Chelation of cadmium without increased renal cadmium deposition.
Cadmium (Cd) is mainly accumulated in liver and kidney bound to metallothionein (MT) and excreted very slowly from the body. In chronic exposure, Cd is gradually transported from liver to kidney; the renal toxic effects appear when renal Cd concentration exceeds the critical concentration. In order to prevent the Cd-induced renal disease, it is important to control the movement of Cd to the kidney and its renal deposition. However, the chelation of Cd from liver is difficult because of the high affinity of intracellular MT for Cd. A number of chelating agents containing both carboxyl and thiol groups were able to mobilize and excrete Cd more easily in a short time (1/2 hr) after Cd exposure than longer times (24 hr), after MT synthesis. The renal deposition of Cd increased on BAL (2,3-dimercaptopropanol) treatment a short time (1/2 hr) after Cd exposure. However, it was observed that if BAL was administered 24 hr after Cd exposure, it could mobilize Cd from hepatic MT and increase the biliary excretion of Cd without any increase in renal Cd concentration. Studies using a number of structurally related thiols (mono-, di- and trithiols) showed that the major structural requirement for in vivo chelation of Cd from intracellular MT were the vicinal thiol groups on an aliphatic chain, and lipophilicity. BAL was the most effective of all the compounds studied and it did not mobilize Cd to the kidney, when most of the intracellular Cd was bound to MT. Furthermore, a delayed treatment with BAL or DTPA (diethylenetriamine pentaacetic acid) after synthesis of MT resulted in an increase in fecal or urinary excretion of Cd in rat model experiment.(ABSTRACT TRUNCATED AT 250 WORDS)


Transverse diffusion of laminar flow profiles to produce capillary nanoreactors.
We introduce transverse diffusion of laminar flow profiles (TDLFP), the first generic method for mixing two or more reactants inside capillaries. Conceptually, solutions of reactants are injected inside the capillary by pressure as a series of consecutive plugs. Due to the laminar nature of flow inside the capillary, the nondiffused plugs have parabolic profiles with predominantly longitudinal interfaces between them. After injection, the plugs are mixed by transverse diffusion; longitudinal diffusion does not contribute to mixing. To prove the principle, we used TDLFP to mix two reactants-an enzyme and its substrate. After mixing the reactants by TDLFP, we incubated reaction mixtures for different periods of time and measured the reaction kinetics. We found that the reaction proceeded in time- and concentration-dependent fashion, thus confirming that the reactants were mixed by TDLFP. Remarkably, the experimental reaction kinetics were not only in qualitative agreement but also in good quantitative agreement with theoretically predicted ones. TDLFP has a number of enabling features. By facilitating the preparation of reaction mixtures inside the capillary, TDLFP lowers reagent consumption to nanoliters (microliters are required for conventionally mixing reagents in a vial). The reaction products can be then analyzed "on-line" by capillary separation coupled with optical, electrochemical, or mass spectrometric detection. The combination of TDLFP with capillary separation will be an indispensable tool in screening large combinatorial libraries for affinity probes and drug candidates: a few microliters of a target protein will be sufficient to screen thousands of compounds. The new method paves the road to a wide use of capillary nanoreactors in different areas of physical and life sciences.


Ureases in the gastrointestinal tracts of ruminant and monogastric animals and their implication in urea-N/ammonia metabolism: A review.
Urea in diets of ruminants has been investigated to substitute expensive animal and vegetable protein sources for more than a century, and has been widely incorporated in diets of ruminants for many years. Urea is also recycled to the fermentative parts of the gastrointestinal (GI) tracts through saliva or direct secretory flux from blood depending upon the dietary situations. Within the GI tracts, urea is hydrolyzed to ammonia by urease enzymes produced by GI microorganisms and subsequent ammonia utilization serves the synthesis of microbial protein. In ruminants, excessive urease activity in the rumen may lead to urea/ammonia toxicity when high amounts of urea are fed to animals; and in non-ruminants, ammonia concentrations in the GI content and milieu may cause damage to the GI mucosa, resulting in impaired nutrient absorption, futile energy and protein spillage and decreased growth performance. Relatively little attention has been directed to this area by researchers. Therefore, the present review intends to discuss current knowledge in ureolytic bacterial populations, urease activities and factors affecting them, urea metabolism by microorganisms, and the application of inhibitors of urease activity in livestock animals. The information related to the ureolytic bacteria and urease activity could be useful for improving protein utilization efficiency in ruminants and for the reduction of the ammonia concentration in GI tracts of monogastric animals. Application of recent molecular methods can be expected to provide rationales for improved strategies to modulate urease and urea dynamics in the GI tract. This would lead to improved GI health, production performance and environmental compatibility of livestock production.


Sympathetic activity regulates epithelial proliferation and wound healing via adrenergic receptor α2A.
Innervation of the intestinal mucosa by the sympathetic nervous system is well described but the effects of adrenergic receptor stimulation on the intestinal epithelium remain equivocal. We therefore investigated the effect of sympathetic neuronal activation on intestinal cells in mouse models and organoid cultures, to identify the molecular routes involved. Using publicly available single-cell RNA sequencing datasets we show that the α2A isoform is the most abundant adrenergic receptor in small intestinal epithelial cells. Stimulation of this receptor with norepinephrine or a synthetic specific α2A receptor agonist promotes epithelial proliferation and stem cell function, while reducing differentiation in vivo and in intestinal organoids. In an anastomotic healing mouse model, adrenergic receptor α2A stimulation resulted in improved anastomotic healing, while surgical sympathectomy augmented anastomotic leak. Furthermore, stimulation of this receptor led to profound changes in the microbial composition, likely because of altered epithelial antimicrobial peptide secretion. Thus, we established that adrenergic receptor α2A is the molecular delegate of intestinal epithelial sympathetic activity controlling epithelial proliferation, differentiation, and host defense. Therefore, this receptor could serve as a newly identified molecular target to improve mucosal healing in intestinal inflammation and wounding.


Analysis of hot D2O emission using spectroscopically determined potentials.
Fourier transform emission spectra of D2O vapor were recorded at a temperature of 1500 degrees C in the wavenumber range 380-1880 cm(-1). 15 346 lines were measured, of which the majority were identified as belonging to D2O. The spectrum was analyzed using variational nuclear motion calculations based on spectroscopically determined potential-energy surfaces. Initial assignments were made using a potential surface obtained by fitting a high accuracy ab initio potential. The new assignments were used to refine the potential surface, resulting in additional assignments. A total of 6400 D2O transitions were assigned and 2144 new D2O energy levels were obtained. Transitions involving the 4nu2 and 5nu2 bending states, with band origins of 4589.30 (+/-0.02) and 5679.6 (+/-0.1) cm(-1), respectively, were assigned for the first time.


Treatment of Chronic Hepatitis C Can Improve Glycemic Control in Patients with Type 2 Diabetes.
A 73-year-old female patient with a history of type 2 DM for seven years was admitted to our out-patient clinic with a complaint of frequent hypoglycemic episodes. She was receiving basal- bolus insulin treatment. She underwent liver transplantation 20 months ago due to end stage liver disease caused by HCV infection genotype 1b. While she was still on tacrolimus for liver transplantation, she received direct acting antiviral agents including fix dose ledipasvir-sofosbuvir with ribavirin. Biochemical analysis showed fasting plasma glucose of 105 mg/dl and postprandial glucose of 200 mg/dl, glycosylated hemoglobin A1c of 4.8%, and c-peptide of 3.17 ng/ml. After achieving successfully virologic response with antiviral therapy, the patient stayed euglycemic and was no longer in need to any medication including insulin and the patient was followed only by dietary regulation. Achievement of the virological response in treatment of HCV infection can improve not only the liver status, but also the extrahepatic manifestations including type 2 DM.


Anderson localized state as a predissipative state: irreversible emission of thermalized quanta from a dynamically delocalized state.
It was shown that localization in one-dimensional disordered (quantum) electronic system is destroyed against coherent harmonic perturbations and the delocalized electron exhibits an unlimited diffusive motion [Yamada and Ikeda, Phys. Rev. E 59, 5214 (1999)]. The appearance of diffusion implies that the system has potential for irreversibility and dissipation. In the present paper, we investigate dissipative property of the dynamically delocalized state, and we show that an irreversible quasistationary energy flow indeed appears in the form of a "heat" flow when we couple the system with another dynamical degree of freedom. In the concrete we numerically investigate dissipative properties of a one-dimensional tight-binding electronic system perturbed by time-dependent harmonic forces, by coupling it with a quantum harmonic oscillator or a quantum anharmonic oscillator. It is demonstrated that if the on-site potential is spatially irregular an irreversible energy transfer from the scattered electron to the test oscillator occurs. Moreover, the test oscillator promptly approaches a thermalized state characterized by a well-defined time-dependent temperature. On the contrary, such a relaxation process cannot be observed at all for periodic potential systems. Our system is one of the minimal quantum systems in which a distinct nonequilibrium statistical behavior is self-induced.


Engineering zinc finger protein transcription factors to downregulate the epithelial glycoprotein-2 promoter as a novel anti-cancer treatment.
Zinc finger protein transcription factors (ZFP-TFs) are emerging as powerful novel tools for the treatment of many different diseases. ZFPs are DNA-binding motifs and consist of modular zinc finger domains. Each domain can be engineered to recognize a specific DNA triplet, and stitching six domains together results in the recognition of a gene-specific sequence. Inhibition of gene expression can be achieved by fusing a repressor domain to these DNA-binding motifs. In this study, we engineered ZFP-TFs to downregulate the activity of the epithelial glycoprotein-2 (EGP-2) promoter. The protein EGP-2 is overexpressed in a wide variety of cancer types and EGP-2 downregulation has been shown to result in a decreased oncogenic potential of tumor cells. Therefore, downregulation of EGP-2 expression by ZFP-TFs provides a novel anti-cancer therapeutic. Using a straightforward strategy, we engineered a 3-ZFP that could bind a 9 bp sequence within the EGP-2 promoter. After the addition of a repressor domain, this 3-ZFP-TF could efficiently downregulate EGP-2 promoter activity by 60%. To demonstrate the flexibility of this technology, we coupled an activation domain to the engineered ZFP, resulting in a nearly 200% increase in EGP-2 promoter activity. To inhibit the endogenous EGP-2 promoter, we engineered 6-ZFP-TFs. Although none of the constructed ZFP-TFs could convincingly modulate the endogenous promoter, efficient and specific inhibition of the exogenous promoter was observed. Overall, ZFP-TFs are versatile bi-directional modulators of gene expression and downregulation of EGP-2 promoter activity using ZFP-TFs can ultimately result in a novel anti-cancer treatment.


Cytotoxic aspidosperma-type alkaloids from Melodinus axillaris.
One new aspidosperma-type alkaloid, melotenine A (1), together with five known ones (2-6), was isolated from the leaves of Melodinus axillaris. Their structures were elucidated by detailed spectroscopic analysis and quantum chemical ECD calculation. The isolated aspidosperma-type alkaloids were evaluated for their cytotoxicity against four human cancer cell lines and melotenine A showed significant cytotoxicity against all cells with IC50 values ranging from 0.6 to 1.5 μM.


Urapidil analogues are potent ligands of the 5-HT1A receptor.
Urapidil and three derivatives with hypotensive properties (5-acetyl-, 5-formyl-, 5-methylurapidil) bind selectively to 5-HT receptors of the 5-HT1A subtype and to alpha 1-adrenoceptors labeled by [3H]8-OH-DPAT and [3H]prazosin, respectively. Binding to these receptors is likely to contribute to their hypotensive action. 5-Methylurapidil, the most potent of these drugs, was used in its 3H-labeled form as a radioligand. After blockade of alpha 1-adrenoceptors by prazosin, [3H]5-methylurapidil binds with nanomolar affinity to a binding site that is similar to the (5-HT1A) site labeled by [3H]8-OH-DPAT. No binding to other 5-HT1 and 5-HT2 receptors was observed. 5-HT uptake inhibitors did not inhibit [3H]5-methylurapidil binding. [3H]5-methylurapidil binding is sensitive to GTP and is modulated by divalent cations. Our results show that urapidil derivatives bind to the 5-HT1A recognition site and that [3H]5-methylurapidil is a valuable tool for the investigation of this receptor subtype.


High-speed broadband FTIR system using MEMS.
Current Fourier transform infrared spectroscopy (FTIR) systems have very good spectral resolution, but are bulky, sensitive to vibrations, and slow. We developed a new FTIR system using a microelectromechanical system (MEMS)-based lamellar grating interferometer that is fast, compact, and achromatic (i.e., does not require a beam splitter). The MEMS device has >10 mm2 active surface area, up to ±325 μm mechanical displacement, and a 343 Hz resonant operation frequency. The system uses a 5 MHz bandwidth custom infrared (IR) detector and a small emission area custom blackbody source to achieve fast interferogram acquisition and compact form factor. Effects of lamellar grating period, detector size, laser reference, apodization, and averaging of data on the spectral resolution are discussed. The measurement time ranges from 1.5 to 100 ms depending on the averaging time. In the target range of 2.5-16 μm (625-4000 cm-1) a spectral resolution of 15-20 cm-1 is demonstrated. The measurements are shown to be stable over a long time.


Design, Synthesis, and Biological Evaluation of Tetrahydro-α-carbolines as Akt1 Inhibitors That Inhibit Colorectal Cancer Cell Proliferation.
A series of densely functionalized THαCs were designed and synthesized as Akt1 inhibitors. Organocatalytic [3+3] annulation between indolin-2-imines 1 and nitroallylic acetates 2 provided rapid access to this pharmacologically interesting framework. In vitro kinase inhibitory abilities and cytotoxicity assays revealed that compound 3 af [(3S*,4S*)-4-(4-bromo-2-fluorophenyl)-9-methyl-3-nitro-1-tosyl-2,3,4,9-tetrahydro-1H-pyrido[2,3-b]indole] was the most potent Akt1 inhibitor, and mechanistic study indicated that compound 3 af suppressed the proliferation of colorectal cancer cells via inducing apoptosis and autophagy. Molecular docking suggested that the indole fragment of 3 af was inserted into the hydrophobic pocket of Akt1 protein, and the H-bond between 3 af and residue Lys179 also contributed to the stable binding. This article provides an efficient strategy to design and synthesize biologically important compounds as novel Akt1 inhibitors.


Lectins in the vulva. II. Vulvar intraepithelial neoplasia and squamous cell carcinoma.
This study used lectins as histologic probes to determine the cell surface oligosaccharide expression in different grades and types of vulvar intraepithelial neoplasia (VIN). Lectin binding patterns in metastasizing and non-metastasizing squamous cell carcinomas (SCCs) of the vulva were also compared to correlate lectin binding patterns with metastatic potential and other clinical/tumor characteristics. Twenty cases each of VIN epithelium, metastasizing SCC, and non-metastasizing vulvar carcinoma were randomly chosen from the pathology archives. Sixteen lectins were used to probe individual terminal oligosaccharide residues in formalin-fixed, paraffin-embedded tissue specimens from these cases through an indirect immunohistochemical technique. There were no differences in lectin binding patterns between the different histologic subtypes of VIN. In addition, there were no consistent differences between metastasizing and non-metastasizing primary tumors and no major differences in staining patterns between nodal metastases and the corresponding primary tumors. Furthermore, there was no identifiable correlation between lectin binding patterns and subsequent survival or local or regional recurrence; however, lectin staining of invasive tumor cells did appear to be related to local invasiveness. In addition, positive PNA binding was found to be a constant finding in each of the VIN and invasive SCC cases, confirming that the T-antigen becomes unmasked during the process of vulvar carcinogenesis. However, poorly-differentiated areas consistently showed absent lectin binding, suggesting loss of specific glycosyl transferase activities. In addition, the blood group "A" antigen appears to be lost during the process of tumorgenesis, although the blood group "O" antigen appears to be preserved.


Circulating soluble programmed death-1 levels may differentiate immune-tolerant phase from other phases and hepatocellular carcinoma from other clinical diseases in chronic hepatitis B virus infection.
Programmed death-1 (PD-1) is involved in the immune dysfunction of hepatitis B virus (HBV) infection and hepatocellular carcinoma (HCC). This study analyzed the association of circulating soluble PD-1 (sPD-1) levels with the phases and clinical diseases in chronic HBV infection. Serum sPD-1 levels were determined by enzyme linked immunosorbent assay in patients with different phases and liver diseases of chronic HBV infection. The sPD-1 levels in patients with chronic HBV infection were significantly elevated compared with HBV infection resolvers or healthy controls. According to phases, sPD-1 level in immune-tolerant phase (IT) was significantly lower than in other phases. Multivariate analysis showed that sPD-1 was an independent factor associated with IT. Area under the receiver operating characteristic (ROC) curves (AUC) showed that sPD-1 was significantly discriminative of IT from other phases with a cut-off of 1.535 ng/mL (AUC, 0.984; P<0.001). According to clinical diseases, sPD-1 level in HBV-related HCC was significantly higher than in other clinical diseases. Multivariate analysis showed that sPD-1 was an independent factor associated with HCC. The sPD-1 was significantly discriminative of HCC from other clinical diseases with a cut-off of 6.058 ng/mL (AUC, 0.962; P<0.001). The sPD-1 levels were significantly associated with HCC patients' overall survival. HCC resection resulted in remarkable reduction in sPD-1 levels. These results demonstrate the involvement of sPD-1 in the disease course of chronic HBV infection and indicate the potential to apply sPD-1 as a biomarker for differentiating IT from other phases and HCC from other disease conditions in chronic HBV infection.


Brushes of flexible, semiflexible, and rodlike diblock polyampholytes: Molecular dynamics simulation and scaling analysis.
Planar brushes of flexible, semiflexible, and rodlike diblock polyampholytes are studied using molecular dynamics simulations in a wide range of the grafting density. Simulations show linear dependence of the average thickness on the grafting density in all cases regardless of different flexibility of anchored chains and the brushes' different equilibrium conformations. Slopes of fitted lines to the average thickness of the brushes of semiflexible and rodlike polyampholytes versus the grafting density are approximately the same and differ considerably from that of the brushes of flexible chains. The average thickness of the brush of flexible diblock polyampholytes as a function of the grafting density is also obtained using a simple scaling analysis, which is in good agreement with our simulations.


Intraguild predation of Geocoris punctipes on Eretmocerus eremicus and its influence on the control of the whitefly Trialeurodes vaporariorum.
Geocoris punctipes (Hemiptera: Lygaeidae) and Eretmocerus eremicus (Hymenoptera: Aphelinidae) are whitefly natural enemies. Previously, under laboratory conditions, we showed that G. punctipes engages in intraguild predation (IGP), the attack of one natural enemy by another, on E. eremicus. However, it is unknown whether this IGP interaction takes place under more complex scenarios, such as semi-field conditions. Even more importantly, the effect of this interaction on the density of the prey population requires investigation. Therefore, the present study aimed to establish whether this IGP takes place under semi-field conditions and to determine whether the predation rate of G. punctipes on the whitefly decreases when IGP takes place. Molecular analysis showed that, under semi-field conditions, G. punctipes performed IGP on E. eremicus. However, although IGP did take place, the predation rate by G. punctipes on the whitefly was nevertheless higher when both natural enemies were present together than when the predator was present alone. While IGP of G. punctipes on E. eremicus does occur under semi-field conditions, it does not adversely affect whitefly control. The concomitant use of these two natural enemies seems a valid option for inundative biological control programmes of T. vaporariorum in tomato. © 2015 Society of Chemical Industry.


The formation of calcium lactate crystals is responsible for concentrated acid whey thickening.
The use of spray drying for dehydration of acid whey is generally limited by the appearance of uncontrolled thickening and solidifying of the whey mass during the lactose crystallization step. The origin of this physical change is still unknown and probably linked to complex interactions between physical properties and chemical composition of these products. To understand this phenomenon, we simulated the thickening of concentrated acid whey on a laboratory scale by measuring the flow resistance changes as a function of time and whey composition. The thickening process was characterized by an amplitude of torque and a lag time (induction time). Thickening of lactic acid whey concentrate occurred regardless of the presence of whey proteins or lactose crystals. Moreover, this work clearly demonstrated that the thickening process was due to the formation of filamentous structures corresponding to calcium lactate crystals and showed a large dependence on calcium and lactate contents, pH, and phosphate concentration.


[Thalamic neuro inhibition in the treatment of post traumatic tremor].
Severe head trauma could be followed by a wide range of movement disorders. Usually, they appear as a moderate or severe intention tremor that causes great patient disability. Those movement disorders use to be refractary to medical treatment, although sometimes they would be spontaneusly controlled by a year. 14 year old boy who suffered severe head trauma at age of 8. One year after the injury he developed an action tremor and choreiform dyskinesia with hemibalistic features evocated by the movement. Medical treatment was ineffective. Due to the great patients disability, we chose the surgical treatment based on the placement of an stimulation electrode in thalamic Vop/Vim complex, obtaining a tremor abolition with an improvement in the scales of tremor and quality of life. Surgical treatment of postraumatic tremors tryes to improve in terms of function and activities of daily living, more than suppresion of a symptom. The aim of this treatment is cut the palidal thalamic cortical pathways. Anyway, target is defined by the tremor related cells and, it is more a functional target than an anatomical one. We prefer stimulation more than lesion, due to the lack of brain definitive injury because its reversibility and adjustability features.


MYD88 L265P is a marker highly characteristic of, but not restricted to, Waldenström's macroglobulinemia.
We evaluated the MYD88 L265P mutation in Waldenström's macroglobulinemia (WM) and B-cell lymphoproliferative disorders by specific polymerase chain reaction (PCR) (sensitivity ∼10(-3)). No mutation was seen in normal donors, while it was present in 101/117 (86%) WM patients, 27/31 (87%) IgM monoclonal gammapathies of uncertain significance (MGUS), 3/14 (21%) splenic marginal zone lymphomas and 9/48 (19%) non-germinal center (GC) diffuse large B-cell lymphomas (DLBCLs). The mutation was absent in all 28 GC-DLBCLs, 13 DLBCLs not subclassified, 35 hairy cell leukemias, 39 chronic lymphocytic leukemias (16 with M-component), 25 IgA or IgG-MGUS, 24 multiple myeloma (3 with an IgM isotype), 6 amyloidosis, 9 lymphoplasmacytic lymphomas and 1 IgM-related neuropathy. Among WM and IgM-MGUS, MYD88 L265P mutation was associated with some differences in clinical and biological characteristics, although usually minor; wild-type MYD88 cases had smaller M-component (1.77 vs 2.72 g/dl, P=0.022), more lymphocytosis (24 vs 5%, P=0.006), higher lactate dehydrogenase level (371 vs 265 UI/L, P=0.002), atypical immunophenotype (CD23-CD27+ +FMC7+ +), less Immunoglobulin Heavy Chain Variable gene (IGHV) somatic hypermutation (57 vs 97%, P=0.012) and less IGHV3-23 gene selection (9 vs 27%, P=0.014). These small differences did not lead to different time to first therapy, response to treatment or progression-free or overall survival.


[Economic glossary: pitfalls of interpretation].
Sharp discrepancy exists between the concepts declared and the measures introduced with urgency by the government and the real praxis of the health-care institutions, which are delivered to and suffer from the heaviest financial-economical restrictions. The situation may partly be attributed to the missing system-specific interpretation of some leading categories. Attempts were made to point at some harmful consequences of this failure and to reinterpret basic categories, so as to contribute to a better agreement of the participants in favour of our patients' welfare.


[Pediatric palliative care].
Based on the definition by the WHO from 1998, palliative care for children is the active, total care of the childs body, mind and spirit, and involves giving care to the family. It begins when the illness is diagnosed and continues regardless of whether a child receives treatment directed at the disease. The World Health Organization (WHO) definition of pediatric palliative care was adopted by IMPaCCT (International Meeting for Palliative Care in Children, Trento) and became the key point of document, which defines and determines standards for care of children with life-limiting and life-threatening disease and recommends that these standards should be implemented in all European countries. Since 2015, the development of child palliative care in the Czech Republic is systematically promoted by the task force on palliative care for children of the Czech Society of Palliative Medicine.Key words: pediatric palliative care, life-limited disease, life-threatening disease, standards, family, complex care.


Rad GTPase is essential for the regulation of bone density and bone marrow adipose tissue in mice.
The small GTP-binding protein Rad (RRAD, Ras associated with diabetes) is the founding member of the RGK (Rad, Rem, Rem2, and Gem/Kir) family that regulates cardiac voltage-gated Ca2+ channel function. However, its cellular and physiological functions outside of the heart remain to be elucidated. Here we report that Rad GTPase function is required for normal bone homeostasis in mice, as Rad deletion results in significantly lower bone mass and higher bone marrow adipose tissue (BMAT) levels. Dynamic histomorphometry in vivo and primary calvarial osteoblast assays in vitro demonstrate that bone formation and osteoblast mineralization rates are depressed, while in vitro osteoclast differentiation is increased, in the absence of Rad. Microarray analysis revealed that canonical osteogenic gene expression (Runx2, osterix, etc.) is not altered in Rad-/- calvarial osteoblasts; instead robust up-regulation of matrix Gla protein (MGP, +11-fold), an inhibitor of extracellular matrix mineralization and a protein secreted during adipocyte differentiation, was observed. Strikingly, Rad deficiency also resulted in significantly higher marrow adipose tissue levels in vivo and promoted spontaneous in vitro adipogenesis of primary calvarial osteoblasts. Adipogenic differentiation of wildtype calvarial osteoblasts resulted in the loss of endogenous Rad protein, further supporting a role for Rad in the control of BMAT levels. These findings reveal a novel in vivo function for Rad and establish a role for Rad signaling in the complex physiological control of skeletal homeostasis and bone marrow adiposity.


Observed differences in gentamicin pharmacokinetic parameters and dosage recommendations determined by fluorescent polarization immunoassay and radioimmunoassay methods.
Radioimmunoassay (RIA) and fluorescent polarization immunoassay (FPI) methods for quantitative gentamicin serum concentration assay have been shown to be comparable. The purpose of this study was to determine if serum concentration-time data from the same patients assayed by RIA and FPI would provide the same estimates for half-life, elimination rate constant, distribution volume, drug clearance, and gentamicin dose. A total of 99 pre-and postinfusion serum samples were obtained from 30 patients. Samples were divided and assayed by RIA or FPI, and the resultant serum concentration-time data were fitted to a standard one-compartment model. The correlation between the two assay methods was 0.99 (p less than 0.005). A mean difference of 10% was seen in distribution volume, gentamicin clearance, and gentamicin dose from quantitative data from the two methods. These differences were significant (p less than 0.01). Although the two methods appear to be interchangeable, based on in vitro comparison, differences in calculated pharmacokinetic parameters resulted in significant differences in dose recommendations.


Gender inequalities in mental wellbeing in 26 European countries: do welfare regimes matter?
Nature and extent of welfare regimes and social policies are important determinants of health and health inequalities. This study examines the association of gender and mental wellbeing in European countries and investigates whether type of welfare regime plays a role in this association. Data of 19 366 women and 14 338 men of the third round of the European Quality of Life Survey (2011-12) was used to analyse mental wellbeing, assessed by the World Health Organization 5-Mental Wellbeing Index. Multilevel logistic regression analyses were performed to analyse the association between gender and good mental wellbeing first at country-level, and secondly the between country variation was analysed and welfare regimes were included as explanatory variables. We observed cross-national variation in good mental wellbeing. At country levels gender inequalities in good mental wellbeing were observed in 7 out of 26 countries. In analyses considering all countries together gender inequalities in good mental wellbeing were identified independent of further individual socio-demographic variables and independent of the welfare regimes that people lived in [women vs. men: OR = 0.76; (95% CI = 0.71-0.81)]. Gender inequalities in good mental wellbeing were not modified by welfare regimes. There are cross-national differences in good mental wellbeing between European countries. Gender inequalities with a lower prevalence of good mental wellbeing among women are common in European countries. This study suggests that welfare regimes do not modify these gender inequalities in mental wellbeing.


Human umbilical cord matrix stem cells: preliminary characterization and effect of transplantation in a rodent model of Parkinson's disease.
The umbilical cord contains an inexhaustible, noncontroversial source of stem cells for therapy. In the U.S., stem cells found in the umbilical cord are routinely placed into bio-hazardous waste after birth. Here, stem cells derived from human umbilical cord Wharton's Jelly, called umbilical cord matrix stem (UCMS) cells, are characterized. UCMS cells have several properties that make them of interest as a source of cells for therapeutic use. For example, they 1) can be isolated in large numbers, 2) are negative for CD34 and CD45, 3) grow robustly and can be frozen/thawed, 4) can be clonally expanded, and 5) can easily be engineered to express exogenous proteins. UCMS cells have genetic and surface markers of mesenchymal stem cells (positive for CD10, CD13, CD29, CD44, and CD90 and negative for CD14, CD33, CD56, CD31, CD34, CD45, and HLA-DR) and appear to be stable in terms of their surface marker expression in early passage (passages 4-8). Unlike traditional mesenchymal stem cells derived from adult bone marrow stromal cells, small populations of UCMS cells express endoglin (SH2, CD105) and CD49e at passage 8. UCMS cells express growth factors and angiogenic factors, suggesting that they may be used to treat neurodegenerative disease. To test the therapeutic value of UCMS cells, undifferentiated human UCMS cells were transplanted into the brains of hemiparkinsonian rats that were not immune-suppressed. UCMS cells ameliorated apomorphine-induced rotations in the pilot test. UCMS cells transplanted into normal rats did not produce brain tumors, rotational behavior, or a frank host immune rejection response. In summary, the umbilical cord matrix appears to be a rich, noncontroversial, and inexhaustible source of primitive mesenchymal stem cells.


[Development of reversible oviductal polyethylene contraceptive plugs].
An introduction of the design, features and contraceptive principle of an oviduct contraceptive plug is given in this paper. The plug is made of polyethylene, based on the oviductal morphology. It can be introduced and removed by a hysteroscope and is expected to be a transcervical, implantable and permanent intratubal contraceptive device.


32P-postlabeling of bile components: bulky adduct-like behavior in polyethyleneimine-cellulose thin layer chromatography.
The 32P-postlabeling assay has been used widely in carcinogen-DNA adduct analysis because of its sensitivity and reproducibility. Cloned T4 polynucleotide kinase (PNK), routinely used in this assay, phosphorylates the 5'-OH groups of adducted nucleotides in the presence of [gamma-32P]ATP. However, as an exception to this property, PNK has been reported to phosphorylate non-adducted carcinogen metabolites, such as tetrol derivatives of benzo[a]pyrene and chrysene. Also, PNK phosphorylates both 5'-OH and 3'-OH groups of safrole-adducted deoxydinucleoside monophosphates having an unmodified purine in the 3'-position. In the present study we show that T4 PNK catalyzed the transfer of [32P]phosphate from [gamma-32P]ATP to rat bile components or purified bile acids (derivatives of 3 alpha-hydroxy-5 beta-cholanic acid) in the absence of nucleic acids or nucleases. However, labeling of the bile acids appeared over 100,000-fold less efficient than labeling of 2'-deoxyadenosine-5'-monophosphate. There was no reaction in the absence of bile components or PNK. Dehydrocholic acid, which lacks hydroxyl groups, was resistant to phosphorylation. On polyethyleneimine-cellulose TLC maps, 32P-labeled rat bile extract gave an array of non-polar radioactive spots which resembled carcinogen-DNA adducts, while 32P-labeled purified bile acids each gave a single spot. These 32P-labeled products liberated 32Pi upon incubation with prostatic acid phosphatases. Two of the radioactive spots obtained from rat bile were identified as phosphorylated taurocholic and taurodeoxycholic acids by co-chromatography with 32P-labeled standards. These findings demonstrate for the first time that PNK is able to phosphorylate natural products other than nucleotides and further emphasize the need to rule out contamination with bile acids and possibly other bulky/hydrophobic alcohols when analyzing DNA samples by 32P-postlabeling.


Abnormal Indexes of Liver and Kidney Injury Markers Predict Severity in COVID-19 Patients.
SARS-CoV-2 can damage not only the lungs but also the liver and kidney. Most critically ill patients with coronavirus disease 2019 (COVID-19) have liver and kidney dysfunction. We aim to investigate the levels of liver and kidney function indexes in mild and severe COVID-19 patients and their capability to predict the severity of the disease. The characteristics and laboratory indexes were compared between patients with different conditions. We applied binary logistic regression to find the independent risk factors of severe patients. Receiver operating characteristic (ROC) analysis was used to predict the severity of COVID-19 using the liver and kidney function indexes. This study enrolled 266 COVID-19 patients, including 235 mild patients and 31 severe patients. Compared with mild patients, severe patients had lower albumin (ALB) and higher alanine aminotransferase (ALT), aspartate aminotransferase (AST), and urea nitrogen (BUN) (all p<0.001). Binary logistic regression analysis also identified ALB [OR=0.273 (0.079-0.947), p=0.041] and ALT [OR=2.680 (1.036-6.934), p=0.042] as independent factors of severe COVID-19 patients. Combining ALB, ALT, BUN, and LDH exhibited the area under ROC at 0.914, with a sensitivity of 86.7% and specificity of 83.0%. COVID-19 patients, especially severe patients, have damage to liver and kidney function. ALT, AST, LDH, and BUN could be independent factors for predicting the severity of COVID-19. Combining the ALB, ALT, BUN, and LDH could predict the transition from mild to severe in COVID-19 patients.


Pteridine analysis in urine by capillary electrophoresis using laser-induced fluorescence detection.
Pteridines are a class of compounds excreted in urine, the levels of which are found to elevate significantly in tumor-related diseases. For the first time, we have developed a method, based on high-performance capillary electrophoresis (HPCE) and laser-induced fluorescence (LIF) detection, to monitor the pteridine levels in urine. HPCE provides better separation than high-performance liquid chromatography and the LIF detector enables us to detect minute amounts of pteridines in body fluid. Eight different pteridine derivatives were well separated in 0.1 M Tris-0.1 M borate-2 mM EDTA buffer (pH 8.75) using a 60-cm fused-silica capillary (50-micron i.d., 35-cm effective length), six of which were detected and characterized in urine samples from normal persons and different cancer patients. The detection limits of these pteridines are under 1 x 10(-10) M. The levels of neopterin, pterine, xanthopterin, and pterin-6-carboxylic acid were found to be significantly elevated in urine excreted by cancer patents, while the level of isoxanthopterin dropped in these patients. No significant change of biopterin level was found between healthy individuals and cancer patients. This method can be used in clinical laboratories either for cancer monitoring or for precancer screening.


Llyod berkner dies at 62.
In the obituary on Llyod V. Berkner (News and Comment, 9 June, p. 1349) two erroneous statements were made. Berkner was not ". . . principal administrator of the U. S. part of [IGY] program during its operation in 1957 and 1958" as stated in paragraph 4. In paragraph 9, it is stated that " He was also radio man on the first air flight over the South Pole." Berkner was a member of that expedition, but he was not on that flight. Harold June was the radio operator.


Evolution of the poliomyelitis vaccination program in Costa Rica.
Outbreaks of poliomyelitis in Costa Rica occurred at variable intervals until 1972. The first campaign with Salk vaccine was in 1956 and the first with Sabin vaccine was in 1959. Since then, campaigns have been irregular, depending on the occurrence of outbreaks. Continuous vaccination programs were inaugurated after the last outbreak in 1972, and the coverage of primary health care in rural areas was extended. These programs resulted in high levels of immunization. Since no cases of poliomyelitis have occurred in the last 10 years, we can say confidently that the disease has been eradicated in Costa Rica.


Rare case of Cushing's disease due to double ACTH-producing adenomas, one located in the pituitary gland and one into the stalk.
We describe a patient affected by Cushing's disease due to the presence of double pituitary adenomas, one located within the anterior pituitary and the other in the infundibulum associated with a remnant of Rakthe's pouch. Cure was achieved only after the infundibulum lesion was surgically removed. A 38-year-old female presented with unexplained weight gain, hirsutism, amenorrhea, asthenia, recurrent cutaneous micotic infections and alopecia. Hormonal studies indicated Cushing's disease and MRI showed an enlarged pituitary gland with a marked and homogeneous enhancement after injection of gadolinium and an enlarged infundibulum with a maximum diameter of 8 mm. As a venous sampling of the inferior petrosal sinus after 10 μg iv desmopressin stimulation revealed a central to peripheral ACTH ratio consistent with a pituitary ACTH-secreting tumor, transphenoidal explorative surgery was performed and a 4-mm pituitary adenoma immunopositive for ACTH was disclosed and removed. Since postoperative hormonal evaluation showed persistent hypercortisolism, confirmed by dynamic tests, the patient again underwent surgery by transcranial access and the infundibulum mass was removed. Histology and immunochemistry were consistent with an ACTH-secreting adenoma. A few months after the second operation, cushingoid features were significantly reverted and symptoms improved. Although Cushing's patients bearing multiple adenomas have already been documented, the presence of two adenomas both immunohistochemically positive for ACTH is a very rare cause of Cushing's disease and this is the first report of a case of double ACTH-producing adenomas, one located in the pituitary gland and one attached to the stalk.


A central thermogenic-like mechanism in feeding regulation: an interplay between arcuate nucleus T3 and UCP2.
The active thyroid hormone, triiodothyronine (T3), regulates mitochondrial uncoupling protein activity and related thermogenesis in peripheral tissues. Type 2 deiodinase (DII), an enzyme that catalyzes active thyroid hormone production, and mitochondrial uncoupling protein 2 (UCP2) are also present in the hypothalamic arcuate nucleus, where their interaction and physiological significance have not been explored. Here, we report that DII-producing glial cells are in direct apposition to neurons coexpressing neuropeptide Y (NPY), agouti-related protein (AgRP), and UCP2. Fasting increased DII activity and local thyroid hormone production in the arcuate nucleus in parallel with increased GDP-regulated UCP2-dependent mitochondrial uncoupling. Fasting-induced T3-mediated UCP2 activation resulted in mitochondrial proliferation in NPY/AgRP neurons, an event that was critical for increased excitability of these orexigenic neurons and consequent rebound feeding following food deprivation. These results reveal a physiological role for a thyroid-hormone-regulated mitochondrial uncoupling in hypothalamic neuronal networks.


Developments in development: What have we learned from primate locomotor ontogeny?
The importance of locomotion to evolutionary fitness has led to extensive study of primate locomotor behavior, morphology and ecology. Most previous research has focused on adult primates, but in the last few decades, increased attention to locomotor development has provided new insights toward our broader understanding of primate adaptation and evolution. Here, we review the contributions of this body of work from three basic perspectives. First, we assess possible determinants on the timing of locomotor independence, an important life history event. Significant influences on timing of locomotor independence include adult female body mass, age at weaning, and especially relative brain size, a significant predictor of other primate life history variables. Additionally, we found significant phylogenetic differences in the timing of locomotor independence, even accounting for these influences. Second, we discuss how structural aspects of primate growth may enhance the locomotor performance and safety of young primates, despite their inherent neuromotor and musculoskeletal limitations. For example, compared to adults, growing primates have greater muscle mechanical advantage, greater bone robusticity, and larger extremities with relatively long digits. Third, focusing on primate quadrupedalism, we provide examples that illustrate how ontogenetic transitions in morphology and locomotion can serve as a model system for testing broader principles underlying primate locomotor biomechanics. This approach has led to a better understanding of the key features that contribute to primates' stride characteristics, gait patterns, limb force distribution, and limb postures. We have learned a great deal from the study of locomotor ontogeny, but there is much left to explore. We conclude by offering guidelines for future research, both in the laboratory and the field.


Aberrations of the tumor suppressor p53 gene and p53 protein in solar keratosis in human skin.
Aberrations of the p53 gene in 27 solar keratoses were examined by the polymerase chain reaction and single-strand conformation polymorphism and DNA sequencing analyses. In a series of Japanese patients, eight of 27 cases (30%) of solar keratosis showed structural abnormalities of the p53 gene. Six of eight aberrations of p53 gene were determined to be single nucleotide substitutions, and five of these were located at a dipyrimidine site. In solar keratosis, noticeable mutations were C to T in three cases, and one each of C to A and T to C nucleotide changes. p53 protein was detected immunohistochemically in the nuclei of six of 27 cases (22%) of solar keratosis. Nuclear staining for p53 protein was only significantly correlated with the presence of missense mutation of p53 gene (p < 0.01). Aberrations of the p53 gene in solar keratosis may be a marker to predict early cancerous lesions.


[A woman with onychomatricoma of a toe nail].
The authors present a 43-year-old woman with a yellow, swollen toe nail of the right foot. This nail was lesioned 20 years before. The proximal nail fold of the right hallux was erythematous and indurated. The medial half of the nail was thickened and yellowish, sharply demarcated from the unaffected half. This aspect is typical for onychomatricoma.


Demyelinating Peripheral Neuropathy Caused by the p.R160H Mutation in the LITAF Gene.
We report a 62-year-old woman who presented with complaints of numbness and tingling in her feet without a family history suggestive of neuropathy. Neurological examination and electromyogram testing confirmed the presence of a demyelinating neuropathy with a mild phenotype. Extensive testing revealed no etiology and she was diagnosed and treated unsuccessfully for chronic inflammatory demyelinating polyneuropathy. Ultimately, with the availability of next-generation sequencing, genetic testing revealed a heterozygous variant, chr16:11643500C > T, c.479 G > A, p.R160H, in the lipopolysaccharide-induced tumor necrosis factor (LITAF ) gene. Further analysis of this variant employing protein modeling suggests that this is a disease producing mutation causing Charcot Marie Tooth disease type 1C (CMT1C). Our study demonstrates the power of next-generation sequencing to diagnose patients with idiopathic neuropathy. This is important as it avoids unnecessary and expensive treatments for the patient and furthermore, allows genetic counseling for family members.


Mucosa-Associated Microbial Profile Is Altered in Small Intestinal Bacterial Overgrowth.
The overall gut microbial profile of patients with small intestinal bacterial overgrowth (SIBO) has not been thoroughly investigated. We investigated the microbial communities of mucosal specimens from the duodenum, ileum, sigmoid colon, and feces of patients with and without SIBO, as diagnosed by lactulose breath testing. The bacteria present in the mucosal and fecal samples were identified using 16S rRNA gene sequencing. Further analysis was performed using the linear discriminant analysis (LDA) effect size method, random forest analysis, and receiver operating characteristic analysis. The microbial diversities of the fecal samples were significantly lower than those of the mucosal samples from the duodenum, ileum, and sigmoid colon (P < 0.001, P < 0.001, and P < 0.001, respectively), while the bacterial compositions of the ileac mucosal samples and sigmoid mucosal samples were similar. The bacterial composition of either the fecal or duodenal mucosal samples were significantly different from those of the other three groups (ANOSIM R = 0.305, P = 0.001). The bacterial compositions of the mucosal samples of the duodenum, ileum, and sigmoid colon in the SIBO + subjects were significantly different from those of the SIBO- subjects (ANOSIM P = 0.039, 0.002, and 0.007, respectively). The relative abundances of 7, 18, and 8 genera were significantly different (LDA score > 3) in the mucosal samples of the duodenum, ileum, and sigmoid colon between the SIBO + and SIBO- groups. Four genera (Lactobacillus, Prevotella_1, Dialister, and norank_f__Ruminococcaceae) showed similar changes among the mucosal samples of the duodenum, ileum, and sigmoid colon in the SIBO + subjects. A signature consisting of four genera in the duodenal mucosa, three genera in the ileac mucosa, or six genera in the mucosa of the sigmoid colon exhibited predictive power for SIBO (area under the curve = 0.9, 0.93, and 0.87, respectively). This study provides a comprehensive profile of the gut microbiota in patients with SIBO. Dysbiosis was observed in the mucosa-associated gut microbiome but not in the fecal microbiome of patients with SIBO. Furthermore, we identified mucosa-associated taxa that may be potential biomarkers or therapeutic targets of SIBO. Further investigation is needed on their mechanisms and roles in SIBO.


[Driver's license, driving habits and traffic safety of patients with diabetes mellitus].
Motoring practices of diabetic drivers are similar to those of the average driving population. Due to the sudden onset of disabling hypoglycemia and the long-term complications of the disease, particularly retinopathy, one might assume that diabetic drivers are more prone to road traffic accidents than the average driver; however, the risk is not increased. The Council of the European Union recently laid down guidelines for dealing with this problem, which will lead to the introduction of a new law in Austria governing driving licensing. This legislation is currently under consideration. We recommend that discrimination between patients requiring therapeutic regimens with potential hypoglycemic side effects (insulin and/or sulphonylureas) and those who are not at risk of hypoglycemia (controlled by diet only or oral antidiabetic drugs other than sulphonylureas) should be legally defined by the licensing authorities. Furthermore, the legal enforcement of participation by diabetic drivers in a structured teaching programme, with the special aim of informing on traffic attitudes and avoidance of hypoglycemia while driving, might be of great importance for safety on the road.


Tumor necrosis factor-alpha antagonists twenty years later: what do Cochrane reviews tell us?
In 1998, the U.S. Food and Drug Administration granted regulatory approval to the first tumor necrosis factor-α antagonist, infliximab, for the treatment of moderately to severely active Crohn's disease. As of 2013, there were 3 additional tumor necrosis factor-α antagonists commercially available for the treatment of inflammatory bowel disease in the United States: adalimumab, certolizumab pegol, and golimumab. Despite a vast literature describing both clinical trial and clinical practice experience with these agents, there remain important questions regarding the efficacy and safety of tumor necrosis factor-α antagonists for the treatment of inflammatory bowel disease. These questions and the best available evidence to answer them were discussed during a Cochrane Collaboration session held at the 2013 Digestive Diseases Week annual meeting. This article reviews the data from that session.


Stereotactic radiosurgery for malignant brain tumors: the patient's perspective.
Stereotactic radiosurgery is a relatively new approach used to treat primary malignant brain tumors. Radiosurgery delivers a single-fraction, high dose irradiation to a limited target volume of tissue. Stereotactic radiosurgery was conceived to be more analogous to conventional surgery than to conventional radiotherapy. Eight patients who had primary malignant brain tumors were purposefully sampled to participate in this phenomenology study in an effort to describe the experience and meaning of the treatment from the patient's perspective. Using Colaizzi's phenomenological technique, five potential themes emerged from the interviews obtained. They are; (1) A willingness of patients to undergo treatment again, (2) self-directed education regarding the treatment method, (3) an ability to verbalize the prognosis for their tumor type, (4) the lack of discomfort during treatment and (5) the fact that this treatment was not originally a treatment option. Additional studies need to be conducted with patients undergoing stereotactic radiosurgery for vascular and nonmalignant conditions. Implications for nursing practice include an understanding of what the experience means to patients. Teaching plans that focus on the patient's needs have been developed as a result of this study.


The control of ribosomal RNA transcription in lymphocytes. Evidence that the rate of chain elongation is the limiting factor.
During activation of lymphocytes by phytohaemagglutinin, the nuclear activity of RNA polymerase I increases with no proportional increase in the amount of catalytic efficiency of the enzyme in the cell. The mechanism by which rRNA transcription in lymphocytes is modified by phytohaemagglutinin stimulation was investigated. The following results were obtained. (a) In resting lymphocytes all RNA polymerase II molecules are bound to the template while a pool of excess free RNA polymerase I, not engaged in transcription, can be detected by its ability to transcribe added poly[d(A-T)]. (b) Although the free RNA polymerase I activity increases twofold to threefold during phytohaemagglutinin stimulation, there is no evidence that the free enzymes ever become engaged in transcription. (c) Most of the RNA chains in elongation in nuclei from resting lymphocytes are being elongated by class II RNA polymerase and their rate of elongation is much higher than that of other RNA species. (d) The same number of pre-rRNA chains are in the process of being elongated in nuclei from resting and stimulated lymphocytes. However, the rate of elongation of pre-rRNA, which is slow relative to the average in resting lymphocytes, increases twofold to threefold within 6 h of phytohaemagglutinin stimulation and rises to sixfold by 19 h. These results suggest that the control of rRNA transcription in phytohaemagglutinin-stimulated lymphocytes lies in the elongation step of transcription rather than in initiation, and that little or no additional rRNA template is transcribed in phytohaemagglutinin-stimulated lymphocytes.


Comparative proteomic analysis of indica and japonica rice varieties.
Indica and japonica are two main subspecies of Asian cultivated rice (Oryza sativa L.) that differ clearly in morphological and agronomic traits, in physiological and biochemical characteristics and in their genomic structure. However, the proteins and genes responsible for these differences remain poorly characterized. In this study, proteomic tools, including two-dimensional electrophoresis and mass spectrometry, were used to globally identify proteins that differed between two sequenced rice varieties (93-11 and Nipponbare). In all, 47 proteins that differed significantly between 93-11 and Nipponbare were identified using mass spectrometry and database searches. Interestingly, seven proteins were expressed only in Nipponbare and one protein was expressed specifically in 93-11; these differences were confirmed by quantitative real-time PCR and proteomic analysis of other indica and japonica rice varieties. This is the first report to successfully demonstrate differences in the protein composition of indica and japonica rice varieties and to identify candidate proteins and genes for future investigation of their roles in the differentiation of indica and japonica rice.


Identification of cell surface binding sites for PC-cell-derived growth factor, PCDGF, (epithelin/granulin precursor) on epithelial cells and fibroblasts.
PC cell derived growth factor (PCDGF) is an 88-kDa glycoprotein purified from the culture medium of the highly tumorigenic mouse teratoma-derived cell line PC. PCDGF was shown to stimulate the proliferation of 3T3 fibroblasts and PC cells. Amino acid sequencing of PCDGF indicated its identity to the precursor for the 6-kDa polypeptides epithelins and granulins. In this paper, we investigated the binding of PCDGF to the mink lung epithelial cell line CCL64. Scatchard analysis indicates that 125I-PCDGF binding to CCL64 cells is curvilinear, corresponding to the existence of two classes of binding sites: high affinity binding sites (560 +/- 170 sites/cell) with a Kd1 of 43 +/- 15 pM and low affinity binding sites (16,350 +/- 5900 sites/cell) with a Kd2 of 3.9 +/- 1.9 nM. 125I-PCDGF was chemically crosslinked to cell surface receptors on CCL64 cells with disuccinimidyl suberate. A major crosslinked band of about 190 kDa with radiolabeled PCDGF was detected after SDS-PAGE, suggesting the presence of PCDGF binding sites with molecular weight of about 120 kDa. 125I. PCDGF crosslinking studies indicate the presence of PCDGF binding sites with a molecular weight similar to those of binding sites on CCL64 cells on the surface of two other PCDGF-responsive cell lines, 3T3 fibroblasts and PC cells. These data suggest that the receptors for PCDGF are widely distributed on cells of distinct embryonic origin.


Apoptosis mediated by lentiviral TRAIL transfer involves transduction-dependent and -independent effects.
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anticancer agent, which selectively induces apoptosis in many transformed cells without apparent toxic side effects in normal tissue. We recently described the construction and characterization of a lentiviral vector for expression of TRAIL. In this report, we evaluate its suitability for therapeutic application. In vitro, we observed specific induction of apoptosis upon transduction in human lung cancer cells. Cell death was partially dependent on successful integration and TRAIL expression by the vectors, but was to some extent mediated by protein carryover, as we found TRAIL protein associated with virus particles. Transduction of subcutaneously growing lung tumors on nude mice with lentiviral TRAIL mediated a transient suppression of tumor growth. Analysis of tumor sections revealed that transduction efficiency of lentiviral control vector but not of lentiviral TRAIL vector was high. This was because of the direct cytotoxic activity of recombinant TRAIL present in viral particles, which prevented efficient tumor transduction. These data therefore suggest that enveloped viral vectors constitutively expressing TRAIL are well suited for ex vivo applications, such as the transduction of tumor-homing cells, but may have a lower effect when used directly for the transduction of tumor cells in vivo.


Blood, brain, and cerebrospinal fluid concentrations of several antibiotics in rabbits with intact and inflamed meninges.
Because cerebrospinal fluid (CSF) antibiotic levels fail to predict either clinical success or relapse in the treatment of bacterial meningitis, we examined simultaneous antibiotic concentrations in the blood, brain, and CSF of control rabbits and of animals with experimental pneumococcal meningitis. Cefamandole pharmacokinetics were analyzed in detail and compared with those of cephalothin, ampicillin, penicillin G, and tobramycin. After 4 h of continuous intravenous infusion, cefamandole reached concentrations in both brain and CSF in excess of the minimal bactericidal concentration for the test organism and compared favorably with ampicillin and penicillin in achieving bacteriological cure. Cephalothin levels in the central nervous system remained undetectable in both control and infected animals during this time. Tobramycin concentrations were measurable in the CSF, but not in brain tissue in association with an inflammatory stimulus.


Effect of edible chitosan film enriched with anise (Pimpinella anisum L.) essential oil on shelf life and quality of the chicken burger.
In this study, the effect of chitosan film (CF) with different concentrations of anise essential oil AEO (0, 0.5, 1, 1.5 and 2%) on the quality of chicken burger during chilled storage (4 + 1°C) were examined over a period of 12 days. For this purpose, at the first, the physical and mechanical properties of the produced films were studied. Then, the chicken burger was covered with the produced films. Different treatment were analyzed by biochemical properties such as moisture and thiobarbituric acid, bacteriological properties such as total viable counts and total psychrotrophic counts, Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli. The results of this study showed that adding AEO improved the properties of CF, the moisture, solubility, and water vapor permeability decreased in these films. By increasing the concentration of AEO the tensile strength and elasticity of film were increased. CF with AEO, delayed lipid oxidation in chicken burger and improved the chemical properties of chicken burger. Also, microbial spoilage in these samples decreased significantly (p < .05) compared to the control sample. AEO at 2% in all experiments had better results than other treatments (p < .05), and the AEO (1.5% and 2%) had acceptable biochemical, bacteriological attributes up to end of storage, and these treatments could reduce the population of pathogen bacteria below the acceptable level from day 3 until the end of the storage period. Sensory score of the treatment containing AEO at 1.5% was higher than the sensory score of AEO at 2%. Overall, the results of this study showed that the use of CF with AEO as a natural preservative increased the shelf life of meat products. Considering the relatively similar anti-oxidation and antimicrobial effect of AEO at 1.5 and 2% and also economic aspects, optimum dose for AEO could be 1.5% in the film.


Clinical significance of CD155 expression and correlation with cellular components of tumor microenvironment in gastric adenocarcinoma.
CD155 is recently emerging as a promising target in malignancies. However, the relationship between CD155 expression and tumor microenvironment (TME) cell infiltration in gastric adenocarcinoma (GAC) has rarely been clarified. We measured CD155 expression in specimens of gastric precancerous disease and GAC by immunohistochemistry. The association of CD155 expression with GAC progression and cells infiltration in TME was evaluated through 268 GAC tissues and public dataset analysis. We showed that the expression of CD155 was positively correlated with the pathological development of gastric precancerous disease (r = 0.521, P < 0.0001). GAC patients with high CD155 expression had a poorer overall survival (P = 0.033). Moreover, CD155 expression correlated with aggressive clinicopathological features including tumor volume, tumor stage, lymph node involvement, and cell proliferation (P <0.05). Remarkably, CD155 expression positively related to the infiltration of CD68+ macrophages in TME (P = 0.011). Meanwhile, the positive correlation was observed between CD155 and CD31 (P = 0.026). In addition, patients with high CD155 expression combined with low CD3, CD4, CD8, IL-17, IFN-γ or CD19 expression as well as those with high CD155 and α-SMA expression showed significantly worse overall survival (P < 0.05). CD155 may play a pivotal role in the development of GAC through both immunological and non-immunological mechanisms and be expected to become a novel target of immunotherapy in GAC patients.


[Primary gastrointestinal lymphoma].
The gastrointestinal tract is the most common extranodal site for non-Hodgkin's lymphoma. Primary gastrointestinal lymphoma (PGIL) represents a specific entity and has to be differentiated from nodal lymphomas because of its different biological behaviour, prognosis and treatment. Imaging methods are important for detection and staging of PGIL. Gastrointestinal lymphomas produce a spectrum of pathological and radiological appearances. PGIL is encountered often enough to warrant familiarity with the clinical, radiological and therapeutic options.


Family-centered care and the anticipated death of a newborn.
Providing support for a family whose infant is expected to die requires planning and coordinated intervention by members of the health care team. From diagnosis until the death of the infant, the family's choices and needs must be integrated into an acceptable plan of care. The role of the health care team in this process includes four principal phases: planning for the baby's birth, preparing the siblings, caring for the baby, and supporting the family both at the time of the death and afterward. Specific strategies used by physicians, nurses, and child life specialists can encourage the family's full participation in the birth and death of the infant. A description of one family's experiences highlights implications for family-centered practice.


Further thoughts on mast cells, calcium channels and histamine release.
Calcium movements across membranes of mast cells represent a major target for effective anti-allergic drugs, as these are essential events linking stimulation to secretion. Using isolated peritoneal mast cells from two types of genetically different rats and a blocker of the entry of calcium ions into cells, histamine release induced by antigen, calcium ionophore and potassium (in a lowered calcium medium) has been followed. Lower concentrations of the calcium entry blocker were required to inhibit potassium-induced release than those needed for release induced by allergic and other non-allergic stimuli, suggesting that different specialised calcium channels are involved in histamine release from rat mast cells.


Interpreting challenge data from early phase malaria blood stage vaccine trials.
As the quest for an effective blood stage malaria vaccine continues, there is increasing reliance on the use of controlled human malaria infections (CHMI) in non-endemic settings to test vaccine efficacy at the earliest possible time. This is seen as a way to accelerate vaccine research and quickly eliminate candidates with poor efficacy. Areas covered: The data from these studies need to be carefully examined and interpreted in light of the very different roles that antibody and cellular immunity play in protection and within the context of the distinct clinical sensitivities of volunteers living in malaria-non-endemic countries compared to those living in endemic countries. With current strategies, it is likely that vaccines with protective immunological 'signatures' will be missed and potentially good candidates discarded. Expert commentary: Efficacy data from early phase vaccine trials in non-endemic countries should not be used to decide whether or not to proceed to vaccine trials in endemic countries.


The assessment of lower face morphology changes in edentulous patients after prosthodontic rehabilitation, using two methods of measurement.
Using two measurement methods (a three-dimensional laser scanning system and a digital caliper), this study compares the lower face morphology of complete edentulous patients, before and after prosthodontic rehabilitation with bimaxillary complete dentures. Fourteen edentulous patients were randomly selected from the Department of Prosthodontics, at the Faculty of Dental Medicine, "Victor Babes" University of Medicine and Pharmacy, Timisoara, Romania. The changes that occurred in the lower third of the face after prosthodontic treatment were assessed quantitatively by measuring the vertical projection of the distances between two sets of anthropometric landmarks: Subnasale - cutaneous Pogonion (D1) and Labiale superius - Labiale inferius (D2). A two-way repeated measures ANOVA model design was carried out to test for significant interactions, main effects and differences between the two types of measuring devices and between the initial and final rehabilitation time points. The main effect of the type of measuring device showed no statistically significant differences in the measured distances (p=0.24 for D1 and p=0.39 for D2), between the initial and the final rehabilitation time points. Regarding the main effect of time, there were statistically significant differences in both the measured distances D1 and D2 (p=0.001), between the initial and the final rehabilitation time points. The two methods of measurement were equally reliable in the assessment of lower face morphology changes in edentulous patients after prosthodontic rehabilitation with bimaxillary complete dentures. The differences between the measurements taken before and after prosthodontic rehabilitation proved to be statistically significant.


Nanofabrication of heteromolecular organic nanostructures on epitaxial graphene via room temperature feedback-controlled lithography.
Nanoscale control of surface chemistry holds promise for tailoring the electronic, optical, and chemical properties of graphene. Toward this end, the nanofabrication of sub-5-nm heteromolecular organic nanostructures is demonstrated on epitaxial graphene using room temperature ultrahigh vacuum scanning tunneling microscopy. In particular, monolayers of the organic semiconductor 3,4,9,10-perylene-tetracarboxylic dianhydride (PTCDA) are nanopatterned on epitaxial graphene using feedback-controlled lithography (FCL) and then used as chemical resists to template the deposition of N,N'-dioctyl-3,4,9,10-perylene-tetracarboxylic diimide (PTCDI-C8). The generality of this FCL-based nanofabrication procedure suggests its applicability to a wide range of fundamental studies and prototype device fabrication on chemically functionalized graphene.


Appropriate and Constant Potassium Supply Promotes the Growth of M9T337 Apple Rootstocks by Regulating Endogenous Hormones and Carbon and Nitrogen Metabolism.
Potassium (K) is an indispensable nutrient element in the development of fruit trees in terms of yield and quality. It is unclear how a stable or unstable supply of K affects plant growth. We studied the root morphology and physiological and molecular changes in the carbon and nitrogen metabolism of M9T337 apple rootstock under different K levels and supply methods using hydroponics. Five K supply treatments were implemented: continuous low K (KL), initial low and then high K (KLH), appropriate and constant K (KAC), initial high and then low K (KHL), and continuous high K (KH). The results showed that the biomass, root activity, photosynthesis, and carbon and nitrogen metabolism of the M9T337 rootstocks were inhibited under KL, KH, KLH and KHL conditions. The KAC treatment promoted root growth by optimizing endogenous hormone content, enhancing carbon and nitrogen metabolism enzyme activities, improving photosynthesis, optimizing the distribution of carbon and nitrogen, and upregulating the transcription levels of nitrogen assimilation-related genes (nitrate reductase, glutamine synthetase, glutamate synthase, MdNRT1.1, MdNRT1.2, MdNRT1.5, MdNRT2.4). These results suggest that an appropriate and constant K supply ensures the efficient assimilation and utilization of nitrogen and carbon.


Renner-Teller interactions in the dissociative recombination of HCO+.
The formalism developed in the preceding paper for vibrational autoionization via Renner-Teller active vibrations is adapted to treat dissociative recombination and applied to the reaction of HCO(+)+e(-). Existing spectroscopic data on the rovibrational structure of the HCO(+) (2)Sigma(+) ion and the HCO 3ppi (2)Pi Rydberg state are fitted by using the semirigid bender model to extract the parameters required to calculate the autoionization and electron capture widths. The results of this simple model are in good agreement with more detailed first principles calculations of the dissociative recombination cross section and confirm the earlier conclusion that coupling due to the Renner-Teller interaction is largely responsible for the observed dissociative recombination cross section at electron energies below approximately 0.1 eV.


Lack of Efficacy of Pegylated Interferon Monotherapy for Hepatitis C in Patients With End-Stage Renal Disease on Dialysis.
Objectives To evaluate the efficacy, safety and tolerability of pegylated interferon monotherapy for chronic hepatitis C virus (HCV) in patients who are on dialysis. Methods From the University of Chicago Clinical Hepatology Database dated May 2001 to July 2005, 13 patients on dialysis with hepatitis C who have been treated with pegylated interferon were identified. Demographic and laboratory data were obtained from medical records. Patients received pegylated interferon alfa-2a at 135 µg subcutaneous (SQ) weekly (n = 8) or pegylated interferon alfa-2b at 1 µg/kg SQ weekly (n = 5). Side effects from the medication were noted. Results There were 7 men and 6 women, with a mean age of 54±11 years; 11 patients (85%) were African American and 11 patients (85%) were infected with HCV genotype 1. The median serum HCV RNA level was 3,273,000 copies/mL (range, 207,000 to >40,000,000), and the median serum alanine aminotransferase level was 29 IU/mL (range, 19-77). Four patients (30%) had bridging fibrosis or cirrhosis on liver biopsy. None of the 13 patients achieved sustained virologic response; 2 patients (15%) had an undetectable viral load at the end of therapy but relapsed within 6 months of follow-up. The most common side effects were fatigue (100%), anemia defined as 2 g/dL or greater drop in hemoglobin level (60%), and psychiatric symptoms (30%). Conclusions Pegylated interferon is ineffective for HCV infection in patients on dialysis. Furthermore, worsening anemia, which is usually prevalent at baseline in dialysis patients, is a common adverse event even in the absence of ribavirin use.


Clefts in quintuplets--a case report.
A report of clefting in three siblings of quintuplets, following treatment with Pergonal, is presented. It is suggested that the varying degrees of the abnormality may be related to cramped intra-uterine conditons or differing pressures at a critical developmental stage. To our knowledge, no previous reports of congential abnormalities associated with Pergonal therapy exist, and we feel that the causal relationship should be explored using a suitable animal model.


A weekly alternating diet between caloric restriction and medium fat protects the liver from fatty liver development in middle-aged C57BL/6J mice.
We investigated whether a novel dietary intervention consisting of an every-other-week calorie-restricted diet could prevent nonalcoholic fatty liver disease (NAFLD) development induced by a medium-fat (MF) diet. Nine-week-old male C57BL/6J mice received either a (i) control (C), (ii) 30E% calorie restricted (CR), (iii) MF (25E% fat), or (iv) intermittent (INT) diet, a diet alternating weekly between 40E% CR and an ad libitum MF diet until sacrifice at the age of 12 months. The metabolic, morphological, and molecular features of NAFLD were examined. The INT diet resulted in healthy metabolic and morphological features as displayed by the continuous CR diet: glucose tolerant, low hepatic triglyceride content, low plasma alanine aminotransferase. In contrast, the C- and MF-exposed mice with high body weight developed signs of NAFLD. However, the gene expression profiles of INT-exposed mice differed to those of CR-exposed mice and showed to be more similar with those of C- and MF-exposed mice with a comparable body weight. Our study reveals that the INT diet maintains metabolic health and reverses the adverse effects of the MF diet, thus effectively prevents the development of NAFLD in 12-month-old male C57BL/6J mice.


Putting the Community back into Community Health Needs Assessments: Maximizing Partnerships Via Community-Based Participatory Research.
The community health needs assessment (CHNA) mandate of the Patient Protection and Affordable Care Act (ACA) has the potential to make significant and sustainable change in the health of communities. However, to date many hospital-led assessments have used traditional, top-down data collection approaches that overemphasize individualized community member deficits and underutilize collaboration across sectors. The purpose of this paper is to present the principles of community-based participatory research (CBPR) as a framework for conducting CHNAs in a way that mitigates the potential for harm, waste, and misrepresentation of community assets and needs that characterizes many existing CHNA processes, illustrating the power of applying CBPR partnerships to this process. CBPR is a framework to engage community members directly in research design, the collection and analysis of data, and the creation of action plans that address research findings. Key principles include collaborative involvement, establishment of empowering processes, and long-term commitment. A case example of an innovative community partnership demonstrates the power and challenges of taking a CBPR approach to the CHNA process. CBPR has incredible potential to be incorporated into ACA-mandated hospital CHNAs, leading to increased impact and shared power with community members.


Central Pontine Myelinolysis and Hypokalemic Paralysis as Presenting Manifestations of Sjogren's Syndrome.
Neurological involvement in Sjogren's syndrome can have varied manifestations and can precede the classical sicca symptoms of Sjogren's syndrome. A 32-year-old woman presented with acute quadriparesis and dysarthria. She had severe hypokalemia, and an MRI of the brain showed a lesion in the central pons that was hyperintense on T2 and fluid-attenuated inversion recovery (FLAIR) sequences sparing the periphery, a trident appearance characteristic of central pontine myelinolysis (CPM). On further evaluation, she was found to have distal renal tubular acidosis (dRTA) due to primary Sjogren's syndrome. She was treated with steroids and other supportive measures, and she recovered completely in one month. We describe a mild form of CPM with classical MRI features in a patient with Sjogren's syndrome and hypokalemia due to dRTA.


Lead poisoning: hair analysis shows the calendar of events.
A woman was given frequent oral doses of lead nitrate over an extended time period. Diagnosis was finally made about 6 months after the first symptoms appeared. Analysis of hair taken at 2 different times was used as evidence in court. The hair lead concentrations exhibited rapid changes and normalised shortly after chelation treatment had been instituted. This finding suggests that the hair lead level may reflect a current absorption level rather than the lead content of whole blood which, in this case, changed much more slowly. Despite the extremely high blood lead levels and the severe clinical lead poisoning, lead concentrations in hair were relatively low in comparison to the concentrations found in hair of workers exposed to high levels of airborne lead. This observation suggests that a large proportion of the lead content of hair from individuals with respiratory lead exposure may be exogenous.


Apoptosis in activated rat pancreatic stellate cells.
Proliferation and matrix synthesis by activated pancreatic stellate cells (PSC) participate in the development of chronic pancreatitis. Apoptosis of PSC may terminate this process but has not yet been studied in this particular cell type and was the aim of the present study. PSC were isolated from rat pancreas and characterized for expression of glial fibrillary acidic protein, alpha-smooth muscle actin, CD95, and tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) receptors. Apoptosis was determined by TdT-UTP nick end-labeling reaction, annexin V binding, and caspase-8 activation. Both CD95L and TRAIL induced apoptosis in PSC. The apoptotic response was minor in PSC cultured for 7 days but increased markedly thereafter. Sensitization of PSC with culture duration was accompanied by increased expression of CD95 and TRAIL receptor 2 and no alterations of Flip expression or protein kinase B phosphorylation but was paralleled by the appearance of a COOH-terminal cleavage product of receptor-interacting protein. PSC apoptosis was also induced by PK-11195, a ligand of the peripheral benzodiazepine receptor. PSC apoptosis may be important in terminating the wound-healing response after pancreas injury and exhibits features distinct from apoptosis induction in hepatic stellate cells.


Study protocol for a prospective cohort study examining the predictive potential of dynamic symptom networks for the onset and progression of psychosis: the Mapping Individual Routes of Risk and Resilience (Mirorr) study.
Our current ability to predict the course and outcome of early psychotic symptoms is limited, hampering timely treatment. To improve our understanding of the development of psychosis, a different approach to psychopathology may be productive. We propose to reconceptualise psychopathology from a network perspective, according to which symptoms act as a dynamic, interconnected system, impacting on each other over time and across diagnostic boundaries to form symptom networks. Adopting this network approach, the Mapping Individual Routes of Risk and Resilience study aims to determine whether characteristics of symptom networks can predict illness course and outcome of early psychotic symptoms. The sample consists of n=100 participants aged 18-35 years, divided into four subgroups (n=4×25) with increasing levels of severity of psychopathology, representing successive stages of clinical progression. Individuals representing the initial stage have a relatively low expression of psychotic experiences (general population), whereas individuals representing the end stage are help seeking and display a psychometric expression of psychosis, putting them at ultra-high risk for transition to psychotic disorder. At baseline and 1-year follow-up, participants report their symptoms, affective states and experiences for three consecutive months in short, daily questionnaires on their smartphone, which will be used to map individual networks. Network parameters, including the strength and directionality of symptom connections and centrality indices, will be estimated and associated to individual differences in and within-individual progression through stages of clinical severity and functioning over the next 3 years. The study has been approved by the local medical ethical committee (ABR no. NL52974.042.15). The results of the study will be published in (inter)national peer-reviewed journals, presented at research, clinical and general public conferences. The results will assist in improving and fine-tuning dynamic models of psychopathology, stimulating both clinical and scientific progress. NTR6205 ; Pre-results.


Accelerated active phase transformation of NiO powered by Pt single atoms for enhanced oxygen evolution reaction.
Phase transformation of electrode materials widely occurs in electrocatalytic reactions. Metal oxides are promising electrocatalysts for the oxygen evolution reaction (OER); their phase transformation is a key step for the multi-electron OER, and requires extra overpotential. However, little attention has been paid to accelerating and enhancing the phase transformation. Here, we report for the first time that single-atom Pt incorporated into the bulk crystalline phase of porous NiO nanocubes (0.5 wt% Pt/NiO) can greatly promote the active phase (NiOOH) evolution. The Pt doping was achieved by a scalable nanocasting approach using SiO2 as the hard template. In comparison with Pt/NiO samples with PtO2 nanoparticles segregated at the NiO surface (1 wt% Pt), as well as atomistic Pt atoms solely bound at the surface by atomic layer deposition, the bulk Pt doping shows the strongest power in facilitating active phase transformation, which leads to improved OER activity with reduced overpotential and Tafel slope. Experiential data revealed that the charge-transfer from Pt to Ni through O leads to a local weaker Ni-O bond. First principles calculations confirmed that rather than acting as an active site for the OER, monatomic Pt effectively increases the phase transformation rate by reducing the migration barrier of nearby Ni atoms. Our discoveries reveal the relationships of the heteroatom doped structure and phase transformation behavior during the electrochemical process and offer a new route for designing high-performance electrocatalysts.


Inhibition of root elongation by ethylene in wetland and non-wetland plant species and the impact of longitudinal ventilation.
The slow gas diffusion rate in flooded soil not only causes oxygen deficiency, but also favours the accumulation of ethylene in root systems to concentrations that may strongly affect root elongation. Previously published experiments showed that root elongation in rice is much less strongly inhibited by ethylene than in some other species less well adapted to wet conditions. Rice roots have also been reported to produce abnormally little ethylene. We tested if these traits are typical of wetland species and are thus likely to be widespread adaptive traits. Comparisons using 14 species indicated that insensitivity to the inhibiting effects of ethylene on root elongation is unlikely to be a common feature of temperate wetland species. However, resistance to longitudinal gas diffusion within roots of wetland species, which largely depends on diameter and the presence of gas-filled channels, was found to be less than in non-wetland species. We show that this can help maintain low internal ethylene concentrations by venting accumulated gas to the shoot and atmosphere.


[Treatment of chronic obstructive respiratory disease with a new purified theophylline preparation in delayed-action form].
The treatment of chronic obstructive respiratory diseases with sustained release theophylline is steadily gaining in importance. Body plethysmography is applied for evaluating the extent of bronchospasmolysis during therapy of a new sustained release formula. A traditional prolonged-action theophylline-ethylenediamine preparation was administered to obtain comparative data. The sustained release formula containing pure theophylline meets the standards of a modern theophylline therapy. The graduation of the film tablet allows individualized dosage without losing the sustained release action by slowly increasing the theophylline-serum-level during a dosage interval of 12 hours.


A keratin 10 gene mutation (Arg156Cys) in a Japanese patient with bullous congenital ichthyosiform erythroderma.
We described a 19-year old Japanese female with bullous congenital ichthyosiform erythroderma (BCIE) and examined the keratin gene mutation. Physical examination disclosed generalized erythema, ichthyosiform skin with scales, and erosions without palmoplantar keratoderma. Histological examination revealed hyperkeratosis with vacuolar degeneration in the granular layer of the epidermis. Sequence analysis demonstrated a C to G transition at the first position of codon 156 in the keratin 10 gene. The amino acid at codon 156 was deduced to have changed from arginine to cystine. Substitution from arginine to cysteine at codon 156 of the K 10 gene is assumed to be fatal for keratin filament assembly regardless of racial or ethnic difference.


Target-mask shape congruence impacts the type of metacontrast masking.
Visual metacontrast masking may depend on the time intervals between target and mask in two qualitatively different ways: in type-A masking the smaller the mask delay from target the stronger the masking while in type-B masking maximal masking effect is obtained with a larger temporal delay of the mask. Variability in the qualitative apperance of masking functions has been explained by variability in stimuli parameters and tasks. Recent research on metacontrast masking has surprisingly shown that both of these types of functions can be found with an identical range of stimulation parameters depending on individual differences between observers. Here we show that obtaining clear-cut type-A masking depends on whether target and mask shapes are congruent or incongruent and whether observers use the cues available due to the congruence factor. Conspicuously expressed type-A masking is selectively associated with incongruent target-mask pairings. In the latter conditions target identification level significantly drops with the shortest target-to-mask delays.


Role of Computationally Evaluated Target Specificity in the Hepatotoxicity of Gapmer Antisense Oligonucleotides.
Gapmer antisense oligonucleotides (gapmers) sometimes cleave nontarget pre-mRNAs by recognizing target-like intronic/exonic portions. This off-target RNA cleavage could be a major cause of the hepatotoxicity that is induced by gapmers. In line with these findings, we hypothesized that gapmers with higher specificity have less hepatotoxicity, and that those with lower specificity have greater toxicity. To examine this concept, we investigated various Malat1-targeting gapmers with various computationally evaluated target specificities. We had expected that higher specificity gapmers would have lower hepatotoxicity, but these factors were not significantly related. In silico analysis of gapmer sequences does not always contribute to mitigating the risk of hepatotoxicity. Transcriptome analysis indicated that nontoxic gapmers do not cleave off-target RNAs, although they have many target-like RNA sequences. The present results shed light on the mechanism of the hepatotoxicity of gapmers.


2-deoxy-D-glucose enhances the cytotoxicity of topoisomerase inhibitors in human tumor cell lines.
Resistance of tumors due to restricted drug accumulation and reversal of DNA lesions in tumor cells as well as normal tissue toxicity limit the efficacy of topoisomerase inhibition based anticancer drugs. It has been proposed that selective inhibition of energy dependent repair processes and enhanced retention of drug in cancer cells can significantly improve the therapeutic efficacy. The purpose of these studies was to verify this proposition by investigating the effects of 2-deoxy-D-glucose (2-DG) an inhibitor of the glycolytic ATP production on the cytotoxicity of certain topoisomerase inhibitors in human tumor cell lines. Human glioma (BMG-1 and U-87) and squamous carcinoma (4197 and 4451) cell lines were investigated with topo-poisons like etoposide (topo II inhibitor), camptothecin (topo I inhibitor) and hoechst-33342 (topo I and II inhibitor). DNA damage induction (halo assay), cell survival (macro colony assay), cytogenetic damage (micronuclei) and apoptosis (morphological analysis) were investigated. Presence of 2-DG for 2 h following exposure to the topoisomerase inhibitors enhanced the cell death (macro colony assay) in a concentration dependent manner and a 2-3-fold increase was observed at 5 mM (equimolar with glucose). Halo assay revealed that 2-DG inhibited the reversal of cleavable complex leading to the accumulation of DNA strand breaks. Under these conditions 2-DG enhanced the drug-induced micronuclei formation by nearly 2 folds with etoposide and hoechst-33342 and a 4-fold increase in delayed apoptosis was observed in case of etoposide. These results clearly demonstrate that presence of 2-DG for a few hours following exposure to topo-inhibitors enhances the cytotoxicity, primarily by increasing the cytogenetic damage.


The consequences of attention-deficit/hyperactivity disorder in adults.
Until recently, attention-deficit/hyperactivity disorder (ADHD) was a diagnosis reserved for children and adolescents as it was believed to dissipate before adulthood. New evidence, however, supports the persistence of ADHD beyond adolescence, and it is now recognized as a chronic neurobehavioral disorder in adults. Adults with ADHD have difficulties with school, work, family interactions, and social activities. Although treatments are available for adult ADHD, many patients never receive an accurate diagnosis that would afford them appropriate therapeutic intervention. If left untreated, adult ADHD can cause significant personal, social, and economic burdens that can have a negative impact on overall quality of life. This article discusses how ADHD presents in adults and the effects of the disorder on educational, occupational, interpersonal, and social functioning. Currently available treatments for ADHD in adults are also reviewed.


Retinoic acid induced heparin-binding protein expression and localization during gastrulation, neurulation, and organogenesis.
Retinoic acid induced heparin-binding protein (RIHB) is a highly basic, soluble polypeptide of the chick embryonic extracellular matrix. We have examined the expression and localization of RIHB during very early embryogenesis by in situ hybridization and immunohistochemistry. RIHB mRNA is very weakly detectable above background in the blastodiscs of unincubated eggs. The expression increases greatly over the first 24 hours of incubation, and is observed throughout the blastodisc in all three of the germ layers following gastrulation. As neurulation occurs, the expression becomes more restricted to certain areas, notably the ectoderm, the neural folds, and especially the notochord. After the neural tube has formed the expression in the tube itself decreases dramatically, whereas the expression in the head ectoderm and the notochord persists. After 72 hours of incubation expression remains relatively high throughout most of the embryo, with higher levels of expression in regions undergoing organogenesis and lower levels in organs which have already differentiated. RIHB protein is also weakly detectable in unincubated eggs as patches of immunoreactive material between the blastodisc and the vitelline. After 6 hours of incubation small regions of basement membrane are immunoreactive. RIHB is detected in this matrix, apparently before even fibronectin. The amount of RIHB protein increases dramatically over the first 24 hours of incubation. It is found in basement membrane separating the epiblast from the hypoblast, then later in that separating the ectoderm from the mesoderm. It is also detected surrounding individual cells, especially of the ectodermal layer. During neurulation RIHB is observed in the basement membrane surrounding the neural fold and the notochord, and in the lamina separating the ectodermal, mesodermal, and endodermal layers. Later in development, RIHB is detected in the basement membrane under the epidermis, throughout the developing limbs, and in the lamina of various developing organs, such as the eye, the pulmonary bud, the intestine, and the mesonephros. These results demonstrate that RIHB is highly expressed during the early embryonic period, by all three germ layers, and is an important and very early component of the embryonic extracellular matrix. Its very broad expression and localization argue for a more general role in development than its demonstrated weak neurotrophic activity.


Fine structure of cell division in Chlamydomonas reinhardi. Basal bodies and microtubules.
Cell division in log-phase cultures of the unicellular, biflagellate alga, Chlamydomonas reinhardi, has been studied with the electron microscope. The two basal bodies of the cell replicate prior to cytokinesis; stages in basal body formation are presented. At the time of cell division, the original basal bodies detach from the flagella, and the four basal bodies appear to be involved in the orientation of the plane of the cleavage furrow. Four sets of microtubules participate in cell division. Spindle microtubules are involved in a mitosis that is marked by the presence of an intact nuclear envelope. A band of microtubules arcs over the mitotic nucleus, indicating the future cleavage plane. A third set of microtubules appears between the daughter nuclei at telophase, and microtubules comprising the "cleavage apparatus" radiate from the basal bodies and extend along both sides of the cleavage furrow during cytokinesis. Features of cell division in C. reinhardi are discussed and related to cell division in other organisms. It is proposed that microtubules participate in the formation of the cleavage furrow in C. reinhardi.


Immunohistochemical localization of insulin-like growth factor 1 receptors in benign and malignant tissues of the female genital tract.
The distribution of insulin-like growth factor I (IGF-1) receptors in the female genital tract was examined by immunohistochemistry. The monoclonal antibody alpha-IR-3, which binds to the alpha-subunits of the IGF-1 receptor, was used for specific binding and the peroxidase-antiperoxidase method was used for staining. IGF-1 receptors were consistently detected in the epithelium of cervix, endometrium and the fallopian tube. Furthermore, high expression of the IGF-1 receptors was found in ovarian cancer tissue, where predominantly the stromal cells around the vessels gave an intense staining. Since the expression of the IGF-1 receptors in tumor epithelium was only weak and inconsistent, it is tempting to speculate that the stromal compartment in ovarian cancer is the target tissue for the effects of IGF-1.