Transplanted corneal stromal cells in vitreous reproduce extracellular matrix of healing corneal stroma.
To characterize the extracellular matrix (ECM) formed by corneal stromal cells after injection into the vitreous. This will provide a basis for future studies on the function of corneal ECM macromolecules. Cell line from rabbit dermal fibroblasts (RAB9) and primary cultures of rabbit corneal stroma fibroblasts (NRCF) were grown to confluence. For each cell type, approximately 1 x 10(6) cells suspended in basal medium were injected into the vitreous of normal rabbits and observed periodically with a slit lamp. After 1,2, and 4 weeks, eyes were processed for transmission electron microscopy (TEM), immunohistochemistry, immunocytochemistry, and in situ hybridization. All cells showed gradual growth within the vitreous along the needle track. Occurrence of retinal detachment and inflammation was variable. Transmission electron microscopy of NRCF confirmed the deposition of ECM reminiscent of the organization of normal fetal corneal stroma. Similar matrices were produced by RAB9. NRCF deposited collagen fibrils similar in diameter to those seen in normal developing and healing corneal stroma. RAB9 produced collagen fibrils with larger diameters. NRCF-transplanted cells synthesized proteoglycans and collagen immunologically identical to decorin proteins and type VI collagen, indicating that the expression of specific ECM is maintained after transplantation. In addition, in situ hybridization showed that type XII collagen mRNA is synthesized by transplanted NRCF similar to healing corneas. Corneal stromal cells transplanted into vitreous produce a matrix morphologically and biochemically similar to that in healing corneal stroma.


Fulminant hepatitis after infliximab in a patient with hepatitis B virus treated for an adult onset still's disease.
Infliximab, a chimeric anti-tumor necrosis factor-alpha monoclonal antibody, has been demonstrated to be efficient and safe in patients with active rheumatoid arthritis and in the management of severe bouts of Crohn's disease. However, the safety of infliximab has not been evaluated in patients infected with hepatitis B virus. We report the case of a 28-year-old woman, with a positive hepatitis B virus surface antigen, who developed fulminant hepatitis 2 weeks after receiving a second infliximab infusion for a refractory adult onset Still's disease.


Immunocytochemical localization of laminin in hamster tracheal epithelial cell cultures.
EM examination of 28 day cultures of enzymatically dissociated hamster tracheal epithelial (HTE) cells grown on collagen coated millipore filters reveals that fragments of basal lamina may be present at the basal plasmalemma. Since the basal lamina consists of several major components including type IV collagen, heparan sulfate proteoglycans, entactin/nidogen, and laminin, questions naturally arise concerning the presence of such a structure in this cell culture system. When immunocytochemical procedures utilizing anti-laminin antibody and PAP techniques are carried out with paraffin sections of HTE culture at 1,2,3, and 4 weeks in vitro, LM analysis reveals that a thin, dense line of reaction product is present between the basal surface of the HTE cells and the underlying collagen substrate. Immunoblotting evaluation carried out with supernatants of 7d HTE cell homogenates and HTE cell conditioned media also indicate that laminin is being produced by the tracheal cells. Thus, the presence of basal lamina-like fragments, the immunocytochemical localization of laminin, and immunoblot identification of laminin in hamster tracheal epithelial cell cultures, suggest that, although basal lamina components may be produced by HTE cells, at the time points tested, they are not yet being organized into a complete basal lamina.


Carotid atherosclerotic plaque segmentation in multi-weighted MRI using a two-stage neural network: advantages of training with high-resolution imaging and histology.
A reliable and automated method to segment and classify carotid artery atherosclerotic plaque components is needed to efficiently analyze multi-weighted magnetic resonance (MR) images to allow their integration into patient risk assessment for ischemic stroke. Certain plaque components such as lipid-rich necrotic core (LRNC) with hemorrhage suggest a greater likelihood of plaque rupture and stroke event. Assessment for presence and extent of LRNC could assist in directing treatment with impact upon patient outcomes. To address the need to accurately determine the presence and extent of plaque components on carotid plaque MRI, we proposed a two-staged deep-learning-based approach that consists of a convolutional neural network (CNN), followed by a Bayesian neural network (BNN). The rationale for the two-stage network approach is to account for the class imbalance of vessel wall and background by providing an attention mask to the BNN. A unique feature of the network training was to use ground truth defined by both high-resolution ex vivo MRI data and histopathology. More specifically, standard resolution 1.5 T in vivo MR image sets with corresponding high resolution 3.0 T ex vivo MR image sets and histopathology image sets were used to define ground-truth segmentations. Of these, data from 7 patients was used for training and from the remaining two was used for testing the proposed method. Next, to evaluate the generalizability of the method, we tested the method with an additional standard resolution 3.0 T in vivo data set of 23 patients obtained from a different scanner. Our results show that the proposed method yielded accurate segmentation of carotid atherosclerotic plaque and outperforms not only manual segmentation by trained readers, who did not have access to the ex vivo or histopathology data, but also three state-of-the-art deep-learning-based segmentation methods. Further, the proposed approach outperformed a strategy where the ground truth was generated without access to the high resolution ex vivo MRI and histopathology. The accurate performance of this method was also observed in the additional 23-patient dataset from a different scanner. In conclusion, the proposed method provides a mechanism to perform accurate segmentation of the carotid atherosclerotic plaque in multi-weighted MRI. Further, our study shows the advantages of using high-resolution imaging and histology to define ground truth for training deep-learning-based segmentation methods.


Prehospital use in emergency patients of a laryngeal mask airway by ambulance paramedics is a safe and effective alternative for endotracheal intubation.
In Dutch ambulance practice, failure or inability to intubate patients with altered oxygenation and/or ventilation leaves bag-valve mask ventilation as the only alternative, which is undesirable for patient outcome. A novel Laryngeal Mask Airway Supreme (LMA-S) device may be a suitable alternative. To evaluate the effectiveness and suitability of the LMA-S for emergency medical services in daily out-of-hospital emergency practice. After a period of theoretical and practical training of ambulance paramedics in the use of the LMA-S, prospective data were collected on the utilisation of LMA-S in an observational study. Procedures for use were standardised and the evaluation included the number of direct intubation attempts before using LMA-S, attempts required, failure rate and the adequacy of ventilation. Data were analysed taking patient characteristics such as age and indication for ventilatory support into account. The LMA-S was used 50 times over a period of 9 months (33 involving cardiorespiratory arrest, 14 primary and three rescue). The LMA-S could be applied successfully in all 50 cases (100%) and was successful in the first attempt in 49 patients (98%). Respiratory parameters showed adequate oxygenation. All paramedics were unanimously positive about the utilisation of LMA-S because of the easiness of the effort of insertion and general use, and emphasised its value as a useful resource for patients in need. Ensuring ventilation support by using LMA-S by paramedics in prehospital emergency practice is safe and effective.


Neuroprotective effect of l-theanine in a rat model of chronic constriction injury of sciatic nerve-induced neuropathic pain.
We investigated the protective efficacy of l-theanine (LT), the major amino acid components of green tea, on chronic constriction injury (CCI) of sciatic nerve-induced neuropathic pain (NP) development and neuronal functional changes in rats. Rats with NP induced by CCI of the left sciatic nerve and sham-operated rats received LT or saline solution, with pain sensitive tests of thermal hyperalgesia and mechanical allodynia. Motor and sensory nerve conduction velocities were measured after surgery. Subsequently, the rats were sacrificed; the sciatic nerve was excised, homogenized, prepared and subjected for estimation of nitric oxide (NO), malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), myeloperoxidase (MPO), and caspase-3. CCI produced a significant increase in hyperalgesia and allodynia, an increase in SFI, a decrease in nerve conduction velocity, increases in NO, MDA, TNF-α, IL-1β, IL-6, MPO, and caspase-3 levels, as well as reduction of GSH, SOD, and CAT in the rat sciatic nerve. LT treatment significantly and dose-dependently alleviated CCI-induced nociceptive pain thresholds and ameliorated abnormal nerve conduction and functional loss in rats with CCI. Moreover, LT treatment reduced NO and MDA levels, increased antioxidative strength, and markedly suppressed the levels of neuroinflammatory and apoptotic markers in injured sciatic nerves. This is the first report on the ameliorative effect of LT in CCI-induced NP in rats. This effect might be attributed to its anti-oxidative, anti-inflammatory, anti-apoptotic, and neuroprotective, thus making it potentially useful as an adjuvant to conventional treatment.


[Standard chest x-ray: from film to all digital!].
Conventional imaging will be totally replaced by digital imaging because of the lower exposure dosage and the clear advantages in terms of image processing, transfer and storage. Reports in the literature have demonstrated that a 2.5 pl/mm spatial resolution is satisfactory to detect a pneumothorax, a parenchymal nodule or a mediastinal mass. For most authors, this resolution is also sufficient to detect an interstitial syndrome. Digitalization by amplification or by particle detectors using wire chambers are not, for the time being, acceptable for interpretation of a simple chest image because of the low spatial resolution. The selenium cylinder is an adapted technology dedicated to chest imaging. Memory display screens can be used to digitalize the entire radiology unit. They are perfectly adapted for bedside imaging. Flat detectors are based on direct or indirect conversion of the X photons into an electrical signal during the clinical evaluation. The early results appear most promising.


Tuning multi/pluri-potent stem cell fate by electrospun poly(L-lactic acid)-calcium-deficient hydroxyapatite nanocomposite mats.
In this study, we investigated whether multipotent (human-bone-marrow-derived mesenchymal stem cells [hBM-MSCs]) and pluripotent stem cells (murine-induced pluripotent stem cells [iPSCs] and murine embryonic stem cells [ESCs]) respond to nanocomposite fibrous mats of poly(L-lactic acid) (PLLA) loaded with 1 or 8 wt % of calcium-deficient nanohydroxyapatite (d-HAp). Remarkably, the dispersion of different amounts of d-HAp to PLLA produced a set of materials (PLLA/d-HAp) with similar architectures and tunable mechanical properties. After 3 weeks of culture in the absence of soluble osteogenic factors, we observed the expression of osteogenic markers, including the deposition of bone matrix proteins, in multi/pluripotent cells only grown on PLLA/d-HAp nanocomposites, whereas the osteogenic differentiation was absent on stem-cell-neat PLLA cultures. Interestingly, this phenomenon was confined only in hBM-MSCs, murine iPSCs, and ESCs grown on direct contact with the PLLA/d-HAp mats. Altogether, these results indicate that the osteogenic differentiation effect of these electrospun PLLA/d-HAp nanocomposites was independent of the stem cell type and highlight the direct interaction of stem cell-polymeric nanocomposite and the mechanical properties acquired by the PLLA/d-HAp nanocomposites as key steps for the differentiation process.


Perspectives on salvage therapy for non-small-cell lung cancer.
Platinum-based chemotherapy offers a modest survival advantage over best supportive care in chemotherapy-naive patients with a good performance status and advanced/metastatic non-small-cell lung cancer (NSCLC). Despite the survival benefit associated with first-line chemotherapy, the majority of patients will experience relapse or disease progression. In clinicalpractice, an increasing number of patients maintain a good performance status after first-line treatment and are eligible for further treatments. Docetaxel (Taxotere) at 75 mg/m2 given once every 3 weeks has been the standard of care for second-line chemotherapy since the year 2000. Pemetrexed (Alimta) is a novel multitargeted antifolate agent with single-agent activity in first- and second-line treatment of NSCLC. A large phase 111 study comparing docetaxel to pemetrexed in second-line therapy demonstrated that pemetrexed is equally active and less toxic than docetaxel. Based on these results, pemetrexed is a reasonable second-line chemotherapy option for patients with recurrent, advanced NSCLC. Progress made in the field of molecular biology has led to the identification of drugs active against specific cellular targets. Gefitinib (Iressa) and erlotinib (Tarceva) are both orally active tyrosine kinase inhibitors of the epidermal growth factor receptor. Phase II and III trials have demonstrated that these agents are active particularly in a subgroup of patients with specific biologic characteristics. Both drugs have been approved for the treatment of pretreated NSCLC. Other drugs, such as cetuximab (Erbitux) and bevacizumab (Avastin) have shown promising activity in NSCLC and are currently being tested in clinical trials.


Cocaine-induced peroxidative stress in rat liver: antioxidant enzymes and mitochondria.
This study investigated how the cocaine-induced lipid peroxidation affected liver antioxidant enzymes and mitochondria. Acute cocaine (40 mg/kg) injection produced a significant, time-dependent increase in manganese-superoxide dismutase (Mn/SOD) activity and cellular thiobarbuturic acid reactive substances (TBARS), but activities of glutathione peroxidase and catalase were reduced significantly. These changes coincided with increased production of reactive oxygen species by mitochondria and decreased cellular ATP and glutathione. Binge cocaine (25 mg/kg; 5 injections in 3 days) significantly increased TBARS and conjugated dienes, but decreased ATP and glutathione. Accumulation of TBARS and reduction of glutathione was seen in mitochondria. Activities and mRNA of Mn/SOD and copper-zinc-superoxide dismutase were significantly elevated, but mRNA and activities of glutathione peroxidase and catalase were decreased in cocaine-treated rats. Cocaine (binge model) produced scattered liver necrosis (20%) and compromised cell integrity. This is the first report demonstrating cocaine-induced liver necrosis in rats. Pretreatment (acute model) with dimethylaminoethyl-2,2-diphenylvalerate, inhibitor of cocaine bioactivation, curtailed in part the generation of reactive oxygen species by mitochondrial fraction. Dimethylaminoethyl-2,2-diphenylvalerate also prevented the increase of TBARS and Mn/SOD. The results suggest that elevated levels of Mn/SOD and copper-zinc-superoxide dismutase, without a concomitant increase in glutathione peroxidase, catalase and glutathione s-transferase, may have contributed to cocaine-induced cellular and mitochondrial peroxidative stress. Reactive metabolites of cocaine N-oxidative metabolism may be responsible for the cocaine-induced oxidative stress and liver necrosis.


Immune evasion as a pathogenic mechanism of varicella zoster virus.
Varicella zoster virus (VZV) is a human herpesvirus that causes varicella (chickenpox) during primary infection, establishes latency in dorsal root ganglia and may reactivate years later, producing herpes zoster. VZV must evade antiviral immunity during three important stages of viral pathogenesis, including the cell-associated viremia characteristic of primary infection, persistence in dorsal root ganglia during latency and the initial period of VZV reactivation. Our observations about the immunomodulatory effects of VZV document its capacity to interfere with adaptive immunity mediated by CD4 as well as CD8 T cells, ensuring the survival of the virus in the human population from generation to generation.


Staphylococcus aureus: colonizing features and influence of an antibacterial treatment in adults with atopic dermatitis.
The skin of up to 100% of patients with atopic dermatitis (AD) is colonized with Staphylococcus aureus. Of all S. aureus strains isolated from lesional skin, up to 65% have been shown to produce exotoxins with superantigenic properties. Patients colonized with S. aureus have been treated with antibiotics in several open and double-blind placebo-controlled studies, with conflicting results. These studies did not consider the anterior nares as a reservoir of S. aureus, or the possibility of transmission between patients and their contacts. Moreover, adult patients have not so far been investigated. To investigate the colonizing features of S. aureus in adults with AD and in their contacts, and the effect of an antimicrobial treatment of the patients and their partners. Swabs were taken from the skin and anterior nares of 66 adults with AD. S. aureus strains were screened for the production of exotoxins in 32 patients. Ten patients (two with toxigenic strains, eight with non-toxigenic strains) were treated orally with cefalexin, chlorhexidine ointment was applied to the skin, and the anterior nares were treated with mupirocin ointment. A bath containing potassium permanganate was taken daily. In addition, their partners were treated topically. Sixty-two of 66 patients (94%) were carriers of S. aureus, and mostly harboured the bacteria on both skin and anterior nares. Ten of 32 (31%) patients were colonized with toxigenic strains. The Severity Scoring in AD (SCORAD) score decreased in nine of 10 patients who received antimicrobial treatment (P < 0.001), and this effect was more pronounced in patients with a baseline SCORAD > 50. S. aureus may play an important role as an aggravating factor in adults with AD, as antimicrobial treatment leads to a significant improvement of AD in patients who are colonized with the bacterium.


Report of a study group on ablate and pace therapy for paroxysmal atrial fibrillation. Barcelona Discussion Group. Working Group on Arrhythmias of the European Society of Cardiology.
Atrioventricular junctional (AVJ) catheter ablation followed by pacemaker implantation is now widely accepted for patients affected by paroxysmal atrial fibrillation (PAF) not controlled by antiarrhythmic drugs. However, few data exist on its indications, optimal methodology and complications. Therefore a study group examined current practice in Europe and North America, using a questionnaire, followed by a Study Group Meeting to discuss the results. Based upon this, class I, class II and class III indications were proposed. Class I indications (for which general agreement existed) include drug-refractory PAF, correlating with important symptoms, the bradycardia tachycardia syndrome already treated with a pacemaker, and continued PAF. Large differences exist in the current methodology, but consensus was reached on the technical approaches of right and left-sided AVJ ablation, and on the timing of pacemaker implant in relation to ablation. No complete agreement was reached on technical features such as catheter choice and heparin use. The recommended pacing mode was DDDR with mode switching.


Theoretical study of the alpha-cyclodextrin dimer.
The molecular structure, stabilization energy, and thermodynamic properties of the plausible modes of the interaction for the three possible alpha-cyclodextrin (alpha-CD) dimers (head-to-head, tail-to-tail, and head-to-tail) with a water cluster were obtained using quantum chemical methods for the first time. Nine distinct spatial arrangements were investigated. The head-to-head mode of interaction with water is preferred by more than 10 kcal.mol(-1) (BLYP/6-31G(d,p)//PM3 Gibbs free energy difference value at room temperature) in relation to the next stable structure, with a water dimer structure placed inside each cavity and cyclic water tetramers surrounding each tail end. The inter alpha-CD hydrogen bonds play a major role to stabilize the dimeric structures, with no water tetramer being found between the two alpha-CD subunits for the preferred global minimum structure. Therefore, a theoretical model aimed to describe the behavior of alpha-CD dimer, or their inclusion complexes, in the aqueous media should take into account this preference for binding of the water molecules.


Impact of an inpatient anticoagulation management service on clinical outcomes.
Antithrombotic medications require careful management to avoid thrombotic or hemorrhagic complications. The benefits of specialized anticoagulation management services (AMS) in the outpatient setting are well established; less evidence of benefit in the hospital setting is available. To evaluate the clinical benefits of an inpatient AMS to cardiac surgery patients requiring warfarin anticoagulation therapy. After obtaining institutional review board approval, we conducted a retrospective, single-center, cohort study of consecutive cardiac surgery patients treated before (January 2003-May 2005) and after (June-December 2005) establishment of an inpatient AMS. Demographic and clinical characteristics as well as laboratory and clinical data were retrieved from institutional electronic databases and compared between the 2 patient cohorts. Comparisons between study groups were conducted using a chi(2) or Fisher's Exact test for categorical variables and a Student's t-test for continuous variables. Analysis of rare event data was conducted using Poisson regression analysis. Of 1919 patients admitted during the study interval, 826 received warfarin (674 pre-AMS, 152 post-AMS). The number of patients with postsurgical panic international normalized ratio (INR) values declined after initiation of the AMS (pre-AMS 90/674 [13.4%] vs post-AMS 11/152 [7.2%]; p = 0.036). There was a trend toward fewer clinically significant postoperative bleeding events (pre-AMS 21/674 [3.1%] vs post-AMS 2/152 [1.3%]; p = 0.22) and fewer repeat surgeries for late postoperative bleeding (pre-AMS 8/674 [1.2%] vs post-AMS 0/152 [0%]; p = 0.08). AMS intervention was associated with a 17% decrease in the average postsurgical length of stay (13.9 days vs 11.6 days; p = 0.015). A multidisciplinary AMS can improve anticoagulation management, leading to fewer panic INR values and a reduced length of hospital stay.


[Investigations into the incidence of the various causative agents of mastitis in cattle. Studies done in the ambulatory clinic of the Faculty of Veterinary Medicine, Utrecht].
The clinical and bacteriological findings in every head of cattle treated for clinical mastitis in the Ambulatory Clinic of the Veterinary Faculty of Utrecht University were recorded over a period of twelve months. Cattle affected with mastitis were classified into a number of groups, a classification based on the clinical symptoms shown by the animals. In each of these groups the incidence of the various causative agents of mastitis was determined. Another subject of study was the distribution of the causative agents over the various groups. The high proportion (34 percent) of coliform micro-organisms, isolated from animals affected with acute mastitis and showing symptoms of generalized disease was a striking feature.


Variants in the CD36 gene associate with the metabolic syndrome and high-density lipoprotein cholesterol.
A region along chromosome 7q was recently linked to components of the metabolic syndrome (MetS) in several genome-wide linkage studies. Within this region, the CD36 gene, which encodes a membrane receptor for long-chain fatty acids and lipoproteins, is a potentially important candidate. CD36 has been documented to play an important role in fatty acid metabolism in vivo and subsequently may be involved in the etiology of the MetS. The protein also impacts survival to malaria and the influence of natural selection has resulted in high CD36 genetic variability in populations of African descent. We evaluated 36 tag SNPs across CD36 in the HyperGen population sample of 2020 African-Americans for impact on the MetS and its quantitative traits. Five SNPs associated with increased odds for the MetS [P = 0.0027-0.03, odds ratio (OR) = 1.3-1.4]. Coding SNP, rs3211938, previously shown to influence malaria susceptibility, is documented to result in CD36 deficiency in a homozygous subject. This SNP conferred protection against the MetS (P = 0.0012, OR = 0.61, 95%CI: 0.46-0.82), increased high-density lipoprotein cholesterol, HDL-C (P = 0.00018) and decreased triglycerides (P = 0.0059). Fifteen additional SNPs associated with HDL-C (P = 0.0028-0.044). We conclude that CD36 variants may impact MetS pathophysiology and HDL metabolism, both predictors of the risk of heart disease and type 2 diabetes.


Prenatal Risk and Infant Regulation: Indirect Pathways via Fetal Growth and Maternal Prenatal Stress and Anger.
Pathways from maternal tobacco, marijuana, stress, and anger in pregnancy to infant reactivity and regulation (RR) at 9 months of infant age were examined in a low-income, diverse sample beginning in the first trimester of pregnancy, with fetal growth and postnatal stress/anger as potential mediators, and infant sex as a moderator. Participants were 247 dyads (173 substance-exposed infants). There were no direct effects of prenatal risk on RR and no moderation by sex. However, there were significant indirect effects on RR via poor fetal growth and higher postnatal anger. The study adds to the sparse literature on joint effects of tobacco and marijuana, and highlights the role of fetal growth and maternal anger as important pathways from prenatal risk to infant RR.


Identification of some cleavage sites of insulin by insulin proteinase.
In a previous study [Muir, Offord & Davies (1986) Biochem. J. 237, 631-637] the chromatographic and electrophoretic behaviour of a major labelled fragment in the degradation of tritiated insulins by insulin proteinase were used to locate the probable sites of cleavage which had produced this fragment. In order to define these cleavage sites more precisely, authentic markers for the fragments which would be produced by cleavages at, or adjacent to, the most likely sites have now been synthesized. These markers were compared with labelled fragments of the A- and B-chains of insulin produced by insulin proteinase. The results, together with those of our previous study, show that in order to produce the observed major labelled fragment, the enzyme must have cleaved the insulin A-chain between leucine-A13 and tyrosine-A14 and the insulin B-chain between serine-B9 and histidine-B10. In addition, a minor component was observed in the labelled B-chain fragment which corresponded to a cleavage either between histidine-B10 and leucine-B11 or between leucine-B11 and valine-B12.


Characterization of initiated cells in N-methylnitrosourea-induced carcinogenesis of the CNS in the adult rat.
Glial tumors may originate from the malignant transformation of multipotent glial progenitor cells, but tools to study malignant transformation leading to gliomas are limited by the lack of biological systems that represent early stages of this disease in adult animals. In order to characterize the initiated cells that give rise to gliomas, we have employed the N-methylnitrosourea (MNU) model for induction of brain tumors in adult rats (Rushing et al., 1998). Specifically, we have isolated and cultured transformed (premalignant) cells from normal-appearing brains of rats exposed to MNU for 10 weeks and from histologically abnormal brains of rats exposed to MNU for 15 weeks. We compared them with cells cultured from control animals under identical conditions. Cultured cells were classified according to their morphology, immunophenotype, karyotype, proliferation capacity, and tumorigenicity in athymic mice. Cultures from untreated normal rat brains grew as monolayers and had normal karyotypes (42 X,Y), epithelioid morphology, and slow proliferative capacity (doubling time > 120 h). In contrast, cultured cells from brains of MNU-exposed animals had karyotypes that ranged from normal to highly aneuploid. Aneuploid lines grew rapidly in multilayers (doubling time < 24 h), had differentiated astrocytic or oligodendroglial morphology and immunohistochemical staining profile, and yielded tumors in athymic mice. Initiated cells with minor chromosomal aberrations assumed mixed bipolar or tripolar morphologies in high density cultures, proliferated rapidly, but showed contact inhibition and failed to induce tumors when injected s.c. in athymic mice. In general, lines showing no evidence of chromosomal aberrations had the most epithelioid morphology, proliferated slowly (doubling time > 72 h), and retained strict contact growth inhibition. The presumed undifferentiated glial progenitor cells in culture from either control or MNU-treated rats variably expressed markers such as vimentin, nestin, and NG2 proteoglycan, and they weakly expressed the mature astrocytic or oligodendroglial markers glial fibrillary acidic protein or galactocerbroside, respectively. These cultures differentiated to bipolar-tripolar morphology with concomitant maturation to a GFAP+ or GalC+ phenotype upon exposure to secondary messengers such as dibutyryl-cyclic-AMP and/or growth factors such as basic fibrillary growth factor. Continuous stimulation with these messengers resulted in terminal differentiation and consequent death upon withdrawal of the stimulus. These results provide information that could lead to detailed characterization of initiated, premalignant cells in the adult brain and to a better understanding of glial carcinogenesis.


Three-dimensional critical behavior with 2D, 1D, and 0D dimensionality crossover: surface and edge specific heats.
The critical behavior at a second order phase transition is characterized by the divergence of the correlation length xi. We have studied the superfluid transition of 4He in a series of experimental cells in which this divergence of xi is modified due to finite-size confinement. In particular, the design of these cells is such that the smallest dimension is kept the same, 1 microm, but the geometry is such that one obtains crossover to dimensionality of 2, 1, and 0. This corresponds to films, channels, and boxes filled with helium. We measure the specific heat and compare these results with theoretical expectations. We identify surface and line specific heat contributions by analyzing the deviation of the specific heat from its behavior in the thermodynamic limit. The design of these cells is made possible by a combination of silicon lithography and direct wafer bonding.


Cytoplasmic dynein mediates adenovirus binding to microtubules.
During infection, adenovirus (Ad) capsids undergo microtubule-dependent retrograde transport as part of a program of vectorial transport of the viral genome to the nucleus. The microtubule-associated molecular motor, cytoplasmic dynein, has been implicated in the retrograde movement of Ad. We hypothesized that cytoplasmic dynein constituted the primary mode of association of Ad with microtubules. To evaluate this hypothesis, an Ad-microtubule binding assay was established in which microtubules were polymerized with taxol, combined with Ad in the presence or absence of microtubule-associated proteins (MAPs), and centrifuged through a glycerol cushion. The addition of purified bovine brain MAPs increased the fraction of Ad in the microtubule pellet from 17.3% +/- 3.5% to 80.7% +/- 3.8% (P < 0.01). In the absence of tubulin polymerization or in the presence of high salt, no Ad was found in the pellet. Ad binding to microtubules was not enhanced by bovine brain MAPs enriched for tau protein or by the addition of bovine serum albumin. Enhanced Ad-microtubule binding was also observed by using a fraction of MAPs purified from lung A549 epithelial cell lysate which contained cytoplasmic dynein. Ad-microtubule interaction was sensitive to the addition of ATP, a hallmark of cytoplasmic dynein-dependent microtubule interactions. Immunodepletion of cytoplasmic dynein from the A549 cell lysate abolished the MAP-enhanced Ad-microtubule binding. The interaction of Ad with both dynein and dynactin complexes was demonstrated by coimmunoprecipitation. Partially uncoated capsids isolated from cells 40 min after infection also exhibited microtubule binding. In summary, the primary mode of Ad attachment to microtubules occurs though cytoplasmic dynein-mediated binding.


Exploring polysaccharide and protein-enriched decellularized matrix scaffolds for tendon and ligament repair: A review.
Tissue engineering (TE) has become a primary research topic for the treatment of diseased or damaged tendon/ligament (T/L) tissue. T/L injuries pose a severe clinical burden worldwide, necessitating the development of effective strategies for T/L repair and tissue regeneration. TE has emerged as a promising strategy for restoring T/L function using decellularized extracellular matrix (dECM)-based scaffolds. dECM scaffolds have gained significant prominence because of their native structure, relatively high bioactivity, low immunogenicity, and ability to function as scaffolds for cell attachment, proliferation, and differentiation, which are difficult to imitate using synthetic materials. Here, we review the recent advances and possible future prospects for the advancement of dECM scaffolds for T/L tissue regeneration. We focus on crucial scaffold properties and functions, as well as various engineering strategies employed for biomaterial design in T/L regeneration. dECM provides both the physical and mechanical microenvironments required by cells to survive and proliferate. Various decellularization methods and sources of allogeneic and xenogeneic dECM in T/L repair and regeneration are critically discussed. Additionally, dECM hydrogels, bio-inks in 3D bioprinting, and nanofibers are briefly explored. Understanding the opportunities and challenges associated with dECM-based scaffold development is crucial for advancing T/L repairs in tissue engineering and regenerative medicine.


Dynamic fracture of nonglassy suspensions.
We study the dynamic fracture of thin layers of suspensions of non-Brownian rigid particles. The impact of a projectile triggers a liquid-to-solid transition and a hole opens in the layer. We show that the occurrence of fracture and the spatial and dynamic features of the cracks depend mostly on the thickness of the layer and the particle volume fraction. In contrast, the properties of the fractured material seem independent of volume fraction. Finally, we measure the velocity of the crack tip, from which we estimate an effective value of the shear modulus of the fractured material.


Hallucinogenic plant poisoning in children.
Datura is a hallucinogenic plant found in urban or rural areas in the Kingdom of Saudi Arabia KSA. It grows wildly in many parts of the country. Its taste and shape makes it unattractive to both man and animals, though deliberate use by young adults for its hallucinogenic effects have been widely reported for the past 30 years. Datura contains 3 main toxic alkaloids: atropine, scopolamine and hyoscamine. Consumption of any part of the plant can result in severe anticholinergic toxicity. Clinical symptoms are those seen in atropine poisoning, particularly mydriasis and hallucinations. Children have a special susceptibility to atropine toxicity; even small amount may produce central nervous system manifestations. Hospitalization is required for agitation and combative behavior although symptomatic treatment is usually sufficient. We report a case of acute Datura stramonium intoxication in a 6-year-old boy from Khamis Mushayt, KSA, who presented with restlessness, hallucinations and mydriasis 8 hours after ingesting the seeds of Datura plant.


Inflamm-aging of the stem cell niche: breast cancer as a paradigmatic example: breakdown of the multi-shell cytokine network fuels cancer in aged people.
Inflamm-aging is a relatively new terminology used to describe the age-related increase in the systemic pro-inflammatory status of humans. Here, we represent inflamm-aging as a breakdown in the multi-shell cytokine network, in which stem cells and stromal fibroblasts (referred to as the stem cell niche) become pro-inflammatory cytokine over-expressing cells due to the accumulation of DNA damage. Inflamm-aging self-propagates owing to the capability of pro-inflammatory cytokines to ignite the DNA-damage response in other cells surrounding DNA-damaged cells. Macrophages, the major cellular player in inflamm-aging, amplify the phenomenon, by broadcasting pro-inflammatory signals at both local and systemic levels. On the basis of this, we propose that inflamm-aging is a major contributor to the increase in cancer incidence and progression in aged people. Breast cancer will be presented as a paradigmatic example for this relationship.


A Passive RF Testbed for Human Posture Classification in FM Radio Bands.
This paper explores the opportunities and challenges for classifying human posture in indoor scenarios by analyzing the Frequency-Modulated (FM) radio broadcasting signal received at multiple locations. More specifically, we present a passive RF testbed operating in FM radio bands, which allows experimentation with innovative human posture classification techniques. After introducing the details of the proposed testbed, we describe a simple methodology to detect and classify human posture. The methodology includes a detailed study of feature engineering and the assumption of three traditional classification techniques. The implementation of the proposed methodology in software-defined radio devices allows an evaluation of the testbed's capability to classify human posture in real time. The evaluation results presented in this paper confirm that the accuracy of the classification can be approximately 90%, showing the effectiveness of the proposed testbed and its potential to support the development of future innovative classification techniques by only sensing FM bands in a passive mode.


Cytomegalovirus Latency and Reactivation: An Intricate Interplay With the Host Immune Response.
CMV is an ancient herpesvirus that has co-evolved with its host over millions of years. The 236 kbp genome encodes at least 165 genes, four non-coding RNAs and 14 miRNAs. Of the protein-coding genes, 43-44 are core replication genes common to all herpesviruses, while ~30 are unique to betaherpesviruses. Many CMV genes are involved in evading detection by the host immune response, and others have roles in cell tropism. CMV replicates systemically, and thus, has adapted to various biological niches within the host. Different biological niches may place competing demands on the virus, such that genes that are favorable in some contexts are unfavorable in others. The outcome of infection is dependent on the cell type. In fibroblasts, the virus replicates lytically to produce infectious virus. In other cell types, such as myeloid progenitor cells, there is an initial burst of lytic gene expression, which is subsequently silenced through epigenetic repression, leading to establishment of latency. Latently infected monocytes disseminate the virus to various organs. Latency is established through cell type specific mechanisms of transcriptional silencing. In contrast, reactivation is triggered through pathways activated by inflammation, infection, and injury that are common to many cell types, as well as differentiation of myeloid cells to dendritic cells. Thus, CMV has evolved a complex relationship with the host immune response, in which it exploits cell type specific mechanisms of gene regulation to establish latency and to disseminate infection systemically, and also uses the inflammatory response to infection as an early warning system which allows the virus to escape from situations in which its survival is threatened, either by cellular damage or infection of the host with another pathogen. Spontaneous reactivation induced by cellular aging/damage may explain why extensive expression of lytic genes has been observed in recent studies using highly sensitive transcriptome analyses of cells from latently infected individuals. Recent studies with animal models highlight the potential for harnessing the host immune response to blunt cellular injury induced by organ transplantation, and thus, prevent reactivation of CMV and its sequelae.


Optical computing and investigations on writing.
In previous years several methods have been developed for extracting significant information parameters contained in written, typed, or printed texts. They have shown that some models (optical, mathematical, or hybrid) could be described for linear and nonlinear operations. At first, essential features of information contained in a written page are quantitatively specified and related to legibility criteria. Then an approach to handwriting analysis is made through coherent optical processing along two distinct ways: (1) averaging permits assessing a degree of resemblance between any letter in a set and that taken as a reference, (2) as similarities cannot be extracted directly, the introduction of a form function applies to the comparison of printed letters of various types taken as a whole and other kinds of graphic signs. Finally a nonlinear model built up from polynomials can afford a complete analysis of handwriting as illustrated by various examples.


Cancer patterns among children of Turkish descent in Germany: a study at the German Childhood Cancer Registry.
Cancer risks of migrants might differ from risks of the indigenous population due to differences in socioeconomic status, life style, or genetic factors. The aim of this study was to investigate cancer patterns among children of Turkish descent in Germany. We identified cases with Turkish names (as a proxy of Turkish descent) among the 37,259 cases of childhood cancer registered in the German Childhood Cancer Registry (GCCR) during 1980-2005. As it is not possible to obtain reference population data for children of Turkish descent, the distribution of cancer diagnoses was compared between cases of Turkish descent and all remaining (mainly German) cases in the registry, using proportional cancer incidence ratios (PCIRs). The overall distribution of cancer diagnoses was similar in the two groups. The PCIRs in three diagnosis groups were increased for cases of Turkish descent: acute non-lymphocytic leukaemia (PCIR 1.23; CI (95%) 1.02-1.47), Hodgkin's disease (1.34; 1.13-1.59) and Non-Hodgkin/Burkitt lymphoma (1.19; 1.02-1.39). Age, sex, and period of diagnosis showed no influence on the distribution of diagnoses. No major differences were found in cancer patterns among cases of Turkish descent compared to all other cases in the GCCR. Slightly higher proportions of systemic malignant diseases indicate that analytical studies involving migrants may help investigating the causes of such cancers.


Involuntary treatment in patients with anorexia nervosa: utilization patterns and associated factors.
A subgroup of patients with anorexia nervosa (AN) undergoing involuntary treatment (IT) seems to account for most of the IT events. Little is known about these patients and their treatment including the temporal distribution of IT events and factors associated with subsequent utilization of IT. Hence, this study explores (1) utilization patterns of IT events, and (2) factors associated with subsequent utilization of IT in patients with AN. In this nationwide Danish register-based retrospective exploratory cohort study patients were identified from their first (index) hospital admission with an AN diagnosis and followed up for 5 years. We explored data on IT events including estimated yearly and total 5-year rates, and factors associated with subsequent increased IT rates and restraint, using regression analyses and descriptive statistics. IT utilization peaked in the initial few years starting at or following the index admission. A small percentage (1.0%) of patients accounted for 67% of all IT events. The most frequent measures reported were mechanical and physical restraint. Factors associated with subsequent increased IT utilization were female sex, lower age, previous admissions with psychiatric disorders before index admission, and IT related to those admissions. Factors associated with subsequent restraint were lower age, previous admissions with psychiatric disorders, and IT related to these. High IT utilization in a small percentage of individuals with AN is concerning and can lead to adverse treatment experiences. Exploring alternative approaches to treatment that reduce the need for IT is an important focus for future research.


Optical fiber-coupled InGaAs-based terahertz time-domain spectroscopy system.
The successful demonstration of an optical fiber-coupled terahertz time-domain spectroscopy (THz-TDS) system is described in this study. The terahertz output power of the emitter with two optical band rejection filters was 132 nW, which is an improvement of 70% over the output power without any filters. This improvement is due to the suppression of an optical modulated signal that is reverse-generated when an alternating current bias exceeding a certain threshold is applied to the emitter. Under the optimal alignment conditions, the terahertz detector in a fiber-coupled THz-TDS system clearly measured water vapor dips in the free space.


Hypoxia imaging and theranostic potential of [64Cu][Cu(ATSM)] and ionic Cu(II) salts: a review of current evidence and discussion of the retention mechanisms.
Tumor hypoxia (low tissue oxygenation) is an adverse condition of the solid tumor environment, associated with malignant progression, radiotherapy resistance, and poor prognosis. One method to detect tumor hypoxia is by positron emission tomography (PET) with the tracer [64Cu][Cu-diacetyl-bis(N(4)-methylthiosemicarbazone)] ([64Cu][Cu(ATSM)]), as demonstrated in both preclinical and clinical studies. In addition, emerging studies suggest using [64Cu][Cu(ATSM)] for molecular radiotherapy, mainly due to the release of therapeutic Auger electrons from copper-64, making [64Cu][Cu(ATSM)] a "theranostic" agent. However, the radiocopper retention based on a metal-ligand dissociation mechanism under hypoxia has long been controversial. Recent studies using ionic Cu(II) salts as tracers have raised further questions on the original mechanism and proposed a potential role of copper itself in the tracer uptake. We have reviewed the evidence of using the copper radiopharmaceuticals [60/61/62/64Cu][Cu(ATSM)]/ionic copper salts for PET imaging of tumor hypoxia, their possible therapeutic applications, issues related to the metal-ligand dissociation mechanism, and possible explanations of copper trapping based on studies of the copper metabolism under hypoxia. We found that hypoxia selectivity of [64Cu][Cu(ATSM)] has been clearly demonstrated in both preclinical and clinical studies. Preclinical therapeutic studies in mice have also demonstrated promising results, recently reporting significant tumor volume reductions and improved survival in a dose-dependent manner. Cu(II)-[Cu(ATSM)] appears to be accumulated in regions with substantially higher CD133+ expression, a marker for cancer stem cells. This, combined with the reported requirement of copper for activation of the hypoxia inducible factor 1 (HIF-1), provides a possible explanation for the therapeutic effects of [64Cu][Cu(ATSM)]. Comparisons between [64Cu][Cu(ATSM)] and ionic Cu(II) salts have showed similar results in both imaging and therapeutic studies, supporting the argument for the central role of copper itself in the retention mechanism. We found promising evidence of using copper-64 radiopharmaceuticals for both PET imaging and treatment of hypoxic tumors. The Cu(II)-[Cu(ATSM)] retention mechanism remains controversial and future mechanistic studies should be focused on understanding the role of copper itself in the hypoxic tumor metabolism.


Cavity linewidth narrowing and broadening due to competing linear and nonlinear dispersions.
We experimentally demonstrate cavity linewidth control by manipulating dispersion of the intracavity medium. By making use of the dramatic change of Kerr nonlinearity near electromagnetically induced transparency resonance in a three-level atomic system, the cavity transmission linewidth can be greatly modified. As the cavity input intensity increases, the cavity linewidth changes from below to above empty cavity linewidth, corresponding to subluminal and superluminal photon propagation in the cavity, respectively.


Bronchial response to nebulized antibiotics in children with cystic fibrosis.
Nebulized antibiotics are being used increasingly in children with cystic fibrosis. We assessed the effect of nebulized antibiotic solutions of varying tonicity on lung function in 12 children aged 5-15 yrs with cystic fibrosis. Baseline forced expiratory volume in one second and (FEV1) was measured, followed by a single nebulization of normal saline (272 mosmol.kg-1), tobramycin (248 mosmol.kg-1), or ticarcillin (3,080 mosmol.kg-1). All children received each of these, administered randomly, one per day. FEV1 was remeasured 5, 15 and 30 min after completion of the nebulization. Ticarcillin (mean fall 10.7% (SD 8.9)) caused a larger fall in FEV1 than normal saline (4.8% (4.3), p less than 0.05). The fall in FEV1 for ticarcillin was greater than for tobramycin (1.2% (2.0), p less than 0.05). Normal saline did not result in a significantly larger fall in FEV1 than tobramycin (p greater than 0.05). Bronchoconstriction to ticarcillin persisted at 30 min. We conclude that nebulized antibiotics can affect lung function in children with cystic fibrosis if the solutions are hypertonic.


Terminal deoxynucleotidyl transferase-containing cells in peripheral blood: implications for the surveillance of patients with lymphoblastic leukemia or lymphoma in remission.
An indirect immunofluorescence assay was used to quantitate TdT-containing (TdT+) cells in the mononuclear leukocyte fraction of peripheral blood from normal subjects and patients with acute lymphoblastic leukemia (ALL) and lymphoblastic lymphoma (LL). In normal children (10) and adults (10), 0.036% +/- 0.014% (mean +/- SD) and 0.030% +/- 0.015% TdT+ cells were found. In peripheral bloods from 10 children receiving chemotherapy for tumors other than ALL or LL, 0.040% +/- 0.039% TdT+ cells were found. Serial determinations were performed on 15 patients with ALL or LL who were in clinical remission. Eight of these patients remained in continuous remission and always had fewer than 0.11% TdT+ cells in their peripheral blood. Three patients who developed systemic relapse were found to have progressively rising numbers of TdT+ cells in their peripheral blood prior to clinical evidence of relapse. All 3 of these patients had greater than 0.1% TdT+ cells in their peripheral blood from 3 to 8 wk prior to clinical relapse. In 3 other patients, localized extramedullary relapse developed, but no trend was found on serial TdT determinations. Thus, the indirect immunofluorescence assay for TdT detects a small population of cells in normal peripheral blood. In patients with ALL, progressive increases above this normal level were associated with subsequent bone marrow relapse.


Deubiquitinating enzymes AtUBP12 and AtUBP13 and their tobacco homologue NtUBP12 are negative regulators of plant immunity.
• Signalling by ubiquitination is implicated in diverse aspects of the plant lifecycle, and enzymes of ubiquitin metabolism are overrepresented in the Arabidopsis genome compared with other model eukaryotes. Despite the importance of ubiquitination in the regulation of signalling, little is known about deubiquitinating enzymes, which reverse the process of ubiquitination. • Transgenic RNA interference-based cosuppression and the isolation of Atubp12/13 double mutants collectively provides the first report that AtUBP12 and AtUBP13 are functionally redundant and are required for immunity against virulent Pseudomonas syringae pv tomato in Arabidopsis. The Solanaceous AtUBP12 orthologue NtUBP12 was identified. Viral-induced gene silencing and transient gain-of-function assays were employed to establish that the NtUBP12 protein functions as a negative regulator of the Cf-9-triggered hypersensitive response. • Here, we demonstrate that NtUBP12 and AtUBP12 are bona fide deubiquitinating enzymes capable of cleaving lysine-48-linked ubiquitin chains. AtUBP12 and NtUBP12 are functionally interchangeable and their deubiquitinating activity is required to suppress plant cell death. • Overall, our data implicate AtUBP12- and NtUBP12-dependent deubiquitination in the stabilization of common substrates across Solanaceae and Brassicaceae which regulate disease resistance.


Development of a multi-point polarization-based vibration sensor.
In this paper we propose a novel kind of multi-point vibration sensor based on the polarization properties of light. Its principle relies on the combination of mechanical transducers with fiber Bragg gratings. When subject to vibrations, the mechanical transducers induce birefringence variations within the fiber and in turn modify the state of polarization, which appears as a power variation after going through a polarizer. The FBGs reflect light from different positions of the sensing fiber and provide wavelength multiplexing. We show that this sensor can provide the vibration frequencies in a quasi-distributed manner.


Calcitonin-loaded octamaleimic acid-silsesquioxane nanoparticles in hydrogel scaffold support osteoinductivity in bone regeneration.
Novel osteoinductive scaffolds fabricated using the benefits of tissue engineering techniques accompanied by utilizing drugs can accelerate bone regeneration. The purpose of this study was to load salmon calcitonin (sCT) in octamaleimic acid-silsesquioxane (OMA-POSS) nanoparticles and enrich the hydrogel scaffold based on hydroxyapatite, Gelrite® and platelet-rich plasma (PRP) for use in bone tissue engineering. The loading efficiency, release percentage, particle size and zeta potential of the nanoparticles were evaluated. The proliferation of seeded MG-63 osteoblast cells on the designed scaffold, its cytotoxicity and osteo-conductivity were studied by alkaline phosphatase measurement and Alizarin red staining. The expression of cellular osteogenic markers such as collagen 1 (COL1A1), osteocalcin (BGLAP) and osteopontin (SPP1) was examined using reverse transcription polymerase chain reaction. The results revealed that the particle size of the nanoparticles varied between 94.2 and 199.2 nm and their negative surface charge increased after drug conjugation. The osteoblast cell proliferation and calcium granule production in the optimum formulation were significantly higher in comparison with the control group (p < 0.05). Osteogenic markers increased significantly after a specific number of days of cell culture compared to the control group (p < 0.05). The results also showed the potential of the designed scaffold in bone tissue engineering.


Minimizing Isolate Catalyst Motion in Metal-Assisted Chemical Etching for Deep Trenching of Silicon Nanohole Array.
The instability of isolate catalysts during metal-assisted chemical etching is a major hindrance to achieve high aspect ratio structures in the vertical and directional etching of silicon (Si). In this work, we discussed and showed how isolate catalyst motion can be influenced and controlled by the semiconductor doping type and the oxidant concentration ratio. We propose that the triggering event in deviating isolate catalyst motion is brought about by unequal etch rates across the isolate catalyst. This triggering event is indirectly affected by the oxidant concentration ratio through the etching rates. While the triggering events are stochastic, the doping concentration of silicon offers a good control in minimizing isolate catalyst motion. The doping concentration affects the porosity at the etching front, and this directly affects the van der Waals (vdWs) forces between the metal catalyst and Si during etching. A reduction in the vdWs forces resulted in a lower bending torque that can prevent the straying of the isolate catalyst from its directional etching, in the event of unequal etch rates. The key understandings in isolate catalyst motion derived from this work allowed us to demonstrate the fabrication of large area and uniformly ordered sub-500 nm nanoholes array with an unprecedented high aspect ratio of ∼12.


SiMA: A simplified migration assay for analyzing neutrophil migration.
In lung inflammation, neutrophils are the first leukocytes migrating to an inflammatory site, eliminating pathogens by multiple mechanisms. The term "migration" describes several stages of neutrophil movement to reach the site of inflammation, of which the passage of the interstitium and basal membrane of the airway are necessary to reach the site of bronchial inflammation. Currently, several methods exist (e.g., Boyden Chamber, under-agarose assay, or microfluidic systems) to assess neutrophil mobility. However, these methods do not allow for parameterization on single cell level, that is, the individual neutrophil pathway analysis is still considered challenging. This study sought to develop a simplified yet flexible method to monitor and quantify neutrophil chemotaxis by utilizing commercially available tissue culture hardware, simple video microscopic equipment and highly standardized tracking. A chemotaxis 3D µ-slide (IBIDI) was used with different chemoattractants [interleukin-8 (IL-8), fMLP, and Leukotriene B4 (LTB4 )] to attract neutrophils in different matrices like Fibronectin (FN) or human placental matrix. Migration was recorded for 60 min using phase contrast microscopy with an EVOS® FL Cell Imaging System. The images were normalized and texture based image segmentation was used to generate neutrophil trajectories. Based on these spatio-temporal information a comprehensive parameter set is extracted from each time series describing the neutrophils motility, including velocity and directness and neutrophil chemotaxis. To characterize the latter one, a sector analysis was employed enabling the quantification of the neutrophils response to the chemoattractant. Using this hard- and software framework we were able to identify typical migration profiles of the chemoattractants IL-8, fMLP, and LTB4 , the effect of the matrices FN versus HEM as well as the response to different medications (Prednisolone). Additionally, a comparison of four asthmatic and three non-asthmatic patients gives a first hint to the capability of SiMA assay in the context of migration based diagnostics. Using SiMA we were able to identify typical migration profiles of the chemoattractants IL-8, fMLP, and LTB4 , the effect of the matrices FN versus HEM as well as the response to different medications, that is, Prednisolone induced a change of direction of migrating neutrophils in FN but no such effect was observed in human placental matrix. In addition, neutrophils of asthmatic individuals showed an increased proportion of cells migrating toward the vehicle. With the SiMA platform we presented a simplified but yet flexible platform for cost-effective tracking and quantification of neutrophil migration. The introduced method is based on a simple microscopic video stage, standardized, commercially available, µ-fluidic migration chambers and automated image analysis, and track validation software. © 2017 International Society for Advancement of Cytometry.


Improved strategies to counter the COVID-19 pandemic: Lockdowns vs. primary and community healthcare.
COVID-19 pandemic mitigation strategies are mainly based on social distancing measures and healthcare system reinforcement. However, many countries in Europe and elsewhere implemented strict, horizontal lockdowns because of extensive viral spread in the community which challenges the capacity of the healthcare systems. However, strict lockdowns have various untintended adverse social, economic and health effects, which have yet to be fully elucidated, and have not been considered in models examining the effects of various mitigation measures. Unlike commonly suggested, the dilemma is not about health vs wealth because the economic devastation of long-lasting lockdowns will definitely have adverse health effects in the population. Furthermore, they cannot provide a lasting solution in pandemic containment, potentially resulting in a vicious cycle of consecutive lockdowns with in-between breaks. Hospital preparedness has been the main strategy used by governments. However, a major characteristic of the COVID-19 pandemic is the rapid viral transmission in populations with no immunity. Thus, even the best hospital system could not cope with the demand. Primary, community and home care are the only viable strategies that could achieve the goal of pandemic mitigation. We present the case example of Greece, a country which followed a strategy focused on hospital preparedness but failed to reinforce primary and community care. This, along with strategic mistakes in epidemiological surveillance, resulted in Greece implementing a second strict, horizontal lockdown and having one of the highest COVID-19 death rates in Europe during the second wave. We provide recommendations for measures that will reinstate primary and community care at the forefront in managing the current public health crisis by protecting hospitals from unnecessary admissions, providing primary and secondary prevention services in relation to COVID-19 and maintaining population health through treatment of non-COVID-19 conditions. This, together with more selective social distancing measures (instead of horizontal lockdowns), represents the only viable and realistic long-term strategy for COVID-19 pandemic mitigation.


A novel niosome formulation for encapsulation of anthocyanins and modelling intestinal transport.
The bioavailability of drugs can be improved by regulating the structural properties, particularly lipoid systems, such as niosomes, can increase cellular uptake. Herein, we optimized double emulsion and niosomal formulations for encapsulating anthocyanin-rich black carrot extract. Nanoparticles obtained by selected formulation were characterized in terms of morphology, particle size, drug encapsulation efficiency, in vitro release and cytotoxicity. The optimum conditions for niosomal formulation were elicited as 30 mg of cholesterol, 150 mg of Tween 20 and feeding time of 1 min at a stirring rate of 900 rpm yielding the lowest average particle size of 130 nm. In vitro release data showed the majority of the encapsulated anthocyanins were released at the end of 10 h. A mathematical model was developed to estimate the absorption of anthocyanins released from niosomes and cytotoxicity was assessed against neuroblastoma. Overall, these findings suggest that niosomal vesicles might be suitable delivery systems for anthocyanins.


The polyepitope approach to DNA vaccination.
The identification of the epitope as the smallest immunogenic subunit derived from antigenic proteins has promoted the development of epitope-based vaccines. These prevent the danger of administering whole proteins or genes that have unknown and possibly dangerous properties. Recent data suggest that DNA encoded epitopes in synthetic constructs can be processed and presented to CD8+ T-lymphocytes despite unnatural flanking amino acid sequences. This has allowed the development of polyepitope vaccines for cancer and infectious disease which induce multiple cytotoxic T-lymphocyte responses. The resultant immunity may be restricted to various HLA alleles and targeted to numerous antigens to avoid escape from immune detection by antigen loss variants. This review will describe the background of epitope and polyepitope-based vaccination strategies and the more recent results that confirm the potential of DNA encoded polyepitope vaccines. Future directions that may aid the design of more effective polyepitopes will also be discussed.


Lipid Transporters Beam Signals from Cell Membranes.
Lipid composition in cellular membranes plays an important role in maintaining the structural integrity of cells and in regulating cellular signaling that controls functions of both membrane-anchored and cytoplasmic proteins. ATP-dependent ABC and P4-ATPase lipid transporters, two integral membrane proteins, are known to contribute to lipid translocation across the lipid bilayers on the cellular membranes. In this review, we will highlight current knowledge about the role of cholesterol and phospholipids of cellular membranes in regulating cell signaling and how lipid transporters participate this process.


The status and future directions of comprehensive tobacco control policies for the 2020 Tokyo Olympic Games: A review.
This review aims to clarify the status and future directions of comprehensive tobacco control policies for the 2020 Tokyo Olympic Games based on a comparison with seventeen countries that hosted the Olympic Games. Seventeen countries that hosted the Olympic Games from 1988 to 2018 were identified by searching the website of IOC. A comparison of the contents was carried out by six themes (Monitor, Protect, Offer, Warn, Enforcement and Raise) in accordance with the MPOWER package for implementation of the FCTC. The comparison items and data were obtained from Global Health Observatory (GHO) data, Japanese government websites, and the literature. Based on the MPOWER scale, the 17 countries were ranked by total score, and a correlation between smoking prevalence and MPOWER total scores was established. The following three results were clarified: 1) Compared to the 17 countries that hosted the Olympic Games, smoking prevalence in Japan at 22.5% is at a moderate level and male smoking prevalence (33.7%) is still higher than other developed countries such as UK (24.7%) and Australia (16.5%); 2) Japanese tobacco control policies were far behind other countries that hosted the Olympic Games, especially in protecting people from tobacco smoke, warning about the dangers of tobacco, anti-tobacco mass media campaigns and enforcing bans on tobacco advertising, promotion, and sponsorship; and 3) The better practice in MPOWER was able to reduce total and male smoking prevalence in the study countries. Japanese tobacco control policies are far behind other countries that hosted the Olympic Games. In order to successfully hold the 2020 Tokyo Olympic Games, it will be necessary to strengthen tobacco control policies, based on the FCTC, in the future.


Structure and function of plasmid pColD157 of enterohemorrhagic Escherichia coli O157 and its distribution among strains from patients with diarrhea and hemolytic-uremic syndrome.
In this study, pColD157, a 6.7-kb colicinogenic plasmid of enterohemorrhagic Escherichia coli (EHEC) O157:H7 strain CL40cu, was characterized by restriction mapping and determination of its complete nucleotide sequence. The sequence consists of 6,675 bp and shows a high degree of similarity to the nucleotide sequence of colicinogenic plasmids pColD-CA23 and pColK. Seven potential genes were located on pColD157, three of which were closely related (>97.9%) to the colicin D structural gene and the corresponding immunity and lysis genes of plasmid pColD-CA23, and these were therefore designated cda, cdi, and cdl, respectively, using the reference extension -CL40 for differentiation. The adjacent 3' region is related to the origin of replication of pColD-CA23. In contrast, the remaining part of the plasmid harbors a cluster of genes, closely related to the mobilization genes of pColK, which is followed by a 0.3-kb stretch homologous to the pColK resolution function. These determinants were designated mbdA, mbdB, mbdC, and mbdD and cdr, respectively. Southern blot analysis was performed with a probe specific for the cda gene of pColD157 and two groups of EHEC O157:H7 isolates from patients with diarrhea or hemolytic-uremic syndrome resident in Germany. Whereas 16 of 46 E. coli O157 strains isolated between 1987 and 1991 harbored plasmid pColD157, only 1 of 50 strains isolated during 1996 carried this plasmid. In addition, all strains harboring plasmid pColD157 were shown to have colicinogenic activity.


lncRNA CASC2/miR‑18a‑5p axis regulates the malignant potential of nasopharyngeal carcinoma by targeting RBBP8.
Nasopharyngeal carcinoma (NPC) is a prevalent head and neck tumor which has a high mortality rate in Southeast Asia, especially in Southern China. Cancer susceptibility candidate 2 (CASC2) is a newly identified long non‑coding RNA (lncRNA) that has been found to play a suppressive role in several types of tumors. However, the expression and functional role of CASC2 in NPC are still unclear. In the present study, using NPC tissues, cells and transplanted mice, we investigated the mechanism of CASC2‑mediated regulation of NPC. We showed that the CASC2 level is reduced in NPC tissues and cells. CASC2 downregulation promoted proliferation and inhibited apoptotic cell death in NPC cells. In contrast, CASC2 upregulation inhibited proliferation and increased apoptosis. There were putative binding sites of microRNA (miR)‑18a‑5p in the promoter of CASC2. The level of miR‑18a‑5p was upregulated in NPC tissues and cells. We further confirmed that CASC2 could directly bind with miR‑18a‑5p and inhibit miR‑18a‑5p expression, using reporter gene and RNA immunoprecipitation assays. miR‑18a‑5p suppressed CASC2 upregulation‑mediated decrease in proliferation and increase in apoptotic cell death. Bioinformatics predicted the putative binding site of miR‑18a‑5p in the 3' untranslated region of C‑terminal binding protein interacting protein (CtIP)/RBBP8. It was further confirmed that miR‑18a‑5p could directly bind with RBBP8 and inhibit RBBP8 expression. Downregulation of RBBP8 inhibited the anti‑miR‑18a‑5p‑mediated increase in apoptosis and decrease in proliferation. Downregulation of CASC2 increased tumor growth, increased the level of miR‑18a‑5p and decreased RBBP8 expression in vivo. In summary, CASC2 regulates NPC malignancy through modulation of RBBP8 via sponging miR‑18a‑5p. Our findings highlight the CASC2/miR‑18a‑5p/RBBP8 axis in NPC pathogenesis and provide new biomarkers and potential targets for the therapy of NPC.


[Sonography of adrenal gland diseases in the newborn infant].
Diseases and morphological changes of the adrenal gland play an important role especially in the newborn, because malformations, congenital errors of metabolism, connatal tumours and birth injuries become symptomatic in this period of life. In this paper, the possibilities of neonatal adrenal ultrasonography for diagnosis and in the aspect of differential diagnosis are discussed. The characteristic sonographic appearance is the thin echogenic core surrounded by a thick transonic zone. Adrenal haemorrhage as well as the neuroblastoma disturb these structures and show their own characteristic pattern. In congenital adrenal hyperplasia the organ is enlarged in a fold-like manner. In agenesis or displacement of the kidney the adrenal gland presents as an elongated mass in the renal fossa and on the psoas muscle.


Cloning and assisted reproductive techniques: influence on early development and adult phenotype.
Over the past 40 years, our increased understanding and development of cell and molecular biology has allowed even greater advances in reproductive biology. This is most evident by the development of various aspects of assisted reproductive techniques (ART), generation of transgenic animals, and most recently generation of mammals through somatic cell cloning. To date, cloning from adult somatic cells has been successful in at least 10 mammalian species. Although generating viable cloned mammals from adult cells is technically feasible and the list of successes will only continue to grow with time, prenatal and perinatal mortality is high and live cloned offspring have not been without health problems. The success of many of the proposed applications of the cloning technique obviously depends upon the health and survival of founder animals generated by nuclear transfer. This article summarizes the health consequences of cloning in mice, and discusses possible mechanisms through which these conditions may arise. In addition, we discuss the effects of ART in animal models and in humans. ART also involves some of the same procedures used in cloning, and there are reports that offspring generated by ART sometimes display aberrant phenotypes as well. It is important to point out that although these techniques do sometimes produce abnormalities, the majority of offspring are born apparently normal and survive to adulthood. Additionally, we must emphasize that the effects of ART and cloning observed in animal models do not necessarily indicate that they will occur in humans. In this article, we review studies examining the phenotype of animals generated by cloning and various ART, and discuss clinical implications of these findings.


Structure of native Renilla reinformis luciferin.
The structure of native luciferin from the bioluminescent coelenterate Renilla reniformis is shown to be 3,7-dihydro-2-(p-hydroxybenzyl)-6-(p-hydroxyphenyl)-8-benzylimidazo[1,2-a]pyrazin-3-one by mass spectral analysis of synthetic luciferin and the luciferin derived from a protein directly involved in the bioluminescent system. A previous report of the molecular weight of luciferin is shown to be incorrect by reexamination of the spectral data and by synthesis of two derivatives. Detailed analysis of kinetic, emission, and quantum yield data for the isolated and synthetic luciferins confirms this structure. Confirmation of this structure in a number of species from different phyla suggests a common substrate for a variety of bioluminescent marine organisms.


Digital implementation of hierarchical vector quantization.
A formal methodology drives the design and realization of a digital very large-scale integration (VLSI) device supporting hierarchical vector quantization (HVQ) in computation-intensive coding applications. The hardware-oriented model-selection approach enhances the Minimum Description Length criterion with circuit-related aspects that allow consistent and efficient design. The resulting model parameters drive the subsequent realization in digital circuitry, which has first been implemented in field-programmable gate array (FPGA) technology to verify its correctness. The eventual VLSI realization results in an HVQ chip providing cost-effective, computationally efficient real-time performances. Real-world applications support the consistency of the vector quantization approach and the effectiveness of the HVQ device.


Differential gene expression in small and large rainbow trout derived from two seasonal spawning groups.
Growth in fishes is regulated via many environmental and physiological factors and is shaped by the genetic background of each individual. Previous microarray studies of salmonid growth have examined fish experiencing either muscle wastage or accelerated growth patterns following refeeding, or the influence of growth hormone and transgenesis. This study determines the gene expression profiles of genetically unmanipulated large and small fish from a domesticated salmonid strain reared on a typical feeding regime. Gene expression profiles of white muscle and liver from rainbow trout (Oncorhynchus mykiss) from two seasonal spawning groups (September and December lots) within a single strain were examined when the fish were 15 months of age to assess the influence of season (late fall vs. onset of spring) and body size (large vs. small). Although IGFBP1 gene expression was up-regulated in the livers of small fish in both seasonal lots, few expression differences were detected in the liver overall. Faster growing Dec. fish showed a greater number of differences in white muscle expression compared to Sept. fish. Significant differences in the GO Generic Level 3 categories 'response to external stimulus', 'establishment of localization', and 'response to stress' were detected in white muscle tissue between large and small fish. Larger fish showed up-regulation of cytoskeletal component genes while many genes related to myofibril components of muscle tissue were up-regulated in small fish. Most of the genes up-regulated in large fish within the 'response to stress' category are involved in immunity while in small fish most of these gene functions are related to apoptosis. A higher proportion of genes in white muscle compared to liver showed similar patterns of up- or down-regulation within the same size class across seasons supporting their utility as biomarkers for growth in rainbow trout. Differences between large and small Sept. fish in the 'response to stress' and 'response to external stimulus' categories for white muscle tissue, suggests that smaller fish have a greater inability to handle stress compared to the large fish. Sampling season had a significant impact on the expression of genes related to the growth process in rainbow trout.


Adipocyte-derived cytokine resistin causes endothelial dysfunction of porcine coronary arteries.
Resistin, a novel adipocyte-derived cytokine, is involved in the development of insulin resistance and diabetes mellitus. In this study, we determined whether resistin could affect vasomotor function, oxidative stress, and endothelial nitric oxide synthase (eNOS) expression in porcine coronary arteries. Porcine coronary arteries were treated with resistin or antioxidant seleno-L-methionine (SeMet). Vasomotor function was studied by using a myograph system. Levels of superoxide anion (O 2 - ) were detected by the lucigenin-enhanced chemiluminescence method. The eNOS mRNA and protein levels were determined by real-time polymerase chain reaction and immunohistochemistry, respectively. Culture of isolated porcine coronary artery endothelial cells (PCAECs) was also included. Endothelium-dependent relaxation in response to bradykinin was reduced by 15% and 30% for the rings treated with 10 and 40 ng/mL of resistin, respectively, as compared with controls ( P < .05). Endothelium-independent relaxation in response to sodium nitroprusside (SNP) was also reduced by 11% after treatment with 40 ng/mL of resistin ( P < .05). The O 2 - level was increased in the 40 ng/mL resistin-treated vessels by 88% as compared with controls ( P < .05). SeMet reversed these effects. The eNOS mRNA levels in PCAEC cultures treated with resistin (10 and 40 ng/mL) were decreased by 27% and 55%, respectively ( P < .05) and by 39% in the endothelial cells purified from porcine coronary artery rings after treatment with 40 ng/mL of resistin ( P < .05). Immunoreactivity of eNOS in the resistin-treated vessel rings was also substantially reduced. Resistin reduces the endothelium-dependent and endothelium-independent vasorelaxation. This effect is associated with increased superoxide radical production, decreased eNOS expression, and is effectively reversed by the antioxidant SeMet. Obesity has been considered to be an independent risk factor for coronary artery disease and other vascular lesions. Resistin is a newly discovered adipocyte-derived cytokine, and its plasma levels are increased in obese individuals. However, it is not clear whether resistin could directly contribute to vascular disease formation. This study showed that resistin can cause endothelial dysfunction in porcine coronary arteries through oxidative stress and down-regulation of eNOS. Thus, this study may suggest a new mechanism of obesity-associated vascular disease and that antioxidants may effectively prevent vascular disease in obese individuals.


[Comparative research at the "mesoptometer" and "nyktomat" (author's transl)].
This paper deals with the results of comparative reserach into twilight vision as found by tests at the "Mesoptometer" and "Nyktomat". The research population consisted of 168 persons. Their reactions at the test instruments during various functional tests are demonstrated by means of diagrams. While there is good correspondence between the two test instruments in the range of contrast essential for attesting night driving ability, differences can be shown to exist in the critical range of marginal values.


A comparative study of monoaminergic involvement in the antinociceptive action of E-2078, morphine and U-50,488E.
E-2078 ([N-methyl-Tyr1, N-alpha-methyl-Arg7, D-Leu8]dynorphin A(1-8) ethylamide) is a systemically active dynorphin analog. We examined monoaminergic involvement in the antinociceptive action of E-2078 compared with morphine and U-50,488E (trans-3,4-dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)- benzene-acetamide monohydrochloride). The antinociceptive effect of these drugs was determined using the mouse tail-flick test after depletion of norepinephrine or 5-HT by pretreatment with various neurotoxins. Reserpine (i.p.) depleted both norepinephrine and 5-HT. Selective degeneration of noradrenergic nerves was induced by N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4, i.p.) or intrathecal (i.t.) 6-hydroxydopamine (6-OHDA), whereas 5-HT was depleted by p-chlorophenylalanine (PCPA, i.p.) or 5,7-dihydroxytryptamine (5,7-DHT, i.t.). The antinociceptive action of E-2078 administered s.c. was significantly attenuated in mice treated with reserpine, DSP-4, 6-OHDA, PCPA or 5,7-DHT compared with that in non-neurotoxin animals. Antinociception induced by intracerebroventricular (i.c.v.) and i.t. injection of E-2078 was reversed by reserpine, DSP-4 or PCPA. The antinociceptive action of morphine (s.c.) was attenuated by reserpine, DSP-4, 6-OHDA and PCPA, but not by 5,7-DHT. Antinociception produced by i.c.v. morphine was antagonized by reserpine, DSP-4 and PCPA. In contrast, morphine given i.t. was not affected by these neurotoxins. U-50,488E (s.c.)-induced antinociception was attenuated by reserpine, DSP-4, 6-OHDA, PCPA and 5,7-DHT. Intracerebroventricular U-50,488E was antagonized by reserpine, DSP-4 and PCPA, whereas i.t. U-50,488E was reversed only by reserpine and PCPA(ABSTRACT TRUNCATED AT 250 WORDS)


Experiences of student and trainee autism researchers during the COVID-19 pandemic.
Circumstances surrounding the COVID-19 pandemic have resulted in significant personal and professional adjustments. Students and trainees, including those in autism research, face unique challenges to accomplishing their training and career goals during this unprecedented time. In this commentary, we, as members of the International Society for Autism Research Student and Trainee Committee, describe our personal experiences, which may or may not align with those of other students and trainees. Our experiences have varied both in terms of the ease (or lack thereof) with which we adapted and the degree to which we were supported in the transition to online research and clinical practice. We faced and continue to adjust to uncertainties about future training and academic positions, for which opportunities have been in decline and have subsequently negatively impacted our mental health. Students and trainees' prospects have been particularly impacted compared to more established researchers and faculty. In addition to the challenges we have faced, however, there have also been unexpected benefits in our training during the pandemic, which we describe here. We have learned new coping strategies which, we believe, have served us well. The overarching goal of this commentary is to describe these experiences and strategies in the hope that they will benefit the autism research community moving forward. Here, we provide a set of recommendations for faculty, especially mentors, to support students and trainees as well as strategies for students and trainees to bolster their self-advocacy, both of which we see as crucial for our future careers. LAY SUMMARY: The COVID-19 pandemic has affected students and trainees, including those in autism research, in different ways. Here, we describe our personal experiences. These experiences include challenges. For example, it has been difficult to move from in-person to online work. It has also been difficult to keep up with work and training goals. Moreover, working from home has made it hard to connect with our supervisors and mentors. As a result, many of us have felt unsure about how to make the best career choices. Working in clinical services and getting to know and support our patients online has also been challenging. Overall, the pandemic has made us feel more isolated and some of us have struggled to cope with that. On the other hand, our experiences have also included benefits. For example, by working online, we have been able to join meetings all over the world. Also, the pandemic has pushed us to learn new skills. Those include technical skills but also skills for well-being. Next, we describe our experiences of returning to work. Finally, we give recommendations for trainees and supervisors on how to support each other and to build a strong community.


The development of the tobacco tactics website.
Web-based cessation interventions have been shown to reduce tobacco use, be more efficacious than self-help booklets, be more efficacious if they provide tailored messages, and enhance quit rates in conjunction with nicotine replacement therapy. The objective of this study was to usability test and pilot test the Tobacco Tactics website for veterans. Both formative and summative evaluations were used across three small successive studies to develop and test the Tobacco Tactics website for veterans, which was based on a prior face-to-face smoking cessation intervention. Once the website was developed, the research team and Web developers usability tested the website with 5 veteran smokers and former smokers. Feedback from the veterans was collected as they navigated each webpage, then used to revise the website. In pilot study 1, 9 veteran smokers were provided access to the website, and given a baseline and 30-day follow-up survey. In pilot study 2, 18 veteran smokers, who were also motivated to quit smoking, were recruited and randomized to either the Tobacco Tactics website plus nicotine replacement therapy or to the 1-800-QUIT-NOW telephone line. As a result of usability testing, more than 27 modifications were made to improve the website. In pilot study 1, 50% (3/6) veterans who entered the website had cut down on the number of cigarettes and 83% (5/6) found the website enjoyable, easy to read, easy to navigate, and would recommend the website to others. In pilot study 2, which included only smokers motivated to quit and also offered nicotine replacement therapy, seven-day point prevalence abstinence at 30-day follow-up was 40% (4/10) in the intervention group compared to 13% (1/8) in the control group. These preliminary results are promising and suggest the need for wider-scale testing of the Tobacco Tactics website for veterans.


[Metabolic syndrome: how about bone?].
In the last decades, the association between cardiovascular disease and osteoporosis has been a matter of an extensive clinical and basic investigation, and in fact, common pathogenic links have been recently proposed. The metabolic syndrome includes a series of cardiovascular risk factors that confer a high risk of morbimortality. On the other hand, osteoporosis and its more serious consequence, fracture, represent a true epidemic nowadays. The aim of the present article is to perform a brief update of the association between the individual components of the metabolic syndrome and bone metabolism, and to analyze the publications that have studied the same association, but considering the metabolic syndrome as a whole.


Zwilch, a new component of the ZW10/ROD complex required for kinetochore functions.
The Zeste-White 10 (ZW10) and Rough Deal (ROD) proteins are part of a complex necessary for accurate chromosome segregation. This complex recruits cytoplasmic dynein to the kinetochore and participates in the spindle checkpoint. We used immunoaffinity chromatography and mass spectroscopy to identify the Drosophila proteins in this complex. We found that the complex contains an additional protein we name Zwilch. Zwilch localizes to kinetochores and kinetochore microtubules in a manner identical to ZW10 and ROD. We have also isolated a zwilch mutant, which exhibits the same mitotic phenotypes associated with zw10 and rod mutations: lagging chromosomes at anaphase and precocious sister chromatid separation upon activation of the spindle checkpoint. Zwilch's role within the context of this complex is evolutionarily conserved. The human Zwilch protein (hZwilch) coimmunoprecipitates with hZW10 and hROD from HeLa cell extracts and localizes to the kinetochores at prometaphase. Finally, we discuss immunoaffinity chromatography results that suggest the existence of a weak interaction between the ZW10/ROD/Zwilch complex and the kinesin-like kinetochore component CENP-meta.


Pre-radiotherapy lymphocyte count and platelet-to-lymphocyte ratio may improve survival prediction beyond clinical factors in limited stage small cell lung cancer: model development and validation.
Few small sample size studies have reported lymphocyte count was prognostic for survival in small-cell lung cancer (SCLC). This study aimed to validate this finding, to build prediction model for overall survival (OS) and to study whether novel models that combine lymphocyte-related variables can predict OS more accurately than a conventional model using clinical factors alone in a large cohort of limited-stage SCLC patients. This study enrolled 544 limited-stage SCLC patients receiving definitive chemo-radiation with pre-radiotherapy lymphocyte-related variables including absolute lymphocyte count (ALC), platelet-to-lymphocyte ratio (P/L ratio), neutrophil-to-lymphocyte ratio (N/L ratio), and lymphocyte-to-monocyte ratio (L/M ratio). The primary endpoint was OS. These patients were randomly divided into a training dataset (n=274) and a validation dataset (n=270). Multivariate survival models were built in the training dataset, and the performance of these models were further tested in the validation dataset using the concordance index (C-index). The median follow-up time was 36 months for all patients. In the training dataset, univariate analysis showed that ALC (P=0.020) and P/L ratio (P=0.023) were significantly correlated with OS, while L/M ratio (P=0.091) and N/L ratio (P=0.436) were not. Multivariate modeling demonstrated the significance of ALC (P=0.063) and P/L ratio (P=0.003), and the improvement for OS prediction in combined models with the addition of ALC (C-index =0.693) or P/L ratio (C-index =0.688) over the conventional model (C-index =0.679). The validation dataset analysis confirmed a modest improvement of C-index with the addition of ALC or P/L ratio. All these models showed reasonable discriminations and calibrations. This study validated the significant value of pre-radiotherapy ALC and P/L ratio on OS in limited-stage SCLC. The combined model with ALC or P/L ratio showed additional OS prediction values than the conventional model with clinical factors alone.


Order-disorder (OD) polytypism of K3FeTe2O8(OH)2(H2O)1+x.
K3FeTe2O8(OH)2(H2O)2 was synthesized under hydrothermal conditions from Te(OH)6, FeSO4·7H2O and 85 wt% KOH in a 1:2:6 molar ratio. The crystal structure is built of a triperiodic network. One disordered water molecule per formula unit is located in a channel and can be partially removed by heating. Systematic one-dimensional diffuse scattering indicates a polytypic character, which is best described by application of the order-disorder theory. The major polytype is monoclinic with pseudo-orthorhombic metrics. It is interrupted by fragments of an orthorhombic polytype. The diffraction intensities are analyzed using structure factor calculations.


Purified galactooligosaccharide, derived from a mixture produced by the enzymic activity of Bifidobacterium bifidum, reduces Salmonella enterica serovar Typhimurium adhesion and invasion in vitro and in vivo.
The prebiotic Bimuno(®) is a mixture containing galactooligosaccharides (GOSs), produced by the galactosyltransferase activity of Bifidobacterium bifidum NCIMB 41171 using lactose as the substrate. Previous in vivo and in vitro studies demonstrating the efficacy of Bimuno(®) in reducing Salmonella enterica serovar Typhimurium (S. Typhimurium) colonization did not ascertain whether or not the protective effects could be attributed to the prebiotic component GOS. Here we wished to test the hypothesis that GOS, derived from Bimuno(®), may confer the direct anti-invasive and protective effects of Bimuno(®). In this study the efficacy of Bimuno(®), a basal solution of Bimuno(®) without GOS [which contained glucose, galactose, lactose, maltodextrin and gum arabic in the same relative proportions (w/w) as they are found in Bimuno(®)] and purified GOS to reduce S. Typhimurium adhesion and invasion was assessed using a series of in vitro and in vivo models. The novel use of three dimensionally cultured HT-29-16E cells to study prebiotics in vitro demonstrated that the presence of ∼ 5 mg Bimuno(®) ml(-1) or ∼ 2.5 mg GOS ml(-1) significantly reduced the invasion of S. Typhimurium (SL1344nal(r)) (P<0.0001). Furthermore, ∼ 2.5 mg GOS ml(-1) significantly reduced the adherence of S. Typhimurium (SL1344nal(r)) (P<0.0001). It was demonstrated that cells produced using this system formed multi-layered aggregates of cells that displayed excellent formation of brush borders and tight junctions. In the murine ligated ileal gut loops, the presence of Bimuno(®) or GOS prevented the adherence or invasion of S. Typhimurium to enterocytes, and thus reduced its associated pathology. This protection appeared to correlate with significant reductions in the neutral and acidic mucins detected in goblet cells, possibly as a consequence of stimulating the cells to secrete the mucin into the lumen. In all assays, Bimuno(®) without GOS conferred no such protection, indicating that the basal solution confers no protective effects against S. Typhimurium. Collectively, the studies presented here clearly indicate that the protective effects conferred by Bimuno(®) can be attributed to GOS.


Genetic, structural and clinical analysis of spastic paraplegia 4.
Spastic paraplegia type 4 (SPG4), resulting from heterozygous mutations in the SPAST gene, is the most common form among the heterogeneous group of hereditary spastic paraplegias (HSPs). We aimed to study genetic and clinical characteristics of SPG4 across Canada. The SPAST gene was analyzed in a total of 696 HSP patients from 431 families by either HSP-gene panel sequencing or whole exome sequencing (WES). We used Multiplex ligation-dependent probe amplification to analyze copy number variations (CNVs), and performed in silico structural analysis of selected mutations. Clinical characteristics of patients were assessed, and long-term follow-up was done to study genotype-phenotype correlations. We identified 157 SPG4 patients from 65 families who carried 41 different SPAST mutations, six of which are novel and six are CNVs. We report novel aspects of mutations occurring in Arg499, a case with homozygous mutation, a family with probable compound heterozygous mutations, three patients with de novo mutations, three cases with pathogenic synonymous mutation, co-occurrence of SPG4 and clinically isolated syndrome, and novel or rarely reported signs and symptoms seen in SPG4 patients. Our study demonstrates that SPG4 is a heterogeneous type of HSP, with diverse genetic features and clinical manifestations. In rare cases, biallelic inheritance, de novo mutation, pathogenic synonymous mutations and CNVs should be considered.


Effect of iohexol, metrizamide and ioxaglate on the blood-brain barrier.
The effect upon the blood-brain barrier of the non-ionic contrast medium iohexol, compared with that of ioxaglate and metrizamide, was analysed. The contrast media were injected selectively into the internal carotid arteries of rabbits, and 197Hg Ac2 and Trypan blue were used as markers for evaluation of the injury. Less injury to the blood-brain barrier was caused by iohexol.