidant effect and consequent inactivation of nitric oxide, lead may negatively modulate the action of nitric oxide on guanylate cyclase and potassium channels.


Distribution characteristics of methamphetamine and amphetamine in urine and hair specimens collected from alleged methamphetamine users in northern Taiwan.
This study was conducted to better understand the distribution characteristics of methamphetamine and amphetamine in urine and hair specimens collected from alleged methamphetamine users in the local population. It is anticipated that the data hereby obtained will be helpful to the interpretation of the time and pattern of drug use. Eight alleged methamphetamine-using arrestees from Keelung Police Department (north of Taipei, Taiwan) consented to contribute both urine and hair specimens. Each arrestee contributed seven urine specimens collected at 0, 12, 24, 48, 72, 96, and 120 h, respectively, after the arrest. Hair specimens were cut into 2-cm sections. The limits of detection and quantitation of the urine protocol were 40 and 50 ng/mL, respectively, for both amphetamine and methamphetamine, while the corresponding limits of detection and quantitation for the hair protocol were 0.8 and 1.0 ng/mg, respectively. The concentration variations of methamphetamine and amphetamine in the urine specimens exhibited three distinct patterns: (a) continuous decrease in the analytes' concentrations for specimens collected at hours 0-120; (b) increase in the analytes' concentrations in specimens collected at hours 0-12, followed by decrease; (c) increase in analytes' concentrations in specimens collected at later times. Together with the amphetamine/methamphetamine concentration ratios found in these urine specimens, the observed trends in the changes of the analytes' concentrations are helpful for the interpretation on the time of drug use. Unlike urine specimens, amphetamine/methamphetamine concentration ratios in various hair specimens and hair sections remain relatively constant.


A novel Doppler echocardiographic index integrating left and right ventricular function is superior to conventional indices for predicting adverse outcome of acute myocardial infarction.
The ratio of peak diastolic early velocity (E) of left ventricular (LV) inflow to peak diastolic longitudinal velocity (e') of the mitral annulus (E/e') is thought to reflect LV filling pressure, and tricuspid annulus velocity at systole (s'-T) is thought to reflect right ventricular function. However, it has not been reported on significance of the combined use of E/e' and (s'-T) to predict outcome of acute myocardial infarction (AMI). Over 21 months, beginning in January 2007, we enrolled 65 AMI patients who were measured hemodynamic and echocardiographic parameters by Swan-Ganz (SG) catheterization just after reperfusion therapy and echocardiography immediately after reperfusion therapy. Cardiac index (CI) and pulmonary capillary wedge pressure (PCWP) via SG catheter were measured, and routine echocardiographic indices, including E, e', E/e', and (s'-T) were determined. In addition, we defined the functional integrated bi-myocardial tissue Doppler (FIT) index as (s'-T)÷E/e'. The relationships between CI, PCWP, and echocardiographic indices were investigated, including FIT index. Moreover, we investigated whether FIT index could predict adverse cardiac events. FIT index was significantly associated with not only CI but also PCWP. In the Cox proportional hazards model, FIT index<1.0 was a significant predictor for adverse outcome of AMI after adjustment for age, Killip class, history of previous coronary revascularization, location of culprit lesion, and LV ejection fraction. The novel index defined as (s'-T)÷E/e' could be quite useful predictor of prognosis in AMI.


A sequential study of changes in the brain and cerebrospinal fluid of the rat following triethyltin poisoning.
Following a single intravenous (i.v.) injection of triethyltin (10 mg/kg) in rats, vacuoles appeared in the myelin sheath within 3 h and they progressively increased in size between 6 and 24 h. Their development was closely correlated with a progressive increase in water, sodium, and chloride content of the brain, and of cerebrospinal fluid (CSF) pressure. Oedema was more extensive in tissues consisting predominantly of white matter rather than grey matter. The sustained increase in CSF pressure did not precede the development of lesions and was the result of, rather than the cause of, brain oedema and swelling. These findings indicate that triethyltin has a direct effect on the myelin sheath.


Successful treatment of AIDS-related cryptosporidial sclerosing cholangitis.
Cryptosporidium is a well recognized cause of AIDS-related sclerosing cholangitis for which no efficacious therapy exists at present. We evaluated the efficiency of the combination of paromomycin and letrazuril in the treatment of this condition. We report a case of cryptosporidial cholangitis in a patient with AIDS who responded to treatment with intravenous paromomycin followed by oral letrazuril as maintenance therapy. The combination of paromomycin and letrazuril should be considered in the treatment of AIDS-related sclerosing cholangitis.


Changing epidemiology of parenterally transmitted viral hepatitis: results from the hepatitis surveillance system in Italy.
In 1991, compulsory hepatitis B virus vaccination and screening for anti-hepatitis C virus of blood banks were introduced in Italy. To evaluate the impact of preventive measures on the incidence and risk factors for parenterally transmitted viral hepatitis. Data from the surveillance system for acute viral hepatitis for the period 1985-99 were used. Temporal trends in distribution of reported risk factors were analysed by comparing three-year periods: 1987-89 and 1997-99. The incidence (no. cases per 100,000 population) of hepatitis B was 12 in 1985 and 3 in 1999; the incidence of hepatitis non-A, non-B decreased from 5 to 1 in the same period. These decreases were more evident among young adults and before rather than after 1991. Multiple sexual partners, other parenteral exposures and dental treatment remain the most common risk factors for parenterally transmitted viral hepatitis. An increase in frequency over time was observed for other parenteral exposures, whereas a marked decrease was evident for blood transfusion and household contact with an HB-sAg carrier. Invasive medical procedures continue to represent an important source of infection. Intravenous drug use was reported particularly by young adults with non-A, non-B hepatitis, with increased frequency over time. Non-immunologic measures for preventing hepatitis B and non-A, non B due to iatrogenic and other parenteral exposures, combined with hepatitis B virus vaccination, could further reduce parenteral transmission.


The Use of Bowel in Urologic Reconstructive Surgery.
Intestinal surgery involves an operative space shared by both general surgeons and urologists and is a border region where these 2 surgical disciplines often intersect. Urologists routinely use both small and large bowel for reconstructive procedures and surgeons often encounter such reconstructions of the urinary tract. It is essential for surgeons to understand the urologic indications for using intestinal segments for reconstructive procedures, the variety of such reconstructions, the anatomic landmarks and potential pitfalls that should be considered when intraoperatively encountering such reconstructions, and the potential metabolic consequences of the incorporation of bowel segments into the urinary collecting system.


An improved in situ measurement of offset phase shift towards quantitative damping-measurement with AFM.
An improved approach is introduced in damping measurement with atomic force microscope (AFM) for the in situ measurement of the offset phase shift needed for determining the intrinsic mechanical damping in nanoscale materials. The offset phase shift is defined and measured at a point of zero contact force according to the deflection part of the AFM force plot. It is shown that such defined offset phase shift is independent of the type of sample material, varied from hard to relatively soft materials in this study. This improved approach allows the self-calibrated and quantitative damping measurement with AFM. The ability of dynamic mechanical analysis for the measurement of damping in isolated one-dimensional nanostructures, e.g. individual multiwalled carbon nanotubes, was demonstrated.


Plaque-reduction assays for human and simian immunodeficiency virus neutralization.
Research on HIV vaccines, as well as studies on HIV pathogenesis in human and SIV in the macaque model, require the availability of simple and standardized assays for quantification of neutralizing antibodies to primary virus isolates. We have recently developed and standardized assays using human cell lines engineered to express CD4 and co-receptors for HIV and SIV entry. One cell line originated from a glioma (U87) and the other from an osteosarcoma (HOS). Both cell lines and their derivatives form monolayer cultures, a prerequisite for counting plaques. HIV-infected U87.CD4-CCR5 or -CXCR4 cells form syncytia, that is, plaques that can be stained with hematoxylin and enumerated by light microscopy. In addition to CD4 and co-receptors (most often used CCR5 and CXCR6 by SIV), GHOST(3) cells have been engineered to express the green fluorescent protein following virus infection. Infected cells show green fluorescence and can be enumerated by fluorescence microscopy. Neutralization is determined by the ability of a serum to reduce the number of plaque-forming units (PFU) relative to controls exposed to medium or negative serum. Both assays are run in microtiter format and neutralization is evaluated after 3 d. Intra-assay variation has been used for estimation of the cutoff for neutralization. Testing 15 serum-virus combinations in the U87.CD4 assay and four serum-virus combinations in the GHOST(3) assay revealed that standard deviation of differences ranged from 9.1% to 9.9% in the two assays. This allowed the use of a cutoff >3 SD; that is, 30% neutralization. Virus titration experiments showed that neutralization results were dependent on virus dose and therefore the neutralization assays should be performed with a virus dose of 10-100 PFU/well. The assays have high specificity and reproducibility, and are simple and sensitive high-throughput assays.


Adenovirus-mediated expression of keratinocyte growth factor promotes secondary flap necrotic wound healing in an extended animal model.
No effective treatments have been found for flap necrosis. Animal models that focus on the initial flap viability are inappropriate for necrotic wound studies. Keratinocyte growth factor (KGF) promotes keratinocyte proliferation with stronger activity and fewer complications and thus may be useful for necrotic flap wound healing. Rats with modified flap necrosis were randomly divided into four groups. An adenoviral vector expressing KGF was injected subdermally in the back of the animals after necrosis began. The expression and effect of KGF was assessed by real-time polymerase chain reaction, enzyme-linked immunoassay, and transwell, and wound healing was monitored. The plasmid and adenovirus were able to express KGF and stimulate epithelial cell growth (p = 0.029). Histology showed that the necrosis healed fastest in the KGF administration group than in the control groups (p < 0.01). The adenovirus-mediated KGF (Ad-KGF) group had the thickest epithelium on days 15 (p = 0.044) and 25 (p = 0.014). The KGF level in the blood serum soared 10 and 15 days postoperatively (p < 0.01) but returned to baseline by day 25 (p = 0.561). The KGF mRNA levels in vivo increased dramatically in the Ad-KGF group (p = 0.037). The extended flap model is applicable in necrotic wound study. Keratinocyte growth factor can promote secondary necrotic flap wound healing, and administration of KGF can be achieved by an adenoviral vector.


Treatment results for 84 patients with base of tongue cancer.
For T2 tumors, surgery was indicated if functional preservation was possible. For T3/T4 tumors, the rate of primary disease control was not high and surgery frequently involved total laryngectomy. Points that surgeons must heed when performing such surgery were delineated. Because tumors originating from the base of the tongue are rare, few large-scale studies of such tumors have been performed. We reviewed treatments and outcomes at our department to establish effective future therapeutic plans. From 1971 to 2000, 84 patients with previously untreated and resectable squamous cell carcinoma of the base of the tongue were treated at the Head and Neck Department of the Cancer Institute Hospital, Tokyo. Treatment selection and results were investigated. The main treatment options were radiotherapy for primary lesions < or = T2 and surgery for primary lesions > or = T3. Overall disease-specific 5-year cumulative survival rate was 59.8%, but there was no significant difference in survival rate at each stage between the two treatments. Among patients who died of the primary disease, the area that was most difficult to control was the superior margin of the lateral wall of the oropharynx (n=7). The incidence of contralateral or retropharyngeal lymph node metastasis was low if tumors neither crossed the midline nor infiltrated the lateral wall. While total laryngectomy was performed on 48 patients, the larynx was operatively preserved in 5 T3 patients and one T4 patient.


Correction: TROP2 promotes proliferation, migration and metastasis of gallbladder cancer cells by regulating PI3K/AKT pathway and inducing EMT.
[This corrects the article DOI: 10.18632/oncotarget.16789.].


High-performance liquid chromatographic determination of glyoxylic acid and other carbonyl compounds in urine.
A high-performance liquid chromatographic method for the determination of carbonyl compounds, namely aldehydes, ketones and keto acids in urine, has been developed. Pre-column derivatization with 2,4-dinitrophenylhydrazine offers sufficient sensitivity for the determination of glyoxylic acid in urine with a detection limit of 0.5 mg/l. The separation is performed on a C18 10-micron column and with an acetonitrile-aqueous buffer mobile phase, which also allows the resolution of the syn and anti geometric isomers. Matrix effects, precision, accuracy, limits of detection and structural selectivity of the method are discussed.


Why dietary restriction substantially increases longevity in animal models but won't in humans.
Caloric restriction (CR) extends maximum longevity and slows aging in mice, rats, and numerous non-mammalian taxa. The apparent generality of the longevity-increasing effects of CR has prompted speculation that similar results could be obtained in humans. Longevity, however, is not a trait that exists in a vacuum; it evolves as part of a life history and the physiological mechanisms that determine longevity are undoubtedly complex. Longevity is intertwined with reproduction and there is a cost to reproduction. The impact of this cost on longevity can be age-independent or age-dependent. Given the complexity of the physiology underlying reproductive costs and other mechanisms affecting life history, it is difficult to construct a simple model for the relationship between the particulars of the physiology involved and patterns of mortality. Consequently, we develop a hypothesis-neutral model describing the relationship between diet and longevity. Applying this general model to the special case of human longevity and diet indicates that the benefits of caloric restriction in humans would be quantitatively small.


Hydration status assessment by multi-frequency bioimpedance in patients with advanced chronic kidney disease.
Body composition assessment has the potential to improve the care of patients with chronic kidney disease (CKD). Whole-body multiple-frequency bioimpedance spectroscopy (BIS) appears to be a useful and appropriate technique for assessing hydration status and body composition in CKD patients. The aims of this study were to determine the hydration status by BIS in patients with advanced CKD, and to analyse the association of body fluid status with common clinical and biochemical characteristics. The prognostic value of the phase angle at 50 KHz (PA) was also evaluated. The study group consisted of 175 patients (66 ± 14 year, 77 females) with eGFR < 40 ml/min not yet on dialysis. Body composition was assessed by BIS (BCM, Fresenius). Hydration status was expressed as a percentage of the total body water (TBW). Patients were prospectively followed-up for a median of 481 days, and the main determinants of mortality were estimated by Cox regression analysis. The majority of patients (85%) showed a hydration status within ± 5% TBW. Patients with oedemas or uncontrolled arterial hypertension showed mean estimate fluid overload significantly higher than that of the other study patients. Fluid overload was negatively associated with serum albumin levels, body mass index and urinary sodium/potassium ratio; and positively with male gender and diabetes. During the follow-up period, 16 patients died (9%). The main determinants of mortality adjusted for other potential covariates were: Davies comorbidity index (HR = 4.304; P = .001), and PA (per each °; HR = 0.491; P = .026). BIS may help identify changes in hydration status in CKD patients not fully appreciated by clinical or biochemical assessment. PA was a significant predictor of mortality in these patients.


Forest floor decomposition, metal exchangeability, and metal bioaccumulation by exotic earthworms: Amynthas agrestis and Lumbricus rubellus.
Earthworms have the potential to reduce the retention of pollutant and plant essential metals in the forest floor (organic horizons) by decomposing organic matter and increasing exchangeability of metals. We conducted a laboratory experiment to investigate the effects of two exotic earthworms, Amynthas agrestis and Lumbricus rubellus, on forest floor decomposition, metal exchangeability, and metal bioaccumulation. Eighty-one pots containing homogenized forest floor material were incubated for 20, 40, or 80 days under three treatments: no earthworms, A. agrestis added, or L. rubellus added. For earthworm treatments, A. agrestis and L. rubellus were stocked at densities observed in previous field studies. Pots containing either A. agrestis or L. rubellus had lost more forest floor mass than the control plots after 40 and 80 days of incubation. Forest floor pots containing A. agrestis had significantly lower % C (16 ± 1.5 %) than control pots (21 ± 1.2 %) after 80 days. However, L. rubellus consumed more forest floor and C mass than A. agrestis, when evaluated on a per earthworm biomass basis. Exchangeable (0.1 M KCl + 0.01 M AcOH extractable) and stable (15 M HNO3+ 10 M HCl extractable) concentrations of Al, Ca, Cd, Cu, Mg, Mn, Pb, and Zn in forest floor material were measured. Stable concentrations and % exchangeable metals in forest floor material were similar among treatments. Although exchangeable metal concentrations varied significantly for most metals among treatments (except Mg and Zn), we conclude that earthworms did not increase or decrease the exchangeability of metals. However, earthworms bioaccumulated Cu, Cd, Zn, and Mg and had potentially hazardous tissue concentrations of Al and Pb. This was best illustrated by calculating bioaccumulation factors using exchangeable concentrations rather than total concentrations. Future research is needed to understand the effect of earthworms on metals in other soil types.


Interaction between delta-6-tetrahydrocannabinol (delta-6-THC) and lithium at the blood brain barrier in rats.
Neither the acute nor the chronic i.p. administration of delta-6-tetrahydrocannabinol affected the passage of lithium from blood to brain in normal rats.


Effects of hypoxia and reoxygenation on mitochondrial functions and transcriptional profiles of isolated brain and muscle porcine cells.
Oxygen fluctuations might occur in mammalian tissues under physiological (e.g. at high altitudes) or pathological (e.g. ischemia-reperfusion) conditions. Mitochondria are the key target and potential amplifiers of hypoxia-reoxygenation (H-R) stress. Understanding the mitochondrial responses to H-R stress is important for identifying adaptive mechanisms and potential therapeutic solutions for pathologies associated with oxygen fluctuations. We explored metabolic response to H-R stress in two tissue types (muscle and brain) with different degrees of hypoxia tolerance in a domestic pig Sus scrofa focusing on the cellular responses independent of the systemic regulatory mechanisms. Isolated cells from the skeletal muscle (masseter) and brain (thalamus) were exposed to acute short-term (15 min) hypoxia followed by reoxygenation. The mitochondrial oxygen consumption, reactive oxygen species (ROS) production rates and transcriptional profiles of hypoxia-responsive mRNA and miRNA were determined. Mitochondria of the porcine brain cells showed a decrease in the resting respiration and ATP synthesis capacity whereas the mitochondria from the muscle cells showed robust respiration and less susceptibility to H-R stress. ROS production was not affected by the short-term H-R stress in the brain or muscle cells. Transcriptionally, prolyl hydroxylase domain protein EGLN3 was upregulated during hypoxia and suppressed during reoxygenation in porcine muscle cells. The decline in EGLN3 mRNA during reoxygenation was accompanied by an upregulation of hypoxia-inducible factor subunit α (HIF1A) transcripts in the muscle cells. However, in the brain cells, HIF1A mRNA levels were suppressed during reoxygenation. Other functionally important transcripts and miRNAs involved in antioxidant response, apoptosis, inflammation, and substrate oxidation were also differentially expressed between the muscle and brain cells. Suppression of miRNA levels during acute intermittent hypoxia was stronger in the brain cells affecting ~ 55% of all studied miRNA transcripts than in the muscle cells (~ 25% of miRNA) signifying transcriptional derepression of the respective mRNA targets. Our study provides insights into the potential molecular and physiological mechanisms contributing to different hypoxia sensitivity of the studied tissues and can serve as a starting point to better understand the biological processes associated with hypoxia stress, e.g. during ischemia and reperfusion.


Syphilis and prostitution in the socio-medical geographies of Turkey's early republican provinces.
During and after the Ottoman Empire's collapse, Turkey's fledgling public health and social services ministry sought to deal with the increasing prevalence of syphilis-especially in its rural communities. This article examines the emergence of state-led information collection in Turkey during the 1920s and early 1930s and the anti-syphilis campaigns that resulted, and thus explores how the state created a new medical and moral order surrounding its citizens' sexualities that came to focus its gaze upon prostitution. Utilizing information from official primary sources, we analyze this transformation as part of a broader process of medicalization and state expansion that made syphilis a subject for state regulation. Within this context, moral pronouncements regarding the disease, traditional medicine, and prostitution and the potential benefits of regulated brothels were reframed, represented, and dispersed as directives for public health policy. Through this research, we assess how field-based surveys contributed ultimately to republican regimes of regulating sex work that still persist.


Alterations in endothelial cell proteinase and inhibitor polarized secretion following treatment with interleukin-1, phorbol ester, and human melanoma cell conditioned medium.
Polarized secretion of matrix metalloproteinases and plasminogen activators by monkey aortic endothelial cells was studied in vitro, using transwell inserts. The endothelial cells constitutively expressed matrix metalloproteinase-2, tissue inhibitors of metalloproteinases 1 and 2, urokinase, and tissue plasminogen activator, all with basal preference. Matrix metalloproteinase-9 activity was induced by phorbol 12-myristate 13-acetate (apical), interleukin-1 alpha (basal), and by conditioned medium from DX3 human melanoma cells (basal). The DX3 melanoma conditioned medium also stimulated basal secretion of matrix metalloproteinase-2, urokinase, tissue plasminogen activator, and tissue inhibitors of metalloproteinases. The rise in proteolytic activity in the basal direction was reflected by increased capacity to degrade subendothelial basement membrane type IV collagen, shown immunohistologically, using monkey kidney tissue sections and basement membrane deposited by endothelial cells into the transwell membrane. Thus, IL-1 alpha and DX3 melanoma conditioned medium can stimulate endothelial cells in vitro to concentrate secretion of proteinases spatially onto the underlying basement membrane. We suggest that the stimulation of endothelial cell proteinase activity by tumor cells may facilitate tumor cell extravasation.


Do integrated health care systems provide lower-cost, higher-quality care?
A study Looking at quality and cost issues for integrated vs. non integrated physician practices yields some interesting findings.


Atopy may be related to exercise-induced bronchospasm in asthma.
Recent studies suggest that atopy may be associated with exercise-induced bronchospasm (EIB) in asthma. However, it is not clear whether atopy is related to EIB, regardless of airway hyper-responsiveness (AHR) to methacholine, because asthmatic subjects often show AHR to exercise and methacholine simultaneously. To investigate whether atopy is related to EIB in asthmatic subjects, independently of AHR to methacholine. Fifty-eight male asthmatic subjects were studied. Initial spirometry was performed. Skin prick test was carried out, using 53 common allergens including mites dust antigen. Atopy score was defined as a sum of mean weal diameters to all allergens tested. Methacholine bronchial provocation testing was performed. Twenty-four hours later, free running test was performed. Positive EIB was defined as a 15% reduction or more in FEV1 from baseline after exercise. All subjects had AHR to methacholine. The degree of AHR to methacholine in asthmatics with EIB was similar to that in asthmatics without EIB. However, atopy score and skin reaction to Dermatophagoides pteronyssinus significantly increased in asthmatics with EIB compared with those without EIB (P < 0.05, respectively). Furthermore, the degree of EIB significantly correlated with atopy score in all subjects (r = 0.35, P < 0.01). This relationship was maintained even after the exclusion of EIB-negative asthmatic subjects. Atopy defined as skin test reactivity may contribute to the development of EIB in asthma, independently of AHR to methacholine.


Influence of municipal solid waste compost application on heavy metal content in soil.
Municipal solid waste composts (MSWC) are widely used over agricultural lands as organic soil amendment and fertilizer. However, MSWC use may result in various adverse impacts over agricultural lands. Especially, heavy metal contents of MSWC should always be taken into consideration while using in agricultural practices. The present study was conducted to find out heavy metal contents of municipal solid waste compost (MSWC) and to investigate their effects on soils. Experiments were carried out in three replications as field experiments for 2 years. Dry-based MSWC was applied to each plot at the ratios of 0, 40, 80, 120, 160, 200 t ha(-1). Results revealed that heavy metal content of MSWC was within the allowable legal limits. Compost treatments significantly increased Cu, Zn, Ni, Cr, Cd, and Pb content of soils (p < 0.01). Increasing soil heavy metal contents were observed with increasing applied compost doses. Although compost treatments significantly increased soil heavy metal contents, the final contents were still within the allowable legal limits. Results showed that MSWC doses over 10 t ha(-1) may create a heavy metal risk in long term for soils with pH ≥ 7. Therefore, in MSWC use over agricultural lands, heavy metal contents should always be taken into consideration and excessive uses should be avoided.


[Effect of the prolonged exposure to etylene glycol and fatty acid ester on the functional state of the body of experimental animals].
The effect of prolonged (for 1 1/2 month) action of ethylene glycol and fatty acids ester on the bodily function was studied in experiments on rats by means of a complex of techniques. The threshold dose determined in a subacute experiment amounted to 1.8625 g/kg. The cumulative effect of the preparation is evaluated as little and medium cumulative.


Regulation of the antiviral and anticellular activities of interferon by exogenous double-stranded RNA.
The effects of double-stranded RNA (dsRNA) on interferon (IFN)-induced antiviral and anticellular activities was investigated by introducing poly(I)-poly(C) into mouse L-cells. Coprecipitation of dsRNA with calcium phosphate enabled its efficient penetration into cells in culture. Rate of cellular protein synthesis was inhibited by dsRNA only in cultures pretreated with IFN. Moreover, the anticellular effect of IFN, as measured by the inhibition of cell DNA synthesis, was also enhanced by dsRNA. The kinetics of dsRNA-mediated inhibition of protein synthesis were relatively slow as compared with the inhibitory effect of 2'-5' oligoadenylic acid (2'5'A), which was also introduced into cells by the calcium phosphate coprecipitation technique. To analyze the effects of dsRNA on the antiviral state induced by IFN, vesicular stomatitis virus (VSV) and encephalomyocarditis virus (EMC), replications were followed by measuring viral-specific RNA synthesis in the cell. Introduction of dsRNA after the infection had no effect on VSV and EMC replication in control cells, and it enhanced, to a small extent, the antiviral state of cells pretreated with IFN. In contrast, introduction of 2'5'A into virus-infected cells inhibited VSV and EMC replications regardless of IFN pretreatment. This work demonstrated that the role of dsRNA in regulating the antiviral and anticellular activities of IFN could be studied by introducing exogenous dsRNA into cells in culture by the calcium phosphate coprecipitation technique.


Development of resistance in calves to nymphs of Rhipicephalus appendiculatus (Acarina:Ixodidae) during test feeds.
A test of host resistance to infestation by the tick Rhipicephalus appendiculatus was repeated 5 times, at weekly intervals, on 2 initially susceptible calves. One hundred nymphs (NN) were used each time. Pairs of tick-susceptible rabbits served as controls. The engorged NN were weighed, then allowed to moult individually, and the sex of each was noted after emergence of the adult. During the course of the experiment there was a significant reduction in mean engorged weight (especially in the pre-male NN) and also in the length of the feeding period, and a downward trend in the percentage of NN which engorged. There was a significant increase in the percentage of engorged NN that moulted. Pre-male NN were significantly heavier than pre-female NN after feeding on susceptible hosts, but though mean feeding times were the same, they took 1-2 days longer to moult than pre-females at 28 degrees C. It was concluded that the test itself could induce resistance if applied to fully susceptible calves.


Proprioceptive drift is affected by the intermanual distance rather than the distance from the body's midline in the rubber hand illusion.
In the rubber hand illusion (RHI), simultaneous brush stroking of a subject's hidden hand and a visible rubber hand induces a transient illusion of the latter to "feel like it's my hand" and a proprioceptive drift of the hidden own hand toward the rubber hand. Recent accounts of the RHI have suggested that the illusion would only occur if weighting of conflicting sensory information and their subsequent integration results in a statistically plausible compromise. In three different experiments, we investigated the role of distance between the two hands as well as their proximity to the body's midline in influencing the occurrence of the illusion. Overall, the results suggest that the illusion is abolished when placing the two hands apart, therefore increasing the mismatch between the visual and proprioceptive modality, whereas the proximity of the two hands to the body's midline plays only a minor role on the subjective report of the illusion. This might be driven by the response properties of visuotactile bimodal cells encoding the peripersonal space around the hand.


Molecular control of rodent spermatogenesis.
Spermatogenesis is a complex developmental process that ultimately generates mature spermatozoa. This process involves a phase of proliferative expansion, meiosis, and cytodifferentiation. Mouse models have been widely used to study spermatogenesis and have revealed many genes and molecular mechanisms that are crucial in this process. Although meiosis is generally considered as the most crucial phase of spermatogenesis, mouse models have shown that pre-meiotic and post-meiotic phases are equally important. Using knowledge generated from mouse models and in vitro studies, the current review provides an overview of the molecular control of rodent spermatogenesis. Finally, we briefly relate this knowledge to fertility problems in humans and discuss implications for future research. This article is part of a Special Issue entitled: Molecular Genetics of Human Reproductive Failure.


Controlling energy and charge transfer in linear chlorophyll dimers.
A series of linkers constructed from combinations of phenyl and ethynyl groups is shown to permit ultrafast energy transfer between two chlorophylls, while allowing control over radical cation migration between them.


A molecular basis for the two locus model of human complement component C4.
The major histocompatibility complex(MHC)-linked fourth component of complement (C4) shows a high degree of polymorphism in several animal species. In man C4 polymorphism was detected by distinct charge differences of the variants. O'Neill et al. showed that this C4 polymorphism was controlled by two closely linked genetic loci, F (C4A) and S (C4B) and these results were extended by Awdeh et al. with an improved typing method. Biochemical analysis of human C4 has revealed that it consists of three polypeptide chains, alpha, beta and gamma. In all reports so far on the molecular analysis of human C4, no molecular weight differences between the A and B locus-encoded molecules have been noticed. Here we demonstrate that the C4A and C4B locus-encoded alpha-chains have a molecular weight (MW) of 96,000 and 94,000, respectively, presenting for the first time a molecular basis for the difference between all C4A and C4B variants tested. Even rare variants that are difficult to allocate to the A or B locus on the basis of charge differences could be identified as C4A or C4B variants in this way, thereby providing new insights into the relationships between the C4A and C4B loci.


Quantifying mechanical heterogeneity in canine acute lung injury: impact of mean airway pressure.
The heterogeneous pattern of acute lung injury (ALI) predisposes patients to ventilator-associated lung injury. Currently, there is no simple technique that can reliably quantify lung heterogeneity during the dynamic conditions of mechanical ventilation. Such a technique may be of use in optimizing mechanical ventilatory parameters such as rate, tidal volume, or positive end-expiratory pressure. To determine the impact of heterogeneity on respiratory mechanics, the authors measured respiratory impedance (Zrs), expressed as respiratory resistance (Rrs) and elastance (Ers), in 11 anesthetized dogs from 0.078 to 8.9 Hz using broadband pressure and flow excitations under baseline conditions and after ALI produced by infusion of 0.08 ml/kg oleic acid into the right atrium. Data were obtained at mean airway pressures (Pao) of 5, 10, 15, and 20 cm H2O. The Zrs spectra were fit by various models of the respiratory system incorporating different distributions of parallel viscoelastic tissue properties. Under baseline conditions, both Rrs and Ers exhibited dependence on oscillation frequency, reflecting viscoelastic behavior. The Ers demonstrated significant dependence on Pao. After ALI, both the level and frequency dependence of Rrs and Ers increased, as well as the apparent heterogeneity of tissue properties. Both Rrs and Ers as well as heterogeneity decreased with increasing Pao, approaching baseline levels at the highest levels of Pao. These data demonstrate that Zrs can provide specific information regarding the mechanical heterogeneity of injured lungs at different levels of Pao. Moderate increases in Pao seem to be beneficial in ALI by reducing heterogeneity and recruiting lung units. These noninvasive measurements of lung heterogeneity may ultimately allow for the development of better ventilation protocols that optimize regional lung mechanics in patients with ALI.


Comparison of the sensitivity of Salmonella typhimurium strains YG1024 and YG1012 for detecting the mutagenicity of aromatic amines and nitroarenes.
Salmonella typhimurium YG1024 is a derivative of S. typhimurium TA98 with a high level of N-hydroxyarylamine O-acetyltransferase (OAT) activity. We have demonstrated that this strain is highly sensitive to the mutagenic actions of N-hydroxyarylamines derived from aromatic amines and nitroarenes. In this paper, we compared the sensitivities of YG1024 with those of S. typhimurium YG1012, which has about 4 times higher OAT activity than YG1024 but lacks plasmid pKM101. It turned out that YG1024 was more sensitive to the mutagenic actions of 1-aminonaphthalene, 1-nitropyrene, 1,8-dinitropyrene and 2-nitronaphthalene than YG1012 and showed comparable sensitivity to 2-hydroxy-acetylaminofluorene, 2-aminoanthracene and 2-amino-6-methyldipyrido[1,2-a:3',2'-d]imidazole (Glu-P-1) to YG1012. These results suggested that YG1024 is more suitable than YG1012 for the efficient detection of mutagenic aromatic amines and nitroarenes.


Response of diffuse sclerosing osteomyelitis of the mandible to alendronate: follow-up study by 99mTc scintigraphy.
We report a case of diffuse sclerosing osteomyelitis of the mandible responded to alendronate, after a poor response to intravenous antibiotics, antibiotic irrigation-perfusion, and decortication. The patient was given an intravenous infusion of 10mg of alendronate. Pain resolved within 24 h. There were no severe adverse events. Increased uptake of 99mTc in the mandible almost completely disappeared 3 months after treatment.


Independent statistical observables for ultrametric disordered populations.
It is not exceptional that a sample of N random data X(i), i=1,...,N contains ultrametric covariations, namely the matrix C with matrix elements <X(i)X(j)>-<X(i)><X(j)> is ultrametric. We define independent (decorrelated) "collective" observables by diagonalizing this matrix. Symmetry properties of such eigenvectors are discussed. Often also, however, while the existence of an ultrametric tree is known, the degrees of parentage of the data are unknown, because a random perturbation confuses the labeling of the leaves of the tree. We sort out those observables which are more robust with respect to such labeling mistakes.


Safety and efficiency of microwave ablation for recurrent and persistent secondary hyperparathyroidism after parathyroidectomy: A retrospective pilot study.
Recurrent and persistent secondary hyperparathyroidism (SHPT) nodules have an incidence of 10-70% after surgery. The treatment of recurrent and persistent SHPT nodules is a challenge, and surgical resection of difficult-to-reach or post-operative adhesions often fails. The aim of this research was to study the safety and effectiveness of microwave ablation (MWA) for recurrent and persistent SHPT. This was a retrospective study of 11 patients enrolled with a total of 16 nodules, and MWA was employed to manage SHPT. The laboratory test results, including the intact parathyroid hormone (iPTH), serum calcium, phosphorus and alkaline phosphatase (ALP) levels, improvement of SHPT-related symptoms after ablation, and complications during and after MWA were recorded and analysed. After ablation the value of iPTH was markedly decreased from 1570 ± 1765 pg/mL to 287 ± 239 pg/mL 1 day after MWA (p < 0.05). The levels of serum calcium and phosphorus decreased from 2.51 ± 0.23 mmol/L to 2.06 ± 0.27 mmol/L (p < 0.001) and 1.80 ± 0.43 mmol/L to 1.48 ± 0.32 mmol/L (p < 0.05), respectively, 1 day after MWA. There was no significant difference in the ALP value before and after MWA (p > 0.05). The clinical symptoms, including ostalgia, pruritus, disability, and restless legs, improved after MWA. Minor complications and side effects encountered during or after MWA include haematoma (1/11, 9%), transient hoarseness (2/11, 18.2%), hypocalcemia (6/11, 54.5%). No major complication occurred. MWA may be safe and effective to manage recurrent and persistent SHPT nodules; a definite conclusion needs to expand the sample size with a longer follow-up time.


Small molecule anticonvulsant agents with potent in vitro neuroprotection.
Severe seizure activity is associated with recurring cycles of excitotoxicity and oxidative stress that result in progressive neuronal damage and death. Intervention to halt these pathological processes is a compelling disease-modifying strategy for the treatment of seizure disorders. In the present study, a core small molecule with anticonvulsant activity has been structurally optimized for neuroprotection. Phenotypic screening of rat hippocampal cultures with nutrient medium depleted of antioxidants was utilized as a disease model. Increased cell death and decreased neuronal viability produced by acute treatment with glutamate or hydrogen peroxide were prevented by our novel molecules. The neuroprotection associated with this chemical series has marked structure activity relationships that focus on modification of the benzylic position of a 2-phenyl-2-hydroxyethyl sulfamide core structure. Complete separation between anticonvulsant activity and neuroprotective action was dependent on substitution at the benzylic carbon. Chiral selectivity was evident in that the S-enantiomer of the benzylic hydroxy group had neither neuroprotective nor anticonvulsant activity, while the R-enantiomer of the lead compound had full neuroprotective action at <40 nM and antiseizure activity in three animal models. These studies indicate that potent, multifunctional neuroprotective anticonvulsants are feasible within a single molecular entity.


Abdominal aortic pseudoaneurysm caused by prolonged methicillin-resistant Staphylococcus aureus sepsis.
The mechanism of pseudoaneurysm formation caused by prolonged sepsis is thought to be related to the vascular endothelium being directly invaded and broken by bacteria. Moreover, matrix metalloproteinases (MMPs) which are up-regulated by chronic inflammation have been reported to be implicated in the pathogenesis of aneurysm development through increased proteolysis of extracellular matrix proteins. An effective treatment for infected pseudoaneurysm remains unsettled. Surgery is generally performed, however, because the patients in most of these cases are in very poor physical condition, the operation is associated with high morbidity and mortality. A more successful alternative is endovascular treatment. Recent reports indicate low morbidity and mortality rates with this treatment. If the patient in this case had been in better condition, we could have selected endovascular stent-grafting for her treatment.


Pharmacokinetics and safety of a single dose of the novel LAMA/LABA fixed-dose combination of glycopyrronium/formoterol fumarate dihydrate metered dose inhaler, formulated using co-suspension delivery technology, in Japanese healthy subjects.
Chronic obstructive pulmonary disease (COPD) causes significant mortality in Japan. GFF MDI is a long-acting muscarinic antagonist/long-acting β2-agonist fixed-dose combination of glycopyrronium (GP) and formoterol fumarate dihydrate (FF), delivered by a metered dose inhaler (MDI) using co-suspension delivery technology, for the long-term maintenance treatment of COPD. This randomized, Phase I, single-dose, four-treatment, four-period, crossover study (NCT02196714) examined the pharmacokinetic (PK) and safety profile of two doses of GFF MDI (28.8 μg/10 μg and 14.4 μg/10 μg) and two doses of GP MDI (28.8 μg and 14.4 μg), both formulated using co-suspension delivery technology, in healthy Japanese subjects (18-45 years of age). PK parameters included area under the curve (AUC) from 0 to 12 h (AUC0-12), AUC from 0 to the time of the last measurable plasma concentration, maximum observed plasma concentration (Cmax), and time to Cmax. Safety was monitored throughout the study. Plasma GP profiles were comparable between GFF MDI and GP MDI formulations containing the same GP dose. Increases in GP AUC0-12 and Cmax were generally dose proportional from 14.4 to 28.8 μg after administration of either formulation. The addition of FF 10 μg to GP MDI 28.8 μg or 14.4 μg in a fixed-dose combination did not appreciably alter the PK of GP, nor did an increase in GP dose from 14.4 μg to 28.8 μg in a fixed-dose combination with FF 10 μg appreciably alter the PK of formoterol. Both formulations of GFF MDI and GP MDI were well tolerated in healthy Japanese subjects. Data from this study support further evaluation of GFF MDI in Japanese patients with COPD.


Dopaminergic stimulation of HGH in diabetes mellitus and in obesity.
The behavior of HGH in basal conditions and after L-Dopa infusion was studied in a group of patients with diabetic retinopathy, in 9 obese and 8 control subjects. In both diabetics and obese subjects, increases found in HGH plasma levels after administration of the drug were slighter than in the controls. On the basis of these results it can be concluded that in diabetics the availability of energetic substrates may modify the HGH response to the L-Dopa stimulus. In obesity, the possibility is considered of a reversible response to the L-Dopa stimulus. In obesity, the possibility is considered of a reversible defect in the sensitivity of the dopaminergic receptors, induced by metabolic and endocrine factors.


Evaluation of iron status in women identified by copper sulfate screening as ineligible to donate blood.
Assessment of venous hemoglobin (Hb), serum ferritin, and zinc protoporphyrin (ZPP) levels was carried out in women identified by CuSO4 screening as ineligible to donate blood. The correlation of log ferritin with ZPP was relatively poor (r = -0.580) but significant (p greater than 0.01). However, a ZPP level of 2.0 micrograms per g Hb or greater (upper limit of normal for first-time female donors) showed a predictive value of 0.85 for a ferritin level of 12 ng per ml or less in these donors. The correlation of hemoglobin concentration with ZPP level was significant (r = -0.667; p less than 0.001) in blood donors with ferritin levels at or below 12 ng per ml. ZPP level appears to be increased in iron-depleted (hypoferritinemic) blood donor in whom animals had developed or was developing (Hb less than 12.5 g/dl). Although direct measurements of venous hemoglobin and ferritin levels most accurately evaluate such blood donors, these tests are time-consuming and expensive and are currently not adaptable to bloodmobile operations. Copper sulfate screening has proved feasible in the bloodmobile setting, and the measurement of ZPP level has been used for mobile screening for lead poisoning. ZPP may be an inexpensive and useful screening test to determine a subset of donors who should receive supplemental iron or reduce their frequency of blood donation.


The sensory neurons of touch.
The somatosensory system decodes a wide range of tactile stimuli and thus endows us with a remarkable capacity for object recognition, texture discrimination, sensory-motor feedback and social exchange. The first step leading to perception of innocuous touch is activation of cutaneous sensory neurons called low-threshold mechanoreceptors (LTMRs). Here, we review the properties and functions of LTMRs, emphasizing the unique tuning properties of LTMR subtypes and the organizational logic of their peripheral and central axonal projections. We discuss the spinal cord neurophysiological representation of complex mechanical forces acting upon the skin and current views of how tactile information is processed and conveyed from the spinal cord to the brain. An integrative model in which ensembles of impulses arising from physiologically distinct LTMRs are integrated and processed in somatotopically aligned mechanosensory columns of the spinal cord dorsal horn underlies the nervous system's enormous capacity for perceiving the richness of the tactile world.


Reversal of dopamine neurons and locomotor ability degeneration in aged rats with smilagenin.
The purpose of this paper is to study the effect of smilagenin (SMI) (PYM50028), a sapogenin compound originally identified from Chinese medicinal herb, on dopamine neurons and locomotor ability in aged rats. Experiments were carried out on young and aged Sprague-Dawley rats, which were daily administered with either SMI (18mg/kg/day) or vehicle (0.5% sodium carboxymethycellulose [CMCNa]). Open-field and rotarod performance tests revealed that behavioral ability was impaired in aged rats and was improved by oral administration of smilagenin. Immunohistochemical data showed that tyrosine hydroxylase (TH)-positive neuron numbers in the substantia nigra pars compacta (unbiased stereological counting) were altered with aging and were increased by smilagenin treatment. Likewise, the dopamine receptor density and the striatal dopamine transporter (DAT) density ((125)I-2β-carbomethoxy-3β-(4-iodophenyl)-N-(3-fluoropropyl) nortropane [(125)I-FP-CIT] autoradiography) were significantly lowered in aged rats as compared to young rats, and treatment with smilagenin significantly elevated the dopamine receptor and DAT density in aged rats. Furthermore, smilagenin enhances glial cell-derived neurotrophic factor (GDNF) release both in the striatum and midbrain. These results indicate a possible role of smilagenin in the treatment of age-related extrapyramidal disorders.


Confirmed E-cigarette or vaping product use associated lung injury (EVALI) with lung biopsy; A case report and literature review.
E-cigarette or vaping has become an increasingly popular alternative to smoking tobacco. In September 2019 multiple cases of confirmed E-cigarette or vaping product use associated lung injury were published. However, there is limited knowledge regarding the pathologic mechanism of this condition. We performed a systematic literature review in PubMed and EMBASE aiming to obtain additional clinical data on confirmed E-cigarette or vaping product use associated lung injury cases with lung biopsy results. With this information we hope to determine whether this condition is related to a histopathological pattern of acute lung injury instead of lipid deposits. Seven articles were reviewed and a total of 27 cases were included. Imaging findings predominantly showed presence of diffuse bilateral ground glass opacities. A majority of patients had complete resolution of the disease. The most common histopathological pattern was organizing pneumonia present in almost half of the patients. Other frequently occurring patterns included diffuse alveolar damage and acute fibrinous pneumonitis; lipoid pneumonia was found in one case. The underlying pathophysiological mechanism in E-cigarette or vaping product use associated lung injury is most likely acute lung injury related to direct inhalant-mediated parenchymal inflammation.


A chromosome-level genome assembly for the astaxanthin-producing microalga Haematococcus pluvialis.
The green microalga Haematococcus pluvialis can synthesize high amounts of astaxanthin, which is a valuable antioxidant that has been utilized in human health, cosmetics, and aquaculture. To illustrate detailed molecular clues to astaxanthin yield, we performed PacBio HIFI along with Hi-C sequencing to construct an improved chromosome-level haplotypic genome assembly with 32 chromosomes and a genome size of 316.0 Mb. Its scaffold N50 (942.6 kb) and contig N50 (304.8 kb) have been upgraded remarkably from our previous genome draft, and a total of 32,416 protein-coding genes were predicted. We also established a high-evidence phylogenetic tree from seven representative algae species, with the main aim to calculate their divergence times and identify expanded/contracted gene families. We also characterized genome-wide localizations on chromosomes of some important genes such as five BKTs (encoding beta-carotene ketolases) that are putatively involved in astaxanthin production. In summary, we reported the first chromosome-scale map of H. pluvialis, which provides a valuable genetic resource for in-depth biomedical investigations on this momentous green alga and commercial astaxanthin bioproduction.


A Begomovirus DNAbeta-encoded protein binds DNA, functions as a suppressor of RNA silencing, and targets the cell nucleus.
Our previous results demonstrated that the DNAbeta satellite (Y10beta) associated with Tomato yellow leaf curl China virus Y10 isolate (TYLCCNV-Y10) is essential for induction of leaf curl symptoms in plants and that transgenic expression of its betaC1 gene in Nicotiana plants induces virus-like symptoms. In the present study, in vitro DNA binding activity of the betaC1 proteins of Y10beta and DNAbeta (Y35beta) found in the Tobacco curly shoot virus Y35 isolate (TbCSV-Y35) were studied following their expression as six-His fusion proteins in Escherichia coli. Electrophoretic mobility shift assays and UV cross-linking experiments revealed that betaC1 proteins could bind both single-stranded and double-stranded DNA without size or sequence specificity. Suppression of green fluorescent protein (GFP) transgene silencing was observed with the new leaves of GFP-expressing Nicotiana benthamiana plants coinoculated by TYLCCNV-Y10 plus Y10beta or by TbCSV-Y35 plus Y35beta. In a patch agroinfiltration assay, the transiently expressed betaC1 gene of Y10beta or Y35beta was able to suppress host RNA silencing activities and permitted the accumulation of high levels of GFP mRNA in the infiltrated leaf patches of GFP transgenic N. benthamiana plants. The betaC1 protein of Y10beta accumulated primarily in the nuclei of plant and insect cells when fused with beta-glucuronidase or GFP and immunogold labeling showed that the betaC1 protein is present in the nuclei of infected N. benthamiana plants. A mutant version of Y10beta carrying the mutations within the putative nuclear localization sequence of the Y10 betaC1 protein failed to induce disease symptoms, suppress RNA silencing, or accumulate in the nucleus, suggesting that nuclear localization of the betaC1 protein is a key requirement for symptom induction and silencing suppression.


Quantitative microbial risk assessment associated with ready-to-eat salads following the application of farmyard manure and slurry or anaerobic digestate to arable lands.
Farmyard manure and slurry (FYM&S) and anaerobic digestate are potentially valuable soil conditioners providing important nutrients for plant development and growth. However, these organic fertilisers may pose a microbial health risk to humans. A quantitative microbial risk assessment (QMRA) model was developed to investigate the potential human exposure to pathogens following the application of FYM&S and digestate to agricultural land. The farm-to-fork probabilistic model investigated the fate of microbial indicators (total coliforms and enterococci) and foodborne pathogens in the soil with potential contamination of ready-to-eat salads (RTEs) at the point of human consumption. The processes examined included pathogen inactivation during mesophilic anaerobic digestion (M-AD), post-AD pasteurisation, storage, dilution while spreading, decay in soil, post-harvest washing processes, and finally, the potential growth of the pathogen during refrigeration/storage at the retail level in the Irish context. The QMRA highlighted a very low annual probability of risk (Pannual) due to Clostridium perfringens, norovirus, and Salmonella Newport across all scenarios. Mycobacterium avium may result in a very high mean Pannual for the application of raw FYM&S, while Cryptosporidium parvum and pathogenic E. coli showed high Pannual, and Listeria monocytogenes displayed moderate Pannual for raw FYM&S application. The use of AD reduces this risk; however, pasteurisation reduces the Pannual to an even greater extent posing a very low risk. An overall sensitivity analysis revealed that mesophilic-AD's inactivation effect is the most sensitive parameter of the QMRA, followed by storage and the decay on the field (all negatively correlated to risk estimate). The information generated from this model can help to inform guidelines for policymakers on the maximum permissible indicator or pathogen contamination levels in the digestate. The QMRA can also provide the AD industry with a safety assessment of pathogenic organisms resulting from the digestion of FYM&S.


Outcomes for general medical inpatients with diabetes mellitus and new hyperglycaemia.
To investigate the relationship between admission glycaemic status and inpatient mortality in patients with and without pre-existing diabetes. Prospective observational cohort study. A general medical ward in an Australian tertiary referral hospital. 903 patients admitted to the general medical ward between February 2003 and July 2004. Inpatient death. The overall inpatient mortality was 5.4% (n = 49). In the total cohort, age > 75 years and admission fasting plasma glucose (FPG) levels > or = 5.6 mmol/L were independent predictors of mortality. For patients without a known history of diabetes, each 1 mmol/L rise in admission FPG was associated with a 33% increase in mortality. In these patients, elevated (> 6.0%) and normal glycated haemoglobin (HbA(1c)) levels were associated with mortalities of 11.3% and 4.4%, respectively (odds ratio, 2.47; 95% CI, 1.16-5.26). In contrast, in patients with known diabetes, there was no association between admission FPG levels, HbA(1c) and mortality. Length of stay was not independently associated with FPG, HbA(1c), or diabetes status. In patients without known diabetes, the risk of death was increased for admission FPG levels > or = 5.6 mmol/L. However, pre-existing abnormal glucose metabolism, reflected by elevated HbA(1c) levels, appeared a more important predictor of inpatient mortality than glucose levels in patients without known diabetes.


Exosomal-microRNA transcriptome profiling of Parental and CSC-like MDA-MB-231 cells in response to cisplatin treatment.
Triple negative breast cancer (TNBC) is the most aggressive breast cancer subtype with higher risk of metastasis and cancer reoccurrence. Cisplatin is one of the potential anticancer drugs for treating TNBC, where its effectiveness remains challenged by frequent occurrence of cisplatin resistance. Since acquirement of drug resistance often being associated with presence of cancer stem cells (CSCs), investigation has been conducted, suggesting CSC-like subpopulation to be more resistant to cisplatin than their parental counterpart. On the other hand, plethora evidences showed the transmission of exosomal-miRNAs are capable of promoting drug resistance in breast cancers. In this study, we aim to elucidate the differential expression of exosomal-microRNAs profile and reveal the potential target genes in correlation to cisplatin resistance associated with CSC-like subpopulation by using TNBC cell line (MDA-MB-231). Utilizing next generation sequencing and Nanostring techniques, cisplatin-induced dysregulation of exosomal-miRNAs were evaluated in maximal for CSC-like subpopulation as compared to parental cells. Intriguingly, more oncogenic exosomal-miRNAs profile was detected from treated CSC-like subpopulation, which may correlate to enhancement of drug resistance and maintenance of CSCs. In treated CSC-like subpopulation, unique clusters of exosomal-miRNAs namely miR-221-3p, miR-196a-5p, miR-17-5p and miR-126-3p were predicted to target on six genes (ATXN1, LATS1, GSK3β, ITGA6, JAG1 and MYC), aligned with previous finding which demonstrated dysregulation of these genes in treated CSC-like subpopulation. Our results highlight the potential correlation of exosomal-miRNAs and their target genes as well as novel perspectives of the corresponding pathways that may be essential to contribute to the attenuated cytotoxicity of cisplatin in CSC-like subpopulation.


3D Dental Subsurface Imaging Using Enhanced Truncated Correlation-Photothermal Coherence Tomography.
Development of accurate and sensitive dental imaging technologies is a top priority in the pursuit of high-quality dental care. However, while early dental caries detection and routine monitoring of treatment progress are crucial for effective long-term results, current radiographic technologies fall short of this objective due to low sensitivity for small lesions and use of ionizing radiation which is unsuitable for frequent monitoring. Here we demonstrate the first application of enhanced Truncated Correlation-Photothermal Coherence Tomography (eTC-PCT) to dental imaging. eTC-PCT is non-invasive and non-ionizing, operates well below the maximum permissible exposure (MPE) limit, and features 3D subsurface imaging capability with operator controlled axial resolution. We explore the potential of this method for dental applications and demonstrate its capability for depth-resolved tomographic 3D reconstructions of the details and subsurface extent of a variety of dental defects. To this end, in this proof-of-concept study, dental eTC-PCT imaging results, and its sensitivity to dental caries, are discussed in comparison with visual examination, x-rays and micro-CT imaging.


Cytochrome p450 2C inhibition reduces post-ischemic vascular dysfunction.
Cytochrome p450 (CYP) inhibitors provide protection against myocardial infarction following both global and focal cardiac ischemia and reperfusion (I/R). We hypothesized that sulfaphenazole, an inhibitor of CYP2C6 and 9, also attenuates post-ischemic endothelial dysfunction by reducing CYP-mediated superoxide generation (which scavenges nitric oxide (NO)), thereby restoring NO bioavailability and vascular tone. Rat hearts were perfused in the Langendorff mode for 20 min in the presence, or absence, of sulfaphenazole and then subjected to 30 min global no-flow ischemia followed by 15 min reperfusion. Septal coronary resistance arteries were isolated and mounted on glass cannulae for measurements of luminal diameter. Preconstricted arteries were exposed to acetylcholine to elicit endothelium-dependent, NO-mediated vasodilation. Acetylcholine caused near maximal dilation in control tissues not subjected to I/R. Following I/R, endothelium-dependent vasodilation was reduced. Pretreatment with sulfaphenazole restored endothelial sensitivity to acetylcholine. Vasoresponsiveness to endothelium-independent vasodilators, sodium nitroprusside and isoproterenol, were also reduced following I/R. However, sensitivity to endothelium-independent vasodilators was not restored by pretreatment with sulfaphenazole. I/R-induced superoxide production was assessed by dihydroethidium staining of flash frozen hearts. Sulfaphenazole treatment significantly reduced superoxide production in arterial walls following I/R injury. We conclude that sulfaphenazole restores post-ischemic endothelium-dependent, NO-mediated vasodilation by reducing superoxide production, suggesting that CYP2C9 plays a key role in post-ischemic vascular dysfunction.


Residency-coordinator perceptions of psychiatry residency candidates: a pilot study.
The authors sought to assess whether Residency Training Coordinators (RTCs) can predict psychiatry residency applicants' success in obtaining a residency position. RTCs completed a rating form to measure their perceptions of the residency candidates' attentiveness, communication, attitude, and professionalism and answered the question "Would you feel comfortable seeing this person as your doctor?" These data were paired with ranking data from the National Residency Matching Program. Residency candidates rated highly by the coordinators were more likely to be ranked by Southern Illinois University (SIU) match at SIU. A "No" answer to the question "Would you feel comfortable seeing this person as your doctor?" was especially predictive of failure to match. RTCs can provide an important perspective on residency candidates' attentiveness, communication, attitude, and professionalism. Asking RTCs the question "Would you feel comfortable seeing this person as your doctor?" provides a simple and powerful screening tool.


Reduction in Clinical Variance Using Targeted Design Changes in Computerized Provider Order Entry (CPOE) Order Sets: Impact on Hospitalized Children with Acute Asthma Exacerbation.
Unwarranted variance in healthcare has been associated with prolonged length of stay, diminished health and increased cost. Practice variance in the management of asthma can be significant and few investigators have evaluated strategies to reduce this variance. We hypothesized that selective redesign of order sets using different ways to frame the order and physician decision-making in a computerized provider order entry system could increase adherence to evidence-based care and reduce population-specific variance. The study focused on the use of an evidence-based asthma exacerbation order set in the electronic health record (EHR) before and after order set redesign. In the Baseline period, the EHR was queried for frequency of use of an asthma exacerbation order set and its individual orders. Important individual orders with suboptimal use were targeted for redesign. Data from a Post-Intervention period were then analyzed. In the Baseline period there were 245 patient visits in which the acute asthma exacerbation order set was selected. The utilization frequency of most orders in the order set during this period exceeded 90%. Three care items were targeted for intervention due to suboptimal utilization: admission weight, activity center use and peak flow measurements. In the Post-Intervention period there were 213 patient visits. Order set redesign using different default order content resulted in significant improvement in the utilization of orders for all 3 items: admission weight (79.2% to 94.8% utilization, p<0.001), activity center (84.1% to 95.3% utilization, p<0.001) and peak flow (18.8% to 55.9% utilization, p<0.001). Utilization of peak flow orders for children ≥8 years of age increased from 42.7% to 94.1% (p<0.001). Details of order set design greatly influence clinician prescribing behavior. Queries of the EHR reveal variance associated with ordering frequencies. Targeting and changing order set design elements in a CPOE system results in improved selection of evidence-based care.


Potential anticancer agents IX. Isolation of a new simaroubolide, 6alpha-tigloyloxychaparrinone, from Ailanthus integrifolia ssp. calycina (Simaroubaceae).
Chaparrinone (1) and 6alpha-tigloyoxychaparrinone (2) were shown to be responsible for the antitumor and cytotoxic activities of the root bark of Ailanthus integrifolia ssp. calycina. The structure of the latter compound was established by analysis of spectral data. Compound 2 exhibited a more pronounced biological activity than chaparrinone (1) and demonstrates, for the first time, anticancer activity of a simaroubolide ester substituted at only the 6-position.


Hepatoatrial anastomosis and tricuspid valve replacement in Budd-Chiari syndrome 8.
The rare complication of tricuspid valve stenosis due to displacement of a peritoneovenous shunt catheter developed in a patient with Budd-Chiari syndrome. Surgical management included shunt removal and tricuspid valve replacement combined with hepatoatrial anastomosis. The tricuspid valve was replaced with a bioprosthesis, and liver resection opened the stenotic confluence of the hepatic veins. The right atrium was sutured to the liver capsule, thereby restoring physiologic hepatic venous blood flow.


The efficacy of dispersal in relation to safe site area and seed production.
A model is developed to define the efficacy of dispersal in relation to safe site area and seed production. Efficacy is measured as the expected number of progeny from one parent. It is shown that maximization of efficacy does not depend on the density of safe sites. When safe sites are confined to a restricted area around the parent, and safe sites are small or few propagules are produced, dispersal curves with short tails are most efficacious; when safe sites are larger or when more propagules are produced, distributions with longer tails become more advantageous.


The coccidia of sheep and goats in Senegal.
A survey of the coccidia in domestic sheep and goats was undertaken to ascertain the type of Eimeria species and the number of different coccidial species in individual faecal samples. Simultaneously the prevalence and the oocyst output was investigated in 2234 sheep and 577 goats during a 12-month period. Eight Eimeria species were encountered in sheep: E. ahsata, E. crandallis, E. faurei, E. intricata, E. ovina, E. ovinoidalis, E. pallidas and E. parva. In goats the following species were found: E. ahsata, E. arloingi, E. christenseni, E. crandallis, E. faurei, E. intricata, E. ninakohlyakimovae and E. parva. The prevalence in sheep ws 94% and in goats 85%, multiple parasitism was the rule. No seasonal fluctuation was observed in the prevalence or oocyst output. The sheep and goats' oocyst output was moderate, the mean for both being in the range of 1000-5000 oocysts/g of faeces.


Salvage of both lower extremities using simultaneous bilateral gastrocnemius and soleus muscle flaps: a case report.
We present a case of bilateral lower limb salvage in a young man with extensively exposed tibial bone following thermal injury. The medial head of the respective gastrocnemius muscle was used to cover the proximal third of debrided tibia, and the soleus muscle was used for the middle third coverage of each leg. This provided adequate, stable soft-tissue coverage for full recovery of both extremities and sufficient residual function for unassisted ambulation six months postoperatively.


Nickel-catalyzed skeletal transformation of tropone derivatives via C-C bond activation: catalyst-controlled access to diverse ring systems.
We report herein on nickel-catalyzed carbon-carbon bond cleavage reactions of 2,4,6-cycloheptatrien-1-one (tropone) derivatives. When a Ni/N-heterocyclic carbene catalyst is used, decarbonylation proceeds with the formation of a benzene ring, while the use of bidentate ligands in conjunction with an alcohol additive results in a two-carbon ring contraction with the generation of cyclopentadiene derivatives. The latter reaction involves a nickel-ketene complex as an intermediate, which was characterized by X-ray crystallography. The choice of an appropriate ligand allows for selective synthesis of four different products via the cleavage of a seven-membered carbocyclic skeleton. Reaction mechanisms and ligand-controlled selectivity for both types of ring contraction reactions were also investigated computationally.


Synthesis and structure of a hexacoordinate carbon compound.
In an exploration of six coordination and hypervalence in carbon compounds, steric constraints have been employed to bring four ether O atoms in close proximity to an allenic carbon atom. The dimethylated dication 2 is confirmed to have hexacoordinate carbon by experimental charge density analysis and DFT calculations and is arguably hypervalent.


Diagnostic performance of HMGA2 gene expression for differentiation of malignant thyroid nodules: A systematic review and meta-analysis.
The purpose of the current study was to investigate the diagnostic performance of high mobility group A2 (HMGA2) gene expression for differentiation of malignant thyroid nodules through a systematic review and meta-analysis. The MEDLINE/PubMed and EMBASE database, from the earliest available date of indexing through 10 April 2018, were searched for studies evaluating the diagnostic performance of HMGA2 expression for differentiation of thyroid nodules. We determined the sensitivities and specificities across studies, calculated positive and negative likelihood ratios (LR+ and LR-), and constructed summary receiver operating characteristic (ROC) curves. Across 7 studies, the pooled sensitivity for HMGA2 expression was 0.78 (95% CI; 0.67-0.86) with heterogeneity (I2 = 86.6) and a pooled specificity of 0.94 (95% CI: 0.85-0.98) with heterogeneity (I2 = 94.7). Likelihood ratio (LR) syntheses gave an overall positive likelihood ratio (LR+) of 12.6 (95% CI: 5.1-31.3) and negative likelihood ratio (LR-) of 0.24 (95% CI: 0.15-0.36). The pooled diagnostic odds ratio (DOR) was 53 (95% CI: 18-159). Hierarchical summary ROC curve indicates that the areas under the curve were 0.92 (95% CI: 0.89-0.94). In meta-regression analysis, no definite variable was the source of the study heterogeneity. The current meta-analysis showed the moderate sensitivity and high specificity of HMGA2 expression for differentiation of malignant thyroid nodules. The likelihood ratio scatter-gram suggested that HMGA2 expression analysis could be useful for confirmation of the presence of malignant thyroid nodules. Considering the heterogeneity of included studies, further large prospective studies are necessary to confirm these results.


Contrasting whole-genome and reduced representation sequencing for population demographic and adaptive inference: an alpine mammal case study.
Genomes capture the adaptive and demographic history of a species, but the choice of sequencing strategy and sample size can impact such inferences. We compared whole genome and reduced representation sequencing approaches to study the population demographic and adaptive signals of the North American mountain goat (Oreamnos americanus). We applied the restriction site-associated DNA sequencing (RADseq) approach to 254 individuals and whole genome resequencing (WGS) approach to 35 individuals across the species range at mid-level coverage (9X) and to 5 individuals at high coverage (30X). We used ANGSD to estimate the genotype likelihoods and estimated the effective population size (Ne), population structure, and explicitly modelled the demographic history with δaδi and MSMC2. The data sets were overall concordant in supporting a glacial induced vicariance and extremely low Ne in mountain goats. We evaluated a set of climatic variables and geographic location as predictors of genetic diversity using redundancy analysis. A moderate proportion of total variance (36% for WGS and 21% for RADseq data sets) was explained by geography and climate variables; both data sets support a large impact of drift and some degree of local adaptation. The empirical similarities of WGS and RADseq presented herein reassuringly suggest that both approaches will recover large demographic and adaptive signals in a population; however, WGS offers several advantages over RADseq, such as inferring adaptive processes and calculating runs-of-homozygosity estimates. Considering the predicted climate-induced changes in alpine environments and the genetically depauperate mountain goat, the long-term adaptive capabilities of this enigmatic species are questionable.


Rapid and nondestructive determination of sesamin and sesamolin in Chinese sesames by near-infrared spectroscopy coupling with chemometric method.
Sesame was one of the most important crops in Africa and east Asia. The sesamin and sesamolin in sesames have shown various pharmacological, biological and physiologic activities. In this study, a rapid and nondestructive method for determination of sesamin and sesamolin in Chinese sesames by near-infrared spectroscopy coupled with chemometric method was proposed. The near infrared spectra of sesame samples from three different Chinese areas were collected and the partial least squares (PLS) was used to construct the quantitative models. The spectral preprocessing and variable selection methods were adopted to improve the predictability and stability of the model. Reasonable quantitative results can be obtained when the samples used for model construction and prediction were harvested in same years. For sesamin and sesamolin, the correlation coefficient (R) and root mean square error prediction (RMSEP) were 0.9754, 0.9636 and 151.2951, 39.7720, respectively. The optimized models seem less effective when they were used to predict the samples harvested in other years or countries. However, acceptable results can still be obtained.


Comprehensive de novo mutation discovery with HiFi long-read sequencing.
Long-read sequencing (LRS) techniques have been very successful in identifying structural variants (SVs). However, the high error rate of LRS made the detection of small variants (substitutions and short indels < 20 bp) more challenging. The introduction of PacBio HiFi sequencing makes LRS also suited for detecting small variation. Here we evaluate the ability of HiFi reads to detect de novo mutations (DNMs) of all types, which are technically challenging variant types and a major cause of sporadic, severe, early-onset disease. We sequenced the genomes of eight parent-child trios using high coverage PacBio HiFi LRS (~ 30-fold coverage) and Illumina short-read sequencing (SRS) (~ 50-fold coverage). De novo substitutions, small indels, short tandem repeats (STRs) and SVs were called in both datasets and compared to each other to assess the accuracy of HiFi LRS. In addition, we determined the parent-of-origin of the small DNMs using phasing. We identified a total of 672 and 859 de novo substitutions/indels, 28 and 126 de novo STRs, and 24 and 1 de novo SVs in LRS and SRS respectively. For the small variants, there was a 92 and 85% concordance between the platforms. For the STRs and SVs, the concordance was 3.6 and 0.8%, and 4 and 100% respectively. We successfully validated 27/54 LRS-unique small variants, of which 11 (41%) were confirmed as true de novo events. For the SRS-unique small variants, we validated 42/133 DNMs and 8 (19%) were confirmed as true de novo event. Validation of 18 LRS-unique de novo STR calls confirmed none of the repeat expansions as true DNM. Confirmation of the 23 LRS-unique SVs was possible for 19 candidate SVs of which 10 (52.6%) were true de novo events. Furthermore, we were able to assign 96% of DNMs to their parental allele with LRS data, as opposed to just 20% with SRS data. HiFi LRS can now produce the most comprehensive variant dataset obtainable by a single technology in a single laboratory, allowing accurate calling of substitutions, indels, STRs and SVs. The accuracy even allows sensitive calling of DNMs on all variant levels, and also allows for phasing, which helps to distinguish true positive from false positive DNMs.


Genomic characterization and expression analysis of four apolipoprotein A-IV paralogs in Senegalese sole (Solea senegalensis Kaup).
The apolipoprotein A-IV (ApoA-IV) plays a key role in lipid transport and feed intake regulation. In this work, four cDNA sequences encoding ApoA-IV paralogs were identified. Sequence analysis revealed conserved structural features including the common 33-codon block and nine repeated motifs. Gene structure analysis identified four exons and three introns except for apoA-IVAa1 (with only 3 exons). Synteny analysis showed that the four paralogs were structured into two clusters (cluster A containing apoA-IVAa1 and apoA-IVAa2 and cluster B with apoA-IVBa3 and apoA-IVBa4) linked to an apolipoprotein E. Phylogenetic analysis clearly separated the paralogs according to their cluster organization as well as revealed four subclades highly conserved in Acanthopterygii. Whole-mount analyses (WISH) in early larvae (0 and 1day post-hatch (dph)) showed that the four paralogs were mainly expressed in yolk syncytial layer surrounding the oil globules. Later, at 3 and 5dph, the four paralogs were mainly expressed in liver and intestine although with differences in their relative abundance and temporal expression patterns. Diet supply triggered the intensity of WISH signals in the intestine of the four paralogs. Quantification of mRNA abundance by qPCR using whole larvae only detected the induction by diet at 5dph. Moreover, transcript levels increased progressively with age except for apoA-IVAa2, which appeared as a low-expressed isoform. Expression analysis in juvenile tissues confirmed that the four paralogs were mainly expressed in liver and intestine and secondary in other tissues. The role of these ApoA-IV genes in lipid transport and the possible role of apoA-IVAa2 as a regulatory form are discussed.


Characterization of polyamide-6/ propolis blended electrospun fibers.
Polyamide-6 (PA-6) nanofibers and PA-6/propolis ethanolic extract (EEP) blended fibers were prepared having electrospun their solutions in formic acid as solvent. The effect of concentrations of PA-6 and also EEP in polymer solutions on the morphology and physicochemical characteristics of their electrospun fibers was investigated. The analysis of FESEM images showed the mean diameter of fibers increased from 487- 682 nm with increasing PA-6 concentration in the range of 25-40 % w/v. While, increasing EEP concentration (20-50% (w/w)) in PA-6/EEP system caused the increasing fiber mean diameters from 943- 1773 nm. Partially high aspect ratio nanofibers were observed only in the PA-6 systems. Antioxidant activity of the fibers enhanced with increasing EEP concentration in the fiber mats. FTIR spectrums and thermal properties of electrospun fibers exhibited the simple mixtures of PA-6 and EEP in blend fibers which did not contain very complex interactions.


Current good manufacturing practice in plant automation of biological production processes.
The production of biologicals is subject to strict governmental regulations. These are drawn up in current good manufacturing practices (cGMP), a.o. by the U.S. Food and Drug Administration. To implement cGMP in a production facility, plant automation becomes an essential tool. For this purpose Manufacturing Execution Systems (MES) have been developed that control all operations inside a production facility. The introduction of these recipe-driven control systems that follow ISA S88 standards for batch processes has made it possible to implement cGMP regulations in the control strategy of biological production processes. Next to this, an MES offers additional features such as stock management, planning and routing tools, process-dependent control, implementation of software sensors and predictive models, application of historical data and on-line statistical techniques for trend analysis and detection of instrumentation failures. This paper focuses on the development of new production strategies in which cGMP guidelines are an essential part.


Determination of moniliformin using SAX column clean-up and HPLC/DAD-detection.
This work describes a method for the determination of theFusarium mycotoxin moniliformin (MON) in cereals. In addition to the optimization of the clean-up and the HPLC determination the most efficient extraction mode was investigated on natural contaminated samples. The method was validated for maize and wheat using a calibration range from 57 to 2300 µg/kg. Due to the ionic nature of the toxin the clean-up of the extracts was carried out with strong-anion-exchange columns. Moniliformin was separated by reversed phase ion-pair-chromatography (RP-Ion pair-HPLC) and detected by DAD. The validated method yielded recoveries of 76%±9% (maize) and 87%±5% (wheat) and detection limits of 39 µg/kg and 30 µg/kg, respectively. The suitability of the developed method was demonstrated on natural contaminated samples.


Effect of dietary oils enriched with n-3 fatty acids on survival of mice.
Female mice were fed a conventional diet, shifted at 119 days of age to a diet supplemented with 10 wt % lard (Lar), high-linoleic (n-6) safflower oil (Saf), rapeseed oil (low-erucic, Rap), high-alpha-linolenic (n-3) perilla oil (Per) or a mixture (1:9) of ethyl docosahexaenoate (n-3) and soybean oil (DHA/Soy). Weight gain was less in the Per group than in the other groups at 497 days of age. In the Rap group, proteinuria was more severe than in the Saf, Per and DHA/Soy group, and hepatic triacylglycerol accumulation was greater than in the other groups. The mean survival time of the DHA/Soy group (753 days) was significantly longer than in the Lar group (672 days) and Saf group (689 days); the differences among other groups (e.g., 701 days in the Per group and 712 days in the Rap group) were not statistically significant. Although DHA is more susceptible to auto-oxidation than other major fatty acids in the air, an oil containing DHA was found to increase the survival of mice. Rapeseed oil that decreases the survival time of SHRSP rats was found to be safe in the mouse strain used in this study when survival was an end point.


Magnetic resonance imaging in the diagnosis of a psoas abscess.
Identification of a psoas abscess is often a diagnostic challenge. In this case report the diagnosis and subsequent treatment of a psoas abscess previously suggested by computed tomography was greatly enhanced by magnetic resonance imaging.


Randomized, triple-blind, placebo-controlled study to evaluate the safety of 6'-Sialyllactose in healthy adults.
Sialyllactoses (SL) are an abundant component of human milk. There have been many studies on the biological effects of SL in humans. SL can be produced using an economical method of enzyme synthesis. Although the European Food Safety Authority has published the human safety and appropriate intake dose of 6'-SL sodium salt as a novel food, it has suggested that the appropriate dose for particular medical purposes be judged on a case-by-case basis. Also, as revealed in the same report, there are no data on toxicity when 6'-SL is used in human intervention. However, clinical studies have only confirmed the safety of 3'-SL for therapeutic intervention in humans, and the safety for therapeutic use of 6'-SL, which is more abundant than 3'-SL in human milk, has not been confirmed. In this study, to determine the safety of 6'-SL use in humans, participants were randomly assigned to the placebo (maltodextrin) and 6'-SL groups, and then 3 g of powder was orally administered twice a day for 12 weeks. There were no serious adverse reactions, such as life-threatening complications requiring hospitalization, causing disability, or causing deformity during the use of 6'-SL. There were no clinically significant differences among the baseline, sixth, and twelfth week clinical chemistry tests, such as aspartate aminotransferase, alanine aminotransferase, and creatinine. Most of the adverse reactions were gastrointestinal problems such as diarrhea, abdominal discomfort, and bloating, with no significant difference in the proportions between the placebo and 6'-SL groups. These results support the safety of the 6'-SL for human use.


Obsessive compulsive disorders in eating disorders.
The aim of this study is to explore current and lifetime prevalence of obsessive compulsive disorders (OCD) in eating disorder (ED) subgroups and subtypes defined by the DSM-IV and to study the chronology of appearance of these disorders taking into account the role played by denutrition. Current and lifetime prevalence were investigated using the Mini International Neuropsychiatric Interview (MINI) and the Yale-Brown Obsessive Compulsive Scale in a sample of 89 DSM-IV ED patients (58 AN and 31 BN) and 89 matched controls. Current and lifetime prevalence of OCD in ED was significantly higher than in general population (15.7% and 19% vs. 0% and 1.1%, P<.05). Anorexic patients presented a slightly higher current and lifetime comorbidity than bulimic patients (19% and 22.4% vs. 9.7% and 12.9%, n.s.). Purging anorexia was the diagnostic subtype, which presented the higher prevalences (29% and 43%), followed by restrictive anorexia (16%) and purging bulimia (13%). In the great majority of cases (65%), OCD diagnosis preceded ED diagnosis. Finally, OCD current prevalence and Y-BOCS scores of underweight patients were not significantly higher than normal-weight patients, suggesting that there were only limited links between denutrition and obsessionality.


"Ranaviruses: an emerging threat to ectothermic vertebrates" report of the First International Symposium on Ranaviruses, Minneapolis MN July 8, 2011.
This is a report of the First International Symposium on Ranaviruses held on July 8, 2011 in conjunction with the annual Joint Meeting of Ichthyologists and Herpetologists (JMIH) in Minneapolis, Minnesota, USA. The emerging threat of ranavirus infectious diseases to the global biodiversity of ectothermic vertebrates was addressed by 23 scientists from nine countries with expertise in ecology, pathology, virology, veterinary medicine and immunology.


Specifying the domain-general resources that contribute to conceptual construction: Evidence from the child's acquisition of vitalist biology.
There are two dissociable processes that underlie knowledge acquisition: knowledge enrichment, which involves learning information that can be represented with one's current conceptual repertoire; and conceptual construction, which involves acquiring knowledge that can only be represented in terms of concepts one does not yet possess. Theory changes involving conceptual change require conceptual construction. The cognitive mechanisms underlying conceptual change are still poorly understood, though executive function capacities have been implicated. The present study concerns the domain-general resources drawn upon in one well-studied case of the construction of a new framework theory in early childhood: the framework theory of vitalist biology, the ontogenetically earliest theory in which the concepts life and death come to have biological content shared with adults. Eighty-three five- and six-year-old children were tested on a battery of tasks that probe central concepts of the vitalist theory, as well as on a battery of tests of domain-general capacities that may be implicated in development in this domain, including measures of knowledge enrichment, executive function, and fluid IQ. With variance in accumulated knowledge and in knowledge enrichment capacity controlled, two specific executive functions, shifting and inhibition, predicted children's progress in constructing the vitalist theory. In contrast, working memory and fluid IQ were not associated with the acquisition of vitalist biology. These results provide further evidence for the distinction between knowledge enrichment and conceptual construction and impose new constraints on accounts of the mechanisms underlying conceptual construction in this domain.


A material with opioid activity in bovine milk and milk products.
Chloroform-methanol extracts of lyophilized milk, of commercially available dried milk or baby food and of casein digests were tested for opioid activity on the guinea-pig ileum longitudinal muscle-myenteric plexus preparation. Compounds with opioid activity--which proved to be resistant to peptidases--were detected in certain batches of baby food, casein digest, and cow milk in considerably varying amounts.


Probable presence of beta(1-2)-linked oligomannosides that act as human immunoglobulin G3 epitopes and are distributed over a Candida albicans 14- to 18-kilodalton antigen.
Kinetic analysis of candidosis patients' immunoglobulin G3 response has shown that reactivity towards beta(1-2)-linked mannan-derived oligomannosides was associated with the recognition through metaperiodate-sensitive epitopes of a 14- to 18-kDa Candida albicans antigen unreactive with concanavalin A.


Differential olfactory identification in children with autism and Asperger's disorder: a comparative and longitudinal study.
Key theories of autism implicate orbitofrontal cortex (OFC) compromise, while olfactory identification (OI) deficits are associated with OFC dysfunction. This study aimed to complete a 5-year follow-up of children with high-functioning autism (HFA) who previously lacked the normal age-OI association; and compare unirhinal-OI in children with HFA, Asperger's disorder (ASP), and controls. While both HFA and controls had improved birhinal-OI at follow-up, reduced OI in some HFA participants suggested OFC deterioration and heterogeneous OFC development. Unirhinal-OI was impaired in HFA but not ASP relative to controls, suggesting orbitofrontal compromise in HFA but integrity in ASP. Differing IQ-OI relationships existed between HFA and ASP. Findings support the hypothesis of separate neurobiological underpinnings in ASP and HFA, specifically differential orbitofrontal functioning.


A comparison of thermal enhancement of cis-diamminedichloroplatinum (II) induced renal and intestinal toxicities by whole body hyperthermia in the rat.
Thermal enhancement of cis-diamminedichloroplatinum (II) (DDP) induced renal and intestinal toxicities by whole body hyperthermia (WBH) were compared using a F344 rat model. Thermal enhancement ratios (TER) for DDP-induced nephrotoxicity were calculated using renal functional assays and morphological techniques. TER values for gastrointestinal (G.I.) toxicity were calculated using "severity of diarrhea" and jejunal crypt cell survival as assays. TER's for renal damage varied between 3 and 3.4, whereas the TER measured for G.I.-toxicity was 1.8. Physiological changes caused by WBH or intrinsic differences in the sensitivities of normal tissues to DDP +/- WBH may be responsible for the differences in thermal enhancement of DDP-induced renal and intestinal toxicities.