Axillary metaplastic breast carcinoma with ipsilateral pectoral invasive ductal carcinoma: an unusual presentation.
We report a case of axillary metaplastic breast carcinoma (MBC) with triple negative (ER-/PR-/Her2-) phenotype, concurrent with multifocal invasive ductal carcinoma (IDC) of ipsilateral pectoral breast (ER+/PR+/Her2-) in a 60-year-old woman. The two tumors demonstrate different morphology, immunophenotype, and opposite response to neoadjuvant chemotherapy of paclitaxol, adriamycin, and cyclophosphamide. Methylation analysis of human androgen receptor (HUMARA) on X-chromosome identified monoclonal pattern of X-chromosome inactivation in MBC and mosaic pattern in the IDC. Stem cell origin of MBC is suggested in this case. Clinicopathological features, imaging findings, biological markers, chemoradiation management, and prognosis of MBC are reviewed in comparison to invasive ductal carcinoma. Our case and literature review suggest that traditional chemotherapy applicable to IDC is less effective towards MBC. However, new chemotherapy protocols targeting stem cell and multimodality management of MBC are promising. Recognition of unusual presentation of MBC will help tailor therapy towards tumor with worse prognosis.


Mycobiota in Chilean chilli Capsicum annuum L. used for production of Merkén.
This work aims to provide the first study on the mycobiota present in Chilean pepper Capsicum annuum L. cv. "Cacho de Cabra" throughout the early production stages. Two hundred and forty berry fruits were sampled: 1) at the ripe fruits harvest day; 2) during drying; and 3) smoking processes. A total of 192 strains, encompassing 11 genera and 44 species, were identified through analysis of β-tubulin (benA) gene and internal transcribed spacer of ribosomal DNA (ITS) region. All collection points showed samples with high fungal contamination, but the mycobiota composition varied as a result of different environmental conditions. Alternaria spp. and Fusarium spp. were predominantly isolated from fresh fruits of C. annuum. Penicillium spp. was the most frequent genus in all analysed points. Penicillium brevicompactum and P. crustosum were the most abundant species. Among Aspergillus, A. niger and A. flavus were dominant after the drying phase. In our study, none of the analysed strains of Penicillium (113) and Aspergillus (35) produced Ochratoxin A at detectable levels. The broad characterization of the fungal community of C. annuum carried out in this study, could be a guideline for future mycotoxin analyses performed directly on the pod. Understanding the role and dynamics of mycobiota and its relationship with the toxins present in this substrate, will be useful to establish and improve control measures considering the specificities of each point in the C. annuum production chain.


Basic statistics for clinical pathway evaluation.
Clinical pathways (using accepted benchmark goals) are among the most widespread tools used to enhance outcomes and contain costs within a constrained length of stay (LOS). The author describes the need for selection and use of designated basic statistics in the "definition and evaluation of the impact of clinical pathways in hospitals." The use of both the mean and median data on resource use and LOS information (to evaluate the effectiveness of various clinical pathways) is advised, as the influence of outlier data will thus be revealed. The ultimate goal of such evaluation is the identification of statistics that can be readily "understood by and communicated among providers and consumers of health care services." The correlation of statistical variables such as resource utilization (including ICU stays) and LOS, can reveal patterns not easily seen otherwise.


Biodiversity in the Cladosporium herbarum complex (Davidiellaceae, Capnodiales), with standardisation of methods for Cladosporium taxonomy and diagnostics.
The Cladosporium herbarum complex comprises five species for which Davidiella teleomorphs are known. Cladosporium herbarum s. str. (D. tassiana), C. macrocarpum (D. macrocarpa) and C. bruhnei (D. allicina) are distinguishable by having conidia of different width, and by teleomorph characters. Davidiella variabile is introduced as teleomorph of C. variabile, a homothallic species occurring on Spinacia, and D. macrospora is known to be the teleomorph of C. iridis on Iris spp. The C. herbarum complex combines low molecular distance with a high degree of clonal or inbreeding diversity. Entities differ from each other by multilocus sequence data and by phenetic differences, and thus can be interpreted to represent individual taxa. Isolates of the C. herbarum complex that were formerly associated with opportunistic human infections, cluster with C. bruhnei. Several species are newly described from hypersaline water, namely C. ramotenellum, C. tenellum, C. subinflatum, and C. herbaroides. Cladosporium pseudiridis collected from Iris sp. in New Zealand, is also a member of this species complex and shown to be distinct from C. iridis that occurs on this host elsewhere in the world. A further new species from New Zealand is C. sinuosum on Fuchsia excorticata. Cladosporium antarcticum is newly described from a lichen, Caloplaca regalis, collected in Antarctica, and C. subtilissimum from grape berries in the U.S.A., while the new combination C. ossifragi, the oldest valid name of the Cladosporium known from Narthecium in Europe, is proposed. Standard protocols and media are herewith proposed to facilitate future morphological examination of Cladosporium spp. in culture, and neotypes or epitypes are proposed for all species treated.


Primary Metabolism during Biosynthesis of Secondary Wall Polymers of Protoxylem Vessel Elements.
Xylem vessels, the water-conducting cells in vascular plants, undergo characteristic secondary wall deposition and programmed cell death. These processes are regulated by the VASCULAR-RELATED NAC-DOMAIN (VND) transcription factors. Here, to identify changes in metabolism that occur during protoxylem vessel element differentiation, we subjected tobacco (Nicotiana tabacum) BY-2 suspension culture cells carrying an inducible VND7 system to liquid chromatography-mass spectrometry-based wide-target metabolome analysis and transcriptome analysis. Time-course data for 128 metabolites showed dynamic changes in metabolites related to amino acid biosynthesis. The concentration of glyceraldehyde 3-phosphate, an important intermediate of the glycolysis pathway, immediately decreased in the initial stages of cell differentiation. As cell differentiation progressed, specific amino acids accumulated, including the shikimate-related amino acids and the translocatable nitrogen-rich amino acid arginine. Transcriptome data indicated that cell differentiation involved the active up-regulation of genes encoding the enzymes catalyzing fructose 6-phosphate biosynthesis from glyceraldehyde 3-phosphate, phosphoenolpyruvate biosynthesis from oxaloacetate, and phenylalanine biosynthesis, which includes shikimate pathway enzymes. Concomitantly, active changes in the amount of fructose 6-phosphate and phosphoenolpyruvate were detected during cell differentiation. Taken together, our results show that protoxylem vessel element differentiation is associated with changes in primary metabolism, which could facilitate the production of polysaccharides and lignin monomers and, thus, promote the formation of the secondary cell wall. Also, these metabolic shifts correlate with the active transcriptional regulation of specific enzyme genes. Therefore, our observations indicate that primary metabolism is actively regulated during protoxylem vessel element differentiation to alter the cell's metabolic activity for the biosynthesis of secondary wall polymers.


Classifying the courses of schizophrenia.
The purpose of this review was to examine current strategies for classifying the courses of schizophrenia. Although a number of course classifications have evolved over the past two decades, the field has not addressed fully the complex issue of course description. This review compared previously published prototypes for describing the long-term courses of schizophrenia to identify variations and consistencies in course descriptions. Across investigations, differences were found in the number of courses described, in the structure of course categories, in the relative emphasis on syndrome symptoms versus syndrome change, and in documenting specific course features (i.e., illness onset, illness outcome, and types of symptoms). Interstudy comparisons of long-term illness patterns therefore are limited and cumulative statements on prognosis will remain problematic until some standardization is introduced. A minimal set of course parameters that should be documented in future studies include (1) the rate of syndrome onset; (2) postonset patterns of psychotic and residual symptoms; (3) postonset patterns of social, work, and self-care activities; and (4) outcome. Descriptive domains for assessing these parameters are offered. Future research should be directed toward (1) developing a standard schizophrenia course classification that is more representative than the current DSM and Research Diagnostic Criteria classifications; (2) using a broadened domain of symptom patterns to characterize short- and long-term courses of schizophrenia; (3) increasing course reports with a focus on patients who demonstrate intermediate illness courses; and (4) exploring factors that converge with particular course types.


Twenty-Five Novel Loci for Carotid Intima-Media Thickness: A Genome-Wide Association Study in >45 000 Individuals and Meta-Analysis of >100 000 Individuals.
Carotid artery intima-media thickness (cIMT) is a widely accepted marker of subclinical atherosclerosis. Twenty susceptibility loci for cIMT were previously identified and the identification of additional susceptibility loci furthers our knowledge on the genetic architecture underlying atherosclerosis. We performed 3 genome-wide association studies in 45 185 participants from the UK Biobank study who underwent cIMT measurements and had data on minimum, mean, and maximum thickness. We replicated 15 known loci and identified 20 novel loci associated with cIMT at P<5×10-8. Seven novel loci (ZNF385D, ADAMTS9, EDNRA, HAND2, MYOCD, ITCH/EDEM2/MMP24, and MRTFA) were identified in all 3 phenotypes. An additional new locus (LOXL1) was identified in the meta-analysis of the 3 phenotypes. Sex interaction analysis revealed sex differences in 7 loci including a novel locus (SYNE3) in males. Meta-analysis of UK Biobank data with a previous meta-analysis led to identification of three novel loci (APOB, FIP1L1, and LOXL4). Transcriptome-wide association analyses implicated additional genes ARHGAP42, NDRG4, and KANK2. Gene set analysis showed an enrichment in extracellular organization and the PDGF (platelet-derived growth factor) signaling pathway. We found positive genetic correlations of cIMT with coronary artery disease rg=0.21 (P=1.4×10-7), peripheral artery disease rg=0.45 (P=5.3×10-5), and systolic blood pressure rg=0.30 (P=4.0×10-18). A negative genetic correlation between average of maximum cIMT and high-density lipoprotein was found rg=-0.12 (P=7.0×10-4). Genome-wide association meta-analyses in >100 000 individuals identified 25 novel loci associated with cIMT providing insights into genes and tissue-specific regulatory mechanisms of proatherosclerotic processes. We found evidence for shared biological mechanisms with cardiovascular diseases.


Replicon sizes in mammalian cells as estimated by an x-ray plus bromodeoxyuridine photolysis method.
A new method is described for estimating replicon sizes in mammalian cells. Cultures were pulse labeled with [3H]thymidine ([3H]TdR) and bromodeoxyuridine (BrdUrd) for up to 1 h. The lengths of the resulting labeled regions of DNA, Lobs, were estimated by a technique wherein the change in molecular weight of nascent DNA strands, induced by 313 nm light, is measured by velocity sedimentation in alkaline sucrose gradients. If cells are exposed to 1,000 rads of X-rays immediately before pulse labeling, initiation of replicon operation is blocked, although chain elongation proceeds almost normally. Under these conditions Lobs continues to increase only until operating replicons have completed their replication. This value for Lobs then remains constant as long as the block to initiation remains and represents an estimate for the average size of replicons operating in the cells before X-irradiation. For human diploid fibroblasts and human HeLa cells this estimated average size is approximately 17 micron, whereas for Chinese hamster ovary cells, the average replicon size is about 42 micron.


Rectangular coordination polymer nanoplates: large-scale, rapid synthesis and their application as a fluorescent sensing platform for DNA detection.
In this paper, we report on the large-scale, rapid synthesis of uniform rectangular coordination polymer nanoplates (RCPNs) assembled from Cu(II) and 4,4'-bipyridine for the first time. We further demonstrate that such RCPNs can be used as a very effective fluorescent sensing platform for multiple DNA detection with a detection limit as low as 30 pM and a high selectivity down to single-base mismatch. The DNA detection is accomplished by the following two steps: (1) RCPN binds dye-labeled single-stranded DNA (ssDNA) probe, which brings dye and RCPN into close proximity, leading to fluorescence quenching; (2) Specific hybridization of the probe with its target generates a double-stranded DNA (dsDNA) which detaches from RCPN, leading to fluorescence recovery. It suggests that this sensing system can well discriminate complementary and mismatched DNA sequences. The exact mechanism of fluorescence quenching involved is elucidated experimentally and its use in a human blood serum system is also demonstrated successfully.


[Recurrent gastric volvulus resolved surgically after therapeutic percutaneous endoscopic gastrostomy in Duchenne muscular dystrophy].
A 20-year-old man with Duchenne muscular dystrophy (DMD) with recurrent gastric volvulus underwent percutaneous endoscopic gastrostomy (PEG). Four months later, he developed vomiting and consciousness disturbance. CT revealed gastric volvulus recurrence along the gastrostomy axis. Endoscopic repositioning failed and fistula perforation necessitated emergency surgery. The upper position of the stomach was twisted counter-clockwise and revolved on the gastrostomy axis sliding between the lower stomach and abdominal wall. The fistula showed necrotic perforation and was thus resected. The anterior stomach wall was fixed to the abdominal wall at 3 triangular points. Thereafter, gastric volvulus did not recur. PEG is reportedly effective for preventing gastric volvulus, but there are rare cases of postgastrostomy recurrence. This successfully managed case provides valuable clinical insights.


Full implementation of computerized physician order entry and medication-related quality outcomes: a study of 3364 hospitals.
This study compares quality of care measures for hospitals with fully implemented computerized physician order entry (CPOE) systems with hospitals that have not fully implemented such a system. Using a cross-sectional design, this study linked hospital quality data from the Centers for Medicare and Medicaid Services to the Health Information Management Systems Society Analytics database, which contains hospital CPOE adoption information. Performance on quality measures was assessed using univariate and multivariate methods. In all, 8% of hospitals have fully implemented CPOE systems; CPOE hospitals were more frequently larger, not-for-profit, and teaching hospitals. After controlling for confounders, CPOE hospitals outperformed comparison hospitals on 5 of 11 measures related to ordering medications and on 1 of 9 nonmedication-related quality measures. Using a large sample of hospitals, our study found significant positive associations between specific objective quality indicators and CPOE implementation.


Unilateral cleft lip repair by rotation/advancement: potential errors and how to avoid them.
The surgical repair of unilateral cleft lip has been refined over the centuries. During the past few decades, two techniques have emerged as ideal: (1) the rotation/advancement technique and (2) the triangular flap repair, the former more commonly utilized. Underrotation and insufficient advancement represent two potential errors when using this technique. In this review, these errors in flap development will be examined and strategies discussed for their avoidance. Insufficiencies in both rotation and advancement produce functionally and aesthetically suboptimal results. Although the principles of rotation and advancement are well described, differences in opinions exist as to the precise details of flap development. The extent of rotation and the paring point (Millard point #8) of the advancement flap will specifically be addressed. It stands to reason that the severity of the cleft deformity will determine the required surgical intervention. By appropriately applying the techniques herein, satisfactory results can be achieved in even severe clefts.


Spiral CT scanning of the paranasal sinuses.
Because of the interest in reducing the time and radiation required for screening sinuses with CT, we compared the quality of conventional coronal CT images with coronally reconstructed, axially acquired spiral CT images. The spiral reconstructions compared favorably with the conventional images and had significantly less motion and dental amalgam artifacts.


Reformatted images improve the detection rate of acute traumatic subdural hematomas on brain CT compared with axial images alone.
Subdural hematomas (SDHs) comprise a significant percentage of missed intracranial hemorrhage on axial brain CT. SDH detection rates could be improved with the addition of reformatted images. Though performed at some centers, the potential additional diagnostic sensitivity of reformatted images has not yet been investigated. The purpose of our study is to determine if the addition of coronal and sagittal reformatted images to an axial brain CT increases the sensitivity and specificity for detection of acute traumatic SDH. We retrospectively reviewed consecutive brain CTs acquired for acute trauma that contained new SDHs. An equivalent number of normal brain CTs served as control. Paired sets of images were created for each case: (1) axial images only ("axial only") and (2) axial, coronal, sagittal images ("reformat added"). Three readers interpreted both the axial only and companion reformat added for each case, separated by 1 month. Reading times and SDH detection rates were compared. One hundred SDH and 100 negative examinations were collected. Sensitivity and specificity for the axial-only scans were 75.7 and 94.3 %, respectively, compared with 88.3 and 98.3 % for reformat added. There was a 24.3 % false negative (missed SDH) rate with axial-only scans versus 11.7 % with reformat added (p = <0.001). Median reader interpretation times were longer with the addition of reformatted images (125 versus 89 s), but this difference was not significant (p = 0.23). The addition of coronal and sagittal images in trauma brain CT resulted in improved sensitivity and specificity as well as a reduction in SDH false negatives by greater than 50 %. Reformatted images substantially reduce the number of missed SDHs compared with axial images alone.


The effectiveness of exercise-based rehabilitation to patients with myeloproliferative neoplasms-An explorative study.
The aim of the study was to determine the impact of an interdisciplinary exercise-based rehabilitation intervention on fatigue and quality of life (QOL) in patients with Philadelphia-negative myeloproliferative neoplasms (MPNs). At the Danish Knowledge Centre for Rehabilitation and Palliative Care, a 5-day interdisciplinary exercise-based rehabilitation intervention was carried out on 48 patients with MPN. It was followed by 12 weeks of self-exercising prior to follow-up. Initially and at follow-up, participants filled out validated questionnaires; Brief Fatigue Inventory, Multidimensional Fatigue Inventory, European Organization for Research and Treatment of Cancer Quality-of-Life questionnaire, Myeloproliferative Neoplasm Symptom Assessment Form and Hospital Anxiety and Depression Scale. Maximal oxygen uptake and muscular strength were tested as well. Paired t test was used to compare scores between baseline and follow-up. In total, 45 participants (94%) completed the follow-up. No significant differences were observed on fatigue or QOL when comparing baseline and follow-up. Mean maximal oxygen uptake increased from 27.2 to 33.6 ml O2 · kg-1 ·min-1 (p < 0.001). Handgrip strength (p = 0.01) and the 30-s chair-stand test (p < 0.001) improved as well. No changes were found regarding levels of fatigue and QOL. However, we observed a significant increase in the physical capacity. Our observations call for further studies investigating the effects of non-pharmacological approaches in patients with MPN.


The Genetic Management Clinic: Oncology Nurses and Management of Hereditary Cancer Risk.
Integrating clinics designed to manage hereditary cancer risk into oncology care models has the potential to improve navigation of individuals predisposed to increased cancer risk through risk-reducing education, as well as recommendations for cancer screening and surveillance. Oncology nurses have the unique opportunity and optimal position to navigate patients through the complex subject of genetics and hereditary cancer syndromes. Effectively and efficiently navigating patients with pathogenic variants to minimize and manage hereditary cancer risk has the potential to improve patient outcomes.


Identification of impurities in erythromycin by liquid chromatography-mass spectrometric detection.
A simple, isocratic liquid chromatographic (LC) method using volatile mobile phase constituents for the identification of related substances in erythromycin samples is described. For method development, evaporative light scattering detection (ELSD) was used. An XTerra RP18 column was used at 70 degrees C with a mobile phase consisting of acetonitrile-isopropanol-0.2M ammonium acetate pH 7.0-water (165:105:50:680). Mass spectral data were acquired on an ion trap mass spectrometer equipped with an electrospray interface operated in the positive ion mode. First, a library was created using MS/MS and MS(n) spectra of reference substances available in the laboratory. Using these reference spectra as interpretative templates, eight novel related substances in erythromycin samples were identified: N-demethylerythromycin E, erythromycin E N-oxide, anhydroerythromycin C, N-demethylerythromycin B, anhydro-N-demethylerythromycin A, pseudoerythromycin E enol ether, EF lacking the neutral sugar and EA lacking the neutral sugar.


Allosteric modulation of myristate and Mn(III)heme binding to human serum albumin. Optical and NMR spectroscopy characterization.
Human serum albumin (HSA) is best known for its extraordinary ligand binding capacity. HSA has a high affinity for heme and is responsible for the transport of medium and long chain fatty acids. Here, we report myristate binding to the N and B conformational states of Mn(III)heme-HSA (i.e. at pH 7.0 and 10.0, respectively) as investigated by optical absorbance and NMR spectroscopy. At pH 7.0, Mn(III)heme binds to HSA with lower affinity than Fe(III)heme, and displays a water molecule coordinated to the metal. Myristate binding to a secondary site FAx, allosterically coupled to the heme site, not only increases optical absorbance of Mn(III)heme-bound HSA by a factor of approximately three, but also increases the Mn(III)heme affinity for the fatty acid binding site FA1 by 10-500-fold. Cooperative binding appears to occur at FAx and accessory myristate binding sites. The conformational changes of the Mn(III)heme-HSA tertiary structure allosterically induced by myristate are associated with a noticeable change in both optical absorbance and NMR spectroscopic properties of Mn(III)heme-HSA, allowing the Mn(III)-coordinated water molecule to exchange with the solvent bulk. At pH = 10.0 both myristate affinity for FAx and allosteric modulation of FA1 are reduced, whereas cooperation of accessory sites and FAx is almost unaffected. Moreover, Mn(III)heme binds to HSA with higher affinity than at pH 7.0 even in the absence of myristate, and the metal-coordinated water molecule is displaced. As a whole, these results suggest that FA binding promotes conformational changes reminiscent of N to B state HSA transition, and appear of general significance for a deeper understanding of the allosteric modulation of ligand binding properties of HSA.


Education, Knowledge, and Practice Characteristics of Cannabis Physicians: A Survey of the Society of Cannabis Clinicians.
Context: Medical cannabis use has increased in recent years despite being a federally illegal drug in the United States. States with medical cannabis use laws require patients to be certified by physicians. However, little is known about the education, knowledge, and practice characteristics of physicians who recommend and supervise patients' use of medical cannabis. Objective: This study assessed how U.S. physicians who practice cannabis medicine are educated, self-assess their knowledge, and describe their practice. Methods: In fall 2017, a 57-item, electronic survey was sent to all members of the Society of Cannabis Clinicians. Because California has had legalized medical cannabis for longer than any other state, we analyzed responses for 14 items between California and non-California physicians. Results: Of 282 surveyed, 133 were eligible and 45 completed the survey. Of those, multiple medical specialties were represented. Only one physician received education during medical school about cannabis medicine, but physicians gained knowledge through conferences (71%, 32/45), the medical literature (64%, 29/45), and websites (62%, 28/45). Just over half (56%, 20/45) felt that there was sufficient information available to practice cannabis medicine. Of the 37 who answered the knowledge question, most felt knowledgable about cannabinoids (78%, 29/37) and the endocannabinoid system (76%, 28/37). There was a wide variation in the number of cannabis recommendations provided by physicians over the course of their practice career (median 1200; interquartile range, 100-5000), and most provided condition-specific treatment (69%, 31/45) and dosing recommendations (62%, 28/45). The majority (81%, 30/37) of physicians received referrals from mainstream medical providers. No differences were found between California and non-California physicians, except more women were from California (p=0.02). Conclusions: The use of medical cannabis continues to increase in the United States and globally. All states that allow medical cannabis require a physician's recommendation, yet few states require specific clinical training. Findings of this study suggest the need for more formal education and training of physicians in medical school and residency, more opportunities for cannabis-related continuing medical education for practicing physicians, and clinical and basic science research that will inform best practices in cannabis medicine.


[Ecological aggression and adaptation mechanisms].
The article presents the main ecological hazards and human adaptational mechanisms. Special attention is paid to chemicals being highly prevalent in environment and to mechanisms of their activation and deactivation by means of enzymes dependent on cytochrome P450.


The Effect of Age of Initiation of Cannabis Use on Psychosis, Depression, and Anxiety among Youth under 25 Years.
This study was conducted to review the current state of evidence on the association between age of initiation of cannabis use and symptoms of psychosis, depression, or anxiety among youth under 25 years of age. We conducted a systematic review of articles published prior to March 2018 by searching OVID MEDLINE, PsycINFO, EMBASE, and the references of included studies. We included comparative studies (cohort, case-control, cross-sectional) that reported on cannabis use in persons <25 years of age (exposure) and symptoms of psychosis, depression, or anxiety (outcome). We narratively synthesized the studies according to design (cohort, etc.) and psychiatric outcome. We used the Newcastle-Ottawa Scale to assess risk of bias. Of the 534 citations identified through the literature search, 23 met the eligibility criteria and were included in this review. With psychosis as the outcome, all except one study found that earlier cannabis use was generally associated with higher risks. With depression/anxiety as the outcome, 6 of the 11 included studies reported findings indicating that earlier use of cannabis was linked to higher symptom levels. In persons <25 years old, greater cannabis use is associated with more psychological symptoms, especially among those with a predisposition or existing vulnerability to such outcomes (Oxford Centre for Evidence-Based Medicine level 3 or 4). Policy makers need to consider the adverse effects of cannabis use in youth when planning a public health approach to cannabis legalization.


Pooled-DNA sequencing identifies novel causative variants in PSEN1, GRN and MAPT in a clinical early-onset and familial Alzheimer's disease Ibero-American cohort.
Some familial Alzheimer's disease (AD) cases are caused by rare and highly-penetrant mutations in APP, PSEN1, and PSEN2. Mutations in GRN and MAPT, two genes associated with frontotemporal dementia (FTD), have been found in clinically diagnosed AD cases. Due to the dramatic developments in next-generation sequencing (NGS), high-throughput sequencing of targeted genomic regions of the human genome in many individuals in a single run is now cheap and feasible. Recent findings favor the rare variant-common disease hypothesis by which the combination effects of rare variants could explain a large proportion of the heritability. We utilized NGS to identify rare and pathogenic variants in APP, PSEN1, PSEN2, GRN, and MAPT in an Ibero-American cohort. We performed pooled-DNA sequencing of each exon and flanking sequences in APP, PSEN1, PSEN2, MAPT and GRN in 167 clinical and 5 autopsy-confirmed AD cases (15 familial early-onset, 136 sporadic early-onset and 16 familial late-onset) from Spain and Uruguay using NGS. Follow-up genotyping was used to validate variants. After genotyping additional controls, we performed segregation and functional analyses to determine the pathogenicity of validated variants. We identified a novel G to T transition (g.38816G>T) in exon 6 of PSEN1 in a sporadic early-onset AD case, resulting in a previously described pathogenic p.L173F mutation. A pathogenic p.L392V mutation in exon 11 was found in one familial early-onset AD case. We also identified a novel CC insertion (g.10974_10975insCC) in exon 8 of GRN, which introduced a premature stop codon, resulting in nonsense-mediated mRNA decay. This GRN mutation was associated with lower GRN plasma levels, as previously reported for other GRN pathogenic mutations. We found two variants in MAPT (p.A152T, p.S318L) present only in three AD cases but not controls, suggesting that these variants could be risk factors for the disease. We found pathogenic mutations in PSEN1, GRN and MAPT in 2.33% of the screened cases. This study suggests that pathogenic mutations or risk variants in MAPT and in GRN are as frequent in clinical AD cases as mutations in APP, PSEN1 and PSEN2, highlighting that pleiotropy of MAPT or GRN mutations can influence both FTD and AD phenotypic traits.


Regulation of 25-hydroxyvitamin D3 metabolism in chick embryo.
We investigated the time course of the development of renal 25-hydroxyvitamin D-1-hydroxylase and 25-hydroxyvitamin D-24-hydroxylase in chick embryos grown in the presence and absence of the eggshell. In embryos with the eggshell, the specific activity (SA) of 25-hydroxyvitamin D-1-hydroxylase in kidney homogenates increased from 0.68 fmol X min-1 X mg protein-1 at 12 days of gestation to a peak of 2.55 +/- 0.50 fmol X min-1 X mg-1 protein-1 at 17 days. In contrast, the SA of 25-hydroxyvitamin D-24-hydroxylase decreased from 2.5 fmol X min-1 X mg protein-1 to 0.90 +/- 0.25 fmol X min-1 X mg protein-1 during the interval. The total plasma calcium was significantly reduced in embryos without shells at 14 to 15 days of gestation (1.1 +/- 0.1 mM, mean +/- SE) compared with normal embryos of the same gestation (2.3 +/- 0.3 mM, P less than 0.002). In embryos without the eggshell, renal 25-hydroxyvitamin D-1-hydroxylase increased from 6.0 to 8.2 +/- 0.6 fmol X min-1 X mg protein-1 at 17 days of gestation and was from four- to sixfold higher than corresponding enzymatic activities for intact embryos. The increased enzyme activity resulting from loss of the eggshell was due to an increase in Vmax. The findings indicate that renal 25-hydroxyvitamin D-1-hydroxylase and 25-hydroxyvitamin D-24-hydroxylase in the chick embryo exhibit activity and show a large capacity for regulation in response to changes in calcium metabolism.


The role of negative costimulators during parasitic infections.
Activation of T lymphocytes through TCR signalling takes place within the context of numerous other cell surface proteins. To prevent unnecessary activation of T cells the immune system has developed an intricate balance between positive and negative costimulatory signals. Positive costimulatory signals determine whether antigen recognition by T cells leads to full activation or to anergy/death. In contrast, the expression of negative costimulatory molecules by T cells such as cytotoxic T lymphocyte antigen 4 (CTLA-4) mediates the regulation of an immune response and thus plays a pivotal role in the maintenance of peripheral tolerance. The new members of the CD28 family members, programmed death-1 (PD-1) and B and T lymphocyte attenuator (BTLA) are expressed more broadly on various immune cells and are also induced upon inflammation. There is increasing evidence that negative costimulators are critically involved in the regulation of immune responses against parasitic infections. The production of pro-inflammatory cytokines is often required to control parasites but the same cytokines contribute to immunopathology. Recent studies indicate that these negative costimulators are not redundant but fulfil specific functions in different tissues. This might represent a means to provoke a fine-tuning of the immune response and to define tissue specific limits of inflammation that ensure proper physiological function of the respective organ. Thus, negative costimulators represent possible drug targets since their blockade leads to an increased immune response whereas their ligation dampens T cell activation and thereby might prevent immunopathology. This review examines the role of negative costimulators during parasitic infections, and we also discuss their therapeutic potential.


Emergent radiotherapy for brain and leptomeningeal metastases: a narrative review.
As novel systemic therapies allow patients to live longer with cancer, the risk of developing central nervous system (CNS) metastases increases and providers will more frequently encounter emergent presentation of brain metastases (BM) and leptomeningeal metastases (LM). Management of these metastases requires appropriate work-up and well-coordinated multidisciplinary care. We set out to perform a review of emergent radiotherapy (RT) for CNS metastases, specifically focusing on BM and LM. We review the appropriate pathways for workup and initial management of BM and LM, while reviewing the literature supporting emergent treatment of these entities with surgery, systemic anti-cancer therapy, and RT. To inform this narrative review, literature searches in PubMed and Google Scholar were conducted, with preference given to articles employing modern RT techniques, when applicable. Due to the paucity of high-quality evidence for management of BM and LM in the emergent setting, discussion was supplemented by the authors' expert commentary. This work highlights the importance of surgical evaluation, particularly for patients presenting with significant mass effect, hemorrhagic metastases, or increased intracranial pressure. We review the rare situations where emergent initiation of systemic anti-cancer therapy is indicated. When defining the role of RT, we review factors guiding selection of appropriate modality, treatment volume, and dose-fractionation. Generally, 2D- or 3D-conformal treatment techniques prescribed as 30 Gy in 10 fractions or 20 Gy in 5 fractions, should be employed in the emergent setting. Patients with BM and LM present from a diverse array of clinical situations, requiring well-coordinated multidisciplinary management, and there is a paucity of high-quality evidence guiding such management decisions. This narrative review aims to more thoroughly prepare providers for the challenging situation of emergent management of BM and LM.


Schisantherin A alleviates non-alcoholic fatty liver disease by restoring intestinal barrier function.
Non-alcoholic fatty liver disease (NAFLD) is intricately linked to dysregulation of the gut-liver axis, and correlated with intestinal inflammation and barrier disruption. To investigate the protective effects and possible molecular mechanism of Schisantherin A (Sin A) in a high-fat diet (HFD) induced NAFLD mouse model. HFD-fed NAFLD mice were treated with the vehicle and 80 mg/kg Sin A every day for 6 weeks. The gut permeability of the NAFLD mice was assessed by intestinal permeability assays in vivo and transepithelial electrical resistance (TEER) measurements in vitro were also used to evaluate the function of the gut barrier. TLR4 inhibitor was then used to investigate the impact of Sin A in the LPS- TLR4 signaling pathway. Alternatively, the composition of the microbiome was assessed using 16S rRNA amplification. Finally, the experiment of antibiotic treatment was performed to elucidate the roles of the gut microbiome mediating Sin A induced metabolic benefits in the NAFLD mice. We found that Sin A potently ameliorated HFD-induced hepatic steatosis and inflammation, alleviated gut inflammation, and restored intestinal barrier function. We also observed that Sin A improved gut permeability and reduced the release of lipopolysaccharide (LPS) into circulation and further found that Sin A can suppress LPS-TLR4 signaling to protect against HFD-induced NAFLD. Sin A treatment altered the composition of the microbiome in NAFLD mice compared to vehicle controls. Sin A is an effective and safe hepatoprotective agent against HFD-induced NAFLD by partly ameliorating gut inflammation, restoring intestinal barrier function, and regulating intestinal microbiota composition.


What is the percentage of pathogenic borreliae in spirochaetal findings of mosquito larvae?
In this study, larvae (1,179 ex.) of mosquito genera Culex were examined for the presence of spirochaetes by Dark Field Microscopy (DFM) at the locality of Blansko (Czech Republic) in of 2004-2008. DFM spirochaete positive samples (25.4 %) were investigated by nested PCR; only 4 samples were positive for the presence of Borrelia burgdorferi sensu lato, which is 0.3 % of the total examined samples. We can conclude that only a low percentage of pathogenic borreliae are presented in mosquito larvae, while the spirochaete of undefined genera infect larvae in high amounts.


Nurses' Information, Attıtude and Practices towards Use of Physical Restraint in Intensive Care Units.
Introduction: Physical restraint may seem to be a useful and simple procedure to help the treatment but is a complex practice including physical, psychological, judicial, ethical and moral issues. Research was made on description basis in order to determine the knowledge, attitude and application levels of nurses working in critical care units about physical restraint applied on patients. Methods: The study was performed as a descriptive and correlation study. Working in ICUs, 158 nurses constituted the sampling. "Levels of Knowledge, Attitudes and Practices of Staff Regarding Physical Restraints Questionnaire" was used to collect data. Results: For information, attitude and practice scores, participants' scores were 7.1(1.7), 31.8 (4.6) and 36.6 (3.2), respectively. No association was found between information subscale, and age, professional years, working time in ICUs and weekly working hours. However, for attitude subscale, a negative and weak association was found between age (r=-0.229) and professional years (r=-0.174), and increasing these variables decreased attitude score. No association was found between attitude score, and working time in ICUs and weekly working hours. While there was no association between practice score, and age, professional years and working time in ICUs, the increase in weekly working hours (r=-0.243) was found to decrease practice score, and this association was found weak. Conclusion: In conclusion, we consider nurses' level of information is sufficient, but attitudes and practice were not at a positive level. It is recommendable that out of such conditions, novel approaches should be developed to decrease the use of physical restraint.


Novel solid-state polymer electrolyte consisting of a porous layer-by-layer polyelectrolyte thin film and oligoethylene glycol.
A novel solid-state polymer electrolyte was constructed using layer-by-layer (LbL) polyelectrolyte assembly of linear poly(ethylenimine) (LPEI) and poly(acrylic acid) (PAA), combined with a plasticization step using oligoethylene glycol dicarboxylic acid (OEGDA). This composite film exhibits a relatively high ionic conductivity of 9.5 x 10(-5) S/cm at 25 degrees C and 22% relative humidity. Detailed characterization of the composite was undertaken using grazing-angle Fourier transform infrared (GA-FTIR), atomic force microscopy (AFM), scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and impedance spectroscopy. After immersing the LPEI/PAA films into OEGDA aqueous solutions, the films exhibited a swelling behavior and increased surface roughness indicative of porosity induced by reorganization of ionic interactions between LPEI and PAA in acidic solution. This internal porous structure allows inclusion of OEGDA within the multilayer and increased ionic conductivity under ambient conditions due to the combined effects of plasticization of the LbL matrix by atmospheric water as well as the added mobility of ions in molten OEGDA within the composite.


Creating the medicine of tomorrow: ten hot Hopkins research areas today.
From its very beginnings, the Johns Hopkins School of Medicine has fostered the development of physician scientists whose research and practice of medicine are closely intertwined. As a result, the long list of medical breakthroughs--of Hopkins "firsts"--has put the institution at the forefront of medical research in the past century. This tradition continues today, with research in many and diverse areas. Ten current projects underway at Hopkins, attempting to unravel mysteries in neurology, oncology, genetics, hematology, and nephrology, are described in this article. All hold promise to continue the reputation of Johns Hopkins as a leader in research, to become new milestones in the progress of modern medicine.


LAMB3 Missense Variant in Australian Shepherd Dogs with Junctional Epidermolysis Bullosa.
In a highly inbred Australian Shepherd litter, three of the five puppies developed widespread ulcers of the skin, footpads, and oral mucosa within the first weeks of life. Histopathological examinations demonstrated clefting of the epidermis from the underlying dermis within or just below the basement membrane, which led to a tentative diagnosis of junctional epidermolysis bullosa (JEB) with autosomal recessive inheritance. Endoscopy in one affected dog also demonstrated separation between the epithelium and underlying tissue in the gastrointestinal tract. As a result of the severity of the clinical signs, all three dogs had to be euthanized. We sequenced the genome of one affected puppy and compared the data to 73 control genomes. A search for private variants in 37 known candidate genes for skin fragility phenotypes revealed a single protein-changing variant, LAMB3:c.1174T>C, or p.Cys392Arg. The variant was predicted to change a conserved cysteine in the laminin β3 subunit of the heterotrimeric laminin-322, which mediates the binding of the epidermal basement membrane to the underlying dermis. Loss-of-function variants in the human LAMB3 gene lead to recessive forms of JEB. We confirmed the expected co-segregation of the genotypes in the Australian Shepherd family. The mutant allele was homozygous in two genotyped cases and heterozygous in three non-affected close relatives. It was not found in 242 other controls from the Australian Shepherd breed, nor in more than 600 other controls. These data suggest that LAMB3:c.1174T>C represents the causative variant. To the best of our knowledge, this study represents the first report of a LAMB3-related JEB in domestic animals.


Genome Wide Analysis of the Transcriptional Profiles in Different Regions of the Developing Rice Grains.
Rice is an important food source for humans worldwide. Because of its nutritional and agricultural significance, a number of studies addressed various aspects of rice grain development and grain filling. Nevertheless, the molecular processes underlying grain filling and development, and in particular the contributions of different grain tissues to these processes, are not understood. Using RNA-sequencing, we profiled gene expression activity in grain tissues comprised of cross cells (CC), the nucellar epidermis (NE), ovular vascular trace (OVT), endosperm (EN) and the aleurone layer (AL). These tissues were dissected using laser capture microdissection (LCM) at three distinct grain development stages. The mRNA expression datasets offer comprehensive and new insights into the gene expression patterns in different rice grain tissues and their contributions to grain development. Comparative analysis of the different tissues revealed their similar and/or unique functions, as well as the spatio-temporal regulation of common and tissue-specific genes. The expression patterns of genes encoding hormones and transporters indicate an important role of the OVT tissue in metabolite transport during grain development. Gene co-expression network prediction on OVT-specific genes identified several distinct and common development-specific transcription factors. Further analysis of enriched DNA sequence motifs proximal to OVT-specific genes revealed known and novel DNA sequence motifs relevant to rice grain development. Together, the dataset of gene expression in rice grain tissues is a novel and useful resource for further work to dissect the molecular and metabolic processes during rice grain development.


Efficacy and safety of alirocumab, a monoclonal antibody to PCSK9, in statin-intolerant patients: design and rationale of ODYSSEY ALTERNATIVE, a randomized phase 3 trial.
Statin intolerance has been a major limitation in the use of statins, especially at higher doses. New effective treatments are needed for lowering low-density lipoprotein cholesterol (LDL-C) in patients who cannot tolerate daily statin doses. ODYSSEY ALTERNATIVE (NCT01709513) evaluates efficacy and safety of alirocumab, a fully human proprotein convertase subtilisin/kexin type 9 monoclonal antibody, in patients with well-documented statin intolerance and moderate to very high cardiovascular risk. This is a phase 3, multicenter, randomized, double-blind, double-dummy study in statin-intolerant patients. Intolerance was defined as inability to take at least 2 different statins because of muscle-related adverse events (AEs), 1 at the lowest approved starting dose. Patients first received single-blind subcutaneous and oral placebo for 4 weeks, and were withdrawn if they developed muscle-related AEs after the placebo treatment. Continuing patients were randomized (2:2:1 ratio) to alirocumab 75 mg self-administered via single 1 mL prefilled pen every 2 weeks or ezetimibe 10 mg/day or atorvastatin 20 mg/day (statin rechallenge), for 24 weeks. Alirocumab dose was increased to 150 mg every 2 weeks (also 1 mL) at week 12 depending on week 8 LDL-C level. The primary endpoint is percent change in LDL-C from baseline to week 24 by intent-to-treat analysis. Muscle-related AEs were assessed by spontaneous patient reports and clinic queries. A total of 314 patients have been randomized. This is the first and only study of a new class of LDL-C-lowering agents in patients selected with a rigorously documented intolerance to statins, using a placebo run-in and statin control arm.


Classification of opioids on the basis of change in seizure threshold in rats.
Twenty opioids have been subdivided into four classes by using flurothyl-induced seizures in rats to measure dose-response relationships, stereospecificity, naloxone sensitivity, and tolerance-cross-tolerance. The data support current theories of multiple opiate receptor types. Since the receptors involved mediate effects that are antagonized, enhanced, or unaffected by naloxone, the model is uniquely suitable for detecting novel narcotic antagonists that can then be used to differentiate opiate receptors in other systems.


Effect of indacaterol on lung deflation improves cardiac performance in hyperinflated COPD patients: an interventional, randomized, double-blind clinical trial.
COPD is often associated with cardiovascular comorbidity. Treatment guidelines recommend therapy with bronchodilators as first choice. We investigated the acute effect of single-dose indacaterol on lung hyperinflation in COPD subjects, for the first time evaluating the potential effects on right heart performance. In this Phase IV, randomized, interventional, double-blind, crossover clinical study, we recruited 40 patients (50-85 years of age) with stable COPD. Patients were treated with 150 μg indacaterol or placebo and after 60 minutes (T60) and 180 minutes (T180) the following tests were performed: trans-thoracic echocardiography (TTE), plethysmography, diffusing capacity of the lung for carbon monoxide, saturation of peripheral oxygen, and visual analog scale dyspnea score. Patients underwent a crossover re-challenge after a further 72 hours of pharmacological washout. All TTE measurements were conducted blindly by the same operator and further interpreted by two different blinded operators. Consensus decisions were taken on every value and parameter. The primary outcome was the effect of the reduction of residual volume and functional residual capacity on right heart systolic and diastolic function indexes evaluated by TTE in patients treated with indacaterol, as compared to placebo. Vital capacity, inspiratory capacity, and forced expiratory volume in 1 second were significantly increased by indacaterol, when compared with placebo, while residual volume, intrathoracic gas volume, and specific airway resistance were significantly reduced in patients treated with indacaterol. Tricuspid annular plane systolic excursion was significantly increased versus placebo, paralleled by an increase of tricuspid E-wave deceleration time. The cardiac frequency was also significantly reduced in indacaterol-treated patients. Indacaterol significantly reduces lung hyperinflation in acute conditions, with a clinically relevant improvement of dyspnea. These modifications are associated with a significant increase of the right ventricular compliance indexes and may have a role in improving left ventricular preload leading to a reduction in cardiac frequency.


[Autoimmune encephalitis. A review].
Autoimmune encephalitis are one of the emergent causes of subacute changes in the level of consciousness, behavior, cognitive impairment and seizures, mainly in young people. They are a consequence of inflammation or dysfunction of parts of the brain caused by antibodies against specific brain antigens, usually located in the limbic system, resulting in clinical presentation as a limbic encephalitis. The objectives of this article are to show the clinical presentation, complementary studies and treatment of this entity, considering that the patient's prognostic depends on a high level of clinical suspicion, and on an early initiation of immunosuppressive therapy. We did a nonsystematic review of the literature on autoimmune encephalitis between 2005 and 2017. We conclude that the prevalence of autoimmune encephalitis is increasing, even surpassing infectious causes of encephalitis in developed countries. Clinical presentation includes sub-acute cognitive and behavioral impairment, with or without alterations in consciousness and seizures. Fever and inflammation of the cerebrospinal fluid are less common than in the infectious causes but psychiatric symptoms are more frequent. There are specific clinical presentations according to the particular type of antigen/antibody present, which also determines the association with cancer, constituting a paraneoplastic syndrome only in some cases. Immunosuppressive therapy has been standardized in steps, and should be initiated early to improve prognosis.


The Symptom Checklist-Revised (SCL-90-R) is able to discriminate between simulation and fibromyalgia.
This study assessed the ability of the Symptom Checklist-Revised (SCL-90-R) to discriminate between two groups of fibromyalgia patients (those who were about to begin a treatment including the explicit aim of returning to work and those who were initiating a legal procedure to obtain permanent disability compensation) and two groups of healthy volunteers (medical students and psychology graduates), who were asked to produce a symptomatic resemblance to a chronic pain disorder. Logistic regression analyses were applied to the SCL-90-R subscales and individual probabilistic indices of simulation were calculated. Results showed that the SCL-90-R was able to discriminate between healthy subjects and both groups of patients with a high sensitivity and specificity. The individual indices of simulation, which might be more useful in clinical practice than the comparison of the SCL-90-R profiles, also showed an appropriate level of accuracy.


Place and contingency differential responses of monkey septal neurons during conditional place-object discrimination.
To elucidate spatial and cognitive function of the septal nuclei, neural activity was recorded from alert monkeys during performance of a place-dependent go/no-go (PGN) task. Response/reinforcement contingencies of given objects were conditional upon the location of a motorized, movable device (cab) containing a monkey in one of four places. The task was initiated by presentation of the outside view (place phase) followed by presentation of an object (object phase) selected from a total of four. A lever press was reinforced only if the correct object was seen in its corresponding place, and the same object was never reinforced in any of the other three places. Of 430 septal neurons recorded, the responses during the place phase in the four places were significantly different in 58 neurons. Responses of eight of these neurons were also place-differential during the object phase as well as the place phase. Furthermore, when the outside view was not presented before the object phase, differential responses in the object phase disappeared. Responses of 91 neurons in the object phase were differential in terms of go/no-go responses and reward availability. Of these 91 neurons, 72 were further tested on a place-independent asymmetrical go/no-go (AGN) task, which required no conditional discrimination. Forty-three neurons responded differentially only in the PGN task. It is thus concluded that this PGN-specific activity reflected conditional place-object relations. Of the remaining 29 neurons that responded differentially in both tasks, 21 were further tested by a place-independent symmetrical go/no-go task (no-go responses were also rewarded). Responses of 19 of these 21 neurons were related to the reward/nonreward contingency but not to the response contingency. The results suggest that septal nuclei are involved in integrating spatial information, conditional place-object relations, and reward/nonreward contingency.


Foot metastasis: Current knowledge.
Foot metastasis are rare and often overlooked due to non-specifical symptoms. This often leads to misdiagnosis delaying the right diagnosis. Metastatic disease of the foot is rare. Foot pain and swelling may be the presenting symptom of an occult malignancy. If metastatic disease is not kept in the differential diagnosis of foot pain, diagnosis and treatment will be delayed. The purpose of this study was to analyze articles presenting cases of foot metastasis to provide a more accurate incidence of symptomatic foot acrometastasis as well as to review the clinical course and outcomes. Studies were searched on PubMed/Medline from the inception to February 2020. All studies included in the review presented foot metastasis either with or without a known primary tumor. Most of the articles were case reports, to which we added two case reports of foot acrometastasis produced by our Institute. Forty-three studies with a total of 45 patients were included in this review. The literature published mostly concerning case reports about old patients (average age: 63,2), in a late phase of their disease. Lung cancer appeared to be the most common primary tumor, followed by endometrial and breast cancer. In the 36% of the cases foot metastasis were found when the primary site was still unknown. Calcaneus and metatarsal bones were the most common bones involved. Surgical solution is rare, the chosen treatments are often of palliative care. Prognosis was often poor, death occurred within 2 years since the discovery of foot metastasis in about 50% of cases.


Social Determinants of Depression: The Intersections of Race, Gender, and Socioeconomic Status.
Despite the wealth of literature on social determinants of mental health, less is known about the intersection of these determinants. Using a nationally representative sample, this study aimed to study separate, additive, and multiplicative effects of race, gender, and SES on the risk of major depressive episode (MDE) among American adults. National Survey of American Life (NSAL) included 3570 African Americans and 891 Whites. Race, gender, socioeconomic status (SES, household income, education, employment, and marital status) were independent variables. Twelve-month MDE was measured by the Composite International Diagnostic Interview (CIDI). A series of logistic regressions were used to analyze the data. In the pooled sample, race and household income, but not gender, education, employment, and marital status were associated with 12-month MDE. Gender interacted with the effects of income on MDE, suggesting that the association between household income and MDE is larger for women than men. In race by gender specific models that controlled for other SES indicators, high income was protective for White women, education was protective for African American women, and high income became a risk factor for African American men. High income did not show a risk effect for African American men in the absence of other SES indicators. Findings suggest that race, gender, and class interact on how SES indicators, such as education or income, become a protective or a risk factor for MDE among American Adults. When the outcome is MDE, White women benefit more from income, African American women gain from education, however, the residual effect of high income (above and beyond education, employment, and marital status) may become a risk factor for African American men.


Morphological observations on the replication of herpesvirus saimiri in monkey kidney cell cultures.
Owl and African green monkey kidney cell cultures have been infected with 1 p.f.u./cell of herpesvirus saimiri and sample cultures have been taken for examination by electron microscopy at 3 to 6 hourly intervals over a period of 7 days; the experiments were repeated several times. The peculiarly slow replication cycle of Herpesvirus saimiri has enabled distinct cytoplasmic and nuclear phases in virus maturation to be clearly distinguished; the overall fine structural features were similar in both cell types. Immature particles were first detected in the nucleus and cytoplasm 63 h after infection. Thereafter, abundant cytoplasmic immature particles matured by budding through cytoplasmic membranes until about 100 h, whereas nuclear immature particles budded through the inner nuclear membrane or intranuclear invaginations of it later, from about 100 h until cytolysis was complete at 160 h. Morphological differences were also observed between particles budding at cytoplasmic membranes and the nuclear envelope. At the former site the membrane overlying the bud showed an electron opaque thickening which imparted to the mature particle an asymmetrical appearance. Such thickenings of the envelope were not observed in mature particles of nuclear origin. Unusual tubular and laminated nuclear structures were seen towards the end of the replicative cycle corresponding with the phase of nuclear virus maturation by budding; the morphology of the latter structures is described.


beta-Lactam coselection of sensitive and TEM-1 beta-lactamase-producing subpopulations in heterogeneous Escherichia coli colonies.
We tested the effect of different concentrations of cefamandole, cefotaxime, cefoxitin and imipenem on homogeneous and heterogeneous bacterial stationary colonies. A reduction of not more than 100-fold in the bacterial colony population was obtained with sensitive Escherichia coli K12 strains. When an E. coli strain producing high levels of TEM-1 beta-lactamase constituted 6% or more of the population, the whole colony was resistant to cefamandole, but still sensitive to cefotaxime, cefoxitin or imipenem. Thus in the case of cefamandole the sensitive subpopulation is co-selected by the resistant one. It is suggested that the occurrence of resistant subpopulations, as a small part of an apparently sensitive E. coli population, can lead to therapeutic failures with beta-lactamase-sensitive agents.


Partition of alkane by an extracellular vesicle derived from hexadecane-grown Acinetobacter.
The enhanced solubility of hexadecane in the growth medium of hexadecane-grown Acinetobacter species has been related to the accumulation of an extracellular vesicular component. The partition of hexadecane was determined by measuring the amount of [3H]hexadecane bound to the vesicular particle. The vesicle was characterized as a phospholipid-rich, lipopolysaccharide-rich particle with a polypeptide composition similar to the outer membrane of Acinetobacter. The accumulation of an extracellular vesicular component that binds hexadecane in the form of a microemulsion represents another example of molecules produced by microorganisms in response to paraffinic substrates.


Molecular genetic and bile acid profiles in two Japanese patients with 3beta-hydroxy-DELTA5-C27-steroid dehydrogenase/isomerase deficiency.
We report definitive diagnosis and effective chenodeoxycholic acid (CDCA) treatment of two Japanese children with 3[beta]-hydroxy-[DELTA]5-C27-steroid dehydrogenase/isomerase deficiency. Findings of cholestasis with normal serum [gamma]-glutamyltransferase activity and total bile acid concentration indicated the need for definitive bile acid analysis. Large amounts of 3[beta]-hydroxy-[DELTA]5 bile acids were detected by gas chromatography-mass spectrometry. HSD3B7 gene analysis using peripheral lymphocyte genomic DNA from the patients and their parents identified four novel mutations of the HSD3B7 gene in the patients. One had a homozygous mutation, 314delA; the other had compound heterozygous mutations: V132F, T149I, and 973_974insCCTGC. Interestingly, the second patient's mother had V132F and T149I mutations in one allele. Excessive 3[beta]-hydroxy-[DELTA]5-bile acids such as 3[beta],7[alpha]-dihydroxy- and 3[beta],7[alpha],12[alpha]-trihydroxy-5-cholenoic acids were detected in the first patient's urine; the second patient's urine contained large amounts of 3[beta]-hydroxy-5-cholenoic acid. Liver dysfunction in both patients decreased with ursodeoxycholic acid treatment, but unusual bile acids were still detected. Normalization of the patients' liver function and improvement of bile acid profiles occurred with CDCA treatment. Thus, we found mutations in the HSD3B7 gene accounting for autosomal recessive neonatal cholestasis caused by 3[beta]-hydroxy-[DELTA]5-C27-steroid dehydrogenase/isomerase deficiency. Early neonatal diagnosis permits initiation of CDCA treatment at this critical time, before the late cholestatic stage.


A simple, quick one-step ELISA assay for the determination of complex plasma factor XIII (A2B2).
A new highly sensitive sandwich ELISA assay was developed for the determination of plasma factor XIII (FXIII). Plasma FXIII is a tetrameric complex of two types of subunits (A2B2). A biotinylated monoclonal capture-antibody against the B-subunit and a peroxidase-labelled monoclonal tag-antibody against the A-subunit were added to the plasma dilution and the amount of the complex attached to streptavidin-coated microplate was quantitated by measuring peroxidase activity. Only the tetrameric plasma FXIII reacted in the assay, non-complexed A or B subunits showed no reaction. The assay is linear up-to 40 microg/L of FXIII in the assay mixture. It is a quick one-step assay which can be performed within two hours. At normal and low FXIII concentration within batch reproducibility was 2.0% and 3.3%, day to day variation was 5.5% and 8.7%, respectively. Its high sensitivity allows reliable measurement at FXIII concentrations below 1% of normal average. Plasma samples can be stored for the assay at -20 degrees C for at least one month. Plasma levels of healthy individuals were normally distributed and no gender difference was observed. A reference interval of 14-28 mg/L (67-133%) was established.


Brief Report: Antibodies Reacting to Brain Tissue in Basque Spanish Children with Autism Spectrum Disorder and Their Mothers.
Previous investigations found that a subset of children with autism spectrum disorder (ASD) in California possessed plasma autoantibodies that reacted intensely with brain interneurons or other neural profiles. Moreover, for several cohorts of American women, maternal autoantibody reactivity to specific fetal brain proteins was highly specific to mothers of children with ASD. We sought to determine whether children and their mothers from a regionally specific cohort from the Basque Country of Spain demonstrated similar reactivity. Some children's plasma reacted to interneurons, beaded axons or other neural profiles with no difference in the occurrence of these antibodies in children with or without ASD. Findings on the maternal antibodies confirmed previous research; plasma reactivity to fetal brain a combination of proteins at 37 and 73 kDa or 39 and 73 kDa was found exclusively in mothers of children with ASD.


Sugar-Sweetened Beverage Consumption May Modify Associations Between Genetic Variants in the CHREBP (Carbohydrate Responsive Element Binding Protein) Locus and HDL-C (High-Density Lipoprotein Cholesterol) and Triglyceride Concentrations.
ChREBP (carbohydrate responsive element binding protein) is a transcription factor that responds to sugar consumption. Sugar-sweetened beverage (SSB) consumption and genetic variants in the CHREBP locus have separately been linked to HDL-C (high-density lipoprotein cholesterol) and triglyceride concentrations. We hypothesized that SSB consumption would modify the association between genetic variants in the CHREBP locus and dyslipidemia. Data from 11 cohorts from the Cohorts for Heart and Aging Research in Genomic Epidemiology consortium (N=63 599) and the UK Biobank (N=59 220) were used to quantify associations of SSB consumption, genetic variants, and their interaction on HDL-C and triglyceride concentrations using linear regression models. A total of 1606 single nucleotide polymorphisms within or near CHREBP were considered. SSB consumption was estimated from validated questionnaires, and participants were grouped by their estimated intake. In a meta-analysis, rs71556729 was significantly associated with higher HDL-C concentrations only among the highest SSB consumers (β, 2.12 [95% CI, 1.16-3.07] mg/dL per allele; P<0.0001), but not significantly among the lowest SSB consumers (P=0.81; PDiff <0.0001). Similar results were observed for 2 additional variants (rs35709627 and rs71556736). For triglyceride, rs55673514 was positively associated with triglyceride concentrations only among the highest SSB consumers (β, 0.06 [95% CI, 0.02-0.09] ln-mg/dL per allele, P=0.001) but not the lowest SSB consumers (P=0.84; PDiff=0.0005). Our results identified genetic variants in the CHREBP locus that may protect against SSB-associated reductions in HDL-C and other variants that may exacerbate SSB-associated increases in triglyceride concentrations. Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT00005133, NCT00005121, NCT00005487, and NCT00000479.


Reticular thalamic inhibitory input to lateral hypothalamic neurons: a functional and histochemical determination.
Both the lateral hypothalamus (LH) and reticular thalamus (RT) have been implicated in the integration of a variety of vital functions. To determine relevant connections more specifically between cells of these two regions in the rat, functional neurophysiological and horseradish peroxidase (HRP) histochemical techniques were utilized. Single pulse electrical stimulation of the RT modified the discharge frequency of the majority of LH neurons tested. LH neuronal responses to RT stimulation included; inhibition (66.32%), excitation followed by inhibition (6.12%), and excitation (7.14%). Because RT stimulation resulted primarily in the inhibition of LH neuronal activity with a relative short mean latency of 1.87 +/- 0.23 ms, the existence of a possible direct inhibitory pathway from the RT to LH also was evaluated using HRP histochemistry. Retrogradely HPR labeled soma were identified in the RT after HRP was ejected extracellularly onto LH neurons, which exhibited a decrease in spontaneous activity in response to RT stimulation. These data demonstrate direct projections from the RT to the LH. In addition, HRP labeling of neurons and axons in the zona incerta-LH area following LH HRP ejections suggest this route as a synaptic pathway from RT to LH.


Cost analyses of obesity in Canada: scope, quality, and implications.
Rapid changes in lifestyle have led to a global obesity epidemic. Understanding the economic burden associated with the obesity epidemic is essential to decision making of cost-effective interventions. This study reviewed costs of obesity and intervention programs in Canada, assessed the scope and quality of existing cost analyses, and identified implications for economic evaluations and public health decision makers. A systematic search of costs associated with obesity or intervention program in Canada between 1990 and 2011 yielded 10 English language articles eligible for review. The majority of studies was prevalence-based or top-down costing; 40% had excellent quality assessed using the Quality of Health Economic Study scale. The aggregated annual costs of obesity in Canada ranged from 1.27 to 11.08 billion dollars. Direct costs accounted for 37.2% to 54.5% of total annual costs. Between 2.2% and 12.0% of Canada's total health expenditures were attributable to obesity. The average annual physician cost of overweight male ($ 427) and female ($ 578) adults was lower than that of obese male ($ 475) and female ($ 682) adults; this cost differential across weight status groups was comparable to that found in adolescents. The cost for implementation and maintenance of a school-based obesity prevention program was $ 23 per student. We observed high costs associated with overweight and obesity and modest costs for obesity prevention programs; however, no cost-effectiveness study of obesity interventions has been performed in Canada. Cost-effectiveness analyses of preventive programs that constitute incidence-based life-time modeling of costs and health outcomes from societal perspective are urgently needed.


Bias in the measurement of the outcome is associated with effect sizes in randomized clinical trials on exercise therapy for chronic low back pain: a meta-epidemiological study.
To explore the relationships between the risk of bias and treatment effect estimates for exercise therapy interventions on pain intensity and physical functioning outcomes in randomized controlled trials (RCTs) involving patients with chronic low back pain. A cross-sectional meta-epidemiological study of the 230 RCTs (31,674 participants) in the 2021 'Exercise therapy for chronic low back pain' Cochrane Review were included. Study design characteristics, sample size, prospective trial registration, flowchart information, interventions, and comparisons were extracted. Independent pairs of reviewers assessed the risk of bias using the Cochrane Risk of Bias 2 tool. The metaregression included 220 (pain intensity) and 203 (physical functioning) effect sizes. Unadjusted and adjusted metaregression models showed no significant associations between the bias domains and pain intensity effect sizes. Only domain 'bias in the measurement of the outcome' was significantly associated with physical functioning (standardized mean difference: -0.40, 95% confidence interval: -0.77 to -0.02) when adjusted for flowchart reported (yes/no), prospective trial registration, sample size, and comparator type. The risk of bias in the measurement of the outcome could lead to slight overestimates of the effect size for physical functioning. Clinicians should consider this when they read and assess RCT results in this field. We encourage metaresearchers to replicate our findings using a consistent approach for evaluating the risk of bias (i.e., the RoB 2 tool) in other musculoskeletal conditions and interventions to investigate their generalizability.


ADCC employing an NK cell line (haNK) expressing the high affinity CD16 allele with avelumab, an anti-PD-L1 antibody.
NK-92 cells, and their derivative, designated aNK, were obtained from a patient with non-Hodgkin lymphoma. Prior clinical studies employing adoptively transferred irradiated aNK cells have provided evidence of clinical benefit and an acceptable safety profile. aNK cells have now been engineered to express IL-2 and the high affinity (ha) CD16 allele (designated haNK). Avelumab is a human IgG1 anti-PD-L1 monoclonal antibody, which has shown evidence of clinical activity in a range of human tumors. Prior in vitro studies have shown that avelumab has the ability to mediate antibody-dependent cell-mediated cytotoxicity (ADCC) of human tumor cells when combined with NK cells. In the studies reported here, the ability of avelumab to enhance the lysis of a range of human carcinoma cells by irradiated haNK cells via the ADCC mechanism is demonstrated; this ADCC is shown to be inhibited by anti-CD16 blocking antibody and by concanamycin A, indicating the use of the granzyme/perforin pathway in tumor cell lysis. Studies also show that while NK cells have the ability to lyse aNK or haNK cells, the addition of NK cells to irradiated haNK cells does not inhibit haNK-mediated lysis of human tumor cells, with or without the addition of avelumab. Avelumab-mediated lysis of tumor cells by irradiated haNK cells is also shown to be similar to that of NK cells bearing the V/V Fc receptor high affinity allele. These studies thus provide the rationale for the clinical evaluation of the combined use of avelumab with that of irradiated adoptively transferred haNK cells.


Recurrent BRCA2 exon 3 deletion in Assyrian families.
We identified six patients from five families with a recurrent mutation: NM_000059.3 (BRCA2) exon 3 deletion. All families self-identified as Assyrian. Assyrians are an ethnoreligious population of ancient Mesopotamia, now mostly living in modern day Iraq, Syria, Turkey and Iran. They are historically a socially isolated population with intermarriage within their community, living as a religious and language minority in mostly Muslim countries. The probands of each family presented with a classic BRCA2-associated cancer including early-onset breast cancer, epithelial serous ovarian cancer, male breast cancer and/or high-grade prostate cancer, and family history that was also significant for BRCA2-associated cancer. BRCA2 exon 3 deletion is classified as pathogenic and has been previously described in the literature, but it has not been described as a founder mutation in a particular population. We characterise this recurrent BRCA2 pathogenic variant in five Assyrian families in a single centre cohort.


Mechanochromic Stretchable Electronics.
Soft and stretchable electronics are promising for a variety of applications such as wearable electronics, human-machine interfaces, and soft robotics. These devices, which are often encased in elastomeric materials, maintain or adjust their functionality during deformation, but can fail catastrophically if extended too far. Here, we report new functional composites in which stretchable electronic properties are coupled to molecular mechanochromic function, enabling at-a-glance visual cues that inform user control. These properties are realized by covalently incorporating a spiropyran mechanophore within poly(dimethylsiloxane) to indicate with a visible color change that a strain threshold has been reached. The resulting colorimetric elastomers can be molded and patterned so that, for example, the word "STOP" appears when a critical strain is reached, indicating to the user that further strain risks device failure. We also show that the strain at color onset can be controlled by layering silicones with different moduli into a composite. As a demonstration, we show how color onset can be tailored to indicate a when a specified frequency of a stretchable liquid metal antenna has been reached. The multiscale combination of mechanochromism and soft electronics offers a new avenue to empower user control of strain-dependent properties for future stretchable devices.


Benefits and effectiveness of using a wrist heart rate monitor as a telerehabilitation device in cardiac patients: A randomized controlled trial.
Telerehabilitation in cardiology has the potential to become the alternative to regular outpatient cardiac rehabilitation. Our study focuses on the wrist heart rate monitor as a telerehabilitation device, defines detected limitations, and compares results between home-based and regular outpatient rehabilitation methods, related to physical fitness, quality of life, and training adherence. The study design was a randomized controlled trial. Eligible 56 cardiac rehabilitation patients were randomized into a 12-week regular outpatient training group (ROT) and interventional home-based telerehabilitation group (ITG). For both groups, the intensity of the training was prescribed to be performed at 70% to 80% of heart rate reserve for 60 minutes, 3 times a week. The ITG patients started their training with a wrist heart rate monitor in their home environment. These patients received feedback once a week, reflecting data uploaded on the internet application. The ROT patients performed their exercise under the direct supervision of a physical specialist in a regular outpatient clinic. Physical fitness and health-related quality of life were assessed at baseline and after 12 weeks. Training adherence in both groups was determined and compared. Fifty-one patients comleted the intervention (91%); no serious adverse events were recorded. Physical fitness expressed as peak oxygen uptake showed significant improvement (P < .001) in ROT group from 23.4 ± 3.3 to 25.9 ± 4.1 mL/kg/min and (P < .01) in ITG group from 23.7 ± 4.1 to 26.5 ± 5.7 mL/kg/min without significant between-group differences after 12 weeks of intervention. The training adherence between groups was similar. Our study shows that telerehabilitation via wrist heart rate monitor could become an alternative kind of cardiac rehabilitation which deserves attention and further analyzing.


Sex education and rehabilitation with schizophrenic male outpatients.
Research indicates that schizophrenic patients lack intimate relationships and show a high rate of sexual dysfunction. Despite increasing awareness of the rights of handicapped persons to sexual expression, the treatment of schizophrenic patients rarely addresses their sexuality. A sex education program for recent-onset male schizophrenic patients attending an outpatient clinic was developed in response to several incidents involving patients' inappropriate sexual behaviors. To enhance our understanding of the current sexual functioning and needs of these patients, sex histories were taken. Almost all of the 16 patients interviewed were sexually active, with autoerotic activity predominating. Sixty-three percent of the patients reported orgasmic and/or erectile dysfunctions. Other studies have linked sexual dysfunction to the side effects of antipsychotic medications. The objectives of the sex education program were: to provide information; to clarify values; to overcome sexual dysfunction; and to enhance intimacy skills. The authors used role playing, modeling, group exercises, and explicit sex therapy audiovisual material to improve patients' intimacy skills. Patients participated actively and used the group to explore sexual issues. No exacerbations of symptoms were observed among patients participating in the program.