Effects of rogue ryanodine receptors on Ca2+ sparks in cardiac myocytes.
Ca2+ sparks and Ca2+ quarks, arising from clustered and rogue ryanodine receptors (RyRs), are significant Ca2+ release events from the junctional sarcoplasmic reticulum (JSR). Based on the anomalous subdiffusion of Ca2+ in the cytoplasm, a mathematical model was developed to investigate the effects of rogue RyRs on Ca2+ sparks in cardiac myocytes. Ca2+ quarks and sparks from the stochastic opening of rogue and clustered RyRs are numerically reproduced and agree with experimental measurements. It is found that the stochastic opening Ca2+ release units (CRUs) of clustered RyRs are regulated by free Ca2+ concentration in the JSR lumen (i.e. [Ca2+]lumen). The frequency of spontaneous Ca2+ sparks is remarkably increased by the rogue RyRs opening at high [Ca2+]lumen, but not at low [Ca2+]lumen. Hence, the opening of rogue RyRs contributes to the formation of Ca2+ sparks at high [Ca2+]lumen. The interplay of Ca2+ sparks and Ca2+ quarks has been discussed in detail. This work is of significance to provide insight into understanding Ca2+ release mechanisms in cardiac myocytes.


Developmental learning in a pain-related system: evidence for a cross-modality mechanism.
The nociceptive spinal reflex system performs highly precise sensorimotor transformations that require functionally specified synaptic strengths. The specification is gradually attained during early development and appears to be learning dependent. Here we determine the time course of this specification for heat-nociceptive tail withdrawal reflexes and analyze which types of primary afferents are important for the learning by applying various forms of noninvasive sensory deprivations. The percentage of erroneous heat-nociceptive tail withdrawal reflexes (i.e., movements directed toward the stimulation) decreased gradually from 64.1 +/- 2.5% (mean +/- SEM) to <10% during postnatal days 10-21. This improvement was completely blocked by anesthetizing the tail during the adaptation period, confirming that an experience-dependent mechanism is involved in the specification of synaptic strengths. However, the results show that the adaptation occurs to a significant extent despite local analgesia and protection of the tail from noxious input, provided that tactile sensitivity is preserved. Therefore, it appears that a nociceptive input is not necessary for the adaptation, and that input from tactile receptors can be used to guide the nociceptive synaptic organization during development. Sensory deprivation in the adult rat failed to affect the heat-nociceptive withdrawal reflex system, indicating that the adaptation has a "critical period" during early development. These findings provide a key to the puzzle of how pain-related systems can be functionally adapted through experience despite the rare occurrence of noxious input during early life.


Neuropeptides as possible targets in sleep disorders: special emphasis on hypocretin-deficient narcolepsy.
Sleep disorders are disturbances of usual sleep patterns or behaviors caused by deregulation of neuronal synchronicity and of the balance of the neurotransmitter system involved in sleep regulation. Insomnia and hypersomnia are frequent sleep disorders, and these are most often treated pharmacologically with hypnotics and wake-promoting compounds. These compounds act on classical neurotransmitter systems, such as benzodiazepines on gamma amino butyric acid (GABA)(A) receptors, and amphetamine-like stimulants on monoaminergic terminals to modulate neurotransmission. In addition, acetylcholine, amino acids, lipids and proteins (cytokines) and peptides, are known to significantly modulate sleep, and thus, are possibly involved in the pathophysiology of some sleep disorders. Due to recent developments in molecular biological techniques, many neuropeptides have been newly identified, and some are found to significantly modulate sleep. Recent discoveries also include the finding that the impairment of hypocretin/orexin neurotransmission (a recently isolated hypothalamic neuropeptide and receptor system), is the major pathophysiology of narcolepsy with cataplexy. A hypocretin replacement therapy is anticipated to reverse the disease symptoms in humans. In this article, we will review the history of neuropeptide research, sleep modulatory effects of various neuropeptides, and the general strategies for the pharmacological therapeutics targeting the peptidergic systems by referring to hypocretin-deficient narcolepsy as an immediate example.


KI-67 heterogeneity in well differentiated gastro-entero-pancreatic neuroendocrine tumors: when is biopsy reliable for grade assessment?
Ki-67 heterogeneity can impact on gastroenteropancreatic neuroendocrine tumor grade assignment, especially when tissue is scarce. This work is aimed at devising adequacy criteria for grade assessment in biopsy specimens. To analyze the impact of biopsy size on reliability, 360 virtual biopsies of different thickness and lengths were constructed. Furthermore, to estimate the mean amount of non-neoplastic tissue component present in biopsies, 28 real biopsies were collected, the non-neoplastic components (fibrosis and inflammation) quantified and the effective area of neoplastic tissue calculated for each biopsy. Heterogeneity of Ki-67 distribution, G2 tumors and biopsy size all play an important role in reducing the reliability of biopsy samples in Ki-67-based grade assignment. In particular in G2 cases, 59.9% of virtual biopsies downgraded the tumor and the smaller the biopsy, the more frequent downgrading occurs. In real biopsies the presence of non-neoplastic tissue reduced the available total area by a mean of 20%. By coupling the results from these two different approaches we show that both biopsy size and non-neoplastic component must be taken into account for biopsy adequacy. In particular, we can speculate that if the minimum biopsy area, necessary to confidently (80% concordance) grade gastro-entero-pancreatic neuroendocrine tumors on virtual biopsies ranges between 15 and 30 mm2, and if real biopsies are on average composed of only 80% of neoplastic tissue, then biopsies with a surface area not <12 mm2 should be performed; using 18G needles, this corresponds to a minimum total length of 15 mm.


Overcoming differential tumor penetration of BRAF inhibitors using computationally guided combination therapy.
BRAF-targeted kinase inhibitors (KIs) are used to treat malignancies including BRAF-mutant non-small cell lung cancer, colorectal cancer, anaplastic thyroid cancer, and, most prominently, melanoma. However, KI selection criteria in patients remain unclear, as are pharmacokinetic/pharmacodynamic (PK/PD) mechanisms that may limit context-dependent efficacy and differentiate related drugs. To address this issue, we imaged mouse models of BRAF-mutant cancers, fluorescent KI tracers, and unlabeled drug to calibrate in silico spatial PK/PD models. Results indicated that drug lipophilicity, plasma clearance, faster target dissociation, and, in particular, high albumin binding could limit dabrafenib action in visceral metastases compared to other KIs. This correlated with retrospective clinical observations. Computational modeling identified a timed strategy for combining dabrafenib and encorafenib to better sustain BRAF inhibition, which showed enhanced efficacy in mice. This study thus offers principles of spatial drug action that may help guide drug development, KI selection, and combination.


Hepatitis B virus DNA detected in formalin-fixed liver specimens and its relation to serologic markers and histopathologic features in chronic liver disease.
To examine the relationship of hepatitis B virus (HBV) DNA sequences in the liver with histopathologic features and antigenic markers, the authors determined the hepatocytic status of viral DNA by in situ hybridization in formalin-fixed liver sections using a biotinylated probe in 45 patients with various chronic liver diseases. The results were compared retrospectively with the HBV serologic markers and histopathologic features including the presence of ground-glass cells or Shikata staining positivity. The specificity of this in situ detection of HBV DNA has been proven excellent in a double-blind control study in 18 patients in whom liver HBV DNA was also determined by DNA extraction, gel electrophoresis, and the Southern blotting technique. In 41 patients, the findings of HBV DNA and serologic markers were concordant (17 positive and 24 negative). Twelve of the 20 HBV-DNA-positive patients were HBsAg-positive (6 with chronic hepatitis, 3 with cirrhosis, and 3 with hepatocellular carcinoma). Ground-glass cells or Shikata positivity were found in 10 of these 12 patients. HBV DNA sequences were found in the liver of all patients with chronic liver disease and serologic positivity for HBV infection. In liver with normal histologic features, HBV DNA was not demonstrable, despite the positive anti-HBc and anti-HBs. However, a positive HBV DNA was found in 3 serologically negative patients. In another patient the interpretation of findings was impossible because of severe hemosiderosis. From this study, it is concluded that in situ detection of HBV DNA in formalin-fixed liver sections has a clinical value and is suitable for routine use.


Early tracheal extubation after coronary artery bypass graft surgery reduces costs and improves resource use. A prospective, randomized, controlled trial.
Economics has caused the trend of early tracheal extubation after cardiac surgery, yet no prospective randomized study has directly validated that early tracheal extubation anesthetic management decreases costs when compared with late extubation after cardiac surgery. This prospective, randomized, controlled clinical trial was designed to evaluate the cost savings of early (1-6 h) versus late tracheal extubation (12-22 h) in patients after coronary artery bypass graft (CABG) surgery. The total cost for the services provided for each patient was determined for both the early and late groups from hospital admission to discharge home. All costs applicable to each of the services were classified into direct variables, direct fixed costs, and overhead (an indirect cost). Physician fees and heart catheterization costs were included. The total service cost was the sum of unit workload and overhead costs. One hundred patients having elective CABG who were younger than 75 yr were studied. Including all complications, early extubation (n = 50) significantly reduced cardiovascular intensive care unit (CVICU) costs by 53% (P < 0.026) and the total CABG surgery cost by 25% (P < 0.019) when compared with late extubation (n = 50). Forty-one patients (82%) in each group were tracheally extubated within the defined period. In the early extubation group, the actual departmental cost savings in CVICU nursing and supplies was 23% (P < 0.005), in ward nursing and supplies was 11% (P < 0.05), and in respiratory therapy was 12% (P < 0.05). The total cost savings per patient having CABG was 9% (P < 0.001). Further cost savings using discharge criteria were 51% for CVICU nursing and supplies (P < 0.001), 9% for ward nursing and supplies (P < 0.05), and 29% for respiratory therapy (P < 0.001), for a total cost savings per patient of 13% (P < 0.001). Early extubation also reduced elective case cancellations (P < 0.002) without any increase in the number of postoperative complications and readmissions. Early tracheal extubation anesthetic management reduces total costs per CABG surgery by 25%, predominantly in nursing and in CVICU costs. Early extubation reduces CVICU and hospital length of stay but does not increase the rate or costs of complications when compared with patients in the late extubation group. It shifts the high CVICU costs to the lower ward costs. Early extubation also improves resource use after cardiac surgery when compared with late extubation.


Coiled-coil coactivators play a structural role mediating interactions in hypoxia-inducible factor heterodimerization.
The hypoxia-inducible factor complex (HIF-α·aryl hydrocarbon receptor nuclear translocator (ARNT)) requires association with several transcription coactivators for a successful cellular response to hypoxic stress. In addition to the conventional global transcription coactivator CREB-binding protein/p300 (CBP/p300) that binds to the HIF-α transactivation domain, a new group of transcription coactivators called the coiled-coil coactivators (CCCs) interact directly with the second PER-ARNT-SIM (PAS) domain of ARNT (ARNT PAS-B). These less studied transcription coactivators play essential roles in the HIF-dependent hypoxia response, and CCC misregulation is associated with several forms of cancer. To better understand CCC protein recruitment by the heterodimeric HIF transcription factor, we used x-ray crystallography, NMR spectroscopy, and biochemical methods to investigate the structure of the ARNT PAS-B domain in complex with the C-terminal fragment of a coiled-coil coactivator protein, transforming acidic coiled-coil coactivator 3 (TACC3). We found that the HIF-2α PAS-B domain also directly interacts with TACC3, motivating an NMR data-derived model suggesting a means by which TACC3 could form a ternary complex with HIF-2α PAS-B and ARNT PAS-B via β-sheet/coiled-coil interactions. These findings suggest that TACC3 could be recruited as a bridge to cooperatively mediate between the HIF-2α PAS-B·ARNT PAS-B complex, thereby participating more directly in HIF-dependent gene transcription than previously anticipated.


Concussion Risk and Recovery in Athletes With Psychostimulant-Treated Attention-Deficit/Hyperactivity Disorder: Findings From the NCAA-DOD CARE Consortium.
The aim of the present study was to examine the effects of attention-deficit/hyperactivity disorder (ADHD) -related psychostimulant use in the context of concussion risk and symptom recovery. Data were obtained from the National Collegiate Athletic Association Department of Defense Grand Alliance Concussion Assessment, Research, and Education (NCAA-DOD CARE) Consortium from 2014 to 2017. Relative to individuals without diagnosed ADHD (i.e., control), both ADHD diagnosis and the combination of ADHD diagnosis and psychostimulant use were associated with a greater risk of incurring a concussive injury. Following a concussive injury, ADHD diagnosis was associated with longer symptom recovery time relative to the control group. However, individuals with ADHD who use psychostimulants did not take longer to resolve symptoms than controls, suggesting that psychostimulants may have a positive influence on recovery. Regardless of time point, ADHD diagnosis was associated with an elevated number of concussion-related symptoms; however, this effect appears mitigated by having used ADHD-related psychostimulants.


Evaluating the performance of polygenic risk profiling across diverse ancestry populations in Parkinson's disease.
This study aims to address disparities in risk prediction by evaluating the performance of polygenic risk score (PRS) models using the 90 risk variants across 78 independent loci previously linked to Parkinson's disease (PD) risk across seven diverse ancestry populations. We conducted a multi-stage study, testing PRS models in predicting PD status across seven different ancestries applying three approaches: 1) PRS adjusted by gender and age; 2) PRS adjusted by gender, age and principal components (PCs); and 3) PRS adjusted by gender, age and percentage of population admixture. These models were built using the largest four population-specific summary statistics of PD risk to date (base data) and individual level data obtained from the Global Parkinson's Genetics Program (target data). We performed power calculations to estimate the minimum sample size required to conduct these analyses. A total of 91 PRS models were developed to investigate cumulative known genetic variation associated with PD risk and age of onset in a global context. We observed marked heterogeneity in risk estimates across non-European ancestries, including East Asians, Central Asians, Latino/Admixed Americans, Africans, African admixed, and Ashkenazi Jewish populations. Risk allele patterns for the 90 risk variants yielded significant differences in directionality, frequency, and magnitude of effect. PRS did not improve in performance when predicting disease status using similar base and target data across multiple ancestries, demonstrating that cumulative PRS models based on current known risk are inherently biased towards European populations. We found that PRS models adjusted by percentage of admixture outperformed models that adjusted for conventional PCs in highly admixed populations. Overall, the clinical utility of our models in individually predicting PD status is limited in concordance with the estimates observed in European populations. This study represents the first comprehensive assessment of how PRS models predict PD risk and age at onset in a multi-ancestry fashion. Given the heterogeneity and distinct genetic architecture of PD across different populations, our assessment emphasizes the need for larger and diverse study cohorts of individual-level target data and well-powered ancestry-specific summary statistics. Our current understanding of PD status unraveled through GWAS in European populations is not generally applicable to other ancestries. Future studies should integrate clinical and *omics level data to enhance the accuracy and predictive power of PRS across diverse populations.


Toxicant Responses and Culturing Characteristics of Long-Term Laboratory-Reared and Field Populations of Ceriodaphnia dubia.
Ceriodaphnia dubia is a standardized test organism for regulatory toxicity testing of surface waters and commercial chemicals because of its simplicity to culture and responsiveness to toxicants. For testing convenience, C. dubia is often cultured for extended periods in the laboratory with little knowledge of the impact on subsequent generations. Extended laboratory rearing could impact how they respond to stressors and decrease the accuracy of test results. The present study investigated if C. dubia cultured for an extended period were representative of three recently collected field populations by comparing their culturing characteristics and sensitivities to toxicants. For culturing characteristics, the field cultures were more challenging because they had shorter body lengths, fewer neonates, and higher mortality rates than the laboratory culture. Comparative chronic toxicity tests with sodium chloride and the neonicotinoid insecticide thiamethoxam indicated that the laboratory and field organisms did not differ much in their toxicological responses but did differ in the variability of responses (percentage of coefficient of variation). The differences between the laboratory and field cultures found in the present study highlight the challenges of addressing discrepancies between laboratory and field applications in existing standardized methodologies. Environ Toxicol Chem 2023;00:1-11. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.


[Fatigue in patients following traumatic brain injury and stroke].
Fatigue may be defined as an experience of long-term or recurrent tiredness and reduced capacity for mental and/or physical activity, and is common following acquired brain injury. This article provides an overview of the status of knowledge on fatigue following stroke and traumatic brain injury. An assessment of the existing literature, including a literature search, as well as the authors' own clinical experience form the basis for the content of this article. Existing studies use varying definitions of fatigue and apply different survey tools. The prevalence of fatigue following stroke or traumatic brain injury is uncertain due to varying inclusion criteria and survey methods, and because different times of measurement can result in different prevalence figures. Several studies estimate the prevalence to be between 50 and 70%. Our knowledge of the pathologic mechanisms is very limited; premorbid, sociodemographic, stroke/trauma-related and comorbid factors all play a role in the experience of fatigue. Therapeutic interventions include psychoeducation, regulation of activity, physical exercise and treatment of comorbid conditions. Trials have also been conducted on drug treatment, so far without convincing results. Fatigue occurs frequently in stroke patients and patients with traumatic brain injuries, and often significantly restricts daily functioning for these patients. Our knowledge of pathologic mechanisms and evidence with regard to therapeutic interventions is deficient, and the patient population is heterogeneous. The treatment is adapted to the individual and its objective is to enable individuals to use their limited capacity in the most appropriate way possible.


Impact of anti-tacking agents on properties of gas-entrapped membrane and effervescent floating tablets.
Tackiness caused by the gas-entrapped membrane (Eudragit(®)RL 30D) was usually observed during storage of the effervescent floating tablets, leading to failure in floatation and sustained release. In this work, common anti-tacking agents (glyceryl monostearate (GMS) and talc) were used to solve this tackiness problem. The impact of anti-tacking agent on the properties of free films and corresponding floating tablets was investigated. GMS was more effective than talc in reducing tackiness of the film. Addition and increasing amount of anti-tacking agents lowered the film mechanical strength, but the coating films were still strong and flexible enough to resist the generated gas pressure inside the floating tablet. Wettability and water vapor permeability of the film decreased with increasing level of anti-tacking agents as a result of their hydrophobicity. No interaction between anti-tacking agents and polymer was observed as confirmed by Fourier transform infrared spectroscopy, powder X-ray diffractometry, and differential scanning calorimetry studies. Increasing amount of anti-tacking agents decreased time to float and tended to retard drug release of the floating tablets. Floating properties and drug release were also influenced by type of anti-tacking agents. The obtained floating tablets still possessed good floating properties and controlled drug release even though anti-tacking agent had some effects. The results demonstrated that the tackiness problem of the floating tablets could be solved by incorporating anti-tacking agent into the gas-entrapped membrane.


Site-selective incorporation of phosphorylated tyrosine into the p50 subunit of NF-κB and activation of its downstream gene CD40.
The NF-κB family of transcriptional activators is responsible for the expression of numerous genes that control key functions such as cell development and survival. Subunit p50 has been studied extensively and is known to include 13 tyrosines, but the extent and pattern of tyrosine phosphorylation that accompanies p50 function has not been defined in the literature, especially at the level of selectivity of gene expression. In this study, phosphorylated tyrosine (pTyr) was site-selectively incorporated into the p50 subunit using an E. coli in vitro expression system containing a modified ribosome. In human T cells, the NF-κBs containing a pTyr at position 60 or 82 of p50 strongly increased the expression of CD40, which is a potential target for cancer or viral immunotherapy. Promoter DNA binding was studied for CD40 promoters, and verified two pTyr residues in NF-κB p50/p65 heterodimers that facilitated this process, and that support the possible importance of phosphorylation stoichiometry. This study defines a new approach for studying tyrosine residues whose phosphorylation alters protein binding to DNA promoters, and contributes to the facility of DNA expression.


The SigmaR1 chaperone drives breast and colorectal cancer cell migration by tuning SK3-dependent Ca2+ homeostasis.
The remodeling of calcium homeostasis contributes to the cancer hallmarks and the molecular mechanisms involved in calcium channel regulation in tumors remain to be characterized. Here, we report that SigmaR1, a stress-activated chaperone, is required to increase calcium influx by triggering the coupling between SK3, a Ca2+-activated K+ channel (KCNN3) and the voltage-independent calcium channel Orai1. We show that SigmaR1 physically binds SK3 in BC cells. Inhibition of SigmaR1 activity, either by molecular silencing or by the use of sigma ligand (igmesine), decreased SK3 current and Ca2+ entry in breast cancer (BC) and colorectal cancer (CRC) cells. Interestingly, SigmaR1 inhibition diminished SK3 and/or Orai1 levels in lipid nanodomains isolated from BC cells. Analyses of tissue microarray from CRC patients showed higher SigmaR1 expression levels in cancer samples and a correlation with tumor grade. Moreover, the exploration of a cohort of 4937 BC patients indicated that high expression of SigmaR1 and Orai1 channels was significantly correlated to a lower overall survival. As the SK3/Orai1 tandem drives invasive process in CRC and bone metastasis progression in BC, our results may inaugurate innovative therapeutic approaches targeting SigmaR1 to control the remodeling of Ca2+ homeostasis in epithelial cancers.


A Validated Nomogram to Predict Upstaging of Ductal Carcinoma in Situ to Invasive Disease.
Approximately 8-56% of patients with a core needle biopsy (CNB) diagnosis of ductal carcinoma in situ (DCIS) will be upstaged to invasive disease at the time of excision. Patients with invasive disease are recommended to undergo axillary nodal staging, most often requiring a second operation. We developed and validated a nomogram to preoperatively predict percentage of risk for upstaging to invasive cancer. We reviewed 834 cases of DCIS on CNB between January 2004 and October 2014. Multivariable analysis was used to evaluate CNB and imaging factors to develop a nomogram to predict the risk of upstaging from DCIS to invasive cancer. This nomogram was validated with an external dataset of 579 similar patients between November 1998 and September 2016. An area under the receiver operating characteristic curve was constructed to evaluate nomogram discrimination. The rate of upstaging to invasive disease was 118/834 (14.1%). On multivariable analysis, grade on CNB and imaging factors, including mass lesion, multicentric disease, and largest linear dimension, were associated with upstage to invasive disease, and was used to develop a nomogram (c-statistic 0.71). In the external validation dataset, 62/579 (10.7%) patients were upstaged to invasive disease. Our nomogram was validated in this dataset with a c-statistic of 0.71. For patients with a CNB diagnosis of DCIS, our validated nomogram using DCIS grade on biopsy, and imaging factors of mass lesion, multicentric disease, and largest linear dimension, may be used for preoperative assessment of risk of upstaging to invasive disease, allowing patient counseling regarding axillary staging at the time of definitive surgery.


Molecular and clinical characteristics of MSH6 germline variants detected in colorectal cancer patients.
The MSH6 gene is one of the mismatch repair genes involved in Lynch syndrome and its mutations account for 10-20% of Lynch syndrome. Although previous studies suggested that the difference of the geographical region affects the clinical phenotype of Lynch syndrome, there has been no report on the detailed features of Japanese Lynch syndrome patients carrying an MSH6 mutation. The aim of the present study was to investigate the clinical and molecular features of MSH6 mutation carriers in Japan. Surgically resected 1720 colorectal carcinoma specimens were screened by microsatellite instability (MSI) testing and the MSI-high cases were subjected to a germline mutation analysis of the mismatch repair genes MLH1, MSH2 and MSH6. We investigated the clinical and molecular features of the MSH6 variants, such as the family cancer history, pathological findings, immunohistochemistry, methylation status of the MLH1 promoter and BRAF mutation in the colorectal tumor. Furthermore, the impact of the missense variants on MSH6 protein was predicted by using in silico tools. We identified nine novel pathogenic mutations and eight unclassified missense variants. Among the eight missense variants, three were suspected pathogenic by in silico analysis. We also found that most colorectal cancers in the MSH6 mutation carrier were diagnosed after the age of 50 and were localized distally. Furthermore, the mean age at diagnosis of endometrial cancer in Japanese MSH6 mutation carriers (49.2 years) was earlier than previous reports from Western countries (56.5 years). These results may improve the surveillance program for Japanese MSH6 mutation carriers.


Evaluation of autologous bone marrow mesenchymal stem cell-calcium phosphate ceramic composite for lumbar fusion in rhesus monkey interbody fusion model.
Autologous bone marrow mesenchymal stem cell (BMSC)-calcium phosphate ceramic composites were constructed in vitro and implanted as a bone graft substitute for lumbar anterior interbody fusion in rhesus monkeys to determine the osteogenic capacity of the composites. Nine adult rhesus monkeys underwent lumbar L3-L4 and L5-L6 diskectomy and interbody fusion via an anterior retroperitoneal approach. Two fusion sites in each animal were randomly assigned to two of three treatments: autogenous tricortical iliac crest bone graft (autograft group), cell-free ceramic graft (ceramic group), or BMSC-ceramic composite graft (BMSC group). Autologous BMSCs were expanded in culture and stimulated with osteogenic supplement. The spinal fusion segments were evaluated by radiography, biomechanical testing, histologic analysis, and histomorphometric analysis 3 months postsurgery. The BMSC group achieved lumbar interbody fusion superior to that of the ceramic group, both biomechanically and histologically. The BMSC group and the autograft group showed equivalent biomechanical stiffness. Ceramic residues were significantly greater in the ceramic group versus the BMSC group. The results indicate that BMSC-ceramic composites can enhance bone regeneration and achieve osseous spinal fusion 3 months after implantation in the rhesus monkey interbody fusion model.


Evaluation of a polyclonal antibody for the detection and identification of ranaviruses from freshwater fish and amphibians.
A rabbit polyclonal antibody (PAb) raised against European catfish virus (ECV; isolated from black bullhead Ameiurus melas in France) was produced and then evaluated using a panel of 9 ranavirus isolates collected from different lower vertebrate species originating from Australia, North and South America, Southeast Asia, and Europe. Using ranavirus-infected epithelioma papillosum cyprini (EPC) cell cultures, the specificity of the PAb was determined by Western blot, immunogold electron microscopy, and direct enzyme-linked immunosorbent assay (ELISA). Western blot analysis demonstrated that the PAb reacted strongly with a protein with a molecular weight corresponding to approximately 49 kDa. Immunogold electron microscopy provided direct evidence that the epitopes recognized by this PAb were located on the outer surface of virions. The PAb was used for the preparation of a peroxidase-labeled conjugate for the direct ELISA detection of ranaviruses in infected EPC cell cultures. The specificity of the conjugated PAb was tested using ranaviruses, some representative fish viruses of the genera Rhabdovirus and Birnavirus, and samples from various non-infected fish species. The PAb detected all tested ranaviruses except for 2 Santee-Cooper ranaviruses. The direct ELISA enabled the detection of ranavirus from a concentration of 10(3.5) to 10(3.8) TCID50 ml(-1) cell culture. The results of this study revealed that the rabbit PAb raised against ECV could be useful for the development of specific and standardized diagnostic assays for the detection of ranaviruses from freshwater fish and amphibians.


Relationship between sonographic parameters and YKL-40 levels in rheumatoid arthritis.
YKL-40, also known as human cartilage glycoprotein 39, is a member of the "mammalian chitinase-like proteins" family without chitinase activity. Increased serum concentrations are associated with inflammatory processes and several types of cancer. In this study, we evaluated YKL-40 levels in serum and synovial fluid of patients with rheumatoid arthritis in comparison with the ultrasonographic findings. YKL-40 levels were measured by enzyme-linked immunosorbent assay in 25 patients with active rheumatoid arthritis and in 40 healthy subjects. B mode and power Doppler were performed to determine synovial thickening and vascularization. Serum YKL-40 level in patients was significantly higher than the concentration in healthy controls (P < 0.01). In patients with rheumatoid arthritis, the level of the glycoprotein in synovial fluid was remarkably elevated compared to the serum level (P = 0.003). The B mode and power Doppler scores correlated to YKL-40 in serum and synovial fluid (P = 0.07). Serum YKL-40 levels were related positively to serum markers of inflammation such as C-reactive protein (P = 0.004) and erythrocyte sedimentation rate (P = 0.003). This study is the first to demonstrate a relationship between YKL-40 levels and ultrasonographic examinations in Bulgarian patients with rheumatoid arthritis. The findings suggest that YKL-40 might be implicated in the pathogenesis of the disease and could indicate the level of joint inflammation.


Purification and characterization of a soybean flour-inducible ferredoxin reductase of Streptomyces griseus.
We have purified an NADH-dependent ferredoxin reductase from crude extracts of Streptomyces griseus cells grown in soybean flour-enriched medium. The purified protein has a molecular weight of 60,000 as determined by sodium dodecyl sulfate gel electrophoresis. The enzyme requires Mg2+ ion for catalytic activity in reconstituted assays, and its spectral properties resemble those of many other flavin adenine dinucleotide-containing flavoproteins. A relatively large number of hydrophobic amino acid residues are found by amino acid analysis, and beginning with residue 7, a consensus flavin adenine dinucleotide binding sequence, GXGXXGXXXA, is revealed in this protein. In the presence of NADH, the ferredoxin reductase reduces various electron acceptors such as cytochrome c, potassium ferricyanide, dichlorophenolindophenol, and nitroblue tetrazolium. However, only cytochrome c reduction by the ferredoxin reductase is enhanced by the addition of ferredoxin. In the presence of NADH, S. griseus ferredoxin and cytochrome P-450soy, the ferredoxin reductase mediates O dealkylation of 7-ethoxycoumarin.


Evolutionary Origin of the Staphylococcal Cassette Chromosome mec (SCCmec).
Several lines of evidence indicate that the most primitive staphylococcal species, those of the Staphylococcus sciuri group, were involved in the first stages of evolution of the staphylococcal cassette chromosome mec (SCCmec), the genetic element carrying the β-lactam resistance gene mecA However, many steps are still missing from this evolutionary history. In particular, it is not known how mecA was incorporated into the mobile element SCC prior to dissemination among Staphylococcus aureus and other pathogenic staphylococcal species. To gain insights into the possible contribution of several species of the Staphylococcus sciuri group to the assembly of SCCmec, we sequenced the genomes of 106 isolates, comprising S. sciuri (n = 76), Staphylococcus vitulinus (n = 18), and Staphylococcus fleurettii (n = 12) from animal and human sources, and characterized the native location of mecA and the SCC insertion site by using a variety of comparative genomic approaches. Moreover, we performed a single nucleotide polymorphism (SNP) analysis of the genomes in order to understand SCCmec evolution in relation to phylogeny. We found that each of three species of the S. sciuri group contributed to the evolution of SCCmec: S. vitulinus and S. fleurettii contributed to the assembly of the mec complex, and S. sciuri most likely provided the mobile element in which mecA was later incorporated. We hypothesize that an ancestral SCCmec III cassette (an element carried by one of the most epidemic methicillin-resistant S. aureus clones) originated in S. sciuri possibly by a recombination event in a human host or a human-created environment and later was transferred to S. aureus.


Coating membranes for a sorbent-based artificial liver: adsorption characteristics.
Techniques are described for the coating of sorbents to be used in an artificial liver support system based on mixed sorbent bed hemoperfusion. Activated charcoal has been coated with cellulose acetate (CA) by solvent evaporation. With Amberlite XAD-4, the Wurster technique was used for coating with CA. XAD-4 has also been coated with a synthetic polyelectrolyte with anticoagulant activity by adsorption and fixation by gamma radiation-induced crosslinking. Activated charcoal, XAD-4, and a cation exchange resin, all in powdered form, were encapsulated in agarose gel beads. Adsorption characteristics onto the sorbents are described. The results are in agreement with a theoretical model presented. In general, adsorption onto XAD-4 is limited by film diffusion. With activated charcoal, pore diffusion limitation is generally observed. Blood compatibility is improved by coating.


A five century evolution of cervical incompetence as a clinical entity.
Since cervical incompetence was introduced in the English literature in 1678, our understanding and obstetric management of this clinical entity, have changed tremendously over the years. This review shows the historical perspective of the development of cervical incompetence as a distinct clinical entity and an all or nothing phenomenon to cervical incompetence as part of a spectrum leading to preterm delivery, which can express differently in subsequent pregnancies. These changes in our understanding imply consequences for the obstetric management of cervical incompetence. This review focuses on the obstetric management of women considered to be at high risk of preterm delivery due to cervical incompetence, by transvaginal ultrasonographic follow-up of cervical length and transvaginal cervical cerclage.


Evaluation of cellular immunologic responsiveness in the clinical management of patients with prostatic cancer. II. Effect of oestrogen, cryosurgery and transurethral resection on thymic-dependent lymphocytic blastogenesis.
The effect of oestrogen, cryosurgery and transurethral resection (TUR) of the prostate on the blastogenic response of thymic-dependent peripheral blood lymphocytes (PBL) to the non-specific mitogen, phytohaemagglutinin (PHA) was evaluated as one in vitro criteria of each of these treatment modalities on the cellular immunologic responsiveness of 24 patients with prostatic cancer. A depression 5 days following receipt of oestrogen and 2-7 days following cryosurgery or TUR of the responsiveness of PHA-stimulated PBL was observed. Oestrogen-induced aberrations of responsiveness may not only be of relevance in prostatic cancer patients, but also suggested association between uterine cancer and prolonged administration of diethylstilboesterol and the development of vaginal tumours in offspring found in association with maternal ingestion during pregnancy. Particularly striking was that contrary to the reduced responsiveness of PBL cultured in autologous and homologous serum from patients receiving TUR, patients receiving cryosurgery, while also showing reduction in autologous serum, showed increased responsiveness when cultured in homologous serum. Although transient, depression of lymphocyte responsiveness, particularly if involving tumour-cloned T cells, may provide reduced surveillance to potential tumour cells leading to an alteration of tumour-host homeostasis. The potential of reduced tumour surveillance at least in the case of TUR, appears to be supported by observations that patients expiring from prostatic cancer at our institution had an antecedent TUR. The possibility of identifying those patients possessing aberrations of responsiveness prior to therapy, as well as those prone to develop or undergo further reduction in their responsiveness following the presently evaluated treatment modalities would appear to be of real and relevant concern in the management of the patient with prostatic, as well as other types of malignant neoplasms. The possibility of pre-operative and/or post-operative immunotherapy in such patients may be indicated pending further study.


Multiple receptor tyrosine kinases are expressed in adult rat retinal ganglion cells as revealed by single-cell degenerate primer polymerase chain reaction.
To achieve a better understanding of the repertoire of receptor tyrosine kinases (RTKs) in adult retinal ganglion cells (RGCs) we performed polymerase chain reaction (PCR), using degenerate primers directed towards conserved sequences in the tyrosine kinase domain, on cDNA from isolated single RGCs univocally identified by retrograde tracing from the superior colliculi. All the PCR-amplified fragments of the expected sizes were sequenced, and 25% of them contained a tyrosine kinase domain. These were: Axl, Csf-1R, Eph A4, Pdgfrbeta, Ptk7, Ret, Ros, Sky, TrkB, TrkC, Vegfr-2, and Vegfr-3. Non-RTK sequences were Jak1 and 2. Retinal expression of Axl, Csf-1R, Pdgfrbeta, Ret, Sky, TrkB, TrkC, Vegfr-2, and Vegfr-3, as well as Jak1 and 2, was confirmed by PCR on total retina cDNA. Immunodetection of Csf-1R, Pdgfralpha/beta, Ret, Sky, TrkB, and Vegfr-2 on retrogradely traced retinas demonstrated that they were expressed by RGCs. Co-localization of Vegfr-2 and Csf-1R, of Vegfr-2 and TrkB, and of Csf-1R and Ret in retrogradely labelled RGCs was shown. The effect of optic nerve transection on the mRNA level of Pdgfrbeta, Csf-1R, Vegfr-2, Sky, and Axl, and of the Axl ligands Gas6 and ProteinS, was analysed. These analyses show transection-induced changes in Axl and ProteinS mRNA levels. The repertoire of RTKs expressed by RGCs is more extensive than previously anticipated. Several of the receptors found in this study, including Pdgfrbeta, Csf-1R, Vegfr-2, Sky, and Axl, and their ligands, have not previously been primarily associated with retinal ganglion cells.


A relative ordering-based predictor for tamoxifen-treated estrogen receptor-positive breast cancer patients: multi-laboratory cohort validation.
Current predictors for estrogen receptor-positive (ER-positive) breast cancer patients receiving tamoxifen are often invalid in inter-laboratory validation. We aim to develop a robust predictor based on the relative ordering of expression measurement (ROE) in gene pairs. Using a large integrated dataset of 420 normal controls and 1,129 ER-positive breast tumor samples, we identified the gene pairs with stable ROEs in normal control and significantly reversed ROEs in ER-positive tumor. Using these gene pairs, we characterized each sample of a cohort of 292 ER-positive patients who received tamoxifen monotherapy for 5 years and then identified relapse risk-associated gene pairs. We extracted a gene pair subset that resulted in the largest positive and negative predictive values for predicting 10-year relapse-free survival (RFS) using a genetic algorithm. A predictor was developed based on the gene pair subset and was validated in 2 large multi-laboratory cohorts (N = 250 and 248, respectively) of ER-positive patients who received 5-year tamoxifen alone. In the first validation cohort, the patients predicted to be tamoxifen sensitive had a 10-year RFS of 91 % (95 % confidence interval [CI] 85-97 %) with an absolute risk reduction of 34 % (95 % CI 17-51 %). The patients predicted to be tamoxifen insensitive had a significantly higher relapse risk than the patients predicted to be tamoxifen sensitive (hazard ratio = 4.99, 95 % CI 2.45-10.17, P = 9.13 × 10(-7)). Similar performance was achieved for the second validation cohort. The predictor performed well in both node-negative and node-positive subsets and added significant predictive power to the clinical parameters. In contrast, 2 previously proposed predictors did not achieve significantly better performances than the baselines of the validation cohorts. In summary, the proposed predictor can accurately and robustly predict tamoxifen sensitivity of ER-positive breast cancer patients and identified patients with a high probability of 10-year RFS following tamoxifen monotherapy.


Mid-sized groups perform best in a collective decision task in sticklebacks.
Numerous studies have reported functional improvements in collective behaviour with increasing group size, however, the possibility that such improvements may saturate or even decline as group size continues to grow have seldom been tested experimentally. Here, we tested the ability of solitary three-spined sticklebacks and those in groups, ranging from 2 to 29 fish, to leave an unfavourable patch of habitat. Our results replicate the findings of previous studies at low group sizes, with the fish initially showing a reduction in their latency to leave the unfavourable habitat as group size increased. As group size continued to increase, however, latency to leave the habitat increased, so that the functional relationship between group size and latency to depart was U-shaped. Our results suggest an optimum group size in this context of between 12 and 20 fish. Underlying this group-level trend was a similar U-shaped relationship between group size and the first fish to leave the habitat, suggesting that at larger group sizes, social conformity to the behaviour of the majority can stifle the ability of fish to innovate-in this case, to induce a collective movement from the unfavourable habitat.


Self-esteem and oral health behavior in adolescents.
This study investigated the mediation of self-esteem in adolescents' oral health behaviors. The Rosenberg self-esteem scale was used to assess self-esteem, whereas data from socio-demographic and behavior characteristics were analyzed by questions validated in previous surveys. The teenagers had good oral health behavior, except unhealthy diet. The number of adolescents with high self-esteem was a lot smaller than those with low self-esteem. The use of dental services, even when associated with high self-esteem, lost significance after being adjusted by sex, age and tooth brushing frequency. Nevertheless, multiple logistic regression analysis, using unadjusted estimates and adjusted with their respective Confidence Intervals of 95%, showed a relationship of self-esteem with age (p-value=0.001) and tooth brushing frequency (p-value=0.019). Regardless of the sex, students over 16 years old with high self-esteem brush their teeth more often, having probably better oral health. These results confirm the modulation of self-esteem in oral health, and then it is necessary the analysis and the use of these psychosocial factors in the young oral health care.


[Natural foci of tick-borne encephalitis on the southwestern coast of Sakhalin].
Nucleic acids of Ixodes persulcatus were studied by molecular hybridization in the natural focus of tick-borne encephalitis in Kholmsk District of Sakhalin Province. The studies have shown wide dissemination of viral RNA in the focus. The infectivity of ticks in various sites of habitation varied from 3.5 to 18.5%, their number fluctuating from 0.4 to 300 and more imago per flag-hour. The most active part of the natural focus has been determined using zoological-viral indexes. The viral strain of tick-borne encephalitis has been isolated.


Resolving multiphoton processes with high-order anisotropy ultrafast X-ray scattering.
We present the first results on experimentally measured ultrafast X-ray scattering of strongly driven molecular iodine and analysis of high-order anisotropic components of the scattering signal. We discuss the technical details of retrieving high fidelity high-order anisotropy components from the measured scattering data and outline a method to analyze such signals using Legendre decomposition. We describe how anisotropic motions can be extracted from the various Legendre orders using simulated anisotropic scattering signals and Fourier analysis. We implement the method on the measured signal and observe a multitude of dissociation and vibration motions simultaneously arising from various multiphoton transitions occurring in the sample. We use the anisotropic scattering information to disentangle the different processes and assign their dissociation velocities on the Angstrom and femtosecond scales de novo.


Investigating Genetic Mutations in a Large Cohort of Iranian Patients with Congenital Hyperinsulinism
Congenital hyperinsulinism (CHI) is the most frequent cause of severe and persistent hypoglycaemia from birth. Understanding the pathophysiology and genetic defects behind hyperinsulinism and its complications provides clues to timely diagnosis and management. The aim of this study was to evaluate the underlying genetic aetiology of a specific Iranian pediatric cohort with CHI. A total of 44 unrelated children, 20 girls and 24 boys, with an initial diagnosis or history of CHI from all regions of Iran were recruited between 2016 and 2019. Targeted next generation sequencing (tNGS) was performed for the genes found in about half of CHI patients. Mutations were identified in 24 cases (55%). Patients with a confirmed genetic cause were mainly diagnosed below age of one year old (p=0.01), had fewer other syndromic features, excluding seizure, (p=0.03), were less diazoxide responsive (p=0.04) and were more diazoxide unresponsive leading to pancreatectomy (p=0.007) compared to those with no identified mutations. Among 24 patients with identified genetic mutations, 17 (71%) had a mutation in ABCC8, 3 (12%) in KCNJ11, 3 (12%) in HADH, and 1 patient had a mutation in KMT2D. These included five novel mutations in ABCC8, KCNJ11, and KMT2D. This is the biggest genetic study of CHI in Iran. A high frequency of recessive forms of CHI, especially HADH mutations, in our study could be due to a high rate of consanguineous marriage. We recommend tNGS to screen for all the CHI genes.


The FHIT gene product: tumor suppressor and genome "caretaker".
The FHIT gene at FRA3B is one of the earliest and most frequently altered genes in the majority of human cancers. It was recently discovered that the FHIT gene is not the most fragile locus in epithelial cells, the cell of origin for most Fhit-negative cancers, eroding support for past claims that deletions at this locus are simply passenger events that are carried along in expanding cancer clones, due to extreme vulnerability to DNA damage rather than to loss of FHIT function. Indeed, recent reports have reconfirmed FHIT as a tumor suppressor gene with roles in apoptosis and prevention of the epithelial-mesenchymal transition. Other recent works have identified a novel role for the FHIT gene product, Fhit, as a genome "caretaker." Loss of this caretaker function leads to nucleotide imbalance, spontaneous replication stress, and DNA breaks. Because Fhit loss-induced DNA damage is "checkpoint blind," cells accumulate further DNA damage during subsequent cell cycles, accruing global genome instability that could facilitate oncogenic mutation acquisition and expedite clonal expansion. Loss of Fhit activity therefore induces a mutator phenotype. Evidence for FHIT as a mutator gene is discussed in light of these recent investigations of Fhit loss and subsequent genome instability.


Aberrant intervertebral motion in patients with treatment-resistant nonspecific low back pain: a retrospective cohort study and control comparison.
Intervertebral kinematic assessments have been used to investigate mechanical causes when back pain is resistant to treatment, and recent studies have identified intervertebral motion markers that discriminate patients from controls. However, such patients are a heterogeneous group, some of whom have structural disruption, but the effects of this on intervertebral kinematics are unknown. Thirty-seven patients with treatment-resistant back pain referred for quantitative fluoroscopy were matched to an equal number of pain-free controls for age and sex. All received passive recumbent flexion assessments for intervertebral motion sharing inequality (MSI), variability (MSV), laxity and translation. Comparisons were made between patient subgroups, between patients and controls and against normative levels from a separate group of controls. Eleven patients had had surgical or interventional procedures, and ten had spondylolisthesis or pars defects. Sixteen had no disruption. Patients had significantly higher median MSI values (0.30) than controls (0.27, p = 0.010), but not MSV (patients 0.08 vs controls 0.08, p = 0.791). Patients who received invasive procedures had higher median MSI values (0.37) than those with bony defects (0.30, p = 0.018) or no disruption (0.28, p = 0.0007). Laxity and translation above reference limits were not more prevalent in patients. Patients with treatment-resistant nonspecific back pain have greater MSI values than controls, especially if the former have received spinal surgery. However, excessive laxity, translation and MSV are not more prevalent in these patients. Thus, MSI should be investigated as a pain mechanism and for its possible value as a prognostic factor and/or target for treatment in larger patient populations. These slides can be retrieved under Electronic Supplementary Material.


[Choice of surgical approach in the treatment of patients with deep frost bites].
Two main approaches to surgical treatment were studied in 114 patients with deep frost bites of the extremities. First - early necrectomy with subsequent local treatment of wounds and their surgical closure on the final stage. Second - long-term conservative treatment until formation of demarcation line and limitation of necrotic tissues with subsequent surgical treatment including their resection and simultaneous formation of the stump of the extremity. It is demonstrated that active surgical policy has significant advantages over expectant one as evident from reduce time of treatment and less number of postoperative pyonecrotic complications (21,7% vs 78,4%).


[Giant mesenchymal tumor of the vulva with atypical clinical evolution].
Mixed mesenchymal tumors of the vulva are rare lesions. On other hand, aggressive angiomyxoma is a recently characterized neoplasm occurring principally in the pelvic soft-tissues and showing propensity for local recurrence. The benign soft-tissue tumors are small than malign neoplasm, mimicking and often are misdiagnosed as cysts of Bartholin's gland. In this report we describe one case of a benign, giant and atypical mixed mesenchymal tumor of the vulva and its particular clinical characteristics.


Surface-enhanced Raman scattering from transition metals with special surface morphology and nanoparticle shape.
This discussion focuses on our recent approaches at aiming to optimize surface-enhanced Raman scattering (SERS) activity for transition metals (group VIII B elements), by intentionally fabricating desired surface nanostructures or synthesizing nanoparticles. The SERS activity of transition metals critically depends on the surface morphology of electrodes and on size, shape and aggregation form of nanoparticles. A correct surface roughening procedure for transition-metal electrodes is indispensable for fabricating cauliflower-like nanostructures that show a higher SERS activity. Two more methods have been explored to synthesize nanoparticles, i.e., cubic nanoparticles and gold-core palladium-shell nanostructures, respectively. Their SERS activities are considerably higher than those of normal spherical mono-metallic nanoparticles. To explain these observations, a preliminary theoretical calculation, using the three-dimensional finite difference time domain (3D-FDTD) method, was performed to evaluate the local electromagnetic field on transition metal surfaces. The result is in good agreement with the experimental data.


Association of stressful life events and psychological problems profile: Results from a large-scale cross-sectional study among Iranian industrial employees using Bayesian quantile structural equation model.
Objectives: The current study aimed at evaluating the major domains of stressful life events and their association with psychological problems profile in a large sample of Iranian industrial manufacturing employees. Methods: In a cross-sectional study, 3,063 participants were randomly selected from 16,000 employees working in a big industrial company in Isfahan, Iran. Three common psychological problems i.e. depression, anxiety and psychological distress were evaluated using Persian validated version of Hospital Anxiety and Depression Scale (HADS) and 12-item General Health Questionnaire (GHQ-12), respectively. Self-perceived frequency and intensity of stressful life events were measured by stressful life event (SLE) questionnaire. Bayesian quantile structural equation model in R free statistical software (version 3.4) was used for evaluating the association of stressful life events and levels of psychological problems profile. Results: Using factor analysis, two major domains i.e. socioeconomic and personal stressors were derived from 11 life stressors and a unified measure i.e. psychological problem profile was extracted from three common psychological problems. Financial and daily life stressors had the highest and sexual problems showed lowest intensity. Quantile structural equation model revealed that the psychological problems profile scores had stronger association with personal (β: ranging from 0.45, 1.87) than socioeconomics stressors (β: ranging from 0.11, 0.27). The association of socioeconomic stressors was fairly uniform across quantiles of psychological problems scores, while personal stressors showed stronger association in higher quantiles, meaning that employees with higher mental health problems more experienced personal stressors. Conclusions: Life stressors particularly personal showed direct association with intensity of psychological problems in manufacturing employees. Life stressors are more perceived by employees with higher intensity of psychological problems. The results of current study can be useful in planning occupational health programs in order to improve psychological health and job productivity.


Dietary pattern of self-care among Asian and Caucasian diabetic patients.
Diabetes mellitus is a growing pandemic and its self-care management rests primarily with the individual. This qualitative case study investigated the self-care dietary pattern among a group of 25 Asians and 24 Caucasians diagnosed with type 1 or type 2 diabetes. Data collected from a semi-structured interview and a 7-day health diary explored the self-care activities undertaken by the participants to establish metabolic control. From an analytical perspective, the collective responses were placed on a continuum ranging from strict adherence, moderately flexible adherence to very flexible adherence. The findings suggest that most of the participants were located in the latter two categories of diet related to self-care. Implications for healthcare professionals in promoting self-care will be discussed.


New and existing pharmacotherapeutic options for persistent asthma and COPD.
Asthma and COPD are chronic inflammatory airway disorders with systemic manifestations. The two diseases have different airway inflammation, features of airway remodelling with subsequent pathophysiology and clinical presentation. The international management guidelines recommend stepwise pharmacotherapy depending on disease control and÷or disease stage, comprising relievers and overall uniform controller treatment, despite the heterogeneity across the conditions and treatment response. Despite effective medications per se, still too many patients remain uncontrolled and no treatment can definitely cure either of the conditions. This overview includes currently recommended pharmacotherapeutic options with novel and future treatment targets.


Unraveling the mechanisms responsible for the comorbidity between metabolic syndrome and mental health disorders.
The increased prevalence and high comorbidity of metabolic syndrome (MetS) and mental health disorders (MHDs) have prompted investigation into the potential contributing mechanisms. There is a bidirectional association between MetS and MHDs including schizophrenia, bipolar disorder, depression, anxiety, attention-deficit/hyperactivity disorder, and autism spectrum disorders. Medication side effects and social repercussions are contributing environmental factors, but there are a number of shared underlying neurological and physiological mechanisms that explain the high comorbidity between these two disorders. Inflammation is a state shared by both disorders, and it contributes to disruptions of neuroregulatory systems (including the serotonergic, dopaminergic, and neuropeptide Y systems) as well as dysregulation of the hypothalamic-pituitary-adrenal axis. MetS in pregnant women also exposes the developing fetal brain to inflammatory factors that predispose the offspring to MetS and psychopathologies. Due to the shared nature of these conditions, treatment should address aspects of both mental health and metabolic disorders. Additionally, interventions that can interrupt the transfer of increased risk of the disorders to the next generation need to be developed. © 2013 S. Karger AG, Basel.


Frailty and risk of falls, fracture, and mortality in older women: the study of osteoporotic fractures.
A standard phenotype of frailty was associated with an increased risk of adverse outcomes including mortality in a recent study of older adults. However, the predictive validity of this phenotype for fracture outcomes and across risk subgroups is uncertain. To determine whether a standard frailty phenotype was independently associated with risk of adverse health outcomes in older women and to evaluate the consistency of associations across risk subgroups defined by age and body mass index (BMI), we ascertained frailty status in a cohort of 6724 women>or=69 years and followed them prospectively for incident falls, fractures, and mortality. Frailty was defined by the presence of three or more of the following criteria: unintentional weight loss, weakness, self-reported poor energy, slow walking speed, and low physical activity. Incident recurrent falls were defined as at least two falls during the subsequent year. Incident fractures (confirmed with x-ray reports), including hip fractures, and deaths were ascertained during an average of 9 years of follow-up. After controlling for multiple confounders such as age, health status, medical conditions, functional status, depressive symptoms, cognitive function, and bone mineral density, frail women were subsequently at increased risk of recurrent falls (multivariate odds ratio=1.38, 95% confidence interval [CI], 1.02-1.88), hip fracture (multivariate hazards ratio [MHR]=1.40, 95% CI, 1.03-1.90), any nonspine fracture (MHR=1.25, 95% CI, 1.05-1.49), and death (MHR=1.82, 95% CI, 1.56-2.13). The associations between frailty and these outcomes persisted among women>or=80 years. In addition, associations between frailty and an increased risk of falls, fracture, and mortality were consistently observed across categories of BMI, including BMI>or=30 kg/m2. Frailty is an independent predictor of adverse health outcomes in older women, including very elderly women and older obese women.


Sexual Media and Childhood Well-being and Health.
Sexual content is highly prevalent in traditional media, and portrayals rarely depict the responsibilities and risks (eg, condom use, pregnancy) associated with sexual activity. Exposure to such content is linked with shifts in attitudes about sex and gender, earlier progression to sexual activity, pregnancy, and sexually transmitted infection among adolescents. However, little information is available about moderators and mediators of these effects. We also know little about digital media, their sex-related content, and their potential influence on youth. Data from a few studies of older youth indicate that sexual displays on social media sites are related to problematic beliefs and behaviors among those who post this content and among viewers. Online pornography appears to be more problematic for youth than off-line sources. Given the vast and increasing amount of time youth spend online and their developmental openness to influence, more research attention to digital sexual media is needed. Those who undertake this work should identify potential negative consequences of use and opportunities to improve adolescent sexual health through digital media. Studies of on- and off-line media in which researchers examine younger media audiences, identify processes explaining sexual media effects on behavior, and moderators of effects are needed. Such studies could be used to inform interventions to reduce negative outcomes and increase positive media effects. Policy makers should stimulate the development of such interventions, including tools to help parents identify and manage negative media influences on their children's sexual well-being and development and dissemination of innovative media literacy programs related to sexual health.


Tissue damage and nutritional factors in experimental respiratory tract (Co-)carcinogenesis.
Cofactors involved in respiratory tract carcinogenesis were studied in Syrian golden hamsters or in rats using benzo(a)pyrene as the carcinogenic agent. These factors included severe tissue damage induced by electro-coagulation, glass fibers administered by intratracheal instillation, acetaldehyde as irritant vapor, food restriction, and nutrients such as vitamin A and saturated and unsaturated fats. In addition, the effects of a combined exposure to four different major gaseous cigarette smoke components--methyl nitrate, isoprene, methyl chloride and acetaldehyde--and to one solid cigarette smoke component--norharman--were examined in short- and long-term inhalation studies. An interesting finding was the carcinogenicity of acetaldehyde, of which the possible mechanism is briefly discussed. Another conspicuous observation was the substantial increase in number and size of lipid droplets in alveolar fibroblasts of hamsters fed a high vitamin A diet.


Antigen-specific cytotoxic T cells mediate human fetal pancreas allograft rejection in SCID-hu mice.
Human allograft rejection was studied in SCID mice transplanted with human fetal liver and thymus tissue (SCID-hu mice). These SCID-hu mice have functional, mature T cells with a polyclonal TCR repertoire. Within 12 to 36 wk after construction, SCID-hu mice were transplanted with an HLA-mismatched human fetal pancreas. In contrast to control SCID mice transplanted with pancreas alone, cellular infiltration, induction of HLA-DR on pancreatic epithelial cells, and tissue destruction of the allogenic pancreata were observed in SCID-hu mice. In addition, human insulin was not detected in the serum of SCID-hu mice in which pancreas rejection occurred. The infiltrating cells were mainly human CD3+ T lymphocytes of thymic origin, expressing the CD45RO isoform. T cell lines and CD4+ T cell clones obtained from the rejected tissues proliferated vigorously when stimulated with EBV-transformed B cell lines of pancreas donor origin. Furthermore, the majority of these CD4+ T cell clones displayed strong allospecific cytotoxicity. In addition, CD8+ T cell clones cytotoxic for EBV-transformed B cell lines of pancreas donors were isolated. Blocking experiments with anti-HLA mAbs and panel studies with HLA-matched cell lines showed that these CD4+ and CD8+ T cell clones were specific for the HLA class II and class I molecules, respectively, expressed by the pancreas donor. These data indicate that human T lymphocytes developing in SCID-hu mice are able to mount in vivo responses against allogenic organs, resulting in tissue infiltration and rejection. In addition, they show that both CD4(+)- and CD8(+)-allospecific CTL can be isolated from rejected allogenic pancreata.


Impact of effective nursing interventions to the fatigue syndrome in children who receive chemotherapy.
This experimental, randomized controlled study was conducted for children with cancer who are 7 to 12 years of age and receiving chemotherapy treatment to detect the impact of appropriate nursing interventions on decreasing the fatigue syndrome. The research sample is composed of a total of 60 children with cancer, with 30 children being included in the experimental group and 30 children included in the control group with their mothers. In the experimental group, after the 7th to 10th day of the chemotherapy treatment, throughout a week, the researcher conducted the effective nursing interventions every day for 45 to 60 minutes. In the control group, routine nursing interventions were carried out. The experimental and control group children's mean scores for the Fatigue Scale-Child and those of mothers for Fatigue Scale-Parent were compared. A statistically significant difference was found between the Fatigue Scale-Child and Fatigue Scale-Parent mean scores of the experimental and the control group children (P < .00). These results suggest that fatigue of children with cancer can be reduced by implementing appropriate nursing interventions.


Rates of prostate surgery and acute urinary retention for benign prostatic hyperplasia in men treated with dutasteride or finasteride.
Previous studies have suggested a greater benefit for various outcomes in men diagnosed with benign prostatic hyperplasia (BPH) who are treated with dutasteride than for men treated with finasteride. This study investigates whether the rates of BPH-related prostate surgery and acute urinary retention (AUR) differ between dutasteride and finasteride users in the Netherlands. From the PHARMO Database Network, men aged ≥50 years with a dispensing of dutasteride or finasteride with or without concomitant alpha-blocker treatment between March 1, 2003 and December 31, 2011 were selected. The incidence of BPH-related prostate surgery and AUR was determined during dutasteride or finasteride treatment and stratified by type of initial BPH-treatment (5-ARI monotherapy or combination with alpha-blocker) and prescriber (general practitioner (GP) or urologist). Comparison of the incidence of BPH-related prostate surgery and AUR between the treatment groups was done by Cox proportional hazard regression. 11,822 dutasteride users and 5,781 finasteride users were identified. Most users started treatment in combination with an alpha-blocker. Overall, dutasteride users had a lower risk of BPH-related prostate surgery was lower among dutasteride users than finasteride users (HR: 0.75; 95 % CI: 0.56-0.99). This lower risk among dutasteride users was also seen when stratifying by monotherapy or combination therapy (HR: 0.73; 95 % CI: 0.54-0.98 for monotherapy and HR: 0.85; 95 % CI: 0.74-0.97 for combination therapy). However, the association was only present among men treated by urologists. For AUR the rates were low and no statistical significant difference was observed between dutasteride and finasteride users. The risk of undergoing BPH-related prostate surgery was lower among men using dutasteride compared to men using finasteride. The association was observed for monotherapy as well as combination therapy, however, only among men who received their prescription from a urologist.


Performance of a portable whole-body mouse exposure system.
A mobile whole-body exposure system was developed for exposing mice to concentrated ambient particulate matter smaller than 2.5 microm in mass median aerodynamic diameter (MMAD). Each 20-L exposure cage was designed to hold 9 mice within individual compartments. This allowed for transport and subsequent exposure. Airflow mixing and the potential for stagnant areas within the compartments were modeled using computational fluid dynamic modeling (CFD). CFD analysis showed no stagnant areas and good mixing throughout the exposure cage. The actual performance of the exposure system was determined for 0.5 to 2.0 microm diameter aerosols by measuring (1) uniformity of aerosol distribution and (2) particle deposition in the tracheobronchial and pulmonary regions of mice exposed in the system. A 0.6-microm MMAD (GSD=2.0) cigarette smoke aerosol was used to experimentally measure the uniformity of aerosol distribution to the nine individual compartments. The average data from three runs showed no statistically significant difference among individual compartments. Particle deposition efficiency in adult male BALB/c mice was measured after exposure (30 min) in the system using monodisperse fluorescent polystyrene latex particles (0.5, 1, and 2 microm aerodynamic diameter). The measured deposition efficiency in this mobile exposure system for the combined tracheobronchial and pulmonary regions of the adult male BALBc mice was 21% for 0.5 microm, 11% for 1.0 microm, and 6.5% for 2.0 microm particles. These deposition efficiencies are similar to those reported for mice exposed in a nose-only exposure system, which indicates that particle losses to animal fur and exposure system surfaces were acceptable.


Diverse balances of tubulin interactions and shape change drive and interrupt microtubule depolymerization.
Microtubules are stiff biopolymers that self-assemble via the addition of GTP-tubulin (αβ-dimer bound to GTP), but hydrolysis of GTP- to GDP-tubulin within the tubules destabilizes them toward catastrophically-fast depolymerization. The molecular mechanisms and features of the individual tubulin proteins that drive such behavior are still not well-understood. Using molecular dynamics simulations of whole microtubules built from a coarse-grained model of tubulin, we demonstrate how conformational shape changes (i.e., deformations) in subunits that frustrate tubulin-tubulin binding within microtubules drive depolymerization of stiff tubules via unpeeling "ram's horns" consistent with experiments. We calculate the sensitivity of these behaviors to the length scales and strengths of binding attractions and varying degrees of binding frustration driven by subunit shape change, and demonstrate that the dynamic instability and mechanical properties of microtubules can be produced based on either balanced or imbalanced strengths of lateral and vertical binding attractions. Finally, we show how catastrophic depolymerization can be interrupted by small regions of the microtubule containing undeformed dimers, corresponding to incomplete lattice hydrolysis. The results demonstrate a mechanism by which microtubule rescue can occur.


Influence of renal sympathetic nerve stimulation on renal function in the primate.
In anesthetized Macaca fascicularis monkeys renal sympathetic nerves were stimulated electrically while changes in renal function were measured. The range of stimulation frequencies was 0.25-4.0 Hz. In the primate the stimulation threshold for changes in renal vascular resistance at a renal perfusion pressure of 100 mmHg was between 1 and 2 Hz. Vascular resistance increased progressively at higher stimulation frequencies. The threshold for changes in sodium reabsorption was somewhat lower (between 0.5 and 1 Hz). At higher stimulation frequencies sodium excretion was dramatically reduced. Renin secretion was not significantly affected at 0.25 and 0.5 Hz stimulation and rose significantly only at stimulation frequencies of 1.0 Hz or above. Thus renal sympathetic nerves in the primate appear to influence renal vascular resistance and electrolyte excretion in a manner that is quantitatively similar to that seen in other species. However, in the primate it was not possible to stimulate renin secretion in the absence of changes in renal blood flow and sodium reabsorption.


Nanowires and nanostructures that grow like polymer molecules.
Unique properties (e.g., rubber elasticity, viscoelasticity, folding, reptation) determine the utility of polymer molecules and derive from their morphology (i.e., one-dimensional connectivity and large aspect ratios) and flexibility. Crystals do not display similar properties because they have smaller aspect ratios, they are rigid, and they are often too large and heavy to be colloidally stable. We argue, with the support of recent experimental studies, that these limitations are not fundamental and that they might be overcome by growth processes that mimic polymerization. Furthermore, we (i) discuss the similarities between crystallization and polymerization, (ii) critically review the existing experimental evidence of polymer-like growth kinetic and behavior in crystals and nanostructures, and (iii) propose heuristic guidelines for the synthesis of "polymer-like" crystals and assemblies. Understanding these anisotropic materials at the boundary between molecules and solids will determine whether we can confer the unique properties of polymer molecules to crystals, expanding them with topology, dynamics, and information and not just tuning them with size.


Electron microscopic study on the effects of different preservation methods for meniscal cartilage.
The study was conducted to evaluate different methods for preservation of meniscal cartilage. During the last decade, the use of allografts containing cartilage in various forms, such as: small osteochondral grafts, osteoarticular grafts, or whole joints, has gained popularity. Meniscal allografts in particular, can be used in cases of post-menisectomy syndrome, or in uni-compartmental osteoarthritis of the knee. The preservation methods used for the cartilaginous components of allografts would appear to influence the success rates, which vary between 30% and 85% according to different reports. Some of the methods of preservation currently in use include: deep-freezing in liquid nitrogen, freezing to -80 degrees C, gamma irradiation, and immersion in ethylene oxide. In an attempt to find the best technique for cartilage preservation (in terms of preserving the ultra-structural properties prior to implantation), bovine meniscal cartilage was examined after preservation by: a) freezing to-80 degrees C; b) exposure to 2.5 Mrad gamma irradiation; or c) a combination of both. Electron microscopic studies were performed, with fresh menisci serving as controls. Our tests demonstrated that in all techniques, severe cellular damage was caused, whereas the collagen network retained its properties. The optimal method for the preservation of cartilage remains to be established.


Patients With Invasive Ductal Carcinoma Have Reduced Levels of Decorin Expression in Their Breast Tissue Compared to Patients With Fibroadenoma While Plasma Decorin Remains Unchanged.
Reduction in the level of tissue decorin is a hallmark of many types of cancer including breast carcinoma. However, reduced decorin expression has also been reported in several types of benign tumors to the extent that it has been proposed as a tissue marker to differentiate malignant from benign tumors. The aim of this study was to investigate the potential role of plasma decorin to distinguish breast cancer from fibroadenoma, the second most common type of benign tumor, after fibrocystic disease. From 35 patients recruited in this study, 24 were affected with invasive ductal carcinoma, either grade II (n = 14) or grade III (n = 10). The other 11 patients had fibroadenoma lesions in their breasts. Tissue decorin mRNA and protein levels were assessed with real-time qPCR and Immunohistochemical analysis. ELISA was employed to measure plasma levels of decorin. The mean plasma decorin in cancer patients was measured to be 5.42 ± 1.83 ng/mL while fibroadenoma patients had an average of 4.22 ± 1.17 ng/mL decorin in their plasma. The difference was not significant. However, the mean expression level of decorin mRNA calculated by the 2-ΔΔCt method was 5.6-fold lower in the biopsied tissue specimens of IDC patients versus fibroadenoma, as expected. Consistent reduction in protein abundance was observed in the studied tissue sections. We have shown that tissue decorin is a reliable marker, unaffected by patient disease stage, to differentiate IDC from fibroadenoma. However, plasma decorin does not seem to have diagnostic value in this regard.


Efficacy of a Goal Setting and Implementation Planning Intervention on Firefighters' Cardiorespiratory Fitness.
Bycura, DK, Dmitrieva, NO, Santos, AC, Waugh, KL, and Ritchey, KM. Efficacy of a Goal Setting and Implementation Planning Intervention on Firefighters' Cardiorespiratory Fitness. J Strength Cond Res 33(11): 3151-3161, 2019-The purpose of this study was to assess the effectiveness of a goal setting and implementation planning (GSIP) intervention on cardiorespiratory fitness among firefighters. Male career firefighters (N = 20) from a large municipal fire department were assigned to an intervention arm (n = 12) or passive control arm (n = 8) of a 14-week study involving cardiorespiratory exercise. The intervention consisted of GSIP coaching at baseline and support throughout the study. Cardiorespiratory fitness was assessed through a standardized physical protocol for firefighters (Candidate Physical Ability Test) at baseline, at 6 weeks, and at 14 weeks time points, using the Cosmed K4b portable metabolic system to measure physiological variables. Using intention-to-treat principles, we employed a 2-level multilevel model to examine the effect of intervention group on intercept and change over time on primary outcomes of interest: oxygen transport and utilization (VO2), heart rate (HR), and respiratory exchange ratio (R). Results demonstrated that the intervention and control groups both exhibited a significant increase in VO2 (B = 0.143, SE = 0.025, p < 0.001) and a decline in R (B = -0.003. SE = 0.001, p < 0.01) over the course of the study. Although both groups exhibited improvements on 2 of the 3 cardiorespiratory outcomes, subjects assigned to the intervention did not exhibit superior outcomes when compared to the control group. Given the improvements we observed in 2 out of 3 outcomes across both conditions, future research may consider testing effectiveness of interventions consisting of functional training exercises that closely translate to firefighting tasks over time periods sufficient to elicit aerobic metabolic adaptations.


Guanidinylated neomycin delivers large, bioactive cargo into cells through a heparan sulfate-dependent pathway.
Facilitating the uptake of molecules into living cells is of substantial interest for basic research and drug delivery applications. Arginine-rich peptides have been shown to facilitate uptake of high molecular mass cargos into cells, but the mechanism of uptake is complex and may involve multiple receptors. In this report, we show that a derivative of the aminoglycoside antibiotic neomycin, in which all of the ammonium groups have been converted into guanidinium groups, can carry large (>300 kDa) bioactive molecules across cell membranes. Delivery occurs at nanomolar transporter concentrations and under these conditions depends entirely on cell surface heparan sulfate proteoglycans. Conjugation of guanidinoneomycin to the plant toxin saporin, a ribosome-inactivating agent, results in proteoglycan-dependent cell toxicity. In contrast, an arginine-rich peptide shows both heparan sulfate-dependent and -independent cellular uptake. The high selectivity of guanidinoneomycin for heparan sulfate suggests the possibility of exploiting differences in proteoglycan compositions to target delivery to different cell types.


Biotinylation of silicon and nickel surfaces and detection of streptavidin as biosensor.
The availability of metal mesh device sensors has been investigated using surface-modified nickel mesh. Biotin was immobilized on the sensor surfaces consisting of silicon and nickel via a thiol-ene click reaction, known as the Michael addition reaction. Biotinylation on the maleimidated surface was confirmed by X-ray photoelectron spectroscopy. The binding of streptavidin to the biotinylated surfaces was evaluated using a quartz crystal microbalance and a metal mesh device sensor, with both techniques providing similar binding constant value. The recognition ability of the biotin immobilized using the thiol-maleimide method for streptavidin was comparable to that of biotin immobilized via several other methods. The adsorption of a biotin conjugate onto the streptavidin-immobilized surface via the biotin-streptavidin-biotin sandwich method was evaluated using a fluorescent microarray, with the results demonstrating that the biological activity of the streptavidin remained.


RALA complexed α-TCP nanoparticle delivery to mesenchymal stem cells induces bone formation in tissue engineered constructs in vitro and in vivo.
A range of bone regeneration strategies, from growth factor delivery and/or mesenchymal stem cell (MSC) transplantation to endochondral tissue engineering, have been developed in recent years. Despite their tremendous promise, the clinical translation and future use of many of these strategies is being hampered by concerns such as off target effects associated with growth factor delivery. Therefore the overall objective of this study was to investigate the influence of alpha-tricalcium phosphate (α-TCP) nanoparticle delivery into MSCs using an amphipathic cell penetrating peptide RALA, on osteogenesis in vitro and both intramembranous and endochondral bone formation in vivo. RALA complexed α-TCP nanoparticle delivery to MSCs resulted in an increased expression of bone morphogenetic protein-2 (BMP-2) and an upregulation in a number of key osteogenic genes. When α-TCP stimulated MSCs were encapsulated into alginate hydrogels, enhanced mineralization of the engineered construct was observed over a 28 day culture period. Furthermore, the in vivo bone forming potential of RALA complexed α-TCP nanoparticle delivery to MSCs was found to be comparable to growth factor delivery. Recognizing the potential and limitations associated with endochondral bone tissue engineering strategies, we then sought to explore how α-TCP nanoparticle delivery to MSCs influences early mineralization of engineered cartilage templates in vitro and their subsequent ossification in vivo. Despite accelerating mineralization of engineered cartilage templates in vitro, RALA complexed α-TCP nanoparticle delivery did not enhance endochondral bone formation in vivo. Therefore the potential of RALA complexed α-TCP nanoparticle delivery appears to be as an alternative to growth factor delivery as a single stage strategy for promoting bone generation.


Direct and indirect effects of impulsivity traits on drinking and driving in young adults.
Impulsivity is strongly associated with alcohol-related risk-taking behavior, and this association has been found to be mediated by alcohol cognitions. The current study expanded this literature by comparing the relative association of distinct impulsivity traits with a specific risky behavior--drinking and driving. We then tested whether drinking-and-driving expectancies uniquely mediated this relation over and above other cognitions about alcohol and drinking and driving. College student drivers (n = 816; 53.6% women) completed a paper-and-pencil survey in small groups. Self-report measures assessed alcohol use, impulsivity traits, alcohol expectancies, drinking-and-driving cognitions (i.e., expectancies, attitudes, beliefs), and drinking and driving. Although all impulsivity traits were correlated with drinking and driving, only urgency uniquely contributed to drinking and driving. Indirect effect tests indicated that drinking-and-driving convenience expectancies partially mediated this association as well as that between (lack of) perseverance and drinking and driving. These results remained significant after controlling for alcohol expectancies and other drinking-and-driving cognitions. These findings highlight the importance of distinguishing among impulsivity traits to improve theoretical models of the processes by which personality leads to specific alcohol-related consequences. In addition, results extend previous research by providing evidence for the unique importance of expectancies regarding the convenience of drinking and driving over and above more global alcohol expectancies and other drinking-and-driving cognitions.


An evaluation of the reliability of time sampling versus continuous observation data collection.
Thirty (10 male, 20 female) children, ages 41 to 71 months, were observed simultaneously in a freeplay setting by two observers using a validated assessment of physical activity in children (i.e. the FATS). One observer utilized a whole interval time sampling method of data recording and the second observer used a hand-held computer and continuously entered observations whenever a child's activity level changed, while the computer simultaneously calculated elapsed time (in tenths of seconds) spent in each behavioral category. Overall activity scores generated by the two approaches correlated 0.90 although the time sampling method produced lower estimates of physical activity. It is concluded that whole interval time sampling is a reliable cost-effective method of assessing physical activity levels in children and may be preferable to the continuous computer-assisted approach that was utilized.


RsaI polymorphism at the cytochrome P4502E1 locus and risk of hepatocellular carcinoma.
CYP2E1, the coding gene for the ethanol inducible cytochrome P4502E1, is polymorphic at the RsaI restriction site in the 5' flanking region. The mutant allele c2 has a higher transcriptional activity than the wild-type gene c1. P4502E1 catalyses the activation of several environmental carcinogens at a rate that is increased, if only moderately, by longterm ethanol intake. To establish the distribution of CYP2E1 RsaI polymorphism in patients with hepatocellular carcinoma and to evaluate its possible role in the multifactorial pathogenesis of this tumour. 101 (84 males) patients with hepatocellular carcinoma and 178 (128 males) healthy controls of the same ethnic (white) and Spanish origin. After extraction of DNA from white blood cells, alleles c1 and c2 of CYP2E1 were identified by restriction fragment length polymorphism (RFLP) with endonuclease RsaI. Homozygous c1c1: 90 patients and 169 controls; heterozygous c1c2: 11 and 9; homozygous c2c2: none (non-significant difference). C2 allele frequencies: 0.055 in patients, 0.025 in controls (non-significant difference) and 0.108 in the 37 patients who had drunk more than 50 g of ethanol/day (p = 0.0035, odds ratio versus controls: 4.67; 95% confidence limits 1.57 to 13.81). The carrier state of one copy of the c2 CYP2E1 gene increases the risk of hepatoma in previously regular ethanol users with chronic liver disease.


Genome-Wide Identification and Characterization of the Biosynthesis of the Polyamine Gene Family in Citrus unshiu.
Polyamines (PAs) contribute to diverse plant processes, environmental interaction, and stress responses. In citrus, the mechanism underlying the biosynthesis of polyamines is poorly understood. The present study aims to identify the biosynthesis of PA gene family members in satsuma mandarin (Citrus unshiu) and investigate their response against various stresses. The identified biosynthesis of PA genes in C. unshiu showed clustering in six groups, i.e., SPMS, SPDS, ACL5, ADC, ODC, and SAMDC. Syntenic analysis revealed that segmental duplication was prevalent among the biosynthesis of PA genes compared to tandem duplication. Thus, it might be the main reason for diversity in the gene family in C. unshiu. Almost all biosynthesis of PA gene family members in C. unshiu showed syntenic blocks in the genome of Arabidopsis, Citrus sinensis, Poncirus trifoliata, and Citrus reticulata. Analysis of Cis-regulatory elements (CREs) indicated the occurrence of hormones, light, defense, and environmental stress responses as well as the development and other plant mechanisms-related elements in the upstream sequence of the biosynthesis of PA genes. Expression profiling revealed that the biosynthesis of PA gene expression modulates in different organs during various developmental stages and stress in C. unshiu. This information will provide a deep understanding of genomic information and its expression in multiple tissues to better understand its potential application in functional genomics.


Giving Your Electronic Health Record a Checkup After COVID-19: A Practical Framework for Reviewing Clinical Decision Support in Light of the Telemedicine Expansion.
The transformation of health care during COVID-19, with the rapid expansion of telemedicine visits, presents new challenges to chronic care and preventive health providers. Clinical decision support (CDS) is critically important to chronic care providers, and CDS malfunction is common during times of change. It is essential to regularly reassess an organization's ambulatory CDS program to maintain care quality. This is especially true after an immense change, like the COVID-19 telemedicine expansion. Our objective is to reassess the ambulatory CDS program at a large academic medical center in light of telemedicine's expansion in response to the COVID-19 pandemic. Our clinical informatics team devised a practical framework for an intrapandemic ambulatory CDS assessment focused on the impact of the telemedicine expansion. This assessment began with a quantitative analysis comparing CDS alert performance in the context of in-person and telemedicine visits. Board-certified physician informaticists then completed a formal workflow review of alerts with inferior performance in telemedicine visits. Informaticists then reported on themes and optimization opportunities through the existing CDS governance structure. Our assessment revealed that 10 of our top 40 alerts by volume were not firing as expected in telemedicine visits. In 3 of the top 5 alerts, providers were significantly less likely to take action in telemedicine when compared to office visits. Cumulatively, alerts in telemedicine encounters had an action taken rate of 5.3% (3257/64,938) compared to 8.3% (19,427/233,636) for office visits. Observations from a clinical informaticist workflow review included the following: (1) Telemedicine visits have different workflows than office visits. Some alerts developed for the office were not appearing at the optimal time in the telemedicine workflow. (2) Missing clinical data is a common reason for the decreased alert firing seen in telemedicine visits. (3) Remote patient monitoring and patient-reported clinical data entered through the portal could replace data collection usually completed in the office by a medical assistant or registered nurse. In a large academic medical center at the pandemic epicenter, an intrapandemic ambulatory CDS assessment revealed clinically significant CDS malfunctions that highlight the importance of reassessing ambulatory CDS performance after the telemedicine expansion.


Prevalence of stenoocclusive lesions in the renal and abdominal arteries in moyamoya disease.
The extracranial involvement of moyamoya disease has been reported in several studies. The main purpose of this study was to determine the prevalence of stenoocclusive lesions in the renal and major abdominal arteries in moyamoya disease. Abdominal angiography was performed in 73 patients with idiopathic moyamoya disease. The findings of abdominal angiography were retrospectively reviewed for the presence and appearance of stenosis in the renal and other major abdominal arteries. Four (5%) of 73 patients presented with unilateral renal artery stenosis. Three patients had moderate stenosis, and one patient had mild stenosis. In the three patients with moderate stenosis, the renal artery stenosis was located in the proximal region of the main branch. Two patients (3%) with moderate stenosis of the unilateral renal artery had renovascular hypertension. No statistically significant differences were observed in age, sex, and the cerebral angiographic stage between patients with and without renal artery stenosis. No stenosis was found in the abdominal aorta or celiac, superior mesenteric, common hepatic, splenic, and proximal common iliac arteries. No occlusions were found in any abdominal arteries. The prevalence of renal artery stenosis in patients with moyamoya disease was 5% (4/73). Involvement of the proximal region of the renal artery was dominant. No stenosis was found in other abdominal arteries.


Circulating tPA contributes to neurovascular coupling by a mechanism involving the endothelial NMDA receptors.
The increase of cerebral blood flow evoked by neuronal activity is essential to ensure enough energy supply to the brain. In the neurovascular unit, endothelial cells are ideally placed to regulate key neurovascular functions of the brain. Nevertheless, some outstanding questions remain about their exact role neurovascular coupling (NVC). Here, we postulated that the tissue-type plasminogen activator (tPA) present in the circulation might contribute to NVC by a mechanism dependent of its interaction with endothelial N-Methyl-D-Aspartate Receptor (NMDAR). To address this question, we used pharmacological and genetic approaches to interfere with vascular tPA-dependent NMDAR signaling, combined with laser speckle flowmetry, intravital microscopy and ultrafast functional ultrasound in vivo imaging. We found that the tPA present in the blood circulation is capable of potentiating the cerebral blood flow increase induced by the activation of the mouse somatosensorial cortex, and that this effect is mediated by a tPA-dependent activation of NMDAR expressed at the luminal part of endothelial cells of arteries. Although blood molecules, such as acetylcholine, bradykinin or ATP are known to regulate vascular tone and induce vessel dilation, our present data provide the first evidence that circulating tPA is capable of influencing neurovascular coupling (NVC).