Single-cell transcriptional diversity of neonatal umbilical cord blood immune cells reveals neonatal immune tolerance.
The characteristics of neonatal immune cells display intrinsic differences compared with adult immune cells. Therefore, a comprehensive analysis of key gene expression regulation is required to understand the response of the human fetal immune system to infections. Here, we applied single-cell RNA sequencing (scRNA-seq) and single-cell sequencing assay for transposase-accessible chromatin (scATAC-seq) to systematically profile umbilical cord blood (UCB) nucleated cells and peripheral blood mononuclear cells (PBMCs) to identify their composition and differentially expressed genes. The immune cells in neonatal UCB demonstrated the expression of key genes, such as HBG2, NFKBIA, JUN, FOS, and TNFAIP3. In contrast, natural killer and T cells, which are constituents of adult PBMCs, exhibited high cytotoxic gene expression. Furthermore, we obtained similar results from the data of scATAC-seq by identifying the status of chromatin accessibility of key genes. Therefore, scRNA-seq and scATAC-seq of neonatal UCB nucleated cells and adult PBMCs could serve as an invaluable resource for elucidating the regulatory mechanisms of responses of distinct immune cell types and further identifying the differences between neonatal and adult immune responses to predict the potential underlying mechanism for neonatal immune tolerance.


An investigation of the relationship between emotional maladjustment and infertility.
The purpose of this investigation was to examine the psychologic test results of women diagnosed with infertility as compared with the general population of women. The infertile group consisted of 150 women, whereas the control group included 50 women. Both groups were similar in regard to age and number of years married. The tests administered were: the 16 Personality Factor, the IPAT Anxiety Scale, the IPAT Depression Scale, the Tennessee Self-Concept Scale, and the Internal-External Scale (locus of control). Scores on a total of 41 test variables did not differ significantly between the two groups. In fact, the means and standard deviations for all variables were remarkably similar. This study concludes that significant emotional maladjustment is no more prevalent in women coping with infertility than for the general population of women. Results from this investigation cast doubt on the historical assumption that stress may be a causal factor in infertility.


Acute effects of 4-ipomeanol on experimental lung tumors with bronchiolar or alveolar cell features in Syrian hamsters or C3H/HeNCr mice.
4-Ipomenaol (IPO) has been shown to induce P-450-mediated necrosis of Clara cells in experimental animals, and clinical trials were initiated to treat people with bronchioloalveolar cancers with this novel drug. We therefore performed experiments to examine two different animal lung tumor models for acute IPO cytotoxicity: hamster Clara-cell-derived adenocarcinomas and mouse alveolar type II cell tumors. Clara cells serve as stem cells for airway cell renewal and, therefore, tumors derived from Clara cells may likewise differentiate into various bronchiolar cell types, or undergo squamous cell metaplasia. Bronchiolar cell tumors were induced in Syrian hamsters by a single weekly gavage with 6.8 mg N-nitrosomethyl-n-heptylamine (NMHA)/animal for 35 weeks. NMHA-induced bronchiolar tumors were classified as well-differentiated lepidic bronchioloalveolar carcinomas, acinar adenocarcinoma, adenosquamous carcinoma, and squamous-cell carcinoma. After 35 and 46 experimental weeks, control and carcinogen-treated hamsters were injected once with doses of 40-110 mg IPO/kg i.p. and necropsied 15-48 h later. Solid and papillary tumors with alveolar cell features were induced transplacentally in C3H/HeNCr mice, by treating pregnant animals on gestation day 16 with 0.5 mmol N-nitrosoethylurea/kg, i.p. Offspring of control and carcinogen-treated mice were injected at 2-3 months of age with 35 mg or 50 mg IPO/kg i.p. and necropsied either 24-48 h or 5 and 12 days after injection. Light microscopic studies were carried out to assess cytotoxic effects in various tissues in both hamsters and mice; in hamsters, additional ultrastructural studies were performed. When administered to hamsters, IPO induced moderate to severe cytotoxicity in normal and dysplastic bronchiolar lining cells, in most lepidic bronchioloalveolar carcinomas, and in some glandular areas of adenosquamous cell carcinomas. Susceptible cells included normal, anaplastic, and neoplastic nonciliated and some ciliated bronchiolar cells. Undifferentiated and squamous tumor cells were resistant to IPO, as were resident normal alveolar type II cells. However, some adenocarcinomas composed primarily of ciliated and mucous cells also showed no IPO-induced necrosis, indicating a deficiency in appropriate activating enzymes. In the mice, IPO induced bronchiolar cell necrosis and, at the high dose, also severe pulmonary edema. No cytotoxicity was observed in normal or hyperplastic alveolar epithelium, nor in either solid or papillary growth forms of mouse alveolar cell tumors.(ABSTRACT TRUNCATED AT 400 WORDS)


Patient and caregiver opinions about an FDA-developed generic drug educational handout: A pilot cross-sectional survey.
To examine patient and caregiver opinions and "receptivity" regarding generic drug educational material in terms of content, format and design, delivery channel, and level of satisfaction. Interviewer-administered surveys were conducted with patients and caregivers who were clients of a regional medication management program or pharmacy services clinic to gather perceptions about generic drugs and input on a generic drug educational handout developed by the U.S. Food and Drug Administration (FDA). Survey questions were developed by the investigators and pretested before use. Data were analyzed using descriptive statistics, and responses to open-ended questions were assessed using qualitative content analysis. Survey psychometrics were examined using exploratory factor analysis (EFA). Of the 100 survey participants, most (93%) had positive perceptions about generic drug safety and effectiveness after reading the handout. Most participants were satisfied or very satisfied with the handout's content (87%) as well as format and design (92%). However, more than 20% of participants were still unsure about the benefits and risks of generic drugs compared with those of brand drugs, and more than 15% were still unsure about which benefits and risks mattered most to them. The participants' preferred source for the handout was a pharmacy (49%) or doctor's office (27%). In an EFA of the survey instrument, 2 factors emerged related to the educational handout's content: (1) generic drug information, a 7-item factor (Cronbach alpha = 0.882); and (2) personal relevance, a 3-item factor (Cronbach alpha = 0.692). The findings indicate an overall positive "receptiveness" toward generic drug informational materials from patients and caregivers, which highlights the feasibility and importance of educational outreach programs about generic drugs targeted toward this population. Future studies may focus on more diverse populations and tailor materials to the needs of specific patient and caregiver subgroups and health literacy levels.


Second stem cell transplantation in patients with Fanconi anemia using antithymocyte globulin alone for conditioning.
Despite the promising data on the outcome of allogeneic stem cell transplantation (SCT) in patients with Fanconi anemia (FA), a certain percentage of these patients still experience graft failure; some of these patients undergo second transplants, but the existing data on the outcome of the second SCT in FA patients are scarce, with no long-term follow-up provided in many of the publications addressing this issue. This is a review of our experience in 4 such patients who underwent second stem cell transplants using rabbit ATG only for conditioning. Three engrafted promptly and are alive and free of disease at 25, 23, and 21 months, respectively. We conclude, therefore, that the use of ATG alone for conditioning before a second SCT may offer a chance of long-term disease-free survival for FA patients who fail their first transplant.


Optimal Positioning for Volar Plate Fixation of a Distal Radius Fracture: Determining the Distal Dorsal Cortical Distance.
Distal radius fractures are currently among the most common fractures of the musculoskeletal system. With a population that is living longer, being more active, and the increasing incidence of osteoporosis, these injuries will continue to become increasingly prevalent. When operative fixation is indicated, the volar locking plate has recently become the treatment of choice. However, despite its success, suboptimal position of the volar locking plate can still result in radiographic loss of reduction. The distal dorsal cortical distance is being introduced as an intraoperative radiographic tool to help optimize plate position and minimize late loss of fracture reduction.


Propranolol for the Treatment of Hemangioma in PHACE Syndrome: A Case Report.
Oral propranolol is commonly used as a first-line treatment for infantile hemangioma. However, its use in PHACE (posterior fossa anomalies, hemangioma, arterial anomalies, cardiac anomalies, and eye anomalies) syndrome raises concerns that it might exacerbate the patient's risk of stroke. Here, we report the case of a four-month-old premature girl with PHACE syndrome, who presented with a large hemangioma involving the left side of her face, following the V1+V2+V3 distribution, including the upper lip, left ear, and left eye. This condition was successfully treated with propranolol, and no adverse side effects were reported.


Measurement of skeletal muscle perfusion during postischemic reactive hyperemia using contrast-enhanced MRI with a step-input function.
The regional distribution of skeletal muscle blood flow was measured during postischemic reactive hyperemia using Gd-DTPA contrast-enhanced (CE) MRI. The release of an occlusive thigh cuff was used to deliver a step-input of contrast concentration that was coincident with the onset of reactive hyperemia. A first-order tracer kinetic equation was used to estimate the unidirectional influx constant, Ki (ml/100 g/min), and the distribution volume of Gd-DTPA in the tissue, v(e), from T1-weighted images acquired with saturation recovery (SR) steady-state free precession (SSFP) and spoiled gradient-echo (SPGR) protocols. The capillary permeability surface (PS) area increased significantly during reactive hyperemia, which facilitated rapid extraction of Gd-DTPA during the first pass. Regional muscle group studies from 11 normal volunteers yielded blood flow (Ki) values of 108.3 +/- 34.1 ml/100 g/min in the gastrocnemius, 184.3 +/- 41.3 ml/100 g/min in the soleus, and 122.4 +/- 34.4 ml/100 g/min in the tibialis anterior. The distribution volumes (v(e)) in the corresponding muscle groups were respectively 8.3% +/- 2.1%, 9.3% +/- 1.9%, and 7.9% +/- 1.8% from the kinetic model, and 8.8% +/- 2.4%, 9.1% +/- 1.9%, and 7.2% +/- 1.4% from tissue relaxometry studies. Bulk blood flow studies in the same volunteers using phase-contrast velocimetry (popliteal artery) yielded significantly lower flow values, but with a correlation coefficient R2 = 0.62 and P = 0.004.


Retraction: Li, X.; et al. Circular RNA circ-FoxO3 Inhibits Myoblast Cells Differentiation. Cells 2019, 8, 616.
It has come to our attention that two images in Figure 2A (100%), and Figure 2C (GM) of the manuscript [...].


The reform of UK research ethics committees: throwing the baby out with the bath water?
On 1 May 2004 research ethics committees became legally accountable to a new government body, the United Kingdom Ethics Committee Authority. This marks the end of the self regulation of research ethics. This paper describes how this change in research ethics committee status has come about and explores the implications for research subjects, researchers, institutions, and for regulation of research.


Induction of intracellular and extracellular beta-galactosidase activity in Phycomyces blakesleeanus.
The inductive effect of different sugars on beta-galactosidase synthesis in Phycomyces blakesleeanus has been studied. The enzyme was inducible by galactose and fructose. When grown on these sugars the enzyme level was 10-20 times greater than when grown on glucose. We have detected both intra- and extracellular beta-galactosidase activity when Phycomyces blakesleeanus was grown on galactose, but only extracellular beta-galactosidase activity when grown on fructose plus lactose.


Systematic transcriptomic and phenotypic characterization of human and murine cardiac myocyte cell lines and primary cardiomyocytes reveals serious limitations and low resemblances to adult cardiac phenotype.
Cardiac cell lines and primary cells are widely used in cardiovascular research. Despite increasing number of publications using these models, comparative characterization of these cell lines has not been performed, therefore, their limitations are undetermined. We aimed to compare cardiac cell lines to primary cardiomyocytes and to mature cardiac tissues in a systematic manner. Cardiac cell lines (H9C2, AC16, HL-1) were differentiated with widely used protocols. Left ventricular tissue, neonatal primary cardiomyocytes, and human induced pluripotent stem cell-derived cardiomyocytes served as reference tissue or cells. RNA expression of cardiac markers (e.g. Tnnt2, Ryr2) was markedly lower in cell lines compared to references. Differentiation induced increase in cardiac- and decrease in embryonic markers however, the overall transcriptomic profile and annotation to relevant biological processes showed consistently less pronounced cardiac phenotype in all cell lines in comparison to the corresponding references. Immunocytochemistry confirmed low expressions of structural protein sarcomeric alpha-actinin, troponin I and caveolin-3 in cell lines. Susceptibility of cell lines to sI/R injury in terms of viability as well as mitochondrial polarization differed from the primary cells irrespective of their degree of differentiation. Expression patterns of cardiomyocyte markers and whole transcriptomic profile, as well as response to sI/R, and to hypertrophic stimuli indicate low-to-moderate similarity of cell lines to primary cells/cardiac tissues regardless their differentiation. Low resemblance of cell lines to mature adult cardiac tissue limits their potential use. Low translational value should be taken into account while choosing a particular cell line to model cardiomyocytes.


Multimodal confocal microscopy for diagnosing nonmelanoma skin cancers.
The standard diagnostic procedure for skin cancers is invasive biopsy followed by histopathological evaluation. The biopsy may result in scarring and infection. A reliable way to noninvasively image suspicious lesions with high resolution and contrast would be valuable. In this study, the suitability of dye-enhanced multimodal confocal microscopy for the detection of nonmelanoma skin cancers was evaluated. For the experiments we used fresh tumor material stained using 0.2 mg/ml or 0.05 mg/ml aqueous solutions of methylene blue (MB) or toluidine blue (TB), respectively. Reflectance, fluorescence, and fluorescence polarization images of skin specimens stained with MB and TB were excited by 656 nm and 633 nm light, respectively. Fluorescence emission and anisotropy were registered between 690 nm and 710 nm. In addition, reference reflectance images at 830 nm were acquired. In total we imaged, analyzed, and compared to histology at least 10 samples of each tumor-type including nodular basal cell carcinoma (BCC), infiltrative basal cell carcinoma, and squamous cell carcinoma (SCC). The morphological features and appearance of skin structures in the fluorescence images correlate well with corresponding histology for all investigated tumor-types. Multi-spectral reflectance images provide information on the tissue spectral responses and are complimentary to the fluorescence images. The differences detected by fluorescence polarization in cancerous and normal structures may be used for cancerous tissue discrimination. Our results indicate the feasibility of using multimodal confocal microscopy as real-time tool for detecting skin pathology.


Evolution of tumor cytogenetic aberrations and N-myc oncogene amplification in a case of disseminated neuroblastoma.
Chromosome analyses including in situ hybridization with the protooncogene N-myc were carried out at diagnosis and several times at relapse in the bone marrow cells of a girl with disseminated neuroblastoma (stage IV) at ages 9, 23, 24, and 26 months, respectively. The tumor karyotype was pseudodiploid exhibiting partial monosomy for the short arm of chromosome #1, an aberration of the short arm of chromosome #2, and double minutes at diagnosis. Further structural aberrations of chromosomes #4, #6, and #17, and homogeneously staining regions could be demonstrated at relapse. In particular, the following structural aberrations were encountered: t(1;?)(p22;?),t(2;?)(p24;?), t(4;6)(q31;q25), and a der(17q). All tumor metaphases without homogeneously staining regions contained double minutes. N-myc, normally positioned within bands 2p23-24, was found to be amplified in a homogeneously staining region on the short arm of chromosome #15 by in situ hybridization. It is speculated that the translocation t(2;?)(p24;?) might be related to N-myc oncogene activation and subsequent amplification.


Heuweltjies (earth mounds) in the Clanwilliam district, Cape Province, South Africa: 4000-year-old termite nests.
Examination of eroded and intact earth mounds in the Clanwilliam district, South Africa, indicates that they are well-established active termitaria of the harvester termite Microhodotermes viator. Unoccupied lower portions of the mounds contain ubiquitous trace-fossil evidence of earlier inhabitation by the same species. Previous studies indicating that fossorial molerats played a major role in the formation of the mounds are not supported by the observations presented here. Calcretization of the basal parts of the earth mounds has been caused by groundwater interaction with the more alkaline mound soil. 14C dating of this calcrete indicates that the earth mounds have been in existence for at least 4000 years, an order of magnitude greater than any previously recorded longevity for termitarium inhabitation.


Enhancer-driven transcription of MCM8 by E2F4 promotes ATR pathway activation and glioma stem cell characteristics.
Glioma stem cells (GSCs) are responsible for glioma recurrence and drug resistance, yet the mechanisms underlying their maintenance remains unclear. This study aimed to identify enhancer-controlled genes involved in GSCs maintenance and elucidate the mechanisms underlying their regulation. We analyzed RNA-seq data and H3K27ac ChIP-seq data from GSE119776 to identify differentially expressed genes and enhancers, respectively. Gene Ontology analysis was performed for functional enrichment. Transcription factors were predicted using the Toolkit for Cistrome Data Browser. Prognostic analysis and gene expression correlation was conducted using the Chinese Glioma Genome Atlas (CGGA) data. Two GSC cell lines, GSC-A172 and GSC-U138MG, were isolated from A172 and U138MG cell lines. qRT-PCR was used to detect gene transcription levels. ChIP-qPCR was used to detect H3K27ac of enhancers, and binding of E2F4 to target gene enhancers. Western blot was used to analyze protein levels of p-ATR and γH2AX. Sphere formation, limiting dilution and cell growth assays were used to analyze GSCs growth and self-renewal. We found that upregulated genes in GSCs were associated with ataxia-telangiectasia-mutated-and-Rad3-related kinase (ATR) pathway activation, and that seven enhancer-controlled genes related to ATR pathway activation (LIN9, MCM8, CEP72, POLA1, DBF4, NDE1, and CDKN2C) were identified. Expression of these genes corresponded to poor prognosis in glioma patients. E2F4 was identified as a transcription factor that regulates enhancer-controlled genes related to the ATR pathway activation, with MCM8 having the highest hazard ratio among genes positively correlated with E2F4 expression. E2F4 bound to MCM8 enhancers to promote its transcription. Overexpression of MCM8 partially restored the inhibition of GSCs self-renewal, cell growth, and the ATR pathway activation caused by E2F4 knockdown. Our study demonstrated that E2F4-mediated enhancer activation of MCM8 promotes the ATR pathway activation and GSCs characteristics. These findings offer promising targets for the development of new therapies for gliomas.


Preparation and evaluation of Bletilla striata polysaccharide/graphene oxide composite hemostatic sponge.
Uncontrolled bleeding is an important cause of military and civilian casualties. GO has received more attention in the field of hemostasis. However, pure GO has various limitation in application due to its potential thrombosis, hemolytic and cytotoxicity. Herein, we present a simple, rapid and low-cost method to combine GO and natural polysaccharides by hydrogen bonding to prepare a new material Bletilla striata polysaccharide/graphene oxide composite sponge (BGCS). The BGCS was successfully synthesized and characterized by SEM, IR, RAMAN, XRD and Zeta potential analyzer analysis. The BGCS exhibited favorable biocompatibility. Besides, the porosity of BGCS was higher than 90% and showed good water absorption capacity. The results of whole blood coagulation evaluation showed that the BGCS can promote blood coagulation within 30 s without anticoagulant, showing excellent hemostatic effect. Further coagulation mechanism studies indicated that the surface of the BGCS possessed a high charge (-27.3 ± 0.9 mV) and showed strong platelet stimulation, the BGCS can also induce red blood cell aggregation, accelerate fibrin formation and accelerate blood coagulation. Therefore, the BGCS can stop bleeding within 50 s in rat-tail amputation models. The BGCS provides a new perspective for the safe application of GO in the field of trauma hemostasis.


[Osteoimmunology - IMMUNOBONE - Understanding the interaction between bone and immune system].
The special program "Osteoimmunology" under the leadership of the Universities of Erlangen, Jena, Gießen and Münster, is investigating in 27 projects clarification approaches on the causes of inflammatory rheumatic diseases, such as rheumatoid arthritis and ankylosing spondylitis with the aim of developing new forms of treatment. The molecular mechanisms involved in bone damage and the interaction between the immune system and bone and cartilage are topics of research.


Autoantibody-Mediated Macrophage Responses Provide the Missing Link between Innate and Adaptive Immune Dysfunction in Hidradenitis Suppurativa.
Hidradenitis suppurativa is considered to be a T helper 17-mediated inflammatory disorder. However, the role of prominent B-cell and plasma cell infiltrates has not been incorporated into pathogenic understanding of the disease. In their new article in the Journal of Investigative Dermatology, Carmona-Rivera et al. (2021) present new insights regarding autoantibody-mediated macrophage activation, which bridges the link between the innate and adaptive immune responses in severe hidradenitis suppurativa.


Morula-like cells in photo-symbiotic clams harboring zooxanthellae.
Symbiosis is observed between zooxanthellae, symbiotic dinoflagellates, and giant clams and related clams which belong to the families Tridacnidae and Cardiidae. We have previously shown that a photo-symbiotic clam Tridacna crocea has three types of hemocytes, the eosinophilic granular hemocyte with phagocytic activity, the agranular cell with electron lucent granules, and the morula-like cell with large (ca. 2 mum in diameter) colorless granules. The function of the morula-like cell is not clear, but it has not been reported in any other bivalves except photo-symbiotic clams T. crocea and Tridacna maxima. In order to clarify whether it is specific to photo-symbiotic clams or not, we studied hemocytes in the photo-symbiotic clams Tridacna derasa (Tridacnidae), Hippopus hippopus (Tridacnidae) and Corculum cardissa (Cardiidae), and a closely related non-symbiotic clam Fulvia mutica (Cardiidae). The eosinophilic granular hemocytes and the agranular cells were found in all of the clams examined. However, the morula-like cells which were packed with many large electron dense granules (ca. 2 mum in diameter), were observed only in the photo-symbiotic clams. In F. mutica, a closely related non-symbiotic clam, this type of hemocyte was not found. Instead a hemocyte with vacuoles and a few large granules containing peroxidase activity was observed. The large granules of F. mutica varied in size from ca. 1-9 mum in diameter. Present data suggests that the presence of morula-like cells is restricted to photo-symbiotic clams and that the hemocytes associated with the morula-like cells may have some functional relationship to symbiosis with zooxanthellae.


Bilateral areolar hyperpigmentation following transdermal application of compounded hormone replacement therapy.
A 61 year-old post-menopausal Caucasian woman presented to the pharmacy to re-initiate hormone therapy after having discontinued treatment for three years. She had discontinued previous hormone therapy due to severe hyperpigmentation of both nipples and areola. She stated that her nipples and areola had turned black, as if they had been burned. This reaction seemed unique, and we felt that it warranted further investigation.


The combined effects of unilateral enucleation and rearing in a "dim" red light on synapse-to-neuron ratios in the rat superior colliculus.
Rats whose right eyes were enucleated on day 1 after birth and nonenucleated rats were raised in either "light" or "dark" (red light) conditions from birth until 39 days of age. This resulted in 4 groups of animals: light-reared nonenucleated, light-reared enucleated, dark-reared nonenucleated, and dark-reared enucleated. At 39 days of age, the animals were killed by perfusion with 2.5% sodium cacodylate buffered glutaraldehyde. The superior colliculi were dissected out and processed for embedding in resin. Stereological procedures at the light and electron microscopical levels were used to estimate the synapse-to-neuron ratios in the superficial layers of these colliculi. Light-reared, nonenucleated rats had about 1,850 synapses-per-neuron in both the right and left superior colliculi. Rearing nonenucleated rats in the dark reduced this value to about 1,200. Enucleated rats reared in the light showed a differential response in the 2 colliculi. Thus, the contralateral (to the enucleated eye) colliculi showed a decrease, whereas the ipsilateral colliculi showed an increase in the synapse-to-neuron ratio compared with light-reared, nonenucleated rats. When enucleated rats were reared in the red light, there was a decrease in the ratio in both colliculi, although the extent of this decrease was more marked in the contralateral than the ipsilateral colliculi. However, the decrease in the contralateral colliculi was not significantly greater than that observed in the corresponding colliculi from dark-reared, nonenucleated rats. These results provide useful information on the combined and separate effects of unilateral enucleation at around birth and dark (red light) rearing during early life on the interneuronal connectivity of both the ipsi- and contralateral superior colliculi of rats. They also show the vast importance of visual stimulation for the normal development of the subcortical visual centers.


Determination of phytate in high molecular weight, charged organic matrices by two-dimensional size exclusion-ion chromatography.
A two-dimensional chromatography method for analyzing phytate or other ionic targets in matrices containing high molecular weight, charged organic species is described. Prior to quantification by anion exchange chromatography, the sample matrix was prepared by size exclusion chromatography, which removed the majority of the matrix. Quantification of phytate on the AS11-HC was sensitive (0.25µM, 0.17mg/l) and reproducible (4.6% RSD) allowing this method to provide baseline separation of phytate from a manure extract within 14min. The method is simple, requiring only sample filtering, reproducible (between-run variation 5% RSD) and linear from 0.38 to 76µM (0.25-50mg/l). The method is suitable for routine determination of phytate in high organic matrices such as manure extracts.


Indolent systemic mastocytosis associated with atypical small lymphocytic lymphoma: a rare form of concomitant lymphoproliferative disease.
Patients with systemic mastocytosis (SM) may acquire an associated hematologic non-mast cell (MC)-lineage disease (AHNMD). In most cases, a myeloid neoplasm is diagnosed, whereas the occurrence of a lymphoproliferative disease is an extremely rare event. We report on a patient with indolent SM associated with small lymphocytic lymphoma (SLL). The patient presented with lymphadenopathy, maculopapular exanthema, and elevated serum tryptase. The bone marrow biopsy showed focal MC aggregates together with SLL. As assessed by immunostaining, neoplastic MC were found to exhibit CD117 and CD25 but did not display CD5 or CD20, whereas SLL cells were found to coexpress CD5 and CD20 but did not express MC antigens. The KIT mutation D816V was detected in sorted CD34(+) cells and unfractionated marrow cells but not in CD5(+) SLL cells, confirming the coexistence of 2 distinct neoplasms.


Corrigendum: Patients with myeloproliferative neoplasms harbor high frequencies of CD8 T cell-platelet aggregates associated with T cell suppression.
[This corrects the article DOI: 10.3389/fimmu.2022.866610.].


ESR1 polymorphism is associated with plasma lipid and apolipoprotein levels in Caucasians of the Rochester Family Heart Study.
We evaluated six estrogen receptor 1 (ESR1) polymorphisms for association with ten plasma lipid and apolipoprotein traits in 1,847 individuals (941 females and 906 males) in the multi-generation Rochester Family Heart Study using a generalized estimating equation approach. Apolipoprotein A-I (apoA-I), apoA-II, and HDL-cholesterol (HDL-C) were associated with exon 4 rs1801132 (Pro325Pro) genotype (P = 0.0044, P = 0.0048, and P = 0.0035, respectively). Positive correlation between levels of apoA-I, apoA-II, and HDL-C and the number of G alleles was observed in females (P = 0.0120, P = 0.0032, and P = 0.0030), but not males (P > 0.05). Because few studies have evaluated the effect of ESR1 gene polymorphisms on lipid traits in children, we also stratified our sample at the age of 15 years. There was evidence of association between intron 1 single-nucleotide polymorphisms rs9322331 and rs9340799 and apoC-II, and triglycerides (TGs) in youths 15 years and younger. In youths, evidence of association between rs9322331 and rs9340799 and apoC-II was stronger in males (P = 0.0036 and P = 0.0124) than in females (P > 0.05), whereas evidence of association with TG was stronger in females (P = 0.0030 and P = 0.0024) than in males (P > 0.05). These findings suggest that ESR1 variation plays an age- and sex-dependent role in determining plasma lipid and apolipoprotein levels.


Genetic structure and core collection of the World Olive Germplasm Bank of Marrakech: towards the optimised management and use of Mediterranean olive genetic resources.
The conservation of cultivated plants in ex-situ collections is essential for the optimal management and use of their genetic resources. For the olive tree, two world germplasm banks (OWGB) are presently established, in Córdoba (Spain) and Marrakech (Morocco). This latter was recently founded and includes 561 accessions from 14 Mediterranean countries. Using 12 nuclear microsatellites (SSRs) and three chloroplast DNA markers, this collection was characterised to examine the structure of the genetic diversity and propose a set of olive accessions encompassing the whole Mediterranean allelic diversity range. We identified 505 SSR profiles based on a total of 210 alleles. Based on these markers, the genetic diversity was similar to that of cultivars and wild olives which were previously characterised in another study indicating that OWGB Marrakech is representative of Mediterranean olive germplasm. Using a model-based Bayesian clustering method and principal components analysis, this OWGB was structured into three main gene pools corresponding to eastern, central and western parts of the Mediterranean Basin. We proposed 10 cores of 67 accessions capturing all detected alleles and 10 cores of 58 accessions capturing the 186 alleles observed more than once. In each of the 10 cores, a set of 40 accessions was identical, whereas the remaining accessions were different, indicating the need to include complementary criteria such as phenotypic adaptive and agronomic traits. Our study generated a molecular database for the entire OWGB Marrakech that may be used to optimise a strategy for the management of olive genetic resources and their use for subsequent genetic and genomic olive breeding.


Innovative use of a standardized debriefing guide to assist in the development of a research questionnaire with low literacy demands.
Prevalence and impact of low literacy has resulted in greater attention to developing written materials at lower reading levels for both patient education and research. The purpose of this study was to develop and evaluate how well a research questionnaire about self-management preferences, intentionally developed as a tool for individuals with low literacy skills, performed. The investigators created a standardized debriefing guide to evaluate comprehension and ease of instrument completion to accompany the administration of the Conventional and Alternative Management for Asthma (CAMA) instrument. The use of a standardized debriefing guide following cognitive interviewing techniques, allowed for the identification of problematic words, unclear meanings and confusion over scaling despite a deliberate attempt to develop a tool with low literacy demands. Such approaches might be considered critically important to insure the accuracy of patient-reported outcomes when self-administered tools are used to collect research and clinical data.


A novel bifunctional GH51 exo-α-l-arabinofuranosidase/endo-xylanase from Alicyclobacillus sp. A4 with significant biomass-degrading capacity.
Improving the hydrolytic performance of xylanolytic enzymes on arabinoxylan is of importance in the ethanol fermentation industry. Supplementation of debranching (arabinofuranosidase) and depolymerizing (xylanase) enzymes is a way to address the problem. In the present study, we identified a bifunctional α-l-arabinofuranosidase/endo-xylanase (Ac-Abf51A) of glycoside hydrolase family 51 in Alicyclobacillus sp. strain A4. Its biochemical stability and great hydrolysis efficiency against complex biomass make it a potential candidate for the production of biofuels. The gene encoding Ac-Abf51A was cloned. The comparison of its sequence with reference proteins having resolved 3D-structures revealed nine key residues involved in catalysis and substrate-binding interaction. Recombinant Ac-Abf51A produced in Escherichia coli showed optimal activity at pH 6.0 and 60 °C with 4-nitrophenyl-α-l-arabinofuranoside as the substrate. The enzyme exhibited an exo-type mode of action on polyarabinosides by catalyzing the cleavage of α-1,2- and α-1,3-linked arabinofuranose side chains in sugar beet arabinan and water-soluble wheat arabinoxylan and α-1,5-linked arabinofuranosidic bonds in debranched sugar beet arabinan. Surprisingly, it had capacity to release xylobiose and xylotriose from wheat arabinoxylan and was active on xylooligosaccharides (xylohexaose 1.2/mM/min, xylopentaose 6.9/mM/min, and xylotetraose 19.7/mM/min), however a lower level of activity. Moreover, Ac-Abf51A showed greater synergistic effect in combination with xylanase (2.92-fold) on wheat arabinoxylan degradation than other reported enzymes, for the amounts of arabinose, xylose, and xylobiose were all increased in comparison to that by the enzymes acting individually. This study for the first time reports a GH51 enzyme with both exo-α-l-arabinofuranosidase and endo-xylanase activities. It was stable over a broad pH range and at high temperature, and showed greater synergistic effect with xylanase on the degradation of wheat arabinoxylan than other counterparts. The distinguished synergy might be ascribed to its bifunctional α-l-arabinofuranosidase/xylanase activity, which may represent a possible way to degrade biomass at lower enzyme loadings.


Risk of biochemical recurrence and timing of radiotherapy in pT3a N0 prostate cancer with positive surgical margin : A single center experience.
Despite improved biochemical recurrence-free survival rates by the use of immediate adjuvant radiotherapy (RT) in patients with locally advanced prostate cancer, disagreement about the need and timing of RT remains. From 2005-2009, 94 patients presenting with a stage pT3a N0 and microscopic positive resection margin were retrospectively analyzed after radical prostatectomy. Special attention was given to patients' outcome, the frequency of additive RT, and its efficacy. Median follow-up was 80 months. A total of 71 patients had a negative postoperative prostate-specific antigen (PSA) level (<0.07 ng/ml). Thirty-six of them did not experience any PSA relapse (subgroup 1). Fourteen of them received additive RT and during follow-up all 36 patients remained PSA negative. Of 71 initially PSA-negative patients, 35 had a biochemical relapse (subgroup 2); 28 patients underwent salvage RT. The median PSA value before salvage RT was 0.24 ng/ml and was subsequently negative (<0.07 ng/ml) in 23 patients after RT. Of the entire cohort, 23 patients had persisting PSA after surgery (subgroup 3). Of these, 18 patients received salvage RT at a median PSA level of 0.4 ng/ml. One patient in subgroup 1, 5 patients in subgroup 2, and 9 patients in subgroup 3 had ongoing androgen deprivation therapy. The present study of 94 pT3a N0 R1 prostate cancer patients provides insight into medical care of this patient cohort and underlines the need for additive RT for the majority of patients to achieve long-term biochemical control. Although immediate adjuvant RT was applied with restraint (20 %), during the observation period 60 of 94 patients (63.8 %) received RT - highlighting the need of postoperative treatment.


Clinical and morphological aspects of nephrotic syndrome in perimembranous, focally sclerosing and membrano-proliferative glomerulonephritis.
The following data was obtained by morphometric and photometric microscopic studies of renal biopsies from 140 patients with idiopathic perimembranous glomerulonephritis, 108 patients with focally sclerosing glomerulonephritis and 50 patients with membranoproliferative glomerulonephritis and correlation of the results obtained with available clinical data: 1. In all three diseases proteinuria and serum protein concentration show no tendency to stabilization in spite of increasing renal insufficiency. 2. With increasing renal insufficiency the proximal tubular cells become increasingly atrophic. 3. Protein resorption through the proximal tubulus cells becomes increasingly diminished with advancing renal insufficiency. It is concluded from the present data that proteinuria, which is primarily glomerular caused, is increased by increasingly diminished resorption of proteins in the tubulus system with increasing renal insufficiency. In this way, even under conditions of advanced renal insufficiency with reduced GFR, large amounts of proteins can be excreted and a nephrotic syndrome can persist to a stage of renal insufficiency.


The effect of DDT and its metabolite (DDE) on prostaglandin secretion from epithelial cells and on contractions of the smooth muscle of the bovine oviduct in vitro.
The insecticide DDT and its metabolite (DDE), due to their lipolytic nature and resistance to biodegradation, are accumulated in the living tissues. In cows, DDT and DDE were found to affect prostaglandin (PG) secretion from the endometrium and contractions of the myometrium. In this study, the impact of both xenobiotics (0.1, 1, 10 or 100ng/ml) on the function of epithelial cells and muscle strips of bovine oviducts from 1 to 5day of the oestrous cycle was examined. Therefore the concentration of PGE2 and PGFM (a metabolite of PGF2α) in culture media, mRNA expression of genes involved in PGs synthesis in epithelial cells and the force and amplitude of strips contractions were measured after 2 and 24 or 48h of incubation. Neither DDT nor DDE affected the viability of cells after 48h (P>0.05). Both DDT and DDE increased the concentrations of PGFM in culture medium and secretion of PGE2 after only 2h of cell culture (P<0.05). Similar effects were seen for the influence of DDE on amount of PGFM after 48h, while DDT decreased secretion of PGE2 (P<0.05). DDT after 2h increased (P<0.05) mRNA expression of PGF2α synthase (PGFS), while both xenobiotics decreased (P<0.05) mRNA expression of cyclooxygenase-2 (COX-2) after 24h. DTT also increased the force of isthmus contractions after 2h, as did both xenobiotics after 48h (P<0.05). Moreover, after 2 and 48h, DDE stimulated the amplitude of contractions of the isthmus as well as the ampulla, (P<0.05). The effect of both compounds on oviduct contractions was diminished by indomethacin, which blocks PG synthesis. We conclude that oviductal secretion of prostaglandins is affected, by DDT and DDE. The influence of these xenobiotics on PGF2α and PGE2 secretion and ratio may be part of the mechanism by which both DDT and its metabolite disturb the contractions of oviductal muscle.


Relationship between adiposity and pedometer-assessed ambulatory activity in adult, rural African women.
To investigate the association between adiposity and pedometry-assessed ambulation in a convenience sample of adult, rural black South African women. Pedometry data were collected over 7 days in 121 subjects. Adiposity measures included body mass index (BMI), waist circumference (WC) and percentage body fat (PBF). Sedentarism (<5000 steps day(-1)) was found in 13.7%, while 39.7% were classified as accruing sufficient physical activity (>or=10 000 steps day(-1)). Significant associations (P<0.02) existed between steps day(-1) and adiposity measures (r=-0.22 to -0.23). After adjusting for age, only BMI remained significantly associated with steps day(-1) (r=-0.20, P=0.032). Significant age-adjusted linear trends were found across combined BMI-WC risk categories for steps day(-1) (P=0.036). Adjusting for age, motor vehicle access, education, use of tobacco products and comorbidities, BMI decreased 1.4 kg m(-2) per 5000 steps day(-1) (P=0.035), access to a motor vehicle within the household increased PBF by 4% (P=0.018), and compared with sedentarism, the risk of obesity (BMI >or=30 kg m(-2)) was 52% lower at 10 000 steps day(-1) (P=0.028). Modest associations were found between adiposity and ambulation. Ambulation decreased the risk for obesity, while motor vehicle access was associated with increased adiposity levels.


The carboxyl terminus of interferon-gamma contains a functional polybasic nuclear localization sequence.
Cytokines such as interferon-gamma (IFN-gamma), which utilize the well studied JAK/STAT pathway for nuclear signal transduction, are themselves translocated to the nucleus. The exact mechanism for the nuclear import of IFN-gamma or the functional role of the nuclear translocation of ligand in signal transduction is unknown. We show in this study that nuclear localization of IFN-gamma is driven by a simple polybasic nuclear localization sequence (NLS) in its COOH terminus, as verified by its ability to specify nuclear import of a heterologous protein allophycocyanin (APC) in standard import assays in digitonin-permeabilized cells. Similar to other nuclear import signals, we show that a peptide representing amino acids 95-132 of IFN-gamma (IFN-gamma(95-132)) containing the polybasic sequence 126RKRKRSR132 was capable of specifying nuclear uptake of the autofluorescent protein, APC, in an energy-dependent fashion that required both ATP and GTP. Nuclear import was abolished when the above polybasic sequence was deleted. Moreover, deletions immediately NH2-terminal of this sequence did not affect the nuclear import. Thus, the sequence 126RKRKRSR132 is necessary and sufficient for nuclear localization. Furthermore, nuclear import was strongly blocked by competition with the cognate peptide IFN-gamma(95-132) but not the peptide IFN-gamma(95-125), which is deleted in the polybasic sequence, further confirming that the NLS properties were contained in this sequence. A peptide containing the prototypical polybasic NLS sequence of the SV40 large T-antigen was also able to inhibit the nuclear import mediated by IFN-gamma(95-132). This observation suggests that the NLS in IFN-gamma may function through the components of the Ran/importin pathway utilized by the SV40 T-NLS. Finally, we show that intact IFN-gamma, when coupled to APC, was also able to mediate its nuclear import. Again, nuclear import was blocked by the peptide IFN-gamma(95-132) and the SV40 T-NLS peptide, suggesting that intact IFN-gamma was also transported into the nucleus through the Ran/importin pathway. Previous studies have suggested a direct intracellular role for IFN-gamma in the induction of its biological activities. Based on our data in this study, we suggest that a key intracellular site of interaction of IFN-gamma is the one with the nuclear transport mechanism that occurs via the NLS in the COOH terminus of IFN-gamma.


Localization of the binding site for cell attachment in the alpha1(I) chain of collagen.
Certain cells such as CHO (Chinese hamster ovary) and fibroblasts attach to a substrate of type I collagen via proteins that link the cell surface to collagen. Serum contains a glycoprotein, c-CAP (collagen cell attachment protein), that can mediate this adhesion. Previously, we established that alpha1(I)-CB7 (cyanogen bromide peptide 7), which lies within residues 552 to 822 of collagen, contains a major binding site for c-CAP(alpha1(I)-CB7 has been renumbered to account for an additional triplet recently identified at residue 613 (P. P. Fietzek and R. W. Glanville, manuscript in preparation). Now we have examined the ability of various peptides derived by proteolytic digestion of alpha1(I)-CB7 to bind to c-CAP based upon their ability to inhibit cell attachment to collagen. The binding site lies within residues 757 to 791. It is likely that this is the sole binding site in the alpha1(I) chain since cleavage of the bond between residues 775 and 776 in the alpha1(I) chain destroys cell attachment activity.


Differential diagnosis of seizures.
Distinguishing epileptic events from nonepileptic paroxysmal neurologic events represents a common diagnostic challenge. Syncope, either cardiac or noncardiac, can appear similar to atonic and even convulsive seizures. Breath holding and benign paroxysmal vertigo in children may be confused with epilepsy. Classic migraine, transient global amnesia, and transient ischemic attacks may resemble epileptic seizures. Sleep disorders, including nocturnal movements, parasomnias, and narcolepsy also may resemble epileptic seizures. Most movement disorders are distinguished easily from epilepsy; however, paroxysmal dyskinesias may resemble atonic or reflex seizures. The correct diagnosis can be established and appropriate treatment can be instituted by relying on routine and prolonged EEG, EKG, and sleep studies, when appropriate.


Significance of CNS 2 cerebrospinal fluid status post-induction in pediatric and adolescent patients with acute lymphoblastic leukemia.
At diagnosis, there are prognostic implications of low-level leukemic blasts (CNS 2) in the cerebrospinal fluid (CSF) of patients with acute lymphoblastic leukemia (ALL). However, the significance of post-induction CNS 2 results and the impact of equipment on CNS 2 prevalence have not been well studied. A single-institution retrospective cohort study was conducted to analyze the outcome of patients with ≥1 post-induction CNS 2. A subanalysis compared the proportion of CNS 2 CSF results using 2 different cytocentrifuges; the Shandon Cytospin used from 2005 to 2008 and the Wescor Cytopro used from 2010 to 2014. Over 4500 post-induction CSF samples were analyzed, of which 59 were CNS 2. In covariate-adjusted analyses, post-induction CNS 2 did not significantly increase relapse risk. The proportion of CNS 2 results increased 4.3-fold in noninfants and 6.3-fold in infants using the Wescor Cytopro. Cytocentrifuge machine did not affect CNS 3 prevalence. These findings support our current practice of not changing management based on a post-induction CNS 2 CSF and highlight how equipment changes can significantly influence testing results. More data are needed to analyze relapse by subpopulations, such as those with repeated CNS 2 findings.


Retinal vascular calibre and the risk of coronary heart disease-related death.
To examine whether retinal vascular calibre independently predicts risk of coronary heart disease (CHD) -related death. In a population-based cohort study of 3654 Australians aged > or = 49 years, retinal arteriolar and venular calibres were measured from baseline retinal photographs and the arteriole to venule ratio (AVR) was calculated. CHD-related death was confirmed from the Australian National Death Index. Over nine years, 78 women (4.1%) and 114 men (7.8%) had incident CHD-related deaths. In people aged 49-75 years, wider venules were associated with CHD death, with relative risk (RR) 1.8 (95% confidence interval (CI) 1.1 to 2.7) and RR 2.0 (95% CI 1.1 to 3.6) per standard deviation (SD) increase in venular calibre for men and women, respectively, after adjustment for traditional risk factors. Additionally, in women aged 49-75 years, smaller AVR and narrower arterioles were associated with CHD death (RR 1.5, 95% CI 1.1 to 2.2, and RR 1.9, 95% CI 1.0 to 3.5 per SD decrease in AVR and arteriolar calibre, respectively, after adjustment). These associations were not observed in people aged > 75 years. These findings suggest that microvascular disease processes may have a role in CHD development in middle-aged people, particularly in women. Retinal photography may be useful in cardiovascular risk prediction.


Simultaneous determination of hydroxocobalamin and its cyanide complex cyanocobalamin in human plasma by high-performance liquid chromatography. Application to pharmacokinetic studies after high-dose hydroxocobalamin as an antidote for severe cyanide poisoning.
Hydroxocobalamin (OHCbl) is a powerful antidote for cyanide poisoning, via the formation of non-toxic cyanocobalamin (CNCbl). Plasmatic cobalamins were measured at 361 nm, after enrichment and purification on a short C18 precolumn (1% acetic acid; 1 ml min-1; 2 min), by back-flush elution on a C18 ODS-2 column [0.1 M sodium dihydrogenphosphate-methanol (63:27, v/v) (pH 4.0); 0.80 ml min-1]. The precision was 3.21 and 3.54% for 10 microM OHCbl and CNCbl, respectively. The method was used to study the pharmacokinetics of OHCbl and the formed CNCbl in severely poisoned patients.


Characterization of the novel HLA-DRB1*12:82 allele by sequencing-based typing.
HLA-DRB1*12:82 differs from HLA-DRB1*12:01:01:01 by one nucleotide substitution in codon 104 in exon 3.


Reversible mental deterioration and neurological disturbances with o,p'-DDD therapy.
A patient with serious neurological and neuropsychological side-effects of a 2-year treatment with 1,1-dichloro-(o-chlorophenyl)-2-(p-chlorophenyl)ethane (o,p'-DDD) for adrenal carcinoma showed complete recovery after discontinuation of the drug. Studying a larger number of patients during and after long term treatment with o,p'-DDD is indicated. If results of this report are confirmed, therapy should not be discontinued even if serious side effects occur.


Fluoroscopy-guided infiltration for pain reduction in patients with Baastrup's disease: clinical experience and results.
Baastrup disease refers to degenerative changes of adjacent spinous processes with resultant back pain. The purpose of this study is to assess the safety and efficacy of percutaneous, fluoroscopy-guided infiltrations in a consecutive series of patients suffering from Baastrup disease. From January 2009 until December 2013, 55 patients suffering from Baastrup disease (diagnosed clinically and by imaging findings) underwent percutaneous, fluoroscopy-guided infiltration. The position of the needle (22-gauge spinal needle) was fluoroscopically verified at the level of interspinous ligament. Then a mixture of long-acting corticosteroid with local anesthetic (1.5/1 cc) was injected. A questionnaire with NVS scale helped in assessing pain relief degree, life quality, and mobility improvement. In the patients of our study, a total of 67 sessions was performed. In 12/55 patients (22%), a second infiltration was performed within 7-10 days apart from the first one. The end point was 1 year post-treatment. Patients were followed 1 week after the first injection for a subsequent treatment and then if asymptomatic re-controlled in 3-6 and 12 months. Comparing the pain scores prior (mean value, 8.18 ± 1.44 NVS units) and after (mean value, 0.62 ± 0.93 NVS units) there was a mean decrease of 7.56 ± 1.68 NVS units (p < 0.001) on terms of pain reduction, effect upon mobility and life quality. There were no clinically significant complications noted in our study. Fluoroscopy-guided infiltrations seem to be a feasible, efficacious, and safe approach for pain reduction and mobility improvement in patients with Baastrup disease.


Immediate-early gene expression in the central nucleus of the amygdala is not specific for anxiolytic or anxiogenic drugs.
The lateral, basal, and central nuclei of the amygdala are part of a circuitry that instantiates many fear and anxious behaviors. One line of support indicates that immediate-early gene (IEG) expression (e.g., c-fos and egr-1 (zif268)) is increased in these nuclei following fear conditioning. Other research finds that anxiogenic drugs working through various mechanisms induce IEG expression in the central nucleus of the amygdala (CeA) suggesting that expression is a neural marker for fear and anxiety. However, several studies have also found that anxiolytic drugs induce IEG expression in the CeA. Expression of egr-1 in the CeA and lateral nucleus of the amygdala following administration of anxiolytic and anxiogenic benzodiazepine and serotonin agonists and antagonists was investigated. The first experiment determined behaviorally active anxiolytic and anxiogenic doses for two anxiogenic drugs (FG 7142 and mCPP) and two anxiolytic drugs (diazepam and buspirone). The effects of anxiogenic and anxiolytic doses of these drugs on egr-1 expression in the amygdala were then tested in a second experiment. All four drugs increased egr-1 in the CeA indicating that increased egr-1 mRNA expression in the CeA is not specific to anxiolytic or anxiogenic effects of the drugs. We suggest that IEG expression in the CeA may be due to activation of circuits that are associated with systemic physiological homeostasis perturbed by a number of drugs including anxiogenic and anxiolytic compounds.


Ruptured sinus of Valsalva aneurysm associated with aortic regurgitation caused by hemodynamic effect solely.
We describe a case of ruptured sinus of Valsalva aneurysm (RSVA) with moderate aortic regurgitation (AR), which developed on the second day after admission. The AR was caused by a hemodynamic effect solely, in which the shunt blood flow through ruptured site pulled the right aortic cusp away from closure. The pathological mechanism of the AR was clearly visualized by intraoperative transesophageal echocardiography (TEE) and the AR was successfully resolved after simple closure of the RSVA without any additional procedure to the aortic valve.


Risk of second primary cancer among women in the Kaiser Permanente Breast Cancer Survivors Cohort.
Breast cancer survivors are living longer due to early detection and advances in treatment and are at increased risk for second primary cancers. Comprehensive evaluation of second cancer risk among patients treated in recent decades is lacking. We identified 16,004 females diagnosed with a first primary stage I-III breast cancer between 1990 and 2016 (followed through 2017) and survived ≥ 1 year at Kaiser Permanente (KP) Colorado, Northwest, and Washington. Second cancer was defined as an invasive primary cancer diagnosed ≥ 12 months after the first primary breast cancer. Second cancer risk was evaluated for all cancers (excluding ipsilateral breast cancer) using standardized incidence ratios (SIRs), and a competing risk approach for cumulative incidence and hazard ratios (HRs) adjusted for KP center, treatment, age, and year of first cancer diagnosis. Over a median follow-up of 6.2 years, 1,562 women developed second cancer. Breast cancer survivors had a 70% higher risk of any cancer (95%CI = 1.62-1.79) and 45% higher risk of non-breast cancer (95%CI = 1.37-1.54) compared with the general population. SIRs were highest for malignancies of the peritoneum (SIR = 3.44, 95%CI = 1.65-6.33), soft tissue (SIR = 3.32, 95%CI = 2.51-4.30), contralateral breast (SIR = 3.10, 95%CI = 2.82-3.40), and acute myeloid leukemia (SIR = 2.11, 95%CI = 1.18-3.48)/myelodysplastic syndrome (SIR = 3.25, 95%CI = 1.89-5.20). Women also had elevated risks for oral, colon, pancreas, lung, and uterine corpus cancer, melanoma, and non-Hodgkin lymphoma (SIR range = 1.31-1.97). Radiotherapy was associated with increased risk for all second cancers (HR = 1.13, 95%CI = 1.01-1.25) and soft tissue sarcoma (HR = 2.36, 95%CI = 1.17-4.78), chemotherapy with decreased risk for all second cancers (HR = 0.87, 95%CI = 0.78-0.98) and increased myelodysplastic syndrome risk (HR = 3.01, 95%CI = 1.01-8.94), and endocrine therapy with lower contralateral breast cancer risk (HR = 0.48, 95%CI = 0.38-0.60). Approximately 1 in 9 women who survived ≥ 1 year developed second cancer, 1 in 13 developed second non-breast cancer, and 1 in 30 developed contralateral breast cancer by 10 years. Trends in cumulative incidence declined for contralateral breast cancer but not for second non-breast cancers. Elevated risks of second cancer among breast cancer survivors treated in recent decades suggests that heightened surveillance is warranted and continued efforts to reduce second cancers are needed.


Origins of the vagal drive controlling left ventricular contractility.
The strength, functional significance and origins of parasympathetic innervation of the left ventricle remain controversial. This study tested the hypothesis that parasympathetic control of left ventricular contractility is provided by vagal preganglionic neurones of the dorsal motor nucleus (DVMN). Under β-adrenoceptor blockade combined with spinal cord (C1) transection (to remove sympathetic influences), systemic administration of atropine increased left ventricular contractility in rats anaesthetized with urethane, confirming the existence of a tonic inhibitory muscarinic influence on cardiac inotropy. Increased left ventricular contractility in anaesthetized rats was observed when DVMN neurones were silenced. Functional neuroanatomical mapping revealed that vagal preganglionic neurones that have an impact on left ventricular contractility are located in the caudal region of the left DVMN. These neurones provide functionally significant parasympathetic control of left ventricular inotropy. The strength, functional significance and origins of direct parasympathetic innervation of the left ventricle (LV) remain controversial. In the present study we used an anaesthetized rat model to first confirm the presence of tonic inhibitory vagal influence on LV inotropy. Using genetic neuronal targeting and functional neuroanatomical mapping we tested the hypothesis that parasympathetic control of LV contractility is provided by vagal preganglionic neurones located in the dorsal motor nucleus (DVMN). It was found that under systemic β-adrenoceptor blockade (atenolol) combined with spinal cord (C1) transection (to remove sympathetic influences), intravenous administration of atropine increases LV contractility in rats anaesthetized with urethane, but not in animals anaesthetized with pentobarbital. Increased LV contractility in rats anaesthetized with urethane was also observed when DVMN neurones targeted bilaterally to express an inhibitory Drosophila allatostatin receptor were silenced by application of an insect peptide allatostatin. Microinjections of glutamate and muscimol to activate or inhibit neuronal cell bodies in distinct locations along the rostro-caudal extent of the left and right DVMN revealed that vagal preganglionic neurones, which have an impact on LV contractility, are located in the caudal region of the left DVMN. Changes in LV contractility were only observed when this subpopulation of DVMN neurones was activated or inhibited. These data confirm the existence of a tonic inhibitory muscarinic influence on LV contractility. Activity of a subpopulation of DVMN neurones provides functionally significant parasympathetic control of LV contractile function.


Spatial distribution of surface action potentials generated by individual motor units in the human biceps brachii muscle.
This study analyses the spatial distribution of individual motor unit potentials (MUPs) over the skin surface and the influence of motor unit depth and recording configuration on this distribution. Multichannel surface (13×5 electrode grid) and intramuscular (wire electrodes inserted with needles of lengths 15 and 25mm) electromyographic (EMG) signals were concurrently recorded with monopolar derivations from the biceps brachii muscle of 10 healthy subjects during 60-s isometric contractions at 20% of the maximum torque. Multichannel monopolar MUPs of the target motor unit were obtained by spike-triggered averaging of the surface EMG. Amplitude and frequency characteristics of monopolar and bipolar MUPs were calculated for locations along the fibers' direction (longitudinal), and along the direction perpendicular (transverse) to the fibers. In the longitudinal direction, monopolar and bipolar MUPs exhibited marked amplitude changes that extended for 16-32mm and 16-24mm over the innervation and tendon zones, respectively. The variation of monopolar and bipolar MUP characteristics was not symmetrical about the innervation zone. Motor unit depth had a considerable influence on the relative longitudinal variation of amplitude for monopolar MUPs, but not for bipolar MUPs. The transverse extension of bipolar MUPs ranged between 24 and 32mm, whereas that of monopolar MUPs ranged between 72 and 96mm. The mean power spectral frequency of surface MUPs was highly dependent on the transverse electrode location but not on depth. This study provides a basis for the interpretation of the contribution of individual motor units to the interference surface EMG signal.


[Progress in the studies of vasoactive intestinal polypeptide].
Vasoactive intestinal polypeptide (VIP) is a potent vasodilator and smooth muscle relaxant, and also fulfils several of the criteria for a neurotransmitter mediating penile erection. VIP donor and gene transfer of VIP are a potentially promising and physiologically relevant strategies for the treatment of erectile dysfunction.


Evaluation of Impact of Interferon-Induced Helicase C Domain-Containing Protein 1 Gene in Egyptian Systemic Lupus Erythematosus Patients and its Relationship With Vascular Manifestations of the Disease.
This study aims to investigate the impact of interferon-induced helicase C domain-containing protein 1 (IFIH1) gene single nucleotide polymorphism on interferon pathway signaling in systemic lupus erythematosus (SLE) patients specifically with vascular affection. The cross-sectional study included 30 consecutive SLE patients (2 males, 28 females; mean age 28±3.4 years; range 16 to 40 years) diagnosed according to the American College of Rheumatology revised criteria and 10 healthy age- and sex-matched controls (2 males, 8 females; mean age 27±2.5 years; range 22 to 23 years). SLE patients and controls were compared in terms of quantitative reverse transcriptase polymerase chain reaction gene expression of IFIH1 gene, von Willebrand factor, carotid intima-media thickness, and ankle brachial index. Interferon-induced helicase C domain-containing protein 1 gene expression was significantly higher in SLE patients than controls (1.7±0.6 and 0.5±0.2, respectively) (p<0.0001). IFIH1 gene expression was highly related to vascular complication with a cutoff point at 1.74 and it positively correlated with other endothelial dysfunction markers. Interferon-induced helicase C domain-containing protein 1 gene (single nucleotide polymorphism 1990670) is associated with SLE in Egyptian patients. Expression of IFIH1 gene is related to disease activity and may serve as a novel predictor of vascular affection in SLE.


Is diabetes mellitus a major risk factor in coronary artery bypass grafting? The influence of internal thoracic artery grafting on late survival in diabetic patients.
The presence of diabetes could possibly have an adverse influence on the late results of coronary artery bypass grafting (CABG). A retrospective observational risk-unadjusted estimation study was conducted to clarify the magnitude of this unfavorable influence of diabetes based on our experience at Juntendo University. Between January 1984 and December 1995, 1,618 primary CABG operations were performed and 99.0% of these patients were followed up for a mean period of 81.2 months with a maximal follow-up of 14 years. Among the 1,610 patients who were successfully followed up, 523 patients (32.5%) were diabetic and this cohort was compared with 1,087 nondiabetic patients regarding the patient profiles, perioperative outcome, and the long-term outcome. Diabetic patients included a higher proportion of females, a greater preoperative presence of renal insufficiency, a higher rate of three vessel disease, and reception of a greater number of grafts (mean: 2.5 versus 2.4) and an internal thoracic artery graft (68% versus 58%). Hospital mortality was not significantly different (2.1% vs 1.0%: ns). An actuarial analysis revealed a significantly unfavorable long-term survival in the diabetic group (59.6%) at 14 years, compared with the nondiabetic group (73.4%), however diabetic patients with an internal thoracic artery graft had a favorable long-term survival (82.0%) at 13 years, and this finding was almost identical to that of nondiabetic patients with an internal thoracic artery graft (88.5%). We conclude that the presence of diabetes appears to be a major risk factor in patients undergoing CABG. However, diabetic patients with an internal thoracic artery graft had a survival almost identical to that of nondiabetic patients with an internal thoracic artery graft.


What young men know about testicular cancer.
A survey to asess what young men know about testicular cancer and to determine awareness and practice of testicular self-examination (TSE) was conducted among 266 male graduate and undergraduate students at the State University of New York at Buffalo. Findings indicate that the majority of young men were either uninformed or misinformed about their risk of getting testicular cancer and about the symptoms of testicular cancer. Only 16% of respondents had heard of TSE before the survey, and only a small percentage of these individuals reported doing TSE on a routine basis. The findings from this study point out the need for increased public education directed at symptom recognition, especially at the high school and post-high school levels.


Predictors of Caries Risk among Egyptian Children Attending Pediatric Dental Clinics at a University Hospital.
Dental caries is the most prevalent dental disease. The external validity of the available caries risk assessment (CRA) tools is not established, especially among pediatric population. To assess caries risk using the caries management by risk assessment (CAMBRA) protocol among Egyptian children aged 3-12 years and suggest variables that could potentially be used to develop a simpler CRA model. For this cross-sectional study, we recruited 320 children aged 3 to <6 years (Group I) and 320 children aged 6-12 years (Group II). CAMBRA was used to collect data about disease indicators, biological and environmental factors, and protective factors among study participants. Each child was examined clinically to collect data about past caries experiences and to measure plaque scores. The risk of caries was high in 92.5% of Group I and 83.4% of Group II participants. The overall dmft was 5.71 ± 3.18 for Group I and 4.78 ± 2.53 for Group II. In Group I, a significant positive relation was found between the overall mean caries risk score and past caries experience (dmft; r = 0.344, P < 0.001) and mean plaque index (r = 0.463, P < 0.001). In Group II, a significant positive relation was found between the overall mean caries risk score and dmft score (r = 0.511, P < 0.001), S. mutans count (r = 234, P < 0.001), Lactobacilli count (r = 0.316, P < 0.001), and plaque index (r = 0.463, P < 0.001). Participants' age, parents' education, and parents' occupation had a negative significant effect on the overall mean caries risk score. This study suggests predictors that can be used in the development of a new CRA model for children aged 3-12 years.


Audio-visual integration during speech perception in prelingually deafened Japanese children revealed by the McGurk effect.
To investigate the McGurk effect in profoundly deafened Japanese children with cochlear implants (CI) and in normal-hearing children. This was done to identify how children with profound deafness using CI established audiovisual integration during the speech acquisition period. Twenty-four prelingually deafened children with CI and 12 age-matched normal-hearing children participated in this study. Responses to audiovisual stimuli were compared between deafened and normal-hearing controls. Additionally, responses of the children with CI younger than 6 years of age were compared with those of the children with CI at least 6 years of age at the time of the test. Responses to stimuli combining auditory labials and visual non-labials were significantly different between deafened children with CI and normal-hearing controls (p<0.05). Additionally, the McGurk effect tended to be more induced in deafened children older than 6 years of age than in their younger counterparts. The McGurk effect was more significantly induced in prelingually deafened Japanese children with CI than in normal-hearing, age-matched Japanese children. Despite having good speech-perception skills and auditory input through their CI, from early childhood, deafened children may use more visual information in speech perception than normal-hearing children. As children using CI need to communicate based on insufficient speech signals coded by CI, additional activities of higher-order brain function may be necessary to compensate for the incomplete auditory input. This study provided information on the influence of deafness on the development of audiovisual integration related to speech, which could contribute to our further understanding of the strategies used in spoken language communication by prelingually deafened children.


Catalysis of the disproportionation of superoxide by metalloporphyrins.
The efficiencies of several metalloporphyrin complexes at catalyzing the disproportionation of superoxide have been determined at pH 10 in both carbonate and borate buffer systems. Catalytic rate constants were obtained for the iron(III) and cobalt(III) derivatives of tetrakis(4-N-methylpyridyl) porphine, for tetraphenylporphinesulfonatoferrate(III) and for hemin. In addition, the effects of added bovine serum albumin and imidazole were studied. The order of catalytic efficiency is FeTMpyP greater than FeTMpyP(Im)2 greater than FeTPPS(Im)2 approximately FeTPPS approximately FeTPPS.BSA approximately Fe(EDTA) greater than or approximately CoTMpyP greater than hemin(Im)2 greater than or approximately hemin.


Differential diagnosis of detrusor instability and stress-incontinence by patient history: the Gaudenz-Incontinence-Questionnaire revisited.
To evaluate the validity of the Gaudenz-Incontinence-Questionnaire in the differential diagnosis of genuine stress-incontinence and detrusor instability. Diagnoses based on questionnaire-results were compared to those following complete urogynecologic assessment including urodynamics in 1938 patients with lower urinary tract symptoms. Sensitivity and specificity for diagnosis of stress-incontinence were 0.559 and 0.447, respectively, for detrusor instability 0.615 and 0.563, respectively. The low sensitivity and specificity of the test do not justify its use as a diagnostic tool in patients with urinary incontinence. These results show that the scores may be misleading in a large proportion of patients and should not therefore be the only determinant of diagnosis, nor should strategies be based on history alone.


The effect of aldosterone on adiposity - The role of glucose absorption in the small intestine.
We have previously demonstrated that manipulation of the renin angiotensin system (RAS) has large effects on digestive efficiency. However, the effects of aldosterone on body weight, adiposity, and glucose absorption in the intestine remains unknown. We here demonstrated that lack of aldosterone synthase (ASKO) in mice did not affect adiposity. In contrast, mice administered with aldosterone were resistant to diet-induced obesity. This is due to gastrointestinal loss of dietary glucose. As expected, ASKO mice had increased glucose absorption, whereas mice administered with aldosterone had reduced glucose absorption in the small intestine. Furthermore, the level of protein expression of sodium glucose transporter 1 (SGLT1) in the mucosa of the jejunum was higher in ASKO mice, and lower in mice administered with aldosterone than control mice. Our findings indicate that aldosterone plays an important role on SGLT-1-mediated glucose absorption in the small intestine.


Novel exosomal miR-46146 transfer oxaliplatin chemoresistance in colorectal cancer.
Oxaliplatin (OX) is widely used for patients with advanced colorectal cancer (CRC). However, most of them will turn out to be OX resistant. Therefore, it is necessary to uncover the causes underlying this phenomenon. Emerging works have reported that exosomal miRNAs are linked to chemoresistance in many types of cancer. Hence, we separated exosomes from OX sensitive (Exo-S) and resistant CRC cells (Exo-R) by ultracentrifugation and characterized those exosomes by transmission electron microscope and Nanosight NS300. The differentiated miRNAs between Exo-S and Exo-R were identified by small RNA deep sequencing. The expression of miRNA was examined by quantitative real-time PCR (qRT-PCR). The effect of Exo-R and exosomal miR-46146 was determined by CCK-8 assay and flow cytometry (FCM). The target gene of miR-46146 was predicted by computational algorithms and validated by dual luciferase assay. We found that parental OX sensitive CRC cell acquired increased resistance to the cytotoxicity of OX when they were cocultured with exosomes secreted by OX-resistant CRC. Notably, a novel miRNA miR-46146 was identified and proved to be upregulated in the Exo-R which was internalized by its recipient cells and contributes to the chemoresistance transfer. Furthermore, we demonstrated that PDCD10 was the direct functional target of miR46146 and augmentation of the PDCD10 expression might reverse the effect of Exo-miR-46146-driven chemoresistance. These results indicate that exosomal miR-46146 functions as a vital mediator of OX resistance by targeting PDCD10 and could be a potential target to re-sensitize CRC cell to OX.


Characterization of isoforms and genomic organization of mouse calumenin.
Calumenin is a multiple EF-hand protein located in endo/sarcoplasmic reticulum of mammalian heart and other tissues [J. Biol. Chem. 272 (1997) 18232; Genomics 49 (1998) 331; Biochim. Biophys. Acta 1386 (1998) 121]. In the present study, a new isoform of mouse calumenin (mouse calumenin 2) was cloned by RT-PCR and genomic DNA PCR. The deduced amino acid sequence of mouse calumenin 2 is 315 aa long with the calculated MW of 37,064 and pI of 4.26. It has 92% aa sequence identity to previously identified mouse calumenin [J. Biol. Chem. 272 (1997) 18232] (mouse calumenin 1). The difference in the aa sequence was restricted to the first two EF-hand regions (residues 74-138). Northern blot analysis shows that mouse calumenin 2 is highly expressed in heart, lung, testis and unpregnant uterus. The expression of mouse calumenin 2 appears to decrease when fetal development is progressed. Genomic DNA PCR, sequencing and data mining of mouse genome database were utilized to examine the exon-intron boundaries of mouse calumenin genes. Both mouse calumenin 1 and 2 genes encompass six exons, and five of them (Exon1, 3, 4, 5 and 6) are identical. However, mouse calumenin 1 contains Exon2-1, whereas mouse calumenin 2 contains a neighboring Exon2-2. The calumenin genes are localized on mouse chromosome 6 having conserved synteny with human chromosome 7q32. For comparison, the genomic organization of human calumenin was also examined using the published human genome database (UCSC Genome Bioinformatics at ). Like mouse calumenin genes, two human calumenin genes also consist of five identical exons (Exon1, 3, 4, 5 and 6) and a different Exon2. The present study suggests that the genomic organization of calumenin genes is well conserved between human and mouse.


Effects of pre-learning stress on memory for neutral, positive and negative words: Different roles of cortisol and autonomic arousal.
Stress can have enhancing or impairing effects on memory. Here, we addressed the effect of pre-learning stress on subsequent memory and asked whether neutral and emotionally valent information are differentially affected by specific stress components, autonomic arousal and stress-induced cortisol. Ninety-six healthy men and women underwent either a stressor (modified cold pressor test) or a control warm water exposure. During stress, participants showed comparable autonomic arousal (heart rate, blood pressure), while 60 percent showed an increase of cortisol (responders vs. 40 percent non-responders). Ten minutes after the cold pressor test neutral, positive and negative words were presented. Free recall was tested 1 and 24h later. Overall, positive and negative words were better recalled than neutral words. Stress enhanced the recall of neutral words independently of cortisol response. In contrast, the free recall of negative words was enhanced in cortisol responders in the 1-h but not 24-h test which might suggest different effects of cortisol on consolidation and reconsolidation processes. Recall for positive words was unaffected by stress-induced cortisol. To summarize, (i) pre-learning stress can enhance memory for neutral words independently of cortisol and (ii) stress effects on memory for negative words appear to rely on stress-induced cortisol elevations, the absence of this effect for positive words might be at least partly due to differences in arousal evoked by positive vs. negative words.


Horseshoe adrenal gland associated with spinal and vascular anomalies.
Horseshoe (or butterfly) adrenal gland (HAG) is an exceedingly rare congenital abnormality compared to horseshoe kidney. Here we present computed tomography findings of HAG in a 70-year-old man, associated with some spinal and vascular abnormalities; in addition, a brief review of HAG in adult people is presented. In our case, spinal anomalies included a severe scoliosis, a "butterfly" deformation of Th9 vertebra and the presence of a "split lumbosacral thecal sac" at L5 level. As regards, vascular anomalies, they were noted: the origin of the left common carotid artery from the innominate artery and the absence of celiac trunk with separate origin of three branches directly off the aorta (a splenic artery, a left gastrohepatic trunk and a right hepatic-pancreaticoduodenal artery).


The prognostic study of mental stress-induced myocardial ischemia in coronary revascularization patients with depression/anxiety: rationale and design.
Mental stress-induced myocardial ischemia (MSIMI) frequently occurs in patients with coronary artery disease (CAD), and is even more common in patients with co-occurring CAD and depression/anxiety. MSIMI appears to be a poor prognostic factor for CAD, but existing data on depression/anxiety patients are limited. This cohort study will consecutively screen 2,647 CAD patients between 2023 and 2025. Included subjects will need to have received coronary revascularization and also have depression and/or anxiety at baseline. This study will enroll 360 subjects who meet the criteria. Two mental stress tests will be carried out in each patient at 1 month and 1 year timelines after coronary revascularization, using Stroop color word tests. MSIMI will be assessed by 99 m-Tc-sestamibi myocardial perfusion imaging. The endothelial function will be assessed by EndoPAT. Furthermore, we will dynamically monitor patients' health and mental conditions every 3 months. The mean follow-up time will be 1 year. The primary endpoint is the major adverse cardiac events, a composite of all-cause death, cardiac death, myocardial infarction, stroke, or unplanned revascularization. Secondary endpoints will include overall health and mental conditions. The reproducibility of mental stress combined with myocardial perfusion for detecting MSIMI and comparisons between coronary stenosis and ischemic segments will also be included. This cohort study will provide information on MSIMI outcomes in CAD patients who also have comorbid depression/anxiety after revascularization. In addition, understanding the long-term dynamics of MSIMI and the match between coronary stenosis and ischemia will provide insight into MSIMI mechanisms. ChiCTR2200055792, 2022.1.20, www.medresman.org.cn.


Comparative study of photocatalytic activities of hydrothermally grown ZnO nanorod on Si(001) wafer and FTO glass substrates.
ZnO nanorods have been grown on Si(001) wafer and fluorine-doped tin oxide (FTO) glass substrates for 1 and 4 h with the hydrothermal methods. The morphologies and photocatalytic activities of the ZnO nanorods were found to depend on the substrates. We investigated their properties by using spectroscopic analysis and demonstrated that the shape of nanorod and the ratios of external defects can be controlled by varying the substrates. Our experiments revealed that the nanorods grown on Si(001) have a single-crystalline wurtzite structure with (002) facets and that the number of surface oxygen defects increases with their length as the growth time increases. The nanorods grown on Si(001) have different facets, in particular wider (002) facets, and a higher ratio of the oxygen defect than the nanorods on FTO glass substrate. Moreover, the photocatalytic activities with respect to 2-aminothiophenol (2-ATP) of these nanorods were investigated with high-resolution photoemission spectroscopy (HRPES). We demonstrated that their photocatalytic activity is influenced by the ratios of surface oxygen defects, which varies with the substrate surface.


Right Ventricular Perforation Leading to Sudden Death Due to Extracorporeal Membrane Oxygenation (ECMO) Canula Dislodgement.
ECMO has been playing an increasingly important role in the management of coronavirus disease (COVID-19)-related acute respiratory distress syndrome (ARDS). However, despite its potential benefits, high mortality rates are still being reported worldwide. Herein, we report the case of a 32-year-old male who presented with worsening shortness of breath secondary to COVID-19. Unfortunately, he experienced a sentinel event when the cannula became dislodged due to coughing, which led to a right ventricular perforation and sudden pulseless electrical activity (PEA) cardiac arrest.


ImmunoPET Imaging of Pancreatic Tumors with 89Zr-Labeled Gold Nanoparticle-Antibody Conjugates.
Targeted delivery in vivo remains an immense roadblock for the translation of nanomaterials into the clinic. The greatest obstacle is the mononuclear phagocyte system (MPS), which sequesters foreign substances from general circulation and causes accumulation in organs such as the liver and spleen. The purpose of this study was to determine whether attaching an active targeting antibody, 5B1, to the surface of gold nanoparticles and using clodronate liposomes to deplete liver and splenic macrophages could help to minimize uptake by MPS organs, increase targeted delivery to CA19.9-positive pancreatic tumors, and enhance pancreatic tumor delineation. To produce the antibody-gold nanoparticle conjugate (Ab-AuNP), the Ab was conjugated to p-isothiocyanatobenzyl-desferrioxamine (p-SCN-DFO) and subsequently conjugated to NHS-activated gold nanoparticles. The Ab-AuNP was characterized by transmission electron microscopy (TEM) and atomic force microscopy (AFM). Modified Lindmo assay was performed to assess binding affinity and internalization potential in vitro. The Ab-AuNP was radiolabeled with 89Zr and injected into CA19.9-positive BxPc-3 pancreatic orthotopic tumor-bearing mice pretreated with or without clodronate liposomes for PET imaging and biodistribution studies. Inductively coupled plasma-optical emission spectrometry (ICP-OES) analysis was used to confirm delivery of gold nanoparticles to BxPc-3 pancreatic subcutaneous xenografts. Mice pretreated with clodronate liposomes in an orthotopic setting demonstrated decreased liver uptake at early time points (12.2 ± 2.3 % ID/g vs. 22.8 ± 3.8 % ID/g at 24 h) and increased tumor uptake at 120 h (13.8 ± 8.0 % ID/g vs. 6.0 ± 1.2 % ID/g). This allowed for delineation of orthotopic pancreatic xenografts in significantly more mice treated with clodronate (6/6) than in mice not treated with clodronate (2/6) or mice injected with gold nanoparticles labeled with a nonspecific antibody (0/5). The combination of clodronate liposomes and an active targeting antibody on the surface of gold nanoparticles allowed for PET/CT imaging of subcutaneous and orthotopic pancreatic xenografts in mice.


The licensing system of Dutch hospitals: a method for quality assurance in health care.
The Dutch licensing system for hospitals is an outstanding example of quality assurance through indirect legislation. In this method the government sets the general frame within which hospitals are free to establish their own procedures. Some requisites for effectiveness and efficiency of a licensing system are formulated. It is argued that if hospitals and health professionals do not fulfil certain requirements when establishing regulatory mechanisms on a voluntary basis, the government will have to intervene with a view to protect the patient.


Effect of Methotrexate on Salpingostomy Completion Rate for Tubal Ectopic Pregnancy: A Retrospective Cohort Study.
To determine whether completion rates of salpingostomy for tubal ectopic pregnancy are compromised by initial medical management with methotrexate (MTX). Retrospective cohort study. Single academic hospital system. Patients requiring surgery for ectopic pregnancy between 2006 and 2017. A subset of patients who went directly to surgery, and all patients who failed MTX before requiring surgery underwent detailed chart review. Salpingostomy plan and success rate and salpingostomy failure reasons were compared between patients pretreated with MTX and those who were MTX-untreated. Among 94 ectopic pregnancies requiring surgery after failed MTX treatment, 55 (59%) underwent planned salpingostomy. From 693 ectopic pregnancies managed without MTX, 166 were analyzed in detail, of which 80 (48%) underwent planned salpingostomy. The patients who underwent planned salpingostomy were thinner (body mass index 27.3 ± 7.2 kg/m2 vs 29.3 ± 8.3 kg/m2; p = .048), less frequently African American (33% vs 47%; p = .017), and more likely to have a visualized adnexal lesion (70% vs 52%; p = .004) than those undergoing planned salpingectomy. Preoperative ultrasound identified fetal cardiac activity and hemoperitoneum at comparable rates. MTX exposure was not associated with age, body mass index, race, ectopic risk factors, human chorionic gonadotropin levels, or gestational age at diagnosis, but the patients treated with MTX underwent surgery later than those who were untreated (gestational age 53.4 ± 11.2 days vs 43.5 ± 11 days; p <.001). The differences between the adnexal lesion size and rates of fetal cardiac activity and hemoperitoneum on ultrasound related to MTX exposure did not meet significance. Planned salpingostomy was completed in 22 (40%) of the patients treated with MTX vs 34 (42%) of those who were untreated. The reasons for failure, surgery time, and rates of hemoperitoneum or ectopic rupture were not associated with MTX exposure. Body mass index, race, tubal anastomosis history, visualization of the adnexal lesion, and MTX exposure were not significantly associated with the salpingostomy rate in a multivariate logistic regression model, but having a subspecialist surgeon (odds ratio 2.70; 95% confidence interval, 1.08-6.76; p = .033) and tubal rupture at surgery (odds ratio 0.23; 95% confidence interval, 0.09-0.54; p = .001) were. The initial medical management of an ectopic pregnancy with MTX is not associated with a decreased salpingostomy success rate.


Methadone salts. Part 1: preparation and identification.
The preparation of two methadone salts, namely: methadone-alpha-naphthalenesulfonate and methadone-o-benzoyl-benzoate is described. The prepared compounds were confirmed by t.l.c. and IR spectroscopic studies.


Etiological study of superficial radial nerve neuropathy: series of 34 patients.
Superficial radial nerve (SRN) neuropathy is a rare focal neuropathy leading to pain and paresthesia of the dorsolateral aspect of the hand. Reported causes include trauma, extrinsic compression, or it may be idiopathic. We describe the clinical and electrodiagnostic (EDX) features of 34 patients with SRN neuropathy of varied etiology. This is a retrospective study of patients with upper limb neuropathy referred for EDX studies who were found to have SRN neuropathy based on clinical and EDX findings. Twelve patients also had ultrasound (US) evaluations. Decreased pinprick sensation was noted in the distribution of the SRN in 31 (91%) patients, and a positive Tinel's sign was observed in 9 (26%). Sensory nerve action potentials (SNAPs) were not recordable in 11 (32%) patients. Of the patients who had a recordable SNAP, the latency was delayed, and the amplitude was decreased in all cases. Of the 12 patients who underwent US studies, 6 (50%) had an increased cross-sectional area of the SRN at or immediately proximal to the site of injury/compression. A cyst was located adjacent to the SRN in 2 patients. The most common cause of SRN neuropathy was trauma in 19 (56%) patients, of which 15 were iatrogenic. A compressive etiology was identified in 6 patients (18%). No specific etiology was detected in 10 patients (29%). This study is aimed at raising the awareness among surgeons about the clinical features and varied causes of SRN neuropathy; such knowledge may potentially lessen iatrogenic causes of injury.


Phosphorescent ruthenium complexes with bromopyrene unit that enhance oxygen sensitivity.
Ruthenium complexes are very useful phosphorescent probes for the visualization of hypoxia. We designed and synthesized three ruthenium complexes possessing bromopyrene, naphthalene, or anthracene units to improve the oxygen response. These ruthenium complexes provided strong phosphorescence under hypoxic conditions, while an increase in oxygen concentration led to a decrease in phosphorescence intensity. Among the ruthenium complexes, that with a bromopyrene unit (Ru-BrPy) had the best properties. This showed good cellular uptake and bright emission in cells, and had the highest sensitivity for molecular oxygen. Thus, Ru-BrPy is a promising candidate as a molecular probe for detecting cellular hypoxia.


Fibrous Dysplasia With Aneurysmal Bone Cyst Presenting as Sinonasal Mass.
Craniofacial involvement in fibrous dysplasia is a rare occurrence and complications of it are even rarer. Involvement of the sinonasal region can pose a challenge in both diagnosis and management. We hereby present a case of craniofacial fibrous dysplasia complicated with secondary aneurysmal bone cyst formation in a 15-year-old male patient.


Development of the thalamic reticular and perireticular nuclei in rats and their relationship to the course of growing corticofugal and corticopetal axons.
This study examines the connections of the thalamic reticular and perireticular nuclei during development. In addition, because these nuclei lie directly in the path of corticofugal and corticopetal axons during development, we have examined the relationship of these growing axons to the reticular and perireticular cell groups. Neurones were labelled by applying DiI, wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP), or HRP to the dorsal thalamus and/or cerebral cortex of rats at different stages of development. The axons of neurons in the reticular nucleus reach the dorsal thalamus as early as embryonic day (E) 14. At this age, and during later prenatal development, a small DiI implant limited to the presumptive lateral geniculate nucleus labels reticulothalamic and thalamocortical axons which travel in a clearly defined bundle through the thalamus. During late gestation, thalamocortical (approximately E15) and corticothalamic (approximately E17) axons pass directly through the reticular nucleus toward their targets. It is not until birth that collaterals are seen extending into the nucleus from the parent axons. Neurones in the perireticular nucleus, in contrast to those in the reticular nucleus, are not labelled from the lateral geniculate nucleus until after birth. The perireticular nucleus is very large at a stage when the first thalamocortical axons leave and when the first corticothalamic axons approach the thalamus. These axons are seen to change course sharply in the region of the internal capsule, where there are many perireticular cells. Corticothalamic axons turn toward the reticular nucleus, and thalamocortical axons turn toward the cortical subplate. Corticospinal and corticobulbar axons, on the other hand, pass directly through the perireticular region toward their more caudal targets. After these axons have reached their targets, the perireticular nucleus reduces dramatically in size.


Paratesticular leiomyoma with a der(14)t(12;14)(q15;q24).
While uterine leiomyomas are among the most common and best cytogenetically characterized solid tumors, leiomyomas at other sites are rare. Only two karyotypically abnormal leiomyomas in males have been reported to date, both of them with unspecific chromosome aberrations. We recently analyzed by G-banding a paratesticular leiomyoma, a tumor type not cytogenetically examined before, and found the pseudodiploid karyotype 46,XY,der(5)t(5;14)(q31;q24),der(14)t(12;14)(q15;q24). The leiomyoma cells demonstrated strong immunohistochemical nuclear expression of the HMGA2 protein, supporting a role of HMGA2 as the target gene in 12q14∼15 rearrangements. In uterine leiomyomas, the t(12;14)(q15;q24) is the most frequent translocation leading to overexpression of HMGA2, therefore it seems that a common pathogenetic pathway exists for benign smooth muscle tumors of both the female and male reproductive organs. The finding of this abnormality may help identify a scrotal tumor of uncertain biologic potential but with smooth muscle differentiation as benign.


Effects of carbonyl cyanide p-trichloromethoxyphenylhydrazone (CCCP) and of ruthenium red (RR) on capsaicin-evoked neuropeptide release from peripheral terminals of primary afferent neurones.
In the superfused isolated rat urinary bladder, capsaicin as well as electrical field stimulation evoked the release of calcitonin gene-related peptide-like immunoreactivity (CGRP-IR). Carbonyl cyanide p-trichloromethoxyphenylhydrazone (CCCP, threshold 2 microM) reduced both, the capsaicin- and the electrical field stimulation-evoked release of CGRP-IR while a low concentration of Ruthenium Red (RR, 0.6 microM and 2 microM) selectively attenuated the capsaicin-evoked release of CGRP-IR but did not influence the effect of electrical field stimulation. 20 microM RR nearly abolished the capsaicin-evoked release, but also attenuated the effect of electrical field stimulation. In the isolated guinea-pig bronchus, electrical field stimulation and capsaicin induced non-cholinergic contractions which are known to be caused by tachykinin release from afferent nerve terminals. CCCP (0.6 microM) only reduced the response to field stimulation; a ten-fold higher concentration of CCCP attenuated field stimulation as well as capsaicin-induced contractions. This is in contrast to the reported selective inhibition of capsaicin-induced contractions by RR. The present data demonstrate that CCCP generally inhibits evoked neuropeptide release, regardless of the kind of stimulation used while low concentrations of RR preferentially inhibit capsaicin-evoked neuropeptide release.


Dendritic integration: 60 years of progress.
Understanding how individual neurons integrate the thousands of synaptic inputs they receive is critical to understanding how the brain works. Modeling studies in silico and experimental work in vitro, dating back more than half a century, have revealed that neurons can perform a variety of different passive and active forms of synaptic integration on their inputs. But how are synaptic inputs integrated in the intact brain? With the development of new techniques, this question has recently received substantial attention, with new findings suggesting that many of the forms of synaptic integration observed in vitro also occur in vivo, including in awake animals. Here we review six decades of progress, which collectively highlights the complex ways that single neurons integrate their inputs, emphasizing the critical role of dendrites in information processing in the brain.