[Vascular changes in genesis of necrotic and inflammatory lesions of the lungs in Wegener's granulomatosis].
Clinico-morphological and histochemical studies of changes in the lungs were carried out in 15 fatal cases of Wegner's granulomatosis. Morphological manifestations of the lesions consisted in bilateral extensive destructive-productive panangiitis in the system of bronchial, was well as pulmonary arteries and veins. As a rule, there developed an extremely polymorphic granular tissue which subsequently underwent necrosis with resulting cavern formation. Deep and sometimes irreversible changes observed in the vessels of the microcirculatory bed led to alveolar-capillary blockade and hypoxia, the latter culminating with circulatory crisis. Drastic dyscirculatory disorders were complicated with auto- and superinfection: development of destructive-suppurative bronchitis, peribronchitis, interstitial and also confluent pneumonia with "motley" exudate in alveoli, not infrequently with an abscess formation, sometimes with formation of hyaline membranes. The whole complex of these changes play a significant part in pathogenesis of pulmonary-cardiac insufficiency occurring in the majority of patients with Wegener's granulomatosis and often is responsible for fatal outcomes.


Knowledge, adequacy, and approximate truth.
Approximation involves representing things in ways that might be close to the truth but are nevertheless false. Given the widespread reliance on approximations in science and everyday life, here we ask whether it is conceptually possible for false approximations to qualify as knowledge. According to the factivity account, it is impossible to know false approximations, because knowledge requires truth. According to the representational adequacy account, it is possible to know false approximations, if they are close enough to the truth for present purposes. In this paper, we adopt an experimental methodology to begin testing these two theories. When an agent provides a false and practically inadequate answer, both theories predict that people will deny knowledge. But the theories disagree about an agent who provides a false but practically adequate answer: the factivity hypothesis again predicts knowledge denial, whereas the representational adequacy hypothesis predicts knowledge attribution. Across two experiments, our principal finding was that people tended to attribute knowledge for false but practically adequate answers, which supports the representational adequacy account. We propose an interpretation of existing findings that preserves a conceptual link between knowledge and truth. According to this proposal, truth is not necessary for knowledge, but it is a feature of prototypical knowledge.


Diet modulates cecum bacterial diversity and physiological phenotypes across the BXD mouse genetic reference population.
The BXD family has become one of the preeminent genetic reference populations to understand the genetic and environmental control of phenotypic variation. Here we evaluate the responses to different levels of fat in the diet using both chow diet (CD, 13-18% fat) and a high-fat diet (HFD, 45-60% fat). We studied cohorts of BXD strains, both inbred parents C57BL/6J and DBA/2J (commonly known as B6 and D2, respectively), as well as B6D2 and D2B6 reciprocal F1 hybrids. The comparative impact of genetic and dietary factors was analyzed by profiling a range of phenotypes, most prominently their cecum bacterial composition. The parents of the BXDs and F1 hybrids express limited differences in terms of weight and body fat gain on CD. In contrast, the strain differences on HFD are substantial for percent body fat, with DBA/2J accumulating 12.5% more fat than C57BL/6J (P < 0.0001). The F1 hybrids born to DBA/2J dams (D2B6F1) have 10.6% more body fat (P < 0.001) than those born to C57BL/6J dams. Sequence analysis of the cecum microbiota reveals important differences in bacterial composition among BXD family members with a substantial shift in composition caused by HFD. Relative to CD, the HFD induces a decline in diversity at the phylum level with a substantial increase in Firmicutes (+13.8%) and a reduction in Actinobacteria (-7.9%). In the majority of BXD strains, the HFD also increases cecal sIgA (P < 0.0001)-an important component of the adaptive immunity response against microbial pathogens. Host genetics modulates variation in cecum bacterial composition at the genus level in CD, with significant quantitative trait loci (QTLs) for Oscillibacter mapped to Chr 3 (18.7-19.2 Mb, LRS = 21.4) and for Bifidobacterium mapped to Chr 6 (89.21-89.37 Mb, LRS = 19.4). Introduction of HFD served as an environmental suppressor of these QTLs due to a reduction in the contribution of both genera (P < 0.001). Relations among liver metabolites and cecum bacterial composition were predominant in CD cohort, but these correlations do not persist following the shift to HFD. Overall, these findings demonstrate the important impact of environmental/dietary manipulation on the relationships between host genetics, gastrointestinal bacterial composition, immunological parameters, and metabolites-knowledge that will help in the understanding of the causal sources of metabolic disorders.


Ultrastructural study of spermiogenesis and the spermatozoon of Microcotyle pancerii (Monogenea: Polyopisthocotylea: Microcotylidae), parasite of meagre Argyrosomus regius (Pisces: Teleostei).
The present work deals with the ultrastructure of spermiogenesis and the spermatozoon of Microcotyle pancerii, a gill parasite of meagre Argyrosomus regius collected in Corsican fish farms. Spermiogenesis was rather similar to that observed in other polyopisthocotylean Monogenea. The intercentriolar body was different from that described in digeneans. The nuclear condensation occurred in 2 successive stages. First, during the nuclear migration in the median cytoplasmic process, the nucleus developed a honeycomb-like appearance. Then, after the flagellar fusion, a discontinuous twisting of the chromatin appeared along the nucleus, with this process ending in total nuclear condensation. The structure of the spermatozoon is characterized by 2 axonemes (9 + "1" pattern), a single and continuous field of cortical microtubules, a mitochondrion, and a nucleus. Our findings were compared with various ultrastructural features in order to highlight variability within the group.


Effect of genome and environment on metabolic and inflammatory profiles.
Twin and family studies have established the contribution of genetic factors to variation in metabolic, hematologic and immunological parameters. The majority of these studies analyzed single or combined traits into pre-defined syndromes. In the present study, we explore an alternative multivariate approach in which a broad range of metabolic, hematologic, and immunological traits are analyzed simultaneously to determine the resemblance of monozygotic (MZ) twin pairs, twin-spouse pairs and unrelated, non-cohabiting individuals. A total of 517 participants from the Netherlands Twin Register, including 210 MZ twin pairs and 64 twin-spouse pairs, took part in the study. Data were collected on body composition, blood pressure, heart rate, and multiple biomarkers assessed in fasting blood samples, including lipid levels, glucose, insulin, liver enzymes, hematological measurements and cytokine levels. For all 51 measured traits, pair-wise Pearson correlations, correcting for family relatedness, were calculated across all the individuals in the cohort. Hierarchical clustering techniques were applied to group the measured traits into sub-clusters based on similarity. Sub-clusters were observed among metabolic traits and among inflammatory markers. We defined a phenotypic profile as the collection of all the traits measured for a given individual. Average within-pair similarity of phenotypic profiles was determined for the groups of MZ twin pairs, spouse pairs and pairs of unrelated individuals. The average similarity across the full phenotypic profile was higher for MZ twin pairs than for spouse pairs, and lowest for pairs of unrelated individuals. Cohabiting MZ twins were more similar in their phenotypic profile compared to MZ twins who no longer lived together. The correspondence in the phenotypic profile is therefore determined to a large degree by familial, mostly genetic, factors, while household factors contribute to a lesser degree to profile similarity.


[Effect of ginsenoside pre-treatment on 5-hydroxytryptamine system in SD rats with myocardial infarction and depression].
To evaluate the effect of ginsenoside pre-treatment on serotonin (5-hydroxytryptamine, 5-HT), 5-HT2A receptor (5-HT2AR), and 5-HT transporter (serotonin transporter, SERT) in serum, platelet lysate and brain tissue homogenates in SD rats with myocardial infarction (MI) or depression. Eighty SD rats were randomized treated with ginsenoside or normal saline(NS). After 4 weeks, the rats were, then, randomized to four subgroups: the MI, the depression, the MI in combination with depression and sham subgroups.5-HT, 5-HT2AR and SERT levels were detected in serum, platelet or brain. In the NS pre-treatment groups, serum 5-HT levels in sham rats (237.1±32.0) pg/ml were higher than in MI (58.0±11.6) pg/ml, depression (72.8±2.3) pg/ml, and MI with depression rats (62.5±10.2) pg/ml (all P=0.000). Pre-treatment of ginsenoside increased serum and platelet 5-HT in MI, depression and MI with depression rats compared with each corresponding NS treated subgroups (all P<0.005). However, treatment of ginsenoside lowered the 5-HT2AR expression in MI, depression, and MI with depression subgroups both in platelet and brain tissue homogenates compared with each corresponding NS treated subgroups (all P<0.05). As to SERT in platelet, treatment of ginsenoside increased its levels in depression rats (P=0.019), but lowered the levels in MI rats (P=0.001) compared with NS treated rats.There were no differences in brain 5-HT and SERT between ginsenoside and NS treated groups. The 5-HT system responses differently to ginsenoside treatment under different cardiac and mental stress conditions.Ginsenoside plays an important role in regulation of 5-HT system.


High-fidelity DNA polymerase enhances the sensitivity of a peptide nucleic acid clamp PCR assay for K-ras mutations.
Sensitive detection of tumor-specific point mutations is of interest in both the early detection of cancer and the monitoring of treatment at a molecular level. Recently, peptide nucleic acid (PNA) clamp real-time PCR has provided a time-sparing and sensitive method for the detection of mutations in the presence of a large excess of wild-type DNA. We present the first report that the sensitivity of PNA clamp PCR is limited by the low fidelity of TaqDNA polymerase. Replication errors introduced by Taq polymerase in the PNA-binding site were amplified during PCR due to the resulting mismatches between PNA and DNA. To reduce the frequency of polymerase-induced errors, we developed a PNA clamp PCR assay for the detection of mutations in codons 12 and 13 of the K-ras gene based on a high-fidelity DNA polymerase. The sensitivity of our assay increased approximately 10-fold, significantly detecting mutant DNA diluted 20,000-fold in wild-type DNA (P = 0.025), compared with its detection at 2000-fold dilution (P = 0.039) when Taq polymerase was used. Our data suggest that the replication errors caused by Taq polymerase must be taken into consideration for PNA clamp PCR and for other methods based on selective PCR amplification, and that these assays can be enhanced by high-fidelity DNA polymerases.


Differences in Growth Rate on CT of Adrenal Adenomas and Malignant Adrenal Nodules.
OBJECTIVE. The purpose of this study is to determine the differences in growth rate of adrenal adenomas and malignant adrenal nodules. MATERIALS AND METHODS. This was a retrospective review of adults with an adrenal nodule seen at two different abdominal or chest CT examinations or PET/CT examinations. Patients in the adenoma group were included if they had a CT, MRI, or pathologic diagnosis of an adrenal adenoma. Patients in the malignant group were included if they had a pathologically proven malignant adrenal nodule. Nodule growth was defined as a change in the largest axial diameter greater than or equal to 3 mm. Growth rate was calculated by dividing the change in the longest axial diameter by the time between the first and last imaging examination. RESULTS. There were 105 adenomas and 26 malignant nodules. Of the 105 adenomas, 34 (32.4%; 95% CI, 23.6-42.2%) grew, three (2.9%; 95% CI, 0.6-8.1%) became smaller, and 68 (64.8%; 95% CI, 54.8-73.8%), were unchanged in size. All 26 (100%; 95% CI, 89.1-100%) malignant nodules grew. The mean (± SD) growth rate of adenomas was 1.0 ± 0.67 mm/year (range, 0.3-2.8 mm/year), compared with 58.4 ± 78.5 mm/year (range, 5.8-395.4 mm/year) for malignant nodules (p < 0.001). A growth rate of 3 mm/year distinguished adenomas from malignant nodules with a sensitivity of 100% (95% CI, 86.8-100%) and a specificity of 100% (95% CI, 96.6-100%). CONCLUSION. Approximately one-third of radiologically proven adrenal adenomas grew, all of which grew at a rate less than 3 mm/year. All malignant adrenal nodules grew, and all at a rate greater than 5 mm/year.


Identification of adenovirus, influenza virus, parainfluenza virus, and respiratory syncytial virus by two kinds of multiplex polymerase chain reaction (PCR) and a shell vial culture in pediatric patients with viral pneumonia.
Early identification of causative agents in lower respiratory infection of pediatric patients can reduce morbidity and prevent an overuse of antimicrobials. Two kinds of multiplex polymerase chain reaction (PCR) and a commercial shell vial viral culture were performed to identify causative agents in pediatric patients. Nasopharyngeal aspirates of 220 children diagnosed with viral pneumonia were obtained. Two kinds of multiplex PCR (Seeplex(TM) RV detection kit, and Labopass(TM) RV detection kit), and a shell vial culture by R-Mix were performed. Positive samples from 220 total samples by two multiplex PCRs were 52.7% and 46.4%, respectively. We also cultured 103 samples that showed positive results of the adenovirus, influenza virus, parainfluenza virus, and respiratory syncytial virus (RSV) by two multiplex PCR. The RSV was most frequently detected in 53.0% (Seeplex) and 51.7% (Labopass) of patients. The detection rate of adenovirus (AdV) was 10.3% and 12.1%, influenza virus (IFV) A and B was 12.5% and 3.4%, and parainfluenza virus (PIFV) 1, 2, and 3 were 2.9% and 2.6%. Shell vial cultures showed concordant results with each multiplex PCR by 96.1% and 77.7%, respectively. Sequencing results were 90% consistent with multiplex PCR. Multiplex PCR showed more positivity than the shell vial culture and it can be an effective primary test. Other complementary efforts such as viral cultures and sequencing analysis could be considered, according to clinical and laboratory conditions.


Comparison of initial high-resolution computed tomography (HRCT) features of coronavirus disease 2019 (COVID-19) pneumonia and other viral pneumonias.
Multicenter retrospective comparison of the first high-resolution computed tomography (HRCT) findings of coronavirus disease 2019 (COVID-19) and other viral pneumonias. We retrospectively collected clinical and imaging data from 262 cases of confirmed viral pneumonia in 20 hospitals in Yunnan Province, China, from March 1, 2015 to March 15, 2020. According to the virus responsible for the pneumonia, the pneumonias were divided into non-COVID-19 (141 cases) and COVID-19 (121 cases). The non-COVID-19 pneumonias comprised cytomegalovirus (CMV) (31 cases), influenza A virus (82 cases), and influenza B virus (20 cases). The differences in the basic clinical characteristics, lesion distribution, location and imaging signs among the four viral pneumonias were analyzed and compared. Fever and cough were the most common clinical symptoms of the four viral pneumonias. Compared with the COVID-19 patients, the non-COVID-19 patients had higher proportions of fatigue, sore throat, expectorant and chest tightness (all P<0.000). In addition, in the CMV pneumonia patients, the proportions of acquired immunodeficiency syndrome (AIDS) and leukopenia were high (all PP<0.000). Comparison of the imaging findings of the four viral pneumonias showed that the pulmonary lesions of COVID-19 were more likely to occur in the peripheral and lower lobes of both lungs, whereas those of CMV pneumonia were diffusely distributed. Compared with the non-COVID-19 pneumonias, COVID-19 pneumonia was more likely to present as ground-glass opacity, intralobular interstitial thickening, vascular thickening and halo sign (all PP<0.05). In addition, in the early stage of COVID-19, extensive consolidation, fibrous stripes, subpleural lines, crazy-paving pattern, tree-in-bud, mediastinal lymphadenectasis, pleural thickening and pleural effusion were rare (all PP<0.05). The HRCT findings of COVID-19 pneumonia and other viral pneumonias overlapped significantly, but many important differential imaging features could still be observed.


MidA is a putative methyltransferase that is required for mitochondrial complex I function.
Dictyostelium and human MidA are homologous proteins that belong to a family of proteins of unknown function called DUF185. Using yeast two-hybrid screening and pull-down experiments, we showed that both proteins interact with the mitochondrial complex I subunit NDUFS2. Consistent with this, Dictyostelium cells lacking MidA showed a specific defect in complex I activity, and knockdown of human MidA in HEK293T cells resulted in reduced levels of assembled complex I. These results indicate a role for MidA in complex I assembly or stability. A structural bioinformatics analysis suggested the presence of a methyltransferase domain; this was further supported by site-directed mutagenesis of specific residues from the putative catalytic site. Interestingly, this complex I deficiency in a Dictyostelium midA(-) mutant causes a complex phenotypic outcome, which includes phototaxis and thermotaxis defects. We found that these aspects of the phenotype are mediated by a chronic activation of AMPK, revealing a possible role of AMPK signaling in complex I cytopathology.


Tumour necrosis factor-alpha polymorphisms and hepatocellular carcinoma: a meta-analysis.
The genetic basis of susceptibility to hepatocellular carcinoma (HCC) is poorly understood. To summarize the quantitative association between polymorphisms of the tumour necrosis factor-alpha (TNFA) gene and HCC, a meta-analysis of relevant studies was performed. Ten case-control studies involving 1421 HCC cases were identified from the Medline, Embase and Current Contents databases. Combined results based on all studies showed that patients with HCC had a significantly lower frequency of the TNFA gene polymorphism -308GG than healthy controls. When stratifying for race, results were similar among Asians and Caucasians. When comparing with hepatitis B virus infection cases, no statistical association was found. This meta-analysis suggests that TNFA -308GG gene polymorphism is associated with a modest decrease in the risk of HCC.


The serum inhibin B concentration and reference ranges in normozoospermia.
Although an inhibin B assay may be useful in the assessment of testicular function in a number of genital conditions, reliable reference ranges are still lacking. The present study sought to establish the reference range for serum inhibin B by applying the updated Gen II assay. This prospective study included 818 men referred for semen analysis: 377 were normozoospermic (reference group) and 441 presented at least one abnormal semen parameter (case group). Semen parameters were interpreted according to the 2010 World Health Organization manual and David's modified classification for normal morphology. The inhibin B concentration was determined with the current ELISA. In the reference group, the 2.5th percentile for inhibin B was 92 pg/ml and the 97.5th percentile for FSH was 7.8 IU/l. In the overall population, an inhibin B level <92 pg/ml was associated with increased odds ratio (OR; 95% CI) for oligozoospermia (16.93 (9.82-29.18), P<0.0001), asthenozoospermia (4.87 (2.88-8.10), P<0.0001), and teratozoospermia (2.20 (1.31-3.68), P=0.0026). The combination of a FSH >7.8 IU/l and an inhibin B <92 pg/ml was associated with greater OR for oligozoospermia (98.74 (23.99-406.35), P<0.0001) than for each hormone considered separately. A new reference range for serum inhibin B was established by the use of updated immunoassay. The correlations between hormone levels and semen parameters highlighted the importance of establishing these values with respect to the spermogram. When combined with FSH assay, the inhibin B range may be of value in the evaluation of spermatogenesis in a number of male genital conditions.


Usefulness of barium sulfate and iohexol as contrast agents for VFSS in visualizing components of swallowing predictable of poor outcomes.
Barium sulfate and iohexol are commonly used as contrast agents for videofluoroscopic swallowing study (VFSS). This study compared their usefulness as contrast agents in visualizing components of swallowing predictable of subsequent pneumonia and unintentional weight loss after VFSS. This was a randomized, controlled, crossover trial. The two contrast agents were alternately used in the same participants, and the order in which the contrast agent was tested first was randomly assigned. After VFSS, we followed the participants for 3 months and the association between VFSS findings of each contrast agent and the subsequent pneumonia and unintentional weight loss were analyzed. A total of 30 participants were included in the analysis. We recorded 11 cases of subsequent pneumonia and 13 of unintentional weight loss. Regarding the risk of subsequent pneumonia after VFSS, only the oral transit time and number of swallows tested with barium sulfate indicated significant differences between participants with and without subsequent pneumonia. For unintentional weight loss, oral transit time and pharyngeal wall coating after swallowing tested with barium sulfate, as well as oral transit time, nasal penetration, residue in the valleculae, PAS scores, and number of swallows when testing with iohexol demonstrated significant differences between those with and without unintentional weight loss.


Chromosomal genome assembly of the ethanol production strain CBS 11270 indicates a highly dynamic genome structure in the yeast species Brettanomyces bruxellensis.
Here, we present the genome of the industrial ethanol production strain Brettanomyces bruxellensis CBS 11270. The nuclear genome was found to be diploid, containing four chromosomes with sizes of ranging from 2.2 to 4.0 Mbp. A 75 Kbp mitochondrial genome was also identified. Comparing the homologous chromosomes, we detected that 0.32% of nucleotides were polymorphic, i.e. formed single nucleotide polymorphisms (SNPs), 40.6% of them were found in coding regions (i.e. 0.13% of all nucleotides formed SNPs and were in coding regions). In addition, 8,538 indels were found. The total number of protein coding genes was 4897, of them, 4,284 were annotated on chromosomes; and the mitochondrial genome contained 18 protein coding genes. Additionally, 595 genes, which were annotated, were on contigs not associated with chromosomes. A number of genes was duplicated, most of them as tandem repeats, including a six-gene cluster located on chromosome 3. There were also examples of interchromosomal gene duplications, including a duplication of a six-gene cluster, which was found on both chromosomes 1 and 4. Gene copy number analysis suggested loss of heterozygosity for 372 genes. This may reflect adaptation to relatively harsh but constant conditions of continuous fermentation. Analysis of gene topology showed that most of these losses occurred in clusters of more than one gene, the largest cluster comprising 33 genes. Comparative analysis against the wine isolate CBS 2499 revealed 88,534 SNPs and 8,133 indels. Moreover, when the scaffolds of the CBS 2499 genome assembly were aligned against the chromosomes of CBS 11270, many of them aligned completely, some have chunks aligned to different chromosomes, and some were in fact rearranged. Our findings indicate a highly dynamic genome within the species B. bruxellensis and a tendency towards reduction of gene number in long-term continuous cultivation.


Chitosan-based particulate systems for drug and vaccine delivery in the treatment and prevention of neglected tropical diseases.
Neglected tropical diseases (NTDs) are a diverse group of infections which are difficult to prevent or control, affecting impoverished communities that are unique to tropical or subtropical regions. In spite of the low number of drugs that are currently used for the treatment of these diseases, progress on new drug discovery and development for NTDs is still very limited. Therefore, strategies on the development of new delivery systems for current drugs have been the main focus of formulators to provide improved efficacy and safety. In recent years, particulate delivery systems at micro- and nanosize, including polymeric micro- and nanoparticles, liposomes, solid lipid nanoparticles, metallic nanoparticles, and nanoemulsions, have been widely investigated in the treatment and control of NTDs. Among these polymers used for the preparation of such systems is chitosan, which is a marine biopolymer obtained from the shells of crustaceans. Chitosan has been investigated as a delivery system due to the versatility of its physicochemical properties as well as bioadhesive and penetration-enhancing properties. Furthermore, chitosan can be also used to improve treatment due to its bioactive properties such as antimicrobial, tissue regeneration, etc. In this review, after giving a brief introduction to neglected diseases and particulate systems developed for the treatment and control of NTDs, the chitosan-based systems will be described in more detail and the recent studies on these systems will be reviewed. Graphical abstract.


A blinded pilot study of artwork in a comprehensive epilepsy center population.
The production of artwork is a complex neurological task. A controlled study of artwork produced by people with epilepsy has not previously been performed. The present report details the results of a three-part study involving 60 subjects from a comprehensive epilepsy center population. Subjects were grouped by the following diagnoses: seizures, partial seizures, complex partial seizures with temporal focus, and nonepileptic events. Data were collected in a blinded fashion. The Formal Elements Art Therapy Scale task showed significant effects in patients with epileptic seizures. The Free Drawing was most sensitive to complex partial seizures with temporal focus, while the Outline was most predictive of nonepileptic events. In addition to giving some insight into the neurological functioning of these subjects, this pilot study provides a basis for the future development of diagnostic tests to be used within this patient group.


Theory of infrared microspectroscopy for intact fibers.
Infrared microspectroscopy is widely used for the chemical analysis of small samples. In particular, spectral properties of small cylindrical samples are important in forensic analysis, understanding relationships between microstructure and mechanical properties in fibers or fiber composites, and development of cosmetics and drugs for hair. The diameters of the constituent cylinders are typically of the order of the central wavelength of light used to probe the sample. Hence, structure and material spectral response are coupled and recorded spectra are usually distorted to the extent of becoming useless for molecular identification. In this paper, we apply rigorous optical theory to predict the spectral distortions observed in IR microspectroscopic data of fibers. The theory is used, first, to compute the changes that are observed for cylinders of various dimensions under different instrument configurations when compared to the bulk spectrum from the same material. We provide a method to recover intrinsic material spectral response from fibers by correcting for distortion introduced by the cylindrical structure. The theory reported here should enable the routine use of IR microspectroscopy and imaging for the molecular analysis of cylindrical domains in complex materials.


Videofluoroscopic measures of swallowing in people with stable COPD compared to healthy aging.
Swallowing impairment is a serious extra-pulmonary manifestation of Chronic Obstructive Pulmonary Disease (COPD). Previous studies suggest that individuals with stable COPD show atypical values for several videofluoroscopy measures of swallowing, compared to healthy adults under age 60. However, it is unclear to what degree these changes are attributable to healthy aging. In this study, we aimed to clarify how swallowing in people with stable COPD differs from age-matched healthy controls. We performed a retrospective analysis of videofluoroscopy data from two previously-collected datasets: a) a sample of 28 adults with stable COPD (18 male); b) a sample of 76 healthy adults, from which 28 adults were selected, matched for sex and age to participants in the COPD cohort. In both prior studies, participants swallowed 20% w/v liquid barium prepared in different consistencies (thin; mildly, moderately, and extremely thick). Blinded duplicate ratings were performed according to a standard procedure, yielding measures of laryngeal vestibule closure (LVC) integrity and timing, swallow timing, upper esophageal sphincter (UES) opening, pharyngeal constriction and pharyngeal residue. Mann-Whitney U tests and odds ratios were performed to determine significant group differences (p<.05). Across the consistencies tested, participants with COPD showed significantly shorter durations of LVC, earlier onsets and shorter durations of UES opening, and reduced pharyngeal constriction. No significant differences were seen in other measures. These results point to features of swallowing in people with stable COPD that differ from changes seen with healthy aging, and which represent risks for potential aspiration.


The development of antibiotic resistant organisms with the use of ototopical medications.
There is growing concern over the use of systemic antibiotics and the development of bacterial resistance. The question remains as to whether ototopical medications may also promote antibiotic-resistant organisms, either on a local level (in the ear) or in other areas of the aerodigestive tract. We performed an evidence-based review to answer the following clinical question, "Do antibiotic ototopical medications induce antibiotic resistant organisms?" We performed a MEDLINE search of the published literature from 1966 to the present. We used appropriate search terms such as "ototopical antibiotics," "ototopical drops," "antibiotic resistance," "topical antibiotics and otitis externa," "otitis externa and treatment," "otitis externa and antibiotic drops," "otitis externa and ototopical drops," "otitis media," "otitis media and treatment," "otitis media and antibiotic drops," "chronic suppurative otitis media," "chronic suppurative otitis media and treatment," "chronic suppurative otitis media and antibiotic drops," " otitis externa and resistant organisms," "otitis media and resistant organisms," "chronic suppurative otitis media and resistant organisms," "ophthalmic antibiotic drops," "draining ear," "P.E. tube otorrhea," "pressure equalizing tube otorrhea," "pressure equalizing tube otorrhea and treatment," and "pressure equalizing tube otorrhea and ototopical therapy" to identify pertinent articles. These articles were reviewed and graded according to the evidence quality. After an initial screening of over 2,500 articles, 38 articles were analyzed further; of these, 11 were determined to warrant extensive review. Eight articles evaluated chronic suppurative otitis media; 2, otitis externa; and 1, post-tympanostomy tube otorrhea, whereas 3 others studied systemic absorption. Of the 8 chronic suppurative otitis media studies, there were thought to be 5 grade 2B studies, 1 grade 1B study, and 1 grade 2C study. These studies did not demonstrate a propensity for the development of resistant organisms. No study answered the question as to whether resistance to systemic antibiotics might occur in otitis externa. Overall grade B evidence seems to indicate that no significant antibiotic resistance develops from the use of ototopical antibiotic treatment.


Relationship between DNA damage and micronucleus in mouse liver.
To determine the optimum timing of partial hepatectomy (PH) in a previously developed mouse liver micronucleus test (Igarashi and Shimada, 1997), the relation between DNA damage and micronucleus was examined using the in vivo alkaline comet assay and the micronucleus test on the liver of the same individual mouse. Five genotoxic carcinogens, 1-nitropyrene (1-NP) (125 mg/kg), cyclophosphamide (CP) (50 mg/kg), methylmethan sulfonate (MMS) (80 mg/kg), mitomycin C (MMC) (2 mg/kg) and diethylnitrosamine (DEN) (50 mg/kg) were intraperitoneally dosed to each group consisting of 4 male ddY mice. The mice were subjected to PH 3, 8 or 24 hr after dosing of each carcinogen, and comet assay was performed using the removed liver. The regenerated hepatocyte was sampled five days after PH, and the incidence of micronucleus was measured. CP, MMS, MMC and DEN induced DNA damage at 8 and 24 hr after dosing, while 1-NP induced DNA damage only 8 hr after dosing. All five carcinogens induced micronuclei whenever PH was performed. In the case of CP, the peak of DNA damage was 24 hr after dosing and the timing of PH did not remarkably affect the incidence of micronuclei. The other 4 carcinogens showed peak DNA damage at 8 hr and the highest incidence of micronuclei when PH was operated 24 hr after dosing. In conclusion, we are the first to show the relation of induction between DNA damage and micronucleus in the liver from the same mouse, and tentatively showed the optimal timing of PH as 24 hr after dosing.


Complete Resection of a Cavoatrial Metastatic Liposarcoma under Hypothermic Circulatory Arrest.
A patient underwent surgical resection twice for primary and metastatic dedifferentiated liposarcomas. Computed tomography revealed a tumor mass at the cavoatrial junction. Prompt surgical resection of the tumor with thrombectomy was successfully performed using cardiopulmonary bypass with hypothermic circulatory arrest. Despite the poor prognosis of metastatic or recurrent liposarcoma, the patient has survived for 8 years since the first tumor resection.


Altered Dynamics of Canonical Feedback Inhibition Predicts Increased Burst Transmission in Chronic Epilepsy.
Inhibitory interneurons, organized into canonical feedforward and feedback motifs, play a key role in controlling normal and pathological neuronal activity. We demonstrate prominent quantitative changes in the dynamics of feedback inhibition in a rat model of chronic epilepsy (male Wistar rats). Systematic interneuron recordings revealed a large decrease in intrinsic excitability of basket cells and oriens-lacunosum moleculare interneurons in epileptic animals. Additionally, the temporal dynamics of interneuron recruitment by recurrent feedback excitation were strongly altered, resulting in a profound loss of initial feedback inhibition during synchronous CA1 pyramidal activity. Biophysically constrained models of the complete feedback circuit motifs of normal and epileptic animals revealed that, as a consequence of altered feedback inhibition, burst activity arising in CA3 is more strongly converted to a CA1 output. This suggests that altered dynamics of feedback inhibition promote the transmission of epileptiform bursts to hippocampal projection areas.SIGNIFICANCE STATEMENT We quantitatively characterized changes of the CA1 feedback inhibitory circuit in a model of chronic temporal lobe epilepsy. This study shows, for the first time, that dynamic recruitment of inhibition in feedback circuits is altered and establishes the cellular mechanisms for this change. Computational modeling revealed that the observed changes are likely to systematically alter CA1 input-output properties leading to (1) increased seizure propagation through CA1 and (2) altered computation of synchronous CA3 input.


Cognitive function and the costs of Alzheimer disease. An exploratory study.
To estimate the dollar savings in costs attainable from drug or other treatments for Alzheimer disease (AD) that stabilize or reverse patients' cognitive decline. Medical and other disease-related utilization data were collected from the caregivers of 64 patients diagnosed as having probable AD. The quantities of utilization were priced at national levels to generate measures of illness costs. Costs per patient were then estimated as regression functions of scores on the Mini-Mental State Examination (MMSE), which was used as an index of patient cognitive function. Potential savings in illness costs were estimated by comparing predicted costs at various baseline and intervention-level values of the patient's MMSE score. The potential savings in illness costs attainable from treatment are small for mildly and very severely demented patients with AD. However, for moderately to severely demented home-dwelling patients having, say, an MMSE score of 7 at baseline, prevention of a 2-point decline in the score would save about $3700 annually, and a 2-point increase in an MMSE score rather than a 2-point decline would save about $7100. Large savings in the costs of caring for moderately to severely demented home-dwelling patients with AD may be achievable from disease interventions that have minor effects on patients' cognitive status.


A cluster separation measure.
A measure is presented which indicates the similarity of clusters which are assumed to have a data density which is a decreasing function of distance from a vector characteristic of the cluster. The measure can be used to infer the appropriateness of data partitions and can therefore be used to compare relative appropriateness of various divisions of the data. The measure does not depend on either the number of clusters analyzed nor the method of partitioning of the data and can be used to guide a cluster seeking algorithm.


Shifting visual attention in space: an electrophysiological analysis using high spatial resolution mapping.
Evidence from cortical electrophysiology and functional imaging converges on the view that visual spatial selective attention results in a facilitation of early sensory processing in visual cortical structures. Little is known, however, about the neural control processes that lead to this facilitation. The present study was aimed at further investigating these control processes and their neural correlates by analyzing high spatial resolution maps of brain activity that were evoked by attention-directing cues, but occurred prior to presentation of the target stimulus. Subjects (n=14) were presented with central arrow cues that instructed them to attend covertly to either a left or right field location in order to compare two subsequent target stimuli simultaneously presented to the location. On half of the trials, targets were presented to the cued location, while in the other half, targets were presented to the opposite visual field location. Subjects had to respond via button press on 16% of the trials when target stimuli were identical. Event-related potentials (ERPs) were recorded from 92 scalp electrodes which allowed a sufficiently finegrained analysis of the regional specificity of the ERP components. In response to the cues, an initial component over occipital-parietal electrode sites was consistent with an early involvement of the posterior-parietal cortex, perhaps in the initial step of attentional orienting. A second component over the lateral-prefrontal cortex is consistent with the voluntary control and maintenance of attention, a function known to be subserved by frontal cortical structures. A late component narrowly focussed over occipital-temporal electrode sites is most plausibly related to activation of parts of the ventral extrastriate cortex. The data support the current view that voluntarily orienting visual attention in space leads to top-down modulations in cortical excitability of ventral extrastriate regions initiated by posterior-parietal and mediated by lateral-prefrontal cortical structures.


The role of decay accelerating factor in the immunopathogenesis of cytomegalovirus infection.
A wide variety of the host immune elements play an influential role in the defence against cytomegalovirus (CMV) infection. However, the role of complement in the clearance of CMV infection is less well studied. Decay accelerating factor (DAF/CD55) is a membrane-bound complement regulatory protein that inhibits the formation and accelerates the decay of C3-convertase. Here we hypothesize that murine CMV (MCMV) utilizes DAF as an immunoevasive strategy through down-regulation of host adaptive responses against the virus. To test our hypothesis, DAF knock-out (DAF KO) C57BL/6 mice and wild-type (WT) littermates were infected with a sublethal dose of MCMV, and their immune responses were compared. WT mice lost 7·8% of their initial weight within the first 4 days after infection and quickly began to recover. This is in contrast to the DAF KO mice, that lost a total of 19·4% of their initial weight and did not start recovery until 6 days post-infection. Flow cytometric analysis of lung digests revealed that infected DAF KO mice had a significantly increased infiltration of inflammatory cells, the majority being CD8(+) T lymphocytes. Serum levels of tumour necrosis factor (TNF)-α and interferon (IFN)-γ were also increased markedly in the DAF KO mice compared to the infected WT mice. More interestingly, increased viral genome copies (DNA) in the splenocytes of DAF KO mice was accompanied with mRNA transcripts in the DAF KO mice, an indication of active viral replication. These data suggest an intriguing effect of reduced DAF expression on host responses following in vivo MCMV infection.


Medical Librarianship in Nigeria--a review of the literature and comments on some problems and prospects.
The literature of medical librarianship of Nigeria is reviewed. The staff structure of Nigerian univeristy libraries and, in particular, of the medical libraries attached to them is restrictive, unprogressive, and unconducive to the development of medical librarianship in Nigeria. These medical libraries should cease to be administered and regarded as just unavoidable appendages of the main university libraries. They should be independent, full-fledged libraries of their own, recognixed as full academic departments of their respective colleges or faculties, with their heads being in no way inferior in status to other heads of academic departments. The granting of faculty status to Nigerian unviersity librarians should go the whole way and let the principle of multiple professorships be applied to the staff structure of unviersity libraries. Efforts are being made to effect bibliographic organization of Nigerian medical literature. A national library of medicine for Nigeria, however, humble its beginning, should be established.


Ancestral calling, traditional health practitioner training and mental illness: An ethnographic study from rural KwaZulu-Natal, South Africa.
This qualitative ethnographic study complements an epidemiological study on first episode psychosis in Vulindlela, a rural area in KwaZulu-Natal, South Africa. It focuses on two themes that emerged from our data: (1) the calling of the ancestors to become a traditional health practitioner and (2) ukuthwasa, the training to become a traditional health practitioner. The purpose of this study is to describe the ancestral calling, and to explore whether ukuthwasa may help with the management of mental disturbances, including unusual perceptual experiences. We also provide a discussion of the changing sociopolitical context of healing in KwaZulu-Natal, as a background to our study. In-depth interviews were conducted with 20 (apprentice) traditional health practitioners, formal health practitioners, patients and relatives recruited through local traditional health practitioners and a health care clinic. Our results show that the ancestral calling might announce itself with symptoms of mental illness including unusual perceptual experiences, for which some participants consider ukuthwasa as the only effective cure. We found indications that in some individuals successful completion of ukuthwasa might promote recovery from their illness and lead to a profession in which the unusual perceptual experiences become a legitimate and positively valued aspect. We suggest that - in this particular community today, which has been subject to several sociopolitical changes - ukuthwasa may be a culturally sanctioned healing process which moderates experiences that a Western psychiatric system might characterize as psychotic symptoms, providing some individuals with a lucrative and respected role in society.


[Neuronal activity of cat thalamus reticular and relay nuclei].
The interaction between the unit activity of the thalamic reticular and relay nuclei was investigated on cats immobilized with tubarine. It was shown that unit activity of the relay nuclei can be significantly modulated by stimulation of the n. reticul. By simultaneous recording (with two microelectrodes) of the activity of thalamic reticular neurons the relay unit activity can be suppressed; during peripheral and cortical stimulations the alternation of excitation-inhibition of the thalamic reticular and relay units can be observed; excitation of thalamic reticular units can be associated with excitation of the relay nuclei units. Such types of interaction may also exist during simultaneous recording (by two microelectrodes) of the activity of the relay units and putative interneurons. Sometimes IPSPs with short latency (1 ms) were evoked in the thalamic relay nuclei during thalamic reticular stimulation. The existence of direct monosynaptic inhibition of the relay unit activity by the thalamic reticular units as well as inhibition by the activation of the putative interneurons of the specific thalamic nuclei is suggested.


Glia and pain: is chronic pain a gliopathy?
Activation of glial cells and neuro-glial interactions are emerging as key mechanisms underlying chronic pain. Accumulating evidence has implicated 3 types of glial cells in the development and maintenance of chronic pain: microglia and astrocytes of the central nervous system (CNS), and satellite glial cells of the dorsal root and trigeminal ganglia. Painful syndromes are associated with different glial activation states: (1) glial reaction (ie, upregulation of glial markers such as IBA1 and glial fibrillary acidic protein (GFAP) and/or morphological changes, including hypertrophy, proliferation, and modifications of glial networks); (2) phosphorylation of mitogen-activated protein kinase signaling pathways; (3) upregulation of adenosine triphosphate and chemokine receptors and hemichannels and downregulation of glutamate transporters; and (4) synthesis and release of glial mediators (eg, cytokines, chemokines, growth factors, and proteases) to the extracellular space. Although widely detected in chronic pain resulting from nerve trauma, inflammation, cancer, and chemotherapy in rodents, and more recently, human immunodeficiency virus-associated neuropathy in human beings, glial reaction (activation state 1) is not thought to mediate pain sensitivity directly. Instead, activation states 2 to 4 have been demonstrated to enhance pain sensitivity via a number of synergistic neuro-glial interactions. Glial mediators have been shown to powerfully modulate excitatory and inhibitory synaptic transmission at presynaptic, postsynaptic, and extrasynaptic sites. Glial activation also occurs in acute pain conditions, and acute opioid treatment activates peripheral glia to mask opioid analgesia. Thus, chronic pain could be a result of "gliopathy," that is, dysregulation of glial functions in the central and peripheral nervous system. In this review, we provide an update on recent advances and discuss remaining questions.


Biosafety of E. coli beta-glucuronidase (GUS) in plants.
The beta-glucuronidase (GUS) gene is to date the most frequently used reporter gene in plants. Marketing of crops containing this gene requires prior evaluation of their biosafety. To aid such evaluations of the GUS gene, irrespective of the plant into which the gene has been introduced, the ecological and toxicological aspects of the gene and gene product have been examined. GUS activity is found in many bacterial species, is common in all tissues of vertebrates and is also present in organisms of various invertebrate taxa. The transgenic GUS originates from the enterobacterial species Escherichia coli that is widespread in the vertebrate intestine, and in soil and water ecosystems. Any GUS activity added to the ecosystem through genetically modified plants will be of no or minor influence. Selective advantages to genetically modified plants that posses and express the E. coli GUS transgene are unlikely. No increase of weediness of E. coli GUS expressing crop plants, or wild relatives that might have received the transgene through outcrossing, is expected. Since E. coli GUS naturally occurs ubiquitously in the digestive tract of consumers, its presence in food and feed from genetically modified plants is unlikely to cause any harm. E. coli GUS in genetically modified plants and their products can be regarded as safe for the environment and consumers.


C57BL/10/CR mice: nonresponders to activation by the lipid a moiety of bacterial lipopolysaccharide.
C57BL/10/CR mice do not respond to the lipid A moiety of LPS. This defect was demonstrated in vivo by a decreased production of the acute phase serum amyloid protein SAA. In addition, the defect was demonstrated in vitro by using cultures of spleen cells and peritoneal cells. No mitogenesis of spleen cells or enhanced glucose utilization by either spleen cells or peritoneal cells was observed when the cells were stimulated by lipid A or phenol-extracted Escherichia coli K235 LPS. The response of these mice to PHA, Con A, Poly I:C, 8BrcGMP, and butanol extraced E. coli K235 LPS was normal. Thus, the inability of lipid A to stimulate B lymphocyte mitogenesis and activate peritoneal cells in vitro may correlate with its inability to induce the acute phase SAA in vivo. B10/CR mice represent another strain, similar to the C3H/HeJ, which is nonresponsive to lipid A and should be useful in eludcidating the mechanism by which bacterial LPS activates cells.


Repair after nephron ablation reveals limitations of neonatal neonephrogenesis.
The neonatal mouse kidney retains nephron progenitor cells in a nephrogenic zone for 3 days after birth. We evaluated whether de novo nephrogenesis can be induced postnatally beyond 3 days. Given the long-term implications of nephron number for kidney health, it would be useful to enhance nephrogenesis in the neonate. We induced nephron reduction by cryoinjury with or without contralateral nephrectomy during the neonatal period or after 1 week of age. There was no detectable compensatory de novo nephrogenesis, as determined by glomerular counting and lineage tracing. Contralateral nephrectomy resulted in additional adaptive healing, with little or no fibrosis, but did not also stimulate de novo nephrogenesis. In contrast, injury initiated at 1 week of age led to healing with fibrosis. Thus, despite the presence of progenitor cells and ongoing nephron maturation in the newborn mouse kidney, de novo nephrogenesis is not inducible by acute nephron reduction. This indicates that additional nephron progenitors cannot be recruited after birth despite partial renal ablation providing a reparative stimulus and suggests that nephron number in the mouse is predetermined at birth.


Influence of pregnancy and lactation on maternal deposition and perinatal uptake of 241Am in the rat.
To investigate the metabolism of 241Am as affected by pregnancy and lactation, female rats were injected with 5 muCi of 241Am intravenously while nulliparous, pregnant or lactating. The females and subsequent litters were killed at various times after injection to determine 241Am distribution and retention. The temporal relationship between injection and pregnancy influenced the tissue retention of 241Am in both dams and progeny. The half-time of 241Am in livers of pregnant or lactating rats was more than twice that of nulliparous rats, although the initial uptake was approximately 50% of the activity in all groups. Both spleen and femur accumulated more 241Am at 7-10 weeks after injection than at earlier times, but this increase could not be related to pregnancy. Approximately 10 times more 241Am was transferred to offspring from dams that were lactating when exposed than was transferred via the placenta when exposure occurred late in gestation. Furthermore, more 241Am was transmitted to the progeny via milk if exposure of the dam occurred during lactation rather than during pregnancy.


Outcome for children with supratentorial primitive neuroectodermal tumors treated with surgery, radiation, and chemotherapy.
The outcome of a child with a primitive neuroectodermal tumors arising supratentorially (SPNET) is not well characterized and may differ from the outcome of a patient with a histologically similar cerebellar tumor (medulloblastoma [MB]). Recently, 5-year progression free survival rates as high as 80% have been reported for children with MB treated with craniospinal radiation (CRT) and chemotherapy including cisplatin, lomustine (CCNU), and vincristine (VCR). The authors reviewed the outcome of 22 consecutive patients age 3 years and older (mean age, 10 years; range, 3-18 years) with SPNET who were treated at the study institutions between 1981 and 1996. Tumor location included was 13 pineal, 6 cortical, and 3 thalamic or suprasellar. Five patients had disease dissemination at diagnosis. All patients underwent surgery and staging, followed by CRT and chemotherapy with cisplatin, CCNU, and VCR. Of the 22 patients, 13 had developed disease progression and 10 had died at the time of last follow-up. Overall progression free survival (PFS) was 47% +/- 11% at 3 years and 37% +/- 11% at 5 years. There was a significant difference in PFS between patients with localized disease versus those with disseminated disease (P = 0.04). There was no statistical association between tumor location and survival. Although not significant (P = 0.21), there was a trend toward better survival of those patients with complete or near-complete resection compared with those with partial resection or biopsy. The results of the current study demonstrate that the outcome for children with SPNET treated with radiation and chemotherapy appears worse than for children with MB treated with identical therapy. This suggests that there may be biologic differences between supratentorial and infratentorial primitive neuroectodermal tumors, thus requiring refinements in treatment.


Telemedicine Evaluations in Neuro-Ophthalmology During the COVID-19 Pandemic: Patient and Physician Surveys.
The novel coronavirus 2019 (COVID-19) pandemic has transformed health care. With the need to limit COVID-19 exposures, telemedicine has become an increasingly important format for clinical care. Compared with other fields, neuro-ophthalmology faces unique challenges, given its dependence on physical examination signs that are difficult to elicit outside the office setting. As such, it is imperative to understand both patient and provider experiences to continue to adapt the technology and tailor its application. The purpose of this study is to analyze both neuro-ophthalmology physician and patient satisfaction with virtual health visits during the time of the COVID-19 pandemic. Across three institutions (NYU Langone Health, Indiana University Health, and Columbia University Medical Center), telemedicine surveys were administered to 159 patients. Neuro-ophthalmologists completed 157 surveys; each of these were linked to a single patient visit. Patient surveys consisted of 5 questions regarding visit preparation, satisfaction, challenges, and comfort. The physician survey included 4 questions that focused on ability to gather specific clinical information by history and examination. Among 159 patients, 104 (65.4%) reported that they were satisfied with the visit, and 149 (93.7%) indicated that they were comfortable asking questions. Sixty-eight (73.9%) patients found the instructions provided before the visit easy to understand. Potential areas for improvement noted by patients included more detailed preparation instructions and better technology (phone positioning, Internet connection, and software). More than 87% (137/157) of neuro-ophthalmologists surveyed reported having performed an examination that provided enough information for medical decision-making. Some areas of the neuro-ophthalmologic examination were reported to be easy to conduct (range of eye movements, visual acuity, Amsler grids, Ishihara color plates, and pupillary examination). Other components were more difficult (saccades, red desaturation, visual fields, convergence, oscillations, ocular alignment, and smooth pursuit); some were especially challenging (vestibulo-ocular reflex [VOR], VOR suppression, and optokinetic nystagmus). Clinicians noted that virtual health visits were limited by patient preparation, inability to perform certain parts of the examination (funduscopy and pupils), and technological issues. Among virtual neuro-ophthalmology visits evaluated, most offer patients with appointments that satisfy their needs. Most physicians in this cohort obtained adequate clinical information for decision-making. Even better technology and instructions may help improve aspects of virtual health visits.


Management of dry eye disease.
The management of dry eye disease (DED) encompasses both pharmacologic and nonpharmacologic approaches, including avoidance of exacerbating factors, eyelid hygiene, tear supplementation, tear retention, tear stimulation, and anti-inflammatory agents. Artificial tears are the mainstay of DED therapy but, although they improve symptoms and objective findings, there is no evidence that they can resolve the underlying inflammation in DED. Topical corticosteroids are effective anti-inflammatory agents, but are not recommended for long-term use because of their adverse-effect profiles. Topical cyclosporine--currently the only pharmacologic treatment approved by the US Food and Drug Administration specifically for DED--is safe for long-term use and is disease-modifying rather than merely palliative. Treatment selection is guided primarily by DED severity. Recently published guidelines propose a severity classification based on clinical signs and symptoms, with treatment recommendations according to severity level.


[Cloning of bacteriophage T5 DNA fragments in the plasmid pBR322. Analysis of recombinant plasmids by the method of bonding with RNA-polymerase from Escherichia coli on nitrocellulose filters].
DNA of bacteriophage T5 was hydrolyzed with restriction endonucleases HindIII and BamHI, and subjected to the combined hydrolysis with BamHI+EcoRI and BamHI+ +HindIII. Fragments obtained were cloned in the plasmid pBR322. About 17% of T5 genome were recovered in recombinant plasmids. Cloned fragments were localized on the physical map of the phage by restriction analysis and Southern hybridization. With the aim of direct cloning of T5 promoters, PstI/HindIII fragments were inserted into pBR322 followed by selection of recombinants on ApsTCr phenotype. Binding of BsuRI and AluI fragments of hybrid plasmids with E. coli RNA polymerase was studied by nitrocellulose filter assay. The fragments, which were capable to form heparin resistant complexes were identified.


Fosfomycin: Pharmacological, Clinical and Future Perspectives.
Fosfomycin is a bactericidal, low-molecular weight, broad-spectrum antibiotic, with putative activity against several bacteria, including multidrug-resistant Gram-negative bacteria, by irreversibly inhibiting an early stage in cell wall synthesis. Evidence suggests that fosfomycin has a synergistic effect when used in combination with other antimicrobial agents that act via a different mechanism of action, thereby allowing for reduced dosages and lower toxicity. Fosfomycin does not bind to plasma proteins and is cleared via the kidneys. Due to its extensive tissue penetration, fosfomycin may be indicated for infections of the CNS, soft tissues, bone, lungs, and abscesses. The oral bioavailability of fosfomycin tromethamine is <50%; therefore, oral administration of fosfomycin tromethamine is approved only as a 3-gram one-time dose for treating urinary tract infections. However, based on published PK parameters, PK/PD simulations have been performed for several multiple-dose regimens, which might lead to the future use of fosfomycin for treating complicated infections with multidrug-resistant bacteria. Because essential pharmacological information and knowledge regarding mechanisms of resistance are currently limited and/or controversial, further studies are urgently needed, and fosfomycin monotherapy should be avoided.


Pittosporum cryptic virus 1: genome sequence completion using next-generation sequencing.
Next-generation sequencing (NGS) was applied to dsRNAs extracted from an Italian pittosporum plant infected with pittosporum cryptic virus 1 (PiCV1). NGS allowed assembly of the full genome sequence of PiCV1, comprising dsRNA1 (1.9 kbp) and dsRNA2 (1.5 kbp), which encode the RNA-dependent RNA polymerase and capsid protein genes, respectively. Phylogenetic and sequence analyses confirmed that PiCV1 is a new member of the genus Deltapartitivirus, family Partiviridae. From the same plant, NSG also permitted assembly of the complete genome sequence of eggplant mottled dwarf virus (EMDV), which shared 86 % to 98 % nucleotide sequence identity with complete and partial sequences (ca 6750 nt) of other known EMDV isolates with sequences available in the GenBank database.


Naturally occurring opioid receptor agonists stimulate adenylate cyclase activity in rat olfactory bulb.
In homogenate of rat olfactory bulb, the opioid receptor agonists beta-endorphin, Leu-enkephalin, and dynorphin A stimulated adenylate cyclase activity in a concentration-dependent manner, with half-maximal effects displayed at 22, 63, and 176 nM, respectively. The maximal stimulation of the enzyme activity corresponded to about a 40% increase of basal activity for all three peptides. Naloxone antagonized the stimulation of beta-endorphin, Leu-enkephalin, and dynorphin A, with pA2 values of 8.0, 7.7, and 8.1, respectively. Kinetic analysis performed with Leu-enkephalin showed that the opioid peptide increased the Vmax of the enzyme, without changing the Km for the substrate Mg-ATP. Moreover, the opioid stimulation was associated with a significant increase of the affinity of the enzyme for Mg2+ activation and occurred in membranes incubated in a Ca2(+)-free medium. Addition of exogenous GTP at micromolar concentrations was absolutely necessary for the detection of the opioid effect. Treatment of olfactory bulbs with cholera toxin did not alter the stimulation of adenylate cyclase by Leu-enkephalin. However, the opioid stimulation disappeared in membranes obtained from bulbs injected with pertussis toxin. These results demonstrate the presence in the brain of a new functional class of opiate receptors coupled to stimulation of adenylate cyclase via a transduction mechanism that is Ca2+ independent and seems to involve a pertussis toxin-sensitive GTP-binding protein.


Evidence for transcriptional regulation of plastid photosynthesis genes in Arabidopsis thaliana roots.
Mechanisms underlying suppressed levels of transcripts for plastid photosynthesis genes in nongreen tissues such as roots and calli were analyzed in Arabidopsis thaliana, a plant suitable for further genetic dissection. A region encoding promoters of rbcL, the gene encoding the large subunit of ribulose-1,5-biphosphate carboxylase/oxygenase, and the atpB/E operon for the beta and epsilon subunits of coupling factor one were cloned and sequenced. Transcripts for rbcL, atpB/E, and psbA, the gene for the D1 protein in the photosystem II reaction center, were barely detectable in roots of A. thaliana, whereas 16S rRNA was detected at a low level. The run-on transcription experiment revealed that expression of rbcL, atpB/E, and psbA was regulated at transcription. The copy number of plastid DNA in roots was one-fifth that in green leaves on the basis of total cellular DNA, suggesting that in the latter the DNA copy-number regulation also exists in plastid gene expression. Digestion of DNA with methyl-sensitive and -insensitive isoschizomeric endonucleases and subsequent polymerase chain reaction, as well as in vitro transcription of plastid DNAs with Escherichia coli RNA polymerase, resulted in no evidence of regulation by DNA modification. In spite of predominant suppression of expression of rbcL, atpB/E, and psbA at transcription in roots and calli, 16S rRNA levels were decreased because of low RNA stability.


In search of biomarkers in psychiatry: EEG-based measures of brain function.
Current clinical parameters used for diagnosis and phenotypic definitions of psychopathology are both highly variable and subjective. Intensive research efforts for specific and sensitive biological markers, or biomarkers, for psychopathology as objective alternatives to the current paradigm are ongoing. While biomarker research in psychiatry has focused largely on functional neuroimaging methods for identifying the neural functions that associate with psychopathology, scalp electroencephalography (EEG) has been viewed, historically, as offering little specific brain source information, as scalp appearance is only loosely correlated to its brain source dynamics. However, ongoing advances in signal processing of EEG data can now deliver functional EEG brain-imaging with distinctly improved spatial, as well as fine temporal, resolution. One computational approach proving particularly useful for EEG cortical brain imaging is independent component analysis (ICA). ICA decomposition can be used to identify distinct cortical source activities that are sensitive and specific to the pathophysiology of psychiatric disorders. Given its practical research advantages, relatively low cost, and ease of use, EEG-imaging is now both feasible and attractive, in particular for studies involving the large samples required by genetically informative designs to characterize causal pathways to psychopathology. The completely non-invasive nature of EEG data acquisition, coupled with ongoing advances in dry, wireless, and wearable EEG technology, makes EEG-imaging increasingly attractive and appropriate for psychiatric research, including the study of developmentally young samples. Applied to large genetically and developmentally informative samples, EEG imaging can advance the search for robust diagnostic biomarkers and phenotypes in psychiatry.


Synthesis of Poly[APMA]-DOTA-64Cu conjugates for interventional radionuclide therapy of prostate cancer: assessment of intratumoral retention by micro-positron emission tomography.
To develop new radiopharmaceuticals for interventional radionuclide therapy of locally recurrent prostate cancer, poly[N-(3-aminopropyl)methacrylamide] [poly(APMA)] polymers were synthesized by free radical precipitation polymerization in acetone-dimethylsulfoxide using N,N'-azobis(isobutyronitrile) as the initiator. The polymers were characterized with nuclear magnetic resonance, size exclusion chromatography, and dynamic light scattering (M(n) = 2.40 x 10(4), M(w)/M(n) = 1.87). Subsequently, poly[APMA] was coupled with 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride as an activator, followed by conjugation with (64)Cu radionuclide. Prolonged retention of poly[APMA]-DOTA-(64)Cu conjugates within the tumor tissues was demonstrated by micro-positron emission tomography at 24 hours following intra-tumoral injection of the conjugates to human prostate xenografts in mice. The data suggest that the poly[APMA]-DOTA-(64)Cu conjugates might be useful for interventional radionuclide therapy of locally recurrent prostate cancer in humans.


An Epidemiological Model for Examining Marijuana Use over the Life Course.
Trajectories of drug use are usually studied empirically by following over time persons sampled from either the general population (most often youth and young adults) or from heavy or problematic users (e.g., arrestees or those in treatment). The former, population-based samples, describe early career development, but miss the years of use that generate the greatest social costs. The latter, selected populations, help to summarize the most problematic use, but cannot easily explain how people become problem users nor are they representative of the population as a whole. This paper shows how microsimulation can synthesize both sorts of data within a single analytical framework, while retaining heterogeneous influences that can impact drug use decisions over the life course. The RAND Marijuana Microsimulation Model is constructed for marijuana use, validated, and then used to demonstrate how such models can be used to evaluate alternative policy options aimed at reducing use over the life course.


Signatures of Superradiance as a Witness to Multipartite Entanglement.
Generation and detection of entanglement is at the forefront of most quantum information technologies. There is a plethora of techniques that reveal entanglement on the basis of only partial information about the underlying quantum state, including entanglement witnesses. Superradiance refers to the phenomenon of highly synchronized photon emission from an ensemble of quantum emitters that is caused by correlations among the individual particles and has been connected by Dicke himself to the presence of multipartite entangled states. We investigate this connection in a quantitative way and discuss whether or not signatures of superradiance from semiconductor nanolasers, manifesting themselves as a modification of the spontaneous-emission time, can be interpreted as a witness to detect entanglement in the underlying state of the emitters.


Analysis of NZB hyperdiploid spleen cells.
In order to study the cell types involved in the hyperdiploidy characteristic of NZB mice, flow cytometric techniques, which measure te DNA content of individual cells, have been used together with standard cytogenetic analysis. The aneuploid cells present in NZB mice were found to be clonally derived. These cells were of large size relative to nonaneuploid cells. They could not be removed by lysis with anti-Thy 1 plus complement nor were they present in the nylon-wool column nonadherent fraction, both of which are characteristic of T cells. By further cytotoxic analysis, the aneuploid cells were found not to express Ly-5 nor NK surface antigens, found on natural killer cells. The aneuploid cells had increased quantities of cell surface H-2 antigen. This marked susceptibility to lysis with anti-H-2 serum was associated with one or more extra copies of chromosome 17, which carries the H-2 complex. A large proportion of these cells also expressed surface Ia and Ig. The aneuploid cells found in the spleens of older NZB mice derive from bone marrow stem cells. Neonatally thymectomized and lethally irradiated NZB X DBA/2 recipients of anti-Thy-1 treated young NZB bone marrow cells ultimately developed aneuploid spleen cells. Such cells are not found in intact or thymectomized NZB Z DBA/2 mice nor in recipients of DBA/2 marrow. Finally, congenic NZB mice carrying the CBZ/N xid failed to develop aneuploidy. Taken together, these studies suggest that the aneuploid cell is a bone marrow-derived stem cell destined to differentiate into the B cell subset lacking in CBA/N mice.


Trimeric autotransporter adhesin-dependent adherence of Bartonella henselae, Bartonella quintana, and Yersinia enterocolitica to matrix components and endothelial cells under static and dynamic flow conditions.
Trimeric autotransporter adhesins (TAAs) are important virulence factors of Gram-negative bacteria responsible for adherence to extracellular matrix (ECM) and host cells. Here, we analyzed three different TAAs (Bartonella adhesin A [BadA] of Bartonella henselae, variably expressed outer membrane proteins [Vomps] of Bartonella quintana, and Yersinia adhesin A [YadA] of Yersinia enterocolitica) for mediating bacterial adherence to ECM and endothelial cells. Using static (cell culture vials) and dynamic (capillary flow chambers) experimental settings, adherence of wild-type bacteria and of the respective TAA-negative strains was analyzed. Under static conditions, ECM adherence of B. henselae, B. quintana, and Y. enterocolitica was strongly dependent on the expression of their particular TAAs. YadA of Y. enterocolitica did not mediate bacterial binding to plasma or cellular fibronectin under either static or dynamic conditions. TAA-dependent host cell adherence appeared more significant under dynamic conditions although the total number of bound bacteria was diminished compared to the number under static conditions. Dynamic models expand the methodology to perform bacterial adherence experiments under more realistic, bloodstream-like conditions and allow dissection of the biological role of TAAs in ECM and host cell adherence under static and dynamic conditions.


Automated phase characterization and adaptive pulse compression using multiphoton intrapulse interference phase scan in air.
We introduce a non-interferometric single beam method for automated spectral phase characterization and adaptive pulse compression of amplified ultrashort femtosecond pulses taking advantage of third order harmonic generation in air. This new method, air-MIIPS, compensates high-order phase distortions based on multiphoton intrapulse interference phase scan (MIIPS).


Prevention and control of mousepox.
The prevention and control of mousepox begins with investigators being aware of the disease's existence and its serious nature for users of mice and mouse tissues. Every institution must include as part of its management policy, provisions to ascertain that mice and mouse tissues entering its facilities are free of ectromelia virus. Policy must encompass the entire institution, be clearly stated, and must be workable. The provisions of this policy include the establishment of a disease surveillance program which actively searches for disease or virus in incoming mice and mouse tissues from untested sources. Complete records of receipts of animals and animal tissues must be maintained. There must be open and honest communication among investigators concerning the presence of mousepox (and other diseases) in their mice and tumors. Decisions must be based on accurate information. The elimination of mousepox within an institution may be difficult, and a single plan applicable to all situations cannot be given. However, the basics of all control measures include the following: recognition that mousepox represents an institutional problem, not the problem of only a single or a few investigators; rapid notification of all investigators involved, including those who may have received infected mice; confirmation of the diagnosis and notification of the scientific community of the disease's existence. Strategies for dealing with mousepox depend on the nature of the mice infected. Euthanasia of mice which can be replaced and vaccination of extremely valuable breeding stock are suggested.


Diversity of protein and mRNA forms of mammalian methionine sulfoxide reductase B1 due to intronization and protein processing.
Methionine sulfoxide reductases (Msrs) are repair enzymes that protect proteins from oxidative stress by catalyzing stereospecific reduction of oxidized methionine residues. MsrB1 is a selenocysteine-containing cytosolic/nuclear Msr with high expression in liver and kidney. Here, we identified differences in MsrB1 gene structure among mammals. Human MsrB1 gene consists of four, whereas the corresponding mouse gene of five exons, due to occurrence of an additional intron that flanks the stop signal and covers a large part of the 3'-UTR. This intron evolved in a subset of rodents through intronization of exonic sequences, whereas the human gene structure represents the ancestral form. In mice, both splice forms were detected in liver, kidney, brain and heart with the five-exon form being the major form. We found that both mRNA forms were translated and supported efficient selenocysteine insertion into MsrB1. In addition, MsrB1 occurs in two protein forms that migrate as 14 and 5 kDa proteins. We found that each mRNA splice form generated both protein forms. The abundance of the 5 kDa form was not influenced by protease inhibitors, replacement of selenocysteine in the active site or mutation of amino acids in the cleavage site. However, mutation of cysteines that coordinate a structural zinc decreased the levels of 5 and 14 kDa forms, suggesting importance of protein structure for biosynthesis and/stability of these forms. This study characterized unexpected diversity of protein and mRNA forms of mammalian selenoprotein MsrB1.


Aging and luteinizing hormone effects on reactive oxygen species production and DNA damage in rat Leydig cells.
We observed previously that after long-term suppression of luteinizing hormone (LH) and thus of Leydig cell steroidogenesis, restimulation of the Leydig cells by LH resulted in significantly higher testosterone production than by age-matched cells from control rats. These studies suggest that stimulation over time may elicit harmful effects on the steroidogenic machinery, perhaps through alteration of the intracellular oxidant-to-antioxidant balance. Herein we compared the effects of LH stimulation on stress response genes, formation of intracellular reactive oxygen species (ROS), and ROS-induced damage to ROS-susceptible macromolecules (DNA) in young and in aged cells. Microarray analysis indicated that LH stimulation resulted in significant increases in expression of genes associated with stress response and antiapoptotic pathways. Short-term LH treatment of primary Leydig cells isolated from young rats resulted in transiently increased ROS levels compared to controls. Aged Leydig cells also showed increased ROS soon after LH stimulation. However, in contrast to the young cells, ROS production peaked later and the time to recovery was increased. In both young and aged cells, treatment with LH resulted in increased levels of DNA damage but significantly more so in the aged cells. DNA damage levels in response to LH and the levels of intracellular ROS were highly correlated. Taken together, these results indicate that LH stimulation causes increased ROS production by young and aged Leydig cells and that while DNA damage occurs in cells of both ages, there is greater damage in the aged cells.


Slow magic-angle coil spinning: a high-sensitivity and high-resolution NMR strategy for microscopic biological specimens.
High-resolution magic-angle spinning has become a valuable tool to study biologic samples; however, there are only a few high-resolution magic-angle spinning methodologies that have been explored specifically to deal with small semisolid biomaterials such as tissues. Here, we report a novel high-resolution (1)H NMR spectroscopic approach using magic-angle coil spinning. We demonstrate its potential for intact tissues by combining a resonant microcoil with a large-volume commercial magic-angle spinning rotor, where the microcoil offers incredible sensitivity and the large-volume magic-angle spinning rotor provides stable slow magic-angle spinning. This approach provides high-resolution spectra with high sensitivity and the preservation of living biologic specimens for an accurate chemical analysis.


Efficacy of Yavakshara Taila Uttarabasti in the management of fallopian tube blockage.
Tubal blockage is one of the most common causative factors for female barrenness. It accounts for about 25-35% of female infertility. It is very difficult to manage, as the treatment choices for it are only tubal re-constructive surgery and in vitro fertilization (IVF). On the other hand, there is not established any reliable Ayurvedic treatment for the tubal blockage. It is the need of the time to establish an efficient and cost-effective therapy for this problem. To evaluate the efficacy of Yavakshara Taila Uttarabasti in fallopian tubal blockage. Patients of childbearing age with active marital life of 1 year or more, having complaint of failure to conceive with at least one fallopian tube blockage were selected. Total 19 patients were registered with 42.11% unilateral and 57.89% bilateral tubal blockage. Yavakshara Taila (5 ml) Intrauterine Uttarabasti was given for 6 days (with interval of 3 days in between), after completion of menstrual cycle for two consecutive cycles. The tubal patency was found in 68.75% of patients and conception was achieved in 6.25% of patients. Yavakshara Taila Uttarabasti an effective procedure for treating tubal blockage with no apparent evidence of complication.


Acute effects of ANP and BNP on hypoxic pulmonary vasoconstriction in humans.
1. Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) have pulmonary vasorelaxant activity with plasma concentrations being elevated in patients with hypoxaemic pulmonary hypertension. However, their effects on acute hypoxic pulmonary vasoconstriction (HPV), the initiating stimulus for pulmonary hypertension have not to date been investigated. We have therefore studied the effects of ANP and BNP on acute HPV in humans. 2. Eight healthy volunteers were studied on three separate occasions. After reaching a resting haemodynamic state (t0), an infusion of either ANP (10 pmol kg-1 min-1), BNP (10 pmol kg-1 min-1) or placebo (5% dextrose) was commenced. This was given alone for 30 min (t30) before subjects were rendered hypoxaemic (SaO2 75-80%) for a further 30 min (t60), with the initial infusion continuing to t60. Pulsed-wave Doppler analysis of pulmonary artery flow was used to measure mean pulmonary arterial pressure (MPAP) and hence total pulmonary vascular resistance (PVR) was calculated. 3. MPAP and PVR both tended to decrease in response to ANP and BNP infusion, although compared with placebo, the difference at t30 was only statistically significant for PVR. Hypoxaemia increased MPAP and PVR, although values at t60 were significantly lower following both ANP and BNP compared with placebo. 4. In terms of the actual change in PVR (delta PVR) induced by hypoxaemia (from t30 to t60), BNP (146(16) dyn s cm-5), but not ANP (183(21) dyn s cm-5) significantly attenuated delta PVR compared with placebo (194(26) dyns s cm-5): mean difference BNP versus placebo 48 dyn s cm-5, 95% Cl 3-93. An identical pattern was observed for delta MPAP where BNP (15.9(1.1) mmHg), but not ANP (18.0(1.2) mmHg) significantly attenuated delta MPAP compared with placebo (19.0(1.7) mmHg): mean difference BNP versus placebo 3.1 mmHg, 95% Cl 0.7-5.5. 5. Thus, although both ANP and BNP exhibit pulmonary vasorelaxant activity, only BNP significantly attenuated the MPAP and PVR responses to acute hypoxaemia. This suggests that the natriuretic peptides may have a role in attenuating pulmonary hypertension secondary to hypoxaemia.


Stabilization of the B-type natriuretic peptide mRNA in cardiac myocytes by alpha-adrenergic receptor activation: potential roles for protein kinase C and mitogen-activated protein kinase.
In cardiac myocytes, B-type natriuretic peptide (BNP) expression is induced with the rapid kinetics of a primary response gene. Like many other primary response gene transcripts, the BNP mRNA possesses destabilizing elements and is believed to be short-lived. The rapid induction of a short-lived transcript could be achieved partty by agonist-mediated increases in mRNA t1/2. Accordingly, the present study was undertaken to evaluate whether the alpha 1-adrenergic receptor agonist, phenylephrine (PE), a known inducer of BNP expression, could stabilize the BNP mRNA and, if so, what signaling pathways might be involved. In primary myocardial cells treated with a transcription inhibitor, the t1/2 of the BNP mRNA was found to be about 1 h in the absence of PE; however, in the presence of PE, the t1/2 increased to 5 h. It was shown that neither the calmodulin kinase inhibitor, KN-62, nor the protein tyrosine kinase inhibitor, tyrphostin, blocked PE-mediated stabilization of the BNP mRNA. However, either the protein kinase C (PKC) inhibitor, GF 109203X, or the mitogen-activated protein kinase kinase (MAPKK) inhibitor, PD 098059, effected some blockade of the stabilizing effects of PE. While maximal doses of PD 098059 nearly completely blocked PE-activated MAPK, stabilization was only partially inhibited. Moreover, maximal doses of GF 109203X, which only partially blocked PE-activated MAPK, nearly completely inhibited stabilization. Thus, while MAPK appears to be required for maximal agonist-mediated stabilization, PKC seems to play a dominant role, participating through both MAPK-dependent and -independent pathways. These results establish roles for both the PKC and MAPK families in alpha 1-adrenergic receptor-mediated stabilization of the BNP mRNA, suggesting that the rapid induction of BNP expression might be due, in part, to this agonist-mediated increase in mRNA t1/2.


Treating restless legs syndrome in the context of sleep disordered breathing comorbidity.
Obstructive sleep apnoea (OSA) and restless legs syndrome (RLS) are two of the most prevalent sleep disorders and can coexist within the same patient. Nonetheless, the recognition of RLS among OSA patients has important clinical implications, since RLS can disrupt sleep despite adequate treatment of sleep disordered breathing and should be treated accordingly. Furthermore, the presence of OSA can also increase the severity of RLS. Therefore, it is important to be able to correctly identify both disorders and treat them effectively. The present article reviews our current knowledge on this comorbidity and discusses potential treatment options for RLS in the context of OSA.


Effectiveness of different local actions to control vitamin D prescription in Italy.
In the last decade, the significant expenditure and consumption increase of vitamin D in Italy led some regions to adopt strategies to improve prescribing appropriateness and contain expenditure. Using the statistical analysis method of interrupted time series for consumption and expenditure of cholecalciferol, different types of interventions adopted in four Italian regions and their efficacy were evaluated. Molise achieved the best results by adopting a health professionals' education program in addition to a prescriber-sanction system. Emilia-Romagna also opted for a medical education strategy, but the results were less relevant due to the lack of penalties. Lazio obtained a slowdown in consumption growth by targeting on the utilization of lower-cost per defined daily dose (DDD) packs and adopting a therapeutic plan. Sardinia showed a decrease in expenditure by adopting a target threshold of lower-cost formulation. The reimbursement of the lowest-cost packs within the National Health Service (NHS) undoubtedly influences spending trend, but it does not solve prescriptive inappropriateness.


Prevention of major amputations in the diabetic patient.
Five of six major amputations of the lower extremity involve diabetic patients. It should be possible to reduce the number of major amputations by substituting reconstructive for destructive surgery, by radical local surgical debridement, by achieving healing of chronic foot lesions, by the early diagnosis of spread of infections from foot to leg, by limiting minor amputations, by improving the blood supply, and by providing a continuum of care by experienced personnel who supervise the foot status of the patient on an ongoing basis. In my experience, the application of these principles over the past seven years has precluded major amputation in the treatment of 48 diabetic patients with serious lower extremity lesions.


Clustrophile 2: Guided Visual Clustering Analysis.
Data clustering is a common unsupervised learning method frequently used in exploratory data analysis. However, identifying relevant structures in unlabeled, high-dimensional data is nontrivial, requiring iterative experimentation with clustering parameters as well as data features and instances. The number of possible clusterings for a typical dataset is vast, and navigating in this vast space is also challenging. The absence of ground-truth labels makes it impossible to define an optimal solution, thus requiring user judgment to establish what can be considered a satisfiable clustering result. Data scientists need adequate interactive tools to effectively explore and navigate the large clustering space so as to improve the effectiveness of exploratory clustering analysis. We introduce Clustrophile 2, a new interactive tool for guided clustering analysis. Clustrophile 2 guides users in clustering-based exploratory analysis, adapts user feedback to improve user guidance, facilitates the interpretation of clusters, and helps quickly reason about differences between clusterings. To this end, Clustrophile 2 contributes a novel feature, the Clustering Tour, to help users choose clustering parameters and assess the quality of different clustering results in relation to current analysis goals and user expectations. We evaluate Clustrophile 2 through a user study with 12 data scientists, who used our tool to explore and interpret sub-cohorts in a dataset of Parkinson's disease patients. Results suggest that Clustrophile 2 improves the speed and effectiveness of exploratory clustering analysis for both experts and non-experts.


Sugar-feeding behaviour and longevity of European Culicoides biting midges.
Most haematophagous insect vectors can also use sugar as an energy source; thus their sugar-feeding behaviour influences their longevity and blood-feeding rate and hence their vectorial capacity. Scant information is available on the sugar-feeding behaviour of Culicoides Latreille biting midges (Diptera: Ceratopogonidae), which are vectors of bluetongue and Schmallenberg viruses. The longevity of laboratory-reared Culicoides nubeculosus (Meigen) under fluctuating temperatures (16 and 28 °C) and with access to water or water and blood was on average 6.4 days and 8.9 days, respectively, which was around one third of the lifespan of siblings with access to sugar or sugar and blood (22.2 days and 27.1 days, respectively). Access to honeydew significantly increased the midge's longevity, whereas the provision of extrafloral nectaries had no impact. Females with access to sugar produced a significantly higher number of eggs (65.5 ± 5.2) than their starved sisters (45.4 ± 8.4). More than 80% of field-caught female Culicoides from the two most abundant European groups, Obsoletus (n = 2243) and Pulicaris (n = 805), were fructose-positive. Fructose-positivity was high in all physiological stages and no seasonal variability was noted. The high rate of natural sugar feeding of Culicoides offers opportunities for the development of novel control strategies using toxic sugar baits and for the monitoring of vector-borne diseases using sugar-treated FTA (nucleic acid preservation) cards in the field.


An Aeromonas caviae genomic island is required for both O-antigen lipopolysaccharide biosynthesis and flagellin glycosylation.
Aeromonas caviae Sch3N possesses a small genomic island that is involved in both flagellin glycosylation and lipopolysaccharide (LPS) O-antigen biosynthesis. This island appears to have been laterally acquired as it is flanked by insertion element-like sequences and has a much lower G+C content than the average aeromonad G+C content. Most of the gene products encoded by the island are orthologues of proteins that have been shown to be involved in pseudaminic acid biosynthesis and flagellin glycosylation in both Campylobacter jejuni and Helicobacter pylori. Two of the genes, lst and lsg, are LPS specific as mutation of them results in the loss of only a band for the LPS O-antigen. Lsg encodes a putative Wzx flippase, and mutation of Lsg affects only LPS; this finding supports the notion that flagellin glycosylation occurs within the cell before the flagellins are exported and assembled and not at the surface once the sugar has been exported. The proteins encoded by flmA, flmB, neuA, flmD, and neuB are thought to make up a pseudaminic acid biosynthetic pathway, and mutation of any of these genes resulted in the loss of motility, flagellar expression, and a band for the LPS O-antigen. Furthermore, pseudaminic acid was shown to be present on both flagellin subunits that make up the polar flagellum filament, to be present in the LPS O-antigen of the A. caviae wild-type strain, and to be absent from the A. caviae flmD mutant strain.


[Study on pathogenetic role of myeloperoxidase, tumor necrosis factor alpha, interferon gamma in chronic airway infection with Pseudomonas aeruginosa].
This study was performed to demonstrate the pathogenetic role of mucoid-alginate produced by Pseudomonas aeruginosa in relation to immuno-interaction in host cells. The obtained results are as follows. 1. Clinical Observation The concentration of MPO, TNF alpha, and IFN gamma in peripheral blood of 37 cases of chronic airway infection were observed. The cases were divided into the groups with mucoid strain positive (21 cases) and negative (16 cases), and each group was also divided into subgroups with active clinical symptom and without. High concentration of serum IFN gamma was shown in the group of mucoid strain positive cases (0.10 +/- 0.08 IU/ml in healthy volunteers group, 0.54 +/- 0.56 IU/ml in mucoid group with symptom: p < 0.01, 0.09 +/- 0.15 IU/ml in mucoid group without symptom, 0.13 +/- 0.13 IU/ml in non-mucoid group with symptom, 0.09 +/- 0.13 IU/ml in non-mucoid group without symptom). But plasma concentrations of MPO and serum concentrations of TNF alpha were not increased in all groups, except MPO in non-mucoid group with symptom. It indicates that IFN gamma has a characteristic role in chronic airway infection with mucoid Pseudomonas aeruginosa (P. aeruginosa). 2. Experimental Observation One hundred twenty-two of alginate-immunized mice were made by 5 times of intraabdominal injection with purified alginate extracted from mucoid P. aeruginosa PT-1252. 4 x 10(6) CFU/body of mucoid P. aeruginosa PT-1252 or 20 micrograms/body of purified alginate was intubated per trachea into the immunized mice, and changes in mean fluorescence intensity (MFI) of MPO and TNF alpha on neutrophils and IFN gamma on CD3 positive lymphocytes in BALF were observed. MFI of MPO (99.7 +/- 16.5-->177.0 +/- 23.1: p < 0.001) and TNF alpha (59.2 +/- 13.3-->197.0 +/- 62.0: p < 0.001) were increased on day 2 and they were kept up till on day 30 in P. aeruginosa group. But in alginate group they were temporally decreased on day 2, and they were gradually increased (MPO: 99.7 +/- 16.5-->212.6 +/- 12.1: p < 0.001, TNF alpha: 59.2 +/- 13.3-->162.4 +/- 30.9: p < 0.01). The temporally decrease of MPO and TNF alpha on day 2 in alginate group may suggest that a large amount of alginate inhibits neutrophils chemotaxis. MFI of IFN gamma in P. aeruginosa was also increased (110.0 +/- 32.9-->198.3 +/- 23.0: p < 0.01) on day 2 and it was gradually decreased on day 30 (156.0 +/- 13.8). In alginate group, MFI of IFN gamma in BALF was increased on day 5 (110.0 +/- 32.9-->201.0 +/- 49.3: p < 0.001) and kept a high level till day 30 (223.0 +/- 57.5: p < 0.01). In morphological, the moderate grade of lymphocytes infiltration observed in intersitial area of lung tissue on both groups of P. aeruginosa and alginate intubation mice. The findings was continuously seen from day 5 through day 30. The lymphocytes infiltration appeared in lung tissue can be though as an result of immuno-interaction with IFN gamma positive lymphocytes and mucoid-alginate. Inferences are drawn from the clinical results and the experimental ones that the CD3 positive IFN gamma lymphocytes expressed by immunological interaction with mucoid-alginate and the IFN gamma lymphocytes (Th1 like cells) plays an important role on advancement of chronic airway infection with mucoid P. aeruginosa.


A tale of two toxicities: malformed selenoproteins and oxidative stress both contribute to selenium stress in plants.
Despite selenium's toxicity in plants at higher levels, crops supply most of the essential dietary selenium in humans. In plants, inorganic selenium can be assimilated into selenocysteine, which can replace cysteine in proteins. Selenium toxicity in plants has been attributed to the formation of non-specific selenoproteins. However, this paradigm can be challenged now that there is increasingly abundant evidence suggesting that selenium-induced oxidative stress also contributes to toxicity in plants. This Botanical Briefing summarizes the evidence indicating that selenium toxicity in plants is attributable to both the accumulation of non-specific selenoproteins and selenium-induced oxidative stress. Evidence is also presented to substantiate the claim that inadvertent selenocysteine replacement probably impairs or misfolds proteins, which supports the malformed selenoprotein hypothesis. The possible physiological ramifications of selenoproteins and selenium-induced oxidative stress are discussed. Malformed selenoproteins and oxidative stress are two distinct types of stress that drive selenium toxicity in plants and could impact cellular processes in plants that have yet to be thoroughly explored. Although challenging, deciphering whether the extent of selenium toxicity in plants is imparted by selenoproteins or oxidative stress could be helpful in the development of crops with fortified levels of selenium.