Neutrophil exosomes enhance the skin autoinflammation in generalized pustular psoriasis via activating keratinocytes.
Generalized pustular psoriasis (GPP) is a rare and severe inflammatory skin disease that can be life-threatening. Gene mutations are found in some cases, but its immune pathogenesis is largely unknown. Here, we observed that the neutrophil:lymphocyte ratio in patients with GPP was higher than that in healthy controls and decreased after effective treatment. Neutrophils isolated from patients with GPP induced higher expressions of inflammatory genes including IL-1β, IL-36G, IL-18, TNF-α, and C-X-C motif chemokine ligands in keratinocytes than normal neutrophils did. Moreover, neutrophils from patients with GPP secreted more exosomes than controls, which were then rapidly internalized by keratinocytes, increasing the expression of these inflammatory molecules via activating NF-κB and MAPK signaling pathways. The proteomic profiles in neutrophil exosomes further characterized functional proteins and identified olfactomedin 4 as the critical differentially expressed protein that mediates the autoimmune inflammatory responses of GPP. These results demonstrate that neutrophil exosomes have an immune-regulatory effect on keratinocytes, which modulates immune cell migration and autoinflammation in GPP.-Shao, S., Fang, H., Zhang, J., Jiang, M., Xue, K., Ma, J., Zhang, J., Lei, J., Zhang, Y., Li, B., Yuan, X., Dang, E., Wang, G. Neutrophil exosomes enhance the skin autoinflammation in generalized pustular psoriasis via activating keratinocytes.


Visual evoked potentials in mild senile dementia of Alzheimer type.
Visual evoked potentials (VEPs) to chessboard shift and to flash stimulation were recorded from 40 subjects with senile dementia of Alzheimer type (SDAT) and 40 individually matched control subjects. All of the SDAT subjects had only a mild degree of dementia and were still living in the community. Analysis of variance showed significant differences between demented and control group means for 3 chessboard shift VEP measures, the demented group having longer latency of peaks P3 and N3, and larger amplitude of segment N2-P3. These three are the earliest reported changes in the VEP in Alzheimer disease, since the flash VEPs showed no measure in which the demented and control group means differed significantly.


A single nucleotide polymorphism in the MMP-1 promoter is correlated with histological differentiation of gastric cancer.
Matrix metalloproteinase-1 (MMP-1) plays a key role in cancer invasion and metastasis by degradation of extracellular matrix (ECM) and basement membrane barriers. The 1G/2G single nucleotide polymorphism (SNP) in the MMP-1 promoter at position -1607 bp has been reported to affect the transcriptional activity. In the light of these findings, we investigated whether this SNP in the MMP-1 promoter is associated with the development, differentiation, and progression of gastric cancer. The 215 gastric cancer patients and 166 controls were used in this study. The SNP of the MMP-1 promoter was analyzed by PCR-RFLP and sequencing. The genotype frequency was compared between cases and controls, and the association with clinicopathological parameters among cases was studied. The frequency of 1G/2G genotypes in gastric cancer patients was similar to those in controls (p=0.57). The degree of tumor invasion, the presence of lymph node metastasis, and clinical stage showed no significant association with the SNP. On the other hand, we found a significant association with histological differentiation and gender among gastric cancer patients (p<0.05, respectively). The presence of 2G allele in the MMP-1 promoter did not enhance the risk of gastric cancer; however, it may be involved in differentiation of gastric cancer.


Structural Basis for Virulence Activation of Francisella tularensis.
The bacterium Francisella tularensis (Ft) is one of the most infectious agents known. Ft virulence is controlled by a unique combination of transcription regulators: the MglA-SspA heterodimer, PigR, and the stress signal, ppGpp. MglA-SspA assembles with the σ70-associated RNAP holoenzyme (RNAPσ70), forming a virulence-specialized polymerase. These factors activate Francisella pathogenicity island (FPI) gene expression, which is required for virulence, but the mechanism is unknown. Here we report FtRNAPσ70-promoter-DNA, FtRNAPσ70-(MglA-SspA)-promoter DNA, and FtRNAPσ70-(MglA-SspA)-ppGpp-PigR-promoter DNA cryo-EM structures. Structural and genetic analyses show MglA-SspA facilitates σ70 binding to DNA to regulate virulence and virulence-enhancing genes. Our Escherichia coli RNAPσ70-homodimeric EcSspA structure suggests this is a general SspA-transcription regulation mechanism. Strikingly, our FtRNAPσ70-(MglA-SspA)-ppGpp-PigR-DNA structure reveals ppGpp binding to MglA-SspA tethers PigR to promoters. PigR in turn recruits FtRNAP αCTDs to DNA UP elements. Thus, these studies unveil a unique mechanism for Ft pathogenesis involving a virulence-specialized RNAP that employs two (MglA-SspA)-based strategies to activate virulence genes.


Impact of early risk stratification on the length of hospitalization in patients with acute Q-wave myocardial infarction. 'The 60-minutes myocardial infarction project'.
An assessment of individual risk factors may identify a subgroup of postinfarction patients at low risk, i.e. patients appropriate for early discharge. Using a large unselected population of the national registry, 'The 60-Minutes Myocardial Infarction Project', we (1) attempted to provide a retrospective analysis of clinical factors and in-hospital mortality in a population living on the 6th hospital day following admission to define a low-risk patient group with a residual in-hospital mortality of less than 1% eligible for early discharge, and (2) to analyze the current impact of risk stratification based on these clinical factors on the length of hospitalization. The study group consisted of 12,045 survivors on the 6th day after admission out of 14,980 patients of the registry with proven Q-wave myocardial infarction. Risk modeling was performed with multiple logistic regression. A total of 873 patients (7.3%) died after day 6 in hospital. The most important prognostic factors were cardiopulmonary resuscitation prior to admission (odds ratio, OR: 7.2, confidence interval, CI: 5.11-10.22), thrombolysis complicated by severe bleedings (OR: 6.2, CI: 1.2-31. 2) and age >70 years (OR 4.7, CI 3.51-6.39). The other more significant independent predictors of increased mortality were end-stage renal disease, age between 56 and 70 years, systolic blood pressure <95 mm Hg on admission, history of trauma </=2 months, cancer and left-bundle-branch block. Summarizing these nine groups of patients with the strongest association to in-hospital mortality, we defined a high-risk group comprising 79% of the AMI patients with a residual in-hospital mortality of 8.8%. On the other hand, by excluding these nine high-risk patient groups, a low-risk group of 21% of all AMI patients seems to be appropriate for early discharge (residual in-hospital mortality = 1.07%). However, in the current practice, there was no difference regarding the median length of hospital stay between the two risk groups. The low-risk patients were hospitalized 20 days compared to 22 days in the high-risk patients. Using a simple logistic regression model, which considers clinical factors of the early hospital phase, one fifth of the infarction patients can be stratified to be at low risk, and might be eligible for early hospital discharge. Currently, an individual risk stratification has no impact on the length of hospital stay in Germany.


Regulation of PLCbeta1a membrane anchoring by its substrate phosphatidylinositol (4,5)-bisphosphate.
Basic knowledge as to the subcellular location and dynamics of PLCbeta isozymes is scant. Here, we report on the subcellular location of GFP-PLCbeta1a and the use of total internal reflection fluorescence (TIRF) microscopy to examine the dynamics of GFP-PLCbeta1a at the plasma membrane upon stimulation of Gq-coupled receptors. Using this technique, we observed PLCbeta1a dissociation from the plasma membrane upon addition of agonist. An increase in intracellular calcium and a decrease in PtdIns(4,5)P2 both coincided with a translocation of PLCbeta1a from the plasma membrane into the cytosol. In order to differentiate between calcium and PtdIns(4,5)P2, rapamycin-induced heterodimerization of FRB and FKBP12 fused to 5-phosphatase IV was used to instantaneously convert PtdIns(4,5)P2 into PtdIns(4)P. Addition of rapamycin caused PLCbeta1a to dissociate from the plasma membrane, indicating that removal of PtdIns(4,5)P2 is sufficient to cause translocation of PLCbeta1a from the plasma membrane. In conclusion, PLCbeta1a localization is regulated by its own substrate.


Relevance of Intraparotid Metastases in Head and Neck Skin Squamous Cell Carcinoma.
Parotid lymph node metastases are common in head and neck cutaneous squamous cell carcinoma (cSCCHN). Here we evaluate the diagnostic, prognostic, and therapeutic implications of intraglandular lymph node (IGLN) metastases in cSCCHN. Retrospective study in a tertiary referral university cancer institute. We included patients with cSCCHN who underwent parotidectomy and neck dissection (ND), with or without synchronous resection of the skin primary, between January 1999 and January 2018. The characteristics of cSCCHN with or without IGLN involvement were compared. Altogether, 68 patients were included. Of the 29 (42.6%) patients classified as cN0, eight were upstaged pN+ and had concomitant IGLN involvement. Of 21 patients with pN0 disease, IGLN metastases were absent in only three cases, resulting in a specificity and sensitivity of parotid metastases to diagnose occult nodal neck metastases of 14.29% and 100%, respectively. The positive and negative predictive values were 14.29% and 100%, respectively. Univariate analyses only displayed a significantly higher rate of moderately and poorly differentiated primaries in patients with IGLN metastases (P = .015). Only advanced T-stages were significantly associated with neck recurrences. IGLN status in advanced cSCCHN is potentially predictive for occul nodal neck metastases. Our results suggest that ND in patients with histopathologically negative IGLNs and clinically negative neck lymph nodes may not be necessary given the high negative predictive value of IGLN status in this group of patients. Therefore, accurate diagnostic evaluation of IGLN involvement is mandatory. 4 Laryngoscope, 131:788-793, 2021.


Profiling kidney microRNAs from juvenile grass carp (Ctenopharyngodon idella) after 56days of oral exposure to decabromodiphenyl ethane.
Grass carp (Ctenopharyngodon idella) is one of the most important species in China. Decabromodiphenyl ethane (DBDPE) is a brominated flame retardant that has been used widely in industry, and has been observed to accumulate in the tissues of fish from South China. Evidence has shown that DBDPE is toxic to aquatic animals, but the molecular response has been unclear. MicroRNAs (miRNAs) are small noncoding and negative regulatory RNAs that are 20-24 nucleotides in length, which are involved in a wide range of biological processes. We took advantage of deep-sequencing techniques to accurately and comprehensively profile the kidney miRNA expression of grass carp after 8weeks of oral exposure to DBDPE. After mapping sequencing data to the genome and Expressed Sequence Tags (ESTs) of grass carp, we identified 493 miRNAs in the sequenced grass carp samples, which included 51 new miRNAs. The results indicated that 5 miRNAs were significantly down-regulated and 36 miRNAs were significantly up-regulated (FDR<0.001, 1.5-fold change) after DBDPE exposure. Real-time quantitative PCR (RT-qPCR) was performed on 4 miRNAs from the two samples, and the sequencing and RT-qPCR data were consistent. This study provides the first comprehensive identification of grass carp miRNAs, and the first expression analysis of grass carp miRNAs following DBDPE exposure. The results indicated that miRNAs have potential for use as biomarkers.


Biogenesis of constitutive secretory vesicles, secretory granules and synaptic vesicles.
The formation of constitutive secretory vesicles and secretory granules from the trans-Golgi network is inhibited by brefeldin A, suggesting a role for ADP ribosylation factor and other coat proteins in the budding process, and is regulated by multiple heterotrimeric G proteins. Membrane proteins are sorted to secretory granules through signals in their cytoplasmic domains and, like secretory proteins, by aggregation of their lumenal domains. Synaptic vesicle membrane proteins undergo exocytotic-endocytotic recycling before synapse formation, consistent with the proposed biogenesis of synaptic vesicles from early endosomes.


Omission of non-poly(A) viral transcripts from the tissue level atlas of the healthy human virome.
A recent paper in BMC Biology entitled "A tissue level atlas of the healthy human virome" by Kumata et al. describes a meta-transcriptomic analysis of RNA-sequencing datasets from the Genotype-Tissue Expression (GTEx) Project. Using a workflow that maps the GTEx sequences to the human genome, then screens unmapped sequences to detect viral transcripts, the authors present a quantitative analysis of the presence of different viruses in the non-diseased tissues of over 500 individuals and assess the impact of these viruses on host gene expression. Here we draw attention to an issue not acknowledged in this study. Namely, by relying solely on GTEx datasets, which are enriched for transcripts with poly(A) tails, the analysis will have missed non-poly(A) viral transcripts, rendering this tissue level atlas of the virome incomplete.A commentary on Kumata et al. (BMC Biol 18:55, 2020).


Similar long-term recurrence rates with cystectomy and CO2 laser vaporization for endometrioma: a retrospective study.
Is there a difference in recurrence rate of endometrioma(s) after cystectomy versus CO2 laser vaporization of the cyst wall? This single-centre retrospective study included 270 patients undergoing laparoscopic surgery for endometriomas between January 2010 and December 2014, stratified according to the surgical technique used. All 270 included patients underwent complete laparoscopic surgery for endometrioma(s): 155 underwent cystectomy, 63 complete CO2 laser vaporization of the cyst wall and 52 a mixed technique. The primary outcome studied was the difference in recurrence rate between the cystectomy group and the CO2 laser vaporization group. The mean duration of follow-up was 58 (±34) months. Imaging-based recurrence (any cyst size) was reported in 9.9% of patients (n = 12/121) treated with cystectomy and in 13.3% of patients (n = 6/45) who underwent a vaporization (P = 0.577). The need for reintervention for endometrioma(s) was also similar in both groups, with a rate of 3.2% (n = 5/155) after cystectomy and 4.8% (n = 3/63) after vaporization (P = 0.476). Of 160 women who wanted to conceive immediately after surgery, 73.8% became pregnant (72.6% [77/106] in the cystectomy group and 75.9% [41/54] in the vaporization group [P = 0.310]). Conception occurred mostly by assisted reproductive technology (57.1% [44/77] in the cystectomy group and 70.7% [29/41] in the vaporization group [P = 0.074]). Similar rates of recurrence for endometrioma(s) were observed after cystectomy versus CO2 laser vaporization. As other studies have suggested that CO2 laser vaporization may be less harmful to the normal ovarian tissue, it can be considered as a safe alternative for cystectomy in women wishing to preserve their reproductive potential.


Sarcomatoid carcinoma after radiation treatment of prostatic adenocarcinoma.
We report 2 patients with conventional prostatic adenocarcinoma who developed sarcomatoid carcinoma of probable prostatic origin 6 and 2.5 years after radiation treatment (seed implantation and external beam). Our cases had histologic features consistent with those cases previously reported in the literature. The tumors consisted of spindle cells with large hyperchromatic nuclei and a pattern mimicking a sarcoma. Immunohistochemical studies showed the tumors to be weakly positive for EMA, CK7, and vimentin. Ki67 staining showed positivity in more than 50% of tumor cells. The tumors also stained diffusely positive for p53 and p63. PSA and PAP were negative. Clinically, the sarcomatoid carcinomas appeared to be of prostatic origin. The pathogenesis of the tumors is still uncertain but most likely represent a radiation-induced dedifferentiation of prostatic adenocarcinoma.


Sulphite stabilization and high-performance liquid chromatographic determination: a reference method for free and reversibly bound sulphite in food.
A high-performance liquid chromatographic (HPLC) post-column reactor combination has been developed for the determination of sulphite (free and reversibly bound) in 15 different foods. The foods were treated with a pH 5.1 buffered aqueous 2% formaldehyde solution to convert the labile sulphite to the relatively stable hydroxymethylsulphonate. Reverse-phase ion-pairing HPLC with a post-column detector consisting of an initial reaction with KOH followed by colorimetric reaction with Ellman's reagent buffered at pH 6.3 were used for separation and quantitation. The photometric detector at 412 nm quantitated the strongly absorbing 3-carboxy-4-nitrothiophenolate anion, the product of the sulphite reaction with Ellman's reagent. The recovery at levels of 5-100 ppm as sulphur dioxide was greater than 90% by reverse isotope dilution assay. Repetitive analysis of a single grape juice extract containing 20 ppm SO2 showed a relative standard deviation of 2.2%.


Unrecognized aspiration of an oropharyngeal airway.
A case of unsuspected aspiration of an oropharyngeal airway causing dysphagia is described. Visualization of this airway on plain film would be facilitated if radio-opaque material were included at manufacture.


[Surgery for carcinoma of the oesophagus. A plea for precise technique and pleuro-mediastinal drainage (author's transl)].
During surgery for carcinoma of the oesophagus, the authors emphasise the need for selective intubation using a Carlens tube, of avoiding damage to the diaphragm and the phrenic nerve at all levels, and of draining the hemithorax involved in the operation by three drains: anterior and posterior thoracic and madiastino-abdominal. These precautions reduce post-operative difficulties, justifiy excision surgery for carcinoma of the oesophagus, and to a certain extent make it possible to reduce the contraindications, in particular in the presence of tracheo-bronchial spread. Amongst 107 patients undergoing surgery, in whom there was a risk of respiratory insufficiency in one quarter, surgery (sometimes with extension of the excision: one case in four) was associated with an operative and post-operative mortality of 18%.


Nodular regenerative hyperplasia of the liver. Report of six cases and review of the literature.
The authors report the cases of 6 adult patients suffering from nodular regenerative hyperplasia of the liver, an uncommon lesion characterized by small-sized hepatocytic nodules scattered throughout the liver, with no or slight fibrosis. The histological recognition of nodular regenerative hyperplasia of the liver was easy on large specimens taken intraoperatively, but was difficult or impossible on small specimens such as those provided by needle biopsy. In all of the patients of this series, the main consequence of nodular regenerative hyperplasia of the liver was portal hypertension. In this series, nodular regenerative hyperplasia of the liver was not associated with rheumatoid arthritis, whereas such an association has been noted in most of the cases previously reported. The etiology and pathogenesis of nodular regenerative hyperplasia of the liver is unknown.


Three-dimensional vectorcardiography (3-D VCG) by computer graphics in old myocardial infarction.
By using computer graphics we rotated the vector loop and three coordinate axes to find the viewpoint where the infarctional changes are maximally exposed and demonstrated the advantage of the "3-D VCG" over the conventional VCG by defining the quantitative "MI index." The orthogonal electrocardiogram recorded by the Frank lead system was digitally measured and processed by a microcomputer. The loops and axes were rotated about the X axis (X-rot) and the Y axis (Y-rot). The spatial vector loop and orthogonal coordinates can be presented as viewed from any spheric direction. Eight quadrants were illustrated with four colors and red circles. The subjects consisted of 30 patients with old anterior myocardial infarction (MI) and 15 patients with old inferior MI. We measured the area of "Bite" in anterior MI and superior displacement in inferior MI. The MI index was defined and averaged in 361 directions. In anterior MI, the maximum mean index was obtained when X-rot is +90 degrees and Y-rot -40 degrees, viewed from upward and leftward, whereas in inferior MI it was obtained when X-rot is -50 degrees and Y-rot -80 degrees, viewed from downward and leftward. These values were significantly higher than those in conventional VCG projections, substantiating superior diagnostic sensitivity of 3-D VCG.


Congenital Bronchial Stenosis Presenting as Neonatal Respiratory Distress: A Case Report.
Neonatal respiratory distress has a broad differential that includes cardiac, pulmonary, anatomic, and infectious etiologies. Congenital stenotic lesions of the trachea and bronchus are rare and can occur anywhere along the tracheobronchial tree. Patients with tracheobronchial stenosis typically present in the neonatal period with respiratory distress. We present a case of a 10-day-old term female who presented to the emergency department (ED) with tachypnea and increased work of breathing. She was found to have congenital bronchial stenosis of her right mainstem bronchus. She was stabilized in the ED and remained in the neonatal intensive care unit until successful slide tracheoplasty was performed. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: Congenital bronchial stenosis is a rare etiology of respiratory distress in a neonate. Anatomic lower airway abnormalities are an important cause of neonatal tachypnea and must remain on the differential. In addition to respiratory stabilization with noninvasive or invasive support, evaluation should be directed at determining the location and anatomic characteristics of the area of stenosis.


Association between interleukin 8 -251 A/T and +781 C/T polymorphisms and osteosarcoma risk in Chinese population: a case-control study.
Interleukin-8 (IL-8) is an angiogenic chemokine that plays a potent role in both development and progression of many human malignancies. However, there are no data about the role of IL-8 polymorphism in development of osteosarcoma. A hospital-based case-control study was conducted among 190 patients with osteosarcoma and 190 healthy controls to investigate the possible association between the IL-8 -251 A/T and +781 C/T polymorphisms, respectively, and the risk of osteosarcoma. Significant differences of genotype distribution were observed between osteosarcoma cases and controls at the IL-8 -251T/A genotypes. Compared with the IL-8 -251T/A homozygote TT, the heterozygous TA genotype was associated with significantly increased risk for osteosarcoma (odds ratio (OR) = 2.16, 95 % confidence interval (CI) = (1.38-4.52), P = 0.021); the AA genotype was associated with increased risk for osteosarcoma (OR = 1.94, 95 % CI = 1.31-3.83, P = 0.018). TA and AA combined variants were associated with increased risk for osteosarcoma compared with the TT genotype (OR = 1.72, 95 % CI = 1.45-4.41, P = 0.023). Moreover, the genotype AA of IL-8 -251T/A carried a higher risk of osteosarcoma metastasis and later Enneking stages, compared with the TT genotype. However, the genotype and allele frequencies of IL-8 +781 C/T polymorphisms in osteosarcoma patients were not significantly different from controls. Our results showed that the IL-8 -251 A/T genotype was associated with increased risk for development and metastasis of osteosarcoma in Chinese Han population.


On the predictability of short-lived particulate matter around a cement plant in Kerman, Iran: machine learning analysis.
One of the greatest environmental risks in the cement industry is particulate matter emission (i.e., PM2.5 and PM10). This paper aims to develop descriptive-analytical solutions for increasing the accuracy of predicting particulate matter emissions using resample data of Kerman cement plant. Photometer instruments DUST TRAK and BS-EN-12341 method were used to determine concentration of PM2.5 and PM10. Sampling was performed on 4 environmental stations of Kerman cement plant in the four seasons. In order to accurate assessment of particulate matter concentration, a new model was proposed to resample cement plant time series data using Pandas in Python. The effect of meteorological parameters including wind speed, relative humidity, air temperature and rainfall on the particulate matter concentration was investigated through statistical analysis. The results indicated that the maximum annual average of 24-h of PM2.5 belonged to the east side (opposite the clinker depot) in 2019 (31.50 μg m-3) and west side (in front of the mine) in 2020 (31.00 μg m-3). Also, maximum annual average of 24-h of PM10 belonged to the west side (in front of the mine) in 2020 (121.00 μg m-3) and east side (opposite the clinker depot) in 2020 (120.75 μg m-3). The PM2.5 and PM10 concentrations are more than the allowable limit. The results demonstrate that particulate matter concentration increases with increasing relative humidity and rainfall. Finally, the SARIMA model was used to predict the particulate matter concentration. The online version contains supplementary material available at 10.1007/s13762-022-04645-3.


Effects of age simulation and age on motor sequence learning: Interaction of age-related cognitive and motor decline.
Aging is known to lead to decrements in sensory and cognitive functioning and motor performance. The purpose of the present experiment was twofold: a) We assessed the influence of wearing an age simulation suit on motor sequence learning, cognitive speed tasks and far visual acuity in healthy, younger adults. b) We evaluated the interaction of cognitive aging and declining motor sequence learning in older adults. In a between-subjects design we tested 11 younger adults (Mage = 23.6 years) without the age suit, 12 younger adults wearing the age suit (Mage = 23.2 years), and 23 older adults (Mage = 72.6 years). All participants learned a simple, spatial-temporal movement sequence on two consecutive days, and we assessed perceptual processing speed (Digit Symbol Substitution test and Figural Speed test) and far visual acuity. Wearing an age simulation suit neither affected the learning of the simple motor sequence nor the performance at the cognitive speed tasks in younger adults. However, far visual acuity suffered from wearing the suit. Younger adults with and without the suit showed better motor sequence learning compared to older adults. The significant correlations between the cognitive speed tests and the motor learning performance in older adults indicated that cognitive aging partially explains some of the variance in age-related motor learning deficits.


A Brief Review of Paradigm Shifts in Prevention of Alzheimer's Disease: From Cognitive Reserve to Precision Medicine.
Alzheimer's disease (AD) and related dementias can be an enormous economic burden for taxpayers, patients, their families, medical systems, and society as a whole. Since disease-modifying treatments have failed, several studies have instead focused on a paradigm shift for preventing and treating AD. A higher cognitive reserve (e.g., greater education, occupational attainment, or more leisure activities) is associated with protection against disease-related cognitive decline. Precision medicine aims to optimize the effectiveness of disease prevention and treatment by considering specific biological components of individuals. We suggest that research into cognitive reserve and precision medicine could be a key to overcoming the limitations of traditional approaches to the prevention and treatment of AD.


Detection of cytokeratin 20 mRNA in the peripheral blood of patients with colorectal cancer by immunomagnetic bead enrichment and real-time reverse transcriptase-polymeras chain reaction.
Detection of circulating cancer cells is a useful indicator for the risk of recurrence of advanced carcinoma. The aim of the present study was to evaluate the potential value of a novel approach to detect the circulating cancer cells in patients with colorectal cancer. This method is based on a combination of isolation of epithelial cell by a combination of negative and positive immunomagnetic beads with detection of cytokeratin 20 (CK20) mRNA by reverse transcriptase-polymeras chain reaction (RT-PCR). Peripheral blood samples were collected from 40 patients with colorectal carcinoma on the day before operation or chemical therapy. Mononuclear cells (MNC) were isolated by centrifugation through a Ficoll gradient. Each MNC sample was equally divided into three parts and then CD45 immunomagnetic beads and/or Ber-EP4 immunomagnetic beads were used to enrich colon cancer cells. Finally, the CK20 mRNA was detected by real-time quantitative RT-PCR. As a control, LS174T colon cancer cells were serially diluted with blood from healthy individuals. When CD45 and Ber-EP4 immunomagnetic beads were used successively, a significant correlation between CK20 mRNA levels and the initial cell concentrations was found in the control recovery experiment. The sensitivity of the assay was one cancer cell in 1 mL healthy blood. In the patient group, CK20 mRNA was detected in 80.0%, 82.5% and 72.5% of patients when CD45, Ber-EP4, and CD45/Ber-EP4 immunomagnetic beads were used, respectively. The positive detection rates of patients with colorectal carcinoma at Dukes A, B, C, and D stage were 0.0% (0/2), 33.3% (3/9), 86.7% (13/15), and 92.9% (13/14), respectively. The CK20 mRNA positive detection rate in peripheral blood was significantly correlated with tumor diameter (P < 0.01, chi(2)), lymphatic metastasis (P < 0.05) and hepatic metastasis (P < 0.05), but not with the differentiation of tumor cells. The combined use of negative and positive immunomagnetic beads followed by amplification of CK20 mRNA by means of RT-PCR is a non-invasive, sensitive, and specific assay for the detection of circulating colonic cancer cells.


Impact of arsenic trioxide in the treatment of acute promyelocytic leukemia.
Arsenic trioxide (ATO) is presently the most active single agent in the treatment of acute promyelocytic leukemia (APL). This review provides insights into the mode of action and the pharmacological properties of ATO, and summarizes the most relevant results of more than 20 treatment studies in relapsed or newly diagnosed APL published between 1997 and 2011. ATO acts by targeting multiple pathways in APL leading to apoptosis and myeloid differentiation. It induces complete remission without myelosuppression and causes only few adverse effects. In relapsed APL, ATO-based salvage therapy has been able to induce long-lasting remissions and possible cure in 50-81% of patients. In newly diagnosed APL, two main strategies are currently pursued. ATO is either included into induction therapy with the aim to minimize or eliminate chemotherapy, or it is incorporated as an additive into established first-line concepts with all-trans-retinoic acid and chemotherapy to reinforce their anti-leukemic efficacy. Recent results suggest a high efficacy of ATO in both concepts. In conclusion, experimental research and clinical studies have made contributions toward a better understanding of the molecular mechanisms induced by ATO in APL cells and have established this historic substance as an important candidate for the further improvement of APL therapy.


The association between event learning and continuous quality improvement programs and culture of patient safety.
To present our approach and results from our quality and safety program and to report their possible impact on our culture of patient safety. We created an event learning system (termed a "good catch" program) and encouraged staff to report any quality or safety concerns in real time. Events were analyzed to assess the utility of safety barriers. A formal continuous quality improvement program was created to address these reported events and make improvements. Data on perceptions of the culture of patient safety were collected using the Agency for Health Care Research and Quality survey administered before, during, and after the initiatives. Of 560 good catches reported, 367 could be ascribed to a specific step on our process map. The calculated utility of safety barriers was highest for those embedded into the pretreatment quality assurance checks performed by physicists and dosimetrists (utility score 0.53; 93 of 174) and routine checks done by therapists on the initial day of therapy. Therapists and physicists reported the highest number of good catches (24% each). Sixty-four percent of events were caused by performance issues (eg, not following standardized processes, including suboptimal communications). Of 31 initiated formal improvement events, 26 were successfully implemented and sustained, 4 were discontinued, and 1 was not implemented. Most of the continuous quality improvement program was conducted by nurses (14) and therapists (7). Percentages of positive responses in the patient safety culture survey appear to have increased on all dimensions (p < .05). Results suggest that event learning and continuous quality improvement programs can be successfully implemented and that there are contemporaneous improvements in the culture of safety.


Solvothermal synthesis and properties control of doped ZnO nanoparticles.
Indium and gallium doped ZnO nanoparticles have been prepared by a hydrothermal reaction in ethanol and methoxyethanol. A comprehensive study of the preparation process, including a thorough investigation by TG-FTIR and TG-MS of the thermal-purification procedure, is presented. Moreover, the effect of thermal conditions and dopant concentration on the structural and optical properties is discussed on the basis of XRD, TEM and UV-vis-NIR results. Reported data indicated that the use of methoxyethanol as a solvent allows an enhanced control of nanoparticle size and favours dopant incorporation into zinc oxide. Near infrared absorption of these materials can be strongly affected by increasing the doping level and upon treating nanoparticles under reducing atmosphere. Preliminary study indicated that this effect is greatly enhanced for gallium-doped zinc oxide.


Reactive arthritis due to Salmonella schwarzengrund in a patient with asymptomatic spondylitis.
Reactive arthritis following enteric infection with Salmonella schwarzengrund, a species not previously described in association with this condition, occurred in a 51-year-old man with bilateral sacro-iliitis.


Immunophenotypic Features of Myeloid Neoplasms Associated with Chromosome 7 Abnormalities.
Abnormalities involving chromosome 7 are one of the most frequent chromosomal aberrations in myeloid neoplasms including myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) and are associated with an adverse prognosis. Immunophenotyping by flow cytometry provides data that can assist in the diagnosis and classification of myeloid neoplasms. The immunophenotypic features of myeloid neoplasms with monosomy 7 or del(7q) have not been previously described in a comprehensive fashion. We retrospectively analyzed flow cytometric data of myeloid neoplasms with monosomy 7 or del(7q) and summarized associated immunophenotypic features. Myeloid neoplasms with monosomy 7 typically demonstrate multiple immunophenotypic abnormalities on myeloid blasts and maturing myelomonocytic cells. Increased CD14 expression on maturing granulocytic cells was characteristically seen in myeloid neoplasms with monosomy 7. This abnormality was significantly more frequent in myeloid neoplasms with monosomy 7 than in those with del(7q) (92.9% vs. 8.2%, P < 0.0001). The presence of increased CD14 expression on maturing granulocytic cells could be seen in cases with monosomy 7 in the setting of a normal blast percentage and when minimal (<3) other immunophenotypic abnormalities were seen on myeloid populations. Increased CD14 expression on maturing granulocytic cells can be helpful in identifying patients with myeloid neoplasms who have monosomy 7. © 2019 International Clinical Cytometry Society.


Degradation of Verapamil hydrochloride in water by gliding arc discharge.
This study investigated the influence of gliding arc plasma discharge on the degradation of Verapamil hydrochloride in water. The plasma discharge was characterized by means of optical emission spectroscopy. Spectra of various atomic and molecular species were observed. Aqueous solution of Verapamil hydrochloride was exposed to gliding arc discharge operated in continuous discharge at atmospheric pressure and room temperature. The identification of Verapamil, the degradation mechanisms of Verapamil and its transformation products were performed using liquid chromatography - mass spectrometry (HPLC-MS). Experimental results indicate that the atmospheric pressure gliding arc plasma treatment has noticeable effects on Verapamil with satisfactory degradation efficiency. Plausible mechanisms of the degradation were discussed.


Wnt/beta-catenin signaling: new (and old) players and new insights.
Wnt/beta-catenin signaling has central roles in embryogenesis and human diseases including cancer. A central scheme of the Wnt pathway is to stabilize the transcription coactivator beta-catenin by preventing its phosphorylation-dependent degradation. Significant progress has been made toward the understanding of this crucial regulatory pathway, including the protein complex that promotes beta-catenin phosphorylation-degradation, and the mechanism by which the extracellular Wnt ligand engages cell surface receptors to inhibit beta-catenin phosphorylation-degradation. Here we review some recent discoveries in these two areas, and highlight some crucial questions that remain to be resolved.


Imaging of Diffuse Lung Disease in the Intensive Care Unit Patient.
There is a wide variety of causes of diffuse lung disease in the intensive care unit patient, of which adult respiratory distress syndrome is the commonest clinical consideration. Plain radiography, computed tomography, and ultrasound can be used synergistically to evaluate patients with diffuse lung disease and respiratory impairment. Imaging is not limited to characterization of the cause of diffuse lung disease but also aids in monitoring its evolution and in ventilator setting management.


No effect of cigarette smoking on electroencephalographic nonlinearity.
The Gaussian surrogate-date procedure was applied to the measurement of the effect of cigarette smoking on the dimensional complexity of normal, resting EEG. Evidence of significant nonlinearity in the EEG was obtained, replicating previous results. However, unlike EEG dimensional complexity, EEG nonlinearity (difference between original and surrogate data) was not affected by smoking. This indicates that, under resting conditions, smoking/nicotine may have a modulating effect on input from the reticular activating system, with such input having a global, linearizing effect on cortical dynamics. Nonlinear dynamics resulting from intracortical processes appear not to be affected.


Application of NGS molecular classification in the diagnosis of endometrial carcinoma: A supplement to traditional pathological diagnosis.
This study aims to demonstrate the advantages of NGS molecular classification in EC diagnosis and to assess whether molecular classification could be performed on curettage specimens and its concordance with subsequent hysterectomy specimens. 80 patients with hysterectomy specimens and 35/80 patients with paired curettage specimens were stratified as POLE mut, MSI-H, TP53 wt, or TP53 abn group by NGS panel. Histotype, tumor grade, IHC results, and other pathological details were taken from original pathological reports. The correlation analysis of 80 patients with hysterectomy specimens between NGS molecular classification and clinicopathological characters displayed that the POLE mut group was associated with EEC (87.5%) and TP53 abn subtype was correlated to a later stage (Stage II-IV, 47.6%), G3 (76.2%), serous histology (61.9%) and myometrial invasion ≥50% (47.6%). A favorable concordance (31/32, 96.9%) was shown in MSI analysis and MMR IHC results, and the agreement rate of p53 IHC and TP53 mutation was 81.5% (53/65). Compared with the p53 IHC abnormal group, the TP53 mutation group had a higher correlation with high-risk factors. A high level of concordance (31/35, 88.0%) of NGS molecular classification was achieved between curettage specimens and hysterectomy specimens while grade and histotype (including unclassified group) from curettage specimens and hysterectomy specimens showed only moderate levels of agreement, 54.3% (19/35) and 68.6% (24/35), respectively. NGS molecular classification achieved on curettage samples showed high concordance with the final hysterectomy specimens, demonstrating superior to the conventional pathological assessment of grade and histotype and potential utilization in clinical practice.


Education for practice: the role of practical experience in undergraduate and general clinical training.
A questionnaire and interview study in two London medical schools reviewed the undergraduate curricula and house year from the perspectives of graduates one year after qualification. Data from 113 house officers (44%), obtained by questionnaire and interview, showed general satisfaction with training. However, they also saw undergraduate preparation as deficient in exposure to some practical procedures and common conditions, and several difficult communication skills were not taught. Graduates reported that they would have liked more experience as undergraduates in resuscitation, lumbar puncture, inserting nasogastric tubes, endotracheal intubation, and in managing diabetic keto-acidosis, asthmatic attacks, myocardial infarcts and respiratory failure. The purposes of the house year and the articulation between it and the undergraduate years require clarification.


Evidence for the exchange of blood parasites between North America and the Neotropics in blue-winged teal (Anas discors).
Blue-winged teal (Anas discors) are abundant, small-bodied dabbling ducks that breed throughout the prairies of the northcentral USA and central Canada and that winter in the southern USA and northern Neotropics. Given the migratory tendencies of this species, it is plausible that blue-winged teal may disperse avian pathogens, such as parasites causing avian malaria, between spatially distant areas. To test the hypothesis that blue-winged teal play a role in the exchange of blood parasites between North America and areas further south, we collected information on migratory tendencies of this species and sampled birds at spatially distant areas during breeding and non-breeding periods to diagnose and genetically characterize parasitic infections. Using a combination of band recovery data, satellite telemetry, molecular diagnostics, and genetic analyses, we found evidence for (1) migratory connectivity of blue-winged teal between our sampling locations in the Canadian prairies and along the US Gulf Coast with areas throughout the northern Neotropics, (2) parasite acquisition at both breeding and non-breeding areas, (3) infection of blue-winged teal sampled in Canada and the USA with Plasmodium parasite lineages associated with the Neotropics, and (4) infection of blue-winged teal with parasites that were genetically related to those previously reported in waterfowl in both North America and South America. Collectively, our results suggest that blue-winged teal likely play a role in the dispersal of blood parasites between the Neotropics and North America, and therefore, the targeting of this species in surveillance programs for the early detection of Neotropical-origin avian pathogens in the USA may be informative.


The relationship between 'theory of mind' and attachment-related anxiety and avoidance in Italian adolescents.
This study examined the relationship between 'theory of mind' and attachment-related anxiety and avoidance in adolescence. The "Reading the Mind in the Eyes" test and the "Experiences in Close Relationships - Relationship Structures" questionnaires were administered to 402 14-19 year-old adolescents. Contrary to expectations, anxiety but not avoidance with mother was associated with less accurate mindreading, and this effect was stronger in younger than in older adolescents. Results might be explained in terms of the inconsistency of caregiver behavior that is supposed to cause anxious strategies, and thus illustrate the need to consider not only the effects, but also the causes of different types of insecure strategies.


Pediocin-like bacteriocins: new perspectives on mechanism of action and immunity.
This review attempts to analyze the mechanism of action and immunity of class IIa bacteriocins. These peptides are promising alternative food preservatives and they have a great potential application in medical sciences. Class IIa bacteriocins act on the cytoplasmic membrane of Gram-positive cells dissipating the transmembrane electrical potential by forming pores. However, their toxicity and immunity mechanism remains elusive. Here we discuss the role of the mannose phosphotransferase system (man-PTS) as the receptor for class IIa bacteriocins and the influence of the membrane composition on the activity of these antimicrobial peptides. A model that is consistent with experimental results obtained by different researchers involves the non-specific binding of the bacteriocin to the negatively charged membrane of target bacteria. This step would facilitate a specific binding to the receptor protein, altering its functionality and forming an independent pore in which the bacteriocin is inserted in the membrane. An immunity protein could specifically recognize and block the pore. Bacteriocins function in bacterial ecosystems and energetic costs associated with their production are also discussed. Theoretical models based on solid experimental evidence are vital to understand bacteriocins mechanism of action and to promote new technological developments.


Dynamic binding orientations direct activity of HIV reverse transcriptase.
The reverse transcriptase of human immunodeficiency virus (HIV) catalyses a series of reactions to convert the single-stranded RNA genome of HIV into double-stranded DNA for host-cell integration. This task requires the reverse transcriptase to discriminate a variety of nucleic-acid substrates such that active sites of the enzyme are correctly positioned to support one of three catalytic functions: RNA-directed DNA synthesis, DNA-directed DNA synthesis and DNA-directed RNA hydrolysis. However, the mechanism by which substrates regulate reverse transcriptase activities remains unclear. Here we report distinct orientational dynamics of reverse transcriptase observed on different substrates with a single-molecule assay. The enzyme adopted opposite binding orientations on duplexes containing DNA or RNA primers, directing its DNA synthesis or RNA hydrolysis activity, respectively. On duplexes containing the unique polypurine RNA primers for plus-strand DNA synthesis, the enzyme can rapidly switch between the two orientations. The switching kinetics were regulated by cognate nucleotides and non-nucleoside reverse transcriptase inhibitors, a major class of anti-HIV drugs. These results indicate that the activities of reverse transcriptase are determined by its binding orientation on substrates.


Age- and oxidative stress-induced DNA damage in Drosophila intestinal stem cells as marked by Gamma-H2AX.
A decline in stem cell function is considered as a major cause of tissue atrophy, organ-system failure, cancer development and aging process. For a better understanding of the mechanism underlying age-related decline of stem cell function, characterization of aged stem cells is required. DNA damage induces epigenetic modifications that are associated with cell dysfunction. In mammals, γH2AX has been shown as DNA damage marker and an adaptor for recruiting chromatin modifying factors. In current study, utilizing a well-accepted Drosophila midgut model for stem-cell biology, we demonstrated aging- and oxidative stress-related accumulation of γH2AvD foci, analogous to mammal γH2AX, in Drosophila intestinal stem cells (ISCs), and obtained evidence that the changes in γH2AvD is closely associated with γ-ray-induced DNA damage in ISCs and age-related accumulation of 8-oxo-2'-deoxyguanosine. The significance of our study is to document the first direct evidence for the accumulation of age-related DNA-damage in ISCs, and to show γH2AvD as a useful biomarker in exploring the molecular mechanisms underlying stem cell aging in the Drosophila midgut.


Adenosine A(2A) receptors are necessary and sufficient to trigger memory impairment in adult mice.
Caffeine (a non-selective adenosine receptor antagonist) prevents memory deficits in aging and Alzheimer's disease, an effect mimicked by adenosine A2 A receptor, but not A1 receptor, antagonists. Hence, we investigated the effects of adenosine receptor agonists and antagonists on memory performance and scopolamine-induced memory impairment in mice. We determined whether A2 A receptors are necessary for the emergence of memory impairments induced by scopolamine and whether A2 A receptor activation triggers memory deficits in naïve mice, using three tests to assess short-term memory, namely the object recognition task, inhibitory avoidance and modified Y-maze. Scopolamine (1.0 mg·kg(-1) , i.p.) impaired short-term memory performance in all three tests and this scopolamine-induced amnesia was prevented by the A2 A receptor antagonist (SCH 58261, 0.1-1.0 mg·kg(-1) , i.p.) and by the A1 receptor antagonist (DPCPX, 0.2-5.0 mg·kg(-1) , i.p.), except in the modified Y-maze where only SCH58261 was effective. Both antagonists were devoid of effects on memory or locomotion in naïve rats. Notably, the activation of A2 A receptors with CGS 21680 (0.1-0.5 mg·kg(-1) , i.p.) before the training session was sufficient to trigger memory impairment in the three tests in naïve mice, and this effect was prevented by SCH 58261 (1.0 mg·kg(-1) , i.p.). Furthermore, i.c.v. administration of CGS 21680 (50 nmol) also impaired recognition memory in the object recognition task. These results show that A2 A receptors are necessary and sufficient to trigger memory impairment and further suggest that A1 receptors might also be selectively engaged to control the cholinergic-driven memory impairment.


Paediatric Liaison Work by Child and Adolescent Mental Health Services.
Although psychiatric morbidity is common amongst paediatric patients, little is known about the availability of CAMH paediatric liaison services. We surveyed all Trusts with specialist CAMH services and paediatric units in Greater London, enquiring about the nature of liaison that CAMHS provide. We found that although liaison paediatric work was common, dedicated paediatric liaison services were provided by only a minority of specialist multidisciplinary CAMHS. Their work involved most aspects of child psychopathology, and included emergencies and children with joint physical and psychiatric problems. About 2/3 of paediatricians were satisfied with CAMHS liaison services, but virtually all desired to see them developed further. There were few indications of co-ordination between specialist multidisciplinary CAMHS and other paediatric psychosocial support services.


Immunoglobulin G antibody bound to the C1q subcomponent of human complement exhibits segmental flexibility.
The rotational dynamics of rabbit immunoglobulin G (IgG) anti-dansyl antibodies bound to the C1q subcomponent of human complement were studied by nanosecond fluorescence spectroscopy. Deconvoluted anisotropy decays of IgG-C1q mixtures were fitted to a two-exponential expression and were corrected for the effects of unbound IgG, which was determined with an analytical ultracentrifuge. Compared with the anisotropy parameters for free IgG, the pre-exponential weighting factors and the short correlation time of the C1q-bound antibody were nearly unchanged, and the long correlation time increased by only about 45 nanoseconds. These results, together with rotational diffusion calculations, indicate that the Fab arms of the C1q-bound antibody exhibited considerable flexibility. This finding may have biological relevance because it suggests that C1q can bind to the Fc segments of IgG molecules anchored in an immune complex, even though the angles between the two Fab arms of the different antibodies may vary. The results of this study also support our earlier interpretation that both the short and long correlation times of IgG principally represent flexible motions of the Fab segments.


Association of Hospital Volume and Outcomes Following Off-Pump Coronary Artery Bypass Grafting.
Off-pump coronary artery bypass grafting (OPCAB) has been used to mitigate the negative systemic effects of cardiopulmonary bypass. Recent consortium and single-institution studies suggest an association between operator experience and long-term survival. We thus aimed to ascertain the relationship between institutional OPCAB volume and outcomes using a contemporary nationwide all-payer database. Adult admissions for elective isolated OPCAB were identified from the 2016-2019 Nationwide Readmissions Database. The primary outcome was major adverse events (MAE), defined as a composite of mortality, reoperation, prolonged mechanical ventilation, acute kidney injury requiring dialysis, or perioperative stroke during the index hospitalisation. Secondary outcomes included temporal trends, postoperative length of stay (pLOS), hospitalisation costs, non-home discharge, and 30-day readmission rate. High-volume hospitals (HVH) were defined to have annual caseloads >35 based on cubic spline analysis. Of an estimated 41,154 patients, 59.9% were treated at HVH. The proportion of coronary artery bypass grafting operations that were OPCAB significantly decreased from 21.1% in 2016 to 18.3% in 2019. After adjustment, HVH status was associated with lower adjusted odds of MAE (adjusted odds ratio [AOR] 0.78, 95% confidence interval [CI] 0.70-0.88), compared to others. HVH were also associated with shorter pLOS (β -0.10, 95% -0.13, -0.07), reduced costs (β -US$4,900, - US$6,300, - US$3,600), non-home discharge (AOR 0.54, 95% CI 0.45-0.64), and 30-day readmission (AOR 0.86, 95% CI 0.77-0.96). Our results suggest that OPCAB requires a distinct set of surgical expertise and institutional aptitude. As a result, centralisation of care to centres of excellence should be considered.


c-myc oncogene protein synthesis is independent of the cell cycle in human and avian cells.
Several lines of evidence suggest a role for the myc oncogene in cell proliferation. Most recently, mitogenic stimulation of quiescent lymphoid, fibroblast and epithelial cells has been demonstrated to lead to a sharp increase in c-myc RNA levels. To determine how c-myc expression is linked to the cell proliferative cycle, we have used centrifugal elutriation to enrich for populations of avian and human cells at different stages of the cell cycle. Centrifugal elutriation is a counterflow centrifugation method that separates cells on the basis of volume, a parameter correlating well with progression through the cell cycle. Using myc-specific anti-peptide antibodies, we show here that the synthesis, half-life and modification of c-myc proteins are constant throughout the cell cycle of normal and transformed cells.


Minimal essential length of Clostridium botulinum C3 peptides to enhance neuronal regenerative growth and connectivity in a non-enzymatic mode.
C3 ADP-ribosyltransferase is a valuable tool to study Rho-dependent cellular processes. In the current study we investigated the impact of enzyme-deficient peptides derived from Clostridium botulinum C3 transferase in the context of neuronal process elongation and branching, synaptic connectivity, and putative beneficial effects on functional outcome following traumatic injury to the CNS. By screening a range of peptidic fragments, we identified three short peptides from C3bot that promoted axon and dendrite outgrowth in cultivated hippocampal neurons. Furthermore, one of these fragments, a 26-amino acid peptide covering the residues 156-181 enhanced synaptic connectivity in primary hippocampal culture. This peptide was also effective to foster axon outgrowth and re-innervation in organotypical brain slice culture. To evaluate the potential of the 26mer to foster repair mechanisms after CNS injury we applied this peptide to mice subjected to spinal cord injury by either compression impact or hemisection. A single local administration at the site of the lesion improved locomotor recovery. In addition, histological analysis revealed an increased serotonergic input to lumbar motoneurons in treated compared with control mice. Pull-down assays showed that lesion-induced up-regulation of RhoA activity within the spinal cord was largely blocked by C3bot peptides despite the lack of enzymatic activity.


Maternal homozygocity for a 14 base pair insertion in exon 8 of the HLA-G gene and carriage of HLA class II alleles restricting HY immunity predispose to unexplained secondary recurrent miscarriage and low birth weight in children born to these patients.
Homozygous carriage of a 14 base pair (bp) insertion in exon 8 of the HLA-G gene may be associated with low levels of soluble HLA-G and recurrent miscarriage (RM). We investigated the G14bp insertion(ins)/deletion(del) polymorphism in 339 women with unexplained RM and 125 control women. In all patients and patients with secondary RM after a firstborn boy, 19.2% and 23.9%, respectively, were G14bp ins/ins compared with 11.2% of controls (p<0.05 and p<0.01). Among secondary RM patients with a firstborn boy, G14bp del/del and no carriage of an HLA class II (HYrHLA) allele restricting immunity against male-specific minor HY antigens was found less often than in controls (p<0.05) whereas G14bp ins/ins and carriage of HYrHLA predisposed (p<0.08) to this clinical entity. The mean birth weight of firstborn boys born to G14bp ins positive secondary RM patients was significantly lower than expected (p<0.001) but only in carriers of HYrHLA alleles (p<0.01). In conclusion, homozygosity for G14bp ins predisposes to RM. The combination of G14 ins homozygosity and carriage of HYrHLA predisposes to secondary RM in women with a firstborn boy and negatively affects birth weight in these boys.


Longitudinal characterization of nasopharyngeal colonization with Streptococcus pneumoniae in a South African birth cohort post 13-valent pneumococcal conjugate vaccine implementation.
Monitoring changes in pneumococcal carriage is key to understanding vaccination-induced shifts in the ecology of carriage and impact on health. We longitudinally investigated pneumococcal carriage dynamics in infants. Pneumococcal isolates were obtained from nasopharyngeal (NP) swabs collected 2-weekly from 137 infants enrolled from birth through their first year of life. Pneumococci were serotyped by sequetyping, confirmed by Quellung. Pneumococci were isolated from 54% (1809/3331) of infants. Median time to first acquisition was 63 days. Serotype-specific acquisition rates ranged from 0.01 to 0.88 events/child-year and did not differ between PCV13 and non-PCV13 serotypes (0.11 events/child-year [95% CI 0.07-0.18] vs. 0.11 events/child-year [95% CI 0.06-0.18]). There was no difference in carriage duration between individual PCV13 and non-PCV13 serotypes (40.6 days [95% CI 31.9-49.4] vs. 38.6 days [95% CI 35.1-42.1]), however cumulatively the duration of carriage of non-PCV13 serotypes was greater than PCV13 serotypes (141.2 days (95% CI 126.6-155.8) vs. 30.7 days (95% CI 22.3-39.0). Frequently carried PCV13 serotypes included 19F, 9V, 19A and 6A, while non-PCV13 serotypes included 15B/15C, 21, 10A, 16F, 35B, 9N and 15A. Despite high immunization coverage in our setting, PCV13 serotypes remain in circulation in this cohort, comprising 22% of isolates. Individual PCV13 serotypes were acquired, on average, at equivalent rate to non-PCV13 serotypes, and carried for a similar duration, although the most common non-PCV13 serotypes were more frequently acquired than PCV13 serotypes.


Differences in spatial versus temporal reaction norms for spring and autumn phenological events.
For species to stay temporally tuned to their environment, they use cues such as the accumulation of degree-days. The relationships between the timing of a phenological event in a population and its environmental cue can be described by a population-level reaction norm. Variation in reaction norms along environmental gradients may either intensify the environmental effects on timing (cogradient variation) or attenuate the effects (countergradient variation). To resolve spatial and seasonal variation in species' response, we use a unique dataset of 91 taxa and 178 phenological events observed across a network of 472 monitoring sites, spread across the nations of the former Soviet Union. We show that compared to local rates of advancement of phenological events with the advancement of temperature-related cues (i.e., variation within site over years), spatial variation in reaction norms tend to accentuate responses in spring (cogradient variation) and attenuate them in autumn (countergradient variation). As a result, among-population variation in the timing of events is greater in spring and less in autumn than if all populations followed the same reaction norm regardless of location. Despite such signs of local adaptation, overall phenotypic plasticity was not sufficient for phenological events to keep exact pace with their cues-the earlier the year, the more did the timing of the phenological event lag behind the timing of the cue. Overall, these patterns suggest that differences in the spatial versus temporal reaction norms will affect species' response to climate change in opposite ways in spring and autumn.


Dimers of 5HT1 ligands preferentially bind to 5HT1B/1D receptor subtypes.
New dimers of known 5HT1 ligands (5HT, 1-NP or 8-OH-DPAT) have been prepared and evaluated at human cloned 5HT1B, 5HT1D and 5HT1A receptors. Binding experiments show that all these dimers have better affinities at 5HT1B/1D receptors than their corresponding monomeric ligands. Studies of inhibition of the forskolin-stimulated c-AMP formation mediated by the human 5HT1B receptor show that hetero-bivalent ligands [combining an agonist (5HT) with an antagonist (1-NP)] behave as partial agonists while the intrinsic activity of bivalent antagonists (combining two 1-NP residues) was found to be spacer dependent. Surprisingly enough, the dimer of 8-OH-DPAT 6 binds to 5HT1A, 5HT1B and 5HT1D receptors with similar high affinity.


Chemical modification and immunogenicity of membrane fractions from mouse tumour cells.
A crude membrane fraction isolated from mouse tumour cells was treated with various chemicals. The effects on the immunogenicity of the membrane sample were tested in syngeneic mice for tumour protection, using a challenge dose of 10(5) viable tumour cells. Best protection was obtained after immunization of mice with a membrane sample modified with dimethylsulphate. Up to 60% of the animals remained tumour free, and the tumour-bearing animals showed a greatly increased mean survival time. The post-challenge sera contained no detectable amounts of cytotoxic antibodies. The membrane sample isolated from tumour cells which had been modified with dimethylsulphate showed less immunogenicity than the modified cells or the membrane fraction from unmodified cells.


Elevated matrix metalloproteinase-2 and -3 production from human diabetic dermal fibroblasts.
Diabetic foot ulcers are characterized by elevated levels of matrix metalloproteinases (MMP), which could lead to excessive matrix breakdown and disruption to healing. It is unknown if this elevation is a function of wound healing, or if it is present within normal skin and a primary contributor to the increased risk of impaired healing. To determine whether diabetic fibroblasts from unwounded skin show elevated MMP production compared with their nondiabetic counterparts. Circular skin biopsies (4 mm diameter) were taken from the inside upper arm of four controls without diabetes and from four subjects with insulin-treated diabetes. Fibroblasts were incubated for a further 72 h and conditioned medium was collected and stored at -20 degrees C. The conditioned medium was assessed by gelatin zymography and Western blotting for MMP-2 and MMP-3. Diabetic dermal fibroblasts showed significantly elevated production of MMP-2 (P < 0.05) and pro-MMP-3 (P < 0.05) when compared with their nondiabetic counterparts. Dermal fibroblasts from normal unwounded skin are characterized by increased MMP production and this may be a primary contributing factor to the increased risk of nonhealing foot ulceration in diabetes.


Changes of vertical eye movements of goldfish for different otolith stimulation by linear acceleration.
Eye movements serves to hold the gaze steady or to shift the gaze to an object of interest. On Earth, signals from otoliths can be interpreted either as linear motion or as tilt with respect to gravity. In microgravity, static tilt will no longer give rise to changes in otolith activity. However, linear acceleration as well as angular acceleration stimulate the otolith organ. Therefore, during adaptation to microgravity, otolith-mediated response such as eye movements alter. In this study, we analyzed the eye movements of goldfish during linear acceleration. The eye movements during rectangular linear acceleration along the different body axis were video-recorded. The vertical eye rotations were analyzed frame by frame. In normal fish, leftward lateral acceleration induced downward eye rotation in the left eye and upward eye rotation in the right eye. Acceleration from caudal to rostral evoked downward eye rotation in both eyes. When the direction of acceleration was shifted 15 degrees left, the responses in the left eye disappeared. These results suggested that otolith organs in each side were stimulated differently.


LAT1-specific inhibitor ameliorates severe autoimmune arthritis in SKG mouse.
L-type amino acid transporter 1 (LAT1, slc7a5) supplies large neutral amino acids to highly proliferative cells. LAT1 is an attractive therapeutic target for treating overactive T cell-mediated immune disorders due to its high expression in activated T cells, but not in resting T cells. Here, we demonstrate that LAT1 plays a crucial role in T helper (Th) 17-mediated autoimmune arthritis in SKG mice, an animal model of human rheumatoid arthritis (RA). Administration of JPH203, a LAT1-specific inhibitor, suppressed mannan-induced joint swelling, synoviocyte proliferation and inflammatory cell infiltration in SKG mice. A diminished metabolic reprogramming, including a decrease in oxidative phosphorylation that regulates Hif-1α expression and subsequent control of glycolysis enzymes, was involved in the downregulation of Th17 differentiation by LAT1 inhibition. Moreover, publicly released database analysis revealed facilitated expression of LAT1 in T cells with cytotoxic features in patients with RA. Our results demonstrate the essential contribution of LAT1 to the development of RA, proposing a potential therapeutic approach targeting amino acid transporters for treating hypersensitive immune diseases.


Tracking spoilage bacteria in commercial poultry processing and refrigerated storage of poultry carcasses.
Four trials were conducted to examine the effect of commercial processing and refrigerated storage on spoilage bacteria in the native microflora of broiler carcasses. Prescalded, picked, eviscerated, and chilled carcasses were obtained from a commercial processing facility, and psychrotrophs in the bacterial flora were enumerated on Iron Agar, Pseudomonas Agar, and STAA Agar. The size of the population of spoilage bacteria on processed carcasses stored at 4 degrees C for 7, 10, or 14 days was also determined. Bacterial isolates were identified and dendrograms of the fatty acid profiles of the isolates were prepared to determine the degree of relatedness of the isolates. Findings indicated that although some processing steps increased the level of carcass contamination by selected bacteria, the number of spoilage bacteria recovered from processed carcasses was significantly (P< or = 0.05) less than the number of bacteria recovered from carcasses entering the processing line. Acinetobacter and Aeromonas spp. were the primary isolates recovered from carcasses taken from the processing line. During refrigerated storage, there was a significant (P < or =0.05) increase in the population of bacteria on the carcasses, and Pseudomonas spp. were the predominant bacteria recovered from these carcasses. Dendrograms of the fatty acid profiles of the isolates indicated that bacterial cross-contamination of carcasses occurs during all stages of processing and that some bacteria can survive processing and proliferate on carcasses during refrigerated storage. Furthermore, cross-contamination was detected between carcasses processed on different days at the same facility. Findings indicate that although poultry processing decreases carcass contamination by psychrotrophic spoilage bacteria, significant levels of bacterial cross-contamination occur during processing, and bacteria that survive processing may multiply on the carcasses during refrigerated storage.


Animal models for human T-lymphotropic virus type 1 (HTLV-1) infection and transformation.
Over the past 25 years, animal models of human T-lymphotropic virus type 1 (HTLV-1) infection and transformation have provided critical knowledge about viral and host factors in adult T-cell leukemia/lymphoma (ATL). The virus consistently infects rabbits, some non-human primates, and to a lesser extent rats. In addition to providing fundamental concepts in viral transmission and immune responses against HTLV-1 infection, these models have provided new information about the role of viral proteins in carcinogenesis. Mice and rats, in particular immunodeficient strains, are useful models to assess immunologic parameters mediating tumor outgrowth and therapeutic invention strategies against lymphoma. Genetically altered mice including both transgenic and knockout mice offer important models to test the role of specific viral and host genes in the development of HTLV-1-associated lymphoma. Novel approaches in genetic manipulation of both HTLV-1 and animal models are available to address the complex questions that remain about viral-mediated mechanisms of cell transformation and disease. Current progress in the understanding of the molecular events of HTLV-1 infection and transformation suggests that answers to these questions are approachable using animal models of HTLV-1-associated lymphoma.


In-vivo biosynthesis of acid glycosaminoglycans with radiosulfate and [3H]glucosamine in single joints from rats of different age.
Administration intraperitoneally of 35SO4 plus D-[6-3H]glucosamine of similarly specific radioactivity to rats of different ages showed uptake rates and incorporation to be higher with 35SO4 in articular cartilage. Comparing the joints studied: knee joints (femur and tibia parts), humerus, and femur heads, cartilage cells appeared to be most metabolically active (age-dependently) in femur heads. All cells, in addition to glycoproteins, synthesized in vivo isomeric chondroitin sulfates and hyaluronate at different rates.


Familial glioma: a report of glioblastoma in identical twins and oligo-astrocytoma in siblings.
The authors report two pairs of siblings who had intracranial gliomas. One pair of identical twins had glioblastomas and two siblings had mixed oligo-astrocytomas. Genetic influences in the aetiology of glioma are discussed.


PAINSTORIES - Exploring the Temporal Developments in the Challenges, Barriers, and Self-Management Needs of Adolescents with Longstanding Knee Pain: A Qualitative, Retrospective Interview Study with Young Adults Experiencing Knee Pain Since Adolescence.
Patient education is essential to enable rehabilitation and self-management of longstanding knee pain in adolescents. Currently, a lack of insights into the socio-cognitive processes governing adolescents' self-management remains an obstacle for enhancing treatment efficacy. This study developed a conceptual model for integrating adolescents' challenges and barriers into future treatments. We conducted semi-structured retrospective interviews with 14 young adults (age 21-25 years) with knee pain since adolescence (9 years mean duration). Temporal developments in participants' knee pain were captured through a memorization exercise. Data was analyzed via the General Inductive Approach. Themes were organized into a matrix, extracting a conceptual model, which was tested with eight new participants. The analysis identified seven themes. Further interpretation, via the matrix, organized these within a four-stage trajectory of; gaining awareness, knowledgeability, contextual application and reconceptualization, each with different challenges and dilemmas, participants had to overcome to progress their self-management. Testing the conceptual model, confirmed stages and highlighted acceptance as key to overcoming barriers. The study described adolescents' integration of self-management as proximal and inquiry-based, with acceptance, driving increasingly complex management behaviors. We hypothesize future interventions may benefit from exploring supporting adolescents' inquiries into their knee pain at different stages of the trajectory. PERSPECTIVE: This study presents a conceptual model and vocabulary for optimizing patient education concepts, to target the challenges, barriers and needs of adolescents with knee pain at different stages of their mastery journey. We believe our findings may inform reflections among clinicians and researchers, and development of more effective education interventions.


Hemiplegic writing in severe aphasia.
Three patients with severe aphasia and right hemiplegia are described who could write to dictation with the right arm using a limb prosthesis though agraphic with the "intact" left hand. The phenomenon of "hemiplegic writing" is explained as an access to submerged or preprocessing levels in language and action structure, through the use of older proximal motor systems. This interpretation has implications for our understanding of language and brain function, as well as for approaches to the treatment of patients with severe language disorders.


Age-related changes in contextual associative learning.
The hippocampus plays a critical role in processing contextual information. Although age-related changes in the hippocampus are well documented in humans, nonhuman primates, and rodents, few studies have examined contextual learning deficits in old rats. The present study investigated age-related differences in contextual associative learning in young (6 mo) and old (24 mo) rats using olfactory stimuli. Stimuli consisted of common odors mixed in sand and placed in clear plastic cups. Testing was conducted in two boxes that represented two different contexts (Context 1 and Context 2). The contexts varied based on environmental features of the box such as color (black vs. white), visual cues on the walls of the box, and flooring texture. Each rat was simultaneously presented with two cups, one filled with Odor A and one filled with Odor B in each context. In Context 1, the rat received a food reward for digging in the cup containing Odor A, but did not receive a food reward for digging in the cup containing Odor B. In Context 2, the rat was rewarded for digging in the cup containing Odor B, but did receive a reward for digging in the cup containing Odor A. Therefore, the rat learned to associate Context 1 with Odor A and Context 2 with Odor B. The rat was tested for eight days using the same odor problem throughout all days of testing. The results showed no significant difference between young and old rats on the first two days of testing; however, young rats significantly outperformed old rats on Day 3. Young rats continued to maintain superior performance compared to old rats on Days 4-8. The results suggest that aging results in functional impairments in brain regions that support memory for associations between specific cues and their respective context.


[Bilateral unipedicle flaps combined with levator sling plasty for the repair of complete cleft palate at the age of 6 - 12 months].
To repair the complete cleft palate with the most popular technique at optimal time. Bilateral unipedicle flaps (Bardach method) combined with levator sling plasty were employed to repair complete cleft palate at the age of 6 - 12 months. Computer-aided FFT vocal analysis was performed before and after surgery. All patients had primary wound healing without any complication. The FFT vocal analysis showed great improvement in velopharyngeal incompetence after surgery. It is technically safe and feasible to repair the complete cleft palate at the age of 6 - 12 months with bilateral unipedicle flaps. Encouraging speech improvement can be expected with this method.


Inhibition of the motility of Artemia salina L. (brine shrimps) by keto-trichothecenes.
Four keto-trichothecenes (nivalenol, deoxynivalenol, 3-acetyl-deoxynivalenol, and fusarenone-X) were tested in a new "brine shrimp" bioassay, showing that NIV, DON, and F-X are highly active in this system. An improved equipment for the breeding and separating of larvae is indicated.


A hydrophilic interaction chromatography-tandem mass spectrometry method for amino acid profiling in mussels.
A UHPLC-HILIC-tandem MS method has been developed and validated for the quantification of 21 amino acids (20 protein amino acids and cystine) in their free form (FAA) and as protein constituents (total amino acids, TAA) in a rich protein food matrix such as lyophilized mussels (Mytilus galloprovincialis) samples. FAA were analyzed after suspending the samples in the presence of trichloroacetic acid in order to prevent dissolving the proteins, while TAA were determined after acid hydrolysis with 6M HCl in the presence of 4% v/v thioglycolic acid as a reducing agent. In hydrolysed samples 17 amino acids could be determined since tryptophan, cysteine, cystine and asparagine were degraded during acid hydrolysis. Linear regression coefficients (R2) were above 0.99 for all amino acids. Accuracy and precision, expressed as recovery (%) and relative standard deviation (RSD, %) were in acceptable levels, ranging from 78.2 to 123.3% and below 15%, respectively for both FAA and TAA. Uncertainty was also below 12% for FAA and below 22% for TAA. Sensitivity of the method was high with LOD values ranging from 0.003 to 0.034g/100g for FAA and 0.001 to 0.004g/100g for TAA, while LOQ ranged from 0.009 to 0.104g/100g for FAA and 0.002 to 0.011g/100g for TAA. The method proved to be a fast and reliable tool for acquiring information on free and total amino acids profile in high protein content foodstuffs such as mussels.


Nitric oxide production and monoamine oxidase activity in cancer patients during interferon-alpha therapy.
Both increased and decreased nitric oxide (NO) synthesis have been reported in patients treated with interferon-alpha (IFN-alpha). Animal studies showed that IFN-alpha administration results in increased levels of biogenic amines, subsequent activation of monoamine oxidases (MAOs), and finally in a change in NO production due to the H(2)O(2) generated by MAOs. We examined the potential relationship between NO production in plasma and MAO-B activity in platelets of 43 cancer patients during 8 weeks of treatment with IFN-alpha. NO synthesis was quantitated by measuring both the ratio of citrulline and arginine (CIT/ARG-ratio) and total nitrite/nitrate (NOx) levels. Compared to baseline, MAO activity and NOx increased, while the CIT/ARG-ratio decreased. No associations were found between NOx, MAO and CIT/ARG-ratio. Only few associations were observed between changes in the biochemical parameters and changes in psychopathology induced by IFN-alpha, of which the association between changes in CIT and lassitude was the most consistent. The results suggest that peripheral NO production and MAO activity are unrelated to each other, and that peripheral changes in these biochemical parameters induced by IFN-alpha are unlikely to contribute to definite psychiatric disturbance.


Nicotinamide-adenine dinucleotide inhibition of pig kidney alkaline phosphatase.
1. The interaction of NAD+, NADH and various nucleotide analogues with pig kidney alkaline phosphatase (orthophosphoric-monoester phosphohydrolase (alkaline optimum) EC 18.104.22.168) has been investigated by kinetic means. Some inhibitors act uncompetitively whereas others markedly increase the slopes of double reciprocal plots suggesting they have some affinity for the free enzyme. 2. The compounds seem to bind to alkaline phosphatase through interactions of their bases with a relatively non-specific region of the enzyme, although it is likely that for those nucleotides having some affinity for the free enzyme there is some attraction between the pyrophosphate backbone and the active site. 3. From studies of the effect of NAD+ and NADH on ATPase activity it was concluded that the substrate inhibition that is characteristic of the ATPase activity of alkaline phosphatase originates from binding of ATP to the site assumed to exist for NAD+ and NADH. The potentiation of NAD+-inhibition of ATPase activity by Mg-2+ is probably a result of the depletion of [ATP-4-] the true substrate. The depletion allows NAD+ to complete more effectively for the active site. 4. Binding of NADH is favoured by protonation of an enzymic group with a pK of approx. 9.0 belonging possibly to a tyrosine residue or a zinc hydrate. 5. A large entropy decrease was found to accompany the binding of NAD+ and NADH to alkaline phosphatase. This may be further evidence of an "induced-fit" mechanism previously suspected because of the synergistic inhibitory effects of adenosine and nicotinamide.


Cuprolinic Blue: a specific dye for single-stranded RNA in the presence of magnesium chloride. II. Practical applications for light microscopy.
The application of the new nucleic acid dye Cuprolinic Blue to cell smears and tissue sections has been described. Without added cations, Cuprolinic Blue stains both DNA and RNA, whereas in the presence of 1 M MgCl2, Cuprolinic Blue specifically stains single-stranded RNA only. The total RNA can be stained after removal of DNA by DNAase digestion. Fixation in a modified Carnoy solution gave optimal staining results in all cases tested. By cytophotometry, a reliable and reproducible relative estimate can be obtained of the total nucleic acid content, the total RNA content and the amount of single-stranded RNA alone per cell.


Partitioning of polychlorinated biphenyls and hexachlorobenzene into human faeces.
The dietary absorption of persistent lipophilic organic pollutants (PLOPs) in humans is believed to occur via partitioning of the chemical between the lumen and the wall of the digestive tract. As such, the partitioning properties of the lumen contents are a key factor governing absorption. In this study, the partitioning properties of faeces were measured for 11 polychlorinated biphenyls and hexachlorobenzene (HCB). Four volunteers participated in the study, each of them providing faeces from a normal diet and a vegetarian diet. The faeces/gas equilibrium partition coefficient K(FG) varied by over three orders of magnitude between the different compounds. A linear relationship between log K(FG) and log KOA, the octanol/air partition coefficient, was observed. The slope of the relationship was > 1, indicating that the solvent properties of faeces were less polar than those of octanol. For a given compound, KFG varied up to a factor of 2.8 between the individuals on a normal diet. The influence of the vegetarian diet on K(FG) was negligible for the two volunteers who simply deleted fish and animal products from their normal diet, but K(FG) increased on average by a factor of 2 in the two individuals who increased their consumption of less readily digestible whole grains and vegetables in their vegetarian diet. On the basis of K(FG), the fugacities in the faeces were calculated. They were found to be much lower than the fugacities in blood. It is hypothesised that this is due to a temporary decrease in the fugacity in the wall of the jejunum caused by absorption of dietary lipids that results in equilibration between the lumen contents and the wall of the digestive tract at a fugacity below that present in the blood and the rest of the body.


Effects of an eight week military training program on aerobic indices and psychomotor function.
This study assessed the effects of eight weeks of military training on aerobic fitness indices, military skills and neuropsychological function. Thirty five (n = 35) male Irish Defence Forces personnel, divided into training (n = 20) and control (n = 15) subgroups, completed tests of military aptitude (Kim's games, judging distance, fire order, map reading, weapon assembly) and neuropsychological function (Symbol digit modalities test (SDMT), Trail making test, Stroop test and grooved pegboard test) pre- and post-intervention. The repeated measures study design sought to account for any learning effect. Participants also completed a 10km route march, a two mile run and three by 20m shuttle run tests at both time points to quantify changes in fitness variables. The training sub-group significantly (P < 0.001) improved mean 20m shuttle-run distance and consequently estimated VO2 max pre- to post-intervention (49.8 +/- 1.0 vs. 52.4 +/- 0.9 mL x kg x min(-1)). Two mile run time was not significantly improved. Mean %HRmax during the 10km route march was significantly higher in both training (P < 0.001) and control (P < 0.01) sub-groups post-intervention (71 +/- 1 and 83 +/- 1%) compared to pre-intervention (65 +/- 1 and 77 +/- 1%). However, the training sub-group conducted the route march at a significantly faster speed on the second occasion. Military training significantly improved performance in 3/18 neuropsychological test components and 2/12 military skills test components. Training significantly improved ability to estimate both short (error; 36 +/- 6 vs. 12 +/- 1%) and intermediate (error; 72 +/- 12 vs. 11 +/- 3%) distances post-intervention. The training sub-group significantly (P < 0.01) improved SDMT score and mean Trail 1 time pre- to post-intervention (58.0 +/- 2.8 vs. 69.5 +/- 3.4; 18.1 +/- 0.8 vs. 14.4 +/- 0.8s, respectively). In Part 3 of the Stroop test, time mediated a significant (P < 0.05) and selective improvement in the training sub-group (51.3 +/- 3.2 vs. 63.8 +/- 5.4). In conclusion, aerobic fitness and a minority of neuropsychological and military skills tests improved following 8 weeks of military training.


Differentiation between two "fang ji" herbal medicines, Stephania tetrandra and the nephrotoxic Aristolochia fangchi, using hyperspectral imaging.
Stephania tetrandra ("hang fang ji") and Aristolochia fangchi ("guang fang ji") are two different plant species used in Traditional Chinese Medicine (TCM). Both are commonly referred to as "fang ji" and S. tetrandra is mistakenly substituted and adulterated with the nephrotoxic A. fangchi as they have several morphological similarities. A. fangchi contains aristolochic acid, a carcinogen that causes urothelial carcinoma as well as aristolochic acid nephropathy (AAN). In Belgium, 128 cases of AAN was reported while in China, a further 116 cases with end-stage renal disease were noted. Toxicity issues associated with species substitution and adulteration necessitate the development of reliable methods for the quality assessment of herbal medicines. Hyperspectral imaging in combination with partial least squares discriminant analysis (PLS-DA) is suggested as an effective method to distinguish between S. tetrandra and A. fangchi root powder. Hyperspectral images were obtained in the wavelength region of 920-2514nm. Reduction of the dimensionality of the data was done by selecting the discrimination information range (964-1774nm). A discrimination model with a coefficient of determination (R(2)) of 0.9 and a root mean square error of prediction (RMSEP) of 0.23 was created. The constructed model successfully identified A. fangchi and S. tetrandra samples inserted into the model as an external validation set. In addition, adulteration detection was investigated by preparing incremental adulteration mixtures of S. tetrandra with A. fangchi (10-90%). Hyperspectral imaging showed the ability to accurately predict adulteration as low as 10%. It is evident that hyperspectral imaging has tremendous potential in the development of visual quality control methods which may prevent cases of aristolochic acid nephropathy in the future.


Geographic dark posterior fundus patches.
Nine cases of a newly recognized, probably not uncommon, anomaly of the retinal pigment epithelium have been observed over a four-year period. The lesions occur predominantly in black subjects. Some degree of hemoglobinopathy was present in all the black patients with geographic dark posterior fundus patches. Almost all of the patients were young (6 to 26 years old) but two were 50 years of age. In three patients the patches grew larger. They did not interfere with vision or visual fields and did not appear to be involved with vitreous abnormalities. Fluorescein angiography was normal. In three patients, an electroretinogram gave subnormal results to all stimulus intensities; it was normal in one patient.


Calmodulin regulation of adenylate cyclase activity.
Calmodulin-dependent stimulation of adenylate cyclase was initially thought to be a unique feature of neural tissues. In recent years evidence to the contrary has accumulated, calmodulin-dependent stimulation of adenylate cyclase now being demonstrated in a wide range of structurally unrelated tissues and species. Demonstration of the existence of calmodulin-dependent adenylate cyclase has in nearly all instances required the removal of endogenous calmodulin. It is not yet clear whether calmodulin-dependent and calmodulin-independent forms of the enzyme exist and whether some tissues (such as heart) lack a calmodulin-dependent adenylate cyclase. The presence of calmodulin appears largely responsible for the ability of the adenylate cyclase enzyme to be stimulated by submicromolar concentrations of calcium; it may not be relevant to the inhibition of the enzyme which occurs at higher concentrations of calcium. The physical relationship of calmodulin to the plasma membrane bound enzyme (or to the soluble forms of the enzyme) is not known nor is the mechanism of adenylate cyclase activation by calmodulin clear; current data suggest some involvement with both the N and C units of the enzyme. Finally, it is possible that in vivo calcium contributes to the duration of the hormone stimulated cyclic AMP signal. Thus current in vitro data suggest that optimal hormonal activation of calmodulin-dependent adenylate cyclase occurs at very low intracellular calcium concentrations, comparable to those found in the resting cell; conversely the enzyme is inhibited as intracellular calcium increases, following for example agonist stimulation of the cell. These higher calcium concentrations would then activate calmodulin-dependent phosphodiesterase. Such differential effects of calcium on adenylate cyclase and phosphodiesterase would ultimately restrict the duration of the hormone-induced cyclic AMP signal.


Age- and gender-specific normative data on computerized dynamic posturography in a cohort of Danish adults.
Balance performance can be measured with a computerized dynamic posturography (CDP). Frequently used CDP protocols include the sensory organization test (SOT), motor control test (MCT) and adaptation test (ADT). The primary aim of this study was to produce normative data using the Bertec® Balance Advantage CDP on a cohort of healthy adults aged 20-79 years for the SOT, MCT and ADT. The secondary aim was to determine age- and gender-related differences. The study population of 120 participants were divided into age groups of 20-29, 30-39 years, etc., with an equal number of men and women in each group. Using the Bertec® CDP, the SOT, MCT and ADT protocols were performed. SOT found that the 70-79 age group and men scored lower equilibrium scores (ES) on SOT4, and SOT6. Women had higher ES on SOT1. For the MCT, backward translation. latencies were higher for the 60 s and 70 s age groups. The 60 s age group had higher forward translation latencies. Women had lower latencies on all MCTs. For the ADT toes up test, the 70-79 age group scored higher sway energy. On the toes down test, higher sway energy was registered for the 50-79 age groups. Women had lower sway energy for both ADT's. We have produced normative data for healthy adults aged 20-79 years for the SOT, MCT and ADT protocols using the Bertec® Balance Advantage CDP. Balance performance differences might be due to decline in visual acuity, presbyvestibulopathy and differences in body composition.


Porcine deltacoronavirus: Overview of infection dynamics, diagnostic methods, prevalence and genetic evolution.
Porcine deltacoronavirus (PDCoV) was first reported in Hong Kong, China in 2012 and reported in United States swine in February 2014. PDCoV has subsequently been detected in South Korea, mainland China, and Thailand. PDCoV has been experimentally confirmed to cause diarrhea in inoculated pigs and need to be differentially diagnosed from porcine epidemic diarrhea virus and transmissible gastroenteritis virus in the field. Rapid diagnosis is critical for the implementation of efficient control strategies against PDCoV. Developing high-quality diagnostic methods and understanding PDCoV infection dynamics to collect appropriate specimens at the appropriate time window are important to obtain reliable diagnostic results. Among the virological methods, PDCoV-specific RT-PCR remains the method of choice for the detection of PDCoV; immunohistochemistry combined with hematoxylin and eosin staining has also been commonly used to examine histopathological lesions caused by PDCoV. Serological assays can provide information about previous exposure to PDCoV and also determine antibody responses to infection or vaccination. Prevalence of PDCoV is lower compared to that of PEDV. However, among PDCoV-positive samples, co-infection with other enteric pathogen e.g. PEDV is common. It is also important to understand molecular epidemiology of PDCoV and genetic relationships of global PDCoVs. This review discusses PDCoV infection dynamics and appropriate sample collection for diagnostic testing, the commonly used virological and serological methods for PDCoV diagnosis, prevalence and genetic evolution of PDCoVs.


Meta-Analysis on the Association Between Nutritional Status and Outcomes After Transcatheter Aortic Valve Implantation.
Malnutrition is a common co-morbidity among candidates for transcatheter aortic valve implantation (TAVI). This study aimed to investigate the association between nutritional status determined by objective nutritional indices and outcomes of patients who underwent TAVI. We systematically searched PubMed, Embase, Web of Science, Scopus, and Cochrane Library from inception until April 18, 2022 to identify studies examining the association of preprocedural nutritional status with post-TAVI outcomes. Malnutrition was defined by objective nutritional indices-controlling nutritional index, nutritional risk index, geriatric nutritional risk index (GNRI), and prognostic nutritional index (PNI). The primary end point was 1-year all-cause mortality. The review included 13 observational studies and 6,785 patients who underwent TAVI. Malnutrition was associated with a higher risk of 1-year all-cause mortality, as defined by either the controlling nutritional index (hazard ratio [HR] 2.70, 95% confidence interval [CI] 1.21 to 6.03, p = 0.015), GNRI (HR 1.79, 95% CI 1.09 to 2.93, p = 0.021), or PNI (HR 1.17, 95% CI 1.11 to 1.23, p <0.001). In the meta-analysis of adjusted results, lower GNRI was independently associated with higher 1-year mortality (HR 1.70, 95% CI 1.16 to 2.50, p = 0.006). Lower GNRI was associated with increased risk of acute kidney injury (relative risk [RR] 2.21, 95% CI 1.63 to 2.99, p <0.001) and 1-year cardiovascular mortality (RR 2.50, 95% CI 1.66 to 3.78, p <0.001). Lower PNI was associated with a higher risk of major vascular complications (RR 2.99, 95% CI 1.38 to 6.51, p = 0.006). In conclusion, baseline malnutrition, as assessed by objective indices, is associated with worse outcomes after TAVI. Future studies should focus on the value of nutritional assessment and interventions to improve nutritional status in patients who underwent TAVI.