Linezolid for tympanostomy tube otorrhea caused by methicillin-resistant Staphylococcus aureus and multiple drug-resistant Streptococcus pneumoniae.
To describe a consecutive series of children with refractory tympanostomy tube otorrhea treated with linezolid and document its clinical effectiveness and adverse effects. Retrospective, single institution case series. The records of children treated with linezolid for refractory gram-positive otorrhea from 2003 to 2007 were analyzed for causative organisms, antimicrobial sensitivities, history of prior medical treatments, time to cessation of otorrhea, adverse effects of linezolid and recurrence of infection. Ten episodes of refractory gram-positive otorrhea treated with linezolid were documented among eight children during the study period. Seven were caused by methicillin-resistant Staphylococcus aureus and three by multiple-drug resistant Streptococcus pneumoniae. Prior treatment regimes included clindamycin, trimethoprim-sulfamethoxazole/rifampin, amoxicillin-clavulanate, and/or a third-generation cephalosporin. Seven of eight children had failed extended courses of fluoroquinolone ear drops. All children were free of otorrhea by the 14th day of twice-daily treatment with oral linezolid at 20mg/kg/day. One child developed two additional episodes of S. aureus otorrhea three and 10 months after initial treatment. Each was successfully controlled with linezolid. There were no adverse effects from linezolid treatment. Oral linezolid is highly effective in the treatment of refractory otorrhea caused by methicillin-resistant S. aureus and multiple drug-resistant S. pneumoniae. It has a low risk of serious side effects. It is one of only a few oral agents with activity against these organisms and is very expensive. Linezolid should be used only when otorrhea cannot be treated with conventional agents or when complications of otitis media pose a risk to life or function.


Fetal posterior cerebral artery in pediatric and adult moyamoya disease: A single-center experience of 480 patients.
In past pediatric and adult cohort studies of moyamoya disease, the fetal posterior cerebral artery has received less attention. Its relationship with the clinical manifestations and collateral circulation of moyamoya disease or ipsilateral cerebral hemispheres remains unclear. We summarize the clinical features of patients with and without fetal posterior cerebral artery moyamoya disease from consecutive cases.We explored the relationship between fetal posterior cerebral arteries and collateral circulation in the ipsilateral cerebral hemispheres, as well as differences among different subgroups of patients.According to the morphology, the fetal posterior cerebral artery is divided into complete fetal posterior cerebral artery and partial fetal posterior cerebral artery. Clinical features were classified as: infarction,hemorrhage,and non-stroke in unilateral/bilateral cerebral hemispheres. Collateral circulation is divided into extracranial vascular compensation and leptomeningeal collateral circulation. Digital subtraction angiography and CT/MR were used to evaluate the blood flow status and clinical characteristics of patients with moyamoya disease. A total of 960 cerebral hemispheres from 142 pediatric patients and 338 adult patients were included in the study. A total of 273 (56.9%) patients had 399 cerebral hemispheres (41.6%) with fetal posterior cerebral arteries. Adults with fetal posterior cerebral arteries had lower rates of infarction (24.6%vs37.3%, P =0.005) and were less likely to have bilateral stroke (8.4%vs11.5%, P =0.038). Cerebral hemispheres with fetal posterior cerebral artery were more likely to have anterior cerebral artery and middle cerebral artery stenosis and less likely to have occlusion (P =0.002, 0.001), and less likely to involve the posterior circulation (P < 0.001). The cerebral hemispheres of the fetal posterior cerebral artery had higher leptomeningeal collateral circulation scores. There are significant differences in extracranial vascular compensation between cerebral hemispheres with and without fetal posterior cerebral artery. Adult patients with fetal posterior cerebral artery were more advanced in Suzuki stage (P =0.017). Our results suggest that fetal posterior cerebral artery is associated with infarct manifestations in pediatric and adult moyamoya disease. In the cerebral hemispheres, the fetal posterior cerebral artery is associated with ipsilateral hemispheric anterior and posterior circulation artery injury, extracranial vascular compensation, leptomeningeal collateral circulation compensation, and infarction. Adult patients with fetal posterior cerebral artery were more advanced in Suzuki stage.


Neurosteroids (allopregnanolone) and alcohol use disorder: From mechanisms to potential pharmacotherapy.
Alcohol Use Disorder (AUD) is a multifaceted relapsing disorder that is commonly comorbid with psychiatric disorders, including anxiety. Alcohol exposure produces a plethora of effects on neurobiology. Currently, therapeutic strategies are limited, and only a few treatments - disulfiram, acamprosate, and naltrexone - are available. Given the complexity of this disorder, there is a great need for the identification of novel targets to develop new pharmacotherapy. The GABAergic system, the primary inhibitory system in the brain, is one of the well-known targets for alcohol and is responsible for the anxiolytic effects of alcohol. Interestingly, GABAergic neurotransmission is fine-tuned by neuroactive steroids that exert a regulatory role on several endocrine systems involved in neuropsychiatric disorders including AUD. Mounting evidence indicates that alcohol alters the biosynthesis of neurosteroids, whereas acute alcohol increases and chronic alcohol decreases allopregnanolone levels. Our recent work highlighted that chronic alcohol-induced changes in neurosteroid levels are mediated by epigenetic modifications, e.g., DNA methylation, affecting key enzymes involved in neurosteroid biosynthesis. These changes were associated with changes in GABAA receptor subunit expression, suggesting an imbalance between excitatory and inhibitory signaling in AUD. This review will recapitulate the role of neurosteroids in the regulation of the neuroendocrine system, highlight their role in the observed allostatic load in AUD, and develop a framework from mechanisms to potential pharmacotherapy.


Multicentric carpotarsal osteolysis is caused by mutations clustering in the amino-terminal transcriptional activation domain of MAFB.
Multicentric carpotarsal osteolysis (MCTO) is a rare skeletal dysplasia characterized by aggressive osteolysis, particularly affecting the carpal and tarsal bones, and is frequently associated with progressive renal failure. Using exome capture and next-generation sequencing in five unrelated simplex cases of MCTO, we identified previously unreported missense mutations clustering within a 51 base pair region of the single exon of MAFB, validated by Sanger sequencing. A further six unrelated simplex cases with MCTO were also heterozygous for previously unreported mutations within this same region, as were affected members of two families with autosomal-dominant MCTO. MAFB encodes a transcription factor that negatively regulates RANKL-induced osteoclastogenesis and is essential for normal renal development. Identification of this gene paves the way for development of novel therapeutic approaches for this crippling disease and provides insight into normal bone and kidney development.


Acetazolamide prevents vacuolar myopathy in skeletal muscle of K(+) -depleted rats.
Acetazolamide and dichlorphenamide are carbonic anhydrase (CA) inhibitors effective in the clinical condition of hypokalemic periodic paralysis (hypoPP). Whether these drugs prevent vacuolar myopathy, which is a pathogenic factor in hypoPP, is unknown. The effects of these drugs on the efflux of lactate from skeletal muscle were also investigated. For 10 days, K(+)-depleted rats, a model of hypoPP, were administered 5.6 mg kg(-1) day(-1) of acetazolamide, dichlorphenamide or bendroflumethiazide (the last is not an inhibitor of CA). Histological analysis of vacuolar myopathy and in vitro lactate efflux measurements were performed in skeletal muscles from treated and untreated K(+)-depleted rats, and also from normokalemic rats. About three times as many vacuoles were found in the type II fibres of tibialis anterioris muscle sections from K(+)-depleted rats as were found in the same muscle from normokalemic rats. In ex vivo experiments, a higher efflux of lactate on in vitro incubation was found in muscles of K(+)-depleted rats compared with that found in muscles from normokalemic rats. After treatment of K(+)-depleted rats with acetazolamide, the numbers of vacuoles in tibialis anterioris muscle decreased to near normal values. Incubation with acetazolamide in vitro inhibited efflux of lactate from muscles of K(+)-depleted rats. In contrast, bendroflumethiazide and dichlorphenamide failed to prevent vacuolar myopathy after treatment in vivo and failed to inhibit lactate efflux in vitro. Acetazolamide prevents vacuolar myopathy in K(+)-depleted rats. This effect was associated with inhibition of lactate transport, rather than inhibition of CA.


Bombesin facilitates GABAergic transmission and depresses epileptiform activity in the entorhinal cortex.
Bombesin and the bombesin-like peptides including neuromedin B (NMB) and gastrin-releasing peptide (GRP) are important neuromodulators in the brain. We studied their effects on GABAergic transmission and epileptiform activity in the entorhinal cortex (EC). Bath application of bombesin concentration-dependently increased both the frequency and amplitude of sIPSCs recorded from the principal neurons in the EC. Application of NMB and GRP exerted the same effects as bombesin. Bombesin had no effects on mIPSCs recorded in the presence of TTX but slightly depressed the evoked IPSCs. Omission of extracellular Ca(2+) or inclusion of voltage-gated Ca(2+) channel blockers, Cd(2+) and Ni(2+), blocked bombesin-induced increases in sIPSCs suggesting that bombesin increases GABA release via facilitating extracellular Ca(2+) influx. Bombesin induced membrane depolarization and slightly increased the input resistance of GABAergic interneurons recorded from layer III of the EC. The action potential firing frequency of the interneurons was also increased by bombesin. Bombesin-mediated depolarization of interneurons was unlikely to be mediated by the opening of a cationic conductance but due to the inhibition of inward rectifier K(+) channels. Bath application of bombesin, NMB and GRP depressed the frequency of the epileptiform activity elicited by deprivation of Mg(2+) from the extracellular solution suggesting that bombesin and the bombesin-like peptides have antiepileptic effects in the brain.


Lymphadenomatous carcinoma of the sublingual gland: report of a first case in an unusual localization.
Lymphadenomatous carcinoma of the salivary gland is a very rare tumor, which hitherto occurred exclusively in the parotid gland. This report describes a case of lymphadenomatous carcinoma of the sublingual gland, which is thought to be the first report of this special entity. A 36-year-old man was seen with slowly growing mass on his right floor of mouth. Upon surgical removal, the mass was well encapsulated. Microscopic examination revealed a biphasic appearance with islands of neoplastic epithelial cells presenting against a dense lymphoid stroma, while the epithelial compartment showed features of malignancy. During 30-months of close follow-up, there has not been any sign of local recurrence or metastasis. The clinical behavior, the age of the patient, and the sublingual localization differentiate this lesion from the sebaceous type, favoring lymphadenomatous carcinoma as a distinct clinical entity.


Global characterization of in vivo enzyme catalytic rates and their correspondence to in vitro kcat measurements.
Turnover numbers, also known as kcat values, are fundamental properties of enzymes. However, kcat data are scarce and measured in vitro, thus may not faithfully represent the in vivo situation. A basic question that awaits elucidation is: how representative are kcat values for the maximal catalytic rates of enzymes in vivo? Here, we harness omics data to calculate kmax(vivo), the observed maximal catalytic rate of an enzyme inside cells. Comparison with kcat values from Escherichia coli, yields a correlation ofr(2)= 0.62 in log scale (p < 10(-10)), with a root mean square difference of 0.54 (3.5-fold in linear scale), indicating that in vivo and in vitro maximal rates generally concur. By accounting for the degree of saturation of enzymes and the backward flux dictated by thermodynamics, we further refine the correspondence between kmax(vivo) and kcat values. The approach we present here characterizes the quantitative relationship between enzymatic catalysis in vitro and in vivo and offers a high-throughput method for extracting enzyme kinetic constants from omics data.


Adenine nucleotide and P-creatine levels in layers of frog retina as a function of dark and light adaptation.
Eight layers of frog retina were analyzed for ATP, P-creatine, ATP + ADP, and AMP under conditions of dark, 2 sec, 2 min, and in the case of AMP, 2 hr of light adaptation. Samples of each layer, usually ranging between 5 and 50 ng, were dissected from lyophilized frozen sections. After brief light exposure, ATP dropped while ADP rose sharply in the pigment epithelium, outer segments, and inner segments; ADP was too low to be measured accurately in the inner retina. The profile of ATP, P-creatine, and ATP + ADP concentrations showed peaks in the inner nuclear and ganglion layers. AMP, by contrast, was highest in the two plexiform layers. Levels in the inner retina dropped after only 2 sec of light but rose after 2 hr to levels that were higher than dark values in all retinal layers. AMP was often characterized by a non-uniform distribution: adjacent areas of a layer agreed very closely in value to each other but could vary several-fold from a different section of the same frog or from another frog exposed to the same conditions. This distribution produced clusters of values, particularly prominent in dark-adapted animals, something not observed with the other metabolites measured. The peaks of AMP in the plexiform layers suggest that AMP may be a by-product of dopamine-stimulated adenylate cyclase which also has peaks in these same layers.


FastGrow: on-the-fly growing and its application to DYRK1A.
Fragment-based drug design is an established routine approach in both experimental and computational spheres. Growing fragment hits into viable ligands has increasingly shifted into the spotlight. FastGrow is an application based on a shape search algorithm that addresses this challenge at high speeds of a few milliseconds per fragment. It further features a pharmacophoric interaction description, ensemble flexibility, as well as geometry optimization to become a fully fledged structure-based modeling tool. All features were evaluated in detail on a previously reported collection of fragment growing scenarios extracted from crystallographic data. FastGrow was also shown to perform competitively versus established docking software. A case study on the DYRK1A kinase, using recently reported new chemotypes, illustrates FastGrow's features in practice and its ability to identify active fragments. FastGrow is freely available to the public as a web server at https://fastgrow.plus/ and is part of the SeeSAR 3D software package.


Obstruction of the major pediatric airway.
1. Do not create an emergency obstruction unless you are prepared to handle it. (It may be best to do nothing until the diagnosis is clear.) 2. Support the patient with oxygen, humidity, and positioning and monitor vital signs. 3. Take as brief, concise, and accurate a history as possible. 4. Determine the anatomic level of obstruction. 5. Estimate the degree of obstruction with the maximal potential hazard. 6. Carry out a physical examination of the nose and neck and, except in suspected epiglottitis, the pharynx, larynx, trachea, and lung. 7. Special x-ray studies--lateral, neck and chest, swallowing function, and inspiratory and expiratory films--are obtained as indicated. 8. Therapeutic support: oxygen, humidity, antibiotics, steroids, and recemic epinephrine. Mechanical support: naso-oral airways, appropriate endotracheal or bronchoscopic tubes. tracheotomy.


Outcomes After Pediatric Out-of-Hospital Cardiopulmonary Interventions.
The aim of the study was to evaluate outcomes after pediatric out-of-hospital cardiopulmonary interventions (CPIs) by emergency medical services (EMS). Children (age, ≤18 years) who received CPI by EMS from 2001 to 2008 were identified from the Utah Department of Health. Cardiopulmonary intervention was defined as oxygenation, ventilation or CPR, and transport to a hospital by EMS. Univariate and multivariable regression analyses evaluated associations between potential predictors and outcomes (death and new neurologic dysfunction). A total of 464 patients (58% male) received EMS attention. For the 71% patients (327) who were alive on EMS arrival, 63% (205) received CPI without CPR. Of note, 6% (12) of these patients died after arrival to the hospital and new neurologic dysfunction was diagnosed in 6% (13). Among the 12 patients who died, 50% (6) were younger than 1 year.On multivariable regression analysis, factors associated with increased risk of death before and in-hospital are the following: age younger than 1 year (odds ratio [OR], 0.26; 95% confidence interval [CI], 0.17-0.39), shorter EMS transport time (OR, 0.94; 95% CI, 0.89-0.99), and longer EMS dispatch time (OR, 1.23; 95% CI, 1.08-1.40). Factors associated with increased risk of new neurologic dysfunction are the following: lack of pulse (OR, 0.14; 95% CI, 0.04-0.53), requiring CPR (OR, 6.15; 95% CI, 1.48-25.6), and CPR duration (OR, 1.20; 95% CI, 1.05-1.37). Age younger than 1 year, shorter transport time, and longer dispatch time were associated with increased risk of death. Being pulseless upon discovery and receiving CPR were associated with new neurologic dysfunction. Maximizing EMS transport interventions for patients younger than 1 year requiring CPI may improve patient outcomes.


Influence of mouse genetic background on wear particle-induced in vivo inflammatory osteolysis.
Genetics may influence wear particle-induced inflammatory osteolysis after joint replacement. In the present work, mice with three different genetic backgrounds were used to test this hypothesis. C57BL/6J, Balb/c and Kunming mouse were used. Each kind of mouse was divided into those receiving 30 mg UHMWPE particle implantation onto the calvariae and those receiving a sham operation. Mice of each group were sacrificed one week after surgery. Calvariae were harvested for immunological assay of TNF-alpha and IL-1 beta secretion in supernatants of calvariae organ culture and histological analysis of calvarial sagittal suture osteolysis and osteoclastogenesis. Although UHMWPE particles induced obvious calvarial sagittal suture osteolysis and osteoclastogenesis in all strains as compared with their corresponding control mice, the most significant change was found in C57BL/6J mice, less severe in Balb/c mice and much less severe in Kunming mice. In agreement with pathological findings, UHMWPE particles induced the highest IL-1 beta secretion in C57BL/6J mice, compared with Balb/c and Kunming mice. However, no difference was observed concerning TNF-alpha secretion among these mice. Our data suggests that genetics had a significant influence on wear particle-induced inflammation, osteoclastogenesis and osteolysis. The influence of genetic background on implant life in patients with joint replacement warrants further investigation.


Characterization and complete genome sequences of two novel variants of the family Closteroviridae from Chinese kiwifruit.
Two distinct closterovirus-like genome sequences (termed AdV-1 v1 and v2) were identified in Actinidia chinensis var. deliciosa 'Miliang-1' that had no disease symptoms using high-throughput sequencing. Using overlapping reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends, the genomic sequences of AdV-1 v1 and v2 were confirmed as 17,646 and 18,578 nucleotides in length, respectively. The two complete genomes contained 9 and 15 open reading frames, respectively, coding for proteins having domains typical of Closteroviridae, such as RNA-dependent RNA polymerase (RdRp), heat shock protein 70 homolog (HSP70h) and coat protein (CP). Sequence analysis showed that the amino acid sequences of RdRp, HSP70h, and CP of the two variants exhibited high similarity (> 80%), while their genomic organization was somewhat different. This suggested that the two viral genomes identified here are variants of the family Closteroviridae in a single kiwifruit host. Furthermore, phylogenetic relationship analysis revealed that the two variants had a closer relationship with the unclassified virus Persimmon virus B (PeVB) and Actinidia virus 1 (AcV-1) than with other members of the family Closteroviridae, as did their genomic organization. It is speculated that the two variants, together with PeVB and AcV-1 belong to a new subfamily of Closteroviridae.


Structural and chemical characterization of gallstones resistant to dissolution therapy.
X-ray diffraction, i.r. spectroscopic, and chemical analyses have been carried out on radiolucent gallstones resistant to dissolution therapy. Cholesterol represents the main component of all the examined stones, while the ratio between the amounts of pigmented material and calcium carbonate is about 1 in the inner and outer layers of the stones and 3 in the medial layer. Calcium carbonate is present in two distinct crystalline forms: vaterite, which is the main inorganic crystalline phase, and calcite. The cell parameters of vaterite and calcite are shorter in the inner and outer layers of the stones than in the medial layer. The observed variation of the cell parameters has been related to the substitution of copper to calcium in the carbonate structures, on the basis of the data obtained on vaterite and calcite synthesized in presence of different copper concentrations in solution. The results indicate that the failure of the dissolution therapy can be related to the inhomogeneous distribution in the stones of calcium carbonate and calcium bilirubinate.


Quantitation of putative colorectal cancer biomarker candidates in serum extracellular vesicles by targeted proteomics.
At the moment, there is no sensitive clinical test for detecting early-stage colorectal cancer (CRC). Target proteomics has enabled high-throughput verification of hundreds of biomarker candidate proteins. Using this technology, we verified 725 previously reported CRC biomarker candidate proteins that are functionally correlated with CRC in extracellular vesicles (EVs) from patients. Of these, 356 proteins were quantified, and 34 peptides (22 proteins) showed significant differences in the serum EVs between healthy controls and CRC patients of two independent cohorts (n = 77 and 84). These peptides were evaluated as single or multiple markers, and four single peptides in annexin family proteins and eight combinations of peptides showed area under the curve > 0.9 for discriminating between healthy controls and CRC patients. The sensitivities of annexins A3, A4, and A11 peptides for detecting early-stage CRC greatly exceed those of carcinoembryonic antigen. These peptides are promising biomarkers for early detection of CRC.


Construction of a thorium/actinium generator at the Canadian Nuclear Laboratories.
Targeted Alpha Therapy (TAT) has demonstrated considerable promise in the treatment of a range of cancers in both preclinical and, more recently clinical research. In particular, work with the alpha-emitting radionuclide 225Ac has been effectively employed due to the relatively rapid decay cascade that leads to 4 alpha and 2 beta emissions. One limitation for TAT has been caused by access to the vital radionuclide. Traditionally, 225Ac has been sourced from thorium/actinium generators based on the alpha decay of stockpiles of 229Th. 229Th is itself the alpha-decay product from 233U. Due to proliferation issues associated with 233U, only three thorium/actinium generators have been reported in the literature, capable of supporting clinical research. This paper describes the construction and operation of a thorium/actinium radionuclide generator at the Canadian Nuclear Laboratories, capable of supporting preclinical and limited clinical research in the area of TAT. Thorium was recovered and purified by a combination of anion exchange and extraction chromatography from aged 233U stockpiles. A separation scheme for 225Ra and 225Ac has been developed, based upon the chemical composition of the thorium material to allow for regular, routine milkings capable of supplying up to 3.7 GBq (100 mCi) of radiochemically pure 225Ac annually. This routine separation is accomplished using a combination of anion exchange chromatography to separate Ac and Ra isotopes from Th and extraction chromatography employing TEVA and DGA-N resins to separate actinium from radium and breakthrough thorium.


Fecal virome transfer improves proliferation of commensal gut Akkermansia muciniphila and unexpectedly enhances the fertility rate in laboratory mice.
Probiotics are intended to improve gastrointestinal health when consumed. However, the probiotics marketed today only colonize the densely populated gut to a limited extent. Bacteriophages comprise the majority of viruses in the human gut virome and there are strong indications that they play important roles in shaping the gut microbiome. Here, we investigate the use of fecal virome transplantation (FVT, sterile filtrated feces) as a mean to alter the gut microbiome composition to lead the way for persistent colonization of two types of probiotics: Lacticaseibacillus rhamnosus GG (LGG) representing a well-established probiotic and Akkermansia muciniphila (AKM) representing a putative next-generation probiotic. Male and female C57BL/6NTac mice were cohoused in pairs from 4 weeks of age and received the following treatment by oral gavage at week 5 and 6: AKM+FVT, LGG+FVT, probiotic sham (Pro-sham)+FVT, LGG+Saline, AKM+Saline, and control (Pro-sham+Saline). The FVT donor material originated from mice with high relative abundance of A. muciniphila. All animals were terminated at age 9 weeks. The FVT treatment did not increase the relative abundance of the administered LGG or AKM in the recipient mice. Instead FVT significantly (p < 0.05) increased the abundance of naturally occurring A. muciniphila compared to the control. This highlights the potential of propagating the existing commensal "probiotics" that have already permanently colonized the gut. Being co-housed male and female, a fraction of the female mice became pregnant. Unexpectedly, the FVT treated mice were found to have a significantly (p < 0.05) higher fertility rate independent of probiotic administration. These preliminary observations urge for follow-up studies investigating interactions between the gut microbiome and fertility.


Beneficial long-term effects of multiprofessional assessment vs. rehabilitation program in patients with musculoskeletal pain.
The aim of this study was to evaluate the differences in outcomes of long-term multiprofessional intervention in patients suffering from musculoskeletal pain. Eighty-eight patients with persistent musculoskeletal pain who either experienced multiprofessional rehabilitation (44) or underwent a mutiprofessional assessment of their ability to work (44) were followed up after either 5 or 10 years. Data on pain intensity, health perception and physical function collected in either 1998 or 2003, were compared with data taken from patients in 2008. Patients were also interviewed regarding pain diagnoses and additional multiprofessional interventions that occurred after 1998 or 2003. Results indicate that patients who participated in multiprofessional rehabilitation were younger in both 5- and 10-year follow-up groups and remained at work longer when compared to assessment group. When comparing the rehabilitation vs. assessment groups, no differences in pain intensity, health perception, and physical function were found at the 5- and 10-year follow-up. Reduced pain intensity was reported by both rehabilitation and assessment groups at the 5-year follow-up. Functional status was improved in the assessment group after 10 years. These results indicate that multiprofessional assessment may also have beneficial effects in patients with musculoskeletal pain when measuring long-term outcomes.


Thrombolytic therapy in ischemic stroke patients with pre-stroke depression in the telestroke vs non-telestroke.
Thrombolysis therapy remains the gold standard in acute ischemic stroke treatment, and rates of treatment with rtPA in ischemic stroke patients with comorbid depression has yet to be fully investigated. This study aims to examine clinical risk factors associated with inclusion or exclusion for rtPA in acute ischemic stroke populations with pre-stroke depression in the telestroke versus a non-telestroke setting. We collected retrospective data from a regional stroke registry for pre-stroke depressed ischemic stroke patients from January 2010 to June 2016. Logistic regression was used to determine demographic and baseline clinical risk factors associated with inclusion and exclusion from rtPA. . In the adjusted analysis, increasing age (OR = 1.064, 95% CI, 1.006-1.125, P = 0.029), improved ambulation (OR = 3.513, 95% CI, (0.855-14.436, P = 0.018) and sleep apnea (OR = 4.458, 95% CI, 0.731-27.182, P = 0.05) were associated with inclusion for rtPA, while Caucasian race (OR = 0.119, 95% CI, 0.0168-0.908, P = 0.040), systolic blood pressure (OR = 0.945, 95% CI, 0.906-0.985, P = 0.008), and direct admission (OR = 0.028, 95% CI, 0.003-0.317, P = 0.004) were associated with exclusion from rtPA. In the telestroke setting, INR (OR = 1.016, 95% CI, 0-5.393, P = 0.163) was not significantly associated with rtPA inclusion or exclusion. Identifying contraindicators associated with exclusion from rtPA is significant to improve the use thrombolytic therapy in the telestroke and non telestroke settings.


How getting twisted in scaffold design can promote bone regeneration: A fluid-structure interaction evaluation.
Bone tissue engineering (BTE) uses engineering principles to repair large bone defects, which requires effective mass transport ability of scaffolds to support cellular activities during bone regeneration. Since the implanted BTE scaffolds keep deforming under physiological loading which influences the fluid flow and mass transport within the scaffold and surrounding tissue, thus, scaffold design needs to consider the mass transport behavior under the physiological loading. This work proposed a novel twist scaffold, and its mass transport efficiency under physiological loading conditions was evaluated by a fluid-structure interaction analysis. The results showed that compared to the non-twist scaffold, the twist scaffold could form a rotating flow under the physiological loading, which enhanced the mass transport and generated more appropriate wall shear stress (WSS) to promote bone regeneration. This highlighted the better mass transport efficiency of the twist scaffold. Therefore, getting twist may be a promising design strategy for future BTE scaffolds, and the fluid-structure interaction approach may be a more reliable method for bone regeneration studies in either in vivo or in vitro systems.


"Dilute-and-inject" multi-target screening assay for highly polar doping agents using hydrophilic interaction liquid chromatography high resolution/high accuracy mass spectrometry for sports drug testing.
In the field of LC-MS, reversed phase liquid chromatography is the predominant method of choice for the separation of prohibited substances from various classes in sports drug testing. However, highly polar and charged compounds still represent a challenging task in liquid chromatography due to their difficult chromatographic behavior using reversed phase materials. A very promising approach for the separation of hydrophilic compounds is hydrophilic interaction liquid chromatography (HILIC). Despite its great potential and versatile advantages for the separation of highly polar compounds, HILIC is up to now not very common in doping analysis, although most manufacturers offer a variety of HILIC columns in their portfolio. In this study, a novel multi-target approach based on HILIC high resolution/high accuracy mass spectrometry is presented to screen for various polar stimulants, stimulant sulfo-conjugates, glycerol, AICAR, ethyl glucuronide, morphine-3-glucuronide, and myo-inositol trispyrophosphate after direct injection of diluted urine specimens. The usage of an effective online sample cleanup and a zwitterionic HILIC analytical column in combination with a new generation Hybrid Quadrupol-Orbitrap® mass spectrometer enabled the detection of highly polar analytes without any time-consuming hydrolysis or further purification steps, far below the required detection limits. The methodology was fully validated for qualitative and quantitative (AICAR, glycerol) purposes considering the parameters specificity; robustness (rRT < 2.0%); linearity (R > 0.99); intra- and inter-day precision at low, medium, and high concentration levels (CV < 20%); limit of detection (stimulants and stimulant sulfo-conjugates < 10 ng/mL; norfenefrine; octopamine < 30 ng/mL; AICAR < 10 ng/mL; glycerol 100 μg/mL; ETG < 100 ng/mL); accuracy (AICAR 103.8-105.5%, glycerol 85.1-98.3% at three concentration levels) and ion suppression/enhancement effects.


Angiopoietin-like protein 8 (betatrophin) inhibits hepatic gluconeogenesis through PI3K/Akt signaling pathway in diabetic mice.
Angiopoietin-like protein 8 (ANGPTL8) is a 198 amino-acid long, novel secreted protein that is mainly expressed in the liver and brown adipose tissues. At present, evidence supporting the involvement of ANGPTL8 in the regulation of glucose metabolism is inconclusive, along with its function in the liver. Previous studies mainly focused on the effect of ANGPTL8 on glucose metabolism in non-diabetic mice, and few relevant studies in diabetic mice exist. Therefore, this study aimed to investigate the role of ANGPTL8 on glucose homeostasis and elucidate the underlying mechanisms in diabetic mice. db/db diabetic and high-fat diet/streptozotocin-induced diabetic mice were injected with adenovirus expressing ANGPTL8 through the tail vein. Blood glucose levels were measured and glucose, insulin, and pyruvate tolerance tests were performed. To explore the molecular mechanism by which ANGPTL8 regulates hepatic glucose metabolism and manipulate mouse ANGPTL8 expression levels both in vivo and in vitro based on adenoviral transduction, gain- and loss-of-function strategies were adopted. Adenovirus-mediated overexpression of ANGPTL8 decreased fasting blood glucose levels and improved glucose tolerance and insulin sensitivity in db/db and high-fat diet/streptozotocin-induced diabetic mice. ANGPTL8 knockdown yielded the opposite effects. ANGPTL8 was upregulated in the cAMP/Dex-induced hepatocyte gluconeogenesis model. Moreover, ANGPTL8 overexpression in primary hepatocytes and diabetic mouse livers inhibited the expression of gluconeogenesis-related genes, including PEPCK and G6PC, by activating the AKT signaling pathway and, thereby, reducing glucose production. Therefore, the results demonstrated that ANGPTL8 improved glucose metabolism via inhibition of hepatic gluconeogenesis in diabetic mice. Current findings highlight a critical role of hepatic ANGPTL8 in glucose homeostasis, suggesting that increased ANGPTL8 expression could be an underlying factor for the inhibition of hepatic gluconeogenesis, which could be targeted for the prevention and treatment of type 2 diabetes.


Treatment results in patients with retinoblastoma and invasion to the cut end of the optic nerve.
There is little information on the outcome of patients with retinoblastoma and tumor at the resection margin of the optic nerve. Retrospective evaluation of three successive prospective protocols. Twenty-six consecutive patients were analyzed (International Staging System-IRSS-stage 2 = 21, stage 3 = 5) from three successive prospective protocols (1988-2006). Patients with stage 2 were enucleated upfront and those with stage 3 had neoadjuvant chemotherapy followed by enucleation and adjuvant therapy. Both groups received adjuvant chemotherapy and orbital radiotherapy after enucleation. Patients in protocol 1 received 1 year of the lower-dose chemotherapy regimen including cyclophosphamide, vincristine and doxorubicin along with intrathecal chemotherapy. Patients of protocols 2 and 3 received a more intense and shorter intravenous regimen including carboplatin and etoposide alternating with cyclophosphamide, idarubicin and vincristine with no intrathecal treatment. The components of protocol 2 and 3 were similar except for the dose of carboplatin which was 10% lower in protocol 3. Thirteen were treated in protocol 1 and 13 in protocols 2 and 3. The probability of event-free survival was 0.70 at 5 years. Events included: CNS relapse = 3, second malignancies = 3, death in complete remission = 2. There were no significant differences in outcome between protocols or stages. Endocrinological disturbances related to the hypothalamus-hypophysis axis were evident in 6/8 patients evaluated. Severe orbital sequelae occurred in 12 cases. A substantial number of patients with tumor at the resection margin of the optic nerve can be cured with current therapy; however, therapy related sequelae are frequent.


Induction and survival of micronuclei in rat spermatids. Comparison of two meiotic micronucleus techniques using cyclophosphamide.
The induction and survival of micronuclei (MN) in rat spermatids was studied by two different methods, the dissection method (DM) and the suspension method (SM). It was observed that MN are induced by cyclophosphamide in the S phase of meiosis, in preleptotene spermatocytes, and in an earlier cell stage, the type B spermatogonia. Both techniques showed that MN survive in spermatids at least 5 days. Advantages of the DM include the use of a DNA-specific fluorochrome for staining of MN, higher MN frequencies observed, and the possibility to gain detailed information of the kinetics of induction of MN. In the SM, slide preparation is simpler than in the DM, and several samples can be prepared simultaneously but the scoring of slides is time consuming. Improvements of the sampling system of the DM are suggested. For evaluation of clastogenic action of chemicals on male germ cells both techniques provide a simple and rapid approach.


Estimation of transpulmonary driving pressure during synchronized mechanical ventilation using a single lower assist maneuver (LAM) in rabbits: a comparison to measurements made with an esophageal balloon.
Mechanical ventilation is applied to unload the respiratory muscles, but knowledge about transpulmonary driving pressure (ΔPL) is important to minimize lung injury. We propose a method to estimate ΔPL during neurally synchronized assisted ventilation, with a simple intervention of lowering the assist for one breath ("lower assist maneuver", LAM). In 24 rabbits breathing spontaneously with imposed loads, titrations of increasing assist were performed, with two neurally synchronized modes: neurally adjusted ventilatory assist (NAVA) and neurally triggered pressure support (NPS). Two single LAM breaths (not sequentially, but independently) were performed at each level of assist by acutely setting the assist to zero cm H2O (NPS) or NAVA level 0 cm H2O/uV (NAVA) for one breath. NPS and NAVA titrations were followed by titrations in controlled-modes (volume control, VC and pressure control, PC), under neuro-muscular blockade. Breaths from the NAVA/NPS titrations were matched (for flow and volume) to VC or PC. Throughout all runs, we measured diaphragm electrical activity (Edi) and esophageal pressure (PES). We measured ΔPL during the spontaneous modes (PL_PES) and controlled mechanical ventilation (CMV) modes (PL_CMV) with the esophageal balloon. From the LAMs, we derived an estimation of ΔPL ("PL_LAM") using a correction factor (ratio of volume during the LAM and volume during assist) and compared it to measured ΔPL during passive (VC or PC) and spontaneous breathing (NAVA or NPS). A requirement for the LAM was similar Edi to the assisted breath. All animals successfully underwent titrations and LAMs for NPS/NAVA. One thousand seven-hundred ninety-two (1792) breaths were matched to passive ventilation titrations (matched Vt, r = 0.99). PL_LAM demonstrated strong correlation with PL_CMV (r = 0.83), and PL_PES (r = 0.77). Bland-Altman analysis revealed little difference between the predicted PL_LAM and measured PL_CMV (Bias = 0.49 cm H2O and 1.96SD = 3.09 cm H2O). For PL_PES, the bias was 2.2 cm H2O and 1.96SD was 3.4 cm H2O. Analysis of Edi and PES at peak Edi showed progressively increasing uncoupling with increasing assist. During synchronized mechanical ventilation, a LAM breath allows for estimations of transpulmonary driving pressure, without measuring PES, and follows a mathematical transfer function to describe respiratory muscle unloading during synchronized assist.


Automatic Methods for Carotid Contrast-Enhanced Ultrasound Imaging Quantification of Adventitial Vasa Vasorum.
Adventitial vasa vasorum are physiologic microvessels that nourish artery walls. In the presence of cardiovascular risk factors, these microvessels proliferate abnormally. Studies have reported that they are the first stage of atheromatous disease. Contrast-enhanced ultrasound (CEUS) of the carotid allows direct, quantitative and non-invasive visualization of the adventitial vasa vasorum. Hence, the development of computer-assisted methods that speed image analysis and eliminate user subjectivity is important. We developed methods for automatic analyses and quantification of vasa vasorum neovascularization in CEUS and tested these methods in a cohort of 186 individuals, 63 of whom were healthy volunteers. We implemented alternative automatic strategies for using the images to stratify patients according to their risk group and compare the strategies with respect to diagnostic performance. An automatic single-parameter strategy performs less effectively than the corresponding Arcidiacono method based on manual interpretation of the images (68 < area under the receiver operating characteristic curve [AUROC] for the manual Arcidiacono method < 82; 60 < AUROC for the automatic single-parameter strategy < 63). However, by use of additional image parameters, an automatic multiparameter strategy has significantly improved performance with respect to the manual Arcidiacono method (78 < AUROC < 90). The automatic multiparameter strategy is a valuable alternative to the manual Arcidiacono method, improving both diagnostic speed and diagnostic accuracy.


Hemodynamic changes following corticosteroid and naloxone infusion in dogs subjected to hypovolemic shock without resuscitation.
B-endorphin, which is released concomitantly with ACTH from the pituitary during stress, may also alter cardiac performance in hemorrhagic shock. In this study of 36 dogs subjected to hemorrhagic shock without resuscitation, we demonstrate the interaction of high doses of dexamethasone (DEX) or methylprednisolone (M) and opiate receptor blockade with naloxone (NAL). NAL, when given alone, resulted in the most hemodynamic improvement and the longest survival time while those animals receiving DEX or M, even in combination with NAL, did not do as well. These data suggest that corticosteroids block the NAL effect following hemorrhagic shock.


A conversation with Mariana Wolfner, newly inducted member of the National Academy of Sciences.
Molecular Reproduction and Development is delighted to announce that editorial board member Mariana F. Wolfner has been elected to the National Academy of Sciences. Here, Dr Wolfner is interviewed by two of her former postdocs. She discusses her path to studying reproduction and her career as a researcher and mentor.


Murine antiestrogen-binding protein: characterization, solubilization and modulation by lipids.
The properties of the antiestrogen-binding protein have been examined in mouse tissues, a species in which nonsteroidal antiestrogens are virtually pure agonists. As in other species studied, this protein was distributed in all tissues - highest levels being in the liver. Subcellular fractionation of mouse liver showed that 82% of the antiestrogen-binding protein was associated with the rough endoplasmic reticulum where it was confined to the membranous component. The antiestrogen-binding protein was also present in smooth endoplasmic reticulum, nuclei and cytosol. Its concentration in intact nuclei was at least 10-times higher than levels previously reported in intact rat liver nuclei. Binding of [3H]tamoxifen to the murine antiestrogen-binding protein was of high affinity (Kd = 1 nM) and was inhibited by unsaturated fatty acids and 7-ketocholesterol. In general, cis-isomers of unsaturated fatty acids were more effective binding inhibitors than trans-isomers. The antiestrogen-binding protein solubilized from rough endoplasmic reticulum membranes by the zwitterionic detergent CHAPS, had a molecular mass of approx. 700 kDa and a sedimentation coefficient of about 19 S. [3H]Tamoxifen binding capacity of the solubilized protein was abolished by trypsin and nonspecific proteinases but not by clostripain or Staphylococcus aureus V8 proteinase, suggesting that lysine residue(s) may be involved in [3H]tamoxifen binding.


Effects of systemic corticosteroid treatment on pseudotumoral hemicerebellitis: a case report and literature review.
Pseudotumoral hemicerebellitis is an acute, unilateral inflammation of the cerebellum that typically affects the pediatric population. The purpose of this paper is to review cases of pseudotumoral hemicerebellitis in the literature and evaluate if treatment with systemic corticosteroids reduces length of time to symptomatic recovery. We present a case report of a 12-year-old male with pseudotumoral hemicerebellitis and unilateral cerebellar dysfunction. Additionally, we review the thirty-five reported cases of pseudotumoral hemicerebellitis with respect to length of time to symptomatic recovery with or without systemic corticosteroid treatment. Thirty cases reported length of time to symptomatic recovery. Including our case, the mean time to recovery for those treated with systemic corticosteroids (n = 20) was 48.05 days (SE = 16.3). The mean time to recovery for those treated without (n = 10) was 86.7 days (SE = 29.3). Treatment with systemic corticosteroids was associated with a faster time to symptomatic recovery compared to without. Regardless of etiology, reducing inflammation and mass effect involved in pseudotumoral hemicerebellitis may be integral to a more rapid return to neurological baseline.


The number and size of neurons in the CNS of gastropod molluscs and their suitability for optical recording of activity.
Several factors that affect the suitability of opticaL methods for monitoring neuron activity were evaluated in several species of gastropod molluscs. The mean cell body diameter and the total number of cells in the central nervous system were determined In 6 preparations and qualitative evaluations were made for an additional 25 species. There was a factor of 10 difference in mean diameter between species with the smallest cells (prosobranchs) and those with the largest (certain opisthobranchs). Several opisthobranchs had about 5000 central neurons; we estimate that the prosobranchs and pulmonates had at least 5 times as many neurons. When the opacity to transmitted light was measured the percent transmissions of circumesophageal ganglia were between 4% and 40%. We attempted to measure optical signals associated with spike activity in 20 gastropod species; in most species signals were readily detected in single trials.


Study of the influence of temperature and precipitations on the levels of BTEX in natural waters.
Assessment of seasonal changes in surface water quality is an important aspect for evaluating temporal variation of water due to natural or anthropogenic inputs of point and non-point sources. The objective of this paper was to investigate the influence of seasonal temperature fluctuations and precipitations on the levels of BTEX in natural waters. Principal component analysis (PCA) was used to evaluate the seasonal correlations of BTEX levels in water and to extract the parameters that are most important in assessing seasonal variations of water quality. This study was carried out as a part of VOCs monitoring program in natural water samples from Mediterranean coast. To carry out this project, a multiresidue analytical method was used. The method was based on headspace solid-phase microextraction (HS-SPME) followed by gas chromatography coupled to flame ionization detector (FID). The limits of detection LODs found for the tested analyte tested were in the 0.001-1 μg/L range. These values were adequate for the analysis of these compounds in water samples according to the regulated values. Water samples from different points of the Mediterranean coast were analyzed during a period of three years, and were taken four times per year. Most of the compounds were below the limit established by the legislation. The results obtained by a chemometric study indicated that temperature and precipitations can be related on the BTEX levels found in water. A regression model between temperature or precipitations and BTEX concentration was obtained, thus these models can be used as predictive model for detection any non-normal concentration level.


Fluorescent Turn-On Sensing of Zinc(II) and Alkaline Phosphatase Activity Using a Pyridoxal-5'-Phosphate Derived Schiff Base.
A novel Zn2+ ion and alkaline phosphatase (ALP) selective fluorescence turn-on sensor L was developed by reacting pyridoxal 5'-phosphate (PLP) with hydrazine. Sensor L shows significant flurescence enhancement at 476 nm due to the formation of a L-Zn2+ complex in 1:1 binding stoichiometry with the association constant of 3.1⋅104 M- 1. Using L, the concentration of Zn2+ can be detected down to 2.34 µM, and the practical utility of L was validated by quantifying Zn2+ in real water samples. Additionally, the receptor L was applied to mimic the dephosphorylation reaction catalysed by the enzyme ALP and the resulted fluorescence change was monitored to detect the ALP activity.


Selection and structural analysis of the NY25 peptide - A vaccine candidate from hemagglutinin of swine-origin Influenza H1N1.
The aim of this study was to construct a vaccine peptide candidate against pandemic Influenza H1N1 hemagglutinin and to test its structure. With the help of bioinformatic algorithms we showed that the sequence encoding the second polypeptide of pandemic Influenza H1N1 hemagglutinin (HA2) is protected from nonsynonymous mutations better than the sequence encoding its first polypeptide (HA1). With the help of secondary and ternary structure predicting algorithms we found the fragment of HA2 with the most reproducible secondary structure and synthesized the NY25 peptide corresponding to the residues Asn117 - Tyr141 of HA2. According to the circular dichroism spectra analysis, the peptide has short helix and beta hairpin. According to the analysis of differential fluorescence quenching results, two tyrosine residues are situated on a long distance from each other. These facts taken together with the positive results of affine chromatography with the serum of a person immunized by full-length hemagglutinin confirm that the structure of the fragment of viral full-length protein has been reproduced in the synthetic NY25 peptide. Amino acid sequence of the NY25 peptide (NLYEKVRSQLKNNAKEIGNGCFEFY) is relatively conserved in 18 subtypes of Influenza A virus hemagglutinin.


Dietary omega-3 polyunsaturated fatty acid intake is related to a protective high-density lipoprotein subspecies profile independent of genetic effects: a monozygotic twin pair study.
Studies on diet and high-density lipoprotein (HDL) subspecies distribution are limited. We examined the relationship between macronutrient composition and lipoprotein particle size and HDL subspecies independent of genetic effects by studying monozygotic (MZ) twins. 24 healthy MZ twin pairs aged 23-33 years were identified from two longitudinal population-based studies, FinnTwin16 and FinnTwin12. Total energy and nutrient intake were assessed with 3-day food records and physical activity was measured by the Baecke index. HDL subspecies distribution was determined by non-denaturing gradient gel electrophoresis. Associations between diet composition and HDL mean particle size were determined by multivariate nutrient density models adjusted for confounding variables. Substituting one energy percentage from omega-3 polyunsaturated fatty acids (n-3 PUFAs) for a corresponding amount of energy from other type of fats was related to changes in the relative proportions of the HDL subspecies 2b, 3a and 3b toward a larger mean particle size in men (β ± SE: 1.00 ± 0.26 nm, p = 0.004) and women (β ± SE: 0.90 ± 0.21 nm, p = 0.001). This association remained significant in analyses controlling for genetic and shared environmental influences using within-pair differences of the measures in MZ twin pairs (β ± SE: 0.37 ± 0.14 nm, p = 0.019). Twins with the higher n-3 PUFA intake had significantly higher proportions of large HDL(2b) particles and lower proportions of smaller-sized HDL(3a) and HDL(3b) particles as compared to their co-twins with lower intakes (p < 0.05). Our data suggest that n-3 PUFA intake is associated with a favorable change in the distribution of HDL subspecies towards larger particles independent of genetic and shared environmental factors.


A simplified technique for revascularization of homografts of the liver with a variant right hepatic artery from the superior mesenteric artery.
A simplified technique for conversion of a complex hepatic arterial supply into a common channel is described. This technique permits single vessel anastomosis in the recipient of a liver transplant.


[Cutaneous adverse reactions to chemotherapy and their management].
Chemotherapy is an essential component in the multidisciplinary management of many cancers. Skin reactions to chemotherapeutics are rather frequent, and their morbidity may be significant. It is wise to identify them with precision in order to chose the most adequate preventive and therapeutic measures. The dermatopathology assessment is helpful for the diagnostic.


Control of hepatic gluconeogenesis: role of fatty acid oxidation.
Octanoate has been found to activate the gluconeogenic pathway in perfused isolated rat liver. Whether a net increase in the production of glucose is observed is a function of the relative concentrations of the glucose precursor and the fatty acid. The kinetics of octanoate interaction with the gluconeogenic pathway are influenced by the rate changes induced by decreases in pyruvate concentration as a result of the increased NAD redox potential produced by the oxidation of fatty acid. Taking this into account, two distinct effects of octanoate were identified. The first is an increase in the Vmax even at the lowest (25 microM) concentration of the fatty acid tested. The second is a progressive decrease in [pyruvate]0.5 as a function of octanoate concentration. The latter occurs at low (less than 0.1 mM), presumably physiological, pyruvate concentrations, when its mitochondrial transport is limiting, indicating that this process must have been activated. The former is observable even at high (greater than 0.5 mM), supraphysiological, concentrations of pyruvate, when its mitochondrial transport is not limiting, indicating that a distal step, presumably pyruvate carboxylation, is activated. The action of octanoate in increasing gluconeogenesis has been found not to be related to a decreased flux through pyruvate dehydrogenase, neither to changes in the NAD redox potential nor to its ability to increase energy production. Actually, the oxygen uptake induced by octanoate was largely accounted for by the production of ketone body and the latter process was found to be independent of variations in energy demand.


Protein chromatography on adsorbents with hydrophobic and ionic groups. Purification of human erythrocyte glycophorin.
Human erythrocyte glycophorin was purified rapidly by (a) chromatography of a Triton X-100 extract of erythrocyte 'ghosts' on N-(3-carboxypropionyl)aminodecyl-Sepharose in buffers containing Triton X-100 or sodium dodecyl sulphate, or (b) chromatography of whole 'ghosts', solubilized in sodium dodecyl sulphate, on dodecyl-Sepharose, in buffers containing sodium dodecyl sulphate. The products contained 85-95% glycophorin (electrophoretic band PAS-1) and the major contaminants were glycoproteins PAS-2 (possibly a subunit of glycophorin) and PAS-3.


Seasonality of root fungal colonization in low-alpine herbs.
Arbuscular mycorrhizal (AM) and dark septate endophytic (DSE) fungal colonization of Alchemilla glomerulans, Carex vaginata, Ranunculus acris ssp. pumilus and Trollius europaeus growing in low-alpine meadows in the Finnish subarctic were studied at different times during the growing season. Fungal colonization was correlated to soil soluble phosphorus (P) concentration. The influence of flower bud removal on fungal colonization was investigated in A. glomerulans, C. vaginata and R. acris and the correlation between AM and DSE colonization was studied. The fungal colonization patterns were found to be species-specific. R. acris maintained a relatively high rate of fungal colonization throughout the summer, while the rates of colonization of T. europaeus were lower and decreased towards the end of the season. A. glomerulans had constant arbuscular and vesicular colonization throughout the summer, but hyphal and DSE colonization declined towards the end of the season. C. vaginata did not form arbuscular mycorrhiza, but was colonized by DSE fungi and hyaline septate hyphae throughout the season. The soil soluble P concentration showed some seasonal variation, but was also highly variable between the study sites. Bud removal decreased arbuscular colonization of R. acris, but no unique effects were seen in any other parameters or the other species studied. The root fungal parameters correlated with soil P in some species at some sites, but no consistent trend was found. DSE colonization was positively correlated with root vesicular and hyphal colonization in some cases. The differences in fungal colonization parameters may be related to species-specific phenologies.


Dynamics of macromolecular populations: a mathematical model of the quantitative changes of RNA in the silkgland during the last larval instar.
The quantitative changes of RNA in the silkgland of Bombyx mori have been studied during the last larval instar by using a mathematical model (Volterra-Kostitzin model). This model can be associated with a global mechanism including synthesis and degradative processes. The numerical and statistical methods used for model analysis are described in an appendix. Thus we have compared the accumulation of total RNA (essentially ribosomal) after a treatment (juvenile hormone) and between several strains. The importance of the degradative factor is denoted to explain the observed differences, whereas the synthesis rates remain relatively stable. The last observation may lead us to an interpretation of the molecular effect of a selection to increase silk production : rather than an increase of the productivity of cellular machinery, the degradative process has been limited.


Treatment of Malignant Cutaneous Adnexal Neoplasms.
Malignant cutaneous adnexal neoplasms form a group of rare, typically low-grade-malignancy carcinomas with follicular, sebaceous, apocrine, or eccrine differentiation or a combination of the first 3 subtypes. Their clinical presentation is usually unremarkable, and biopsy is required to establish the differentiation subtype and the definitive diagnosis. Due to their rarity, no clear consensus has been reached on which treatment is most effective. Mohs micrographic surgery is considered to be the best option to prevent recurrence in the majority of patients. Radiotherapy and chemotherapy have been studied in very few cases and have rarely been shown to be effective.


The influence of lorazepam on somatosensory-evoked fast frequency (600 Hz) activity in MEG.
The generators of high frequency bursts (600-Hz activity) detected at the parietal scalp over the primary somatosensory cortex after electrical stimulation of peripheral nerves are not yet known. We investigated the influence of benzodiazepine on the somatosensory-evoked 600-Hz activity by means of neuromagnetic measurements and source analysis. After oral administration of lorazepam, the latency of the 600-Hz burst activity was increased; specifically later peaks were delayed more than earlier peaks. In contrast, the latency of the concurrent primary cortical low frequency response (N20m) was not significantly changed. The source strengths of both N20m and 600-Hz bursts were significantly increased. Our results provide evidence for two components of the 600-Hz activity with a different generator structure.


Early extracoronary atherosclerosis and coronary heart disease risk factors in a sample of civil servants in southern Italy.
One hundred and thirty-eight male subjects working as civil servants in the Catanzaro city hall, aged over 45, underwent Echo-Doppler examination of carotid and femoral arteries. Forty-one subjects (30%) presented evidence of early atherosclerotic lesions in at least 1 of the 10 examined arteries. Age, systolic blood pressure, serum triglycerides and serum glucose were higher in this group when compared with subjects without evidence of extracoronary atherosclerosis (EA), while HDL cholesterol and apolipoprotein A-I were lower. After adjustment for age, only serum triglycerides and apolipoprotein A-I levels remained significantly different. Only 21 subjects had no major coronary heart disease (CHD) risk factors, 46 had one, 47 two and 24 more than two risk factors. The prevalence of lesions was significantly higher in individuals with total cholesterol/HDL cholesterol ratio greater than 5. The presence of smoking and hypertension was almost equally represented in individuals with or without lesions. Serum triglycerides, total cholesterol/HDL cholesterol > 5 and apolipoprotein A-I seem to play a fundamental role in the development of EA, as detected by Duplex examination of carotid and femoral arteries. Age is also closely related to the presence of lesions, probably reflecting the exposure duration to CHD risk factors.


Traffic noise affects foraging behavior and echolocation in the Lesser Bulldog Bat, Noctilio albiventris (Chiroptera: Noctilionidae).
Urban noise, such as that of traffic, can affect the sensory capabilities in animals. Foragers acting optimally are expected to exploit feeding patches depending on the cost/benefit ratio, and some noises can cause increased foraging costs. We hypothesized that traffic noise affects foraging patch quality for aerial-insectivorous bats and predicted that there are measurable differences in their foraging activity and in their echolocation signals between nights with or without traffic noise. We tested these predictions in the Lesser Bulldog Bat, Noctilio albiventris, foraging over aquaculture fishponds, by recording bat activity on nights playing traffic noise and on traffic-sound free control nights. We measured foraging activity by counting the number of search passes and feeding buzzes made by the bats and measured several characteristics of echolocation signals. Foraging activity was higher during noisy nights than during control ones, probably due to the need of more time to obtain information about prey under noisy conditions. When exposed to traffic noise, the bats changed the spectral and temporal characteristics of echolocation signals. We suggest that exposure to traffic noise increases foraging costs for N. albiventris and posit that these bats continue foraging under noisy conditions because they can modulate their echolocation signals to obtain enough information.


Association of elevated glycosylated hemoglobin A1c with hyperfiltration in a middle-aged and elderly Chinese population with prediabetes or newly diagnosed diabetes: a cross-sectional study.
To examine whether elevated glycosylated hemoglobin A1c (HbA1c) is associated with hyperfiltration in a middle-aged and elderly Chinese population. Anthropometric and biochemical examinations were performed in 2491 individuals from the general population, aged 40-79 years, who participated in the Huaian Diabetes Prevention Program. The estimated glomerular filtration rate (eGFR) was calculated from creatinine levels using the CKD-EPI formula. Hyperfiltration was defined as eGFR >90(th) percentile. After adjustment [for age, gender, waistline, body mass index, blood pressure, smoking, alcohol consumption, cholesterol, log(triglycerides), high-density lipoprotein, low-density lipoprotein, serum uric acid, sodium intake, hypertension, and use of angiotensin converting enzyme inhibitors or angiotensin receptor blockers], HbA1c and fasting plasma glucose (FPG) were found to be independently positively associated with eGFR. Additionally, after multivariate adjustment, the odds ratios (95 % CI) for hyperfiltration calculated for a 1-unit increase in HbA1c and FPG were 1.396 (1.089-1.790) and 1.306 (1.117-1.526), respectively. Compared with participants with HbA1c levels <5.7%, the odds ratios (95 % CI) for hyperfiltration were 2.344 (1.025-5.364) in participants with HbA1c levels of 6.21-6.49%, and 2.965 (1.537-5.720) in those with HbA1c levels ≥ 6.5%. Elevated HbA1c (≥ 6.21%) is associated with an increased odds of hyperfiltration in middle-aged and elderly Chinese. Longitudinal studies are needed to explore whether hyperfiltration increases the odds of diabetic nephropathy in individuals with prediabetes.


[Association of GNB3 gene C825T polymorphism with coronary heart disease].
The C825T polymorphism in the gene encoding the G protein beta 3 subunit (GNB3) causes enhanced G protein activation and the increased in vitro cell proliferation. We investigated the association of gene GNB3 C825T polymorphism with coronary artery disease (CAD) in the Russian population. A total of 313 patients with CAD diagnosed on the basis of clinical studies and coronary angyography were examined. The control group included 132 individuals that lacked clinical CAD symptoms and had matching profile of coronary artery disease risk factors. Blood pressure was measured using standard protocols. Increased levels of diastolic and systolic pressure was observed in both groups. The allele and genotype frequencies of this polimorphic marker were significantly higher in the CAD patients than in control. We found that the frequency of allele C and gen-. otype CC was significantly higher in the CAD patients (OR = 1.55; P = 0.0079; OR = 1.63; P = 0.0215, respectively), which suggests higher risk of this pathology in carriers of allele C and genotype CC. Thus, in the Russian population coronary artery disease is associated with GNB3 allele C and genotype CC.


Creation of an extremely thin flap using IntraLase femtosecond laser.
A 43-year-old woman had laser in situ keratomileusis (LASIK) using the IntraLase femtosecond laser (IntraLASIK) to create 110 microm flaps. Despite uneventful flap formation, the flaps in both eyes were extremely thin, making it difficult to lift them. The flap in the right eye resembled an epi-LASIK epithelial flap. It was lifted uneventfully. The flap in the left eye was stretched and torn on lifting, and the procedure was therefore postponed. After 6 months, the IntraLASIK surgical procedure was repeated in the left eye using a thicker and smaller flap than in the first procedure. Despite the improvements associated with using an IntraLase femtosecond laser, thinner-than-intended corneal flaps can occur. Early recognition of such a flap can prevent further complications.


Reduced plasma coagulation factor XIII in patients with acute leukemia in remission during consolidation chemotherapy cycles.
Acute leukemia (AL) is a malignancy from hematologic stem cells (HSC). Consolidation with intensive chemotherapy is required after induced remission and repeatedly causes treatment-related bleeding that is usually attributed to chemotherapy-induced thrombocytopenia (CIT). However, our previous study demonstrated that severe deficiency of plasma coagulation factor XIII (pFXIII) also participated in the bleeding of CIT in AL. However, the relationship between pFXIII deficiency and consolidation chemotherapy was unknown. Here, we observed the concentration of pFXIII in patients with AL before and after consolidation chemotherapy and reevaluated the correlation to bleeding in myelosuppression. Thus, we found that the concentration of pFXIII before chemotherapy in all patients was markedly lower than in the control data and was further decreased by chemotherapy, related to bleeding in myelosuppression. These findings indicated that chemotherapy-induced pFXIII deficiency should be of concern and explored in depth.


Nitrogen allocation in Mojave Desert winter annuals.
Nitrogen contents and allocation were examined in winter annuals at two Mojave Desert sites near Boulder City, Nevada. Application of 10 g N m-2 as NH4NO3 increased production 0- to 7-fold in species growing on a sandy soil (an Entisol) but fertilizer had no effect on plants on an alluvium (an Aridisol). Tissue nitrogen comprised 0.09-3.5% of dry weight with the lowest concentrations found in vegetative organs of nitrogen-responsive plants. During development, nitrogenpoor species showed only minor changes in nitrogen concentration and allocation compared with more nitrogen-rich species. Maximum reproductive nitrogen allocation varied among species from 43 to 67%, while reproductive biomass allocation was 31 to 51%. Fertilizer increased reproductive biomass allocation by 7 to 16%, reproductive nitrogen concentrations by 120 to 260%, and eaf and root nitrogen concentrations by 200 to 615% in nitrogen-deficient plants. Nitrogen-poor plants appear to allocate nitrogen to reproduction at the expense of vegetative organs throughout the life cycle.


Overview of (1-->3)-beta-D-glucan immunobiology.
Glucans are (1-->3)-beta-D-glucose polymers that are found in the cell wall of fungi, bacteria and plants. Glucans are known to stim ulate humoral and cell-mediated immunity in humans and animals. In addition to the potent immune stimulatory effects of (1-->3)-beta-D-glucans, there are a number of toxicological effects associated with exposure to the water-insoluble, microparticulate form of the polymer. Recent investigations have suggested a potential role for (1-->3)-beta-D-glucans in inhalational toxicity. Specifically, (1-->3)-beta-D-glucans have been implicated in the symptomatology associated with 'sick building' syndrome. The mechanisms by which (1-->3)-beta-D-glucans mediate immune stimulation and, perhaps, toxicity are currently under investigation. It is now established that (1-->3)-beta-D-glucans are recognized by macrophages and, perhaps, neutrophils and natural killer cells via a (1-->3)-beta-glucan specific receptor. Following receptor binding, glucan modulates macrophage cytokine expression. Here we review the chemistry, immunobiology and toxicity of (1-->3)-beta-D-glucans and how it may relate to effects caused by inhalation.


Identification of differentially expressed genes in cucumber (Cucumis sativus L.) root under waterlogging stress by digital gene expression profile.
High-throughput tag-sequencing (Tag-seq) analysis based on the Solexa Genome Analyzer platform was applied to analyze the gene expression profiling of cucumber plant at 5 time points over a 24h period of waterlogging treatment. Approximately 5.8 million total clean sequence tags per library were obtained with 143013 distinct clean tag sequences. Approximately 23.69%-29.61% of the distinct clean tags were mapped unambiguously to the unigene database, and 53.78%-60.66% of the distinct clean tags were mapped to the cucumber genome database. Analysis of the differentially expressed genes revealed that most of the genes were down-regulated in the waterlogging stages, and the differentially expressed genes mainly linked to carbon metabolism, photosynthesis, reactive oxygen species generation/scavenging, and hormone synthesis/signaling. Finally, quantitative real-time polymerase chain reaction using nine genes independently verified the tag-mapped results. This present study reveals the comprehensive mechanisms of waterlogging-responsive transcription in cucumber.


Secretory leukocyte protease inhibitor binds to Neisseria gonorrhoeae outer membrane opacity protein and is bactericidal.
Secretory leukocyte protease inhibitor (SLPI) is an innate immune peptide present on the genitourinary tract mucosa that has antimicrobial activity. In this study, we investigated the interaction of SLPI with Neisseria gonorrhoeae. ELISA and far-Western blots were used to analyze binding of SLPI to gonococci. The binding site for SLPI was identified by tryptic digests and mass spectrometry. Antimicrobial activity of SLPI for gonococci was determined using bactericidal assays. SLPI protein levels in cell supernatants were measured by ELISA, and SLPI mRNA levels were assessed by quantitative RT-PCR. SLPI bound directly to the gonococcal Opa protein and was bactericidal. Epithelial cells from the reproductive tract constitutively expressed SLPI at different levels. Gonococcal infection of cells did not affect SLPI expression. We conclude that SLPI is bactericidal for gonococci and is expressed by reproductive tract epithelial cells and thus is likely to play a role in the pathogenesis of gonococcal infection.


Highlighting vulnerabilities in the alternative lengthening of telomeres pathway.
The alternative lengthening of telomeres (ALT) pathway is a telomere elongation mechanism found in a small but often aggressive subset of cancers. Dependent on break-induced replication, telomere extension in ALT-positive cells relies on a baseline level of DNA replication stress to initiate elongation events. This results in an elevated level of DNA damage and presents a possible vulnerability to be exploited in the development of ALT-targeted cancer therapies. Currently, there are no treatment options that target the ALT mechanism or that are specific for ALT-positive tumors. Here, we review recent developments and promising directions in the development of ALT-targeted therapeutics, many of which involve tipping the balance towards inhibition or exacerbation of ALT activity to selectively target these cells.


Carpometacarpal dislocation second to fifth with associated hamate fracture: A case report of rare injury.
Carpometacarpal (CMC) dislocation of the second to fifth metacarpals is an extremely rare hand injury that results from high-energy trauma. It is inherently unstable, especially if it is associated with concomitant carpal or metacarpal fractures. We report the case of an 18-year-old male who sustained a motorbike injury at the level of the second to fifth CMC joints, which was associated with a hamate fracture. The patient failed initial conservative treatment, and so one week later he underwent closed reduction of the second to fifth carpometacarpal dislocations and Kirschner wire fixations from the second to fifth metacarpals. He then underwent open reduction and internal fixation of the hamate fracture using a mini-fragment plate and screws. This type of hand trauma is often overlooked due to radiological misinterpretations. It is important to pay attention to significant soft tissue swelling in this injury. Percutaneous pinning is still considered the gold standard and safest procedure for this injury. This is a rare hand injury that requires careful radiological assessment and meticulous and stable reduction. We believe that open reduction is advantageous in certain circumstances.


In vitro activities of combinations of aztreonam, ciprofloxacin, and ceftazidime against clinical isolates of Pseudomonas aeruginosa and Pseudomonas cepacia from patients with cystic fibrosis.
The in vitro activities of two-drug combinations of aztreonam, ciprofloxacin, and ceftazidime were studied in 96 clinical isolates of Pseudomonas aeruginosa and in 20 clinical isolates of Pseudomonas cepacia from cystic fibrosis patients. Some synergy was observed with each combination used against P. aeruginosa, but synergy was rare when the combinations were used against P. cepacia.


Metabolic side effects of second generation antipsychotic agents in antipsychotic-naïve patients: one-month prospective evaluation.
The present study investigated the effects of second generation antipsychotics (SGA) on the metabolism of 15 antipsychotic-naïve outpatients. Evaluations were performed at baseline and after 1 month of treatment. A significant increase in mean body mass index (BMI) and mean waist circumference was observed. These results suggest the importance of monitoring patients from the first few weeks of antipsychotic treatment.


Spontaneous evolution of bicontinuous nanostructures in dealloyed Li-based systems.
Dealloying, the selective dissolution of one or more of the elemental components of an alloy, is an important corrosion mechanism and a technologically relevant process used to fabricate nanoporous metals for a variety of applications including catalysis, sensing, actuation, supercapacitors and radiation-damage-resistant materials. In noble-metal alloy systems for which the ambient-temperature solid-state diffusivity is minuscule, dealloying occurs at a composition-dependent critical potential above which bicontinuous nanoporous structures evolve and below which a full-coverage layer of the more-noble component forms causing the alloy surface to become passive. In contrast, for alloy systems exhibiting significant solid-state diffusive transport, our understanding of dealloying-induced morphologies and the electrochemical parameters controlling this are largely unexplored. Here, we examine dealloying of Li from Li-Sn alloys and show that depending on alloy composition, particle size and dealloying rate, all known dealloyed morphologies evolve including bicontinuous nanoporous structures and hollow core-shell particles. Furthermore, we elucidate the role of bulk diffusion in morphology evolution using chronopotentiometry and linear sweep voltammetry. Our results may have implications for lithium-ion battery development while significantly broadening the spectrum of strategies for obtaining new nanoporous materials through dealloying.


Patient satisfaction and symptom relief after anal dilatation.
Anal dilatation is used as a simple method of treatment and has been used for both anal fissure and haemorrhoids. This study examined longer-term results among a cohort of 162 patients, 132 of whom responded to a detailed questionnaire, an 82% response (66 patients were male; age range 17-75 years, median 42 years). Follow-up ranged from 16 months to 36 months (median 27 months) after anal dilatation (68 patients for fissure, 32 for haemorrhoids, and 32 for both). In the early months after dilatation, 83% had symptomatic improvement and 76% remained improved. Five (7%) patients with fissure and 11 with haemorrhoids (17%) required further hospital treatment, while 10% and 17%, respectively, had received further treatment from their general practitioners (GPs). Seventy-one percent said they would have a further anal dilation if symptoms recurred. There was no difference in results obtained by surgeons of different seniority. Complications--bleeding (29%) and difficulty controlling flatus (15%) or faeces (8%)--resolved in all cases. The results of anal dilatation for fissure are generally satisfactory in the longer term, with a trend toward better symptom relief in patients with fissure compared with those with haemorrhoids. We do not recommend anal dilatation as the sole treatment of patients with haemorrhoids, but it may be a useful adjunct to other treatments such as banding or sclerotherapy. Morbidity was generally acceptable and the majority of our patients would be prepared to have this procedure again if their symptoms were to return.


Communicating with Patients with Special Health Care Needs.
People with special health care needs (PSHCN) often have difficulty communicating with providers in health care settings, including dental practices. This difficulty can affect access to care as well as the quality of care received. This article provides practical tips and tools dental professionals can use to facilitate communication for a diverse population of PSHCNs. The article discusses communication needs of patients with communication disorders; augmentative and alternative communication; and communication for patients with intellectual disability, psychiatric conditions; and dental fears. Examples are given of communication breakdowns, and descriptions of how communication challenges can be resolved.


Imatinib modulates pro-inflammatory microenvironment with angiostatic effects in experimental lung carcinogenesis.
Lung cancer has second highest rate of incidence and mortality around the world. Smoking cigarettes is the main stream cause of lung carcinogenesis along with other factors such as spontaneous mutations, inactivation of tumor suppressor genes. The present study was aimed to identify the mechanistic role of Imatinib in the chemoprevention of experimental lung carcinogenesis in rat model. Gross morphological observations for tumor formation, histological examinations, RT-PCR, Western blotting, fluorescence spectroscopy and molecular docking studies were performed to elucidate the chemopreventive effects of Imatinib and support our hypothesis by various experiments. It is evident that immuno-compromised microenvironment inside solid tumors is responsible for tumor progression and drug resistance. Therefore, it is inevitable to modulate the pro-inflammatory signaling inside solid tumors to restrict neoangiogenesis. In the present study, we observed that Imatinib could downregulate the inflammatory signaling and also attributed angiostatic effects. Moreover, Imatinib also altered the biophysical properties of BAL cells such as plasma membrane potential, fluidity and microviscosity to restrict their infiltration and thereby accumulation to mount immuno-compromised environment inside the solid tumors during angiogenesis. Our molecular docking studies suggest that immunomodulatory and angiostatic properties of Imatinib could be either independent of each other or just a case of synergistic pleiotropy. Imatinib was observed to activate the intrinsic or mitochondrial pathway of apoptosis to achieve desired effects in cancer cell killings. Interestingly, binding of Imatinib inside the catalytic domain of PARP-1 also suggests that it has caspase-independent properties in promoting cancer cell deaths.


Discontinuous magnetic field effects (Barkhausen noise) in nucleic acids as evidence for room temperature organic superconduction.
Magnetization in fields of 2,000 to 9,000 gauss at room temperature generates low frequency noise in the microwave electrical conductivity of hydrated nucleic acids or dry melanin. The pattern of behavior observed for this noise suggests that it is analogous to Barkhausen noise observed during the penetration of bits of magnetic flux into type 2 superconductive metals. This implies that hydrated nucleic acids and dry melanin contain superconducting regions at room temperature.


The Relationship Between Social Risks and the Mental Health of School-Age Children in Primary Care.
The study's goal was to measure the association between social risks and the mental health of school-age children in primary care. We conducted a cross-sectional study in an urban safety-net hospital-based pediatric clinic using data collected from 2 standardized screening tools administered at well-child care visits for children age 6 to 11. Psychosocial dysfunction was measured with the Pediatric Symptom Checklist-17 (PSC-17), and 6 social risks (caregiver education, employment, child care, housing, food security, and household heat) were measured with the WE CARE screener. Multivariable linear and logistic regression analyses were conducted to measure the association between scores while controlling for sociodemographic characteristics. Among N = 943 patients, cumulative social risks were significantly associated with a positive PSC-17 total score (adjusted odds ratio [aOR] 1.2; 95% confidence interval [CI] 1.1-1.5; P = .02), indicating psychosocial dysfunction. Children with ≥3 social risks were 2.4 times more likely to have a positive PSC-17 total score compared to children with <3 social risks (95% CI 1.5-3.9; P < .001). Of the individual social risks measured, only food insecurity significantly predicted a positive PSC-17 total score (aOR 1.9; 95% CI 1.1-3.2; P = .02) and attention score (aOR 1.9; 95% CI 1.1-3.4; P = .03). Number of risks on a social risk screener was associated with psychosocial dysfunction in school-age children. Food insecurity was the only individual risk associated with psychosocial dysfunction, in particular attention problems. Screening tools for social risks could be used to identify at-risk children whose mental health may be adversely impacted by their social conditions.


The antigenic structure of the influenza virus A/PR/8/34 hemagglutinin (H1 subtype).
We have constructed an operational antigenic map of the hemagglutinin of influenza virus A/PR/8/34, which indicates the presence of five immunodominant antigenic regions exhibiting various degrees of operational linkage. These sites have been located by the identification of changed amino acid residues in mutant viruses that are antigenically altered at each site. Comparison of the antigenic features with the three-dimensional structure of the H3 subtype hemagglutinin shows that the antigenic sites correspond to four topographically distinct regions of the surface of the protein. One of the sites is formed when two regions that are widely separated in the hemagglutinin monomer associate in the assembled trimer. The location of the sites relative to those proposed for the H3 subtype hemagglutinin suggests that carbohydrate modulates the antigenicity of specific regions of the hemagglutinin.


The ubiquitin-like protein FAT10 in hepatocellular carcinoma cells limits the efficacy of anti-VEGF therapy by accelerating VEGF-independent angiogenesis.
The efficacy of anti-vascular endothelial growth factor (VEGF) therapy is limited. However, the key factors involved in limiting the efficacy of anti-VEGF therapy and the underlying mechanisms remain unclear. To investigate the effects and mechanisms of human leukocyte antigen F locus-adjacent transcript 10 (FAT10), a ubiquitin-like protein, in limiting the efficacy of anti-VEGF therapy in hepatocellular carcinoma (HCC) cells. FAT10 was knocked out in HCC cells using the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 technology. Bevacizumab (BV), an anti-VEGF monoclonal antibody, was used to evaluate the efficacy of anti-VEGF therapy in vivo. Mechanisms of FAT10 action were assessed by RNA sequencing, glutathione S-transferase pulldown assays and in vivo ubiquitination assays. FAT10 accelerated VEGF-independent angiogenesis in HCC cells which limited BV efficacy and BV-aggravated hypoxia and inflammation promoted FAT10 expression. FAT10 overexpression increased levels of proteins involved in several signaling pathways in HCC cells, resulting in upregulation of VEGF and multiple non-VEGF proangiogenic factors. Upregulation of multiple FAT10-mediated non-VEGF signals compensated for the inhibition of VEGF signaling by BV, enhancing VEGF-independent angiogenesis and promoting HCC growth. Our preclinical findings identify FAT10 in HCC cells as a key factor limiting the efficacy of anti-VEGF therapy and elucidate its underlying mechanisms. This study provides new mechanistic insights into the development of antiangiogenic therapies.


Procalcitonin (PCT) in patients with abdominal sepsis.
To assess the accuracy of procalcitonin as a measure of severity in patients with septic abdominal illnesses and the sepsis syndrome, to compare measurements with those of other inflammatory mediators, and to predict outcome. We carried out a prospective clinical study from 246 patients with infective or septic episodes confirmed at laparotomy and 66 patients undergoing elective operations who acted as controls. Specimens of blood for measurement of cytokine concentrations determination were obtained daily from septic patients. In the control group specimens were obtained before operation, at the end of operation, and on each of the following days until normal recovery (day 10). Every two weeks up to 3 months for patients with metastases, who were being followed up. Compared with other cytokines such as tumor necrosis factor alphaa and interleukin 6 procalcitonin was closely related to the development of infective and septic complications. 59 of 246 patients (24%) with sepsis died. Procalcitonin concentrations preoperatively [median 2.05 compared with 4.2 ng/ml (p=0.08)] (Mann-Whitney U-test) did not differ, but those on the days 1,4 and at the end differed significantly [day 1: 4.9 compared with 13.8 ng/ml (p<0.01); day 4: 4.8 compared with 13.0 ng/ml (p<0.01) and 0.4 compared with 13.25 ng/ml (p<0.01) at the end of the study]. In the control group only 7 (1.6%) of all blood samples, were detected outside the normal range (up to 0.8 ng/ml). Procalcitonin is a new indicator of infection and sepsis. TNF and IL-6 concentrations always rise after major operations and fall in the absence of infection, indicating operative trauma. Procalcitonin is sensitive in detecting infective complications. Under routine conditions the procalcitonin concentrations seems to be valid, reproducible and detectable.


[Working conditions of operating room personnel. Different symptoms observed and pregnancy outcome].
Neuropsychological symptoms and tiredness were reported more frequently by workers in less often scavenged theatres than by controls. The rate of spontaneous abortions was higher among pregnancies for which women were working in operating theatres.


Discovering unhealthiness: evidence from cluster analysis.
This study examines information on an array of health limitations, chronic conditions, treatments, and drug consumptions to reveal the prevalence and severity of unhealthiness that are not directly observed. Cluster analysis is applied to 265,468 individuals who participated in the 45 and Up Study in Australia. Among the study participants, 8% of those age 45-54 years, 10% of those age 55-64, 13% of those age 65-74, and 17% of those age 75 and older were classified as unhealthy. For the youngest individuals, unhealthiness is characterized by moderate-to-high mental distress, a poor physical health score equivalent to the score associated with having four major limitations in physical functioning, teeth health less than good, and having been diagnosed with at least two chronic conditions. The oldest individuals also suffer from these limitations, as well as dependence on at least three different drug groups and two medical treatments, but they are in better mental health state. Understanding unhealthiness across population groups will result in more effective allocation of health resources. Older populations require more resources to be devoted to the management of physical health and chronic illnesses.


Using Two Detection Methods to Observe the Changes and Significance of Free Light Chain in Serum and Urine in Patients with Renal Insufficiency.
Free light chains κ and λ (FLC κ, FLC λ) are of great significance in diagnostic and monitoring monoclonal gammopathy. Freelite and N-Latex methods are two common monitoring methods at present. But the two meanings are not completely equivalent, especially for patients with renal insufficiency. We analyzed the changes of serum and urine FLC in renal insufficiency patients without monoclonal gammopathy and the clinical significance of these changes. This study is an observational study. Patients ≥ 18 years old, who met the diagnostic criteria of chronic kidney disease (CKD), excluding monoclonal gammopathy, were selected. Fasting serum and 24-hour urine were taken to detect serum FLC κ, serum FLC λ, SCr, serum β 2-microglobulin, urinary FLC κ, urinary FLC λ, urinary α 1-microglobulin, and urinary β 2-microglobulin. There was a good correlation between the two methods for determining serum/urinary FLC. No matter serum or urine, FLC showed a good correlation with renal function by the N-Latex method, but not by the Freelite method. Under the N-Latex method, FLC κ/λ remained stable, which was basically within the reference range of healthy people and was not affected by renal function. There was a good correlation between FLC detected by N-Latex and microglobulin in serum and urine. When the concentration of FLC is low, the N-Latex method is more recommended to monitor FLC. The FLC measured by the N-Latex method is more closely related to renal function. The ratio of FLC κ/λ determined by the N-Latex method remained stable within the recommended range.


Examination of Large Artery Atherosclerosis could Reveal Small Artery Retinopathy in Untreated Middle-Aged Individuals.
Small arteries can be visualized in the ocular fundus, and findings of retinopathy based on Scheie classification are often applied to evaluate the impact of hypertension and atherosclerosis. However, the relationship between damage in the large and small arteries has not been investigated sufficiently, especially in the early stages. The present study investigated possible associations between large artery atherosclerosis and small artery retinopathy in untreated middle-aged individuals. Untreated middle-aged workers undergoing periodic health check-ups (n=7,730, 45±8 years) were enrolled in this study. The absence or presence and extent of retinopathy were characterized by ophthalmologists as hypertensive (H0-4) and atherosclerotic grades (S0-4) based on Scheie classification. Large artery atherosclerosis was examined based on functional assessment of the cardio-ankle vascular index (CAVI) and morphological assessment of the carotid intima-media thickness (IMT) by ultrasound. We found significant differences in CAVI and carotid IMT between individuals with and without hypertensive or atherosclerotic retinopathy. Multivariable regression analysis showed that the presence of hypertensive and atherosclerotic retinopathy was significantly associated with CAVI and carotid IMT. Logistic regression analysis with the endpoint of a hypertensive or atherosclerotic lesion revealed that CAVI and carotid IMT are independent determinants of retinopathy. CAVI and carotid IMT were significantly associated with the presence of retinopathy based on Scheie classification in untreated middle-aged subjects, implying that atherosclerotic examination in large arteries could reveal early-stage small artery retinopathy.


Autohydrolysis pretreatment assessment in ethanol production from agave bagasse.
The aim of the present work was to assess the autohydrolysis pretreatment of Agave tequilana bagasse for ethanol production. The pretreatment was conducted using a one-liter high pressure Parr reactor under different severity factors (SF) at a 1:6w/v ratio (solid:liquid) and 200rpm. The solids obtained under the selected autohydrolysis conditions were subjected to enzymatic hydrolysis with a commercial cellulase cocktail, and the enzymatic hydrolysate was fermented using Saccharomyces cerevisiae. The results obtained from the pretreatment process showed that the glucan content in the pretreated solid was mostly preserved, and an increase in the digestibility was observed for the case with a SF of 4.13 (190°C, 30min). Enzymatic hydrolysis of the pretreated solids showed a yield of 74.3%, with a glucose concentration of 126g/L, resulting in 65.26g/L of ethanol after 10h of fermentation, which represent a 98.4% conversion according to the theoretical ethanol yield value.


Determination of ofloxacin using a highly selective photo probe based on the enhancement of the luminescence intensity of Eu3+--ofloxacin complex in pharmaceutical and serum samples.
A rapid, simple and sensitive spectrofluorimetric method for determination of trace amount of ofloxacin was developed. At pH 5.1 the ofloxacin enhances the luminescence intensity of the Eu(3+) ion in Eu(3+)- ofloxacin complex at λ(ex) = 365 nm. The produced luminescence intensity of Eu(3+)-ofloxacin complex was in proportional to the concentration of ofloxacin. The working range for the determination of ofloxacin was 5.0 × 10(-9)-5.0 × 10(-6) mol L(-1) with lower detection limit (LOD) and quantitative detection limit (QDL) of 3 × 10(-9) and 9 × 10(-9) mol L(-1), respectively. The enhancement mechanism of the luminescence intensity in the Eu(3+)-ofloxacin system has been also explained. The method revealed good selectivity for ofloxacin in the presence of coexisting substances and used successfully for the assay of ofloxacin in pharmaceutical preparations and serum. A comparison with other standard methods was also discussed.


TAZ interacts with zonula occludens-1 and -2 proteins in a PDZ-1 dependent manner.
The transcriptional coactivator TAZ recognizes L/PPxY motifs in transcription factors like Runx1/2 through its WW domain. We show that the first PDZ domain of zona occludens-1 (ZO-1) and 2 (ZO-2) interacts with the carboxy-terminal PDZ binding motif of TAZ. Deletion of this motif abrogates binding. ZO-2 colocalizes with TAZ in the nucleus of MDCK cells and ZO-2 expression alters TAZ localization in human embryonic kidney cells. Luciferase assays demonstrate ZO-2 inhibition of TAZ-mediated transactivation. We propose that zonula occludens is a negative regulator of TAZ and suggest that selected tight junction proteins control nuclear translocation and activity of TAZ.