Morphologic and molecular study of hemoparasites in wild corvids and evidence of sequence identity with Plasmodium DNA detected in captive black-footed penguins (Spheniscus demersus).
A morphologic and molecular epidemiologic investigation was conducted on a captive African black-footed penguin (Spheniscus demersus) colony with a history of Plasmodium infections at La Palmyre Zoo (France). Each penguin received 12.5 mg of pyrimethamine twice a week as a prophylaxis every year from April to November. Although Plasmodium parasites were not detected in blood smears and tissues collected from the penguins, various blood parasites were recorded in blood smears from wild Eurasian magpies (Pica pica) and carrion crows (Corvus corone) sampled at the same time in the study area. These parasites consisted of several Plasmodium spp. (P. lenoblei, P. dorsti, P bioccai, P. relictum, P. dherteae, P. beaucournui, P. maior, P. tranieri, and P. snounoui), Parahaemoproteus spp., Trypanosoma spp., and Leucocytozoon spp. On the other hand, nested polymerase chain reaction enabled detection of Plasmodium DNA in 28/44 (64%) penguins, 15/25 (60%) magpies, and 4/9 (44%) crows. Sequencing and phylogenetic analyses indicated that the parasite DNA amplified from the penguins, magpies, and crows were similar. Magpies and crows could therefore act as a reservoir for penguin Plasmodium infections, which may be more prevalent than previously thought. Morphologic characterization of the Plasmodium spp. detected in the penguins, as well as further biological and epidemiologic studies, are needed to fully understand the transmission of Plasmodium parasites to captive penguins.


Effective enrichment and mass spectrometry analysis of phosphopeptides using mesoporous metal oxide nanomaterials.
Mass spectrometry (MS)-based phosphoproteomics remains challenging due to the low abundance of phosphoproteins and substoichiometric phosphorylation. This demands better methods to effectively enrich phosphoproteins/peptides prior to MS analysis. We have previously communicated the first use of mesoporous zirconium dioxide (ZrO(2)) nanomaterials for effective phosphopeptide enrichment. Here, we present the full report including the synthesis, characterization, and application of mesoporous titanium dioxide (TiO(2)), ZrO(2), and hafnium dioxide (HfO(2)) in phosphopeptide enrichment and MS analysis. Mesoporous ZrO(2) and HfO(2) are demonstrated to be superior to TiO(2) for phosphopeptide enrichment from a complex mixture with high specificity (>99%), which could almost be considered as a "purification", mainly because of the extremely large active surface area of mesoporous nanomaterials. A single enrichment and Fourier transform MS analysis of phosphopeptides digested from a complex mixture containing 7% of alpha-casein identified 21 out of 22 phosphorylation sites for alpha-casein. Moreover, the mesoporous ZrO(2) and HfO(2) can be reused after a simple solution regeneration procedure with comparable enrichment performance to that of fresh materials. Mesoporous ZrO(2) and HfO(2) nanomaterials hold great promise for applications in MS-based phosphoproteomics.


There is no correlation between glucocorticoid receptor mRNA expression and protein binding in the brains of house sparrows (Passer domesticus).
The stress response represents an animal's attempt to cope with a noxious stimulus through a rapid release of corticosterone or cortisol (CORT) into the bloodstream, resulting in a suite of physiological and behavioral changes. These changes are mediated in large part through CORT's binding to two different intracellular receptors, the high-affinity mineralocorticoid receptor (MR) and the lower-affinity glucocorticoid receptor (GR). We tested the hypothesis that GR and MR mRNA expression would correlate with functional protein expression in neuronal tissue of wild-caught house sparrows (Passer domesticus). To test this hypothesis, we performed a parallel procedure in which protein concentrations were quantified in one half of house sparrow brains (n=16) using radioligand binding assays, and mRNA levels were quantified in the other brain half using reverse-transcriptase quantitative PCR (RT-qPCR). Two reference genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and TATA-box binding protein (TBP), were used for relative quantification of GR and MR mRNA. Quantifications showed that these two reference genes were not correlated with each other. Furthermore, there was no correlation between mRNA and protein levels for GR or MR using either reference gene, suggesting that regulation of mRNA and protein levels for MR and GR is not tightly linked. This study provides insight into the importance of regulatory steps between mRNA expression and the creation and stability of a functional protein. The overall conclusion is that mRNA expression cannot be used as a proxy for GR or MR binding in house sparrows.


Photocontrol of the Functional Coupling between Photosynthesis and Stomatal Conductance in the Intact Leaf : Blue Light and Par-Dependent Photosystems in Guard Cells.
The photocontrol of the functional coupling between photosynthesis and stomatal conductance in the leaf was investigated in gas exchange experiments using monochromatic light provided by lasers. Net photosynthesis and stomatal conductance were measured in attached leaves of Malva parviflora L. as a function of photon irradiance at 457.9 and 640.0 nanometers.Photosynthetic rates and quantum yields of photosynthesis were higher under red light than under blue, on an absorbed or incident basis.Stomatal conductance was higher under blue than under red light at all intensities. Based on a calculated apparent photon efficiency of conductance, blue and red light had similar effects on conductance at intensities higher than 0.02 millimoles per square meter per second, but blue light was several-fold more efficient at very low photon irradiances. Red light had no effect on conductance at photon irradiances below 0.02 millimoles per square meter per second. These observations support the hypothesis that stomatal conductance is modulated by two photosystems: a blue light-dependent one, driving stomatal opening at low light intensities and a photosynthetically active radiation (PAR)-dependent one operating at higher irradiances.When low intensity blue light was used to illuminate a leaf already irradiated with high intensity, 640 nanometers light, the leaf exhibited substantial increases in stomatal conductance. Net photosynthesis changed only slightly. Additional far-red light increased net photosynthesis without affecting stomatal conductance. These observations indicate that under conditions where the PAR-dependent system is driven by high intensity red light, the blue light-dependent system has an additive effect on stomatal conductance.The wavelength dependence of photosynthesis and stomatal conductance demonstrates that these processes are not obligatorily coupled and can be controlled by light, independent of prevailing levels of intercellular CO(2). The blue light-dependent system in the guard cells may function as a specific light sensor while the PAR-dependent system supplies a CO(2)-modulated energy source providing functional coupling between the guard cells and the photosynthesizing mesophyll.


[Type I and IV collagenases and their endogenous regulators in immortalized and transformed fibroblasts].
To elucidate the role of matrix metalloproteinases (MMP) in carcinogenesis, the expression of collagenases of types I (MMP-I) and IV (MMP-2 and MMP-9) as well as the behaviour of urokinase-like plasminogen activator (uPA) and of tissue MMP inhibitors (TIMP) in immortalized (IF) and transformed (TF) fibroblasts were investigated. The study was carried out using embryo rat fibroblasts, sequentially immortalized with the LT gene of human papilloma virus and transformed with the E7 gene of human papilloma virus (HPV-16). As control was used the primary fibroblast (PF) culture of Fisher rats. In IF, the collagenase activity was at the same level as it was in PF. The activity of uPA in IF was increased by 2-2.5-fold; the titrated amount of free endogenous inhibitors in IF and PF was at essentially the same level while being markedly higher than in TF. At the stage of fibroblast transformation with the E7 gene of HPV-16, there was seen an increase of Type IV collagenases and a decrease of Type I collagenase, both these indices being most pronounced in the cells with most developed tumorigenic properties. In TF there occurred a decrease of free endogenous MMP inhibitors relative to the enzyme activity and, at the same time, a decrease in uAF activity, indicating the changes occurring in the enzyme/inhibitor/activator ratio and hence the enhancement of the destructive potential of the cells (in this case, at the cost of Type IV collagenase activity).


[Three cases of hydronephrosis in which the indwelt ureteral stent was ineffective. Urodynamic studies using a stent indwelling model for retroperitoneal fibrosis].
We report 3 cases in which ureteral stents were indwelt to treat hydronephrosis due to retroperitoneal fibrosis, resulting in aggravation of hydronephrosis and renal function. An urodynamic study was carried out to determine the factors that aggravated hydronephrosis. The subjects were a 59-year-old male with prostatic cancer and bilateral hydronephrosis due to intrapelvic lymph node metastasis, a 49-year-old female with retroperitoneal lymph node metastasis and bilateral hydronephrosis following surgery for stomach cancer, and a 65-year-old male with hydronephrosis due to idiopathic retroperitoneal fibrosis. Indwelling of ureteral stents in the 3 patients resulted in aggravation of hydronephrosis and renal function. Stent indwelling models of retroperitoneal fibrosis were produced using adult mongrel dogs. A pressure flow study was carried out through the nephrostomy in each experimental model to determine the intrapelvic pressure and urine volume. In all stent indwelling models, the urine volume showed changes similar to those in the controls. In the stent indwelling models, the intrapelvic pressure showed a significant increase with an increase in the volume of water instilled through the nephrostomy in the stent indwelling models with retroperitoneal fibrosis, unlike the controls and models with ureteral stenosis. In conclusion, the clinical cases and model experiment suggested that unlike partial ureteral stenosis, in ureters in which extensibility was disturbed in an extensive area, the indwelt stent caused an increase in the resistance of the liquid flowing in the ureter, aggravating hydronephrosis.


Deletion of the budBAC operon in Klebsiella pneumoniae to understand the physiological role of 2,3-butanediol biosynthesis.
Klebsiella pneumoniae is known to produce 2,3-butanediol (2,3-BDO), a valuable chemical. In K. pneumoniae, the 2,3-BDO operon (budBAC) is involved in the production of 2,3-BDO. To observe the physiological role of the 2,3-BDO operon in a mixed acid fermentation, we constructed a budBAC-deleted strain (SGSB109). The production of extracellular metabolites, CO2 emission, carbon distribution, and NADH/NAD(+) balance of SGSB109 were compared with the parent strain (SGSB100). When comparing the carbon distribution at 15 hr, four significant differences were observed: in 2,3-BDO biosynthesis, lactate and acetate production (lactate and acetate production increased 2.3-fold and 4.1-fold in SGSB109 compared to SGSB100), CO2 emission (higher in SGSB100), and carbon substrate uptake (higher in SGSB100). Previous studies on the inactivation of the 2,3-BDO operon were focused on the increase of 1,3-propanediol production. Few studies have been done observing the role of 2,3-BDO biosynthesis. This study provides a prime insight into the role of 2,3-BDO biosynthesis of K. pneumoniae.


PEF based hurdle strategy to control Pichia fermentans, Listeria innocua and Escherichia coli k12 in orange juice.
The combination of pulsed electric fields (PEF) and bacteriocins in a hurdle approach has been reported to enhance microbial inactivation. This study investigates the preservation of orange juice using PEF in combination with nisin (2.5 ppm), natamycin (10 ppm), benzoic acid (BA; 100 ppm), or lactic acid, (LA; 500 ppm). Pichia fermentans, a spoilage yeast frequently isolated from orange juice, Escherichia coli k12 or Listeria innocua were inoculated into sterile orange juice (OJ) with, and without, added preservatives. The antimicrobial activity over time was evaluated relative to an untreated control. The effect of PEF treatment (40 kV/cm, 100 micros; max temperature 56 degrees C) was assessed on its own, and in combination with each antimicrobial. The acidic environment of OJ inactivated E. coli k12 (1.5log reduction) and L. innocua (0.7log reduction) slightly but had no effect on P. fermentans. PEF caused a significant decrease (P<0.05) in the viability of P. fermentans, L. innocua and E. coli k12 achieving reductions of 4.8, 3.7 and 6.3log respectively. Nisin combined with PEF inactivated L. innocua and E. coli k12 in a synergistic manner resulting in a total reduction to 5.6 and 7.9log respectively. A similar synergy was shown between LA and PEF in the inactivation of L. innocua and P. fermentans (6.1 and 7.8log reduction), but not E. coli k12. The BA-PEF combination caused an additive inactivation of P. fermentans, whereas the natamycin-PEF combination against P. fermentans was not significantly different to the effect caused by PEF alone. This study shows that combining PEF with the chosen preservatives, at levels lower than those in current use, can provide greater than 5log reductions of E. coli k12, L. innocua and P. fermentans in OJ. These PEF-bio-preservative combination hurdles could provide the beverage industry with effective non-thermal alternatives to prevent microbial spoilage, and improve the safety of fruit juice.


Diabetes and bone.
Traditionally, patients with type 1 diabetes were regarded to be at an increased risk of fractures whereas type 2 diabetics were assumed to be protected from fractures since many of them have high bone mineral density. Nevertheless, several clinical studies consistently demonstrated that type 2 diabetes is a paradigm of a disease with an increased risk of fractures in the presence of high bone mass. The pathophysiology of decreased bone strength in diabetes mellitus is multifactorial: insulin deficiency, insulin resistance, osteoblast insufficiency, vitamin D deficiency, formation of advanced glycation end-products in bone, and microvascular complications appear to contribute. Drugs used for the treatment of type 2 diabetes also may influence bone fragility: thiazolidinedione use has been associated with an increased risk of fractures.


Coping With Information Style, Self-Concealment, Internalized Stigma, and Family Burden Among Parents of Children With Psychiatric Disorders.
Parents of children with psychiatric disorders who are hospitalized in a psychiatric unit often experience family burden. Family burden has been found to be affected by many variables related to parents' personal traits and ways of reacting to the disorder. The current study examined the association between information coping styles (monitoring and blunting) and family burden, among parents of children who were hospitalized in a day care unit. The possible role of self-stigma as a mediator between coping style and family burden and the role of self-concealment as a moderator between coping style and self-stigma were examined. A total of 41 parents completed questionnaires assessing their levels of self-stigma, information coping style, self-concealment, and family burden. Self-stigma was found to mediate the positive association between the monitoring coping style and family burden. Moreover, a moderation effect of self-concealment was found, indicating that monitoring parents suffered from higher levels of self-stigma particularly if they had a high tendency toward self-concealment. Taking into account parents' information, coping style, self-stigma, and self-concealment can help professionals tailor family interventions according to parents' diverse needs. A monitoring coping style may not be beneficial especially when combined with concealment, suggesting the need to promote other coping styles.


A retrocopy of a gene can functionally displace the source gene in evolution.
The e(y)2 gene of Drosophila melanogaster encodes the ubiquitous evolutionarily conserved co-activator of RNA polymerase II that is involved in transcription regulation of a high number of genes. The Drosophila e(y)2b gene, paralogue of the e(y)2 has been found. The analysis of structure of the e(y)2, e(y)2b and its orthologues from other species reveals that the e(y)2 gene derived as a result of retroposition of the e(y)2b during Drosophila evolution. The mRNA-derived retrogenes lack introns or regulatory regions; most of them become pseudogenes whereas some acquire tissue-specific functions. Here we describe the different situation: the e(y)2 retrogene performs the general function and is ubiquitously expressed, while the source gene is functional only in a small group of male germ cells. This must have resulted from retroposition into a transcriptionally favorable region of the genome.


Reproductive health clinic raided and closed in Bogota, Colombia.
On December 17, armed criminal investigative forces acting under the direction of Colombia's secretary and deputy secretary of health, brutally raided the main clinic of the Orientame reproductive health care federation in Bogota, Colombia. Allegedly responding to charges that the clinic was providing abortions, investigators entered the clinic without warning, brandishing automatic weapons. Once inside, they threatened and physically assaulted patients and clinic staff, destroyed office equipment, broke down doors, and ordered the clinic to close. Since its founding in 1977, Orientame has become Colombia's second largest non-governmental family planning organization. The federation, consisting of three clinics, provides a full range of obstetric, gynecological, and information services, including family planning counseling, cancer screening, treatment for sexually transmitted diseases, prenatal care, adoption placement and counseling, and treatment of incomplete abortion. Serving approximately 250,000 women in its 17 years of operation, Orientame has a sliding fee scale and often waives costs for women who are unable to pay. Were Orientame to close its doors permanently, women facing complications from unsafe abortions but unable to pay private doctors would be forced to go to public hospitals and risk arrest. Despite the country's prohibition of abortions not necessary to save a woman's life, an estimated 288,400 Colombian women underwent abortions in 1989; 57,680 were hospitalized for complications from unsafe or incomplete procedures.


A Concise Asymmetric Total Synthesis of (+)-Epilupinine.
Asymmetric total synthesis of (+)-epilupinine was achieved in just three steps using only commercially available common reagents. The total synthesis involved alkylations of N-nosylamide, ozone oxidation, and sequential reactions of the removal of the nosyl group, intramolecular dehydrative condensation, intramolecular Mannich reaction catalyzed by l-proline, and a reduction.


Bullying in adolescents: role, type of violence and determinants.
To determine the prevalence of bullying and its determinants among adolescents attending school. An analytical cross-sectional study was conducted with a sample of adolescents, who were selected using a multistage probability sampling, from 20 public educational institutions in a Colombian city, in which a simple and multivariate binomial regression was carried out. A total of 500 adolescents participated from which 50.4% were women and 53.2% in their middle adolescence. Verbal violence prevailed in 66.5%, followed by physical violence in 32.0% and social bullying in 30.6%. 69.5% of the adolescents have been witnesses, 35.8% victims, and 14.2% aggressors. 80.5% of the aggressors were victims of bullying. In the multivariate analysis, an association was found between being a victim and having disabilities (PR:2.4; CI: 1.6-3.7), verbal aggression in the home (PR: 1.7; CI: 1.2-2.3) and drug abuse (PR: 1.7; CI 1.1-2.8). Being an aggressor was also associated with being a victim (PR: 7.2; CI 3.6-14.3) and alcohol abuse (PR: 2.2; CI: 1.3-3.8). The frequency of bullying observed and the associated determinants demonstrate the persistence of this problem in adolescents and the need to develop a culture of appropriate and inclusive coexistence that goes beyond the school setting.


Static and rotational output variation of a tomotherapy unit.
A simple device to check the machine output in rotational mode is described. Integral and instantaneous beam output in absolute dose can be measured and analyzed in a very simple manner. A method to correlate the results with the value obtained during the machine calibration in static beam and the reference value used by the treatment planning system for dose calculation in rotation beam is also described. Using this method and device, the daily output can be performed in rotational conditions which are closer to the treatment conditions. Measurement results using this device show differences in the beam output between static and rotational delivery which have to be considered carefully during the calibration process.


Acinetobacter baumannii 2002-2008: increase of carbapenem-associated multiclass resistance in the United States.
The study consisted of data for 55,330 U.S. Acinetobacter baumannii isolates from The Surveillance Network(®) database for the period 2002-2008. Risk factors were time, age, sex, census region, location (Ward or ICU), and isolate source. Antimicrobial susceptibility data were available for carbapenems, cephalosporins, aminoglycosides, fluoroquinolones, and β-lactam/β-lactamase inhibitor combinations. Multiclass resistance was defined as nonsusceptibility to carbapenems and two or more additional classes. Odds of resistance were obtained using a logistic regression model with cubic splines. Carbapenem-associated multiclass resistance has had a 3.7-fold (95% confidence interval [CI] 3.4-4.3) increase from 20.6% in 2002 to 49.2% in 2008. Among blood isolates the increase was by 2.2 times (95% CI 1.7-2.9). Subjects <18 years old had significantly (p < 0.001) lower rates in 2002 (6.9%) than those 65 years or older (21.5%), but by 2008 this difference diminished as rates increased to 44.2% and 54.2%, respectively. A similar divergence was also observed between ICU and Ward, with no differences in 2002, whereas in 2008 ICU isolates had significantly higher rates (55.2%, 95% CI 53.6%-56.9%) than Ward isolates (45.6%, 95% CI 44.2%-47.0%). Over half of all A. baumannii-resistant isolates were carbapenem and multiclass resistant in 2008. Rates among subjects <18 years old have increased faster than those of the elderly, and in the ICU as compared to Ward.


Augmented Reality and Plastic Surgery Training: A Qualitative Study.
Background and objective The coronavirus disease 2019 (COVID-19) pandemic has led to many challenges in face-to-face teaching and training in plastic surgery. However, it has also proved to be an incubator for many technological solutions. Augmented reality (AR) platforms may offer a safe, equitable, and efficient means to provide training in plastic surgery. This study aimed to explore the user's experience of AR as an educational intervention during the COVID-19 pandemic in the United Kingdom (UK). Materials and methods The Proximie® AR platform (Proximie Limited, London, UK) has been in use in a UK plastic surgical department for facilitating webinars, visual libraries, and streamed procedures. The experience of a range of trainers and trainees was qualitatively explored through 10 individual interviews. Data-emergent theme analysis was also performed. Results AR was well-received in the context of COVID-19 and training in general as a means to enable theatre access, and visual revision, remotely. The potential for its use in remote coaching and telementoring was also discussed. Recommendations were made by the users to optimise the experience both from the trainer and learner perspectives. Data were presented pertaining to the following themes: surgical AR as a substitute for hands-on learning; surgical AR and theoretical learning; considerations specific to streamed procedures using Proximie®; considerations in the use of technology in general. Conclusion Harnessing novel technologies in surgical education offers an exciting opportunity, fast-tracked by COVID-19, but applicable beyond it. Though this study includes a small sample size, its findings suggest that AR platforms may offer a uniquely interactive remote educational experience in surgical training. Strategies and suggestions for its use are discussed, as well as broader considerations in using technology in surgical education.


Coptidis Rhizoma: a comprehensive review of its traditional uses, botany, phytochemistry, pharmacology and toxicology.
Coptidis rhizome (CR), also known as Huanglian in Chinese, is the rhizome of Coptis chinensis Franch., C. deltoidea C.Y. Cheng et Hsiao, or C. teeta Wall (Ranunculaceae). It has been widely used to treat bacillary dysentery, diabetes, pertussis, sore throat, aphtha, and eczema in China. The present paper reviews the latest advances of CR, focusing on the botany, phytochemistry, traditional usages, pharmacokinetics, pharmacology and toxicology of CR and its future perspectives. Studies from 1985 to 2018 were reviewed from books; PhD. and MSc. dissertations; the state and local drug standards; PubMed; CNKI; Scopus; the Web of Science; and Google Scholar using the keywords Coptis, Coptidis Rhizoma, Huanglian, and goldthread. Currently, 128 chemical constituents have been isolated and identified from CR. Alkaloids are the characteristic components, together with organic acids, coumarins, phenylpropanoids and quinones. The extracts/compounds isolated from CR cover a wide pharmacological spectrum, including antibacterial, antivirus, antifungal, antidiabetic, anticancer and cardioprotective effects. Berberine is the most important active constituent and the primary toxic component of CR. As an important herbal medicine in Chinese medicine, CR has the potential to treat various diseases. However, further research should be undertaken to investigate the clinical effects, toxic constituents, target organs and pharmacokinetics, and to establish criteria for quality control, for CR and its related medications. In addition, the active constituents, other than alkaloids, in both raw and processed products of CR should be investigated.


Role of the carboxy-terminal residues of the alpha-subunit in the expression and bioactivity of human thyroid-stimulating hormone.
The glycoprotein hormones TSH, CG, LH, and FSH are heterodimers consisting of a hormone-specific beta-subunit and a common alpha-subunit. The aim of the present study was to investigate the role of the carboxy terminus of the common alpha-subunit (amino acids Tyr89-His90-Lys91-Ser92), which has been shown to be important for human (h) CG and hFSH, for the activity of hTSH. Successive truncations of the alpha-carboxy terminus by site-directed mutagenesis revealed a stepwise reduction of bioactivity occurring at residues alpha Ser92 and alpha His90 to 64% and 13%, respectively. This contrasts with previous findings for hCG and hFSH, where loss of bioactivity occurred in a single step with the deletion of alpha Lys91 but alpha Ser92 was not important. The decreased bioactivities of the hTSH alpha-truncation mutants were reflected by concomitant reductions of cAMP production, thyroid hormone synthesis and cell growth and were accompanied by a loss of receptor binding. Substitution of residues alpha Lys91 or alpha His90 with either a hydrophobic or a bulkier residues resulted in a reduction of receptor binding and signal transduction, indicating that the alpha-carboxy terminus of hTSH may interact with the TSH receptor in a tight contact area. Conversely, substitution of alpha His90 with smaller residues enhanced bioactivity. In addition, the integrity of the alpha-carboxy terminus was essential for hTSH expression. Thus, we showed common and different roles of the alpha-carboxy-terminal residues for the glycoprotein hormones. The unique role of alpha Ser92 in hTSH activity explains the evolutionary constraint to preserve the alpha-carboxy-terminal Ser92 in all glycoprotein hormones.


A mechanism for selective induction of 2'-5' oligoadenylate synthetase, anti-viral state, but not MHC class I genes by interferon-beta in neurons.
Interferon-inducible expression of major histocompatibility class I genes has previously been found to be quantitatively and functionally deficient in neurons compared to other somatic cells or other neural cell types including astrocytes. This deficiency is a key component of neuronal immunoprivilege during viral infections of the CNS. To the contrary, in the present study, induction of functional antiviral state by IFN-beta in neurons compared to astrocytes was found to be highly efficient with respect to both viral replication and protection from cytopathic effects. A candidate antiviral state gene found to be efficiently induced in neurons by IFN-beta was the 2'-5'-oligoadenylate synthetase (OAS) gene. Unlike MHC class I genes, induction of OAS was comparable in neurons and astrocytes indicating differential expression in these neural cell types. Analysis of OAS gene promoter activity indicated that induction of the OAS gene by IFN-beta was dependent on a region containing the interferon stimulated responsive element (ISRE). In contrast, a construct containing the MHC class I-ISRE responsible for induction by IFN-beta in astrocytes was not responsive to IFN-beta in neurons. Therefore, transcription factor binding to the OAS- and MHC-ISREs was analyzed. While the OAS and MHC Class I site bound equal amounts of the transcriptional repressor IRF-2, the OAS-ISRE preferentially interacted with the transcriptional activator ISGF3 in response to IFN-beta. Further, unlike neurons, upregulation of MHC class I genes in astrocytes was related to binding of IRF-1 instead of IRF-2 to the MHC-ISRE. It is proposed that selective activation of anti-viral state genes compared to MHC class I genes by IFN-beta in neurons is mediated by preferential induction and binding of ISGF3 to anti-viral state gene ISREs but not the MHC-ISRE.


Dual Effects of Korean Red Ginseng on Astrocytes and Neural Stem Cells in Traumatic Brain Injury: The HO-1-Tom20 Axis as a Putative Target for Mitochondrial Function.
Astrocytes display regenerative potential in pathophysiologic conditions. In our previous study, heme oxygenase-1 (HO-1) promoted astrocytic mitochondrial functions in mice via the peroxisome-proliferator-activating receptor-γ coactivator-1α (PGC-1α) pathway on administering Korean red ginseng extract (KRGE) after traumatic brain injury (TBI). In this study, KRGE promoted astrocytic mitochondrial functions, assessed with oxygen consumption and adenosine triphosphate (ATP) production, which could be regulated by the translocase of the outer membrane of mitochondria 20 (Tom20) pathway with a PGC-1α-independent pathway. The HO-1-Tom20 axis induced an increase in mitochondrial functions, detected with cytochrome c oxidase subunit 2 and cytochrome c. HO-1 crosstalk with nicotinamide phosphoribosyltransferase was concomitant with the upregulated nicotinamide adenine dinucleotide (NAD)/NADH ratio, thereby upregulating NAD-dependent class I sirtuins. In adult neural stem cells (NSCs), KRGE-treated, astrocyte-conditioned media increased oxygen consumption and Tom20 levels through astrocyte-derived HO-1. HO inactivation by Sn(IV) protoporphyrin IX dichloride in TBI mice administered KRGE decreased neuronal markers, together with Tom20. Thus, astrocytic HO-1 induced astrocytic mitochondrial functions. HO-1-related, astrocyte-derived factors may also induce neuronal differentiation and mitochondrial functions of adult NSCs after TBI. KRGE-mediated astrocytic HO-1 induction may have a key role in repairing neurovascular function post-TBI in peri-injured regions by boosting astrocytic and NSC mitochondrial functions.


Synthesis and Functional Characterization of Novel Sialyl LewisX Mimic-Decorated Liposomes for E-selectin-Mediated Targeting to Inflamed Endothelial Cells.
Sialyl LewisX (sLeX) is a natural ligand of E-selectin that is overexpressed by inflamed and tumor endothelium. Although sLeX is a potential ligand for drug targeting, synthesis of the tetrasaccharide is complicated with many reaction steps. In this study, structurally simplified novel sLeX analogues were designed and linked with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol-2000 (DSPE-PEG) for E-selectin-mediated liposomal delivery. The sLeX structural simplification strategies include (1) replacement of the Gal-GlcNAc disaccharide unit with lactose to reduce many initial steps and (2) substitution of neuraminic acid with a negatively charged group, i.e., 3'-sulfo, 3'-carboxymethyl (3'-CM), or 3'-(1-carboxy)ethyl (3'-CE). While all the liposomes developed were similar in particle size and charge, the 3'-CE sLeX mimic liposome demonstrated the highest uptake in inflammatory cytokine-treated human umbilical vein endothelial cells (HUVECs), being even more potent than native sLeX-decorated liposomes. Inhibition studies using antiselectin antibodies revealed that their uptake was mediated primarily by overexpressed E-selectin on inflamed HUVECs. Molecular dynamics simulations were performed to gain mechanistic insight into the E-selectin binding differences among native and mimic sLeX. The terminally branched methyl group of the 3'-CE sLeX mimic oriented and faced the bulk hydrophilic solution during E-selectin binding. Since this state is entropically unfavorable, the 3'-CE sLeX mimic molecule might be pushed toward the binding pocket of E-selectin by a hydrophobic effect, leading to a higher probability of hydrogen-bond formation than native sLeX and the 3'-CM sLeX mimic. This corresponded with the fact that the 3'-CE sLeX mimic liposome exhibited much greater uptake than the 3'-CM sLeX mimic liposome.


Hypothesis testing as an approach to the analysis of complex tachycardias--an illustrative case of a preexcitation variant.
The correct elucidation of the electrophysiological substrate and mechanism(s) responsible for a complex arrhythmia requires a systematic approach to the analysis of the electrophysiological data. One approach calls for the formulation of a set of hypotheses that could explain the data obtained during the study. The hypotheses are then tested for compatibility with phenomena observed and the one that agrees with the majority of the findings would represent the most tenable explanation. We present the case of a young girl with a wide QRS complex tachycardia and a history of ventricular preexcitation that illustrates this approach. The complexities were resolved only after intraoperative analysis and surgical ablation of a right-sided accessory pathway with decremental properties, and provides further insight into our understanding of the nodoventricular Mahaim fiber.


ADRA2B gene insertion/deletion polymorphism and artery compliance.
The ADRA2B gene insertion/deletion (I/D) polymorphism is associated with various cardiovascular and metabolic phenotypes. Large (C1) and small (C2) artery compliance, assessed by pulse wave analysis, is considered as sensitive markers or risk factors for cardiovascular disease. Therefore whether the ADRA2B I/D polymorphism is associated with C1 and C2 need to be investigated. A total of 227 men and 243 women were enrolled in a Chinese family-based study. C1 and C2 were measured by pulse wave analysis. ADRA2B genotypes were determined by polymerase chain reaction. Statistical methods included generalized estimation equations and quantitative transmission disequilibrium test. The II (31.9%), ID (46.8%) and DD (21.3%) genotype frequencies were in Hardy-Weinberg equilibrium (P = 0.73). The covariates selected by stepwise regression for C1 and C2 were age, systolic pressure and gender. The population based association analysis showed that C1 and C2 were not associated with ADRA2B genotype both before (C1: P = 0.28; C2: P = 0.27) and after (C1: P = 0.58; C2: P = 0.18) the adjustment. The family-based analyses of 128 informative offspring showed that transmission of the D-allele was not associated with C1 or C2, both before (C1: P = 0.42; C2: P = 0.85) and after (C1: P = 0.31; C2: P = 0.82) the adjustment. The study do not support that the ADRA2B gene I/D polymorphism has a major gene effect on C1 or C2 in the Chinese population of current sample size.


Search and clustering orders of magnitude faster than BLAST.
Biological sequence data is accumulating rapidly, motivating the development of improved high-throughput methods for sequence classification. UBLAST and USEARCH are new algorithms enabling sensitive local and global search of large sequence databases at exceptionally high speeds. They are often orders of magnitude faster than BLAST in practical applications, though sensitivity to distant protein relationships is lower. UCLUST is a new clustering method that exploits USEARCH to assign sequences to clusters. UCLUST offers several advantages over the widely used program CD-HIT, including higher speed, lower memory use, improved sensitivity, clustering at lower identities and classification of much larger datasets. Binaries are available at no charge for non-commercial use at http://www.drive5.com/usearch.


A cyclooxygenase-2 (COX-2) selective non-steroidal anti-inflammatory drug enhances the growth inhibitory effect of butyrate in colorectal carcinoma cells expressing COX-2 protein: regulation of COX-2 by butyrate.
Epidemiological, clinical, animal and laboratory studies have all provided evidence for the protective effects of non-steroidal anti-inflammatory drugs (NSAIDs), such as aspirin, against colorectal cancer. The main established target for NSAID action is cyclooxygenase (COX) and the inducible isoform, COX-2, is up-regulated in colorectal cancer. Rat intestinal epithelial cells transfected with a COX-2 expression vector have previously been found to be resistant to butyrate-induced apoptosis. Butyrate, a by-product of dietary fibre fermentation, is known to induce differentiation and apoptosis in colorectal tumour cells in vitro. In recent years there has been considerable interest in the possible role of dietary fibre/resistant starch in the prevention of colorectal cancer. In this study we investigated whether inhibition of COX-2 with a highly selective COX-2 inhibitor (NS-398) would sensitize human colorectal carcinoma cells to the growth inhibitory effect of butyrate. HT29 and S/KS colorectal carcinoma cell lines were treated for 72 h with 2 mM butyrate and/or 10 microM NS-398. Addition of 10 microM NS-398 alone (to inhibit COX-2 activity) did not result in detectable growth inhibition in either of the cell lines. NS-398 enhanced sensitivity to the growth inhibitory effect of butyrate in HT29 cells expressing COX-2 protein. In contrast, NS-398 did not sensitize S/KS cells lacking detectable COX-2 protein and function (as determined by prostaglandin E(2) production) to the growth inhibitory effect of butyrate. In addition, we report that butyrate treatment of carcinoma (HT29) and adenoma (PC/AA/C1) cells leads to up-regulation of COX-2 protein. Thus NS-398 only appears to sensitize human colorectal carcinoma cells expressing COX-2 protein to the growth inhibitory effect of butyrate. As COX-2 is up-regulated in colorectal carcinogenesis, this could have important implications for the selective inhibition of cells expressing COX-2 protein over those lacking COX-2 protein expression and for dietary modification to be considered alongside NSAIDs in the prevention, and possibly treatment, of colorectal cancer.


Diaqua-bis-[5-(pyrazin-2-yl-κN)-3-(pyridin-3-yl)-1,2,4-triazolido-κN]cadmium.
In the title compound, [Cd(C(11)H(7)N(6))(2)(H(2)O)(2)], the Cd(II) cation is located on an inversion center and is coordinated by four N atoms from two 5-(pyrazin-2-yl)-3-(pyridin-3-yl)-1,2,4-triazol-ide anions and two water mol-ecules in a distorted octa-hedral geometry. The triazolide ligand is nearly planar: the central triazole ring is oriented at dihedral angles of 4.63 (13) and 8.41 (13)° with respect to the pyrazine and pyridine rings. Inter-molecular O-H⋯N hydrogen bonds link the mol-ecules into a two-dimensional supra-molecular network parallel to (001).


Fluorescence Phasor Plots Using Time Domain Data: Effect of the Instrument Response Function.
Phasor plots of the fluorescence intensity decay (plots of the Fourier sine transform vs the Fourier cosine transform, for one or several angular frequencies) are being increasingly used, namely, in fluorescence lifetime imaging microscopy (FLIM) of cells, tissues, and surfaces, but are also relevant for the characterization of homogeneous (e.g., solution) systems. In this work, the construction of the phasor plot using time domain data is discussed, including the effect of the instrument response function (IRF). A deconvolution method in the Fourier space is described. The results obtained are applied to fluorescence decays of aqueous fluorescein (basic form) in the presence of concentrated potassium iodide. The effect of the impulse is clearly shown, in accordance with model predictions. Deconvolution in the Fourier space works well for lifetimes at least 1 order of magnitude higher than the IRF time width.


Generation of a membrane potential by electron transport in plasmalemma-enriched vesicles of cotton and radish.
Plasmalemma-enriched vesicles were isolated from cotton roots (Gossypium hirsutum L. cv Acala San Jose 2) and from germinating radish seeds (Raphanus sativa L. cv Tondo Rosso Quarantino). When 100 millimolar ascorbate was added to the grinding medium, the addition of ferricyanide to either preparation led to an inside positive membrane potential as measured by the accumulation of thiocyanate. It is suggested that electrons from ascorbate were being transported electrogenically across the membrane to ferricyanide, resulting in an accumulation of protons within the vesicle. The redox activity of the vesicles has some similarities to that occurring in intact cells, thus providing a simpler system to study the components and effects of transmembrane electron transport.


The total antioxidant power of semen and its correlation with the fertility potential of human male subjects.
There are growing evidences that the damage which is caused to the spermatozoa by the Reactive Oxygen Species (ROS) plays a key role in the male infertility. The seminal plasma is endowed with many enzymatic and nonenzymatic antioxidants which protect the spermatozoa against oxidative stress.The present study was undertaken by using a simple, colourimetric, ferric reducing, antioxidant power for assessing the total antioxidant power rather than the individual antioxidants. The measurement of the individual antioxidants in the seminal plasma, such as Superoxide Dismutase, Vitamin E, etc. is time consuming, which often requires sophisticated and expensive techniques and these measurements may not correlate with the quality of semen. To evaluate the total antioxidant capacity of seminal plasma by estimating the Ferric Reducing Antioxidant Power (FRAP) of semen in different groups of subjects and to correlate it with the different seminogram parameters. The semen samples were obtained from 150 male partners of infertile couples who attended the Reproductive Biology Unit (Infertility Clinic) of the Department of Physiology, MGIMS, Sevagram, who were aged 20-58 years and they were analyzed for the routine seminogram parameters. All the subjects were categorized into two main groups, A. The subjects with abnormal ejaculates, who were further sub classified into the following groups i) Asthenoteratozoospermics (n=25) ii) Oligoasthenoteratozoospermics (n=26) and iii) Azoospermics (n=19) and B. The subjects with normal ejaculates (n=80). The total antioxidant power was measured spectrophotometrically by using the Ferric Reducing Antioxidant Power (FRAP) assay. The Total Antioxidant Capacity (TAC) was found to be significantly lower in the abnormal ejaculates than in the normal ejaculates. A statistically significant positive correlation was observed between the TAC and all the seminogram parameters such as the sperm concentration, sperm motility and sperm morphology (p<0.05). A decreased seminal plasma antioxidant capacity (TAC) could have significant role in the aetiology of impaired sperm functions. So, the tac may be used as specific biomarker for assessing the oxidative stress in sperms.


[Pulmonary biopsy by thoracoscopy: techniques and results in 9 patients].
Video-assisted endoscopic techniques have decreased the surgical aggression in abdominal and thoracic surgery. In our country, pediatric laparoscopy has been developing slowly, but this is not the case for thoracoscopy. The aim of this paper is to present the techniques and results of thoracoscopy pulmonary biopsy in our first patients. Pulmonary biopsies with this approach had been done in 9 patients (5 males, 3 females). Their age ranged between 30 months and 16 years. In all cases this was the last resort for the diagnosis of pulmonary condensation of unknown etiology. The biopsies were done with the pretied knot in 5 cases, stappler in 1 case and with biopsy forceps in 2 cases. Thoracotomy was necessary in one patient, due to intraoperative haemorrhage. Enough tissue for bacterial and pathological diagnosis was obtained. There was not mortality nor important morbidity related with the technique. Postoperative recovery is better when compared with conventional thoracotomy. Thoracoscopy is an adequate approach to perform pulmonary biopsies in children. The advantages if we compare with open thoracotomy are: 1. The possibility to choose the are to perform a minimally invasive biopsy. 2. To take samples of different pulmonary lobes. 3. Less postoperative pain and shorter hospital stay (36-48 hours).


Clinical use of immunosuppressants in Duchenne muscular dystrophy.
Duchenne muscular dystrophy (DMD) is a degenerative disease primarily affecting voluntary muscles with secondary consequences on heart and breathing muscles. DMD is an X-linked recessive disease that results in the loss of dystrophin, a key muscle protein. Inflammation can play different roles in DMD; it can be a secondary response to muscle degeneration, a primary cause of degeneration, or can contribute to the disease progression. Several immunosuppressants have been used with the aim to reduce the inflammation associated with DMD. Most recently, myoblast transplantation has shown the possibility to restore the dystrophin lack in the DMD patient's muscle fibers and this evidence has emphasized the importance of the use of immunosuppressants and the necessity of studying them and their secondary effects. The aim of this review is to analyze the main immunosuppressants drugs starting from the mdx mice experiments and concluding with the most recent human clinical studies.


[Determination of angiotensin-converting enzyme activity in internal diseases].
A convenient and reliable colorimetric method for determination of serum angiotensin-converting enzyme is worked out by modifying the method of Niels and al. Reference serum ranges of 15.45-191.2 ME/L are established in a control group of 46 healthy persons. In pulmonary sarcoidosis statistically significantly higher angiotensin-converting enzyme activity than in the healthy controls is found while in pulmonary cancer and other pulmonary diseases the angiotensin-converting enzyme activity is in the reference ranges. The importance of angiotensin-converting enzyme as a laboratory index for the early diagnosis of pulmonary sarcoidosis is discussed.


Mechanism of potentiation of a dysfunctional epilepsy-linked mutated GABA(A) receptor by a neurosteroid (3alpha, 21-dihydroxy-5alpha-pregnan-20-one): transient kinetic investigations.
The malfunction of a mutated GABA(A) receptor (alpha1beta2gamma2L(K289M)) in an inheritable form of epilepsy (GEFS+, generalized epilepsy with febrile seizures plus) in humans [Baulac, S., Huberfeld, G., Gourfinkel-An, I., Mitropoulou, G., Beranger, A., Prud'homme, J. F., Baulac, M., Brice, A., Bruzzone, R., and LeGuern, E. (2001) Nat. Genet. 28, 46-48] has been accounted for by a 5-fold decrease in the channel-opening equilibrium of the mutated receptor compared to the wild type [Ramakrishnan, L., and Hess, G. P. (2004) Biochemistry 43, 7534-7540]. Here we describe the mechanism by which the neurosteroid 3alpha, 21-dihydroxy-5alpha-pregnan-20-one (5alpha-THDOC) alleviates this malfunction of the mutated receptor transiently expressed in HEK293 cells. Two rapid reaction techniques, the cell-flow and the laser-pulse photolysis methods, were used in combination with whole-cell current recordings. 150-muM 5alpha-THDOC does not affect the rate constant for channel opening (k(op)) of approximately 250 s(-1) but does decrease the rate constant for channel closing (k(cl)) from 121 +/- 11 s(-1) to 56 +/- 21 s(-1). This results in an increase in the channel-opening equilibrium constant ((Phi(-1) = k(op)/k(cl)) by a factor of about 2, leading to about 50% alleviation of the malfunction of the inheritable mutated (alpha1beta2gamma2L(K289M)) GABA(A) receptor linked to GEFS+.


Photosynthesis, Ribulose-1,5-bisphosphate Carboxylase, Electron Transport, and Ribulose 1,5-Bisphosphate of Virescent and Normal Green Wheat Leaves.
CO(2) gas exchange, ribulose-1,5-bisphosphate, and electron transport have been measured in leaves of a yellow-green mutant of wheat (Triticum durum var Cappelli) and its wild type strain grown in the field. All these parameters, expressed on leaf area basis, were similar in both genotypes except electron transport which was more than double in the wild type. These results, treated according to a recent photosynthesis model for C(3) plants, seem to indicate that the electron transport rate of mutant leaves is not sufficient to support the carboxylation derived through both the assimilation rate and the in vitro ribulose-1,5-bisphosphate carboxylase activity. It is suggested that under our experimental conditions photosynthetic electron transport is not the sole energy-dependent determinant of ribulose-1,5-bisphosphate regeneration in the mutant.


Biomechanics of the Human Middle Ear with Viscoelasticity of the Maxwell and the Kelvin-Voigt Type and Relaxation Effect.
The middle ear is one of the smallest biomechanical systems in the human body and is responsible for the hearing process. Hearing is modelled in different ways and by various methods. In this paper, three-degree-of-freedom models of the human middle ear with different viscoelastic properties are proposed. Model 1 uses the Maxwell type viscoelasticity, Model 2 is based on the Kelvin-Voigt viscoelasticity, and Model 3 uses the Kelvin-Voigt viscoelasticity with relaxation effect. The primary aim of the study is to compare the models and their dynamic responses to a voice excitation. The novelty of this study lies in using different models of viscoelasticity and relaxation effect that has been previously unstudied. First, mathematical models of the middle ear were built, then they were solved numerically by the Runge-Kutta procedure and finally, numerical results were compared with those obtained from experiments carried out on the temporal bone with the Laser Doppler Vibrometer. The models exhibit differences in the natural frequency and amplitudes near the second resonance. All analysed models can be used for modelling the rapidly changing processes that occur in the ear and to control active middle ear implants.


Role of implantable cardioverter defibrillator therapy in the management of high-risk patients.
Cardiovascular mortality from ventricular tachycardia (VT) and ventricular fibrillation (VF) continues to be a major health problem. Several therapeutic approaches are now available to treat patients with known VT/VF. Among the various therapeutic options are antiarrhythmic drugs, catheter or surgical ablation of VT focus, and implantable cardioverter defibrillator (ICD). The overall 2-year cardiovascular mortality is significantly reduced by ICD therapy. The ICD is particularly useful in patients with 1) no inducible but clinical VT/VF, 2) drug refractory VT/VF, and 3) VT/VF in association with left ventricular ejection fraction of less than or equal to 30%. Significant improvements in ICD therapy have already been made; these improvements include tiered antitachycardia therapy, antibradycardia pacing, lower defibrillation threshold, and longer life of generator. Further improvements are expected, including nonthoracotomy approach to defibrillation, pectoral implant, and dual chamber sensing. It is likely that with all of the advances in ICD therapy its acceptance as a therapeutic option will increase.


COVID-19, Food Insecurity, and Migration.
In this policy piece, we investigate the coronavirus disease 2019 (COVID-19)-food-insecurity migration channel and develop a policy agenda. The interaction between COVID-19 and the drop in economic activity will lead to increased food insecurity within and across countries. Higher food insecurity may act as a multiplier for the epidemic due to its negative health effects and increased migration. Research has shown that food insecurity affects within-country and cross-border migration. Besides the mean prevalence rate, the distribution of food insecurity affects the migration decision. The impacts of COVID-19 are particularly strong for people in the lower tail of the food-insecurity distribution. In the current context, the effect of food insecurity therefore could be increased migration, including both rural-urban migration and international migration. Importantly, the crisis might lead to a structural break in migration patterns. People might avoid heavily affected COVID-19 destination countries (e.g., United States, Italy, or Spain) and move to other countries. Due to the persistent nature of migration flows, this could have long-lasting effects.


Application of geostatistics with Indicator Kriging for analyzing spatial variability of groundwater arsenic concentrations in Southwest Bangladesh.
This article seeks to explore the spatial variability of groundwater arsenic (As) concentrations in Southwestern Bangladesh. Facts about spatial pattern of As are important to understand the complex processes of As concentrations and its spatial predictions in the unsampled areas of the study site. The relevant As data for this study were collected from Southwest Bangladesh and were analyzed with Flow Injection Hydride Generation Atomic Absorption Spectrometry (FI-HG-AAS). A geostatistical analysis with Indicator Kriging (IK) was employed to investigate the regionalized variation of As concentration. The IK prediction map shows a highly uneven spatial pattern of arsenic concentrations. The safe zones are mainly concentrated in the north, central and south part of the study area in a scattered manner, while the contamination zones are found to be concentrated in the west and northeast parts of the study area. The southwest part of the study area is contaminated with a highly irregular pattern. A Generalized Linear Model (GLM) was also used to investigate the relationship between As concentrations and aquifer depths. A negligible negative correlation between aquifer depth and arsenic concentrations was found in the study area. The fitted value with 95 % confidence interval shows a decreasing tendency of arsenic concentrations with the increase of aquifer depth. The adjusted mean smoothed lowess curve with a bandwidth of 0.8 shows an increasing trend of arsenic concentration up to a depth of 75 m, with some erratic fluctuations and regional variations at the depth between 30 m and 60 m. The borehole lithology was considered to analyze and map the pattern of As variability with aquifer depths. The study has performed an investigation of spatial pattern and variation of As concentrations.


Method for the determination of vitamin K1(20) in human plasma by stable isotope dilution/gas chromatography/mass spectrometry.
A method for the quantitative determination of vitamin K1(20) (VK), an essential cofactor in the carboxylation of clotting factors, is presented. The assay is based on gas chromatography/electron impact mass/spectrometry. The preparation of deuterium-labelled vitamin K1(20) for use as an internal standard is described. The method involves extraction of VK from human plasma and its derivatization to the heptafluorobutyryl ester after reduction of one carbonyl group with zinc. The detection limit was found to be 1.0 pg and the limit of quantitation 2.0 pg ml-1 plasma. This permits the measurement of vitamin K1(20) even in small quantities of plasma, which is highly desirable in investigations dealing with clotting abnormalities in neonates and infants.


Factors Affecting Anxiety and Depression Symptoms in Relatives of Intensive Care Unit Patients.
To determine the incidences of anxiety and depression in relatives of patients admitted to an intensive care unit (ICU) and to investigate the relationships between psychological symptoms and demographic features of the patients and their relatives. Relatives of 78 ICU patients were enrolled in the study. Sociodemographic features of patients and their relatives were recorded. The Turkish version of the Hospital Anxiety and Depression Scale was used to assess anxiety and depression. Twenty-eight (35.9%) cases with anxiety and 56 (71.8%) cases with depression were identified. The mean anxiety and depression scores were 9.49 ± 4.183 and 9.40 ± 4.286, respectively. Anxiety (P = .028) and concomitant anxiety with depression (P = .035) were more frequent among family members of young patients. The relationship to the patient, especially being a spouse, was significantly associated with symptoms (anxiety, P = .009; depression, P = .019; and both, P = .005). Spouses and family members of relatively young patients had higher rates of anxiety and depression. In contrast to the literature, depression was more common than anxiety among the relatives of ICU patients. Further research is needed on the impact of cultural and regional differences on anxiety and depression rates in family members of ICU patients.


A new proposal for radical hysterectomy.
The authors revised the surgical procedure for radical hysterectomy utilizing detailed observation of the venous system, connective fascial sheets, and neural pathways within the uterine supports. The anterior, middle, and posterior uterine supports were reclassified into two systems, supporting or fascial and drainage or areoral. Supporting system consisted of the superficial layer of the vesicouterine ligament, fascial part of the Mackenrodt ligament, sacrouterine ligament, and rectovaginal ligament, whereas drainage system consisted of the deep layer of the vesicouterine ligament, vascular part of the Mackenrodt ligament, and the so-called mesoureter. The operative procedure was planned according to the continuity of these ligaments and executed first by excising the fascia and then dissecting the denuded areoral tissue. Among the 15 patients who underwent surgery for uterine cancer during a 2-year period the mean (SD) time required for the operation was 305.5 +/- 30.5 min and the mean (SD) total volume of blood loss 592.0 +/- 238.2 ml. A mean (SD) period of 14.3 +/- 3.8 postoperative days was required until the volume of the residual urine decreased to less than 50 mL. The present operation has been structured more three-dimensionally and systematically than before. Further, safety of the operation was significantly improved including prevention of hemorrhage and preservation of bladder function.


Modified Endoscopic Vacuum Therapy for Nonhealing Esophageal Anastomotic Leak: Technique Description and Review of Literature.
Endoscopic vacuum-assisted closure (EVAC) therapy is increasingly being used as a new promising method for repairing upper gastrointestinal defects of different etiologies with high success rates. EVAC therapy consists of placing a sponge either within the lumen or within an abscess cavity connected with a nasogastric (NG) tube to a negative pressure system, thus decreasing bacterial contamination and edema and promoting granulation tissue proliferation, thereby gradually decreasing the cavity size until complete closure. Herein, we describe a modified technique for EVAC therapy in which the NG tube is passed into the esophagus through an existing intrapleural drain tract using a rendezvous technique. The small residual fistula was amendable to fibrin glue embolization. This allows easier sponge placement and exchange compared to traditional EVAC technique, and allows oral intake during treatment. We also review the literature regarding other endoscopic treatment options for esophageal anastomotic leaks and perforations. The PubMed database was searched using the terms "esophagus," "esophageal," "leak," "fistula," "endoluminal vacuum-assisted closure (VAC)," "endoscopic VAC," "stent," "sealant," "glue," and "over-the-scope clip (OTSC)." Reference lists of identified articles were searched for further articles, and the "similar articles" function was used on all included articles. Complete closure of the nonhealing fistula was achieved after 8 days of EVAC treatment and fibrin glue embolization. Modified EVAC technique as described is feasible and safe. To the best of our knowledge, this is the first description of this technique. The technique allows easier sponge placement and exchange compared to traditional EVAC technique, and allows oral intake during treatment.


ACTH stimulates fructose 2,6-bisphosphate synthesis and glycolysis in Y-1 adrenal tumor cells.
The effect of ACTH on glycolysis has been studied in Y-1 tumor adrenal cells. ACTH caused a sustained increase in the liberation of lactate as well as a stimulation of both basal and glucose-induced fructose 2,6-bisphosphate content. ACTH produces changes also in the activities of phosphofructokinase-1 and phosphofructokinase-2. The addition of Ca2+ or dibutyryl cyclic AMP did not modify neither lactate production nor fructose 2,6-bisphosphate levels. The results suggest that fructose 2,6-bisphosphate regulates ACTH-induced glycolysis at the phosphofructokinase-1 step, although the biochemical mechanism of phosphofructokinase-2 activation remains elusive.


High-Performance Regular Perovskite Solar Cells Employing Low-Cost Poly(ethylenedioxythiophene) as a Hole-Transporting Material.
Herein, we successfully applied a facile in-situ solid-state synthesis of conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) as a HTM, directly on top of the perovskite layer, in conventional mesoscopic perovskite solar cells (PSCs) (n-i-p structure). The fabrication of the PEDOT film only involved a very simple in-situ solid-state polymerisation step from a monomer 2,5-dibromo-3,4-ethylenedioxythiophene (DBEDOT) made from a commercially available and cheap starting material. The ultraviolet photoelectron spectroscopy (UPS) demonstrated that the as-prepared PEDOT film possesses the highest occupied molecular orbital (HOMO) energy level of -5.5 eV, which facilitates an effective hole extraction from the perovskite absorber as confirmed by the photoluminescence measurements. Optimised PSC devices employing this polymeric HTM in combination with a low-cost vacuum-free carbon cathode (replacing the gold), show an excellent power conversion efficiency (PCE) of 17.0% measured at 100 mW cm-2 illumination (AM 1.5G), with an open-circuit voltage (Voc) of 1.05 V, a short-circuit current density (Jsc) of 23.5 mA/cm2 and a fill factor (FF) of 0.69, respectively. The present finding highlights the potential application of PEDOT made from solid-state polymerisation as a HTM for cost-effective and highly efficient PSCs.


Complication rates of different discectomy techniques for the treatment of lumbar disc herniation: a network meta-analysis.
The aim of this network meta-analysis (NMA) was to compare the complication rates of discectomy/microdiscectomy, percutaneous laser disc decompression (PLDD), percutaneous endoscopic lumbar discectomy (PELD), microendoscopic discectomy (MED), and tubular discectomy for symptomatic lumbar disc herniation (LDH). We searched three online databases for randomized controlled trials (RCTs). Overall complication rates, complication rates per general and modified Clavien-Dindo classification schemes, and reoperation rates were considered as primary outcomes. Odds ratio with 95% confidence intervals for direct comparisons and 95% credible intervals for NMA results were reported. Surface under cumulative ranking curve (SUCRA) was used to estimate ranks for each discectomy technique based on the complication rates. In total, 18 RCTs with 2273 patients were included in this study. Our results showed that there was no significant difference in any of the pairwise comparisons. PELD (SUCRA: 0.856) ranked the lowest for overall complication rates. Discectomy/microdiscectomy (SUCRA: 0.599) and PELD (SUCRA: 0.939) ranked the lowest for intraoperative and post-operative complication rates, respectively. Concerning modified Clavien-Dindo classification scheme, PELD (SUCRA: 0.803), MED (SUCRA: 0.730), and PLDD (SUCRA: 0.605) ranked the lowest for the occurrence of type I, II, and III complications, respectively. Tubular discectomy (SUCRA: 0.699) ranked the lowest for reoperation rates. The results of this NMA suggest that discectomy/microdiscectomy and PELD are the safest procedures for LDH with minimal intraoperative and post-operative complications, respectively. PELD, MED, and PLDD are the safest procedures for LDH in terms of minimal rates for complications necessitating conservative, pharmacological, and surgical treatment, respectively. These slides can be retrieved under Electronic Supplementary Material.


The Effect of Functional Knee Braces on Muscular Contributions to Joint Rotational Stiffness in Anterior Cruciate Ligament-Deficient and -Reconstructed Patients.
Functional knee braces are frequently prescribed by physicians to ameliorate the function of individuals with anterior cruciate ligament (ACL) injuries. These braces have been shown in the literature to potentially enhance knee stability by augmenting muscle activation patterns and the timing of muscle response to perturbations. However, very few techniques are available in the literature to quantify how those modifications in lower-limb muscle activity influence stability of the knee. The aim of the present study was to quantify the effect of an off-the-shelf functional knee brace on muscle contributions to knee joint rotational stiffness in ACL-deficient and ACL-reconstructed patients. Kinematic, electromyography, and kinetic data were incorporated into an electromyography-driven model of the lower extremity to calculate individual and total muscle contributions to knee joint rotational stiffness about the flexion-extension axis, for 4 independent variables: leg condition (contralateral uninjured, unbraced ACL injured, and braced ACL injured); knee flexion (5°-10°, 20°-25°, and 30°-35°); squat stability condition (stable and unstable); and injury status (ACL deficient and ACL reconstructed). Participants had significantly higher (P < .05, η2 = .018) total knee joint rotational stiffness values while wearing the brace compared with the control leg. A 2-way interaction effect between stability and knee flexion (P < .05, η2 = .040) for total joint rotational stiffness was also found.


Detection of phosphorylated T and B cell antigen receptor species by Phos-tag SDS- and Blue Native-PAGE.
Detection of phospho-proteins and differently phosphorylated forms of the same protein are important in understanding cell behaviour. One novel method is Phos-tag SDS-PAGE. A dinuclear Mn(2+) complex that binds to phosphate groups (the Phos-tag) is covalently attached to the polyacrylamide gel matrix. Thus, phosphorylated proteins are retarded in their migration and can be distinguished from their non-phosphorylated counterparts. We applied Phos-tag SDS-PAGE to the analysis of the zeta, CD3epsilon and CD3delta subunits of the T cell antigen receptor (TCR-CD3). Pervanadate stimulation generated six different phospho-zeta and each two different CD3epsilon and CD3delta forms. This corresponds to the phosphorylatable tyrosines on their cytoplasmic tails. The phosphorylation pattern was compatible with random phosphorylation events. Further, we showed that the Phos-tag technology can be applied to Blue Native (BN)-PAGE. This extends the applicability to the analysis of native protein complexes. Upon pervanadate stimulation the TCR-CD3 complex was predominantly detected as two distinct phospho-complexes. In contrast, the B cell antigen receptor (BCR) appeared as one phospho-form. Thus, Phos-tag BN-PAGE is useful for the analysis of different phosphorylation states of multiprotein complexes.


Zen practice: a training method to enhance the skills of clinical social workers.
The use of Zen Buddhist meditation in clinical practice has received specific attention from mental health disciplines in the West. A study was undertaken to examine the influence of a personal practice of Zen on the professional work of clinical social workers. Ten experienced clinical social workers who were long-term Zen practitioners were interviewed. Findings from this qualitative study suggest that Zen meditation has direct application to clinical social work in three areas: (1) cultivating Awareness, (2) enhancing Acceptance, and (3) nurturing Responsibility. This article reports on the findings in the area of Awareness. Awareness increases a social worker's focus on the present moment with the client, providing for a suspension of preconceived ideas about the client. Building on the view that social work is both art and science, the author proposes the use of Zen in the training of clinical social workers.


Small genomic insertions form enhancers that misregulate oncogenes.
The non-coding regions of tumour cell genomes harbour a considerable fraction of total DNA sequence variation, but the functional contribution of these variants to tumorigenesis is ill-defined. Among these non-coding variants, somatic insertions are among the least well characterized due to challenges with interpreting short-read DNA sequences. Here, using a combination of Chip-seq to enrich enhancer DNA and a computational approach with multiple DNA alignment procedures, we identify enhancer-associated small insertion variants. Among the 102 tumour cell genomes we analyse, small insertions are frequently observed in enhancer DNA sequences near known oncogenes. Further study of one insertion, somatically acquired in primary leukaemia tumour genomes, reveals that it nucleates formation of an active enhancer that drives expression of the LMO2 oncogene. The approach described here to identify enhancer-associated small insertion variants provides a foundation for further study of these abnormalities across human cancers.


Malignant histiocytoses developing in patients with B-cell lymphomas. Report of two cases.
Two patients with indolent B-cell lymphomas (small B-cell and small cleaved cell) developed a coexistent malignant histiocytosis 3 and 6 years after diagnosis of their lymphomas. In both patients, malignant histiocytosis presented as new onset of fever, weight loss, and rapidly progressive bulky disease super-imposed on a previously stable clinical course without constitutional symptoms. Both patients failed to respond to combination chemotherapy. The terminal aggressive phase of indolent B-cell lymphomas usually represents transformation of the original neoplasm to a large non-cleaved or immunoblastic B-cell histology. Malignant histiocytosis has not previously been reported as a second neoplasm in patients with B-cell lymphomas.


Reconstruction of gene regulatory networks under the finite state linear model.
We study the Finite State Linear Model (FSLM) for modelling gene regulatory networks proposed by A. Brazma and T. Schlitt in [4]. The model incorporates biologically intuitive gene regulatory mechanism similar to that in Boolean networks, and can describe also the continuous changes in protein levels. We consider several theoretical properties of this model; in particular we show that the problem whether a particular gene will reach an active state is algorithmically unsolvable. This imposes some practical difficulties in simulation and reverse engineering of FSLM networks. Nevertheless, our simulation experiments show that sufficiently many of FSLM networks exhibit a regular behaviour and that the model is still quite adequate to describe biological reality. We also propose a comparatively efficient O(2(K)n(K+1)M(2K)m log m) time algorithm for reconstruction of FSLM networks from experimental data. Experiments on reconstruction of random networks are performed to estimate the running time of the algorithm in practice, as well as the number of measurements needed for successful network reconstruction.


[Bridging osteosynthesis in closed compound fractures of the femur shaft].
The technique of bridging plate osteosynthesis in closed comminuted fractures of the femoral shaft is described. With regard to clinical and radiological results as well as to complications in the postoperative course a group of 17 fractures stabilized by the bridging technique is compared with a group of 16 femoral fractures fixed by plate osteosynthesis after anatomical reduction.


Motility and flagellum synthesis in Halobacillus halophilus are chloride dependent.
The motility of Halobacillus halophilus as observed on swarm agar plates was strictly dependent on the chloride concentration. Cl(-) was apparently not used as the coupling ion for flagellar rotation. Cells grown in the absence of chloride were devoid of flagella, but flagellation was restored upon the addition of chloride. These experiments indicate that chloride is involved in synthesis of flagella in H. halophilus.


Selective phosphodiesterase-3 inhibitor cilostazol ameliorates experimental autoimmune encephalomyelitis.
We investigated the possible therapeutic effect of cilostazol, a specific inhibitor of phosphodiesterase-3, for experimental autoimmune encephalomyelitis (EAE). Mice affected with EAE induced by inoculation with MOG(35-55) were fed with cilostazol or vehicle control. The clinical EAE scores of the cilostazol-fed mice were lower than those of the controls. Serum level of soluble intercellular adhesion molecule-1 was significantly lower in the cilostazol-fed mice than in the controls. In the recall responses with MOG(35-55), proliferation and IFN-gamma production by lymphocytes from cilostazol-fed mice were significantly reduced. Cilostazol may exhibit repressive effects on EAE by reducing the antigen-specific T-cell response and decreasing the expression of the adhesion molecules. Cilostazol is a hopeful choice for the treatment of multiple sclerosis.


English community pharmacists' experiences of using electronic transmission of prescriptions: a qualitative study.
The Electronic Prescription Service Release 2 (EPS2) in England has been designed to provide electronic transmission of digitally-signed prescriptions between primary care providers, with the intent on removing the large amounts of paper currently exchanged. As part of a wider evaluation of the EPS service, we wished to explore pharmacists' experience with the new system and their perceptions of its benefits and any associated problems. We conducted semi-structured telephone interviews with community pharmacists using EPS2. We used a purposive sampling technique to obtain views from pharmacists working in pharmacies with a range of sizes and locations and to include both independent pharmacies and multiples. Interviews were transcribed verbatim and coded using grounded theory to identify the main factors that have influenced deployment and implementation in the eyes of respondents. QSR Nvivo was used as to aid in this process. It became apparent from the analysis that respondents perceived a wide range of advantages of EPS including improved safety, stock control, time management and improved relationships between pharmacy and General Practice staff. Respondents did not perceive a large difference in terms of work processes or development of their professional role. A large number of problems had been experienced in relation to both the technology itself and the way it was used by General Practices. It became apparent that work-around procedures had been developed for dealing with these issues but that not all these problems were perceived as having been addressed sufficiently at source. This sometimes had implications for the extent of EPS2 use and also limited some of the potential advantages of the EPS2 system, such as reduced effort in the management of prescription reimbursement. Respondents made suggestions for future improvements to EPS2. While interview data demonstrated that there were some feedback procedures in place, these were not regarded as being sufficient by the majority of respondents. Whilst pharmacists perceived a wide range of benefits of EPS, a large number of problems had been experienced. Despite these difficulties, no pharmacists expressed an overall negative view.


Epidermotropic CD8+ cytotoxic T-cell lymphoma exhibiting a transition from the indolent to the aggressive phase, accompanied by emergence of CD7+ cells and formation of neutrophilic pustules.
A 47-year-old-man presented with rashes on his trunk and limbs, and a diagnosis of parapsoriasis was made. Ten years later, the rashes had progressed gradually to form plaques and tumours. Gene rearrangement studies revealed monoclonality of the T-cell receptor β-chain (TCR-Jβ)1 gene, and results of flow cytometry and immunohistochemical examination confirmed a diagnosis of epidermotropic CD8+ cytotoxic T-cell lymphoma. The clinical course of the disease remained indolent for some time, but about 2 years later, neutrophilic pustules formed on the surface of the skin lesions, and tumours developed in the patient's testes. Using flow cytometry, emergence of CD7+ cells was found. The patient died the following year of respiratory failure due to brain herniation. On postmortem examination, CD8+ tumour cells were found in the brain. This case demonstrates an unusually protracted indolent phase in a patient with cutaneous aggressive epidermotropic CD8+ cytotoxic T-cell lymphoma; its transition into the aggressive phase was accompanied by emergence of CD7+ cells and formation of neutrophilic pustules.


Amorphous Ruthenium-Sulfide with Isolated Catalytic Sites for Pt-Like Electrocatalytic Hydrogen Production Over Whole pH Range.
Electrocatalytic hydrogen evolution reaction (HER) is an efficient way to generate hydrogen fuel for the storage of renewable energy. Currently, the widely used Pt-based catalysts suffer from high costs and limited electrochemical stability; therefore, developing an efficient alternative catalyst is very urgent. Herein, one pot hydrothermal synthesis is reported of amorphous ruthenium-sulfide (RuSx ) nanoparticles (NPs) supported on sulfur-doped graphene oxide (GO). The as-obtained composite serves as a Pt-like HER electrocatalyst. Achieving a current density of -10 mA cm-2 only requires a small overpotential (-31, -46, and -58 mV in acidic, neutral, and alkaline electrolyte, respectively) with high durability. The isolated Ru active site inducing Volmer-Heyrovsky mechanism in the RuSx NPs is demonstrated by the Tafel analysis and X-ray absorption spectroscopy characterization. Theoretical simulation indicates the isolated Ru site exhibits Pt-like Gibbs free energy of hydrogen adsorption (-0.21 eV) therefore generating high intrinsic HER activity. Moreover, the strong bonding between the RuSx and S-GO, as well as pH tolerance of RuSx are believed to contribute to the high stability. This work shows a new insight for amorphous materials and provides alternative opportunities in designing advanced electrocatalysts with low-cost for HER in the hydrogen economy.


Vesicular stomatitis virus enters cells through vesicles incompletely coated with clathrin that depend upon actin for internalization.
Many viruses that enter cells by clathrin-dependent endocytosis are significantly larger than the dimensions of a typical clathrin-coated vesicle. The mechanisms by which viruses co-opt the clathrin machinery for efficient internalization remain uncertain. Here we examined how clathrin-coated vesicles accommodate vesicular stomatitis virus (VSV) during its entry into cells. Using high-resolution imaging of the internalization of single viral particles into cells expressing fluorescent clathrin and adaptor molecules, we show that VSV enters cells through partially clathrin-coated vesicles. We found that on average, virus-containing vesicles contain more clathrin and clathrin adaptor molecules than conventional vesicles, but this increase is insufficient to permit full coating of the vesicle. We further show that virus-containing vesicles depend upon the actin machinery for their internalization. Specifically, we found that components of the actin machinery are recruited to virus-containing vesicles, and chemical inhibition of actin polymerization trapped viral particles in vesicles at the plasma membrane. By analysis of multiple independent virus internalization events, we show that VSV induces the nucleation of clathrin for its uptake, rather than depending upon random capture by formation of a clathrin-coated pit. This work provides new mechanistic insights into the process of virus internalization as well as uptake of unconventional cargo by the clathrin-dependent endocytic machinery.


Relative long-term effects of spironolactone in conjunction with an angiotensin-converting enzyme inhibitor on left ventricular mass and diastolic function in patients with essential hypertension.
It has been reported that treatment with an angiotensin-converting enzyme (ACE) inhibitor is not adequate to suppress aldosterone, and we previously demonstrated that adding spironolactone to an ACE inhibitor may have beneficial effects on left ventricular hypertrophy (LVH) in selected patients with essential hypertension (EH). We have extended our previous short-term study, and addressed the relative long-term clinical effects of spironolactone and an ACE inhibitor in patients with EH who have LVH. Twenty patients with EH and concomitant LVH participated in this study. Subjects were treated with either an ACE inhibitor alone (group 1: 10 patients) or an ACE inhibitor plus spironolactone at the dose of 25 mg (group 2: 10 patients) for 60 weeks. The baseline clinical and echocardiographic characteristics of the two groups were similar. Final values of blood pressure were also similar between the two groups. The LV mass index (LVMI) decreased significantly in both groups, but the extent of reduction was significantly greater in group 2 at 60 weeks. The early peak to atrial peak filling velosities ratio (E/A ratio) was significantly increased to a similar extent in both groups. Serum procollagen type III amino-terminal peptide (PIIINP) was significantly decreased in group 2, but not in group 1. In group 2, there was a statistically significant correlation between the changes in LVMI and PIIINP. In conclusion, adding spironolactone to therapy with an ACE inhibitor for 60 weeks may have beneficial effects in patients with EH and concomitant LVH. Our study strongly suggests the possibility that attenuation of the effects of cardiac aldosterone in patients with EH by treatment with spironolactone and an ACE inhibitor may become a new goal for the prevention and regression of cardiac hypertrophy.


Loneliness and physical function impairment: Perceived health status as an effect modifier in community-dwelling older adults in Ghana.
Although loneliness and physical function impairment (PFI) are common geriatric syndromes and public health issues, little is known about how their associations vary via self-perception of health. We examine how loneliness is associated with PFI, and whether the association is modified by perceived health status. We conducted a cross-sectional analysis of 1201 adults aged ≥ 50 years from the Aging, Health, Psychological Well-being and Health Seeking Behavior Study (AgeHeaPsyWel-HeaSeeB) in Ghana. We assessed loneliness using the three-item short-form of the UCLA Loneliness Scale, and PFI was measured with a seven-item scale on mobility-related deficiencies. Adjusted logistic regressions and moderation analysis evaluated the hypothesized associations. The prevalence of moderate, severe loneliness, and PFI were 37.5%, 17.7%, and 36.1%, respectively. Regressions showed that loneliness was associated with a 23% increased risk of PFI after adjusting for several potential confounders (OR = 1.23; 95%CI = 1.03-2.81). PFI sub-types revealed similar risks. The loneliness-PFI association was significantly moderated by perceived health status such that a positive health perception attenuated the effect of loneliness on PFI (OR = 0.46, 95%CI = 0.23-0.90). Individuals who were lonely had significantly higher odds for PFI but the effect was tempered by perceived health status. Social policy and public health practices for healthy aging should address loneliness and negative health perception among older people.


Outcomes and Toxicities of Nonmedullary Thyroid Tumors Treated with Proton Beam Radiation Therapy.
Proton therapy is an emerging therapy for several malignancies owing to its favorable therapeutic ratio. There are very limited data on the use of proton therapy in the management of thyroid carcinoma. Our objective was to review the safety, feasibility, and outcomes of proton therapy for patients with thyroid cancer treated to the head and neck. From our institution's proton database from 2012 to 2021, we identified 22 patients with thyroid cancer treated with proton beam therapy. We evaluated outcomes and toxicities. Median follow-up was 26 months. Of the 22 patients, 50% were female. The mean age was 65 years. Three patients had anaplastic cancer; 13, papillary carcinoma; 2, follicular carcinoma; and 2, poorly differentiated carcinoma. Forty-six percent had T4 disease. Primary targets were the central neck compartment, level VI, and upper mediastinum. Radiation dose was 60 GyRBE adjuvantly, and 70 GyRBE for gross disease (range, 6000-7600 GyRBE). Eight patients underwent upfront adjuvant radiation, and 3 received definitive radiation for unresectable disease upfront. Eleven patients received either salvage or palliative radiation. Fifty-nine percent of patients had extrathyroidal extension, and 64% of patients had gross disease in the neck before treatment. Fifty percent of patients had metastatic disease before treatment. Sixteen patients received concurrent chemotherapy, 63% of these patients received doxorubicin. For all patients, 1-year local regional recurrence (LRR) was 0%, and overall survival (OS) was 90%. Acute grade 3+ toxicities occurred in 27% of patients, the most frequent being dermatitis (27%). Three patients required a percutaneous endoscopic gastrostomy tube after radiation therapy (RT), 2 owing to progression. There were no grade 4+ toxicities. Proton therapy for thyroid cancer appears feasible and effective with minimal toxicities. Prospective studies comparing proton therapy with intensity-modulated RT, to evaluate the clinical efficacy of using proton therapy to reduce toxicities in patients undergoing radiation for thyroid cancer, are warranted.


First record of the New Guinea flatworm Platydemus manokwari (Platyhelminthes, Geoplanidae) as an alien species in Hong Kong Island, China.
The New Guinea flatworm (Platydemus manokwari) caused extinction of the native land snails on several Pacific island in past decades, and therefore it has been listed among the top 100 of the world's worst invasive alien species. Using morphological and molecular methods, New Guinea flatworms were discovered and identified for the first time in Hong Kong Island during a field investigation in July and August 2018. The flatworms were 32-60 mm long, 3-5 mm wide, and 1-2 mm thick. The dorsal side of the flatworm was dark brown with a thin yellow central line, and its ventral side appeared pale grey. To further verify this species, both 18S rDNA and mitochondrial cytochrome c oxidase subunit I gene (COX1) obtained from three specimens of P. manokwari were sequenced and analysed. While comparing these sequences with those previously deposited in GenBank, these 18S rDNA sequences shared 100% identity with the single available 18S rDNA sequence of P. manokwari; and the obtained COX1 sequences were identical to those of P. manokwari world genotype. Two native snails, Criptosoma imperator and Bradybaena similaris, have been found to be the prey of this predator during this investigation. Therefore, the invasive New Guinea flatworm certainly will cause a serious impact on the biodiversity of native snail populations, and an economic and environmental risk assessment for P. manokwari need to be completed in the near future in Hong Kong.


Model for a one-step plasma treatment device: feasibility of cholesterol removal.
A self-contained plasmapheresis device based on Starling flow has been proposed. A computer model was developed to describe the device performance, and a device using conventional cellulose tri-acetate (CTA) fibers was modeled. Predicted pressure drops and ultrafiltration flow agreed well with measurements using bovine blood in prototype devices made with CTA fibers. The model predicted the expected Starling flow and predicted the observed high hematocrit region near the fiber wall and a layer of packed red cells on the wall. Typically, 15% of the ultrafiltration flow occurred in the first 2-3% of the fiber length where no packed cell layer was present. The entrance header was predicted to cause lower blood pressures in the peripheral fibers, producing a net plasma migration from the central to the peripheral fibers. As an example application for this device, the computer model was used to assess the feasibility of reducing LDL-cholesterol in patients. The use of sequential 1 hour treatments by a modified device was predicted to reduce the cholesterol in a standard man from 200 mg% to 150 mg% in less than 3 hours.


Is the natriuretic effect of atrial natriuretic factor in conscious rats mediated by renal vasodilatation?
Atrial natriuretic factors (ANF) have both vasorelaxant and natriuretic properties. It has been suggested that their effects on sodium excretion depend on renal vasodilatation. We studied the haemodynamic (ultrasound Doppler flow probes) effects of synthetic atriopeptin II at natriuretic doses in conscious rats. Intravenous injections of atriopeptin II induced a marked, dose-dependent increase in sodium excretion. This was accompanied by a slight reduction in blood pressure (BP) and a transient decrease in renal, mesenteric and aortic blood flow. In contrast, a selective renal vasodilator, CGP 22,979, induced a moderate increase in sodium excretion concomitantly with a rise in renal blood flow. Blood pressure and the other regional flows remained unchanged. These findings suggest that the natriuretic effect of CGP 22,979 is closely related to an increased renal blood flow. Conversely, the natriuresis induced by atriopeptin II is not accompanied by selective renal vasodilatation and appears to be due either to a tubular action or to changes in intrarenal haemodynamics.


Treatment of non-small cell lung cancer with intensity-modulated radiation therapy in combination with cetuximab: the NEAR protocol (NCT00115518).
Even today, treatment of Stage III NSCLC still poses a serious challenge. So far, surgical resection is the treatment of choice. Patients whose tumour is not resectable or who are unfit to undergo surgery are usually referred to a combined radio-chemotherapy. However, combined radio-chemotherapeutic treatment is also associated with sometimes marked side effects but has been shown to be more efficient than radiation therapy alone. Nevertheless, there is a significant subset of patients whose overall condition does not permit administration of chemotherapy in a combined-modality treatment. It could be demonstrated though, that NSCLCs often exhibit over-expression of EGF-receptors hence providing an excellent target for the monoclonal EGFR-antagonist cetuximab (Erbitux) which has already been shown to be effective in colorectal as well as head-and-neck tumours with comparatively mild side-effects. The NEAR trial is a prospective phase II feasibility study combining a monoclonal EGF-receptor antibody with loco-regional irradiation in patients with stage III NSCLC. This trial aims at testing the combination's efficacy and rate of development of distant metastases with an accrual of 30 patients. Patients receive weekly infusions of cetuximab (Erbitux) plus loco-regional radiation therapy as intensity-modulated radiation therapy. After conclusion of radiation treatment patients continue to receive weekly cetuximab for 13 more cycles. The primary objective of the NEAR trial is to evaluate toxicities and feasibility of the combined treatment with cetuximab (Erbitux) and IMRT loco-regional irradiation. Secondary objectives are remission rates, 3-year-survival and local/systemic progression-free survival.


A high-throughput and single-tube recombination of crude PCR products using a DNA polymerase inhibitor and type IIS restriction enzyme.
Type IIS restriction enzymes have been successfully used as "universal" restriction enzymes in DNA manipulations. We took a step further to develop a rapid technique for recombining DNA fragments, fully automatic single-tube recombination (FASTR), which enables multiple-fragment DNA recombination in a single step. Crude PCR products are directly mixed with both type IIS restriction endonuclease and DNA ligase to initiate a spontaneous and one-way recombination reaction. Highly efficient DNA recombination can be achieved by an inhibition of DNA polymerase with aphidicolin and a selective digestion of template DNAs by DpnI, a restriction enzyme to digest hemi-methylated DNA in the reaction solution; thereby the entire procedure takes less than 15 min. Owing to its simplicity, efficiency and rapidity, one-step FASTR can be applied to a wide range of DNA manipulations including those involving high-throughput applications where significant reduction in time and cost is expected.


Updates in Grading of Renal Cell Carcinomas Beyond Clear Cell Renal Cell Carcinoma and Papillary Renal Cell Carcinoma.
The World Health Organization (WHO) recommends grading of clear cell renal cell carcinoma (RCC) and papillary RCC using the WHO/International Society of Urological Pathology (ISUP) grade, which is primarily based on nuclear features. As the spectrum of RCC continues to evolve, with more recently described subtypes in the past decade, literature evidence on grading these subtypes is limited or not available for some tumor types. Herein, we outline a pragmatic approach to the topic of grading RCC, dividing the contemporarily described RCC subtypes into 7 categories based on the potential clinical applicability of grading as a useful prognostic parameter: (1) RCC subtypes that are reasonably validated and recommended for WHO/ISUP grading; (2) RCC subtypes where WHO/ISUP is not applicable; (3) RCC subtypes where WHO/ISUP grading is potentially clinically useful; (4) inherently aggressive RCC subtypes where histologic classification itself confers an aggressive biologic potential; (5) renal epithelial tumors where WHO/ISUP grading provides potentially misleading prognostic implication; (6) renal epithelial neoplasms where low WHO/ISUP grade features are a prerequisite for accurate histologic classification; and (7) renal epithelial neoplasms with no or limited data on grading or incomplete understanding of the biologic potential. Our aim in outlining this approach is 2-fold: (a) identify the gaps in understanding and application of grading in RCC subtypes so that researchers in the field may perform additional studies on the basis of which the important pathologic function of assignment of grade may be recommended to be performed as a meaningful exercise across a wider spectrum of RCC; and (b) to provide guidance in the interim to surgical pathologists in terms of providing clinically useful grading information in RCC based on currently available clinicopathologic information.


Competition of electron transfer, dissociation, and bond-forming processes in the reaction of the CO(2)(2+) dication with neutral CO(2).
The bimolecular reactivity of the CO(2)(2+) dication with neutral CO(2) is investigated using triple quadrupole and ion-ion coincidence mass spectrometry. Crucial for product analysis is the use of appropriate isotope labelling in the quadrupole experiments in order to distinguish the different reactive pathways. The main reaction corresponds to single-electron transfer from the neutral reagent to the dication, i.e. CO(2)(2+) + CO(2) --> 2CO(2)(+); this process is exothermic by almost 10 eV, if ground state monocations are formed. Interestingly, the results indicate that the CO(2)(+) ion formed when the dication accepts an electron dissociates far more readily than the CO(2)(+) ion formed from the neutral CO(2) molecule. This differentiation of the two CO(2)(+) products is rationalized by showing that the population of the key dissociative states of the CO(2)(+) monocation will be favoured from the CO(2)(2+) dication rather than from neutral CO(2). In addition, two bond-forming reactions are observed as minor channels, one of which leads to CO(+) and O(2)(+) as ionic products and the other affords a long-lived C(2)O(3)(2+) dication.