Graft-versus-host disease prophylaxis with anti-T-cell monoclonal antibody OKT3, prednisone and methotrexate in allogeneic bone-marrow transplantation.
A new regimen for prevention of acute graft-versus-host disease (GvHD)--OKT3 (murine monoclonal anti-pan-T antibody), prednisone and methotrexate (OKT3-pred-MTX)--was compared with the Minnesota standard regimen--antithymocyte globulin, prednisone and methotrexate (ATG-pred-MTX)--for adverse effects, effect on incidence of acute GvHD, and survival at 1 year post-transplant. Twenty patients (aged 25 +/- 9 years) had bone-marrow transplantation (BMT) from their HLA-MLC identical sibling donors for treatment of aplastic anaemia (four), acute leukaemia in remission (13) or chronic myelogenous leukaemia (three). These 20 patients received (OKT3-pred-MTX) on days 8-22 post-transplant. Results of this group are compared to those of 19 concurrent patients (aged 26 +/- 12 years) who received ATG-pred-MTX on days 8-22 post-transplant. On the first day of treatment, 20/20 OKT3 patients and 18/19 ATG patients were febrile. Within 24 h of the first dose of OKT3, 6/20 patients experienced dyspnoea or chest pain and 3/20 patients developed diarrhoea. No further adverse effects were seen after the second dose of OKT3 and no late adverse effects were attributed to this drug. Time to engraftment (means 25 d) was not statistically significantly different in the two prophylactic groups. Acute GvHD was diagnosed in 14 of 20 patients who received OKT3-pred-MTX and in eight of 19 patients who received ATG-pred-MTX (P = 0.06). The incidence of hepatic or gastrointestinal GVHD (greater than or equal to grade 2) was similar in the two groups: 4/20 OKT3-pred-MTX, 6/19 ATG-pred-MTX. Characteristics of post-transplant infections were also similar for the two prophylactic groups. Survival at 1 year post-transplant was 65% for patients who received OKT3 and 44% for patients who received ATG (P = 0.13). The use of OKT3 with prednisone and methotrexate is relatively safe and is associated with a similar incidence of moderate-severe acute GvHD to that experienced in the use of ATG with prednisone and methotrexate.


Advances in the genetic characterization of the malaria vector, Anopheles funestus, and implications for improved surveillance and control.
Anopheles mosquitoes present a major public health challenge in sub-Saharan Africa; notably, as vectors of malaria that kill over half a million people annually. In parts of the east and southern Africa region, one species in the Funestus group, Anopheles funestus, has established itself as an exceptionally dominant vector in some areas, it is responsible for more than 90% of all malaria transmission events. However, compared to other malaria vectors, the species is far less studied, partly due to difficulties in laboratory colonization and the unresolved aspects of its taxonomy and systematics. Control of An. funestus is also increasingly difficult because it has developed widespread resistance to public health insecticides. Fortunately, recent advances in molecular techniques are enabling greater insights into species identity, gene flow patterns, population structure, and the spread of resistance in mosquitoes. These advances and their potential applications are reviewed with a focus on four research themes relevant to the biology and control of An. funestus in Africa, namely: (i) the taxonomic characterization of different vector species within the Funestus group and their role in malaria transmission; (ii) insecticide resistance profile; (iii) population genetic diversity and gene flow, and (iv) applications of genetic technologies for surveillance and control. The research gaps and opportunities identified in this review will provide a basis for improving the surveillance and control of An. funestus and malaria transmission in Africa.


MicroRNAs: New players in cancer prevention targeting Nrf2, oxidative stress and inflammatory pathways.
miRNAs are endogenous small non-coding RNAs of 20-22 nucleotides that repress gene expression at the post-transcriptional level. There is growing interest in the role of miRNAs in cancer chemoprevention, and several naturally occurring chemopreventive agents have been found to be modulators of miRNA expression both in vitro and in vivo. Moreover, these chemopreventive phytochemicals commonly possess anti-oxidative and/or anti-inflammatory properties, and Nrf2 has been extensively studied as a molecular target in cancer prevention. The crosstalk between miRNAs and the traditional cellular signaling pathways of chemoprevention remain to be fully elucidated. This review summarizes the data regarding the potential interactions between miRNAs and anti-oxidative and anti-inflammatory pathways. Cellular redox homeostasis can affect the biogenesis and processing of miRNAs, which in turn regulate the Nrf2 pathway of detoxifying/anti-oxidative genes. We also discuss the miRNA regulatory mechanisms in relation to inflammation-related cancer signaling pathways.


Isolation of keratinophilic fungi from floor dust in Arab elementary and preparatory schools in the West Bank of Jordan.
Floor dust collected from classrooms of thirty three elementary schools (16 for girls, and 17 for boys) (children aged 6-11), and twenty four preparatory schools (13 for girls, and 11 for boys) (children aged 12-14) was studied for the occurrence of keratinophilic fungi with respect to human presence and age of children. Tichophyton mentagrophytes was present in 15.4% of the preparatory schools for girls, in 12.5% of elementary schools for girls, and in 11.8% of elementary schools for boys. It was not found in preparatory schools for boys. Microsporum gypseum was found in preparatory schools for girls only (7.7%). Trichophyton terrestre was also only isolated from elementary schools for boys (5.9%). Chrysosporium species were present in 30.3% of all elementary schools (10 schools/33), and in 20.8% of all preparatory schools (5 schools/24). Geotrichum candidum was the most frequent and predominant keratinophilic species in all schools. Pathogenic and potentially pathogenic keratinophilic fungi comprised a large proportion of all fungal isolates recovered from all schools; they comprised 87.2%-89.5% of all fungal isolates in the elementary schools, and 90.4%-93.5% of all fungal isolates in preparatory schools.


Role of sigmoidoscopy in screening for colorectal cancer: a critical review.
The use of sigmoidoscopy as a screening method for colorectal cancer is controversial. Evidence regarding its efficacy is reviewed critically, with special attention given to potential biases in screening studies. The vast majority of studies are uncontrolled and without follow-up information and thus shed little light on the actual benefits of sigmoidoscopy. Two uncontrolled studies with follow-up and one randomized trial suggest a colorectal cancer mortality reduction because of the use of sigmoidoscopy, but all three studies have major shortcomings. The authors conclude that the currently available data are insufficient to establish a national recommendation for screening with sigmoidoscopy. To establish such a recommendation, a properly conducted randomized trial with colorectal cancer mortality as an outcome is needed.


ANO1-Mediated Inhibition of Cancer Ferroptosis Confers Immunotherapeutic Resistance through Recruiting Cancer-Associated Fibroblasts.
The application of immunotherapy in gastrointestinal (GI) cancers remains challenging because of the limited response rate and emerging therapeutic resistance. Combining clinical cohorts, multi-omics study, and functional/molecular experiments, it is found that ANO1 amplification or high-expression predicts poor outcomes and resistance to immunotherapy for GI cancer patients. Knocking-down or inhibiting ANO1 suppresses the growth/metastasis/invasion of multiple GI cancer cell lines, cell-derived xenograft, and patient-derived xenograft models. ANO1 contributes to an immune-suppressive tumor microenvironment and induces acquired resistance to anti-PD-1 immunotherapy, while ANO1 knockdown or inhibition enhances immunotherapeutic effectiveness and overcomes resistance to immunotherapy. Mechanistically, through inhibiting cancer ferroptosis in a PI3K-Akt signaling-dependent manner, ANO1 enhances tumor progression and facilitates cancer-associated fibroblast recruitment by promoting TGF-β release, thus crippling CD8+ T cell-mediated anti-tumor immunity and generating resistance to immunotherapy. This work highlights ANO1's role in mediating tumor immune microenvironment remodeling and immunotherapeutic resistance, and introduces ANO1 as a promising target for GI cancers' precision treatment.


Predispersal seed predation on five Piper species in tropical rainforest.
Absolute number of seeds lost to predispersal seed predators and proportion of total seeds lost per infructescence were compared among five Costa Rican Piper species of different annual fecundities. Mean seed number and mean seed size in the five species were negatively correlated. The impact of predation on these species was inversely related to the number of seeds they produced. The two early successional species had very high fecundities, a combination of many seeds per infructescence, many infructescences per plant, and, in one species, year-round reproduction. Although seed predators destroyed as many or more seeds of these early successional species than they did of the less fecund, late successional species, this loss accounted for a relatively minor proportion (9 and 12%) of the seeds of the early successional species. In contrast, late successional species produced fewer, larger seeds in a smaller number of infructescences and were not continually in fruit. One of these species, which produced intermediate numbers of intermediately sized seeds, lost 30% of the seeds in each infructescence on average. Seed predators destroyed a larger proportion (65 and 76%) of the seeds per infructescence in the two species with fewest seeds per infructescence. High levels of insect damage in these late successional species caused many of their infructescences to abort prematurely. Taken together these factors resulted in annual fecundities several orders of magnitude smaller in shade-tolerant Piper species than the annual fecundities of shade-intolerant, early successional species. Seedlings of the two early successional species were common in large gaps and other sunny clearings and seedlings of the species with 30% seed loss were occasional, whereas no seedlings were seen of the two species with the highest proportional seed loss, suggesting that seed predation on the latter species may limit seedling recruitment.


The effect of immersion disinfection of elastomeric impressions on the surface detail reproduction of improved gypsum casts.
This investigation examined improved gypsum casts for surface roughness and line-detail reproduction after the immersion disinfection of elastomeric impression materials in an acid glutaraldehyde, an alkaline glutaraldehyde, and a phenol. Impressions were made of a surface roughness standard (Ra = 3.08 microns) that was custom made to include engraved grooves following American Dental Association specification No. 19. Mean surface roughness (Ra) values for all casts of all combinations of disinfectant treatments, impression materials, and improved gypsum stones were obtained with a surface analyzer. Untreated impressions served as controls. Data examined by an analysis of variance indicated that the addition silicone and polyether impression materials provided a surface roughness similar to the precision displacement specimen standard. The acid glutaraldehyde disinfectant demonstrated enhanced line-detail reproduction compared with the standard. Addition silicone and polyether impression materials combined with the acid glutaraldehyde provided the model system closest to the mean surface roughness of the reference standard. These combinations revealed differences in the surface roughness reproduction among the represented improved dental stones.


Interaction of pemetrexed disodium (ALIMTA, multitargeted antifolate) and irradiation in vitro.
Pemetrexed disodium (Alimta, multitargeted antifolate, LY231514; Eli Lilly and Co., Indianapolis, Indiana) ("pemetrexed") is a new folate antimetabolite with significant antitumor activity. Different from classic antifolates, pemetrexed inhibits several key enzymes of thymidylate and purine synthesis, but a radiosensitizing potential may also be presumed. Therefore, the interaction of pemetrexed and ionizing radiation was studied for in vitro clonogenic survival of different human tumor cell lines. Human colon (Widr), breast (MCF-7), cervix (Hela), and lung (LXI) carcinoma cells from log-phase cultures were exposed to pemetrexed (2 h) in combination with different radiation doses given 1 h before pemetrexed washout (all cell lines) or at different points of time before or after pemetrexed addition (Widr). Survival curves were analyzed according to the linear-quadratic (LQ) model, and mean inactivation doses (MID) and radiation enhancement ratios were calculated from the survival curve parameters. Cell-cycle progression of serum-stimulated and pemetrexed- or mock-treated Widr cells was monitored by flow cytometry. Radiosensitization was found for all cell lines at moderately toxic pemetrexed exposures (0.05-0.3 microg/ml [106-636 nM]), but this was cell-type dependent and was most pronounced at roughly isotoxic concentrations, for the least pemetrexed-sensitive Widr cells. Enhancement ratios ranged from about 1.2 (MCF-7 and Hela) to 1.8 (Widr), with a tendency to increase with pemetrexed concentration. Little, if any, change of radiosensitization was observed (Widr) when the time of irradiation was varied from 4 h before to 10 h after the beginning of pemetrexed treatment. Cell-cycle progression of serum-stimulated Widr cells was only marginally affected by pemetrexed. Pemetrexed enhances radiation-induced cell inactivation at moderately toxic exposures and over many hours after drug removal. This effect is not due to disturbed cell-cycle progression, but likely involves an interaction of pemetrexed with long-lived (>4 h) cellular radiation damage and needs to be considered when introducing a combined clinical application.


Personalized medicine using omics approaches in acute respiratory distress syndrome to identify biological phenotypes.
In the last decade, research on acute respiratory distress syndrome (ARDS) has made considerable progress. However, ARDS remains a leading cause of mortality in the intensive care unit. ARDS presents distinct subphenotypes with different clinical and biological features. The pathophysiologic mechanisms of ARDS may contribute to the biological variability and partially explain why some pharmacologic therapies for ARDS have failed to improve patient outcomes. Therefore, identifying ARDS variability and heterogeneity might be a key strategy for finding effective treatments. Research involving studies on biomarkers and genomic, metabolomic, and proteomic technologies is increasing. These new approaches, which are dedicated to the identification and quantitative analysis of components from biological matrixes, may help differentiate between different types of damage and predict clinical outcome and risk. Omics technologies offer a new opportunity for the development of diagnostic tools and personalized therapy in ARDS. This narrative review assesses recent evidence regarding genomics, proteomics, and metabolomics in ARDS research.


EFFECTS OF PANRETINAL PHOTOCOAGULATION ON CHOROIDAL THICKNESS AND CHOROIDAL BLOOD FLOW IN PATIENTS WITH SEVERE NONPROLIFERATIVE DIABETIC RETINOPATHY.
To evaluate the choroidal thickness and choroidal blood flow in the subfoveal region quantitatively after panretinal photocoagulation (PRP) in eyes with severe nonproliferative diabetic retinopathy. This was a prospective comparative study of 24 eyes of 24 patients with type II diabetes and severe nonproliferative diabetic retinopathy with no macula edema. The foveal retinal thickness and choroidal thickness were measured by enhanced depth imaging optical coherence tomography. The subfoveal choroidal blood flow was represented by the mean blur rate obtained by laser speckle flowgraphy. The intraocular pressure, blood pressure, pulse rate, and hemoglobin A1c level (HbA1c) were also measured before and after PRP. The mean foveal retinal thickness did not change significantly during the follow-up period. The mean subfoveal choroidal thickness was reduced significantly from 327.4 μm at the baseline to 286.3 μm at 1 month and 285.0 μm at 3 months after PRP. The mean blur rate ratio decreased significantly to 87.5% at 1 month and 86.0% at 3 months of the baseline values. There was a significant correlation between the subfoveal choroidal thickness and subfoveal choroidal blood flow after PRP. After PRP, the best-corrected visual acuity, intraocular pressure, mean arterial pressure, ocular perfusion pressure, pulse rate, and HbA1c did not change significantly. The success of PRP in treating eyes with severe nonproliferative diabetic retinopathy is probably due to the significant reduction of the subfoveal choroidal thickness and subfoveal choroidal blood flow after PRP.


The ctnF gene is involved in citrinin and pigment synthesis in Monascus aurantiacus.
The application of Monascus is restricted by citrinin. So, it is important to explore the synthetic pathway of citrinin to completely inhibit the production of citrinin. In our previous study, we found that the protein encoded by the ctnF gene has a significant similarity to fructose-2,6-bisphosphatase (F26BPase). It is generally known that the bifunctional enzyme F26BPase regulates the glycolytic flux. So, we speculated that the CtnF protein strengthens carbon flux towards acetyl-CoA and malonyl-CoA which are precursor compounds in citrinin and pigment synthesis. In this study, the ctnF gene-targeting vector pctnF-HPH was constructed and transformed into Monascus aurantiacus. A ctnF-deficient strain was selected by four sets of primers and polymerase chain reaction amplification. Compared with the wild-type strain, citrinin content in the deficient strain was reduced by 34%, and the pigment production was decreased by 72%. These results indicate that the ctnF gene is involved in the common synthesis of citrinin and pigment, which is consistent with previous speculations.


Parenting stress in mothers of children with intractable epilepsy.
The goal of the work described here was to determine the prevalence and characteristics of maternal stress in families of children with intractable epilepsy. Mothers of children aged 2-18 with intractable epilepsy were asked to complete the Parenting Stress Index, Scales of Independent Behavior-Revised, and Child Behavior Checklist, and were queried regarding family type, maternal education, and family income. Neurology clinic charts were reviewed for seizure variables including age at onset, number of failed therapies, and seizure frequency. The Parenting Stress Index measures stress in two broad domains [stress related to characteristics of the child (Child Domain) and stress related to characteristics of the parent (Parent Domain)] as well as a Total Stress score. Fifty-two of 80 (65%) eligible mothers returned completed questionnaires. Sixty-three percent scored in the clinical range for Total Stress, 75% for the Child Domain, but only 29% for the Parent Domain. Mothers scored more adversely on the Isolation, Health, Role Restriction, and Spouse subscales of the Parent Domain, but more favorably on the Attachment subscale. A moderate to high correlation was noted between behavior problems in the child and higher Total Stress scores, but no significant correlations were found between other seizure or demographic variables. Intractable childhood epilepsy is associated with markedly increased maternal parenting stress. Increased stress is due predominantly to child factors. Mothers would strongly benefit from added support to alleviate the constant caregiving demands.


Connective tissue graft plus resin-modified glass ionomer restoration for the treatment of gingival recession associated with non-carious cervical lesions: microbiological and immunological results.
It was previously reported the clinical results of placing subgingival resin-modified glass ionomer restoration for treatment of gingival recession associated with non-carious cervical lesions. The aim of this study was to evaluate the influence of this treatment on the subgingival biofilm and gingival crevicular fluid (GCF) inflammatory markers. Thirty-four patients presenting the combined defect were selected. The defects were treated with either connective tissue graft plus modified glass ionomer restoration (CTG+R) or with connective tissue graft only (CTG). Evaluation included bleeding on probing and probing depth, 5 different bacteria targets in the subgingival plaque assessed at baseline, 45, and 180 days post treatments, and 9 inflammatory mediators were also assessed in the GCF. The levels of each target bacterium were similar during the entire period of evaluation (p > 0.05), both within and between groups. The highest levels among the studied species were observed for the bacterium associated with periodontal health. Additionally, the levels of all cyto/chemokines analyzed were not statistically different between groups (p > 0.05). Within the limits of the present study, it can be concluded that the presence of subgingival restoration may not interfere with the subgingival microflora and with GCF inflammatory markers analyzed. This approach usually leads to the placement of a subgingival restoration. There is a lack of information about the microbiological and immunological effects of this procedure. The results suggest that this combined approach may be considered as a treatment option for the lesion included in this study.


Safety and effectiveness of newer antidiabetic medications during Ramadan fasting and safety of Ramadan fasting after bariatric surgery.
Fasting during Ramadan is mandatory for all adult healthy Muslims. International studies found that most Muslims with diabetes mellitus fast during Ramadan. The main risk factors are hypoglycemia, Hyperglycemia, diabetic ketoacidosis, and dehydration during fasting. Therefore, stratification of the risks for severe acute diabetes complications needs to be considered for each individual and strategies personalized to advert these complications. The advent of new diabetes medications which are effective yet with a better safety profile and monitoring of blood glucose levels during the day are important to reduce the risk of untoward effects of hypoglycemia and hyperglycemia during Ramadan fasting. Here we review the safety and effectiveness of the newer diabetes medications for Ramadan fasting and whether it is safe to perform fasting after bariatric surgery. An extensive literature search using PubMed and Google Scholar was done using different search terms. The eligible studies were 48 randomized controlled trials, prospective observational studies, and reviews from January 2008 to June 2022 which were conducted in individuals living with diabetes. The newer diabetes medications such as GLP-1 agonists, DPP-4 inhibitors, SGLT-2 inhibitors, and new Insulin therapy are thought to be safe and effective during fasting of Ramadan. These medications are associated with a reduction in HbA1c, body weight, systolic blood pressure and risk of hypoglycemia during Ramadan fasting. However, further studies with larger sample size are needed to confirm the efficacy and safety of these newer medications during Ramadan fasting. Individuals with Bariatric surgery should seek advice and approval to fast from the bariatric dietician, physician, and surgeon before the beginning of the month of Ramadan.


In osteoporosis or osteopenia, exercise interventions improve BMD; effects vary by exercise type and BMD site.
Zhang S, Huang X, Zhao X, et al. Effect of exercise on bone mineral density among patients with osteoporosis and osteopenia: a systematic review and network meta-analysis. J Clin Nurs. 2021. [Epub ahead of print]. 34725872.


Modern and traditional approaches combined into an effective gray-box mathematical model of full-blood acid-base.
The acidity of human body fluids, expressed by the pH, is physiologically regulated in a narrow range, which is required for the proper function of cellular metabolism. Acid-base disorders are common especially in intensive care, and the acid-base status is one of the vital clinical signs for the patient management. Because acid-base balance is connected to many bodily processes and regulations, complex mathematical models are needed to get insight into the mixed disorders and to act accordingly. The goal of this study is to develop a full-blood acid-base model, designed to be further integrated into more complex human physiology models. We have developed computationally simple and robust full-blood model, yet thorough enough to cover most of the common pathologies. Thanks to its simplicity and usage of Modelica language, it is suitable to be embedded within more elaborate systems. We achieved the simplification by a combination of behavioral Siggaard-Andersen's traditional approach for erythrocyte modeling and the mechanistic Stewart's physicochemical approach for plasma modeling. The resulting model is capable of providing variations in arterial pCO2, base excess, strong ion difference, hematocrit, plasma protein, phosphates and hemodilution/hemoconcentration, but insensitive to DPG and CO concentrations. This study presents a straightforward unification of Siggaard-Andersen's and Stewart's acid-base models. The resulting full-blood acid-base model is designed to be a core part of a complex dynamic whole-body acid-base and gas transfer model.


Disseminated Mycobacterium abscessus subsp. massiliense infection in a Good's syndrome patient negative for human immunodeficiency virus and anti-interferon-γ autoantibody: a case report.
Good's syndrome (GS) is characterized by immunodeficiency, and can lead to severe infection, which is the most significant complication. Although Mycobacterium rarely causes infection in patients with GS, disseminated nontuberculous mycobacterial (NTM) infection frequently occurs in GS patients that are also positive for the human immunodeficiency virus (HIV) or anti-interferon (IFN)-γ autoantibodies. Here, we report a rare case of GS with NTM without HIV or IFN-γ autoantibodies. A 57-year-old Japanese male with GS and myasthenia gravis (treated with prednisolone and tacrolimus) was diagnosed with disseminated NTM infection caused by Mycobacterium abscessus subsp. massiliense. He presented with fever and back pain. Blood, lumbar tissue, urine, stool, and sputum cultures tested positive for M. abscessus. Bacteremia, spondylitis, intestinal lumber abscess, and lung infection were confirmed by bacteriological examination and diagnostic imaging; urinary and intestinal tract infections were suspected by bacteriological examination but not confirmed by imaging. Despite multidrug combination therapy, including azithromycin, imipenem/cilastatin, levofloxacin, minocycline, linezolid, and sitafloxacin, the patient ultimately died of the infection. The patient tested negative for HIV and anti-IFN-γ autoantibodies. Since myasthenia gravis symptoms interfere with therapy, patients with GS and their physicians should carefully consider the antibacterial treatment options against disseminated NTM.


Changes in thrombomodulin level in plasma of endotoxin-infused rabbits.
Changes in the plasma thrombomodulin (TM) level were examined in endotoxin-infused rabbits. The plasma TM level in normal rabbits was 143.8 +/- 8.4 ng/ml (n = 67) and the molecular weight of the major TM was about 55 kd. Endotoxin (lipopolysaccharide, LPS, E. Coli B8:0127) was intravenously infused. LPS infusion increased the plasma TM level dose-dependently between 0.2 mg/kg and 5 mg/kg. When 5 mg/kg LPS was infused, the plasma TM level started to increase immediately and was 2.3 times higher than the control value within 1 hr. The molecular weight of the major TM was about 75 kd. This rapid increase in TM occurred before the decrease in fibrinogen content and the prolongation of prothrombin time. To examine the effect of circulating leukocytes on the TM increase in endotoxin-infused rabbits, 5 mg/kg LPS was infused into rabbits with leukocytopenia induced by X-ray irradiation. The maximum plasma level of TM was significantly lower than in the untreated rabbits given LPS. These data suggest that the increase in plasma TM is caused by LPS-stimulated leukocyte's prior to hemostaseological changes. It is well known that endothelial cells can be injured by stimulated leukocytes, so this increase in plasma TM probably reflects the deterioration of endothelial cells. This deterioration decreases the ability of endothelial cells to inhibit thrombosis, which would, in turn, contribute to the development of disseminated intravascular coagulation in endotoxin-infused rabbits.


Biological effects of a murine retrovirus carrying an activated N-ras gene of human origin.
We have introduced a genomic DNA clone of a mutated human N-ras gene from a T-cell leukemia cell line into a retroviral vector equipped with a neo resistance gene and with SV40 and pBR322 origins of replication. The helper free N-ras virus, which was recovered after transfection of the construction in the psi 2 packaging cell line, contained a correctly spliced N-ras gene. Proviral DNA was amplified in cos cells and subsequently cloned in bacteria. Nucleic acid sequence analysis of the activated N-ras gene revealed a point mutation at codon 12 resulting in a glycine to aspartic acid substitution. The N-ras virus was able to transform mouse fibroblastic cell lines, but failed to fully transform mouse primary embryo fibroblasts. MoMuLV or amphotropic 4070A pseudotypes of the virus were injected intraperitoneally into newborn mice. The MoMuLV pseudotype produced only helper-virus-induced leukemias. The amphotropic pseudotype caused fibrosarcomas after a long latent period. The results of these and other in vivo experiments are discussed in relation to known pathogenic effects of other retroviruses carrying H-ras or K-ras genes.


A comparative pharmacokinetic study of ethanol in the blood, vitreous humour and aqueous humour of rabbits.
The pharmacokinetics of ethanol in the blood, vitreous humour and aqueous humour of the rabbit after intravenous administration of single dose have been compared. Ethanol levels were determined by gas-liquid chromatography. Certain parallelism between ethanol levels in these body fluids were observed after 80 min.


Slow degrading Mg-based materials induce tumor cell dormancy on an osteosarcoma-fibroblast coculture model.
Osteosarcoma is one of the most common cancers in young adults and is commonly treated using surgery and chemotherapy. During the past years, these therapy approaches improved but failed to ameliorate the outcomes. Therefore, novel, targeted therapeutic approaches should be established to enhance treatment success while preserving patient's quality of life. Recent studies suggest the application of degradable magnesium (Mg) alloys as orthopedic implants bearing a potential antitumor activity. Here, we examined the influence of Mg-based materials on an osteosarcoma-fibroblast coculture. Both, Mg and Mg-6Ag did not lead to tumor cell apoptosis at low degradation rates. Instead, the Mg-based materials induced cellular dormancy in the cancer cells indicated by a lower number of Ki-67 positive cancer cells and a higher p38 expression. This dormancy-like state could be reversed by reseeding on non-degrading glass slides but could not be provoked by inhibition of the protein kinase R-like endoplasmic reticulum kinase. By investigating the influence of the disjunct surface-near effects of the Mg degradation on cell proliferation, an increased pH was found to be a main initiator of Mg degradation-dependent tumor cell proliferation inhibition.


Coracoclavicular and coracoacromial ligament calcification and ossification.
Thirty-six patients with calcification or ossification at or around the coracoclavicular and coracoacromial regions were analyzed with regard to type, location, and configuration of the deposits and related clinical history. Calcification or ossification in the coracoclavicular region resulted largely from trauma (36%) or renal failure (28%). Trauma patients may develop punctate calcification or ossification but do not develop the tumoral type of calcification. About 5% of the renal failure patients had coracoclavicular ligament calcifications, one-half of which were of the tumoral type. Renal failure patients may have punctate or tumoral calcifications but do not develop ossification.


Modulation of plasma metalloproteinase-9 concentrations and peripheral blood monocyte mRNA levels in patients with septic shock: effect of fiber-immobilized polymyxin B treatment.
The authors measured plasma metalloproteinase (MMP)-9 and corresponding monocyte mRNA in 20 patients with septic shock. Plasma MMP-9 concentrations and monocyte MMP-9 mRNA levels were significantly higher in the 10 nonsurviving patients with septic shock than in 10 surviving patients and 25 normal controls. Hemoperfusion using polymyxin B immobilized on fibers (PMX-F), a reportedly effective treatment for septic shock, was studied for effects on MMP-9 in the patients. Increases in plasma MMP-9 concentrations and corresponding monocyte mRNA levels were attenuated significantly by PMX-F treatment in both nonsurviving and surviving patients. These data suggest that plasma MMP-9 concentrations and monocyte MMP-9 mRNA levels may be useful prognostic markers in septic shock, and that PMX-F treatment affects MMP-9.


Aqua regia digestion cannot completely extract Hg from biochar: A synchrotron-based study.
Mercury (Hg) is commonly extracted from solid phase samples using aqua regia for total Hg (tHg) analysis. However, uncertainties exist regarding the complete extraction of Hg by aqua regia, especially from carbonaceous materials. To investigate whether aqua regia can completely extract Hg from biochars, batch-style experiments were carried out to evaluate extraction efficiency of aqua regia with respect to Hg-loaded biochar and to characterize the residual Hg speciation and spatial distribution. Different types of biochars (raw, FeCl3-modified, and FeSO4-modified, prepared at different temperatures) were reacted with Hg-spiked solution before the digestion experiments. Adsorption analyses indicate the biochars were successfully loaded with Hg and that the Hg content was higher in biochars pyrolyzed at higher temperature (900 versus 300 or 600 °C). The results of digestion experiments indicate Hg could not be completely extracted from the biochars tested, with a greater percentage of residual Hg in biochars pyrolyzed at 600 (60 ± 15%) and 900 (75 ± 22%) than 300 °C (7 ± 2%). Furthermore, the fraction of residual Hg in FeSO4-modified biochars after aqua regia digestion was significantly lower than in FeCl3-modified and unmodified biochars. Confocal micro-X-ray fluorescence imaging (CMXRFI) showed residual Hg in biochars is concentrated on surfaces prior to digestion, but more homogeneously distributed after digestion, which indicates Hg on biochar surface is more easily digested. Hg extended X-ray absorption fine structure (EXAFS) spectra modelling showed residual Hg in biochars mainly exists as Hg(II)-Cl. These results indicate extra caution should be paid for tHg determinations using aqua regia digestion method in soil (especially in forest), sediment, and peat samples containing black carbon, activated carbon, or biochar.


Patent iliolumbar artery increase no risk of type II endoleaks after endovascular abdominal aortic aneurysm: a case-control study.
The aims of the present study were to explore the risk factors for type 2 endoleaks (T2ELs) after endovascular aneurysm repair (EVAR) and the association between T2ELs and the iliolumbar artery. A single-center, retrospective case-control study in West China Hospital was conducted among patients with infrarenal abdominal aortic aneurysm (AAA) who underwent EVAR between June 2010 and June 2019. The associations of patient characteristics, anatomical factors, internal iliac artery embolization, and ILA with the primary outcome were analyzed. The secondary objective was to analyze survival and reintervention between the T2EL group and the non-T2EL group. Kaplan-Meier survival, propensity matching analysis and multivariate logistic regression analysis were used. A total of 603 patients were included. The median follow-up was 51 months (range 5.0-106.0). There was a significant difference in the diameter of the lumbar artery (LA), middle sacral artery (MSA) and inferior mesentery artery (IMA), proportion of thrombus and LA numbers. The univariate analysis showed that T2ELs were more likely to develop more thrombus in aneurysm cavity (OR = 0.294, p = 0.012), larger MSA (OR = 1.284, p = 0.04), LA (OR = 1.520, p = 0.015), IMA (OR = 1.056, p < 0.001) and more LAs (OR = 1.390, p = 0.019). The multivariate analysis showed that the number of LAs (HR: 1.349, 95% CI: 1.140-1.595, p < .001) and the diameter of the IMA (HR: 1.328, 95% CI: 1.078-1.636, p = 0.008) were significantly associated with T2ELs. There were no new findings from the propensity score matching. The reintervention-free survival rates were significantly different between the two groups (p = 0.048). Overall survival and AAA-related death rates were not different between the two group. This was consistent with the PSM analysis. The iliolumbar artery and the different internal iliac artery interventions may not increase the incidence of T2ELs. But the numbers of LAs and IMA diameter were independent risk factors for T2Els. T2ELs was associated with the reintervention but did not affect long-term survival or increase aneurysm-related mortality after EVAR.


Identification of an additional hypoxia responsive element in the glyceraldehyde-3-phosphate dehydrogenase gene promoter.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional enzyme overexpressed in many tumors and induced by hypoxia in normal and malignant cells. The degree to which hypoxia transcriptionally activates GAPDH is cell type specific. The GAPDH promoter region contains a hypoxia responsive element (HRE) consisting of a hypoxia inducible factor-1 (HIF-1) consensus binding site plus adjacent sequence [Graven et al. (1999) Biochim. Biophys. Acta 1447, 208-218]. Using transient transfection experiments with the GAPDH promoter region linked to a luciferase reporter gene, we found that GAPDH was transcriptionally activated by hypoxia in each of three human prostate cancer cell lines tested, with the greatest level of induction in the most differentiated cell line. Using sequence analysis of the GAPDH promoter region, we identified a novel HRE distinct from the previously characterized one that consists of two consensus HIF-1 sites arranged as inverted repeats separated by 5 bp. Hypoxia transcriptionally activated a promoter construct in which the previously characterized HRE was mutated and the novel HRE remained intact. Heterologous promoter constructs containing only one or two copies of the novel HRE plus a minimal promoter consisting of a TATA box drove hypoxia inducible expression of the luciferase reporter gene in transient transfection assays. Mutation of HIF-1 sites within the novel HRE resulted in complete loss of function.


Spinal cord transection: a quantitative analysis of elements of the connective tissue matrix formed within the site of lesion following administration of piromen, cytoxan or trypsin.
Long-Evans hooded rats were cordotomized at the T-5 level and given either (1) cyclophosphamide (cytoxan), an immunosuppressive, (2) piromen, a bacterial polysaccharide-nucleic acid complex, (3) topical and systemic trypsin, or (4) no further specific treatment. Because of past and present controversy surrounding the proposed ability of these agents to promote spinal cord regeneration, a systematic study, employing light and electron microscopy, and quantitative methods in a single animal model, was done in order to re-evaluate the effects of each treatment upon the connective tissue matrix which forms in the defect left by transection. After an initial inflammatory reaction during the first week after surgery, the lesion zone is characterized either by areas of dense collagenous connective tissue with occasional fibroblasts and macrophages, or a loose areolar tissue with numerous sheets and cords of mesodermal cellular elements but minimal collagen. By 45 days postoperatively (dpo), axons supported by Schwann cells invade and become entangled in the loose connective tissue matrix. With longer postoperative survival, cysts appear craniad and caudad to the lesion and erode much of the scar together with viable neural tissue. Giving cytoxan or piromen did not result in any qualitative alteration of the scar matrix as evidenced by electron microscopy. Quantitative analysis revealed a slight reduction in the fibrous connective tissue component of the scar at 45--90 dpo, but this was transient when longer postoperative periods were studied. Trypsin caused a significant reduction in the amount of fibrous connective tissue with a concomitant increase in loose connective tissue and the appearance of a few distinctive, compact bundles of unmyelinated axons lacking Schwann cells. Consistent behavioral changes were not observed in any group which could distinguish them from the controls. Our results appear to contradict the findings of Matinian and Andreasian (1976) who reported return of normal sensori-motor function in 80% of their animals treated with topical and systemic trypsin. It is concluded that a major impediment to whatever longterm regenerative potential exists within the spinal cord is the lack of axonal guiding elements within the scar, but more importantly, the severe erosion of the remaining spinal cord due to cyst enlargement.


[Immunologic and morphologic characteristics of experimental infection with Mycoplasma pneumoniae].
Pathomorphology of experimental M. pneumoniae infection in 3-week-old guinea pigs was characterized by an early cytopathic action of the causative agent on the bronchial and the alveolar epithelium, and also by progressive infiltrative-proliferative changes in the bronchovascular and interstitial tissue of the lungs against the background of morphological reconstruction in the immunogenesis organs. Immunological and histomorphological study of reinfected guinea pigs pointed to the presence of delayed hypersensitivity features in the response cellular-tissue reactions.


Neuropsychiatric manifestations of Sydenham's chorea: a systematic review.
Sydenham's chorea is a post-streptococcal, autoimmune, neuropsychiatric movement disorder. Sydenham's chorea is a major criterion for diagnosis of acute rheumatic fever with the implication of potential long-term sequelae including cardiac complications. It is well established that there is psychiatric comorbidity in Sydenham's chorea, but there are variations in the literature regarding the nature and prevalence of psychiatric diagnoses associated with Sydenham's chorea. The aim of this review was to systematically evaluate the evidence for psychiatric symptoms presenting with Sydenham's chorea. Knowledge of comorbid psychiatric symptomatology will support early diagnosis and treatment, leading to improved long-term outcomes for children with Sydenham's chorea. The study used a systematic search strategy, using MEDLINE, MEDLINE in Process, EMBASE, and The Cochrane Library. Abstracts were screened to identify relevant papers which were then assessed further. Eligible papers were summarized. A total of 1429 abstracts of relevant studies were found, and 49 papers reporting neuropsychiatric symptoms in Sydenham's chorea were summarized. Obsessive-compulsive disorder was the most commonly studied, and hence reported, neuropsychiatric symptom in children with Sydenham's chorea. The studies analysed used a variety of tools to identify affected children and used different methods for analysing results. Attention-deficit-hyperactivity disorder, affective disorders, tic disorders, executive function disturbances, and psychotic features were also reported as comorbidities. There is good evidence of neuropsychiatric comorbidities in Sydenham's chorea. In countries with a high prevalence of rheumatic fever, the early recognition of salient cognitive and psychiatric symptoms may aid in the management of Sydenham's chorea.


A supervoxel based random forest synthesis framework for bidirectional MR/CT synthesis.
Synthesizing magnetic resonance (MR) and computed tomography (CT) images (from each other) has important implications for clinical neuroimaging. The MR to CT direction is critical for MRI-based radiotherapy planning and dose computation, whereas the CT to MR direction can provide an economic alternative to real MRI for image processing tasks. Additionally, synthesis in both directions can enhance MR/CT multi-modal image registration. Existing approaches have focused on synthesizing CT from MR. In this paper, we propose a multi-atlas based hybrid method to synthesize T1-weighted MR images from CT and CT images from T1-weighted MR images using a common framework. The task is carried out by: (a) computing a label field based on supervoxels for the subject image using joint label fusion; (b) correcting this result using a random forest classifier (RF-C); (c) spatial smoothing using a Markov random field; (d) synthesizing intensities using a set of RF regressors, one trained for each label. The algorithm is evaluated using a set of six registered CT and MR image pairs of the whole head.


Engraftment syndrome: a common cause for rash and fever following autologous hematopoietic stem cell transplantation for multiple sclerosis.
Autologous hematopoietic stem cell transplantation (HSCT) is currently being evaluated as a therapy for patients with progressive multiple sclerosis (MS) at risk of debilitating neurological impairment. While preliminary results from a few studies have been reported, little is known about toxicities or outcome of HSCT for MS. We report a relatively frequent triad of non-infectious fever, rash and fatigue or lassitude that may also be associated with pruritus, pulmonary symptoms, and eosinophilia and frequently occurs around engraftment. This syndrome occurred in 26% of our series of patients (5/19) undergoing HSCT for multiple sclerosis. The engraftment syndrome is usually self-limited but may require intervention with systemic corticosteroids.


A critical review of analytical methods for determination of curcuminoids in turmeric.
Turmeric (Curcuma longa) is one of the most important ingredients in Indian and Chinese cuisine. Curcuminoids and volatile oils present in turmeric are known for their functional and nutraceutical properties. Health benefits attributed to curcuminoids have resulted in their wide utilization in food and pharmaceutical formulations. Therefore, characterization and estimation of the curcuminoids in fresh/dry turmeric, food and nutraceutical products are essential for their quality control during processing and storage. To meet the demand for analytical methods of curcuminoids, several methods have been developed for their quantification in turmeric powder and food formulations. In the present review, various analytical methods (spectrophotometric, chromatographic, capillary electrophoresis and biosensor techniques) which are used for monitoring curcuminoids have been thoroughly summarized and discussed. The spectrophotometric method is not useful when individual components of curcuminoids are required. Mobile phase optimization, the broadness of spots, plate-to-plate variations are significant limitations for TLC and HPTLC methods. Many analysts believe that HPLC method is the best choice for curcuminoids determination because of its rapid analysis. Spectrofluorimetry and Electrochemical methods are the more advanced methods with high sensitivity as well as rapid analysis. However, the selection of analytical method for curcuminoids analysis depends on the type of sample matrix, purpose of the analysis and limit of detection and limit of quantitation of the method.


Purification of Dengue and Zika Virus Non-structural Protein 5 for Crystallization and Screening of Antivirals.
Dengue Virus (DENV) and ZIKA Virus (ZIKV) are two important human pathogens that belong to the Flavivirus genus of positive strand RNA viruses. Symptoms of DENV infections range from asymptomatic or mild fever to life-threatening forms, while ZIKV can lead to teratogenic effects such as microcephaly in newborns and neurological disease like the Guillain-Barré syndrome.Non-Structural Protein 5 (NS5) is the largest and most conserved enzyme across flaviviruses and hence constitutes a prime target for developing pan-flavivirus antiviral inhibitors. NS5 results from the gene fusion between a methyltransferase at the N-terminus of the protein and an RNA-dependent RNA polymerase (RdRp) at the C-terminal end. The NS5 protein plays key roles in replication and modification of viral RNA and its inhibition by potent antiviral drugs could prevent severe symptoms associated with infections.We have optimized purification and crystallization protocols to obtain active recombinant proteins suitable for structure-based drug discovery for both the full-length NS5 protein and the polymerase domain of NS5 from DENV and ZIKV .


Experimental investigation of H3PO4 activated papaya peels for methylene blue dye removal from aqueous solution: Evaluation on optimization, kinetics, isotherm, thermodynamics, and reusability studies.
This investigation is centered on the effectiveness of methylene blue (MB), a cationic dye, adsorbed from an aqueous media by H3PO4 activated papaya skin/peels (PSPAC), with initial pH (2-10), contact time (30-180 min), MB dye concentration (varying from 10 to 50 mg/L), and MB dose (0.1-0.5 gm). The findings show that the best optimal conditions for MB dye removal occur at a 6 pH, 0.3 gm dose of PSPAC adsorbent for 10 mg/L MB dye concentration, with 90 min of contact time. To optimize and validate the extraction efficiency of MB dye, a response surface methodology (RSM) study was conducted using a central composite design (CCD) with a regression model showing R2 = 0.9940. FT-IR spectroscopy shows, CO, and O-H stretching functional groups while FE-SEM is assessed to supervise morphological features of the PSPAC adsorbent. The peak adsorption capacity with 46.95 mg/g for the Langmuir isotherm model conveniently satisfies the adsorption process with R2 = 0.9984 while with R2 = 0.999, a kinetic model, pseudo-second-order, confirms MB dye adsorption by PSPAC adsorbent. Moreover, thermodynamic parameters including ΔGᵒ, ΔH°, and ΔS° were computed and found to be spontaneous and exothermic. Furthermore, regeneration studies employed with NaOH (0.1 M) and HCl (0.1 M) solution media show an acceptable MB removal efficiency consecutive up to three cycles. The study highlights that H3PO4 papaya skin/peel (PSPAC) is an effectual, sustainable, reasonably available biosorbent to remove industrial cationic dyes disposal.


The impact of depression and sense of coherence on emotional and social loneliness among nursing home residents without cognitive impairment - a questionnaire survey.
To analyse the relationships between depressive symptoms, sense of coherence and emotional and social loneliness among nursing home residents without cognitive impairment. Depression symptoms and loneliness are major health problems for older people. Sense of coherence, which is based on a salutogenic theoretical framework, is a strong determinant of positive health and successful coping and is associated with well-being and depression among older people. Few studies have explored the relationships between depression symptoms, sense of coherence and emotional and social loneliness among nursing home residents. A cross-sectional, descriptive, correlational design. Sample - 227 residents 65-102 years old from 30 nursing home residing ≥ six months. All had a Clinical Dementia Rating ≤ 0·5 and could converse. Residents were interviewed using the Social Provisions Scale, Geriatric Depression Scale and Sense of Coherence Scale (SOC-13). Possible relationships between these were analysed, controlled for sex, age, marital status, education, length of stay and comorbidity. Before adjustment, Geriatric Depression Scale was associated with attachment and social integration. After adjustment, Geriatric Depression Scale was still associated with attachment and social integration. Further adjusting for Sense of Coherence Scale reduced the association between Geriatric Depression Scale and attachment and even more so for the association between Geriatric Depression Scale and social integration. Sense of coherence and Geriatric Depression Scale did not interact, and SOC-13 was associated with attachment and social integration. Depression symptoms contribute to emotional and social loneliness. Independent of sense of coherence, depression symptoms are associated with emotional loneliness, sense of coherence influence emotional and social loneliness. Clinical nurses should observe residents closely for signs of depression and loneliness and support their sense of coherence to reduce emotional and social loneliness.


Expression of recombinant human bifunctional peptidylglycine α-amidating monooxygenase in CHO cells and its use for insulin analogue modification.
The availability of catalytically active peptidylglycine α-amidating monooxygenase (PAM) should provide the means to examine its potential use for the chemienzymatic synthesis of bioactive peptides for the purpose of pharmacological studies. Hypoglycemic activity is one of the most important features of insulin derivatives. Insulin glargine amide was found to show a time/effect profile which is distinctly more flat and thus more advantageous than insulin glargine itself. The aim of the study was to obtain recombinant PAM and use it for insulin analogue amidation. We stably expressed a recombinant PAM in CHO dhfr-cells in culture. Recombinant PAM was partially purified by fractional ammonium sulphate precipitation and ion-exchange chromatography. The enzyme was used to modify glycine-extended A22(G)-B31(K)-B32(R) human insulin analogue (GKR). Alpha-amidated insulin was analyzed by HPLC and mass spectrometry. Hypoglycemic activity of amidated and non-amidated insulin was compared. The pharmacodynamic effect was based on glucose concentration measurement in Wistar rats with hyperglycemia induced by streptozotocin. The overall glycemic profile up to 36 h was evaluated after subcutaneous single dosing at a range of 2.5-7.5 U/kg b.w. The experiment on rats confirmed with a statistical significance (P < 0.05) hypoglycemic activity of GKR-NH2 in comparison to a control group receiving 0.9% NaCl. Characteristics for GKR-NH2 profile was a rather fast beginning of action (0.5-2.0 h) and quite prolonged return to initial values. GKR-NH2 is a candidate for a hypoglycemic drug product in diabetes care. In addition, this work also provides a valuable alternative method for preparing any other recombinant bioactive peptides with C-terminal amidation.


DSM-III personality disorders and family history of mental illness.
This report investigates the way in which the presence of a personality disorder (PD) in probands affects the frequency of psychiatric disorders in relatives. Four proband groups--panic, panic plus a PD, major depression, and depression plus a PD--were given the Family History-Research Diagnostic Criteria and the Family History for DSM-III Anxiety and Personality Disorders. Information was gathered on 730 first-degree relatives. Panic patients with a PD had higher incidences of panic and panic plus a PD in relatives. There was increased depression in relatives of probands with either depression or a PD. Alcoholism appeared highest in relatives of probands who had combined depression and a PD.


Potentiometric investigation of the stability of palladium(II) complex of pralidoxime chloride in aqueous solution.
The formation of a complex between palladium(II) chloride and pralidoxime chloride (PAM-2Cl) has been studied by means of potentiometric pH measurements. The real stability constant of the complex in aqeous medium of ionic strength 0.3 M (KCl) at 25.0 degrees C was log Ks = 7.29. This value was close to that (log Ks = 7.02) obtained previously by spectrophotometric methods after appropriate correction with respect to the corresponding value of the acidic constant of PAM-2Cl (pKca = 8.05), which was also determined under the same experimental conditions.


Differential expression of chaperonin containing T-complex polypeptide (CCT) subunits during fetal and adult skin wound healing.
Integumentary wound healing in early fetal life is regenerative and proceeds without scar formation. Expressomic analysis of this phenomenon by differential display has previously determined that the eta subunit of the cytosolic chaperonin containing T-complex polypeptide (CCT) is downregulated in the healing fetal wound milieu. We now report that no other CCT subunit shares this distinct pattern of gene regulation as determined by limiting dilution reverse transcriptase polymerase chain reaction (RT-PCR); all seven of the remaining CCT subunits demonstrate no change in messenger RNA (mRNA) expression in healing fetal wounds compared to unwounded control tissue. The alpha subunit, however, did evidence reduced message levels in healing adult wound tissue. We herein report on the cloning and sequence of the complementary DNA (cDNA) for rabbit CCT-alpha and confirm its wound specific decrease in adult tissues through quantitative real-time RT-PCR assay. We also confirm that quantitative evaluation of CCT-alpha and CCT-zeta mRNA expression shows no change in healing fetal wounds.


Blood pressure variability: a novel and important risk factor.
Blood pressure is a continuous, not a static, variable. Individuals exhibiting similar clinic or home blood pressure can differ considerably with respect to their average day and nighttime values, beat-by-beat blood pressure variation during wakefulness and sleep, responses to mental and physical stimuli, and intersession and seasonal variation. There now is evidence that several such representations of blood pressure variability, if augmented, increase cardiovascular risk independent of the average of conventionally acquired blood pressure readings. As well, recent retrospective analyses of published trial data have concluded that antihypertensive drug classes differ in their effects on intersession blood pressure variability and associated risk of stroke. If the goal of the hypertension community is to optimize personalized cardiovascular risk assessment and to attenuate fully such risk, future efforts should be directed at determining which representation of blood pressure variability estimates individual cardiovascular risk best, establishing "normal" and "high- risk" variability distributions, testing the hypothesis that attenuating such variability specifically through drug or device therapy reduces cardiovascular risk more than blood pressure reduction per se, and integrating such data into clinical practice.


Filtering effects in reporting work injuries.
Research efforts in the field of occupational injuries have been hampered by difficulties in ascertaining true prevalence rates. Lack of accuracy in reporting and documentation of work injuries leads to loss of information and lack of comparability of work injury statistics. The paper explores some of the factors related to loss of information about work injuries and proposes a descriptive model of the information processing system. The Filter Model describes the way in which information about a given injury proceeds through a series of levels, from true prevalence of injuries in the workplace, to prevalence as indicated by national aggregate statistics. Between each level is a "filter" or barrier that is selectively permeable, ensuring that information about some, but not all, injuries will pass through to the next level. Consequently, the type of work injury data collected at each level will differ. The Filter Model can aid in choice of appropriate level of data for research purposes, and provides a framework for investigation into the nature and extent of filtering processes within work injury reporting systems.


Effects of quorum sensing on the biofilm formation and viable but non-culturable state.
Quorum sensing exists widely in all kinds of microorganisms and is a communication channel for microorganisms. Many bacterial processes, including virulence factor expression, biofilm formation, and viable but non-culturable (VBNC) cell resuscitation, are mediated by quorum sensing, and biofilm formation complicates the treatment of various infections. Foodborne pathogens can enter VBNC state in extreme environments, and pathogens in VBNC state can evade traditional detection and resuscitate under appropriate conditions, causing potential harm to human health. The disruption of quorum sensing may decisively help control biofilm formation and VBNC cell resuscitation. This review describes the quorum sensing systems of various bacteria and major fungi, and summarizes the role of bacterial quorum sensing system in biofilm formation and VBNC resuscitation. In addition, the relationship between quorum sensing inhibitors (QSI) with biofilms and VBNC is also discussed.


Acid deposition in the Athabasca Oil Sands Region: a policy perspective.
Industrial emissions of sulphur (S) and nitrogen (N) to the atmosphere associated with the oil sands industry in north-eastern Alberta are of interest as they represent the largest localized source in Canada (with potential for future growth) and the region features acid-sensitive upland terrain. Existing emission management policy for the Regional Municipality of Wood Buffalo, where the industry is located, is based on a time-to-effect approach that relies on dynamic model simulations of temporal changes in chemistry and features highly protective chemical criteria. In practice, the policy is difficult to implement and it is unlikely that a scientifically defensible estimate of acidification risk can be put forward due to the limitations primarily associated with issues of scale, chemical endpoint designation (selection of chemical limit for ecosystem protection from acidification) and data availability. A more implementable approach would use a steady-state critical load (CL) assessment approach to identify at-risk areas. The CL assessment would consider areas of elevated acid deposition associated with oil sands emissions rather than targeted political jurisdictions. Dynamic models should only be (strategically) used where acidification risk is identified via CL analysis, in order to characterize the potential for acidification-induced changes that can be detrimental to sensitive biota within the lifespan of the industry.


Ionic liquid based on α-amino acid anion and N7,N9-dimethylguaninium cation ([dMG][AA]): theoretical study on the structure and electronic properties.
The interactions between five amino acid based anions ([AA](-) (AA = Gly, Phe, His, Try, and Tyr)) and N7,N9-dimethylguaninium cation ([dMG](+)) have been investigated by the hybrid density functional theory method B3LYP together with the basis set 6-311++G(d,p). The calculated interaction energy was found to decrease in magnitude with increasing side-chain length in the amino acid anion. The interaction between the [dMG](+) cation and [AA](-) anion in the most stable configurations of ion pairs is a hydrogen bonding interaction. These hydrogen bonds (H bonds) were analyzed by the quantum theory of atoms in molecules (QTAIM) and natural bond orbital (NBO) analysis. Finally, several correlations between electron densities in bond critical points of hydrogen bonds and interaction energy as well as vibrational frequencies in the most stable configurations of ion pairs have been checked.


Temporal and spatial assessment of normal cerebrospinal fluid dynamics with MR imaging.
The purpose of this study was to measure normal cerebrospinal fluid (CSF) pulsations within the intracranial and upper cervical subarachnoid spaces and the ventricular system. Phase contrast cine MR sequences were performed in sagittal and axial planes on 13 volunteers with flow encoding in the craniocaudal direction. CSF pulsations displayed considerable variations in healthy subjects, depending both on measurements localization and subjects, with CSF peak velocities ranging from 0 to 7 cm/s. In the subarachnoid spaces, the highest velocities occurred in the anterior location and increased from the cerebellar pontine angle cisterns towards the lower cervical spaces. In the ventricular system, the highest velocities occurred through the aqueduct of Sylvius. CSF flow within the third ventricle seemed to reflect a circular motion. There was a caudal net CSF flow in the aqueduct whereas in the upper cervical spaces net CSF flow was caudal anteriorly and cranial laterally. Velocity profiles of CSF pulsations demonstrated arterial morphology. After the R wave, caudal systolic motion was first observed in the posterior subarachnoid spaces, soon after in the anterior subarachnoid spaces and later in the ventricular system. Considering the morphology of CSF pathways, three successively initiated phenomena may explain the temporal course of CSF motion: the systolic expansion of the main arteries at the base of the brain, the systolic expansion of the cerebrospinal axis and, finally, the systolic expansion of the choroid plexuses.


Genetic and environmental contributions to the expression of handedness in chimpanzees (Pan troglodytes).
Most humans are right-handed and, like many behavioral traits, there is good evidence that genetic factors play a role in handedness. Many researchers have argued that non-human animal limb or hand preferences are not under genetic control but instead are determined by random, non-genetic factors. We used quantitative genetic analyses to estimate the genetic and environmental contributions to three measures of chimpanzee handedness. Results revealed significant population-level handedness for two of the three measures-the tube task and manual gestures. Furthermore, significant additive genetic effects for the direction and strength of handedness were found for all three measures, with some modulation due to early social rearing experiences. These findings challenge historical and contemporary views of the mechanisms underlying handedness in non-human animals.


Genome-wide association studies in aging-related processes such as diabetes mellitus, atherosclerosis and cancer.
Recent technological developments allow to genotype several hundreds of thousands of genetic variants in a single person in one step. This enables genome-wide association studies (GWAS) by genotyping a large number of patients with diseases of interest and controls at reasonable costs. Compared to a hypothesis-driven candidate gene approach the hypothesis-free GWAS can identify new susceptibility genes without making any a priori biological assumptions. They permit to identify genes involved in pathways which until now were unknown to be involved in a certain phenotype. GWAS are therefore a new and very powerful tool to identify genetic contributors to aging-related phenotypes. This paper provides a short overview about design and methods of GWAS and reviews recent advances in the identification of susceptibility genes for type 2 diabetes mellitus, atherosclerosis and cancer using GWAS.


Transcriptional plasticity of different ABC transporter genes from Tribolium castaneum contributes to diflubenzuron resistance.
The development of insecticide resistance challenges the sustainability of pest control and several studies have shown that ABC transporters contribute to this process. ABC transporters are known to transport a large range of chemically diverse molecules across cellular membranes, and therefore the identification of ABC transporters involved in insecticide resistance is difficult. Here, we describe a comprehensive strategy for the identification of whole sets of ABC transporters involved in insecticide resistance using the pest beetle, Tribolium castaneum (Tc) as a model. We analyzed the expression of ABCA to ABCC genes in different tissues and developmental stages using larvae that were sensitive or resistant to diflubenzuron (DFB). The mRNA levels of several ABC genes expressed in excretory or metabolic tissues such as midgut, Malpighian tubules or fat body were markedly upregulated in response to DFB. Next, we monitored mortality in the presence of the ABC inhibitor verapamil, and found that it causes sensitization to DFB. We furthermore established a competitive assay for the elimination of DFB, based on Texas Red (TR) fluorescence. We monitored TR elimination in larvae that were treated with DFB or different ABC inhibitors, and combinations of them. TR elimination was decreased significantly in the presence of DFB, verapamil and the ABCC inhibitor MK-571. The effect was synergized when DFB and verapamil were both present suggesting that the transport of TR and DFB involves overlapping sets of ABC transporters. Finally, we silenced the expression of DFB-responding ABC genes by RNA interference and then followed the survival rates after DFB exposure. Mortality increased particularly when specific ABCA and ABCC genes were silenced. Taken together, we were able to show that different ABC transporters expressed in metabolic and excretory tissues contribute to the elimination of DFB. Up- or down-regulation of gene expression occurs within a few days already at very low DFB concentrations. These results suggests that transcriptional plasticity of several ABC genes allows adaptation of the efflux capacity in different tissues to eliminate insecticides and/or their metabolites.


Screening Yeast Display Libraries against Magnetized Yeast Cell Targets Enables Efficient Isolation of Membrane Protein Binders.
When isolating binders from yeast displayed combinatorial libraries, a soluble, recombinantly expressed form of the target protein is typically utilized. As an alternative, we describe the use of target proteins displayed as surface fusions on magnetized yeast cells. In our strategy, the target protein is coexpressed on the yeast surface with an iron oxide binding protein; incubation of these yeast cells with iron oxide nanoparticles results in their magnetization. Subsequently, binder cells that interact with the magnetized target cells can be isolated using a magnet. Using a known binder-target pair with modest binding affinity (KD ≈ 400 nM), we showed that a binder present at low frequency (1 in 105) could be enriched more than 100-fold, in a single round of screening, suggesting feasibility of screening combinatorial libraries. Subsequently, we screened yeast display libraries of Sso7d and nanobody variants against yeast displayed targets to isolate binders specific to the cytosolic domain of the mitochondrial membrane protein TOM22 (KD ≈ 272-1934 nM) and the extracellular domain of the c-Kit receptor (KD ≈ 93 to KD > 2000 nM). Additional studies showed that the TOM22 binders identified using this approach could be used for the enrichment of mitochondria from cell lysates, thereby confirming binding to the native mitochondrial protein. The ease of expressing a membrane protein or a domain thereof as a yeast cell surface fusion-in contrast to recombinant soluble expression-makes the use of yeast-displayed targets particularly attractive. Therefore, we expect the use of magnetized yeast cell targets will enable efficient isolation of binders to membrane proteins.


Thiolated 2-methacryloyloxyethyl phosphorylcholine for an antifouling biosensor platform.
We developed a new building block for a protein- and cell-repellant self-assembled monolayer (SAM) from 2-methacryloyloxyethyl phosphorylcholine (MPC) via a simple Michael-type addition to one mercapto group in alkanedithiol. The thiolated MPC can enable functionalization of a noble metal electrode to minimize noise signal in biosensing.


Neurofibromatosis-1: a maximum likelihood estimation of mutation rate.
Methods of classical segregation analysis were applied to a sample of 129 sibships with one or more individuals affected by neurofibromatosis-1 (NF-1). The sample consists only of subjects with NF-1; all the probands had been referred for genetic counselling because of café-au-lait spots, and a diagnostic protocol was invariably applied. No deviation from the segregation ratio expected for a fully penetrant Mendelian dominant gene was observed. A maximum likelihood estimate of the proportion of sporadic cases was obtained, and the mutation rate was estimated to be 6.5 x 10(-5) gametes per generation (95% CI 5.0-8.1).


Reconstructing atom probe data: a review.
Atom probe tomography stands out from other materials characterisation techniques mostly due to its capacity to map individual atoms in three-dimensions with high spatial resolution. The methods used to transform raw detector data into a three-dimensional reconstruction have, comparatively to other aspects of the technique, evolved relatively little since their inception more than 15 years ago. However, due to the importance of the fidelity of the data, this topic is currently attracting a lot of interest within the atom probe community. In this review we cover: (1) the main aspects of the image projection, (2) the methods used to build tomographic reconstructions, (3) the intrinsic limitations of these methods, and (4) future potential directions to improve the integrity of atom probe tomograms.


Lipid-lowering therapy and cholesterol levels following acute myocardial infarction: a German study of 5361 patients.
Current studies on secondary prevention of cardiovascular events (CARE, LIPID, 4S) illustrate the necessity of an effective lipid-lowering therapy. An important part of secondary prevention is the prompt measurement of lipids following an infarct, to be able to start therapy as quickly as possible. This study should show the general situation in the prescription of lipid-lowering drugs in patients with acute myocardial infarction (MI) and if there are gender differences in therapy and in the lipid parameters. The second aim was to determine the therapeutical conclusions for secondary prevention made in-hospital in patients with heart attacks. Post-infarct lipid parameters (total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), triglycerides (TG) were examined in 5361 patients within 24 hours following an acute MI. Lipid parameters from 576 patients were measured again after 1 week. 80.1% of men under examination and 81.9% of the women had suffered from their first MI. Only 9% of all patients were already under lipid-lowering therapy before the MI. After the MI TC and LDL-C levels decreased significantly in a time-dependent manner. Around 50% of patients received lipid-lowering drug, or the recommendation of one, during their hospital stay. The range of TC values of the patients examined was comparable to those in the CARE and the 4S secondary prevention studies. It can therefore be assumed that the results of these studies are also applicable to Germany. Nevertheless, according to existing data, therapy with lipid-lowering drugs is currently unsatisfactory, even in secondary prevention.


Blockade of CTLA-4 enhances allergic sensitization and eosinophilic airway inflammation in genetically predisposed mice.
CTLA-4 (CD152) expression is restricted to subsets of activated T lymphocytes and shares homology with CD28. CTLA-4 and CD28 molecules both bind to B7 molecules on antigen-presenting cells. Whereas CD28-B7 interaction enhances T cell activation, cytokine production and survival, CTLA-4 signaling down-regulates T cell responses. Here, we studied the involvement of CTLA-4 triggering in the pathogenesis of allergen-induced airway inflammation in mice. Anti-CTLA-4 mAb were injected during i.p. sensitization with ovalbumin (OVA). This treatment favored OVA-specific IgE production and augmented blood eosinophilia in BALB/c mice. In BALB/c mice, enhanced Th2 sensitization after anti-CTLA-4 mAb injections resulted in more severe airway inflammation, and increased airway hyperresponsiveness to metacholine, bronchial eosinophilia and IL-4 and IL-5 levels in broncho-alveolar lavage (BAL) fluid following repeated allergen inhalations. Importantly, aggravation of airway inflammation and enhancement of Th2 responses were accompanied by a significant reduction of pulmonary TGF-beta levels at protein level in BAL fluid as well as on mRNA level in inflamed lung tissue. In contrast to BALB/c mice, blockade of CTLA-4 did not alter IgE production nor the phenotype of airway inflammation or TGF-beta production in C57BL/6 mice. Our data suggest that CTLA-4 triggering represents an important regulatory mechanism for Th2 sensitization in genetically predisposed mice by modulating TGF-beta production.


Ostreopsis cf. ovata bloom in the northern Adriatic Sea during summer 2009: ecology, molecular characterization and toxin profile.
Intense blooms of the benthic dinoflagellate Ostreopsis cf. ovata have occurred in the northern Adriatic Sea since 2006. These blooms are associated with noxious effects on human health and with the mortality of benthic organisms because of the production of palytoxin-like compounds. The O. cf. ovata bloom and its relationships with nutrient concentrations at two stations on the Conero Riviera (northern Adriatic Sea) were investigated in the summer of 2009. O. cf. ovata developed from August to November, with the highest abundances in September (1.3×10(6) cells g(-1) fw corresponding to 63.8×10(3) cells cm(-2)). The presence of the single O. cf. ovata genotype was confirmed by a PCR assay. Bloom developed when the seawater temperature was decreasing. Nutrient concentrations did not seem to affect bloom dynamics. Toxin analysis performed by high resolution liquid chromatography-mass spectrometry revealed a high total toxin content (up to 75 pg cell(-1)), including putative palytoxin and all the ovatoxins known so far.


The lncOb rs10487505 polymorphism impairs insulin sensitivity and glucose tolerance in children and adolescents with obesity.
Leptin plays a key role in the regulation of body weight and other endocrine systems. Recently, impairment of leptin gene transcription due to genetic variations in a long noncoding RNA (lncOb) has been described. This retrospective study aims to characterize the clinical and metabolic phenotype of children and adolescents with obesity who were homozygous for the lncOb rs10487505 leptin lowering allele. Enrolled children underwent an anthropometrical evaluation, biochemical assessment, and genotyping for lncOb rs10487505. Plasma leptin levels were assessed in 150 participants. A total of 434 patients were included and divided into two groups according to rs10487505 recessive inheritance (CC vs. GG/GC). Children who were homozygous for the C allele showed higher fasting insulin (p = 0.01), homeostasis model assessment of insulin resistance (p = 0.01), lower whole-body insulin sensitivity index (p = 0.02), and lower disposition index (p = 0.03). Moreover, CC patients presented with a higher prevalence of prediabetes (9.3% vs. 3.4%, p = 0.04) and a 2.9-fold (95% CI: 1.1-7.9, p = 0.04) higher risk of prediabetes compared with G-carriers independently from confounders. Leptin plasma levels were significantly lower in the CC group (p = 0.002). Hormone levels correlated with BMI z score (r = 0.19, p = 0.04), fasting insulin (r = -0.34, p < 0.0001), homeostasis model assessment of insulin resistance (r = -0.33, p < 0.0001), and disposition index (r = 0.20, p = 0.04). The lncOb rs10487505 polymorphism affects leptin circulating levels, worsens insulin resistance, and heightens the risk of prediabetes in children and adolescents with obesity.


Right thoracotomy, femorofemoral bypass, and deep hypothermia for re-replacement of the mitral valve.
Ten patients underwent mitral valve re-replacement for the third to sixth time through a right thoracotomy using one-lung anesthesia, femorofemoral bypass, profound systemic hypothermia, and low-flow perfusion without aortic cross-clamping or cardioplegia. The indications for this approach were previous mediastinitis, severe right ventricular hypertension with multiple previous sternotomies, intact coronary artery bypass graft, or previous aortic valve replacement. There was 1 operative death, which was due to end-stage pulmonary hypertension and intractable right heart failure. Blood loss was minimal, and there was no major postoperative morbidity in the 9 surviving patients except for supraventricular arrhythmias.


Anomalies in Human Sex Determination: Usefulness of a Combined Cytogenetic Approach to Characterize an Additional Case with Xp Functional Disomy Associated with 46,XY Gonadal Dysgenesis
Disorders of sexual development (DSD) are a heterogeneous group of genital defects affecting chromosomal, gonadal and anatomical sex. 46,XY DSD is a subset of DSD which covers a wide range of phenotypes in which 46,XY gonadal dysgenesis (GD) is the most severe form. In this study, we report on the clinical and molecular cytogenetic findings of a study on a Tunisian girl with the syndromic form of 46,XY DSD. This case was a phenotypic female patient having several congenital anomalies including growth retardation. Karyotype, fluorescence in situ hybridization and array Comparative Genome Hybridization (array CGH) were performed. The proband exhibited a de-novo 46,X,der(Y) karyotype. Array CGH revealed a pathogenic 27.5Mb gain of an Xp21.2 chromosome segment leading to Xp functional disomy. No deletion was observed in the Y-chromosome. The duplicated region encompassed the NR0B1 (DAX1) and MAGEB genes, located within the dosage sensitive sex (DSS) reversal locus, known as promote genes responsible for human sex reversal and testis repression. The extra-dosage and interactions of these genes with different specific genes could result in the impairment of the male sex pathway. Over-dosage of KAL1 and IL1RAPL1 genes fall within the somatic features observed in the patient. To the best of our knowledge, we report on the fourth case of Xp21.2-pter duplication within Xp;Yp translocation associated with XY GD. Our findings suggest that when duplicated, the NR0B1 and MAGEB genes could be a major cause of XY GD. Therefore, we emphasize the usefulness of a combined cytogenetic approach in order to provide an accurate genetic diagnosis for those patients having syndromic XY DSD in a clinical setting.


New nonsteroidal androgen receptor modulators based on 4-(trifluoromethyl)-2(1H)-pyrrolidino[3,2-g] quinolinone.
A series of 2(1H)-pyrrolidino[3,2-g]quinolinones was prepared and tested for the ability to modulate the transcriptional activity of the human androgen receptor (hAR). The parent compound, 4-(trifluoromethyl)-2(1H)-pyrrolidino[3,2-g]quinolinone, displayed moderate interaction with hAR, but more substituted analogues, particularly 6,7-disubstituted compounds, were potent hAR agonists in vitro.


Is a 22 cm Ureteric Stent Appropriate for Korean Patients Smaller than 175 cm in Height?
Determining the ideal length of a ureteric stent is important to avoid complications associated with stent placement. Clinically, most urologists usually choose the length of a ureteric stent according to the patient's height. On the basis of a Chinese population study, a 22 cm ureteric stent has been recommended for patients smaller than 175 cm. We evaluated the appropriateness of this recommendation in Korean patients. A total of 70 patients who were smaller than 175 cm and who underwent ureteroscopic lithotripsy and ureteric stent insertion were studied. The appropriateness of the stent length was determined on the basis of plain film findings. Patient discomfort was measured by use of a visual analogue scale (VAS) before the removal of the ureteric stent. In 29 patients with a 22 cm ureteric stent, 21 patients (72.4%) had an appropriate ureteric stent length and the mean VAS was 4.1. In 36 patients with a 24 cm ureteric stent, 20 patients (55.6%) had an appropriate ureteric stent length and the mean VAS was 4.0. Among 5 patients with a 26 cm ureteric stent, 1 patient (20%) had an appropriate ureteric stent length and the mean VAS was 5.4. In Korean patients smaller than 175 cm in height, a 22 cm ureteric stent was an appropriate length.


Selection of macronutrients by goldfish operating self-feeders.
In the wild, both food availability and an animal's requirements may fluctuate. Given the choice to feed on different diets, animals may exhibit preferences in their selection. Although macronutrient selection has been investigated in mammals and birds, few attempts have been made to investigate dietary selection in fish. In the present study we investigate the feeding of goldfish on three "pure" macronutrient diets. Twelve single goldfish and one group of four goldfish, initially weighing 163.1 +/- 7.4 and 70.2 +/- 8.0 g, respectively, were maintained in 54-liter tanks at 20 degrees C under a 12 h:12 h light-dark (LD) cycle. Three pelleted diets were made containing 50% of each macronutrient: protein (casein + gelatin), fat (fish oil + soy oil), or carbohydrate (dextrin). All the diets included mixtures of vitamins and minerals, a binder (sodium alginate), and cellulose as a filler. During trials, three self-feeding devices delivering the protein, fat, or carbohydrate diet were installed in each aquarium. Goldfish soon learned to select from among the three diets. Overall daily food and digestible energy demands differed significantly between macronutrients. The demand for protein was lower than that for carbohydrate (16.6 and 33.7 g/kg BW/day, respectively), and the digestible energy demand for protein was lower than for fat (108.6 and 308.5 kJ/kg BW/day, respectively). Goldfish made their dietary selections on the basis of energy content, so that food demand increased to compensate for changes in the digestible energy density of the diets. Correlation analysis revealed a significant positive relationship between carbohydrate and fat demand (n = 8), whereas there were significant negative (n = 3) and positive (n = 1) relationships between protein and fat. Different daily patterns for the selection of each macronutrient were detected in some fish (i.e., carbohydrate during daytime, protein at the beginning of the night, and fat during the transition phase). In short, our results demonstrated the ability of goldfish to compose their own diet from three basic macronutrients, highlighting the complex pattern of behavior in their selection. Therefore, these new findings should be considered when studying feeding rhythms and the design of diets for fish.


Penetration of levofloxacin into skin tissue after oral administration of multiple 750 mg once-daily doses.
To probe the pharmacokinetic basis for the use of levofloxacin for complicated skin and skin-structure infections (SSSIs) at a once-daily dosage of 750 mg by investigating its penetration into skin tissue. Ten healthy volunteers were administered three oral, once-daily 750 mg doses of levofloxacin, and levofloxacin concentrations were subsequently measured over time (0.5-24 h) in skin-punch biopsy tissue and plasma. Skin tissue concentrations consistently exceeded those in plasma at every time point, with tissue/plasma ratios of 1.37 +/- 0.81 for peak concentration and 1.97 +/- 0.35 for area under the concentration versus time curve. Three of the ten subjects reported treatment-emergent adverse events (AEs) that were considered unrelated to treatment. An 11th subject who had enrolled in the study withdrew after AEs of mild severity that were possibly related to the study drug. The results support the clinical usage of levofloxacin 750 mg once-daily for complicated SSSIs.


Erratum: Author Correction. Management of ovarian cancer patients in affected areas during COVID-19 pandemic: Japan and Korea.
This corrects the article on p. e65 in vol. 31, PMID: 32342678.


Pulmonary embolism despite negative ELISA D-dimer: a case report.
Pulmonary embolus (PE) can be fatal, but is often treatable if recognized early. Unfortunately, the clinical presentation of PE is often variable and misleading. The D-dimer assay has recently come into favor as a method to exclude PE; however, this test has an acceptable safety margin only in low-risk populations. What is unclear is the exact composition of this low risk population. This is the report of a 26-year-old woman with over 2 weeks of chest pain and intermittent dyspnea. The patient was initially seen in the Emergency Department (ED) and hospitalized. She returned to the ED 2 weeks later with similar symptoms. Although enzyme-linked immunosorbent assay (ELISA) D-dimer assays were normal on the initial and subsequent ED visits, pulmonary embolism (PE) was diagnosed by computed tomography scan on the second visit. This report highlights the risk of misdiagnosing PE if relying solely on ELISA D-dimer for exclusion. The approach to PE should include a measure of clinical probability. This report documents the presentation of PE despite having two unremarkable ELISA D-dimer measurements, and highlights the importance of clinical suspicion.


Differential effect of BMP4 on NIH/3T3 and C2C12 cells: implications for endochondral bone formation.
After intramuscular implantation, BMP4-expressing NIH/3T3 fibroblasts and BMP4-expressing C2C12 myoblasts can promote ectopic cartilage and bone formation. Fibroblasts tend to undergo chondrogenesis, whereas myoblasts primarily undergo osteogenesis. These results suggest that endochondral bone formation may involve different cell types, a finding that could have major implications for the tissue engineering of bone and cartilage. The delivery of BMP4 through cell-based gene therapy can trigger ectopic endochondral bone formation in skeletal muscle. We hypothesized that, when stimulated with or transduced to express BMP4, different types of cells residing within skeletal muscle might participate in different stages of endochondral bone formation. We compared the responses of a fibroblast cell line (NIH/3T3), a myoblast cell line (C2C12), primary fibroblasts, and primary myoblasts to BMP4 stimulation in vitro. We then transduced the four cell populations to express BMP4 and compared their ability to promote ectopic endochondral bone formation in skeletal muscle. Under the influence of BMP4 in vitro and in vivo, NIH/3T3 cells differentiated toward both chondrogenic and osteogenic lineages, whereas most C2C12 cells underwent primarily osteogenic differentiation. NIH/3T3 cells genetically modified to express BMP4 induced delayed but more robust cartilage formation than did genetically modified C2C12 cells, which promoted rapid ossification. These differences in terms of the timing and amount of cartilage and bone formation persisted even after we introduced a retrovirus encoding dominant negative Runx2 (DNRunx2) into the C2C12 cells, which interferes with the function of Runx2. Superior osteogenic potential was also displayed by the primary myoblasts in vitro and in vivo compared with the primary fibroblasts. The different proliferation abilities and differentiation potentials exhibited by these cells when influenced by BMP4 may at least partially explain the differing roles that BMP4-expressing myogenic cells and BMP4-expressing fibroblastic cells play in endochondral bone formation. Our findings suggest that the process of endochondral bone formation in skeletal muscle after delivery of BMP4 involves different cell types, including fibroblastic cells, which are more involved in the chondrogenic phases, and myoblastic cells, which are primarily involved in osteogenesis. These findings could have important implications for the development of tissue engineering applications focused on bone and cartilage repair.


Clinicopathologic Factors and Their Association with Outcomes of Salivary Duct Carcinoma: A Multicenter Experience.
This series reports long-term clinical outcomes of patients with salivary duct carcinoma (SDC), which is associated with a poor prognosis. Eighty-nine patients with SDC were treated with curative intent from February 5, 1971, through September 15, 2018. Kaplan-Meier and competing risk analyses were used to estimate locoregional control, distant metastasis-free survival (DMFS), progression-free survival, and overall survival (OS). Cox regression analyses of disease and treatment characteristics were performed to discover predictors of locoregional control, DMFS, and OS. Median follow-up was 44.1 months (range, 0.23-356.67). The median age at diagnosis was 66 years (interquartile range, 57-75). Curative surgery followed by adjuvant radiation therapy was performed in 73 patients (82%). Chemotherapy was delivered in 26 patients (29.2%). The 5-year local recurrence and distant metastasis rates were 27% and 44%, respectively, with death as a competing risk. Distant metastasis was associated with lymph node-positive disease (hazard ratio [HR], 3.16; 95% confidence interval [CI], 1.38-7.23; P = .006), stage IV disease (HR, 4.78; 95% CI, 1.14-20.11; P = .033), perineural invasion (HR, 4.56; 95% CI, 1.74-11.97; P = .002), and positive margins (HR, 9.06; 95% CI, 3.88-21.14; P < .001). Median OS was 4.84 years (95% CI, 3.54-7.02). The 5-year OS was 42%. Reduced OS was associated with lymphovascular space invasion (HR, 3.49; 95% CI, 1.2-10.1; P = .022), perineural invasion (HR, 2.05; 95% CI, 1.06-3.97; P = .033), positive margins (HR, 2.7; 95% CI, 1.3-5.6; P = .011), N2 disease (HR, 1.88; 95% CI, 1.03-3.43; P = .04), and N3 disease (HR, 11.76; 95% CI, 3.19-43.3; P < .001). In this single-institution, multicenter retrospective study, the 5-year survival was 42% in patients with SDC. Lymphovascular space invasion, lymph node involvement, and higher staging at diagnosis were associated with lower DMFS and OS.


[Radiological diagnosis of thymoma: consideration on 52 cases (author's transl)].
Review of 52 cases of thymoma (41 typical, 11 atypical) studied radiologically and controlled surgically and anatomo-pathologically. Differential diagnosis is discussed; the importance of pneumomediastinum and angiography is stressed and a number of cases are reported in detail.


A program for simulation of nerve equations with branching geometries.
A computer program has been developed for simulation of electrical activity in neurons with complex branching morphology, multiple channel types, and inhomogeneous channel distribution. The program is based around an interpreter and screen editor for flexible specification of nerve properties and analysis of simulation results. Efficient simulation of the nerve specification is accomplished with procedure calls to fast, compiled routines for integration of the nerve equations.


Comparative Transcriptome Analysis Reveals Regulatory Mechanism of Long Non-Coding RNAs during Abdominal Preadipocyte Adipogenic Differentiation in Chickens.
Long non-coding RNAs (lncRNAs) are implicated in mammalian adipogenesis and obesity. However, their genome-wide distribution, expression profiles, and regulatory mechanisms during chicken adipogenesis remain rarely understood. In the present study, lncRNAs associated with adipogenesis were identified from chicken abdominal adipocytes at multiple differentiation stages using Ribo-Zero RNA-seq. A total of 15,179 lncRNAs were identified and characterized by stage-specific expression patterns. Of these, 840 differentially expressed lncRNAs were detected, and their cis- and trans-target genes were significantly enriched in multiple lipid-related pathways. Through weighted gene co-expression network analysis (WGCNA) and time-series expression profile clustering analysis, 14 key lncRNAs were identified as candidate regulatory lncRNAs in chicken adipogenic differentiation. The cis- and trans-regulatory interactions of key lncRNAs were constructed based on their differentially expressed cis- and trans-target genes, respectively. We also constructed a competing endogenous RNA (ceRNA) network based on the key lncRNAs, differentially expressed miRNAs, and differentially expressed mRNAs. MSTRG.25116.1 was identified as a potential regulator of chicken abdominal preadipocyte adipogenic differentiation by acting as a transcriptional trans-regulator of fatty acid amide hydrolase (FAAH) gene expression and/or a ceRNA that post-transcriptionally mediates FAAH gene expression by sponging gga-miR-1635.


PAI1 inhibits the pathogenesis of primary focal hyperhidrosis by targeting CHRNA1.
Primary focal hyperhidrosis (PFH) may be attributed to the up-regulation of the cholinergic receptor nicotinic alpha 1 subunit (CHRNA1) in eccrine glands. Plasminogen activator inhibitor-1 (PAI1, encoded by SERPINE1) is reported to inhibit the expression of CHRNA1, while the role of PAI1 in hyperhidrosis is unknown. Serpine1 KO mice, Serpine1-Tg mice, and wild type BALB/c mice were intraperitoneally injected with pilocarpine hydrochloride to induce PFH. Cisatracurium (CIS, antagonist of CHRNA1) or PAI-039 (small-molecule inhibitor of PAI1) was pre-administrated before the induction of hyperhidrosis. On the other hand, Chrna1-expressing AAV was constructed and administered to Serpine1-Tg mice with hydrochloride stimulation. Hydrochloride-related biomarkers, such as acetylcholine (ACH) in the serum, calcium voltage-gated channel subunit alpha1 C (CACNA1C), and aquaporin 5 (AQP5) in sweat glands of mice were assayed with ELISA, RT-PCR, and Western blot. The administration of PAI-039 or Pai1 knock-out increased Chrna1 expression, sweat secretion, and hydrochloride-related biomarkers (ACH, CACNA1C, and AQP5) expression. On the other hand, CIS administration diminished the strengthened hyperhidrosis phenotype induced by Pai1 knock-out with decreased sweat gland secretion. PAI1 inhibits CHRNA1-mediated hydrochloride-induced hyperhidrosis, with decreased sweat gland secretion and diminished ACH, AQP5, and CACNA1C expression. These results indicate the potential to utilize PAI1 to alleviate PFH.