Pharmacokinetic and thrombolytic properties of chimeric plasminogen activators consisting of a single-chain Fv fragment of a fibrin-specific antibody fused to single-chain urokinase.
The pharmacokinetic and thrombolytic properties were determined of two recombinant single-chain chimeric plasminogen activators (PA) consisting of u-PA-33k, a low-molecular weight derivative of single-chain urokinase-type PA (scu-PA) comprising amino acids Ala132 through Leu411, and of either a single-chain variable region fragment (Fv) derived from the fibrin fragment D-dimer-specific monoclonal antibody MA-15C5 (K12G0S32) or of the deglycosylated single-chain Fv fragment obtained by substitution of Asn88 with Glu (K12G2S32). Following bolus injection in hamsters, clearances of recombinant scu-PA (rscu-PA) and of K12G0S32 were similar. In contrast, clearance of K12G2S32 was fourfold slower than that of rscu-PA. The thrombolytic potency (percent lysis per u-PA administered in milligrams per kilogram body weight) and specific thrombolytic activity (percent lysis per microgram per milliliter steady-state plasma u-PA antigen level) of these compounds were studied in hamsters with an experimental pulmonary embolus consisting of a human plasma clot injected via the jugular vein. The doses of K12G0S32 and K12G2S32 required to obtain maximal rate of clot lysis were sixfold and 11-fold lower than that of rscu-PA. The steady-state u-PA-related plasma antigen levels of K12G0S32 and K12G2S32 required to obtain maximal rate of clot lysis were 10-fold and fourfold lower than that of rscu-PA. Thus, targeting of K12G0S32 to the clot surface by means of its glycosylated Fv fragment results in a 10-fold increase of its specific thrombolytic activity and sixfold increase of its thrombolytic potency as compared with those of rscu-PA. Targeting of K12G2S32 to the clot surface by means of its deglycosylated Fv fragment results in only a twofold increase of its thrombolytic activity. However, its fourfold slower clearance, combined with its twofold higher specific thrombolytic activity, results in an 11-fold increase of its thrombolytic potency over that of rscu-PA. These findings indicate that the thrombolytic potency of chimeric antibody-targeted PA may be increased by increasing the specific thrombolytic activity, reducing the clearance, or both.


Eotaxin contributes to renal interstitial eosinophilia.
A potent eosinophil chemotactic cytokine, human eotaxin, is directly chemotactic for eosinophils. Therefore, the specific expression of eotaxin in tissue might play a crucial role in tissue eosinophilia. However, the precise molecular mechanism of the recruitment and activation of eosinophils in human renal diseases remains to be investigated. We evaluated the role of eotaxin in the pathogenesis of human diffuse interstitial nephritis with marked infiltration of eosinophils. In this study, we examined 20 healthy volunteers. 56 patients with primary or secondary glomerular diseases and two hypereosinophilic syndrome patients without renal involvement. Urinary and serum eotaxin levels were determined by an enzyme-linked immunosorbent assay. We also detected the presence of eotaxin protein immunohistochemically. On the one hand, urinary levels of eotaxin were significantly higher before the initiation of glucocorticoid administration in the patient with interstitial nephritis with marked infiltration of eosinophils. On the other hand, urinary eotaxin levels were not detected in any patients with nephrotic syndrome, interstitial nephritis without eosinophils, hypereosinophilic syndrome without renal involvement or other renal diseases. Serum eotaxin levels were not detected in any of the patients. Therefore, the detection of eotaxin in the urine was specific for renal interstitial eosinophilia. Moreover, endothelial cells, infiltrating mononuclear cells and renal epithelial cells in the tubulointerstitial lesions were immunostained with specific anti-eotaxin antibodies. Furthermore, the elevated urinary levels of eotaxin decreased dramatically during glucocorticoid-induced convalescence. We hypothesize that in situ expression of eotaxin may provide a new mechanism to explain the renal interstitial eosinophil infiltration.


Sinus rhythm restoration after atrial fibrillation: the clinical value of N-terminal pro-BNP measurements.
To examine the effects of sinus rhythm (SR) restoration on N-Terminal pro-BNP (NTP-BNP) in patients with atrial fibrillation (AF). Subjects with paroxysmal and persistent AF and absence of organic heart disease were prospectively studied. Chemical or electrical restoration of SR was attempted within 48 hours (n = 37) or >3 weeks (n = 73). Clinical and laboratory (NTP-BNP, 72-hour Holter monitor, and electrocardiogram) assessment were obtained at baseline and at 1, 30, and 180 days after SR restoration. Patients were divided into three predefined "outcome groups": (a) maintenance of SR for 1 month, (b) SR with recurrent paroxysmal AF (PaAF), and (c) early (<30 days) recurrence persistent AF (RAF). Of the 110 patients enrolled, 89 had initial successful SR restoration. Baseline NTP-BNP was 936 pg/mL (interquartile range (IQR) 333-2,026); ratio between baseline and 30-day NTP-BNP was 10.2 (IQR 6.42-22.0) for SR group, 3.3 (IQR 2.45-7.34) for PaAF, and 1.07 (IQR 0.87-1.22) for RAF (P < 0.001). Patients with ratio </=3 were more likely to have PaAF (46% vs 3%, OR 30, P < 0.001). With SR restoration, NTP-BNP decline is observed up to 1 month. NTP-BNP drop is partially or completely abolished by PaAF and RAF, respectively. NTP-BNP does not predict successful SR restoration.


Allelic variation of human serotonin transporter gene expression.
Mood, emotion, cognition, and motor functions as well as circadian and neuroendocrine rhythms, including food intake, sleep, and reproductive activity, are modulated by the midbrain raphe serotonin (5-HT) system. By directing the magnitude and duration of postsynaptic responses, carrier-facilitated 5-HT transport into and release from the presynaptic neuron are essential for the fine tuning of serotonergic neurotransmission. Interest in the mechanism of environmental factor-, disease-, and therapy-induced modification of 5-HT transporter (5-HTT) function and its impact on early brain development, event-related synaptic plasticity, and neurodegeneration is widespread and intensifying. We have recently characterized the human and murine 5-HTT genes and performed functional analyses of their 5'-flanking regulatory regions. A tandemly repeated sequence associated with the transcriptional apparatus of the human 5-HTT gene displays a complex secondary structure, represses promoter activity in nonserotonergic neuronal cells, and contains positive regulatory components. We now report a novel polymorphism of this repetitive element and provide evidence for allele-dependent differential 5-HTT promoter activity. Allelic variation in 5-HTT-related functions may play a role in the expression and modulation of complex traits and behavior.


Therapeutic endoscopy: removal of gastrointestinal foreign bodies in children.
Endoscopic removal of foreign bodies (FBs) is an important part of therapeutic endoscopy in children. The majority of FB ingestion occur in children under 5 years. Children most often ingest coins, pins, keys, round stones or marbles, nails, rings, batteries and toys, and coins are the most common in the pediatric series of 139 children, who underwent endoscopic removal in Seoul National University, Children's Hospital. The management of FBs in children depends on the type (sharp or dull, pointed or blunt, and toxic or nontoxic) and the-size, along with the location of FB in the gastrointestinal tract. The endoscopist should decide whether endoscopic intervention is necessary and how urgently it has to be done. Indications for removal of FBs from the gastrointestinal tract in children are 1) all esophageal FBs. 2) gastric and duodenal FBs if they are sharp or pointed, if more than 4 cm long or 2 cm wide in young infants and children, if containing toxic substances and if blunt objects after 2 weeks observation in the stomach or 1 week observation in the duodenum. All FBs impacted in the esophagus should be removed within 24 hours because of the risks of perforation and serious fistula formation. Disk batteries can cause corrosive injury to the esophagus within 4 hours, and should be removed as soon as possible. Coin retrieval can be done very securely by the W-shape FB grasping forcep without an endotracheal general anesthesia. Blunt FBs, such as marbles can be best removed with a stone retrieval basket, which can be made from a condom. Disk batteries can not be grasped with the FB forceps and snares, but very safely with the powerful magnet attached to the tip of the scope. Overtubes and protective rubber hoods are useful for removing sharp or pointed objects. It is important to test the available grasping accessories on a duplicate of the FB as a "dry run" to determine which accessories will grasp the FB securely.


Functional imaging of the cerebellum and basal ganglia during predictive motor timing in early Parkinson's disease.
The basal ganglia and the cerebellum have both emerged as important structures involved in the processing of temporal information. We examined the roles of the cerebellum and striatum in predictive motor timing during a target interception task in healthy individuals (HC group; n = 21) and in patients with early Parkinson's disease (early stage PD group; n = 20) using functional magnetic resonance imaging. Despite having similar hit ratios, the PD failed more often than the HC to postpone their actions until the right moment and to adapt their behavior from one trial to the next. We found more activation in the right cerebellar lobule VI in HC than in early stage PD during successful trials. Successful trial-by-trial adjustments were associated with higher activity in the right putamen and lobule VI of the cerebellum in HC. We conclude that both the cerebellum and striatum are involved in predictive motor timing tasks. The cerebellar activity is associated exclusively with the postponement of action until the right moment, whereas both the cerebellum and striatum are needed for successful adaptation of motor actions from one trial to the next. We found a general ''hypoactivation'' of basal ganglia and cerebellum in early stage PD relative to HC, indicating that even in early stages of the PD there could be functional perturbations in the motor system beyond striatum.


Effects of high-intensity focused ultrasound on cisplatin-resistant human lung adenocarcinoma in vitro and in vivo.
It is widely accepted that high-intensity focused ultrasound (HIFU) is a minimally invasive treatment option for different tumors, but its roles and the corresponding mechanism in cisplatin (DDP) chemoresistance in lung adenocarcinoma (LA) remain unclear. In this study, we investigated the response of DDP-resistant LA cells to HIFU and its underlying molecular mechanisms using molecular biology techniques. It was found that HIFU exposure inhibited the proliferation of DDP-resistant A549 (A549/DDP) cells through arresting cell cycle at the G1/G0 phase via the Cyclin-dependent pathway and promoting apoptosis in a Bcl-2-dependent manner. Furthermore, the results also showed that HIFU exposure could down-regulate the expressions of MDR1, MRP1, and LRP mRNAs, as well as P-gp, MRP1, and LRP proteins related to drug resistance in A549/DDP cells. In vivo experiments also demonstrated that HIFU could reduce the size and mass of subcutaneously transplanted tumors produced by A549/DDP cells through mediating Cyclin-dependent and Bcl-2-dependent pathways. These results suggested that HIFU treatment could inhibit the proliferation of DDP-resistant lung cancer cells and might be a novel therapeutic method for patients with DDP resistance.


COVID-19 in China: Power, Transparency and Governance in Public Health Crisis.
Public health crises are the "touchstone" to test the ability of national public health crisis governance. The public health crisis in the new era presents new characteristics: systematic, cross-border and uncertainty. The governance dilemma of a public health crisis generally emphasizes the joint participation and communication of different subjects, which is suspected of overlapping and redundancy, and lacks the auxiliary support of major public health crisis events. It dispels the significance of government-level cooperation. The essence of the public health crisis governance system is the chain law of stimulus-response. In combination with COVID-19 development in China, we track down the main reasons for the temporary disruption and the government's response to this major public health crisis. We mainly examine the tension between the centralization of power in China's governance structure and the effectiveness of local governance, and the control of local governments in information disclosure. The response to a public health crisis and the optimization of a decision-making mechanism should build tension between the centralization of power and effectiveness of local governance. It is suggested that government should disclose and share information to the public timely and pay more attention to the core value of order in crisis management.


Diminished LC3-Associated Phagocytosis by Huntington's Disease Striatal Astrocytes.
In recent years the functions of astrocytes have shifted from conventional supportive roles to also include active roles in altering synapses and engulfment of cellular debris. Recent studies have implicated astrocytes in both protective and pathogenic roles impacting Huntington's disease (HD) progression. The goal of this study is to determine if phagocytosis of cellular debris is compromised in HD striatal astrocytes. Primary adult astrocytes were derived from two HD mouse models; the fast-progressing R6/2 and slower progressing Q175. With the use of laser nanosurgery, a single astrocyte was lysed within an astrocyte network. The phagocytic response of astrocytes was observed with phase contrast and by fluorescence microscopy for GFP-LC3 transiently transfected cells. Astrocyte phagocytosis was significantly diminished in primary astrocytes, consistent with the progression of HD in R6/2 and Q175 mouse models. This was defined by the number of astrocytes responding via phagocytosis and by the average number of vesicles formed per cell. GFP-LC3 was found to increasingly localize to phagocytic vesicles over a 20-min imaging period, but not in HD mice, suggesting the involvement of LC3 in astrocyte phagocytosis. We demonstrate a progressive decrease in LC3-associated phagocytosis in HD mouse striatal astrocytes.


[Association between excessive TV viewing time and body mass index among Brazilian adolescents: quantile regression analysis of the National Student Health Survey (PeNSE), 2015].
The study aimed to investigate whether excessive TV viewing time is associated with body mass index (BMI), irrespective of physical activity (PA) and interaction with consumption of ultra-processed foods (UPFs), among Brazilian adolescents. It is a cross-sectional study with data on 13-17-year-old adolescents from the 2015 National Student Health Survey. BMI was the outcome (direct measurement of height and body mass) and exposure was excessive TV viewing time (>2 h/day), daily consumption of UPFs and PA. Quantile regression was used to test associations and interactions. Excessive screen time was associated with higher BMI values (e.g., 25th percentile =0.060 kg/m² versus 95th percentile =0.891 kg/m², p<0.001). After adjusting for PA, the association remained, however, attenuated. The inclusion of UPFs in the model potentiated the association (e.g., 25th percentile =0.062 kg/m² versus 95th percentile =0.956 kg/m², p<0.001). Excessive screen time was associated with BMI percentiles, irrespective of PA - the absence of PA and the presence of UPF consumption affect the upper BMI percentiles more. Actions to reduce excessive TV viewing time, consumption of UPFs and promoting PA are essential to positively impact BMI and its reflexes on the health of Brazilian adolescents.


Enhancement of viability of radiosensitive (PBMC) and resistant (MDA-MB-231) clones in low-dose-rate cobalt-60 radiation therapy.
In the present study, the authors investigated the in vitro behavior of radio-resistant breast adenocarcinoma (MDA-MB-231) cells line and radiosensitive peripheral blood mononuclear cells (PBMC), as a function of different radiation doses, dose rates and postirradiation time kinetics, with a view to the interest of clinical radiotherapy. The cells were irradiated with Co-60, at 2 and 10 Gy and two different exposure rates, 339.56 cGy.min(-1) and the other corresponding to one fourth of the standard dose rates, present over a 10-year period of cobalt therapy. Post-irradiation sampling was performed at pre-established kinetics of 24, 48 and 72 hours. The optical density response in viability assay was evaluated and a morphological analysis was performed. Radiosensitive PBMC showed decrease in viability at 2 Gy, and a more significant decrease at 10 Gy for both dose rates. MDAMB- 231 cells presented viability decrease only at higher dose and dose rate. The results showed MDA-MB-231 clone expansion at low dose rate after 48-72 hours post-radiation. Low dose rate shows a possible potential clinical impact involving decrease in management of radio-resistant and radiosensitive tumor cell lines in cobalt therapy for breast cancer.


The genomic psychiatry cohort: partners in discovery.
The Genomic Psychiatry Cohort (GPC) is a longitudinal resource designed to provide the necessary population-based sample for large-scale genomic studies, studies focusing on Research Domain Criteria (RDoC) and/or other alternate phenotype constructs, clinical and interventional studies, nested case-control studies, long-term disease course studies, and genomic variant-to-phenotype studies. We provide and will continue to encourage access to the GPC as an international resource. DNA and other biological samples and diagnostic data are available through the National Institute of Mental Health (NIMH) Repository. After appropriate review and approval by an advisory board, investigators are able to collaborate in, propose, and co-lead studies involving cohort participants.


Grape polyphenols do not affect vascular function in healthy men.
Data suggest that polyphenol-rich products may improve endothelial function and other cardiovascular health risk factors. Grape and wine contain high amounts of polyphenols, but effects of these polyphenols have hardly been investigated in isolation in randomized controlled studies. Our objective in this study was to test the chronic effect of polyphenol-rich solids derived from either a wine grape mix or grape seed on flow-mediated dilation (FMD). Blood pressure and other vascular function measures, platelet function, and blood lipids were secondary outcomes. Thirty-five healthy males were randomized in a double-blind, placebo-controlled crossover study consisting of three 2-wk intervention periods separated by 1-wk washout periods. The test products, containing 800 mg of polyphenols, were consumed as capsules. At the end of each intervention period, effects were measured after consumption of a low-fat breakfast (~751 kJ, 25% fat) and a high-fat lunch (~3136 kJ, 78% fat). After the low-fat breakfast, the treatments did not significantly affect FMD. The absolute difference after the wine grape solid treatment was -0.4% (95% CI = -1.8 to 0.9; P = 0.77) and after grape seed solids, 0.2% (95% CI = -1.2 to 1.5; P = 0.94) compared with after the placebo treatment. FMD effects after the high-fat lunch and effects on secondary outcomes also showed no consistent differences between both of the grape solids and placebo treatment. In conclusion, consumption of grape polyphenols has no major impact on FMD in healthy men. Future studies should address whether grape polyphenols can improve FMD and other cardiovascular health risk factors in populations with increased cardiovascular risk.


The embodied organization of a real-time fetus: the visible and the invisible in prenatal ultrasound examinations.
Through an analysis of videotaped interactions between healthcare professionals and pregnant women during ultrasound prenatal examinations in Japan, I explore some aspects of sequence organization in which an ultrasound real-time fetus is organized. The ultrasound demonstration of the fetal condition is an intrinsically interactional and distributed achievement. The ultrasound fetus is constructed as a real-time object in a particular technological environment; in this environment, the participants' orientations to spatially separated operational fields, that is, the monitor screen and the woman's abdomen, are exhibited and integrated in the actual course of interaction. In conclusion, the fundamental relation between organizational lived work in a technological environment and the observable features of technology will be suggested.


[Fungal allergy. Part I].
Fungal spores are one of the main groups of allergens. They are present in our environment all year long. The concentration of the spores in July and August is 100-1000 higher than pollens. In contrast with the pollens there is a correlation between the spore count and higher risk of medication use, nocturnal asthma attacks, and hospitalization rates. We have presented the clinical classification of fungi, and characterized main indoor and outdoor allergens. The fungi play also an important role in the sick building syndrome. Fungal allergens are not only aeroallergens, they may also get into our body with food. They may also develop contact reactions. Antibiotic allergy is regarded as one of the classic allergic reactions caused by fungi. The id reactions are caused by the fungi from far infection focuses.


Regulation by S-nitrosylation of the Calvin-Benson cycle fructose-1,6-bisphosphatase in Pisum sativum.
Redox regulation is of great importance in chloroplasts. Many chloroplast enzymes, such as those belonging to the Calvin-Benson cycle (CBC), have conserved regulatory cysteines which form inhibitory disulphide bridges when physiological conditions become unfavourable. Amongst these enzymes, cFBP1, the CBC fructose-1,6-bisphosphatase (FBPase) isoform, is well known to be redox activated by thioredoxin f through the reduction of a disulphide bridge involving Cys153 and Cys173. Moreover, data obtained during recent years point to S-nitrosylation as another redox post-translational modification putatively regulating an increasing number of plant enzymes, including cFBP1. In this study we have shown that the Pisum sativum cFBP1 can be efficiently S-nitrosylated by GSNO and SNAP, triggering the formation of the regulatory disulphide. Using in vivo experiments with P. sativum we have established that cFBP1 S-nitrosylation only occurs during the light period and we have elucidated by activity assays with Cys-to-Ser mutants that this enzyme may be inactivated through the S-nitrosylation of Cys153. Finally, in the light of the new data, we have proposed an extended redox-regulation model by integrating the S-nitrosylation and the TRX f-mediated regulation of cFBP1.


No Role for the Ryanodine Receptor in Regulating Cardiac Contraction?
Cardiac contraction is initiated by Ca(2+) leaving the sarcoplasmic reticulum through the ryanodine receptor (RyR). Although opening of the RyR can be modified by various ligands, these have no maintained effect on contraction. We conclude that modulation of the RyR controls sarcoplasmic reticulum Ca(2+) content rather than cytoplasmic Ca(2+) concentration.


Enhanced secretion of recombinant proteins via signal recognition particle (SRP)-dependent secretion pathway by deletion of rrsE in Escherichia coli.
Although signal recognition particle (SRP)-dependent secretion pathway, which is characterized by co-translational translocation, helps prevent cytoplasmic aggregation of proteins before secretion, its limited capacity for the protein secretion is a major hurdle for utilizing the pathway as an attractive route for secretory production of recombinant proteins. Therefore, we developed an Escherichia coli mutant, whose efficiency of secretion via the SRP pathway was dramatically increased. First, we developed a novel FACS-based screening system by combining a periplasmic display system (PECS) and direct fluorescent labeling with the organoarsenic compound, FlAsH-EDT2 . With this screening system, transposon-insertion library of E. coli was screened, and then we isolated mutants which exhibited higher protein production through the SRP pathway than the parental strain. From the genetic analysis, we found that all isolated mutants had the same mutation-disruption of the 16S rRNA gene (rrsE). The positive effect of rrsE deficiency on protein secretion via the SRP pathway was successfully demonstrated using various model proteins including endogenous SRP-dependent proteins, antibodies, and G protein-coupled receptor. For the large-scale production of IgG and GPCR, we performed fed-batch cultivation with the rrsE-deficient mutant, and very high yields of IgG (0.4 g/L) and GPCR (1.4 g/L) were obtained. Biotechnol. Bioeng. 2016;113: 2453-2461. © 2016 Wiley Periodicals, Inc.


Preoperative thyroid ultrasound is indicated in patients undergoing parathyroidectomy for primary hyperparathyroidism.
Primary hyperaparathyroidism (pHPT) is often accompanied by underlying thyroid pathology that can confound preoperative parathyroid localization studies and complicate intra-operative decision making. The aim of this study was to examine the utility of preoperative thyroid ultrasonography (US) in patients prior to undergoing parathyroidectomy for pHPT. An Institutional Review Board approved prospective study was undertaken from January 2005 through July 2008. All patients with pHPT meeting inclusion criteria (n=94) underwent preoperative thyroid ultrasound in addition to standard (99m)Tc-sestamibi scintigraphy for parathyroid localization. Demographics, operative management and final pathology were examined in all cases. Fifty-four of the 94 patients (57%) were noted to have a thyroid nodule on preoperative US, of which 30 (56%) underwent further examination with fine needle aspiration biopsy. Alteration of the operative plan attributable to underlying thyroid pathology occurred in 16 patients (17%), with patients undergoing either total thyroidectomy (n=9) or thyroid lobectomy (n=7). Thyroid cancer was noted in 33% of patients undergoing thyroid resection, and 6% of all patients with HPT. The routine utilization of preoperative thyroid ultrasound in patients prior to undergoing parathyroid surgery for pHPT is indicated. The added information from this non-invasive modality facilitates timely management of co-incidental, and sometimes malignant, thyroid pathology.


Effects of sunscreens and a DNA excision repair enzyme on ultraviolet radiation-induced inflammation, immune suppression, and cyclobutane pyrimidine dimer formation in mice.
Exposure of skin to ultraviolet (UV) radiation inhibits the induction of delayed-type hypersensitivity (DTH) responses initiated at a distant, unirradiated site. Recent studies attributed this form of immune suppression to DNA damage in the form of cyclobutane pyrimidine dimers (CPD). In the present study, we investigated the protective defects of sunscreens on UV-induced systemic suppression of DTH to Candida albicans, inflammation, and DNA damage. The photoprotective effects of sunscreen preparations containing 8% octyl-N-dimethyl-p-aminobenzoate, 7.5% 2-ethylhexyl-p-methoxycinnamate, or 6% benzophenone-3 were studied in C3H mice exposed to a single dose of 500 mJ/cm2 UVB radiation from FS40 sunlamps. Inflammation was determined by the amount of skin edema at the site of UV irradiation, and DNA damage was assessed by measuring the frequency of endonuclease-sensitive sites in the epidermis. Application of the sunscreens before UV irradiation gave 75-97% protection against UV-induced edema, 67-91% protection against formation of CPD, but only 30-54% protection against suppression of DTH. In contrast, the topical application of liposomes containing a CPD-specific DNA repair enzyme immediately after UV irradiation resulted in 82% protection against suppression of DTH, but at best, 39% protection against skin edema. These findings demonstrate that sunscreens give less protection against UV-induced immune suppression than against skin edema and CPD formation. Furthermore, they suggest that less DNA damage is required to cause UV-induced immune suppression than to cause sunburn.


Changing emotional visual and auditory memories: are modality-matched dual-tasks more effective?
Clinical and laboratory studies have demonstrated that executing a demanding dual-task while recollecting emotional memories weakens the emotional intensity and vividness of these memories. While this approach is generally effective, there is room for improvement. According to multi-component working memory theories, the effectiveness of dual-tasks may be improved by loading specifically the same sensory modality of the emotional memories. So far, however, the evidence for this idea is mixed. In the current report, this idea was tested in a pilot study (N = 36) and a pre-registered experiment (N = 60) by exposing participants to pictures of the International Affective Picture System database and to sounds of the International Affective Digital Sounds database, thus creating single-modality emotional memories. Using a within-subjects design, participants had to recollect their memories of the sounds and pictures while executing a visually-demanding task (i.e. identifying visual letters), an auditory-demanding task (i.e. identifying auditory letters), or no task. Across both studies, we only found limited evidence for modality-specific effects of dual-tasks on single-modality emotional memories. We discuss the relevance of our findings for working memory theories of memory change and therapeutic practices.


Similarities and Differences among Protein Dynamics Studied by Variable Temperature Nuclear Magnetic Resonance Relaxation.
Understanding and describing the dynamics of proteins is one of the major challenges in biology. Here, we use multifield variable-temperature NMR longitudinal relaxation (R1) measurements to determine the hierarchical activation energies of motions of four different proteins: two small globular proteins (GB1 and the SH3 domain of α-spectrin), an intrinsically disordered protein (the C-terminus of the nucleoprotein of the Sendai virus, Sendai Ntail), and an outer membrane protein (OmpG). The activation energies map the motions occurring in the side chains, in the backbone, and in the hydration shells of the proteins. We were able to identify similarities and differences in the average motions of the proteins. We find that the NMR relaxation properties of the four proteins do share similar features. The data characterizing average backbone motions are found to be very similar, the same for methyl group rotations, and similar activation energies are measured. The main observed difference occurs for the intrinsically disordered Sendai Ntail, where we observe much lower energy of activation for motions of protons associated with the protein-solvent interface as compared to the others. We also observe variability between the proteins regarding side chain 15N relaxation of lysine residues, with a higher activation energy observed in OmpG. This hints at strong interactions with negatively charged lipids in the bilayer and provides a possible mechanistic clue for the "positive-inside" rule for helical membrane proteins. Overall, these observations refine the understanding of the similarities and differences between hierarchical dynamics in proteins.


The GrlR-GrlA regulatory system coordinately controls the expression of flagellar and LEE-encoded type III protein secretion systems in enterohemorrhagic Escherichia coli.
The gene function of the locus of enterocyte effacement (LEE) is essential for full virulence of enterohemorrhagic Escherichia coli (EHEC). Strict control of LEE gene expression is mediated by the coordinated activities of several regulatory elements. We previously reported that the ClpX/ClpP protease positively controls LEE expression by down-regulating intracellular levels of GrlR, a negative regulator of LEE gene expression. We further revealed that the negative effect of GrlR on LEE expression was mediated through GrlA, a positive regulator of LEE expression. In this study, we found that the FliC protein, a major component of flagellar filament, was overproduced in clpXP mutant EHEC, as previously reported for Salmonella. We further found that FliC expression was reduced in a clpXP grlR double mutant. To determine the mediators of this phenotype, FliC protein levels in wild-type, grlR, grlA, and grlR grlA strains were compared. Steady-state levels of FliC protein were reduced only in the grlR mutant, suggesting that positive regulation of FliC expression by GrlR is mediated by GrlA. Correspondingly, cell motility was also reduced in the grlR mutant, but not in the grlA or grlR grlA mutant. Because overexpression of grlA from a multicopy plasmid strongly represses the FliC level, as well as cell motility, we conclude that GrlA acts as a negative regulator of flagellar-gene expression. The fact that an EHEC strain constitutively expressing FlhD/FlhC cannot adhere to HeLa cells leads us to hypothesize that GrlA-dependent repression of the flagellar regulon is important for efficient cell adhesion of EHEC to host cells.


[Discrepant outcome of intrauterine listeria infection in dichorionic twins].
BACKGROUND AND CASE REPORT: We report on a case of fetal Listeriosis in a dichorionic twin pregnancy where both placentae, but only one of the twins were infected. While the firstborn child showed no infection and remained healthy until today, the other newborn had all clinical signs of granulomatosis infantiseptica and died despite of immediate resuscitation immediately after delivery. This discrepant course with Listeriosis in twins underlines, that fetal factors influence the clinical outcome in placental Listeriosis. The reasons for the infection of only one twin and the avoidance of the other twin remain unclarified. We speculate that immunologic mechanisms or the presence of meconium-stained amniotic fluid may play an important role for intrauterine infection with Listeriosis.


Beyond the label: current evidence and future directions for the interrelationship between electronic cigarettes and mental health.
Electronic cigarette use has dramatically increased over the last decade. With this recent technological development and wide range of constituents in various products, putative adverse effects on the brain and body have been largely unexplored. Here, we review current evidence linking electronic nicotine cigarette use with potential health consequences and provide evidence supporting an association between drug use and depression in humans. We also examine the biological effects of individual constituents in electronic cigarette aerosols, which include labeled ingredients, such as propylene glycol, vegetable glycerin, nicotine, and flavorants, as well as unlabeled ingredients found in the aerosols, such as carbonyls and heavy metals. Lastly, we examine the effects of electronic cigarette use on endogenous metabolism via changes in cytochrome P450 enzymes, which can thereby impact therapeutic outcomes. While the current evidence offers insight into the potential effects of electronic cigarette use on biological processes, further studies are necessary to determine the long-term clinical relevance of aerosol inhalation.


Contemporary management of dyslipidemia in high-risk patients: targets still not met.
Our objective was to evaluate treatment patterns and the attainment of current National Cholesterol Education Program (NCEP)-recommended lipid targets in unselected high-risk ambulatory patients. Between December 2001 and December 2004, the prospective Vascular Protection and Guidelines Oriented Approach to Lipid Lowering Registries recruited 8056 outpatients with diabetes, established cardiovascular disease (CVD), or both, who had a complete lipid profile measured within 6 months before enrollment. The primary outcome measure was treatment success, defined as the achievement of LDL-cholesterol<2.6 mmol/L (100 mg/dL) according to NCEP guidelines. We examined patient characteristics and use of lipid-modifying therapy in relation to treatment outcome, which included the recently proposed optional LDL-cholesterol target (<1.8 mmol/L [70 mg/dL]) for very high-risk patients. Overall, 78.2% of patients were treated with a statin and 51.2% had achieved the recommended LDL-cholesterol target. Treatment success rate was highest in diabetic patients with CVD (59.6%), followed by nondiabetic patients with CVD (51.8%), and lowest (44.8%) in diabetic patients without CVD (P<.0001). Compared with untreated patients, those on statins were more likely to achieve target (34.4% vs 55.9%, P<.0001). Of the patients who failed to meet target, only 9.9% were taking high-dose statin, while 29.3% were not prescribed any statin therapy. Among very high-risk patients, 20.8% attained the optional LDL-cholesterol goal. In multivariable analysis, advanced age, male sex, diabetes, coronary artery disease, coronary revascularization, and use of statin were associated with treatment success (all P<.0001). Despite the well-established benefits of available lipid-modifying drugs, current management of dyslipidemia continues to be suboptimal, with a substantial proportion of patients failing to achieve guideline-recommended lipid targets. There remains an important opportunity to improve the quality of care for these high-risk patients.


Identification of Candida glabrata by a 30-second trehalase test.
Rapid (30-s) trehalase tests done with material from colonies of 482 yeasts suspended in a drop of trehalose solution on a commercially supplied glucose test strip were positive for 225 (99.1%) of 227 Candida glabrata isolates grown on either of two differential media, Candida ID medium or CandiSelect medium. The test was positive for only 3 (1.2%) and 12 (4.7%) of 255 isolates of other medically important yeast species grown on the same two media, respectively. A rapid maltase test done with a subset of 255 yeast isolates was negative for all but 1 of 64 trehalase-positive C. glabrata isolates, raising the specificity of the rapid testing for C. glabrata to 98.4 to 100%, depending on the isolation medium used. Rapid trehalase and maltase tests done independently in two laboratories with 217 yeast isolates showed sensitivities of 96.0 to 98.0% and specificities of 98.2 to 99.4% for identification of C. glabrata from colonies grown on Candida ID medium. The specificity was much lower because of frequent false-positive trehalose test results when the source of colonies was Sabouraud agar formulated with 4% glucose. We conclude that direct recognition of C. albicans as blue colonies on Candida ID isolation medium coupled with the performance of the 30-s trehalase and maltase tests for C. glabrata among the white colonies on this medium will allow the rapid presumptive identification of the two yeast species most commonly encountered in clinical samples.


Effect of submucosal injection of dexamethasone on postoperative discomfort after third molar surgery: a prospective study.
To evaluate the relative ability of 4 mg dose of intraoperative dexamethasone, administered submucosally, to reduce the postoperative discomfort after third molar surgery. A total of 100 patients requiring surgical removal of a single mandibular third molar were included. The experimental group (50) received dexamethasone 4 mg as submucosal injection and control group (50) received no drugs. The maximum interincisal distance and facial contours were measured at baseline and at postsurgery days 2 and 7. The measurement of pain was done using visual analog scale (VAS). None of the patients developed wound infection or any serious postoperative complications. Postoperative edema tended to be less severe on the second postoperative day in the experimental group and the result was statistically significant. There were no significant differences in the reduction of pain and trismus between the two groups studied. Submucosal administration of dexamethasone sodium phosphate (4 mg) results in reduction of postoperative edema, comparable with or greater than other routes of administration. Presumably, a higher effective drug concentration at the site of injury without loss due to distribution to other compartments may be achieved, and the submucosal route is convenient for both the surgeon and the patient. Submucosal route of administration of dexamethasone (4 mg) is effective in reducing postoperative discomfort after third molar surgeries.


Stent-Assisted Coil Embolization of Vertebrobasilar Dissecting Aneurysms: Procedural Outcomes and Factors for Recanalization.
Outcomes of stent-assisted coil embolization (SACE) have not been well established in the setting of vertebrobasilar dissecting aneurysms (VBDAs) due to the low percentage of cases that need treatment and the array of available therapeutic options. Herein, we presented clinical and radiographic results of SACE in patients with VBDAs. A total of 47 patients (M:F, 30:17; mean age ± SD, 53.7 ± 12.6 years), with a VBDA who underwent SACE between 2008 and 2014 at two institutions were evaluated retrospectively. Medical records and radiologic data were analyzed to assess the outcome of SACE procedures. Cox proportional hazards regression analysis was conducted to determine the factors that were associated with aneurysmal recanalization after SACE. Stent-assisted coil embolization technically succeeded in all patients. Three cerebellar infarctions occurred on postembolization day 1, week 2, and month 2, but no other procedure-related complications developed. Immediately following SACE, 25 aneurysms (53.2%) showed no contrast filling into the aneurysmal sac. During a mean follow-up of 20.2 months, 37 lesions (78.7%) appeared completely occluded, whereas 10 lesions showed recanalization, 5 of which required additional embolization. Overall recanalization rate was 12.64% per lesion-year, and mean postoperative time to recanalization was 18 months (range, 3-36 months). In multivariable analysis, major branch involvement (hazard ratio [HR]: 7.28; p = 0.013) and the presence of residual sac filling (HR: 8.49, p = 0.044) were identified as statistically significant independent predictors of recanalization. No bleeding was encountered in follow-up monitoring. Stent-assisted coil embolization appears feasible and safe for treatment of VBDAs. Long-term results were acceptable in a majority of patients studied, despite a relatively high rate of incomplete occlusion immediately after SACE. Major branch involvement and coiled aneurysms with residual sac filling may predispose to recanalization.


Reattachment and orthodontic extrusion in the management of an incisor crown-root fracture: a case report.
This article describes the management of a case of oblique crown root fracture of a central incisor where the fractured fragment was used as a temporary replacement crown in order to maintain esthetics during the period of orthodontic extrusion of the remaining root. Placement of the final restoration after orthodontic extrusion resulted in good esthetics and periodontal health postoperatively. The merits and demerits of this treatment modality are discussed.


Successful endovascular treatment of a spinal dural arteriovenous fistula with trisacryl gelatin microspheres.
We report on the endovascular treatment of a spinal dural arteriovenous fistula (SDAVF) in a 50-year-old man, who presented with rapidly progressive paraparesis of lower extremities. Standard treatment of SDAVFs is by either embolization with liquid adhesive agents like NBCA or microsurgery. In our case the fistula was successfully occluded by application of trisacryl gelatin microspheres (TGM). Due to a small size of the feeding intercostal artery a superselective catheterization of the fistula itself by a microcatheter could not be performed. The particles were hence delivered through a diagnostic spinal catheter positioned within the proximal part of the feeding intercostal artery. Control MRI five weeks after intervention showed normalized perimedullary veins, which were dilated and tortuous initially. It also displayed a clinically silent focal ischaemia within the spongiosa of an adjacent hemi-vertebra due to particle passage through intersegmental vascular collaterals. A control angiogram confirmed the disappearance of the fistula. The patient underwent physiotherapy and experienced significant improvement of his gait within three months after intervention.


[Child's status in modern-day Finnish society].
Despite the fact that the child's status is protected in the Finnish legislation, the development is taking a turn for the worse. Health inequalities among children are a matter of concern, and an increasing number of children need child protection. The best interest of the child is not taken into account in decision-making or everyday life in a manner consistent with children's rights. In view of the dependency ratio and public-sector finances of the future years, it is essential that the development of children and the young will be cared for in a manner allowing them to grow into healthy adults.


Breast cancer clusters in the northeast United States: a geographic analysis.
High breast cancer mortality rates have been reported in the northeastern part of the United States, with recent attention focused on Long Island, New York. In this study, the authors investigate whether the high breast cancer mortality is evenly spread over the Northeast, in the sense that any observed clusters of deaths can be explained by chance alone, or whether there are clusters of statistical significance. Demographic data and age-specific breast cancer mortality rates for women were obtained for all 244 counties in 11 northeastern states and for the District of Columbia for 1988-1992. A recently developed spatial scan statistic is used, which searches for clusters of cases without specifying their size or location ahead of time, and which tests for their statistical significance while adjusting for the multiple testing inherent in such a procedure. The basic analysis is adjusted for age, with further analyses examining how the results are affected by incorporating race, urbanicity, and parity as confounding variables. There is a statistically significant and geographically broad cluster of breast cancer deaths in the New York City-Philadelphia, Pennsylvania, metropolitan area (p = 0.0001), which has a 7.4% higher mortality rate than the rest of the Northeast. The cluster remains significant when race, urbanicity, and/or parity are included as confounding variables. Four smaller subclusters within this area are also significant on their own strength: Philadelphia with suburbs (p = 0.0001), Long Island (p = 0.0001), central New Jersey (p = 0.0001), and northeastern New Jersey (p = 0.0001). The elevated breast cancer mortality on Long Island might be viewed less as a unique local phenomenon and more as part of a more general situation involving large parts of the New York City-Philadelphia metropolitan area. The several known and hypothesized risk factors for which we could not adjust and that may explain the detected cluster are most notably age at first birth, age at menarche, age at menopause, breastfeeding, genetic mutations, and environmental factors.


Central and peripheral circadian clocks and their role in Alzheimer's disease.
Molecular and cellular oscillations constitute an internal clock that tracks the time of day and permits organisms to optimize their behaviour and metabolism to suit the daily demands they face. The workings of this internal clock become impaired with age. In this review, we discuss whether such age-related impairments in the circadian clock interact with age-related neurodegenerative disorders, such as Alzheimer's disease. Findings from mouse and fly models of Alzheimer's disease have accelerated our understanding of the interaction between neurodegeneration and circadian biology. These models show that neurodegeneration likely impairs circadian rhythms either by damaging the central clock or by blocking its communication with other brain areas and with peripheral tissues. The consequent sleep and metabolic deficits could enhance the susceptibility of the brain to further degenerative processes. Thus, circadian dysfunction might be both a cause and an effect of neurodegeneration. We also discuss the primary role of light in the entrainment of the central clock and describe important, alternative time signals, such as food, that play a role in entraining central and peripheral circadian clocks. Finally, we propose how these recent insights could inform efforts to develop novel therapeutic approaches to re-entrain arrhythmic individuals with neurodegenerative disease.


[Peduncular hallucination in brain stem cavernous angioma: a case report].
A case of brain stem cavernous angioma associated with peduncular hallucination is reported. A 68-year-old man was admitted with complaints of left sensory disturbance. On the evening of the first hospital day, he developed vividly formed hallucinations. The hallucinations were accompanied by sleep disturbance and agitation. At approximately 2:00 am, he began to feel as though he could see his daughter watching television in a room. At one point he saw vivid pictures. MRI revealed a round mass with a mixed signal intensity core surrounded by a low signal intensity rim in the right brain stem. On the 10th hospital day, there were no hallucinations. On October 12th suboccipital craniectomy was performed and cavernous angioma and the surrounding hematoma was partially removed. Soon after that the patient was discharged with improvement of both ocular movement and left-sided sensory disturbance.


[Four cases of persistent hyperplastic primary vitreous].
We evaluated four cases of persistent hyperplastic primary vitreous (PHPV) encountered at Nagoya City University Hospital in 1995. PHPV was seen unilaterally in three cases and bilaterally in one. The series comprised two males and two females, ranging in age from three to eight months, with an average of 4.8 months. Case 1 had a white strand running from the optic disc to the posterior surface of the lens in the left eye. Case 2 showed leukocoria in the right eye and central corneal opacity in the left. Magnetic resonance imaging (MRI) revealed total retinal detachment in both eyes. Case 3 exhibited retinal folds running from the optic disc to the posterior surface of the lens in the left eye. Case 4 showed elongation of the ciliary processes and leukocoria in the right eye. Ipsilateral total retinal detachment was seen in MRI. Three eyes of two cases were microphthalmic. Associated ocular anomalies included, posterior embryotoxon, sclerocornea, hypoplasia of the iris stroma and peripapillary staphyloma. There were accompanying systemic anomalies such as arachnoidal cyst, syndactyly, microcephalus, heart anomalies, pulmonary atresia and asplenia. Patients with PHPV should be carefully examined for the possible presence of other ocular and systemic anomalies caused by neural crest disorders.


Three-dimensional structures of pathogenic and saprophytic Leptospira species revealed by cryo-electron tomography.
Leptospira interrogans is the primary causative agent of the most widespread zoonotic disease, leptospirosis. An in-depth structural characterization of L. interrogans is needed to understand its biology and pathogenesis. In this study, cryo-electron tomography (cryo-ET) was used to compare pathogenic and saprophytic species and examine the unique morphological features of this group of bacteria. Specifically, our study revealed a structural difference between the cell envelopes of L. interrogans and Leptospira biflexa involving variations in the lipopolysaccharide (LPS) layer. Through cryo-ET and subvolume averaging, we determined the first three-dimensional (3-D) structure of the flagellar motor of leptospira, with novel features in the flagellar C ring, export apparatus, and stator. Together with direct visualization of chemoreceptor arrays, DNA packing, periplasmic filaments, spherical cytoplasmic bodies, and a unique "cap" at the cell end, this report provides structural insights into these fascinating Leptospira species.


An appraisal of nuclear organisation in interphase embryonic fibroblasts of chicken, turkey and duck.
Determining the nuclear 'addresses' of chromosome territories is a well-documented means of assaying for nuclear organisation in many cell types and species. Data in avian species are however limited at best, despite the pivotal role played by birds (particularly chickens) in agriculture, and as model organisms in developmental biology. That is, studies have hitherto focussed mostly on mammals (especially humans) and have demonstrated the importance of chromosome territory positioning in embryology, disease and evolution. Thus a detailed study of nuclear organisation in many species, many cell types and many developmental stages in birds is warranted, however, before this is achieved, 'baseline' needs to be established to determine precisely the relative locations of chromosome territories in at least 1 cell type of at least 1 bird. With this in mind we hybridised FISH probes from chicken chromosomes 1-28 to embryonic fibroblast nuclei, determining nuclear addresses using a newly developed plug-in to the image analysis package ImageJ. In our experience, evenly spaced representative BAC clones yielded more consistent results than hybridisation of chromosome paints. Results suggested that chromosome territory distribution best fitted a chromosome size-based (rather than gene density-based) pattern. Identical BAC clones were then hybridised to turkey and duck in a comparative genomic strategy. Observations were consistent with those seen in chicken (although, less well-defined in duck), providing preliminary evidence of conservation throughout evolution.


Plasmonics metalens independent from the incident polarizations.
An ultra-thin, planar, broadband metalens composed of L-shaped gap antennas on a thin gold film has been designed, which is suitable for both circular and X/Y linear polarizations focusing simultaneously. The phase discontinuity of the cross-polarized transmisson light can be manipulated by the length and width of the L-shaped gap antenna accurately. The designed planar metalens posses a strong focusing ability over a large wavelength range, and the size of the focus spot is in sub-wavelength scale. The focal lengths change from 13 to 7 um with incident wavelength from 750 to 1300 nm, and the cause of dispersion is explained and analyzed in detail. The designed metalens can work very well at a wide incident angles of 0~45°. Most importantly, its unique focusing ability that is independent of the incident polarizations will greatly promotes the practical applications and developments of the metasurfaces.


[Classification of gastroesophageal reflux disease and gastritis].
The gastroesophageal reflux disease (GORD) is frequent and causes by retrograde flow of the gastric content through incompetent gastroesophageal junction. Epidemiological studies have proved that GORD is associated with hearburn in high prevalence. In western countries several studies reported that 20-40% of adult population experience heartburn symptoms at least once in the year, approximately 10% have symptoms weekly and 5% daily. Esophagitis was objectively defined as a mucosal damage and it was endoscopically verificated in 25% of patients. Indeed, GORD symptoms and esophagitis are in poor correlation and less than half of patients with heartburn symptoms had esophagitis on endoscopy. From 1989, Savary Monniér and Metaplasia-Ulcer-Stricture-Erosion (MUSE) endoscopically classification is in use. From 1994, LA (Los Angeles) classification of reflux disease is also in use by endoscopists. During its life cycle, gastric mucosa is exposed to different harmful agents and its response is restitution "ad integrum" on the beginning and at the end of process. First line defence is mucuse barrier which prevent contact between epithelial cell and possible irritant. Important role in mucuse layer plays prostaglandins. After several classification systems previously used, in 1991 Price introduced Sydney system gradation and gastritis classification. Pointing out importance of topographical differences in gastritis distribution, system has introduced 5 histological variations in its Morphological section: chronic inflammation, neutrophylic activity, glandular atrophy, intestinal metaplasy and H. pylori colonisation, with 4 points grading.


Collection and transfusion of blood and blood components in the United States, 1989.
To probe recent trends in transfusion practice and their effect on the adequacy of the national blood resource, transfusions and collections in the United States in 1989 were studied, by using data shared by the American Association of Blood Banks, the American Red Cross, and the Council of Community Blood Centers, together with results from a sample survey of the 3600 hospitals that were not members of the national organizations. Statistical methods were used to estimate national activities. The total US supply of blood in 1989 was 14,229,000 units, an increase of 1.2 percent over the supply in 1987. Red cell transfusions were 12,059,000 units. A total of 3,159,000 patients underwent transfusion with whole blood and/or red cells (mean, 3.8 units/patient). Preoperative autologous deposits of 655,000 units by 310,000 patients represented an increase of 65 percent over the level in 1987. However, only 356,000 units (54%) were transfused to the patients who preoperatively deposited them; of the remainder, 13,000 units were crossed over for transfusion to other patients, while 286,000 units were never used. Directed donations, 350,000 units, were provided for 130,000 intended recipients, but only 97,000 units (28%) were transfused to their intended recipients; of the balance, 59,000 units (17%) were crossed over and 194,000 units (55%) were never transfused. Total platelet transfusions were equivalent to 7,258,000 units in 1989, for an increase of 13.7 percent over totals in 1987.


Inhibitory Effect of Genistein on PLC/PRF5 Hepatocellular Carcinoma Cell Line.
Natural compounds including flavonoids like genistein (GE) are able to inhibit cell proliferation and induce apoptosis. GE is the main representative of these groups. GE inhibits carcinogenic tumors such as colon, stomach, lung, and pancreas tumors. The aim of the present study was to analyze the apoptotic effect of GE in the hepatocellular carcinoma (HCC) PLC/PRF5 cell line. Cells were treated with various doses of GE (1, 5, 10, 25, 50, 75, and 100 μM/L) at different times (24, 48, and 72 h) and the MTT assay was commonly used. Furthermore, cells were treated with single dose of GE (25 μM) at different times and flow cytometry was performed. GE inhibited the growth of liver cancer cells significantly with a time- and dose-dependent manner. The percentage of living cells in GE treatment groups with a concentration of 25 μM at different times were 53, 48 and 47%, respectively (P < 0.001). Result of flow cytometry demonstrated that GE at a 25 μM concentration induces apoptosis significantly in a time-dependent manner. The percentage of apoptotic cells at different times were 44, 56, and 60%, respectively (P < 0.001). GE can significantly inhibit the growth of HCC cells and plays a significant role in apoptosis of this cell line.


Evaluation of trovafloxacin in the treatment of Klebsiella pneumoniae lung infection in tumour-bearing mice.
Trovafloxacin, a new trifluoroquinolone, was evaluated for its therapeutic efficacy against Klebsiella pneumoniae lung infection in tumour (P388 murine leukaemia cells)-bearing mice, treated with or without a chemotherapeutic agent, daunorubicin (DNR) and in mice without tumour. Its activity was compared with ciprofloxacin and cephazolin. The effect on therapeutic efficacy of the addition of recombinant granulocyte colony stimulating factor (rGCSF) was also examined. Our study showed that both quinolones successfully cured pneumonia owing to infection with K. pneumoniae in mice without tumours but that all antibiotics failed in tumour-bearing mice if DNR was withheld. Substantial differences were noted in DNR-treated tumour-bearing mice with infection-the cure rate with trovafloxacin was 91% whereas the cure rate with ciprofloxacin or cephazolin was 57%. Addition of rGCSF to ciprofloxacin did not substantially improve its efficacy (when assessed by protection against death owing to infection; the survival rate was 41%). Trovafloxacin cure rates ranged from 80 to 90% whether or not rGCSF was added to the treatment regimen. Our results suggest that prior cancer chemotherapy had no adverse effect on the therapeutic efficacy of trovafloxacin, and that trovafloxacin may be a promising therapeutic agent for treatment of bacterial infections in the presence of leucopenia.


Technology for community health alliances.
A community health alliance brings together divergent interests within a community for the betterment of personal and population health. In this report we describe how a community responsive strategy in Chicago is facilitating the improvement of healthcare by providing local information of what needs to be done, supporting change at the practice level to meet these needs, and initiating community-wide approaches to manage prevalent and important needs without waiting for direct involvement of health professionals.


Desmoid tumor of the hand: a case report.
Extra-abdominal desmoid tumors are extremely rare in the hand. These tumors do not metastasize; however, they are potentially locally invasive and have extremely high local recurrence rates after surgical excision with reports of up to 78% recurrence, specifically in the hand. We describe the first case of a desmoid tumor originating from the extensor mechanism of a digit and discuss our treatment approach. In addition, a literature review performed shows a male predominance of desmoid tumors in the hand in the 30- to 50-year-old age group. Current evidence supports aggressive early wide surgical excision and reconstruction to preserve function.


Busulfan use in hematopoietic stem cell transplantation: pharmacology, dose adjustment, safety and efficacy in adults and children.
Busulphan (1, 4-bis [methanesulfonyl-y] butane) is a bi-functional alkylating agent that, in combination with cyclophosphamide, has been commonly used in conditioning regimens before hematological stem cell transplantation (HSCT) for nearly 20 years. Busulfan has a very narrow therapeutic index, and acute toxicity may be related to absorption and disposition of the drug and metabolites. Precise delivery of the oral formulation is compromised by erratic gastrointestinal absorption, particularly in infants and small children. An intravenous busulfan formula was approved nearly 40 years after the approval of the oral formulation. Busulfan levels expressed as the area under the concentration-time curve (AUC) higher than 1500 microM* minute were reported to increase the risk of developing veno-occlusive disease (VOD), while low levels may result in engraftment failure or disease relapse. VOD occurs in 11-40% of patients undergoing HSCT and is associated with death in 3.3% of patients. Measurement of individual plasma busulfan levels during oral or intravenous dosing to obtain an AUC is likely to provide the necessary elements to monitor the drug disposition, ensuring efficacy and preventing toxicity of patients undergoing HSCT. It is also important to consider the busulfan drug-drug interactions and adverse drug reactions that can develop during the therapeutic process. Busulfan therapeutic drug monitoring and dose-adjustment should be performed in specialized laboratories staffed by well-trained personnel.


Niobium-complex-based syntheses of sodium niobate nanowires possessing superior photocatalytic properties.
Sodium niobates with nanowire morphology (NaNbO(3)-NW) were synthesized in a large scale by use of a niobium oxooxalate complex as the starting material. This NaNbO(3)-NW showed definitely enhanced photocatalytic activity for H(2) or O(2) evolution in the presence of sacrificial reagents and an overall water splitting under UV-light irradiation, as compared with a bulky counterpart (NaNbO(3)-B). This is the first example that an overall water splitting into H(2) and O(2) proceeded on the semiconductor nanowire photocatalyst.


Epigenetic silencing of the oncogenic miR-17-92 cluster during PU.1-directed macrophage differentiation.
The oncogenic cluster miR-17-92 encodes seven related microRNAs that regulate cell proliferation, apoptosis and development. Expression of miR-17-92 cluster is decreased upon cell differentiation. Here, we report a novel mechanism of the regulation of miR-17-92 cluster. Using transgenic PU.1(-/-) myeloid progenitors we show that upon macrophage differentiation, the transcription factor PU.1 induces the secondary determinant Egr2 which, in turn, directly represses miR-17-92 expression by recruiting histone demethylase Jarid1b leading to histone H3 lysine K4 demethylation within the CpG island at the miR-17-92 promoter. Conversely, Egr2 itself is targeted by miR-17-92, indicating existence of mutual regulatory relationship between miR-17-92 and Egr2. Furthermore, restoring EGR2 levels in primary acute myeloid leukaemia blasts expressing elevated levels of miR-17-92 and low levels of PU.1 and EGR2 leads to downregulation of miR-17-92 and restored expression of its targets p21CIP1 and BIM. We propose that upon macrophage differentiation PU.1 represses the miR-17-92 cluster promoter by an Egr-2/Jarid1b-mediated H3K4 demethylation mechanism whose deregulation may contribute to leukaemic states.


Endocrine mechanisms of primate life history trade-offs: growth and reproductive maturation in vervet monkeys.
Life history theory predicts that the timing of maturation will result from a trade-off between growth and the age of first reproduction. This trade-off and its mechanisms of action are still poorly understood in many species and have not been well studied at the individual level. This study examined hypothesized trade-offs between growth and reproductive maturation in wild populations of vervet monkeys (Cercopithecus aethiops) from Kenya, East Africa. Individuals were sampled from four populations in widely separated sites differing in temperature, altitude, and rainfall. Biological samples and morphometric measures were collected from 50 adult males, 83 adult females, and 225 juveniles. Gonadal steroids and leptin levels were analyzed by radioimmunoassay of sera from 136 juvenile males and 90 juvenile females. Cross-sectional profiles of morphometric and endocrine data were used to assess the onset and cessation of growth in relation to sexual maturation. Gonadal steroids were used to assess sexual maturation and breeding onset. Leptin was used as an index of nutritional state. Estimates of mortality were derived from population age-structure. Across populations, higher resource productivity and nutrient status were associated with more rapid growth. Shorter growth duration was associated with earlier reproductive onset. These findings provide support for models of trade-offs between the timing of growth completion and reproductive onset, but they are contradicted by the evidence that reproduction precedes the cessation of growth in these populations. The biphasic actions of estradiol provide an alternative model and mechanism for the growth-reproduction trade-off.


Mcl-1 downregulation by pro-inflammatory cytokines and palmitate is an early event contributing to β-cell apoptosis.
Pancreatic β-cell apoptosis is a key feature of diabetes mellitus and the mitochondrial pathway of apoptosis is a major mediator of β-cell death. We presently evaluated the role of the myeloid cell leukemia sequence 1 (Mcl-1), an antiapoptotic protein of the Bcl-2 family, in β-cells following exposure to well-defined β-cell death effectors, for example, pro-inflammatory cytokines, palmitate and chemical endoplasmic reticulum (ER) stressors. All cytotoxic stresses rapidly and preferentially decreased Mcl-1 protein expression as compared with the late effect observed on the other antiapoptotic proteins, Bcl-2 and Bcl-xL. This was due to ER stress-mediated inhibition of translation through eIF2α phosphorylation for palmitate and ER stressors and through the combined action of translation inhibition and JNK activation for cytokines. Knocking down Mcl-1 using small interference RNAs increased apoptosis and caspase-3 cleavage induced by cytokines, palmitate or thapsigargin, whereas Mcl-1 overexpression partly prevented Bax translocation to the mitochondria, cytochrome c release, caspase-3 cleavage and apoptosis induced by the β-cell death effectors. Altogether, our data suggest that Mcl-1 downregulation is a crucial event leading to β-cell apoptosis and provide new insights into the mechanisms linking ER stress and the mitochondrial intrinsic pathway of apoptosis. Mcl-1 is therefore an attractive target for the design of new strategies in the treatment of diabetes.


Surgical treatment of locally recurrent rectal cancer: a narrative review.
To summarize the recent literature on surgical treatment of locally recurrent rectal cancer (LRRC). LRRC is a heterogeneous disease that requires a multidisciplinary treatment approach. The treatment and prognosis depend on the site and type of recurrence. Radical resection remains the primary method for achieving long-term survival and improving symptom control. Preoperative chemoradiotherapy can reduce tumor volume and improve the R0 resection rate. Surgeons must clearly understand pelvic anatomy, develop a detailed preoperative plan, adopt a multidisciplinary approach for the surgical resection of the tumor as well as any invaded soft tissues, vessels, and bones, and ensure proper reconstruction. However, extended radical surgery often leads to a higher risk of postoperative complications and a low quality of life. We searched English-language articles with keywords "locally recurrent rectal cancer", "surgery" and "multidisciplinary team" in PubMed published between January 2000 to October 2020. LRRC is a complex problem. Long-term survival is not impossible following multidisciplinary treatment in appropriately selected LRRC patients. The management of LRRC relies on a specialist team that determines the biological behavior of the tumor and evaluates treatment options through multidisciplinary discussions, thereby balancing the surgical costs and benefits, alleviating postoperative complications, and improving patients' quality of life.


Encoding signs of orofacial neuropathic pain from facial expressions in mice.
To evaluate the effects of mental nerve injury in the facial reactions elicited by mechanical stimulation of different intensities and detect and quantify spontaneous facial pain-like expressions during a period free of stimuli, as signs of evoked and spontaneous pain in a mouse model for neuropathic orofacial pain. We recorded mouse heads in a fixed position during a stimulus-free period and with mechanical stimulation with 3 different Von Frey filaments. We extracted the Histograms of Oriented Gradients of each frame of the video recordings to be compared with a prototypical pain-like facial expression. The similarity score was then used to register and quantify the percentage of spontaneous pain-like facial reactions and evaluate the increased similarity to the prototypical pain-like face evoked by mechanical stimuli. The assessments were made one day before and four days after a unilateral mental nerve compression. Our findings show that mental nerve injury promotes an increase in spontaneous facial pain-like expressions and reduced mechanical threshold, reflected in a higher similarity to our pain-like face prototype, regardless of the intensity of the stimuli applied. Machine vision encodes the facial expression associated with evoked and spontaneous pain after mental nerve injury for up to four days. Facial expression quantitatively reflects the increased mechanical sensitivity elicited by mental nerve injury. We also show that this technique can detect spontaneous pain-like responses from facial reactions. Artificial vision can be applied to evaluate signs of orofacial neuropathic pain to study the involved neural circuits.


A single-ward approach to the elderly mentally ill.
A service for the elderly mentally ill is described which is based in a general hospital complex and combines a heavy community commitment with the use of two all-purpose inpatient wards and two associated day hospitals. Each ward functions as an admission, treatment, rehabilitation, and medium- and long-stay facility. Experience suggests that this system makes it possible to cater for the elderly mentally ill in a population with many fewer beds than the recommended norm and is a better insurance against unnecessary prolonged hospitalization than "progressive patient care".


Interpretation of Data Workshop in the Pediatric Preclerkship Educational Exercises (PRECEDE) Curriculum.
The PRECEDE (preclerkship educational exercises) curriculum was developed by the Johns Hopkins Pediatrics Clerkship with the primary goal of providing students with experiential, explicit, and standardized instruction in essential pediatric clinical skills to better prepare them for their clerkships. This data interpretation workshop is one of a series of modules within the PRECEDE curriculum presented just prior to the clerkship. This 2-hour workshop divides medical students into small groups of four to six students, each group with a faculty facilitator. Three clinical vignettes are presented to the student groups. For each vignette, students participate in a guided discussion to develop an interpretation and understanding of what is often misinterpreted pediatric information, such as developmental milestones, vital signs, and laboratory values. From the perspectives of students and instructors, the implementation of this new module was very successful in terms of its delivery of educational content and its organizational structure. Of 229 students who participated in the session and completed surveys, 77% strongly agreed the session was educationally valuable. By establishing important basic skills of correct pediatric data interpretation, students may be better equipped to develop appropriate differential diagnoses, assessments, and plans of care for patients.


Mosquito metabolomics reveal that dengue virus replication requires phospholipid reconfiguration via the remodeling cycle.
Dengue virus (DENV) subdues cell membranes for its cellular cycle by reconfiguring phospholipids in humans and mosquitoes. Here, we determined how and why DENV reconfigures phospholipids in the mosquito vector. By inhibiting and activating the de novo phospholipid biosynthesis, we demonstrated the antiviral impact of de novo-produced phospholipids. In line with the virus hijacking lipids for its benefit, metabolomics analyses indicated that DENV actively inhibited the de novo phospholipid pathway and instead triggered phospholipid remodeling. We demonstrated the early induction of remodeling during infection by using isotope tracing in mosquito cells. We then confirmed in mosquitoes the antiviral impact of de novo phospholipids by supplementing infectious blood meals with a de novo phospholipid precursor. Eventually, we determined that phospholipid reconfiguration was required for viral genome replication but not for the other steps of the virus cellular cycle. Overall, we now propose that DENV reconfigures phospholipids through the remodeling cycle to modify the endomembrane and facilitate formation of the replication complex. Furthermore, our study identified de novo phospholipid precursor as a blood determinant of DENV human-to-mosquito transmission.


Genetic Regulation of Development in Sorghum bicolor: II. Effect of the ma(3) Allele Mimicked by GA(3).
The class III, ma(3) (R)-containing maturity genotypes of Sorghum bicolor (L.) Moench differ from all of the other maturity genotypes in that they flower earlier, exhibit rapid shoot elongation sooner after floral differentiation, and exhibit several morphological differences. In the present study, a class III, ma(3) (R)-containing genotype (44M) was compared to two other genotypes (60M, 80M) that differed only at the third maturity locus. GA(3) treatment caused 60M and 80M to undergo floral differentiation earlier, to exhibit an increase in shoot elongation rate sooner after floral initiation, and to exhibit morphological changes including: increased leaf blade and leaf sheath length, decreased leaf width and leaf area, as well as increased dry weight, especially in the culm weight. Since every major morphological character which is different in class III genotypes was changed by GA(3) treatment of the class II genotypes (60M, 80M), thereby causing them to be more like the class III genotypes, it is clear that application of GA(3) makes maturity genotypes with Ma(3) or ma(3) behave as if they contained ma(3) (R). All of these results support the hypothesis that the ma(3) (R) allele increases gibberellin activity in the maturity genotypes with that allele.


New pUC-derived cloning vectors with different selectable markers and DNA replication origins.
Four new Escherichia coli cloning vectors are described, pUC6S, pUC21, pUK21 and pOK12. These vectors contain a polylinker or multiple cloning site (MCS) with the recognition sequences for 28 restriction enzymes. Plasmids pUC21, pUK21, and pOK12 contain the MCS in the N-terminal end of the lacZ alpha fragment allowing blue/white screening for inserts. To potentially increase the stability of some inserts that may encode toxic proteins, the strength of the lacZ promoter present on these vectors has been reduced. Plasmids pUC6S and pUC21 carry the bla gene encoding ampicillin resistance, while pUK21 and pOK12 contain the gene encoding kanamycin resistance. Plasmid pOK12 carries the replicon from P15A, resulting in a lower copy number pUC-type vector. Plasmid pUC6S carries the ori and bla gene present on all pUC vectors, but does not contain any lac sequences. Plasmids pUC21 and pUK21 contain the M13 intergenic region allowing for the production of plasmid single-stranded DNA. To improve the yield of ss plasmid DNA, two plasmid cis-acting factors that affect yield were also examined: the effect of plasmid-derived transcription across the M13 ori, and the effect of delecting the M13 minus-strand ori from the plasmid.


Diurnal and circadian rhythmicity of the human blood transcriptome overlaps with organ- and tissue-specific expression of a non-human primate.
Twenty-four-hour rhythmicity in mammalian tissues and organs is driven by local circadian oscillators, systemic factors, the central circadian pacemaker and light-dark cycles. At the physiological level, the neural and endocrine systems synchronise gene expression in peripheral tissues and organs to the 24-h-day cycle, and disruption of such regulation has been shown to lead to pathological conditions. Thus, monitoring rhythmicity in tissues/organs holds promise for circadian medicine; however, most tissues and organs are not easily accessible in humans and alternative approaches to quantify circadian rhythmicity are needed. We investigated the overlap between rhythmic transcripts in human blood and transcripts shown to be rhythmic in 64 tissues/organs of the baboon, how these rhythms are aligned with light-dark cycles and each other, and whether timing of tissue-specific rhythmicity can be predicted from a blood sample. We compared rhythmicity in transcriptomic time series collected from humans and baboons using set logic, circular cross-correlation, circular clustering, functional enrichment analyses, and least squares regression. Of the 759 orthologous genes that were rhythmic in human blood, 652 (86%) were also rhythmic in at least one baboon tissue and most of these genes were associated with basic processes such as transcription and protein homeostasis. In total, 109 (17%) of the 652 overlapping rhythmic genes were reported as rhythmic in only one baboon tissue or organ and several of these genes have tissue/organ-specific functions. The timing of human and baboon rhythmic transcripts displayed prominent 'night' and 'day' clusters, with genes in the dark cluster associated with translation. Alignment between baboon rhythmic transcriptomes and the overlapping human blood transcriptome was significantly closer when light onset, rather than midpoint of light, or end of light period, was used as phase reference point. The timing of overlapping human and baboon rhythmic transcriptomes was significantly correlated in 25 tissue/organs with an average earlier timing of 3.21 h (SD 2.47 h) in human blood. The human blood transcriptome contains sets of rhythmic genes that overlap with rhythmic genes of tissues/organs in baboon. The rhythmic sets vary across tissues/organs, but the timing of most rhythmic genes is similar in human blood and baboon tissues/organs. These results have implications for development of blood transcriptome-based biomarkers for circadian rhythmicity in tissues and organs.


Risk Factors for Complications of Traumatic Injuries.
Complications in hospitalized trauma patients are major causes of morbidity and mortality. The aims of this study were to identify the in-hospital trauma patients' complications and identify the risk factors for complications in this population. A retrospective analysis was conducted in a sample from a Brazilian hospital. The sample consisted of 407 patients, 194 (47.66%) of whom had records of complications. The most common complications were infections (41.80%). The risk factors related to the complications were age, length of hospital stay, external causes, and injury severity. The complications were frequent in this sample, and the risk for complications was characterized by multiple factors.


The effect of dehydroepiandrosterone administration on intestinal calcium absorption in ovariectomized female rats.
Dehydroepiandrosterone (DHEA) administration reportedly recovers osteoporosis, a bone disorder associated with bone deficiency in postmenopausal women. However, the physiological mechanism of DHEA in osteoporosis remains elusive, especially in terms of intestinal calcium absorption. Therefore, we investigated the effect of DHEA administration on calcium absorption in ovariectomized (OVX) female rats using an estrogen receptor antagonist. Female Sprague-Dawley rats (n=23, 6 weeks old) were randomized into three groups: OVX control group (OC, n=7), OVX with DHEA treatment group (OD, n=8), and OVX with DHEA inhibitor group (ODI, n=8) for 8 weeks. Intestinal calcium accumulation, as well as the rate of absorption, demonstrated no significant differences during the experimental period among investigated groups. The bone mineral density (BMD) of the tibia at the proximal metaphysis was higher in the OD group than that in the OC group (p<0.05); however, BMD of the ODI group showed no significant difference from investigated groups. Furthermore, the BMD of the tibia at the diaphysis did not significantly differ among these groups. We revealed that DHEA administration does not involve intestinal Ca absorption, although this treatment improves BMD levels in OVX rats. These observations indicate that the effect of DHEA on the bone in postmenopausal women is solely due to its influence on bone metabolism and not intestinal calcium absorption.


Unusually late-onset mycophenolate mofetil-related colitis.
Serious gastrointestinal complications arising 13 years after the initiation of posttransplant immunosuppressant therapy with mycophenolate mofetil are reported. Over a three-month period, a male heart transplant recipient who had taken oral mycophenolate mofetil (2 g daily) for 13 years as part of an immunosuppressant maintenance regimen developed diarrhea and weight loss leading to renal failure and metabolic acidosis. There was no evidence of opportunistic infection, and immunostaining for cytomegalovirus yielded negative results. Colonoscopy revealed areas of congested, erythematous, and nodular mucosa. Histological examination of mucosal biopsy specimens revealed pathological abnormalities typical of those seen in cases of mycophenolate mofetil-associated colitis. On discontinuation of mycophenolate mofetil use, the patient's diarrhea resolved and his renal function improved. Colitis, diarrhea, and other gastrointestinal complications are commonly reported in patients receiving mycophenolate mofetil, an immunosuppressant widely used to prevent rejection of solid organ or bone marrow transplants; however, the onset of such symptoms after more than a decade of continuous use of the drug has not been previously reported. This case suggests that mycophenolate mofetil toxicity should be considered in the evaluation of late-onset posttransplant diarrhea regardless of the duration of therapy. A 33-year-old man maintained on mycophenolate mofetil for 13 years after heart transplantation developed diarrhea, weight loss, and acute kidney injury over a three-month period. Colonoscopy and biopsy revealed pathological changes consistent with mycophenolate mofetil toxicity, and the patient's symptoms resolved after the drug was discontinued.


Driving down the electronic highway.
Life as a computer lab coordinator is never dull nor routine. No sooner do you become comfortable with computer-aided instruction (CAI), computerized testing, interactive video systems, and local area networks, than do you face a new challenge. As faculty and students become more sophisticated in learning how to use computers, interest in accessing large, international computer networks increases. This article describes the available resources, and both the rewards and problems associated with preparing yourself to teach nursing students about the Internet, a large, global association of government, academic, and research computer networks.


Does outsourcing affect hospital profitability?
Organizations outsource non-core service functions to achieve cost reductions and strategic benefits, both of which can impact profitability performance. This article examines relations between managerial outsourcing decisions and profitability for a multi-state sample of non-profit hospitals, across 16 states and four regions of the United States. Overall regression results indicate that outsourcing does not necessarily improve hospital profitability. In addition, we identify no profitability impact from outsourcing for urban hospitals, but somewhat positive effects for teaching hospitals. Our regional analysis suggests that hospitals located in the Midwest maintain positive profitability effects with outsourcing, but those located in the South realize negative effects. These findings have implications for cost reduction efforts and the financial viability of non-profit hospitals.


Multiphoton Scattering Tomography with Coherent States.
In this work we develop an experimental procedure to interrogate the single- and multiphoton scattering matrices of an unknown quantum system interacting with propagating photons. Our proposal requires coherent state laser or microwave inputs and homodyne detection at the scatterer's output, and provides simultaneous information about multiple-elastic and inelastic-segments of the scattering matrix. The method is resilient to detector noise and its errors can be made arbitrarily small by combining experiments at various laser powers. Finally, we show that the tomography of scattering has to be performed using pulsed lasers to efficiently gather information about the nonlinear processes in the scatterer.


Acute infectious bursal disease in poultry: Isolation and characterisation of a highly virulent strain.
A highly virulent strain of infectious bursal disease virus (IBDV) was isolated from the field and propagated in SPF chickens, causing up to 100% mortality. Although it still belongs to the standard serotype 1 IBD viruses, serological typing with monoclonal antibodies showed an antigenic drift in this pathogenic strain. Conventional 'intermediate' IBD vaccines are probably more antigenically related to the pathogenic strain than the mild ones and were effective in protecting SPF chickens against challenge.


Impact of Nutrient Availability on Tertiary Wastewater Treatment by Chlorella vulgaris.
Algae can use excessive nutrients from wastewater effluent to generate beneficial products such as biofuels. However, fluctuation of wastewater characteristics could hinder the implementation of tertiary algal treatment. This study aims to identify the impact of nutrient availability on Chlorella vulgaris's ability to remove nitrogen and phosphorus from wastewater, and its potential as a biofuel feedstock. Experiments using synthetic wastewater with varying concentrations of nitrate and phosphate showed C. vulgaris continued to remove either nutrient when the other was exhausted. Nitrogen starvation led the algae to accumulate the highest amount of neutral lipid; however, the exhaustion of phosphorus did not produce such impact. Synergistic effect was also observed between C. vulgaris and indigenous microorganisms in nutrient removal from real wastewater effluent. The results showed C. vulgaris can survive in a range of nutrient-limiting conditions, making tertiary algal treatment applicable following various secondary treatment regimes.


Large-scale computational identification of HIV T-cell epitopes.
Bioinformatics-driven T-cell epitope-identification methods can enhance vaccine target selection significantly. We evaluated three unrelated computational methods to screen Pol, Gag and Env sequences extracted from the Los Alamos HIV database for HLA-A*0201 and HLA-B*3501 T-cell epitope candidates. The hidden Markov model predicted 389 HLA-B*3501-restricted candidates from 374 HIV-1 and 97 HIV-2 sequences. The artificial neural network (ANN) model, and Bioinformatics and Molecular Analysis Section (BIMAS) quantitative matrix predictions for A*0201 yielded 1122 HIV-1 and 548 HIV-2 candidates. The overall sequence coverage of the predicted A*0201 T-cell epitopes was 2.7% (HIV-1)and 3.0% (HIV-2). HLA-B*3501-predicted epitopes covered 0.9% (HIV-1) and 1.4% (HIV-2) of the total sequence. Comparison of 890 ANN- and 397 BIMAS-derived HIV-1 A*0201- restricted epitope candidates showed that only 13-19% of the predicted and 26% of the experimentally confirmed T-cell epitopes were captured by both methods. Extrapolating these results, we estimated that at least 247 predicted HIV-1 epitopes are yet to be discovered as active A*0201-restricted T-cell epitopes. Adequate comparison and combined usage of various predictive bioinformatics methods, rather than uncritical use of any single prediction method, will enable cost-effective and efficient T-cell epitope screening.


A Novel Ferroptosis-Related Gene Signature for Prognosis Prediction in Ewing Sarcoma.
Ferroptosis, as a form of programmed cell death independent of apoptosis, has been demonstrated that plays a major role in tumorigenesis and cancer treatment. A comprehensive analysis of ferroptosis-related genes (FRGs) may lead to a novel choice for the treatment of Ewing sarcoma (ES). Here, 148 differentially expressed FRGs (DEFRGs) were identified between normal and ES tissue. And the GO and KEGG analyses of DEFRGs indicated that these genes were enriched in cancer and immune-related signaling pathways. Then, the GSE17679 cohort was randomly divided into train and test cohorts. Based on the train cohort, AURKA, RGS4, and RIPK1 were identified as key genes through the univariate Cox regression analysis, the random survival forest algorithm, and the multivariate Cox regression analysis and utilized to establish a prognostic FRG signature. The validation results demonstrated that the gene signature has not only excellent prediction performance and generalization ability but is also good at predicting the response of immunotherapy and chemotherapy. Subsequent analysis indicated that all 3 key genes play key roles in tumor immunity and prognosis of ES. Of these, AURKA was highly associated with EWSR1, which was verified by a single-cell dataset (GSE130019). Therefore, the 3 genes may be potential therapeutic targets for ES. At the end of this study, we also constructed an accurate nomogram that helps clinicians to assess the survival time of ES patients. In conclusion, our study constructed an excellent gene signature, which is helpful in improving the prognosis of ES patients.


Blood culture-guided de-escalation of empirical antimicrobial regimen for critical patients in an online antimicrobial stewardship programme.
A blood culture-guided review strategy was applied to a hospital-wide computerised antimicrobial approval system (HCAAS) at a medical centre in Taiwan. The study aimed to evaluate the impact of this deployment on prescribers' behaviours, antimicrobial consumption, antimicrobial expenditure and healthcare quality in adult intensive care units (ICUs). The HCAAS automatically identifies patients with positive blood cultures and notifies the pre-assigned infectious diseases (ID) physicians for an online second review of the current antimicrobial regimen. Patients from 16 adult ICUs were selected as a focus group. Descriptive analysis, McNemar's test, interrupted time-series analysis and univariate regression analysis were applied. The number of prescriptions assigned for second review increased from 304 in 2010 to 682 in 2012. The approval rate for the antimicrobial regimen in the second review exceeded 70%. In disapproved cases, prescribers accepted the recommendation from ID physicians in 66.1% of cases in the first year; the acceptance rate increased to 80.6% in 2012. Among the restricted antimicrobial agents, consumption gradients decreased for all eight drug classes. The overall antimicrobial expenditure gradient declined significantly following deployment of the second review strategy. The healthcare-associated infection rate continued to decrease over time, and the mortality and ICU re-admission rates remained stable after deployment. A blood culture-guided review of antimicrobial use based on clinical and microbiological evidence improves accuracy in choosing appropriate antimicrobial agents and encourages de-escalation. Consumption and expenditure gradients of antimicrobial agents decreased after the intervention, and healthcare quality was not compromised.


[Comparison of Two Anti-HIV Confirmatory Assays: Recombinant HIV 1/2 Line Immunoassay and Geenius HIV 1/2 Confirmatory Test].
The main purpose in the diagnosis of human immunodeficiency virus (HIV) infection is to rapidly and accurately identify people with HIV infection. It is recommended that samples that are repeatedly reactive should be verified/supported according to the classical algorithms of international and national guidelines. The recombinant "line immunoassay test (LIA)", which has been used for many years, is studied with the accumulated samples to be cost and labor-effective. In this study, the supplemental recombinant HIV 1/2 LIA (INNO-LIA®, Fujirebio, Ghent, Belgium) used to confirm and differentiate the diagnosis of HIV-1 and HIV-2 infections, and an immunochromatographic supplemental test (Geenius™ (Bio-Rad Laboratories, Marnes-la-Coquette, France) which can provide faster results were compared. One hundred fifty serum samples sent to Ege University Faculty of Medicine Hospital Medical Virology Laboratory with anti-HIV 1/2 and p24 antigen positive and indeterminant results and three HIV-1 positive external quality control samples were included in the study. Samples were tested both with the Geenius™ HIV 1/2 (Bio-Rad Laboratories, Marnes-la-Coquette, France) and recombinant HIV 1/2 LIA (INNO-LIA®, Fujirebio, Ghent, Belgium). HIV 1 viral load was evaluated by using Abbott real-time HIV-1 test in Abbott m200sp system (Abbott Molecular, Wiesbaden, Germany) in plasma samples. In both assays, the results were consistent in 147 samples (96.08%). Six samples that have discordant results were as follows: one sample was LIA HIV-1 positive and Geenius indeterminate, two samples were LIA indeterminant and Geenius HIV-1 positive, and in three samples, LIA was indeterminate and Geenius negative. In two EIA reactive samples (2/97, 2.06%) and three EIA negative samples (3/53, 5.66%) LIA results were indeterminant. Geenius test, on the other hand, correctly identified HIV positive and negative samples. The immunochromatographic test could be used in the diagnostic algorithm of HIV infection, due to its short application time, not being labor intensive, its ability to distinguish HIV-1/2, its high sensitivity/specificity compared to LIA, and the compliance with LIA. However, it should be noted that in acute HIV infection, all analytical antibody tests, become reactive later than the fourth generation enzyme immunoassays.


The cytogenetics of a Triticum turgidum x Psathyrostachys juncea hybrid and its backcross derivatives.
Psathyrostachys juncea (2n = 2x = 14, NN), a source of barley yellow dwarf (BYDV) virus resistance with tolerance to drought and salinity, has been successfully hybridized in its autotetraploid form (2n = 4x = 28, NNNN) as the pollen parent to durum wheat (Triticum turgidum L.). The 2n = 4x = 28 (ABNN) F1 hybrid has a mean meiotic metaphase-I configuration of 20.29 univalents + 0.29 ring bivalents + 3.36 rod bivalents + 0.14 trivalents. Spike length, internode length, glume awn length and lemma awn length, as well as the general spike morphology of the F1 hybrid, are intermediate with those of the two parents. Pollinating the ABNN F1 hybrid has given backcross (BC)-I derivatives of an amphiploid (AABBNN) that expresses limited self-fertility. BC-2 derivatives have been obtained from these plants. Direct transfers of useful genes from Ps. juncea to wheat would require substantial genetic manipulation strategies. Both conventional and novel methodologies, which may complement each other, and so facilitate reaching an agricultural objective end point, are addressed.