[An adolescent with lumbosacral instability due to the presence of a hydatid cyst: a case report].
Hydatidosis, caused by Echinococcus granulosus, affects the spinal column in 0.5-2% of all cases, with the sacral column being a rare location typically observed only in adults. The case is reported of a 14-year-old boy from Juliaca (Puno, Peru) who presented with chronic lower back pain and cauda equina syndrome. He was initially attended to in Juliaca, and then transferred to the Hospital Edgardo Rebagliati Martins (Lima, Peru) with a diagnosis of sacral tumor. In magnetic resonance imaging, a multicystic tumor was seen with L5 and S1 lysis and instability. The surgical treatment was an L5- S2 laminectomy and extraction of multiple hydatid cysts followed by lumbopelvic stabilization with transpedicular screws. Post-operatively, he was prescribed albendazol (15 mg/kg/d) for six months. Hydatid cysts should be considered within the differential diagnosis when there are cystic lesions in the spinal column, particularly in endemic areas such as Peru.


Genomic DNA extraction optimization and validation for genome sequencing using the marine gastropod Kellet's whelk.
Next-generation sequencing technologies, such as Nanopore MinION, Illumina Hiseq and Novaseq, and PacBio Sequel II, hold immense potential for advancing genomic research on non-model organisms, including the vast majority of marine species. However, application of these technologies to marine invertebrate species is often impeded by challenges in extracting and purifying their genomic DNA due to high polysaccharide content and other secondary metabolites. In this study, we help resolve this issue by developing and testing DNA extraction protocols for Kellet's whelk (Kelletia kelletii), a subtidal gastropod with ecological and commercial importance, by comparing four DNA extraction methods commonly used in marine invertebrate studies. In our comparison of extraction methods, the Salting Out protocol was the least expensive, produced the highest DNA yields, produced consistent high DNA quality, and had low toxicity. We validated the protocol using an independent set of tissue samples, then applied it to extract high-molecular-weight (HMW) DNA from over three thousand Kellet's whelk tissue samples. The protocol demonstrated scalability and, with added clean-up, suitability for RAD-seq, GT-seq, as well as whole genome sequencing using both long read (ONT MinION) and short read (Illumina NovaSeq) sequencing platforms. Our findings offer a robust and versatile DNA extraction and clean-up protocol for supporting genomic research on non-model marine organisms, to help mediate the under-representation of invertebrates in genomic studies.


Differential regulation of DNA synthesis by nitric oxide and hydroxyurea in vascular smooth muscle cells.
We investigated the influence of nitrovasodilators on DNA synthesis in cultured human aortic smooth muscle cells and explored the hypothesis that nitric oxide (NO) is directly involved in mediating the inhibitory effects of hydroxyurea on DNA synthesis. Both NO and hydroxyurea inhibited ongoing DNA synthesis and S phase progression in our cells. Exogenous deoxynucleosides partially reversed this inhibition, suggesting that ribonucleotide reductase is a primary target for both NO and hydroxyurea. Nitrovasodilators inhibited DNA synthesis by releasing NO, as detected by chemiluminescence and as shown by the reversal of DNA synthesis inhibition by NO scavengers. This inhibition appears to occur via a cGMP-independent mechanism. In contrast, hydroxyurea did not produce a detectable NO signal, and NO scavengers had no influence on its inhibition of DNA synthesis, suggesting that NO does not mediate the inhibitory action of hydroxyurea in our system. Furthermore, the action of nitrovasodilators and hydroxyurea on DNA synthesis differed according to redox sensitivity. The redox agents N-acetyl-L-cysteine and ascorbate reversed NO inhibition of DNA synthesis and had no effect on DNA synthesis inhibition caused by hydroxyurea.


Event-related potentials and oscillatory brain responses associated with semantic and Stroop-like interference effects in overt naming.
Picture-word interference is a widely employed paradigm to investigate lexical access in word production: Speakers name pictures while trying to ignore superimposed distractor words. The distractor can be congruent to the picture (pictured cat, word cat), categorically related (pictured cat, word dog), or unrelated (pictured cat, word pen). Categorically related distractors slow down picture naming relative to unrelated distractors, the so-called semantic interference. Categorically related distractors slow down picture naming relative to congruent distractors, analogous to findings in the colour-word Stroop task. The locus of semantic interference and Stroop-like effects in naming performance has recently become a topic of debate. Whereas some researchers argue for a pre-lexical locus of semantic interference and a lexical locus of Stroop-like effects, others localise both effects at the lexical selection stage. We investigated the time course of semantic and Stroop-like interference effects in overt picture naming by means of event-related potentials (ERP) and time-frequency analyses. Moreover, we employed cluster-based permutation for statistical analyses. Naming latencies showed semantic and Stroop-like interference effects. The ERP waveforms for congruent stimuli started diverging statistically from categorically related stimuli around 250 ms. Deflections for the categorically related condition were more negative-going than for the congruent condition (the Stroop-like effect). The time-frequency analysis revealed a power increase in the beta band (12-30 Hz) for categorically related relative to unrelated stimuli roughly between 250 and 370 ms (the semantic effect). The common time window of these effects suggests that both semantic interference and Stroop-like effects emerged during lexical selection.


[Clinical-epidemiologic profile of Dengue suspicious cases in a district in the south zone of Teresina, PI, Brazil].
The objective of the study was to establish a clinical-epidemiologic profile of Dengue suspicious cases in a district in the south zone of Teresina, PI, Brazil. This study is a descriptive-exploratory survey in which it was applied a questionnaire to 28 residents of the mentioned district. 88.9% had garbage collection 3 times per week; 57.1% of the deposits had construction material or car parts; 75% destine served waters to opencast; 60.7% had developed first-infection, 75% were submitted to serology, 33.3% cases were confirmed. More effectiveness of sanitary action by government and population involvement in the development of preventive programs is missed in the fight against Dengue.


Prostate cancer treatment ascertained from several sources: analysis of disagreement and error.
Treatment data for prostate cancer can be obtained from a variety of sources. Each of these sources has its own strengths and weaknesses and is subject to error. In a population-based cohort of 319 prostate cancer patients, data on treatment were obtained from five sources: two patient interviews at 6 and 12 months after diagnosis, primary caregiver interviews, physician questionnaires, and medical records. Inter-reporting agreement and accuracy of reporting (compared with medical records) were assessed. Multivariate analyses examined patient, caregiver, and physician characteristics as determinants of reporting error. The agreement among different reporting methods was generally good to excellent for prostatectomy and brachytherapy (kappa range 0.70-0.90) and fair to good (kappa range 0.35-0.75) for external beam radiation and hormonal treatment. Compared with medical records, the interview- and questionnaire-based data collection methods were more accurate for prostatectomy and brachytherapy than for external beam radiation and hormonal therapy. Using medical records as the 'gold standard', patient and caregiver interviews at 6 months after the diagnosis had higher sensitivity and specificity than other reporting sources. Interviews of prostate cancer patients and caregivers are useful alternatives to medical record abstraction, particularly if carried out during, or soon after, treatment.


Centralization of histocompatibility laboratories: impact on organ allocation efficiency and outcomes of cadaveric renal transplantation.
This project was undertaken to determine whether centralization of histocompatibility laboratory services for renal transplants performed within eastern Pennsylvania could improve the efficiency of allograft allocation and short-term allograft function. A nonconcurrent cohort study was performed comparing renal allografts transplanted between September 15, 1993, and September 14, 1994, to those transplanted between September 15, 1994, and September 14, 1995. All allografts were procured and allocated by the Delaware Valley Transplant Program, the organ procurement agency in eastern Pennsylvania. Cold preservation time and delayed allograft function were used to measure efficiency of allograft allocation and short term allograft function, respectively. The mean cold preservation time was reduced from 25.08 hours to 20.68 hours (P < 0.001). The percentage of delayed allograft function was 19.9 and 17.4 for the pre- and postcentralization groups, respectively (P = 0.5). Therefore, centralization of histocompatibility tissue typing was a regionally effective process intervention for reducing cold preservation time without adversely impacting short-term graft function. The magnitude of this reduction varied between individual centers. Further investigation is required to determine the effect on long-term allograft function and system wide costs.


Intracerebral hemorrhage after intravenous t-PA therapy for ischemic stroke. The NINDS t-PA Stroke Study Group.
We sought to identify variables associated with intracerebral hemorrhage in patients with acute ischemic stroke who receive tissue plasminogen activator (t-PA). We performed subgroup analyses of data from a randomized, double-blind, placebo-controlled trial of intravenous t-PA administered to stroke patients within 3 hours of onset. Using multivariable regression modeling procedures, we assessed the relationship of baseline and after-treatment variables with symptomatic and asymptomatic intracerebral hemorrhage during the first 36 hours after treatment. Overall, t-PA-treated patients had an increase in the absolute risk of symptomatic intracerebral hemorrhage of 6% and a decrease in the absolute risk of 3-month mortality of 4% compared with placebo-treated patients. The only variables independently associated with an increased risk of symptomatic intracerebral hemorrhage in the final multivariable logistic regression model for the 312 t-PA-treated patients were the severity of neurological deficit as measured by the National Institutes of Health Stroke Scale score (five categories; odds ratio [OR], 1.8; 95% confidence interval [CI], 1.2 to 2.9) and brain edema (defined as acute hypodensity) or mass effect by CT before treatment (OR, 7.8; 95% CI, 2.2 to 27.1). This final model correctly predicted those t-PA-treated patients who would or would not have a symptomatic hemorrhage with only 57% efficiency. In the subgroup of patients with a severe neurological deficit, t-PA-treated patients were more likely than placebo-treated patients to have a favorable 3-month outcome (adjusted OR based on multiple outcomes, 4.3; 95% CI, 1.6 to 11.9). These results were similar for the subgroup with edema or mass effect by CT (adjusted OR, 3.4; 95% CI, 0.6 to 20.7). The likelihood of severe disability or death was similar for t-PA- and placebo-treated patients with these two baseline characteristics. Despite a higher rate of intracerebral hemorrhage, patients with severe strokes or edema or mass effect on the baseline-CT are reasonable candidates for t-PA, if it is administered within 3 hours of onset.


Hybrid silencers with micro-perforated panels and internal partitions.
A sub-structuring approach, along with a unit cell treatment, is proposed to model expansion chamber silencers with internal partitions and micro-perforated panels (MPPs) in the absence of internal flow. The side-branch of the silencer is treated as a combination of unit cells connected in series. It is shown that, by connecting multiple unit cells with varying parameters, the noise attenuation bandwidth can be enlarged. With MPPs, the hybrid noise attenuation mechanism of the silencer is revealed. Depending on the size of the perforation hole, noise attenuation can be dominated by dissipative, reactive, or combined effects together. For a broadband sound absorption, the hole size, together with the perforation ratio and other parameters, can be optimized to strike a balance between the dissipative and reactive effect, for ultimately achieving the desired noise attenuation performance within a prescribed frequency region. The modular nature of the proposed formulation allows doing this in a flexible, accurate, and cost effective manner. The accuracy of the proposed approach is validated through comparisons with finite element method and experiments.


Motor proficiency of 6- to 9-year-old children with speech and language problems.
This study compared the gross motor skills of school-age children (mean age 7 y 8 mo, range 6-9 y) with developmental speech and language disorders (DSLDs; n=105; 76 males, 29 females) and typically developing children (n=105; 76 males, 29 females). The relationship between the performance parameters and the children's age was investigated as well as the role of the type of DSLD. The children with DSLDs were classified by their schools' speech and language therapists into three subgroups: children with speech disorders (n=16), those with language disorders (n=41), or those with both (n=48). They were tested with the Test of Gross Motor Development, 2nd edition. Compared with their typically developing peers, all three DSLD subgroups scored lower on the locomotor (all p values<0.001) and object control sub tests (all p values<0.001). Significant performance differences were found between the three types of DSLD (all p values<0.01) where the children with language disorders only performed better. Older children performed better than the younger ones (plocomotor=0.029, pobject control<0.001), but the magnitude of differences between the children with DSLDs and their peers did not change with increasing age. Children with DSLDs have poor gross motor skills. Although the performance of children with DSLDs improves with increasing age, it lags behind that of typically developing children. The present results emphasize the importance of early diagnosis and intervention for children with motor deficits.


Chronic and acute melatonin effects in gerbil global forebrain ischemia: long-term neural and behavioral outcome.
Melatonin attenuates the short-term consequences of brain ischemia in several animal models. However, there is scant information regarding its efficacy for improving the long-term outcome. To further address that issue, we subjected gerbils to 5-min bilateral carotid occlusion. Some gerbils received acute peri-surgical administration of melatonin while others received continuous melatonin in their water. The gerbils' brains were histologically assessed at 20 wk postsurgery. Chronic but not acute melatonin attenuated ischemia-induced hyperactivity at 3 days postsurgery. Twenty weeks postsurgery, the ischemic gerbils showed varying degrees of bilateral loss of hippocampal CA1 pyramidal cells and elevation of glial fibrillary acidic protein immunoreactivity there. Both the cell loss and the immunoreactivity were markedly asymmetrical for some gerbils. Neither acute nor chronic melatonin altered this pattern of CA1 cell loss and glial immunoreactivity increase. Ischemia increased the number of CA1 cells that were immunoreactive for doublecortin (DCX), a marker for newborn neurons. This increase in CA1 DCX expression was not affected by either melatonin treatment. However, both acute and chronic melatonin reduced the number of DCX immunoreactive neurons in the dentate gyrus. Thus, neither acute nor chronic melatonin altered the long-term neural outcome of forebrain ischemia, although chronic administration seemed to attenuate the short-term behavioral effect. It is suggested that persistently high brain levels of melatonin may be essential for long-term neuroprotection against ischemia. The possibility that melatonin may modulate hippocampal neurogenesis merits further exploration both in normal animals and in models of brain insult.


Safety of pregnancy after breast cancer in young women with hormone receptor-positive disease: a systematic review and meta-analysis.
Despite increasing evidence on the safety of pregnancy after anticancer treatments in breast cancer survivors, many physicians and patients remain concerned about a potential risk of pregnancy specifically in the case of hormone receptor-positive breast cancer. A systematic literature search of Medline, Embase and Cochrane library with no language or date restriction up to 31 March 2023 was carried out. To be included, articles had to be retrospective and prospective case-control and cohort studies as well as clinical trials comparing survival outcomes of premenopausal women with or without a pregnancy after prior diagnosis of hormone receptor-positive breast cancer. Disease-free survival (DFS) and overall survival (OS) were the outcomes of interest. Pooled hazard ratios (HRs) with 95% confidence intervals (CIs) were calculated. Study protocol is registered in PROSPERO (n. CRD42023394232). Out of 7796 screened studies, 8 were eligible to be included in the final analysis. A total of 3805 patients with hormone receptor-positive invasive early breast cancer were included in these studies, of whom 1285 had a pregnancy after breast cancer diagnosis. Median follow-up time ranged from 3.8 to 15.8 years and was similar in the pregnancy and non-pregnancy cohorts. In three studies (n = 987 patients) reporting on DFS, no difference was observed between patients with and those without a subsequent pregnancy (HR 0.96, 95% CI 0.75-1.24, P = 0.781). In the six studies (n = 3504 patients) reporting on OS, patients with a pregnancy after breast cancer had a statistically significant better OS than those without a pregnancy (HR 0.46, 95% CI 0.27-0.77, P < 0.05). This systematic review and meta-analysis of retrospective cohort studies provides updated evidence that having a pregnancy in patients with prior history of hormone receptor-positive invasive early breast cancer appears safe without detrimental effect on prognosis.


OSNN: An Online Semisupervised Neural Network for Nonstationary Data Streams.
Learning from data streams that emerge from nonstationary environments has many real-world applications and poses various challenges. A key characteristic of such a task is the varying nature of target functions and data distributions over time (concept drifts). Most existing work relies solely on labeled data to adapt to concept drifts in classification problems. However, labeling all instances in a potentially life-long data stream is frequently prohibitively expensive, hindering such approaches. Therefore, we propose a novel algorithm to exploit unlabeled instances, which are typically plentiful and easily obtained. The algorithm is an online semisupervised radial basis function neural network (OSNN) with manifold-based training to exploit unlabeled data while tackling concept drifts in classification problems. OSNN employs a novel semisupervised learning vector quantization (SLVQ) to train network centers and learn meaningful data representations that change over time. It uses manifold learning on dynamic graphs to adjust the network weights. Our experiments confirm that OSNN can effectively use unlabeled data to elucidate underlying structures of data streams while its dynamic topology learning provides robustness to concept drifts.


TGF-beta signaling may play a role in the development of goblet cell hyperplasia in a mouse model of allergic rhinitis.
Transforming growth factor-beta (TGF-beta) levels are elevated in the nasal mucosa in allergic rhinitis. However, because TGF-beta is secreted extracellulary in latent complexes, it remains unclear whether the local TGF-beta expression actually drives active signaling and affects the pathophysiology of allergic rhinitis. The objective of this study is to investigate whether TGF-beta signaling is activated in allergic rhinitis and plays a role in the pathophysiology of allergic rhinitis. An ovabumin (OVA)-sensitized and -nasally challenged mouse model of allergic rhinitis was established and phosphorylation of Smad2 in the nasal mucosa was examined by immunohistochemistry. In addition, the effects of the pharmacological inhibition of endogenous TGF-beta signaling on the allergic rhinitis model were histologically examined. Furthermore, phosphorylation of Smad2 in the nasal mucosa samples obtained from patients with allergic rhinitis was also evaluated. In the mouse model of allergic rhinitis, OVA challenge induced phosphorylation of Smad2 predominantly in epithelial cells in the nasal mucosa. In addition, the administration of an inhibitor of TGF-beta type I receptor kinase activity during OVA challenge suppressed goblet cell hyperplasia in the nasal mucosa. Furthermore, phosphorylated Smad2 expression increased in nasal epithelial cells in patients with allergic rhinitis. These results suggest that TGF-beta signaling is activated in epithelial cells in the nasal mucosa in allergic rhinitis and may contribute to the development of goblet cell hyperplasia.


confFuse: High-Confidence Fusion Gene Detection across Tumor Entities.
Background: Fusion genes play an important role in the tumorigenesis of many cancers. Next-generation sequencing (NGS) technologies have been successfully applied in fusion gene detection for the last several years, and a number of NGS-based tools have been developed for identifying fusion genes during this period. Most fusion gene detection tools based on RNA-seq data report a large number of candidates (mostly false positives), making it hard to prioritize candidates for experimental validation and further analysis. Selection of reliable fusion genes for downstream analysis becomes very important in cancer research. We therefore developed confFuse, a scoring algorithm to reliably select high-confidence fusion genes which are likely to be biologically relevant. Results: confFuse takes multiple parameters into account in order to assign each fusion candidate a confidence score, of which score ≥8 indicates high-confidence fusion gene predictions. These parameters were manually curated based on our experience and on certain structural motifs of fusion genes. Compared with alternative tools, based on 96 published RNA-seq samples from different tumor entities, our method can significantly reduce the number of fusion candidates (301 high-confidence from 8,083 total predicted fusion genes) and keep high detection accuracy (recovery rate 85.7%). Validation of 18 novel, high-confidence fusions detected in three breast tumor samples resulted in a 100% validation rate. Conclusions: confFuse is a novel downstream filtering method that allows selection of highly reliable fusion gene candidates for further downstream analysis and experimental validations. confFuse is available at https://github.com/Zhiqin-HUANG/confFuse.


Use of laboratory data to reduce the time taken to detect new diseases: VIDA to FarmFile.
The analysis of laboratory data can provide information about the health of livestock populations; in Great Britain the Veterinary Investigation Diagnosis Analysis (VIDA) system has provided such data since 1975. However VIDA covers only known diagnoses, with limited epidemiological characterisation. The unexpected outbreak of bse showed that it was necessary to improve surveillance to detect new diseases, and a necessary update of the VIDA database for the millennium date change provided the opportunity. The information required to enhance the value of laboratory data was identified, a new form and database, 'FarmFile', were designed to record it, and they began to be used in 1999. The detection of new diseases depends on making comparisons with the expected or 'usual' levels of unexplained disease. The data are analysed quarterly to assess any changes in the levels of unexplained disease in different species, categorised in terms of clinical sign or body system, by comparison with previous years. No new diseases have been detected either through FarmFile or more traditional means since the new analyses started in earnest in 2004, but they have indicated that an unexplained event was not a new disease of concern, and developments continue to improve the system's sensitivity and specificity.


Epidermal growth factor-induced heparanase nucleolar localization augments DNA topoisomerase I activity in brain metastatic breast cancer.
Identification of molecular mechanisms responsible for brain metastatic breast cancer (BMBC) is imperative to develop novel therapies. However, current understanding of the molecular circuitry that governs BMBC dissemination remains fragmentary. Heparanase (HPSE) is the only functional mammalian endoglycosidase whose activity correlates with cancer metastasis, angiogenesis, and the reduced postoperative survival of cancer patients, making it an active target for anticancer therapeutics. We hypothesized that human epidermal growth factor receptor 2 (HER2)/epidermal growth factor receptor (EGFR) activation promotes HPSE function in human BMBC. To address this, we examined HPSE content, activity, and intracellular trafficking in a HER2/EGFR-expressing BMBC model system and show that HPSE is present, functional, and correlates with HER2 status. Further, we showed that EGF induced nucleolar translocation of HPSE in these cells in a dose- and time-dependent manner upon activation of HER2/EGFR. Knockdowns of HER2/EGFR by small interference RNA abolished EGF-induced HPSE nucleolar translocalization. It was also noted that nucleolar HPSE modulates DNA topoisomerase I (Topo I), an enzyme that is highly present in nucleoli, essential for DNA replication and transcription in a variety of tumors, and inhibited by heparan sulfate. Evidence is provided that HPSE can regulate Topo I activity, which subsequently affects BMBC cell proliferation. Finally, we showed that the nucleolar presence of HPSE with Topo I colocalization is detected only in HER2-overexpressing BMBC patient specimens. Altogether, these findings support the notion that HPSE is a critical downstream target of HER2 mechanisms driving BMBC and is potentially relevant for BMBC therapeutic interventions.


Medical education is a growing international field of research. However, no overview exists of Norwegian researchers' contribution to the field. This study aimed to identify and describe medical education research originating from Norwegian research institutions. We performed targeted searches in Web of Science, PubMed, ProQuest Education and Cristin, and identified published research within the field from Norwegian institutions. Using bibliometric analyses, we mapped patterns in research output over time, research communities that have published their work, and which publication channels were used. We also categorized the research articles according to methodology, empirical origin, educational level and topic. We found 423 published research articles linked to Norwegian research institutions and an increase in the number of publications in the last decade. The studies were published in 141 different scientific journals. Of these, the Journal of the Norwegian Medical Association had 94 articles (22 %). A total of 122 (29 %) articles were published in key medical education journals. Altogether 249 (59 %) of the studies were quantitative and 262 (62 %) were based on empirical data from Norway. A little more than half, 237 (56 %), focused on undergraduate medical education. Researchers affiliated to the University of Oslo were involved in 173 (43 %) articles. The most frequently researched topics were curricula, teaching and learning technology, retention and career choices. Research activity within the field of medical education is increasing in Norway, but few Norwegian researchers publish in the specialised medical education journals.


Longevity effect of a polysaccharide from Chlorophytum borivilianum on Caenorhabditis elegans and Saccharomyces cerevisiae.
The traditional Indian medicine, Ayurveda, provides insights and practical solutions towards a healthy life style. Rasayana is a branch of Ayurveda known for preserving and promoting health, enhancing the quality of life and delaying the aging process. In the traditional knowledge, the Rasayana herb, Chlorophytum borivilianum (C. borivilanum) is regarded as a general health promoting tonic that delays aging and increases lifespan, cognitive function and physical strength. Aging is a complex and multifactorial physiological phenomenon that manifests itself over a wide range of biological systems, tissues, and functions. Longevity is an obvious marker of physiological aging. Simple model systems such as the single-cell budding yeast Saccharomyces cerevisiae (S. cerevisiae) and the nematode, Caenorhabditis elegans (C. elegans) are widely used to study the aging process and longevity. Here, we show that a polysaccharide fraction obtained from C. borivilianum increases the lifespan of S. cerevisiae and C. elegans, using an automated screening platform (ChronoscreenTM). Chemical analysis of this extract revealed a low molecular weight polysaccharide of 1000 Da, predominantly comprising Glu1→6Glu linkage. This polysaccharide showed significant dose-dependent extension of the median lifespan of S. cerevisiae by up to 41% and of the median lifespan of C. elegans by up to 10%. Taking cue from these results and the traditionally described benefits of Rasayanas on skin rejuvenation, we tested in vitro the polysaccharide for potential skin benefits. In a keratinocyte culture, we observed that this polysaccharide increased cell proliferation significantly, and induced synthesis of hyaluronic acid (HA), a well-known extracellular matrix component. Furthermore, when added to culture medium of human reconstructed epidermis, we observed an enhanced production of epidermal markers, e.g. CD44 and HA that are otherwise diminished in aged skin. Together, these results suggest that in addition to life-span extension of S. cerevisiae and C. elegans, a polysaccharide from the Rasayana herb, C. borivilianum may have beneficial effects on skin aging parameters.


Health problems and disability in long-term sickness absence: ICF coding of medical certificates.
The purpose of this study was to test the feasibility of International Classification of Functioning, Disability and Health (ICF) and to explore the distribution, including gender differences, of health problems and disabilities as reflected in long-term sickness absence certificates. A total of 433 patients with long sick-listing periods, 267 women and 166 men, were included in the study. All certificates exceeding 28 days of sick-listing sent to the local office of the Swedish Social Insurance Administration of a municipality in the Stockholm area were collected during four weeks in 2004-2005. ICD-10 medical diagnosis codes in the certificates were retrieved and free text information on disabilities in body function, body structure or activity and participation were coded according to ICF short version. In 89.8% of the certificates there were descriptions of disabilities that readily could be classified according to ICF. In a reliability test 123/131 (94%) items of randomly chosen free text information were identically classified by two of the authors. On average 2.4 disability categories (range 0-9) were found per patient; the most frequent were 'Sensation of pain' (35.1% of the patients), 'Emotional functions' (34.1%), 'Energy and drive functions' (22.4%), and 'Sleep functions' (16.9%). The dominating ICD-10 diagnostic groups were 'Mental and behavioural disorders' (34.4%) and 'Diseases of the musculoskeletal system and connective tissue' (32.8%). 'Reaction to severe stress and adjustment disorders' (14.7%), and 'Depressive episode' (11.5%) were the most frequent diagnostic codes. Disabilities in mental functions and activity/participation were more commonly described among women, while disabilities related to the musculoskeletal system were more frequent among men. Both ICD-10 diagnoses and ICF categories were dominated by mental and musculoskeletal health problems, but there seems to be gender differences, and ICF classification as a complement to ICD-10 could provide a better understanding of the consequences of diseases and how individual patients can cope with their health problems. ICF is feasible for secondary classifying of free text descriptions of disabilities stated in sick-leave certificates and seems to be useful as a complement to ICD-10 for sick-listing management and research.


Oleoylethanolamine and palmitoylethanolamine modulate intestinal permeability in vitro via TRPV1 and PPARα.
Cannabinoids modulate intestinal permeability through cannabinoid receptor 1 (CB1). The endocannabinoid-like compounds oleoylethanolamine (OEA) and palmitoylethanolamine (PEA) play an important role in digestive regulation, and we hypothesized they would also modulate intestinal permeability. Transepithelial electrical resistance (TEER) was measured in human Caco-2 cells to assess permeability after application of OEA and PEA and relevant antagonists. Cells treated with OEA and PEA were stained for cytoskeletal F-actin changes and lysed for immunoassay. OEA and PEA were measured by liquid chromatography-tandem mass spectrometry. OEA (applied apically, logEC50 -5.4) and PEA (basolaterally, logEC50 -4.9; apically logEC50 -5.3) increased Caco-2 resistance by 20-30% via transient receptor potential vanilloid (TRPV)-1 and peroxisome proliferator-activated receptor (PPAR)-α. Preventing their degradation (by inhibiting fatty acid amide hydrolase) enhanced the effects of OEA and PEA. OEA and PEA induced cytoskeletal changes and activated focal adhesion kinase and ERKs 1/2, and decreased Src kinases and aquaporins 3 and 4. In Caco-2 cells treated with IFNγ and TNFα, OEA (via TRPV1) and PEA (via PPARα) prevented or reversed the cytokine-induced increased permeability compared to vehicle (0.1% ethanol). PEA (basolateral) also reversed increased permeability when added 48 or 72 h after cytokines (P < 0.001, via PPARα). Cellular and secreted levels of OEA and PEA (P < 0.001-0.001) were increased in response to inflammatory mediators. OEA and PEA have endogenous roles and potential therapeutic applications in conditions of intestinal hyperpermeability and inflammation.-Karwad, M. A., Macpherson, T., Wang, B., Theophilidou, E., Sarmad, S., Barrett, D. A., Larvin, M., Wright, K. L., Lund, J. N., O'Sullivan, S. E. Oleoylethanolamine and palmitoylethanolamine modulate intestinal permeability in vitro via TRPV1 and PPARα.


Immune Microenvironment Landscape in CNS Tumors and Role in Responses to Immunotherapy.
Despite the important evolution of immunotherapeutic agents, brain tumors remain, in general, refractory to immune therapeutics. Recent discoveries have revealed that the glioma microenvironment includes a wide variety of immune cells in various states that play an important role in the process of tumorigenesis. Anti-tumor immune activity may be occurring or induced in immunogenic hot spots or at the invasive edge of central nervous system (CNS) tumors. Understanding the complex heterogeneity of the immune microenvironment in gliomas will likely be the key to unlocking the full potential of immunotherapeutic strategies. An essential consideration will be the induction of immunological effector responses in the setting of the numerous aspects of immunosuppression and evasion. As such, immune therapeutic combinations are a fundamental objective for clinical studies in gliomas. Through immune profiling conducted on immune competent murine models of glioma and ex vivo human glioma tissue, we will discuss how the frequency, distribution of immune cells within the microenvironment, and immune modulatory processes, may be therapeutically modulated to lead to clinical benefits.


Targeting CXCR4 reverts the suppressive activity of T-regulatory cells in renal cancer.
With the intent to identify biomarkers in renal cell carcinoma (RCC) the functional status of T-regulatory cells (Tregs) was investigated in primary RCC. Tregs were isolated from tumoral-(TT), peritumoral tissue-(PT) and peripheral blood-(PB) of 42 primary RCC patients and function evaluated through effector T cells (Teff) proliferation, cytokines release and demethylation of Treg Specific Region (TSDR). The highest value of Tregs was detected in TT with the uppermost amount of effector-Tregs-(CD4+CD25hiFOXP3hiCD45RA-). PB-RCC Tregs efficiently suppress Teff proliferation compared to healthy donor (HD)-Tregs and, at the intrapatient evaluation, TT-derived Tregs were the most suppressive. Higher demethylation TSDR was detected in TT- and PB-RCC Tregs vs HD-Tregs (P <0,001). CXCR4 is highly expressed on Tregs, thus we wished to modulate Tregs function through CXCR4 inhibition. CXCR4 antagonism, elicited by a new peptidic antagonist, Peptide-R29, efficiently reversed Tregs suppression of Teff proliferation. Thus Tregs functional evaluation precisely reflects Tregs status and may be a reliable biomarker of tumoral immune response. In addition, treatment with CXCR4 antagonist, impairing Tregs function, could improve the anticancer immune response, in combination with conventional therapy and/or immunotherapy such as checkpoints inhibitors.


Highly branched platinum nanolance assemblies by polyallylamine functionalization as superior active, stable, and alcohol-tolerant oxygen reduction electrocatalysts.
The chemical functionalization of platinum (Pt) nanostructures is becoming a new trend in electrocatalysts designs. Meanwhile, highly branched Pt nanostructures are highly exciting electrocatalysts with high activity and stability owing to their specific physical and chemical properties. In this work, the polyallylamine (PAH) functionalized Pt nanolance assemblies (Pt NLAs) have been successfully synthesized by chemical reduction of PAH-Pt(II) complex using formaldehyde (HCHO) in a two-phase water-complex system. The as-prepared Pt NLAs are highly branched and three-dimensionally (3D) interconnected nanostructures, which are composed of many long Pt nanolances in various directions. PAH functionalization improves the electrocatalytic activity of the Pt NLAs for an oxygen reduction reaction (ORR) because of high interface proton concentration on the Pt surface and excellent anti-oxidation ability of the Pt nanolances. Meanwhile, the PAH molecules bound on the Pt NLAs surface act as barrier networks to restrain accessibility of alcohol, exhibiting a high ORR selectivity. In addition, the PAH functionalized Pt NLAs show excellent durability for the ORR due to their particular 3D interconnected structure. The work demonstrates that the PAH functionalized Pt NLAs are indeed promising cathodic electrocatalysts for practical application in direct alcohol fuel cells.


Intraarterial chemotherapy through carotid transposition in advanced head and neck cancer.
We report results of intraarterial chemotherapy (IAC) in patients with advanced unresectable (T4, N0-3, MO) squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) of the oral cavity, oropharynx, or facial skin, given through a subcutaneously transposed external carotid artery by a fine gauge needle. Forty patients entered the study and 39 were evaluable for clinical response and toxicity. The infusion technique was done in two consecutive series of patients with the same characteristics: 17 had treatment with vincristine, bleomycin, and methotrexate (VBM), and 22 received cisplatin and bleomycin (PB) with a median number of six and five cycles, respectively. The total objective response (OR) rate was 76.9%, which included 12.8% of complete response (CR). A better percentage of OR was attained in BCC than SCC (83.3% versus 75.8%), in tumors of the skin of the face than other sites (87.5% versus 74.2%), and in the group that received PB than the group that received VBM (77.3% versus 76.5%). Our series had an overall technique-related toxicity of 18%; 10.3% with neurologic disturbances and 7.7% with skin inflammation (almost all were reversible). The most common systemic toxic side effects were generally mild and well tolerated (30.8%), whereas major toxicities (severe modifications of the cardiopulmonary function) occurred in four patients (10.3%) in the group that received VBM. After chemotherapy, 28 patients (71.8%) were deemed resectable; 23 of these patients (82.1%) agreed to surgery. Pathologic CR were obtained in 13 patients (56.5%). Our technique has significant advantages over the conventional methods of infusion, because tumor perfusion is feasible and selective, technique-related and catheter-related complications are almost completely eliminated, and fatal drug-related side effects are avoided.


A population-based cohort study of pregnancy outcomes among women with primary sclerosing cholangitis.
Studies of primary sclerosing cholangitis (PSC) and pregnancy outcomes have been limited in size and have been inadequate to rule out excess risks. We examined pregnancy outcomes among women with PSC. Women with PSC were identified from inpatient and hospital-based outpatient data in the Swedish National Patient Register. Through linkage with the Medical Birth Register, we identified 229 singleton births, from 1987 through 2009, to women with PSC before delivery. These were compared with 2,304,863 births to women without a diagnosis of PSC. We used logistic regression, adjusted for maternal age, smoking, education, parity, and year of birth, to calculate adjusted prevalence odds ratios (aPORs) for adverse pregnancy outcomes. Maternal PSC was associated with a 3.63-fold increase in preterm birth (95% confidence interval [CI] for aPOR, 2.35-5.61) as well as an increased risk of cesarean section (aPOR, 2.18; 95% CI, 1.50-3.17). We found no increased risk based on analyses of the 5-minute Apgar score, small for gestational age, stillbirths, or neonatal deaths. Maternal PSC was not a risk factor for congenital abnormalities (aPOR, 1.12; 95% CI, 0.56-2.22). Stratification by inflammatory bowel disease status did not affect the risk estimates more than marginally. Maternal PSC is associated with both preterm birth and cesarean section but not with congenital malformation or other adverse outcomes of pregnancy. Pregnancy should not be discouraged in women with PSC.


Azacitidine for treatment of patients with myelodysplastic syndromes (MDS): practical recommendations of the German MDS Study Group.
Myelodysplastic syndromes (MDS) are a group of common bone marrow disorders characterized by ineffective hematopoiesis, peripheral cytopenias, and a substantial risk of progression to acute myeloid leukemia (AML). For many years, the main treatment option for MDS was best supportive care which alleviates symptoms, but has no effect on the natural course of the disease. Recently, demethylating agents have become available as a promising new treatment for patients with MDS. In two randomized clinical trials, the demethylating agent azacitidine has demonstrated a reduced risk of transformation to AML, improvement of peripheral blood values, an improved quality of life, and a definite survival advantage compared to conventional care regimens for patients with International Prognostic Scoring System score of intermediate-2 or high-risk MDS. This review aims to provide practical recommendations for the use of azacitidine and the management of its side effects in patients with MDS, assuring safe administration and best efficacy of treatment.


Pharmacokinetics, Safety and Efficacy of Maraviroc in Treatment-experienced Pediatric Patients Infected With CCR5-Tropic HIV-1.
Maraviroc is a CC-chemokine receptor 5 antagonist approved to treat adults infected with CC-chemokine receptor 5-tropic (R5) HIV-1. Study A4001031 was conducted to evaluate the pharmacokinetics, safety and efficacy of maraviroc in combination with optimized background therapy in treatment-experienced pediatric patients infected with R5 HIV-1 and support registration of maraviroc for pediatric use. This is an open-label, 2-stage, age-stratified, noncomparative multicenter study. One-hundred and three participants were enrolled into 4 age/formulation cohorts and dosed twice daily. Initial doses were determined by body surface area and optimized background therapy, based on drug interactions with maraviroc in adults. Dose adjustment and pharmacokinetic reevaluation occurred if the average concentrations (Cavg) at Week 2 were <100 ng/mL (Stage 1-dose finding). Data from the Week 48 analysis demonstrated that 49/50 Stage 1 participants rolling over into Stage 2 (safety and efficacy) achieved Cavg ≥100 ng/mL. Doses were identified that achieved similar concentration ranges to those seen in adults. The majority (90/103) received optimized background therapy containing potent cytochrome P450 3A inhibitors. Maraviroc was well tolerated and the safety and efficacy were comparable to those of adults. All cohorts had a mean decrease from baseline in HIV-1 RNA of >1 log10. Increases from baseline in the median CD4+ cell count and percentage were seen for all age groups. The maraviroc dosing strategy resulted in participants achieving the target Cavg, with exposure ranges similar to those observed in adults on approved doses. The safety and efficacy of maraviroc in this pediatric population were comparable to those seen in adults.


Too good for this world: moral bioenhancement and the ethics of making moral misfits.
Persson and Savulescu argue that moral bioenhancement is not only morally permissible; in some cases, it is morally obligatory. In this article, I introduce a new reason to worry about moral enhancement. I adapt the disability concept of misfit to show how moral enhancement could cause extreme moral disempowerment to those enhanced, which would result in moral injury. I argue that any safety framework that guides the development of moral bioenhancement must be sensitive to the problem of moral misfitting. I present the best case for moral bioenhancement before turning to my own worry concerning the development of moral bioenhancement and its practical implications. Finally, I consider a series of objections and responses.


Growth, puberty and reproduction in gilts born to dams deprived of nutrients during pregnancy.
Reproductive performance through two gestations was evaluated in a random selection of gilts born to dams deprived of nutrients or full-fed during pregnancy. After weaning, gilts born to dams previously subjected to inanition during the middle third or last third of gestation (O kcal/d; water only; 40 d), and those born to full-fed (7.028) kcal/d) control dams throughout gestation were raised in the same outdoor lot. At approximately 150 d of age, the gilts were observed for estrous behavior and mated at the third estrus. Age at puberal estrus was earlier (P less than .01) for gilts born to nutrient-deprived dams. Gilts from control dams exhibited first estrus at 250 d, whereas puberal estrus in gilts born to dams deprived of nutrients during the middle or last third of pregnancy occurred at 208 and 219 d, respectively. Fifteen of 18 mated gilts from the nutrient-deprived dams remained pregnant to term, compared with seven of eight gilts from control dams. Birth weights of first-parity living neonates from gilts born to nutrient-deprived dams were similar to those of newborns from controls. First-parity litter weights at birth also were similar for the experimental and control groups. Litter size at birth of first-parity living neonates averaged 7.9 piglets for gilts born to nutrient-deprived dams and was similar (P less than .05) to that for gilts from full-fed controls (9.4 piglets). Second-parity pregnancy rate, birth weight, litter weight and litter size at birth and weaning were similar for all gilts born to nutrient-deprived and control dams. These results indicate that brief segments of maternal nutrient deprivation during middle or late pregnancy have little detrimental effect on subsequent growth and reproduction of the female progeny in this litter-bearing species.


Distributions of immunoglobulin-containing cells in alimentary tract, spleen, and mesenteric lymph node of the pig demonstrated by peroxidase-conjugated antiserums to porcine immunoglobulins G, A, and M.
Peroxidase-conjugated antibody to porcine immunoglobulins A, G, and M were used in a light microscope study of the distribution of immunoglobulins and immunoglobulin-containing cells in alimentary tract, spleen, and mesenteric lymph node of the pig. Largest populations of plasma cells staining for each of the 3 immunoglobulin classes were demonstrated in the lamina propria of the intestinal tract. Immunoglobulin A cells were most numerous, and immunoglobulin A and M cells each outnumbered immunoglobulin G cells which were, however, present in considerable numbers. Both immunoglobulins A and M were demonstrated in intestinal crypt epithelial cells, indicating they have similar intracellular secretion pathways. Porcine spleen and mesenteric lymph node seem to have a minor role in immunoglobulin synthesis.


Renal histopathology in hypertensive diabetic patients.
A total of 250 renal biopsy specimens from diabetic patients and from the kidneys of 400 autopsy cases were examined histologically and compared to kidneys from 160 autopsied nondiabetics. The morphological findings were assessed in relation to hypertension. There was a high prevalence of arteriolosclerosis, glomerulosclerosis, and pyelonephritis; in addition, early diabetic glomerulopathy and glomerulonephritis, particularly of the membranous type, were noted in a remarkably high percentage of diabetic patients. Ninety-three percent of patients with hypertension had arteriolosclerosis, and a good correlation existed between the extent of this lesion and the level of blood pressure. Even in 66% of normotensive patients, however, arteriolosclerosis was found. This fact and the involvement of the vas efferens argue against the notion of arteriolosclerosis being exclusively a sequela of hypertension. More than 70% of patients with glomerulosclerosis suffered from hypertension, compared to less than 50% of patients without either that condition or early diabetic lesions. The majority of diabetic patients with pyelonephritis and glomerulonephritis were hypertensive. We conclude that hypertension in diabetic patients with renal involvement may result from different renal lesions that can be differentiated only by histological examination.


Results of mental support for health care professionals and mental care during the COVID-19 pandemic.
The coronavirus disease 2019 (COVID-19) resulted into a global pandemic and continues to thrive until vaccines have been successfully developed and distributed around the world. The outcomes of COVID-19 contaminations range from death to minor health-related complaints. Furthermore, and not less significant, the increasing pressure on local as well as global health care is rising. In The Netherlands but also in other countries, further intensified regulations are introduced in order to contain the second wave of COVID-19, primarily to limit the number contaminations but also to prevent the health care professionals for giving in to the rising pressure on them. The results of the campaign for health care professionals in The Netherlands show that health care professionals are increasingly searching for information regarding psychological symptoms such as feeling of uncertainty, pondering and advice regarding the support of care teams. In this short update, we provide the results of the previous campaign and stress the importance of support after COVID-19 based on these results.


Abortion and early neonatal death of kids attributed to intrauterine Yersinia pseudotuberculosis infection.
Yersinia pseudotuberculosis was isolated from placenta and abomasal contents of triplet goat kids, two of which were aborted and one of which died shortly after birth. Necropsy findings in the kids were suppurative placentitis and suppurative pneumonia. The public health implications in intrauterine Yersinia infection in goats are discussed.


Risk-stratified imputation in survival analysis.
Censoring that is dependent on covariates associated with survival can arise in randomized trials due to changes in recruitment and eligibility criteria to minimize withdrawals, potentially leading to biased treatment effect estimates. Imputation approaches have been proposed to address censoring in survival analysis; while these approaches may provide unbiased estimates of treatment effects, imputation of a large number of outcomes may over- or underestimate the associated variance based on the imputation pool selected. We propose an improved method, risk-stratified imputation, as an alternative to address withdrawal related to the risk of events in the context of time-to-event analyses. Our algorithm performs imputation from a pool of replacement subjects with similar values of both treatment and covariate(s) of interest, that is, from a risk-stratified sample. This stratification prior to imputation addresses the requirement of time-to-event analysis that censored observations are representative of all other observations in the risk group with similar exposure variables. We compared our risk-stratified imputation to case deletion and bootstrap imputation in a simulated dataset in which the covariate of interest (study withdrawal) was related to treatment. A motivating example from a recent clinical trial is also presented to demonstrate the utility of our method. In our simulations, risk-stratified imputation gives estimates of treatment effect comparable to bootstrap and auxiliary variable imputation while avoiding inaccuracies of the latter two in estimating the associated variance. Similar results were obtained in analysis of clinical trial data. Risk-stratified imputation has little advantage over other imputation methods when covariates of interest are not related to treatment. Risk-stratified imputation is intended for categorical covariates and may be sensitive to the width of the matching window if continuous covariates are used. The use of the risk-stratified imputation should facilitate the analysis of many clinical trials, in which one group has a higher withdrawal rate that is related to treatment.


The therapeutic potential of inhibitors of dipeptidyl peptidase IV (DPP IV) and related proline-specific dipeptidyl aminopeptidases.
In this review the structural and functional aspects of dipeptidyl peptidase IV (DPP IV) will be described, and the therapeutic potential of DPP IV inhibitors will be highlighted. DPP IV will be situated in clan SC, a group of serine proteases that contains several proline specific peptidases. Structural aspects of DPP IV and its interaction with different types of inhibitors are recently revealed by the publication of several crystal structures. Especially the design and development of new DPP IV inhibitors based on the three-dimensional structure, substrate specificity and catalytic mechanism will be discussed. In the last years there was an important development of new pyrrolidine-2-nitriles with very promising therapeutic properties for the treatment of type 2 diabetes. The role of DPP IV in peptide metabolism of members of the PACAP/glucagon peptide family, neuropeptides and chemokines has been thoroughly investigated during recent years. This is directly related to the promising therapeutic potential of DPP IV inhibitors in the treatment of type 2 diabetes and in the treatment of immunological disorders. Several inhibitors are currently under investigation in clinical trials for the treatment of type 2 diabetes and represent a new class of drugs for the treatment of this disease.


Clinically indicated electrical stimulation strategies to treat patients with medically refractory epilepsy.
Focal epilepsies represent approximately half of all diagnoses, and more than one-third of these patients are refractory to pharmacologic treatment. Although resection can result in seizure freedom, many patients do not meet surgical criteria, as seizures may be multifocal in origin or have a focus in an eloquent region of the brain. For these individuals, several U.S. Food and Drug Administration (FDA)-approved electrical stimulation paradigms serve as alternative options, including vagus nerve stimulation, responsive neurostimulation, and stimulation of the anterior nucleus of the thalamus. All of these are safe, flexible, and lead to progressive seizure control over time when used as an adjunctive therapy to antiepileptic drugs. Focal epilepsies frequently involve significant comorbidities such as cognitive decline. Similar to antiepilepsy medications and surgical resection, current stimulation targets and parameters have yet to address cognitive impairments directly, with patients reporting persistent comorbidities associated with focal epilepsy despite a significant reduction in the number of their seizures. Although low-frequency theta oscillations of the septohippocampal network are critical for modulating cellular activity and, in turn, cognitive processing, the coordination of neural excitability is also imperative for preventing seizures. In this review, we summarize current FDA-approved electrical stimulation paradigms and propose that theta oscillations of the medial septal nucleus represent a novel neuromodulation target for concurrent seizure reduction and cognitive improvement in epilepsy. Ultimately, further advancements in clinical neurostimulation strategies will allow for the efficient treatment of both seizures and comorbidities, thereby improving overall quality of life for patients with epilepsy.


Prophage induction in lysogenic Escherichia coli with simple hydroxylamine and hydrazine compounds.
The prophage-inducing capability of hydroxylamine sulfate and 36 of its derivatives, and of hydrazine dihydrochloride and dihydrazine sulfate and 43 of their derivatives, was determined in Escherichia coli W1709 (lambda). Maximal nontoxic concentrations up to 1 mg/ml were tested. Hydroxylamine sulfate was active at 2.5 mug/ml and the following 17 derivatives were active at concentrations ranging up to 500 mug/ml: alpha-naphthylhydroxylamine, N-hydroxy-2-aminofluorene, oxamyl hydroxamic acid, O-carbamoyl hydroxylamine (isohydroxyurea), N-hydroxyurethane, N-methylhydroxylamine HCl, salicylhydroxamic acid, oxalohydroxamic acid, methoxyamine HCl, ethoxyamine HCl, N, N-diethylhydroxylamine oxalate, formaldoxime, formamidoxime, acetohydroxamic acid, acetaldoxime, acetone oxime, and hydroxyguanidine sulfate. Hydrazine dihydrochloride and dihydrazine sulfate were effective inducers at 5.0 and 2.5 mug/ml, respectively, and the following nine derivatives of them were active at concentrations ranging up to 500 mug/ml: phthalic acid hydrazide, phenylhydrazine HCl, p-nitrophenylhydrazine, p-chlorophenylhydrazine HCl, formylhydrazine, carbohydrazide, semicarbazide HCl, 1-methyl-1-phenyl-hydrazine sulfate, and acetic acid hydrazide. Nineteen hydroxylamine and 34 hydrazine derivatives were ineffective as inducers. Application of the prophage-induction system as a tool for detection of responsive hydroxylamino and hydrazino compounds which may be potential toxicological hazards in the environment is discussed.


New viruses described in finfish from 1988-1992.
A number of new virus isolates from finfish have been reported in the scientific literature during the past five years. These include nine aquareoviruses, eight picornaviruses, six iridoviruses, five herpesviruses, three rhabdoviruses, three retroviruses, a paramyxovirus, and a coronavirus. Not all of these agents have been isolated in cell culture or established as etiologic agents of disease by controlled transmission studies. The burgeoning number of fish viruses is a reflection of the increased interest in fish diseases, particularly those occurring in aquaculture facilities, and the number will surely grow as fish farming intensifies on a global scale. This review chronicles the new virus isolates and lists them with other members of their virus family where appropriate.


A comparison of flight and non-flight sick call visits to a U.S. Army aviation medicine clinic.
In a U.S. Army aviation medicine program, flight surgeons provide primary clinical care for the units they support. Routine outpatient care (sick call) is performed daily on a walk-in basis. This study describes the diagnoses associated with flight and non-flight personnel reporting to sick call in an Army aviation medicine clinic. Data were collected over a 1-year period at an Aviation Troop Medical Clinic (TMC) supporting an aviation brigade consisting of 530 aircrew and 1,220 support personnel. A total of 5,793 sick call visits were made, 73% of which were initial visits, the remainder being second or subsequent visits. Aircrew evaluations accounted for 29% of the total. Major reasons for visits were illness (56%), injury (34%), and supplemental care (10%). However, substantial differences were noted between flight and non-flight patients. Overall, non-aircrew visits accounted for the bulk of the flight surgeons' clinical workload during sick call.


Risk reduction and health promotion behaviors following genetic testing for adult-onset disorders.
Although clinical genetic testing is available for over 1,000 inherited disorders, consequences of predictive genetic testing have been most extensively examined for hereditary breast and ovarian cancer (HBOC), hereditary colon cancer, and Huntington disease (HD). These focus primarily on psychological, ethical, legal, and social aspects of genetic testing. Genetic testing may also provide information that can lead to behaviors that promote health and reduce risk for disease, reflecting options available for the disorder for which the person is at risk. However, regardless of condition, people completing a genetic test may inform relatives about the results of the test and implications for their risk to develop the condition. Literature on risk reduction behaviors and communication focuses on families with HBOC or colorectal cancer. Few reports document behaviors for other conditions. This paper presents a systematic review of the research literature on risk reduction and health promotion behaviors following clinical genetic testing for adult onset conditions, primarily HBOC, familial colon cancers, and HD. Insights gleaned from this review are discussed as a basis for planning monitoring of health promotion and risk-reduction behaviors for genetic testing for present and future use.


Absorption and pharmacokinetics of phenylbutazone in Welsh Mountain ponies.
The disposition of phenylbutazone (4.4 mg/kg), administered intravenously to six Welsh Mountain ponies, was described by a two-compartment open model. Pharmacokinetic parameters were not significantly different after morning dosing in comparison with afternoon dosing. When phenylbutazone (4.4 mg/kg) was administered orally to the same ponies, marked variations in time to peak concentrations were produced with different feeding schedules. When access to hay was permitted before and after dosing, the mean time to peak concentration was 13.2 +/- 1.2 h and double peaks in the plasma concentration-time curve were common. Double peaks were also encountered when phenylbutazone was given to ponies deprived of food prior to, and allowed access to hay after, dosing. In this circumstance, mean times to peak concentration were much shorter (3.8 +/- 1.3 h after morning dosing and 5.3 +/- 1.5 h followed afternoon dosing). Absorption was more regular and double peaks were less apparent when food was withheld both before and after dosing. In order to explain these findings, it is tentatively postulated that, whereas some of the administered dose of phenylbutazone may be absorbed quickly, some may become adsorbed on to the feed and subsequently released by fermentative digestion in the large intestine and/or caecum. The consequences of delayed absorption in fed animals for toxicity and clinical efficacy, and for the use of phenylbutazone in equestrian sports, are considered. Delayed absorption in ponies given access to hay was not accompanied by a significant reduction in total absorption. Bioavailability was estimated to be approximately 69% in fed and 78% in unfed ponies. Estimates of bioavailability gave similar values for morning (72%) and afternoon (71%) dosing.


A new human cytomegalovirus isolate has an invertible subsegment within its L component producing eight genome isomers.
A HindIII cleavage map of the genome DNA of a new isolate of human cytomegalovirus (HCMV), strain Tanaka, was constructed by cosmid cloning and Southern blot hybridization of virion DNA. The genome was found to be unique in that its long (L) component was composed of two subsegments, L1 and L2, and subsegment L2 underwent inversion relative to L1 at high frequency. In addition to the normal inversions of the L and short (S) components, this produced eight genome isomers. The novel invertible subsegment was flanked by an inverted sequence distinct from the inversion-specific a sequence present in the terminal and junction regions of the genome.


Human Neural Stem Cell Systems to Explore Pathogen-Related Neurodevelopmental and Neurodegenerative Disorders.
Building and functioning of the human brain requires the precise orchestration and execution of myriad molecular and cellular processes, across a multitude of cell types and over an extended period of time. Dysregulation of these processes affects structure and function of the brain and can lead to neurodevelopmental, neurological, or psychiatric disorders. Multiple environmental stimuli affect neural stem cells (NSCs) at several levels, thus impairing the normal human neurodevelopmental program. In this review article, we will delineate the main mechanisms of infection adopted by several neurotropic pathogens, and the selective NSC vulnerability. In particular, TORCH agents, i.e., Toxoplasma gondii, others (including Zika virus and Coxsackie virus), Rubella virus, Cytomegalovirus, and Herpes simplex virus, will be considered for their devastating effects on NSC self-renewal with the consequent neural progenitor depletion, the cellular substrate of microcephaly. Moreover, new evidence suggests that some of these agents may also affect the NSC progeny, producing long-term effects in the neuronal lineage. This is evident in the paradigmatic example of the neurodegeneration occurring in Alzheimer's disease.


Regional variation in otolith Sr:Ca ratios of African longfinned eel Anguilla mossambica and mottled eel Anguilla marmorata: a challenge to the classic tool for reconstructing migratory histories of fishes.
Otolith Sr:Ca ratios of the African longfinned eel Anguilla mossambica and giant mottled eel Anguilla marmorata from nine freshwater sites in four rivers of South Africa were analysed to reconstruct their migratory life histories between freshwater and saltwater habitats. For A. mossambica, the Sr:Ca ratios in the otolith edge differed significantly among rivers and had large effect sizes, but did not differ among sites within a river. Otolith Sr:Ca ratios did not differ among rivers for A. marmorata. When rivers were pooled, the edge Sr:Ca ratios of A. mossambica were not significantly different from those of A. marmorata. According to the river-specific critical Sr:Ca ratio distinguishing freshwater from saltwater residence, most A. mossambica and A. marmorata had saltwater habitat experience after settlement in fresh water. This was primarily during their elver stage or early in the yellow eel stage. During the middle and late yellow eel stage, freshwater residency was preferred and only sporadic visits were made to saltwater habitats. The data also suggest that regional variations in otolith Sr:Ca ratios affect the critical Sr:Ca value and are a challenge for the reconstruction of migratory life histories that should be explicitly considered to avoid bias and uncertainty.


A Novel Liquid-Liquid Extraction for the Determination of Nicotine in Tap Water, Wastewater, and Saliva at Trace Levels by GC-MS.
Determination of nicotine at trace levels in different matrixes is crucial because of nicotine's strongly addictive property. In this study, a simple and fast analytical method was developed using liquid- liquid extraction with GC-MS. Chloroform was used as an extraction solvent. Different parameters for extraction, such as pH of the solution, types and volumes of extraction and supporter solvents for dispersion, extraction period, and salt type and its amount, were optimized. To improve the precision, naphthalene was used as an internal standard. The calibration plot of nicotine was linear from 0.010 to 2.0 mg/L with a correlation coefficient 0.9996. The LOD and LOQ for nicotine after extraction were 2.6 and 8.8 ng/mL, respectively. Under the optimum conditions, tap water, wastewater, and saliva samples were analyzed for their nicotine content. The spiking experiments yielded satisfactory recoveries of 97.6 ± 3.5, 96.8 ± 1.1, and 85.1 ± 1.3% for tap water, wastewater, and saliva samples, respectively.


Role of oxidized low density lipoprotein in atherogenesis.
Evidence to support an important role of oxidative modification in mediating the atherogenicity of LDL continues to grow. New hypotheses suggest mechanisms by which Ox-LDL or products of Ox-LDL can affect many components of the atherogenic process, including vasomotor properties and thrombosis, as well as lesion initiation and progression itself. These ideas suggest new approaches, that in combination with lowering of plasma cholesterol, could lead to the prevention of atherosclerosis and its complications.


Recurrent alien hand syndrome in a multiple sclerosis case.
Alien hand syndrome is the strange feeling of one's hand behaving independently. This syndrome has rarely been reported in multiple sclerosis (MS) patients. Herein, we present a 34-year-old female MS patient who had recurrent symptoms of alien hand syndrome that were evaluated as MS attacks based on cranial magnetic resonance imaging that showed demyelinating lesions in the corpus callosum. Alien hand syndrome is classified according to the location of the lesion and the presenting symptoms. As such, our patient can best be classified as a callosal alien hand case.


[Effect of neuropeptide fragments of adrenocorticotropic hormone and vasopressin on the behavioral activity of rats].
The effects of vasopressin and those of fragments of adrenocorticotropin ACTH5-10; ACTH4-7; l-phenilalanin, and ACTH4-7 d-phenilalanin on the learning of rats in U-shaped labyrinth were studied. The consolidation of skills in long-term memory ability for reversal learning were checked. The results suggest that effect of all these oligopeptides on the higher nervous activity is significant and specific, vasopressin improving the long-term memory, ACTH4-7-l-phenilalanin improving the learning, ACTH4-7-d-phenilalanin making learning next to impossible.


Evidence-Based Treatment Options in Recurrent and/or Metastatic Squamous Cell Carcinoma of the Head and Neck.
The major development of the past decade in the first-line treatment of recurrent and/or metastatic squamous cell carcinoma of the head and neck (R/M SCCHN) was the introduction of cetuximab in combination with platinum plus 5-fluorouracil chemotherapy (CT), followed by maintenance cetuximab (the "EXTREME" regimen). This regimen is supported by a phase 3 randomized trial and subsequent observational studies, and it confers well-documented survival benefits, with median survival ranging between approximately 10 and 14 months, overall response rates between 36 and 44%, and disease control rates of over 80%. Furthermore, as indicated by patient-reported outcome measures, the addition of cetuximab to platinum-based CT leads to a significant reduction in pain and problems with social eating and speech. Conversely, until very recently, there has been a lack of evidence-based second-line treatment options, and the therapies that have been available have shown low response rates and poor survival outcomes. Presently, a promising new treatment option in R/M SCCHN has emerged: immune checkpoint inhibitors (ICIs), which have demonstrated favorable results in second-line clinical trials. Nivolumab and pembrolizumab are the first two ICIs that were approved by the US Food and Drug Administration. We note that the trials that showed benefit with ICIs included not only patients who previously received ≥1 platinum-based regimens for R/M SCCHN but also patients who experienced recurrence within 6 months after combined modality therapy with a platinum agent for locally advanced disease. In this review, we outline the available clinical and observational evidence for the EXTREME regimen and the initial results from clinical trials for ICIs in patients with R/M SCCHN. We propose that these treatment options can be integrated into a new continuum of care paradigm, with first-line EXTREME regimen followed by second-line ICIs. A number of ongoing clinical trials are comparing regimens with ICIs, alone and in combination with other ICIs or CT, with the EXTREME regimen for first-line treatment of R/M SCCHN. As we eagerly await the results of these trials, the EXTREME regimen remains the standard of care for the first-line treatment of R/M SCCHN.


Chemical Mimicry in the Myrmecophilous Beetle Myrmecaphodius excavaticollis.
The myrmecophilus beetle Myrmecaphodius excavaticollis (Blanchard) was found to have species-specific cuticular hydrocarbons acquired-from one of its hosts, the ant Solenopsis richteri Forel. Removal from its ant host resulted in loss of the host hydrocarbons, leaving a cuticular pattern innate to the beetle. When beetles were transferred to colonies of three other Solenopsis species, they acquired the specific hydrocarbons associated with each of the new hosts. This passive integration mechanism is coupled with the beetle's armored exterior to enable it to cope with multiple aggressive hosts.


Anatabine, Nornicotine, and Anabasine Reduce Weight Gain and Body Fat through Decreases in Food Intake and Increases in Physical Activity.
Obesity is a leading cause of preventable death in the United States. Currently approved pharmacotherapies for the treatment of obesity are associated with rebound weight gain, negative side effects, and the potential for abuse. There is a need for new treatments with fewer side effects. Minor tobacco alkaloids (MTAs) are potential candidates for novel obesity pharmacotherapies. These alkaloids are structurally related to nicotine, which can help reduce body weight, but without the same addictive potential. The purpose of the current study was to examine the effects of three MTAs (nornicotine, anatabine, and anabasine) and nicotine on weight gain, body composition, chow intake, and physical activity. We hypothesized that the MTAs and nicotine would reduce weight gain through reductions in chow intake and increases in physical activity. To test this, male Sprague Dawley rats were housed in metabolic phenotyping chambers. Following acclimation to these chambers and to (subcutaneous (sc)) injections of saline, animals received daily injections (sc) of nornicotine, anabasine, anatabine, or nicotine for one week. Compared to saline-injected animals that gained body weight and body fat during the treatment phase, injections of nornicotine and anatabine prevented additional weight gain, alongside reductions in body fat. Rats receiving anabasine and nicotine gained body weight at a slower rate relative to rats receiving saline injections, and body fat remained unchanged. All compounds reduced the intake of chow pellets. Nornicotine and nicotine produced consistent increases in physical activity 6 h post-injection, whereas anabasine's and anatabine's effects on physical activity were more transient. These results show that short-term, daily administration of nornicotine, anabasine, and anatabine has positive effects on weight loss, through reductions in body fat and food intake and increases in physical activity. Together, these findings suggest that MTAs are worthy of further investigations as anti-obesity pharmacotherapies.


Cellular binding proteins for vitamin A in human carcinomas and in normal tissues.
Blinded analyses of the concentrations of binding proteins for retinol and retinoic acid (CRABP) in homogenates of cancer and normal tissue aliquots obtained from human cervix, endometrium, ovary, breast, and lung were carried out by the sucrose gradient ultracentrifugation technique. In carcinomas of the cervix and endometrium, CRABP mean values of 50.4 and 123.2 pmol/g tissue, respectively were detected. Such concentrations represent a 3- and 4-fold increase over the mean values of CRABP in the normal cervix (16.9 pmol/g) and normal endometrium (30.8 pmol/g), respectively. In carcinomas of the ovary, the mean CRABP level was 128.6 pmol/g compared to the maximal mean value of less than or equal to 0.46 pmol/g in the normal ovary. Elevated levels of CRABP were also found in breast and lung carcinomas compared to the amounts detected in the same patient in normal tissue aliquots of the same organ. The differences between CRABP concentrations in cervical, endometrial, ovarian, and breast carcinomas and those in normal tissue are statistically significant. In contrast, cellular retinol-binding protein concentrations were reduced in the endometrial, ovarian, breast, and lung carcinomas compared to normal tissues. There were no significant differences between the log-mean concentrations of cellular retinol-binding proteins in the cytosols from tissue aliquots of carcinoma of the cervix and those in the cytosols from tissue aliquots of normal cervix.


Development and evaluation of a laparoscopic common bile duct exploration simulator and procedural rating scale.
Laparoscopic common bile duct exploration (LCBDE) is an effective, single-stage treatment for choledocholithiasis. However, LCBDE requires specific cognitive and technical skills, is infrequently performed by residents, and currently lacks suitable training and assessment modalities outside of the operating room. To address this gap in training, a simulator model for transcystic and transcholedochal LCBDE was developed and evaluated. A procedure algorithm incorporating essential cognitive and technical steps of LCBDE was developed, along with a physical model to allow performance of a simulated procedure. Modified Objective Structured Assessment of Technical Skills (OSATS) rating scales were developed to assess performance on the model. Construct validity was assessed by comparing the performance of novices (residents and surgeons without LCBDE experience) versus experienced subjects (surgeons with previous LCBDE experience). Concurrent validity was assessed by comparing scores from the LCBDE scales to those from the standard OSATS scale. Internal consistency and interrater reliability were assessed by comparing performance scores assigned by three independent raters. Sixteen novices and five experienced subjects performed simulated procedures, with novices scoring lower than experienced subjects on both transcystic (20 ± 3 vs. 33 ± 2 [possible score range, 0-45], p < 0.001) and transcholedochal (25 ± 8 vs. 42 ± 3 [possible score range, 0-53], p < 0.001) rating scales. Scores on the rating scales correlated significantly with scores from the standard OSATS scale. Internal consistency and interrater reliability of the LCBDE rating scales were favorable. The LCBDE simulator is a low-cost yet realistic physical model that allows performance and evaluation of technical skills required for LCBDE. The LCBDE rating scales show evidence of construct validity, concurrent validity, internal consistency, and interrater reliability. Use of the LCBDE model and associated rating scales allows procedure-specific feedback for trainees and could be used to improve current training.


Influence of Local Muscle Cooling on Mitochondrial-Related Gene Expression at Rest.
The purpose of this study was to determine the impact of localized cooling of the skeletal muscle during rest on mitochondrial related gene expression. Thermal wraps were applied to the vastus lateralis of each limb of 12 participants. One limb received a cold application (randomized) (COLD), while the other did not (RT). Wraps were removed at the 4 h time point and measurements of skin temperature, blood flow, and intramuscular temperature were taken prior to a muscle biopsy. RT-qPCR was used to measure expression of genes associated with mitochondrial development. Skin and muscle temperatures were lower in COLD than RT (p < 0.05). Femoral artery diameter was lower in COLD after 4 h (0.62 ± 0.05 cm, to 0.60 ± 0.05 cm, p = 0.018). Blood flow was not different in COLD compared to RT (259 ± 69 mL·min-1 vs. 275 ± 54 mL·min-1, p = 0.20). PGC-1α B and GABPA expression was higher in COLD relative to RT (1.57-fold, p = 0.037 and 1.34-fold, p = 0.006, respectively). There was no difference (p > 0.05) in the expression of PGC-1α, NT-PGC-1α, PGC-1α A, TFAM, ESRRα, NRF1, GABPA, VEGF, PINK1, PARK 2, or BNIP3-L. The impact of this small magnitude of difference in gene expression of PGC-1α B and GABPA without alterations in other genes are unknown. There appears to be only limited impact of local muscle cooling on the transcriptional response related to mitochondrial development.


Tre1 GPCR initiates germ cell transepithelial migration by regulating Drosophila melanogaster E-cadherin.
Despite significant progress in identifying the guidance pathways that control cell migration, how a cell starts to move within an intact organism, acquires motility, and loses contact with its neighbors is poorly understood. We show that activation of the G protein-coupled receptor (GPCR) trapped in endoderm 1 (Tre1) directs the redistribution of the G protein Gbeta as well as adherens junction proteins and Rho guanosine triphosphatase from the cell periphery to the lagging tail of germ cells at the onset of Drosophila melanogaster germ cell migration. Subsequently, Tre1 activity triggers germ cell dispersal and orients them toward the midgut for directed transepithelial migration. A transition toward invasive migration is also a prerequisite for metastasis formation, which often correlates with down-regulation of adhesion proteins. We show that uniform down-regulation of E-cadherin causes germ cell dispersal but is not sufficient for transepithelial migration in the absence of Tre1. Our findings therefore suggest a new mechanism for GPCR function that links cell polarity, modulation of cell adhesion, and invasion.


Hierarchical folding mechanism of apomyoglobin revealed by ultra-fast H/D exchange coupled with 2D NMR.
The earliest steps in the folding of proteins are complete on an extremely rapid time scale that is difficult to access experimentally. We have used rapid-mixing quench-flow methods to extend the time resolution of folding studies on apomyoglobin and elucidate the structural and dynamic features of members of the ensemble of intermediate states that are populated on a submillisecond time scale during this process. The picture that emerges is of a continuum of rapidly interconverting states. Even after only 0.4 ms of refolding time a compact state is formed that contains major parts of the A, G, and H helices, which are sufficiently well folded to protect amides from exchange. The B, C, and E helix regions fold more slowly and fluctuate rapidly between open and closed states as they search docking sites on this core; the secondary structure in these regions becomes stabilized as the refolding time is increased from 0.4 to 6 ms. No further stabilization occurs in the A, G, H core at 6 ms of folding time. These studies begin to time-resolve a progression of compact states between the fully unfolded and native folded states and confirm the presence an ensemble of intermediates that interconvert in a hierarchical sequence as the protein searches conformational space on its folding trajectory.


Dopamine-galanin receptor heteromers modulate cholinergic neurotransmission in the rat ventral hippocampus.
Previous studies have shown that dopamine and galanin modulate cholinergic transmission in the hippocampus, but little is known about the mechanisms involved and their possible interactions. By using resonance energy transfer techniques in transfected mammalian cells, we demonstrated the existence of heteromers between the dopamine D(1)-like receptors (D(1) and D(5)) and galanin Gal(1), but not Gal(2) receptors. Within the D(1)-Gal(1) and D(5)-Gal(1) receptor heteromers, dopamine receptor activation potentiated and dopamine receptor blockade counteracted MAPK activation induced by stimulation of Gal(1) receptors, whereas Gal(1) receptor activation or blockade did not modify D(1)-like receptor-mediated MAPK activation. Ability of a D(1)-like receptor antagonist to block galanin-induced MAPK activation (cross-antagonism) was used as a "biochemical fingerprint" of D(1)-like-Gal(1) receptor heteromers, allowing their identification in the rat ventral hippocampus. The functional role of D(1)-like-Gal receptor heteromers was demonstrated in synaptosomes from rat ventral hippocampus, where galanin facilitated acetylcholine release, but only with costimulation of D(1)-like receptors. Electrophysiological experiments in rat ventral hippocampal slices showed that these receptor interactions modulate hippocampal synaptic transmission. Thus, a D(1)-like receptor agonist that was ineffective when administered alone turned an inhibitory effect of galanin into an excitatory effect, an interaction that required cholinergic neurotransmission. Altogether, our results strongly suggest that D(1)-like-Gal(1) receptor heteromers act as processors that integrate signals of two different neurotransmitters, dopamine and galanin, to modulate hippocampal cholinergic neurotransmission.


The direction and rate of bacteriophage T7 DNA packaging in vitro.
To determine the direction of the entry of DNA during in vitro bacteriophage T7 DNA packaging, incompletely packaged DNA (ipDNA) was fractionated by agarose gel electrophoresis after degradation of DNA outside of capsids and then release of packaged DNA from capsids. After fractionation, quantitative in-gel probing with a right end-specific oligonucleotide detects heterogeneous ipDNA (called right-end ipDNA). Most of the right-end ipDNA appears with kinetics expected of a precursor to the mature T7 DNA. In-gel probing with a left-end-specific oligonucleotide detects ipDNA (left-end ipDNA); the molar amount of left end ipDNA is always at least 50x less than the molar amount of right-end ipDNA. Left-end ipDNA appears with the kinetics of an abortive end product of T7 DNA packaging. Thus, productive T7 DNA packaging occurs in a right-to-left direction. Quantitation of the conversion of right-end ipDNA to mature-length DNA yields an estimate of the mean rate of right-to-left in vitro T7 DNA packaging: 28 +/- 6 kbp/min for the last 20-50% of the DNA packaged.


Significance of CD90+ cancer stem cells in human liver cancer.
This study characterized cancer stem cells (CSCs) in hepatocellular carcinoma (HCC) cell lines, tumor specimens, and blood samples. The CD90+ cells, but not the CD90(-) cells, from HCC cell lines displayed tumorigenic capacity. All the tumor specimens and 91.6% of blood samples from liver cancer patients bore the CD45(-)CD90+ population, which could generate tumor nodules in immunodeficient mice. The CD90+CD44+ cells demonstrated a more aggressive phenotype than the CD90+CD44(-) counterpart and formed metastatic lesions in the lung of immunodeficient mice. CD44 blockade prevented the formation of local and metastatic tumor nodules by the CD90+ cells. Differential gene expression profiles were identified in the CD45(-)CD90+ and CD45(-)CD90(-) cells isolated from tissue and blood samples from liver cancer patients and controls.


Limited efficacy of α-conopeptides, Vc1.1 and RgIA, to inhibit sensory neuron CaV current.
Chronic pain is very difficult to treat. Thus, novel analgesics are a critical area of research. Strong pre-clinical evidence supports the analgesic effects of α-conopeptides, Vc1.1 and RgIA, which block α9α10 nicotinic acetylcholine receptors (nAChRs). However, the analgesic mechanism is controversial. Some evidence supports the block of α9α10 nAChRs as an analgesic mechanism, while other evidence supports the inhibition of N-type CaV (CaV2.2) current via activation of GABAB receptors. Here we reassess the effect of Vc1.1 and RgIA on CaV current in rat sensory neurons. Unlike the previous findings, we found highly variable effects among individual sensory neurons, but on average only minimal inhibition induced by Vc1.1, and no significant effect on the current by RgIA. We also investigated the potential involvement of GABAB receptors in the Vc1.1 induced inhibition, and found no correlation between the size of CaV current inhibition induced by baclofen (GABAB agonist) vs. that induced by Vc1.1. Thus, GABAB receptors are unlikely to mediate the Vc1.1 induced CaV current inhibition. Based on the present findings, CaV current inhibition in dorsal root ganglia is unlikely to be the predominant mechanism by which either Vc1.1 or RgIA induce analgesia. Better analgesic drugs are desperately needed to help physicians to treat pain. While many pre-clinical studies support the analgesic effects of α-conopeptides, Vc1.1 and RgIA, the mechanism is controversial. The development of improved α-conopeptide analgesics would be greatly facilitated by a complete understanding of the analgesic mechanism. However, we show that we cannot reproduce one of the proposed analgesic mechanisms, which is an irreversible inhibition of CaV current in a majority of sensory neurons.


Effects of copper and zinc sources and inclusion levels of copper on weanling pig performance and intestinal microbiota.
A 42-d experiment was conducted to evaluate the effect of Cu and Zn source and Cu level on pig performance, mineral status, bacterial modulation, and the presence of antimicrobial-resistant genes in isolates of Enterococcus spp. At weaning, 528 pigs (5.9 ± 0.50 kg) were allotted to 48 pens of a randomized complete block design in a 2 × 2 factorial arrangement with two Cu and Zn sources (SF: sulfate and HCl: hydroxychloride) and two Cu levels (15 and 160 mg/kg). As a challenge, the pigs were reared in dirty pens used by a previous commercial batch. Two-phase diets were offered: the pre-starter (PS) phase from day 1 to 14 and the starter phase (ST) from day 14 to 42. At days 14 and 42, pigs were individually weighed and blood samples from one pig per pen were taken. At the end of the experiment, one pig per pen was euthanized to collect the samples. Feeding high levels of Cu increased body weight (BW) from 16.6 to 17.7 kg (P < 0.001). Furthermore, average daily gain, gain to feed (G:F) ratio, average daily feed intake (ADFI), and mineral status were enhanced with Cu at 160 mg/kg (P < 0.05) compared with Cu at 15 mg/kg. There was no effect of the interaction between source × level on any of the growth performance responses except for ADFI (P = 0.004) and G:F (P = 0.029) at the end of the ST period and for G:F (P = 0.006) for entire nursery period (day 0 to 42). At the end of the ST period, pigs fed Cu at 160 mg/kg as HCl had not only higher ADFI but also lower G:F than those fed Cu as SF at 160 mg/kg. Meanwhile, for the entire nursery period, G:F did not differ between pigs fed Cu at 160 mg/kg as HCl or SF. In colonic digesta, the relative abundance of Streptococcus, Enterobacter, Escherichia, among others, decreased (P-adjust < 0.05), while Lachnospira and Roseburia tended (P-adjust < 0.10) to increase in pigs fed Cu at 160 mg/kg as HCl compared with those fed Cu SF at 160 mg/kg. An increase (P-adjust < 0.05) in Methanosphaera and Roseburia was observed in pigs fed Cu at 160 mg/kg. From colon digesta, Enterococcus spp. was isolated in 40 samples, being E. faecalis the most dominating (65%) regardless of the experimental diet. Genes of ermB (7.5%) and tetM (5%) were identified. No genes for Cu (tcrB) or vancomycin (vanA, vanB, vanC1, and vanC2) were detected. In conclusion, European Union permissible levels of Cu (160 mg/kg), of both sources, were able to increase performance, mineral status, and bacterial modulation compared with nutritional level. Different effects on growth performance, mineral tissue content, and microbial modulation were observed between Cu and Zn sources.


Fucosylated inhibitors of recently identified bangle lectin from Photorhabdus asymbiotica.
A recently described bangle lectin (PHL) from the bacterium Photorhabdus asymbiotica was identified as a mainly fucose-binding protein that could play an important role in the host-pathogen interaction and in the modulation of host immune response. Structural studies showed that PHL is a homo-dimer that contains up to seven L-fucose-specific binding sites per monomer. For these reasons, potential ligands of the PHL lectin: α-L-fucopyranosyl-containing mono-, di-, tetra-, hexa- and dodecavalent ligands were tested. Two types of polyvalent structures were investigated - calix[4]arenes and dendrimers. The shared feature of all these structures was a C-glycosidic bond instead of the more common but physiologically unstable O-glycosidic bond. The inhibition potential of the tested structures was assessed using different techniques - hemagglutination, surface plasmon resonance, isothermal titration calorimetry, and cell cross-linking. All the ligands proved to be better than free L-fucose. The most active hexavalent dendrimer exhibited affinity three orders of magnitude higher than that of standard L-fucose. To determine the binding mode of some ligands, crystal complex PHL/fucosides 2 - 4 were prepared and studied using X-ray crystallography. The electron density in complexes proved the presence of the compounds in 6 out of 7 fucose-binding sites.


Methodology for the design and economic assessment of anaerobic digestion plants to produce energy and biofertilizer from livestock waste.
The generation and poor disposal of waste from livestock industries is the major cause of pollution of water sources, soil, and air. Therefore, profitable alternatives are required for their correct disposal and use. Anaerobic digestion plants are a technologically viable solution to overcome this problem. In this study, it is proposed a methodology for the design and economic assessment of projects using anaerobic digestion plants to produce electrical energy, thermal energy, and biofertilizer from livestock waste. The methodology is developed based on the assumption that the process is mainly composed of an anaerobic digester and an electric generator having a Diesel-cycle internal combustion engine. It is programmed in "MS Excel" sheet and assessed using technical and economic data from a three real anaerobic digestion plants. The methodology obtains technical parameters such as energy production efficiency with an average difference of 35% compared to the real plants data. In addition, the unit capital costs are calculated, obtaining a value of €3789/kW with a difference of 21.1%, as well as unit operating costs of €729/kW per year with a difference of 15.2%. The financial viability of the project is assessed by calculating the net present value and obtaining €577,050 with a difference of 17.8% and an internal rate of return with a percentage difference of 3%. The proposed methodology specifies the technical parameters and the basic engineering of an anaerobic digestion plant in a stationary state, where the basic streams and dimensions of primary equipment, such as anaerobic reactors and electric generators, are specified. Moreover, the methodology calculates capital and operating expenses for an anaerobic digestion plant, which may be useful to assess the technical and financial feasibility for a project of this type.


[Botulinum toxin in the treatment of pain].
Botulinum toxin (BTx) has been administered for many years in the treatment of dystonias with great success. Its effectiveness is comparable with the best drugs. It was observed during spasmodic torticollis treatment that pain disappears as first before clinical improvement of dystonia. Different mechanisms of influence of BTx on pain are discussed. BTx was tried in tension headache, cluster headache, migraine, fibromyositis, painful cramps with varying results. It is possible that BTx will be useful in many other types of pain.


The Influence of Relative Humidity and Storage Conditions on the Physico-chemical Properties of Inhalation Grade Fine Lactose.
Dry powder inhalers (DPIs) are favorable devices for the delivery of dry formulations to the lungs; still, they largely fail to deliver higher doses of active pharmaceutical ingredient (API) to the lower airways. Addition of fine particles of excipient (fines) to the blend of API and carrier was shown to improve aerosolization performance. Lactose monohydrate is ubiquitous excipient used for this purpose. Lactose exists in a thermodynamically stable crystalline form; however, processes like milling, sieving, or even mixing may induce alteration of crystalline structure and introduce amorphous domains, which could further affect the physico-chemical properties of the material. Therefore, the aim of this work is a detailed characterization of two commercially available types of inhalation grade fine lactose powders (Inhalac 400 and Inhalac 500) prepared using different air-jet milling parameters, with a focus on impact of storage conditions on material properties. We found that the different milling parameters resulted in variable particle size distribution (PSD), and thus surface areas, variable initial amorphous content, cohesivity, flowability, and moisture sorption of materials. In addition, exposure of fine powders to higher humidity reduced the amorphous content present in the materials, but also affected agglomeration tendency and dispersion behavior of both powders. We believe the obtained findings to be important for the aerosolization performance of carrier-based DPIs containing fines and thus need to be duly considered during formulation development.