The primary season of nitrate leaching in the Loess Plateau region is the monsoon, which is caused by heavy rainfall during the growth season of corn (Zea mays L.). Nitrate leaching to groundwater is an increasing concern in agriculture, and is one of the major nitrogen losing ways in dryland farming system. Localized compaction and ridge fertilization is a method for nitrogen fertilizer application, by which, less fertilizer leaching would occur. The NO3(-)-N transport in soil profile, corn yields and nitrogen use efficiency under localized compaction and ridge fertilization were investigated through two years field study. The factors that affect NO3(-)-N transport under localized compaction and ridge fertilization were studied, combined with simulated experiment. The results showed that NO3(-)-N was leached to below 90 cm in plat fertilization in the year of about 370 mm rainfall, a mean precipitation during the season, while the NO3(-)-N leakage of the fertilizer zone was reduced by localized compaction and ridge fertilization, as a result that the NO3(-)-N concentration below 60 cm was less than 10 mg.kg-1, and NO3(-)-N accumulated in 20-40 cm with a concentration 80-90 mg.kg-1. There was no significant difference in yield between application methods with 240.0 kg N.hm-2. However, the absorbed amount of nitrogen was improved significantly by localized compaction and ridge fertilization, and the nitrogen use efficiency was increased by 9%. The bulk density of the barriers had an evident effect on NO3(-)-N transport under localized compaction and ridge fertilization, but the effect of ridge slope was insignificant.

Gastroduodenal artery aneurysm is a rare vascular lesion, asymptomatic in the majority of cases. However, ruptured aneurysm is associated with poor prognosis and mortality can achieve a 40% rate. We here report the case of an 83-year-old patient with abrupt onset of non-specific abdominal pain associated with hematemesis. Endoscopy showed beating formation compressing the duodenal bulb with active bleeding. Abdominal computed tomography (CT) scan was performed which objectified gastro duodenal artery aneurysms involving the artery ostium and making endovascular treatment impossible to perform. Thus, the patient underwent open surgery based on flattening associated with aneurysm exclusion and then complemented by bulb plasty. Post-operative CT scan confirmed total exclusion of the aneurysm with preservation of hepatic circulation.

L'anévrisme de l'artère gastroduodénale est une lésion vasculaire rare, asymptomatique dans la majorité des cas. Cependant, en cas de rupture de l’anévrisme, le pronostic est mauvais et la mortalité peut atteindre 40%. Nous rapportons le cas d’un patient âgé de 83 ans, qui a présenté brutalement des douleurs abdominales non spécifiques, associées à des hématémèses, et chez qui une fibroscopie a révélé une formation battante comprimant le bulbe duodénal avec un saignement actif, une tomodensitométrie abdominale a été réalisé et a mis en évidence un anévrisme de l’artère gastroduodénale englobant son ostium, et rendant une prise en charge endovasculaire impossible. Une chirurgie ouverte a donc été réalisée, et a consisté en une mise à plat associée à une exclusion de l’anévrisme, puis complétée par une plastie du bulbe. Une tomodensitométrie post-opératoire a confirmé l’exclusion totale de l’anévrisme avec la conservation de la circulation hépatique.

Keywords: Aneurysm; gastroduodenal artery; hematemesis.

Hydrangea macrophylla (Thunb.) Ser. (Hydrangeaceae) is the most popular hydrangea species grown in home gardens and landscapes in China. Plants of H. macrophylla with symptoms of powdery mildew were found in a commercial wholesale nursery in Huzhu, Haidong (36°49'11.87" N, 101°57'03.36″E, alt. 2490 m), in May 2020, with disease incidence reaching 80%. Symptoms included yellowing and necrosis of leaves. Upon microscopic observation, masses of conidia and mycelium were observed covering the symptomatic tissues. Fungal isolates displayed nipple-shaped hyphal appressoria, often poorly developed, conidiophores erect, arising laterally or from the upper surface of hyphal mother cells, and positioned almost centrally or towards one end of the cells, up to about 160 μm long (n = 30), with foot cells straight or flexuous, 32 to 86 × 8 to 13 μm (n = 50), followed by one to three shorter cells about 11 to 24 × 10 to 15 μm (n = 50), forming catenescent conidia in usually predominantly chains, conidia doliiform to limoniform, hyaline, 24 to 35 × 13 to 25μm (n = 50). Conidial germination was of the Euoidium type. Chasmothecia were not observed. To confirm fungal classification, single spores were isolated and cultured on detached leaf bioassay following the protocol described in Farinas et al. (2019). Total DNA was extracted directly from single-spore cultures using a Chelex extraction method (Walsh et al. 1991). The rDNA internal transcribed spacer (ITS) regions were amplified by polymerase chain reaction (PCR) utilizing the universal primer pairs ITS1/ITS4 (White et al. 1990). The sequences (726-727 bp) were deposited in GenBank (accessions no. MT568633, MT757924 and MT757925). The ITS sequences showed 99.9-100% identity with a sequence of Golovinomyces orontii reported on Papaver rhoeas (AB769466) in Switzerland. Based on the ITS rDNA phylogenetic tree, the sequences retrieved from the specimen clustered within a strongly supported clade with G. orontii (AB769466), confirming the identity of the pathogen (Takamatsu 2013). Cladistic trees were constructed using the neighbor-joining method with the Kimura 2-parameter substitution model in MEGA 6.0. Branch robustness was assessed via bootstrap analysis with 1,000 replicates. To confirm pathogenicity, eight H. macrophylla plants were sprayed until run-off with a suspension containing 1 × 105 conidia/ml. Four plants were used for fulfilling Koch's postulates and four plants were used as mock-inoculated controls sprayed only with sterile distilled water. Inoculated and non-inoculated plants were covered with plastic bags separately and maintained overnight in a greenhouse at 25 ± 2°C and 50 to 60% relative humidity. Typical powdery mildew colonies developed on inoculated plants 10 to 15 days after inoculation, which were morphologically identical to those originally observed on the diseased plats, whereas the control plants remained symptomless. To our knowledge, this is the first report of powdery mildew caused by G. orontii on H. macrophylla in Qinghai-Tibet Plateau, China (Braun and Cook 2012).

Keywords: Golovinomyces orontii; Hydrangea macrophylla; Powdery Mildew; Qinghai-Tibet Plateau.

Plat castable ceramics (PCC) was bonded to SDA-I mild fusing alloy using three bonding agents, and the tensile bond strengths were tested respectively in order to select the best bonding agent and the best way of surface treatment. The results obtained with scanning electron microscope and X-ray energy spectrum showed that the bonding behavior of TF agent was the best one. The greatest tensille bond strength (24.37 MPa) was obtained with TF agent when the surface of PCC was etched by hydrofluoric acid and the alloy was painted with KH-570. Remarkable tensile bond strengths were noted in the group where EM agent was used with PCC etched by hydrofluoric acid and alloy treated with sandblasting (15.20 MPa), and in the group where Porcelite dual cure cement was applied with PCC painted by KH-570 and alloy treated with sandblasting (15.25 MPa).

Neuroblastoma is the second most common pediatric cancer involving the peripheral nervous system in which stage IVS metastatic tumors regress due to spontaneous differentiation. 13-cis retinoic acid (13-cis RA) is currently used in the clinic for its differentiation effects and although it improves outcomes, relapse is seen in half of high-risk patients. Combinatorial therapies have been shown to be more effective in oncotherapy and since cathepsin inhibition reduces tumor growth, we explored the potential of coupling 13-cis RA with a cathepsin inhibitor (K777) to enhance therapeutic efficacy against neuroblastoma. Shotgun proteomics was used to identify proteins affected by K777 and dual (13-cis RA/K777) treatment in neuroblastoma SK-N-SH cells. Cathepsin inhibition was more effective in increasing proteins involved in neuronal differentiation and neurite outgrowth than 13-cis RA alone, but the combination of both treatments enhanced the neuronal differentiation effect. SIGNIFICANCE: As neuroblastoma can spontaneously differentiate, determining which proteins are involved in differentiation can guide development of more accurate diagnostic markers and more effective treatments. In this study, we established a differentiation proteomic map of SK-N-SH cells treated with a cathepsin inhibitor (K777) and K777/13-cis RA (dual). Bioinformatic analysis revealed these treatments enhanced neuronal differentiation and axonogenesis pathways. The most affected proteins in these pathways may become valuable biomarkers of efficacy of drugs designed to enhance differentiation of neuroblastoma [1].

Keywords: ADAM10; APP; Adhesion; CRABP2; Cathepsin inhibitor; Differentiation; GDF15; Homeostasis; ICAM1; Integrin; Membrane fusion; NEFM; Neurite; Neuroblastoma; PLAT; Proteomics; RET; Retinoic acid; SQSTM1; Secretory vesicles.

African Americans (AAs) are an admixed population with widely varying proportion of West African ancestry (WAA). Here we report the correlation of WAA to gene expression and DNA methylation in AA-derived hepatocytes, a cell type important in disease and drug response. We perform mediation analysis to test whether methylation is a mediator of the effect of ancestry on expression. GTEx samples and a second cohort are used as validation. One hundred and thirty-one genes are associated with WAA (FDR < 0.10), 28 of which replicate and represent 220 GWAS phenotypes. Among PharmGKB pharmacogenes, VDR, PTGIS, ALDH1A1, CYP2C19, and P2RY1 nominally associate with WAA (p < 0.05). We find 1037 WAA-associated, differentially methylated regions (FDR < 0.05), with hypomethylated genes enriched in drug-response pathways. In conclusion, WAA contributes to variability in hepatocyte expression and DNA methylation with identified genes previously implicated for diseases disproportionately affecting AAs, including cardiovascular (PTGIS, PLAT) and renal (APOL1) disease, and drug response (CYP2C19).

In the present study, we have analyzed four highly polymorphic regions (STRs) chosen from four candidate genes involved in: (1) Platelet aggregation: alpha subunit of the platelet GpIIb/GpIIIa integrin complex (GpIIIa (CT)n; 17q21.31), (2) Coagulation fibrinolysis: Plasminogen Activator Tissue (PLAT5 (TG)14(TA)12; 8p12-q11.2) and Plasminogen Activator Inhibitor-1 (PAI-1 (CA)n; 7q21.3-q22), (3) Oxidative metabolism: the inducible nitric oxide (NO) synthase (iNOS) gene (NOS2A (CCTTT)n; 17cen-q11.2). Allele frequencies for these four STR loci were investigated in several Mediterranean populations. The population data deviate from the Hardy-Weinberg equilibrium in all populations for GpIIIa (CT)n polymorphism.

Background Nasal polyps (NP) are characterized by pseudocysts derived from stromal tissue edema and cause persistent infections in patients with chronic rhinosinusitis (CRS). A low level of tissue-type plasminogen activator (gene name PLAT) is considered a cause of stromal tissue edema because of insufficient plasmin activation in NP; however, the mechanism regulating PLAT gene expression levels is still unclear. The epigenetic mechanism regulating the PLAT gene expression has been studied in other tissues. Objective We aimed to investigate the methylation levels in the proximal PLAT promoter and their effects on gene expression in NP tissue. Methods We investigated the methylation levels at 3 CpG sites in the proximal PLAT promoter regions (-618, -121, and -105 with respect to the transcription initiation site) by bisulfite pyrosequencing and their effects on the gene expression by quantitative real-time polymerase chain reaction (qPCR) in 20 paired samples of NP and inferior turbinate tissue (IT) from patients with CRS. Results The DNA methylation levels at all CpG sites were higher ( P < .01), and the PLAT expression was lower ( P < .001) in NP compared with IT. The methylation changes at the -618 site showed a negative correlation with the gene expression changes between NP and IT ( r = -.65, P < .01). Conclusions Hypermethylation of PLAT promoter may downregulate the gene expression in NP, leading to excessive fibrin deposition by aberrant coagulation cascade. DNA methylation of proximal PLAT promoter may contribute to NP growth and have a potential as a new therapeutic target.

Factors determining the thrombogenic response to particular artificial surfaces were investigated ex vivo in a canine shunt model. Methods using radioisotopic tracers made it possible to dynamically monitor the deposition of labelled blood cells and proteins on a NHLBI.DTB primary reference material polydimethylsiloxane (PRM.PDMS) and on a IUPAC reference material polyvinyl chloride (IUPAC.PVC). On the one hand, leukocyte affinity tau s(leu) (number of deposited leukocytes mm-2s-1) was not significantly different between IUPAC.PVC (tau s(leu) = 1.2-2.5) and PRM.PDMS (tau s(leu) = 1.5-3.4) and the fibrinogen adsorption rate varied from 33 to 48.10(-5) micrograms mm-2s-1 for both these materials. On the other hand, platelet affinity tau s(plat) (number of deposited platelets mm-2s-1) was significantly different (p < 0.05) for IUPAC.PVC and PRM.PDMS (tau s(plat)PVC = 683 +/- 200>> tau s(plat)PDMS = 327 +/- 80). Scanning electron micrographs of adherent platelets, red cells and leukocytes after blood contact ex vivo were performed after each experiment. This preliminary work contributes not only to quantify the adsorption of different radiotracers, but also to evaluate the superficial distribution of the labelled biological species on the inner surface of the tested biomaterials.

OBJECTIVE

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract. About 70% of GIST occur in the stomach, 20% in the small intestine and less than 10% in the esophagus. This study aimed to explore the difference of gene expression profile of GIST between different tumor sites.

METHODS

Microarray data GSE8167 (accession number of the microarray data) were available from Gene Expression Omnibus (GEO) which included 23 gastric and 9 small intestine untreated GIST samples, and then the differentially expressed genes (DEGs) between these samples were identified using t-test. Furthermore, pathway enrichment analysis was performed to these DEGs and one protein-protein interaction network was constructed by STRING. Additionally, BioNet in R was used to establish a sub-network with false discovery rate < 0.001, and genes in this sub-network were further subjected to gene ontology (GO) and pathway analyses.

RESULTS

A total 730 genes were differentially expressed between gastric samples and small intestine samples, indicating the tissue specifity of GIST. Pathway analysis suggested these DEGs disturbed ECM-receptor interaction, gap junction and colorectal cancer. Moreover, some nodes (such as PLAT, VEGFC, PGF and CHD7) in the sub-network were significantly enriched in blood vessel development (p=4.58E-06), appendage development (p=9.54E-06) and skeletal system development (p=2.40E-04), respectively. Finally, several DEGs in the sub-network, including VEGFC and PGF, mainly affected pathways in cancer, focal adhesion, bladder cancer and cytokine-cytokine receptor interaction.

CONCLUSIONS

Our results suggest that molecular mechanisms of GIST originating in different site were different. Our findings are helpful for physicians and researchers to study the tissue specificity of GIST.

This study was aimed to investigate the signaling pathways regulating osteoclast (OC) differentiation by receptor activator of nuclear factor kappa (RANK) under physiological condition so as to provide some theoretical basis for clarifying mechanism of bone destruction in multiple myeloma. A mutant TNFR(1)/RANK(2) (named RANK-Mu) chimera consisting of tumor necrosis factor receptor 1 (TNFR(1)) and RANK intramembrane domain was constructed by using deletion mutation for deleting IVVY amino acids in RANK intramembrane domain in accordance with (535-)IVVY(-538) as specific domain regulating OC differentiation by RANK. The RANK-Mu and TNFR(1)/RANK chimera without mutation (RANK-WT) were packaged by using plat E cell line to produce the retrovirus, which were transfected into bone marrow macrophages (BMMs) of TNFR(1)/TNFR(2) double knockout mice. After stimulation of these transfected BMMs with TNF-alpha, the differentiation of BMMs into OCs were observed, meanwhile the phosphorylation of NF-kappab, JNK, p38 and ERK was detected by Western blot after stimulation of these BMMs with TNF-alpha. The results showed that BMMs transfected with RANK-WT could be differentiated into OCs and phosphorylation of NF-kappaB, JNK, P38 and ERK were activated at 5 - 10 minutes after being stimulated by TNFalpha. BMMs transfected with RANK-Mu could not be differentiated into OCs, but phosphorylation of NF-kappaB, JNK, P38 and ERK were activated also. It is concluded that RANK regulates osteoclast differentiation probably not through 4 typical signaling pathways, named as NF-kappaB, JNK, P38 and ERK, in this process other new signaling pathways maybe participate.

OBJECTIVE

To investigate the effects of Ulinastatin with different doses on pulmonary protection after cardiopulmonary bypass (CPB).

METHODS

Ninety patients after CPB were brought into this study and divided into low doses Ulinastatin group (L group, n=30, 5 000 U/kg), high doses Ulinastatin group (H group, n=30, 20 000 U/kg) and control group (C group, n= 30), respectively. When the patients were transferred into ICU after CPB, Ulinastatin was given intravenously to those in L and H group, while saline was given in C group. Blood samples were harvested at the time before the treatments (T0) and 12 hours (T1), 24 hours (T2) after the treatments, for the measurements of arterial pressure of oxygen (PaO2), arterial pressure of carbon monoxide (PaCO2),difference of alveoli-arterial oxygen pressure (PO(2A), oxygenation index (Ol),and tumor necrosis factor-alpha (TNF-alpha) level. Pulmonary dynamic compliance (Cd), plat pressure (Pplat) and peak pressure (Ppeak) were determined at the time of To and wean (Tw). The durations of ventilation and intubation were recorded.

RESULTS

At T0, the levels of PaO2, PaCO2, PO2A-a, OI and TNF-alpha in each group showed no significantly difference (P> 0. 05). At T1 and T2, the patients in H group had higher levels of PO2, PO2A-a. and OI, lower level of TNF-alpha, shorter duration of ventilation and intubation than the patients in other two groups(P<0. 05). The parameters in L group were better than those in C group, but the differences were not stastically significant (P>0. 05). There was no significantly difference in the levels of Cd, Pplat, and Ppeak at T0 and Tw between any two groups (P>0. 05). The intubation and ventilation time in H group were shorter than that in L and C group (P<0. 05).

CONCLUSIONS

The application of Ulinastatin could achieve pulmonary protective effect after CPB, and it seems the effect could be better with high dose (20 000 U/kg) of Ulinastatin.

Background: The Clinicopathological and Gene Expression Profile (CP-GEP) model was developed to accurately identify patients with T1-T3 primary cutaneous melanoma at low risk for nodal metastasis.

Objective: To validate the CP-GEP model in an independent Dutch cohort of melanoma patients.

Methods: Patients (aged ≥18 years) with primary cutaneous melanoma who underwent sentinel lymph node biopsy (SLNB) between 2007-2017 at the Erasmus Medical Centre Cancer Institute were eligible. The CP-GEP model combines clinicopathological features (age and Breslow thickness) with the expression of eight target genes involved in melanoma metastasis (ITGB3/PLAT/SERPINE2/GDF15/TGFBR1/ LOXL4/IL8/MLANA). Using the pathology result of SLNB as gold standard, performance measures of the CP-GEP model were calculated, resulting in CP-GEP high risk or low risk for nodal metastasis.

Results: In total, 210 patients were included in the study. Most patients presented with T2 (n=94,45%) or T3 (n=70,33%) melanoma. Of all patients, 27% (n=56) had a positive SLNB, with nodal metastasis in 0%, 30%, 54%, and 16% of T1, T2, T3, and T4 melanoma patients, respectively. Overall, the CP-GEP model had a negative predictive value (NPV) of 90.5% (95% confidence interval [CI]77.9-96.2), with an NPV of 100% (95%CI:72.2-100) in T1, 89.3% (95%CI:72.8-96.3) in T2, and 75.0% (95%CI:30.1-95.4) in T3 melanomas. The CP-GEP indicated high risk in all T4 melanomas.

Conclusions: The CP-GEP model is a non-invasive and validated tool that accurately identified patients with primary cutaneous melanoma at low risk for nodal metastasis. In this validation cohort, the CP-GEP model has shown potential to reduce SLNB procedures in melanoma patients.

Background & aims: It is not clear how pancreatic cancer stem cells (CSCs) are regulated, resulting in ineffective treatments for pancreatic cancer. PAF1, a RNA polymerase II-associated factor 1 complex (PAF1C) component, maintains pluripotency of stem cells, by unclear mechanisms, and is a marker of CSCs. We investigated mechanisms by which PAF1 maintains CSCs and contributes to development of pancreatic tumors.

Methods: Pancreatic cancer cell lines were engineered to knockdown PAF1 using inducible small hairpin RNAs. These cells were grown as orthotopic tumors in athymic nude mice and PAF1 knockdown was induced by administration of doxycycline in drinking water. Tumor growth and metastasis were monitored via IVIS imaging. CSCs were isolated from pancreatic cancer cell populations using flow cytometry and characterized by tumor sphere formation, tumor formation in nude mice, and expression of CSC markers. Isolated CSCs were depleted of PAF1 using the CRISPR/Cas9 system. PAF1-regulated genes in CSCs were identified via RNA-seq and PCR array analyses of cells with PAF1 knockdown. Proteins that interact with PAF1 in CSCs were identified by immunoprecipitations and mass spectrometry. We performed chromatin immunoprecipitation sequencing of CSCs to confirm the binding of the PAF1 sub-complex to target genes.

Results: Pancreatic cancer cells depleted of PAF1 formed smaller and fewer tumor spheres in culture and orthotopic tumors and metastases in mice. Isolated CSCs depleted of PAF1 downregulated markers of self-renewal (NANOG, SOX9, and BETA-CATENIN), of CSCs (CD44v6, and ALDH1), and the metastasis-associated gene signature, compared to CSCs without knockdown of PAF1. The role of PAF1 in CSC maintenance was independent of its RNA polymerase II-associated factor 1 complex component identity. We identified DDX3 and PHF5A as proteins that interact with PAF1 in CSCs and demonstrated that the PAF1-PHF5A-DDX3 sub-complex bound to the promoter region of Nanog, whose product regulates genes that control stemness. Levels of the PAF1-DDX3 and PAF1-PHF5A were increased and co-localized in human pancreatic tumor specimens, human pancreatic tumor-derived organoids, and organoids derived from tumors of KPC mice, compared with controls. Binding of DDX3 and PAF1 to the Nanog promoter, and the self-renewal capacity of CSCs, were decreased in cells incubated with the DDX3 inhibitor RK-33. CSCs depleted of PAF1 downregulated genes that regulate stem cell features (Flot2, Taz, Epcam, Erbb2, Foxp1, Abcc5, Ddr1, Muc1, Pecam1, Notch3, Aldh1a3, Foxa2, Plat, and Lif).

Conclusions: In pancreatic CSCs, PAF1 interacts with DDX3 and PHF5A to regulate expression of NANOG and other genes that regulate stemness. Knockdown of PAF1 reduces the ability of orthotopic pancreatic tumors to develop and progress in mice and their numbers of CSCs. Strategies to target the PAF1-PHF5A-DDX3 complex might be developed to slow or inhibit progression of pancreatic cancer.

Keywords: mouse model; pancreatic ductal adenocarcinoma; side population; transcription.

Tissue plasminogen activator (tPA) has been shown to prevent steroid-induced reduction in aqueous humor outflow facility via an upregulation in matrix metalloproteinase (Mmp) expression. The purpose of this study was to determine whether tPA can rescue outflow facility reduction in the Tg-MYOC^Y437H mouse model, which replicates human juvenile open angle glaucoma. Outflow facility was measured in Tg-MYOC^Y437H mice following: periocular steroid exposure and intraocular protein treatment with enzymatically active or enzymatically inactive tPA. Effects of tPA on outflow facility were compared to those of animals treated with topical sodium phenylbutarate (PBA), a modulator of endoplasmic reticulum stress. Gene expression of fibrinolytic pathway components (Plat, Plau, and Pai-1) and matrix metalloproteinases (Mmp-2, -9, and -13) was determined in angle ring tissues containing the trabecular meshwork. Tg-MYOC^Y437H mice did not display further outflow facility reduction following steroid exposure. Enzymatically active and enzymatically inactive tPA were equally effective in attenuating outflow facility reduction in Tg-MYOC^Y437H mice and caused enhanced expression of matrix metalloproteinases (Mmp-9 and Mmp-13). tPA was equally effective to topical PBA treatment in ameliorating outflow facility reduction in Tg-MYOC^Y437H mice. Both treatments were associated with an upregulation in Mmp-9 expression while tPA also upregulated Mmp-13 expression. tPA increases the expression of matrix metalloproteinases and may cause extracellular matrix remodeling at the trabecular meshwork, which results in reversal of outflow facility reduction in Tg-MYOC^Y437H mice.

Keywords: Matrix metalloproteinase (MMP); Myocilin; Open angle glaucoma; Outflow facility; Tissue plasminogen activator (tPA); Trabecular meshwork.

A pharmacokinetic study on cis-(glycolato-O,O)/ (4R,5R)-4,5-bis(aminomethyl)-1,3-dioxolane-2-spiro-1'- cyclohexane]platinum(II) (SKI 2032R) was performed in dogs. A single dose of 2.0 mg/kg of SKI 2032R was administered i.v. bolus to three beagle dogs. Plasma samples were analyzed for platinum by flameless atomic absorption spectrophotometry. Plasma concentrations of total and ultrafiltrable platinum for SKI 2032R declined in a biexponential fashion. The mean area under the concentration-time curve (AUC0 ->> infinity) determined for ultrafiltrable platinum derived from SKI 2032R, as an active component, was 2.36 +/- 0.23 micrograms. h/ml (mean +/- S.D.), with an initial half-life of 0.23 +/- 0.20 hour, a terminal half-life of 1.32 +/- 0.49 hour, a total clearance of 14.17 +/- 1.50 ml/min/kg, and a steady-state volume of distribution of 1.21 +/- 0.24 l/kg.

Using technology to track endemic areas of communicable diseases is possible nowadays. Effectual use of such facilities, especially in developing countries, will increase earlier detection of cases as well as aid in the formulation of effective prevention strategies.

A retrospective data analysis was carried out by collecting the details of patients presented with positive dengue serology, during the outbreak season in the second half of 2012, at Kovai Medical Center and Hospital, Coimbatore, India. Clinical variables were analysed statistically using SPSS 20 and geographical mapping of the cases was carried out using EPI INFO 7 software.

1004 dengue positive cases were identified during the study period. Geographical mapping of the case clusters showed specific areas in the city as well as neighbouring districts, which were an indirect evidence of the causative mosquito's endemic breeding places. Overall mortality noted in this group was 1.3% and mortality in cases with severe thrombocytopenia was 4 in 1000 cases. Severe thrombocytopenia (Plat≤ 10,000) on admission increased odds ratio for mortality i.e. around 10 times higher than the rest of the cohorts.

Identification of endemic mosquito breeding places and implementation of proper preventive measures is always a crucial step in the prevention of further outbreaks. Effective registry using softwares by tertiary care hospitals will be obligatory to track the location of the cases as these hospitals are the nodal point of care for most of the cases in developing countries.

There is an urgent demand for affordable, rapid and easy-to-use technology to simultaneously detect many different DNA targets within one reaction. Conventional multiplex PCR is an effective methodology to simultaneously amplify different DNA targets. However, its multiplicity is limited due to the intrinsic interference and competition among primer pairs within one tube. Here, we present an easy multiplex PCR microchip system, which can simultaneously detect 54 targets. The design of the microchip is quite simple. There is a microchannel connected with multiple underlying parallel microwells. And every microchannel has an inlet/outlet for loading PCRmix. The surface of the microchannel is hydrophobic and the inner surface of the microwell is hydrophilic, which enables us to load and separate the PCRmix into different microwells simultaneously. Different primer pairs and low melting agarose are pre-fixed in different microwells, and the microchip is assembled with top glass. The PCRmix is loaded into inlets and then mineral oil is sequentially pipetted into channels to push the PCRmix into all microwells and subsequently mineral oil fills the channels to avoid cross contaminations. After the PCRmix is loaded, it would be placed on a plat thermal cycler for PCR. During PCR, the low melting gel in the well is liquid and after PCR it would be solidified due to temperature changes. When PCR is completed, a nucleic acid dye is introduced into channels and then results are visualized by a home-made, potable UV detector. In our platform we successfully detected seven frequently used targets of genetically modified (GM) organisms. The results demonstrate that our platform has high flexibility and specificity. Due to the excellent performance of this technology, we believe that it can be applied to multiple nucleic acid detection fields including GM organisms.

It is well-known that indomethacin (the cyclooxygenase 1 & 2 inhibitor) and RU486 (or mifepristone, the progesterone receptor antagonist) block follicular rupture in rats. To characterize genetic alterations in unruptured follicles, gene expression profiles in ovarian follicle were analyzed in indomethacin- and RU486-treated female Sprague-Dawley rats. Ovaries are collected at 22:00 on the proestrus day and 10:00 on the following estrus day after a single dose of indomethacin and RU486. Histopathologically, changes depicting responses to LH surge were observed in ovaries, uteri and vagina. Total RNA was extracted from pre-ovulatory follicles or unruptured follicles collected by laser microdissection and analyzed by Genechip(®). Among genes showing statistically significant changes compared to control groups, following changes were considered relevant to induction of unruptured follicles. In indomethacin-treated rats, Wnt4 was down-regulated, suggesting effect on tissue integrity and steroid genesis. In RU486-treated rats, Adamts1, Adamts9, Edn2, Ednra, Lyve1, Plat, and Pparg were down-regulated. These changes suggest effects on proteolysis for extra cellular matrix or surrounding tissue (Adamts1 & 9, and Plat), constriction of smooth muscle surrounding follicles (Edn2, Ednra, and Pparg), follicular fluid (Lyve1), and angiogenesis (Pparg). Down-regulation of angiogenesis related genes (Angpt2, Hmox1, and Vegfa) was observed in both treatment groups. Here, we clarify genetic alterations induced by the inhibition of cyclooxygenase or progesterone receptor.

To investigate the changes in the concentrations of interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and 8-iso-prostaglandin F2α (8-isoPGF2α) in exhaled breath condensate (EBC) in patients undergoing cardiac valve replacement under cardiopulmonary bypass (CPB) and its relationship with postoperative acute respiratory distress syndrome (ARDS).A prospective, case-control study was performed on 55 patients undergoing elective cardiac valve replacement under cardiopulmonary bypass, between November 2017 and May 2019. According to the diagnosis of postoperative ARDS, the patients were divided into ARDS group and control group. We compared the clinical characteristics, outcomes, respiratory mechanics, oxygenation parameters, and mediators in the 2 groups immediately after tracheal intubation (T1), at the end of CPB (T2), and 2 hours (T3) and 6 hours (T4) after CPB, and calculated the receiver operating characteristic curve (ROC), sensitivity, and specificity of the corresponding mediators.ARDS occurred in 29 patients after CPB. The ARDS group exhibited prolonged postoperative ventilator support, time to extubation, length of stay in the ICU, and postoperative length of stay. The peak airway pressure (Ppeak) and plat airway pressure (Pplat) at T4 were higher in the ARDS group compared with the control group. The alveolar-arterial oxygen partial pressure [P(A-a)O2] and respiratory index (RI) were higher and PaO2/FiO2 was lower in the ARDS group at T2-4 compared with the control group. The levels of EBC and serum mediators in the ARDS group were significantly higher at T2-4 compared with those in the control group. All the mediators in EBC were correlated significantly with those in the serum in the ARDS group (r = 0.7314, 0.898, 0.8386, 0.792) and control group (r = 0.6093, 0.8524, r = 0.7828, r = 0.6575) (P < .001). Meanwhile, the area under the curve (AUC) of IL-8 in EBC was significantly lower at T2 and the AUC of IL-6 in EBC was significantly higher at T4 than in serum (P < .05). In addition, all of the mediators in EBC had a certain accuracy in diagnose of postoperative ARDS.EBC analysis could be used to predict the high incidence of ARDS after cardiac valve replacement under CPB.

Recent literature suggests that optimization of tidal driving pressure (ΔP) would be a better variable to target for lung protection at the bedside than tidal volume (V_T) or plateau pressure (P_plat), the traditional indicators of ventilator-induced lung injury. However, the usual range or variability of ΔP over time for any subject category have not been defined. This study sought to document the ΔP ranges observed in current practice among mechanically ventilated subjects receiving routine care for diverse acute conditions in a community hospital environment.

This was a retrospective, observational study in a university-affiliated and house staff-aided institution with respiratory care protocols based on extant lung-protective guidelines for V_T. Demographic characteristics and measured parameters related to mechanical ventilation and hemodynamics were extracted from electronic records of intubated subjects for each 8-h period of the first 24 h in the ICU. P_plat values reported by the ventilator were validated by the respiratory therapist before those data were entered into the electronic medical record.

The mean ΔP was significantly higher at Time 1 (mean 16.1, range 7.0-31.0 cm H₂O) compared to both Time 2 (mean 14.5, range 7.0-35.0 cm H₂O) (P < .001) and Time 3 (mean 14.8, range 8.0-32 cm H₂O) (P < .001). At all time points, the median ΔP was higher for completely passive breathing compared to triggered breathing. The widest difference between presumed entirely passive and presumed intermittently or consistently triggered breaths occurred at Time 1 (mean ΔP = 17.2 vs 14.9 cm H₂O, respectively) (P = .01).

Suggested safety thresholds for ΔP are often violated by a strategy that focuses on only V_T and P_plat. Our data suggest that ΔP is lower for passive versus triggered breathing cycles. Vigilance is especially important in the initial stages of mechanical ventilator support, and attention should be paid to triggering efforts when interpreting and comparing machine-determined numerical values for ΔP.

In North Carolina (NC), biosolids land application rates governed by crop nitrogen (N) requirements typically surpass crop phosphorus (P) needs, increasing surface water pollution potential. The NC Department of Environmental Quality (NCDEQ) is considering P-based biosolids application guidelines for some nutrient-impaired watersheds using the P Loss Assessment Tool (PLAT), but important biosolids information is lacking: total P (TP), water-extractable P (WEP), and percent water-extractable P (PWEP). In each of three seasons, we sampled 28 biosolids from 26 participating water resource recovery facilities (WRRFs) and analyzed for TP, WEP, and percent dry matter (DM), from which PWEP and nonsoluble P were calculated. Based on descriptive statistics and an online survey of treatment processes, biosolids were divided into Class A-alkaline, Class A-heat, Class B-slurry, and Class B-cake. The average TP in Class A alkaline stabilized biosolids was more than five times less than the average of the other biosolids, 5.0 vs. 26.6 g/kg, respectively. Averaged over biosolids, WEP and PWEP were 1.4 g/kg and 5.0%, respectively. Stabilization processes appeared to reduce WEP substantially, so biosolids potential soluble-P loss is low. Our data will allow PLAT to be used for biosolids P-loss risk assessments.

Objective: To investigate the effect of pulmonary vascular dysfunction in the prognosis of patients with acute lung injury (ALI).

Methods: Patients with ALI who underwent pulmonary artery catheterization in the department of critical care medicine of Wuhan NO.1 Hospital from June 2017 to June 2019 were enrolled. The general information, clinical and hemodynamic indexes [central venous pressure (CVP), pulmonary artery wedge pressure (PAWP), pulmonary artery systolic pressure (sPAP), pulmonary artery diastolic pressure (dPAP), mean pulmonary artery pressure (mPAP), cardiac index (CI)], acute physiology and chronic health evaluation II (APACHE II) score, arterial blood gas parameters [pH, partial pressure of oxygen (PO₂), partial pressure of carbon dioxide (PCO₂), oxygenation index (PaO₂/FiO₂)], whether there was shock or not; ventilator parameters [platform pressure (Plat), positive end-expiratory pressure (PEEP)], etc. were recorded. Pulmonary artery oxygen saturation, pulmonary vascular function indexes [transpulmonary potential gradient (TPG) and pulmonary vascular resistance index (PVRi)] were calculated. The relationship between TPG, PVRi and mechanical ventilation time, the length of intensive care unit (ICU) stay, cardiovascular days and 60-day mortality were analyzed in patients with different prognosis of 60-day and whether the TPG increased (≥ 12 mmHg was defined as elevated TPG, 1 mmHg = 0.133 kPa).

Results: A total of 65 patients were included in the study, including 30 males and 35 females; aged (48.9±15.2) years old. Forty-eight cases survived in 60-days, 17 died, and the 60-day mortality was 26.2%. At the baseline, there were no significant differences in cardiopulmonary function measurements, such as CVP, sPAP, dPAP, PAWP, CI, etc. between the two groups of patients with different prognosis. The APACHE II score, shock ratio, TPG and PVRi of the death group were significant higher than those of the survival group [APACHE II: 34±9 vs. 28±11, shock: 52.9% vs. 25.0%, TPG (mmHg): 16.2±1.9 vs. 14.6±2.1, PVRi (kPa×s×L^-1): 31.8±4.2 vs. 29.7±3.5, all P < 0.05]. The 60-day mortality of 47 patients with TPG ≥ 12 mmHg was significantly higher than that of 18 patients with TPG < 12 mmHg (34.0% vs. 5.6%), and the mechanical ventilation time and the length of ICU stay were also significantly longer (days: 17±9 vs. 11±8, 16±5 vs. 12±5), and the cardiovascular days also increased significantly (days: 23±7 vs. 18±6), and the differences were statistically significant (all P < 0.05). Pearson correlation analysis showed that PVRi was significantly correlated with mechanical ventilation time, the length of ICU stay and cardiovascular days (r₁ = 0.317, P₁ = 0.030; r₂ = 0.277, P₂ = 0.005; r₃ = 0.285, P₃ = 0.002). In the individual multivariate Logistic regression model, the highest PVRi was an independent risk factor for the 60-day mortality [odds ratio (OR) = 30.5, 95% confidence interval was 20.4-43.1, P = 0.023].

Conclusions: Pulmonary vascular dysfunction is common in ALI patients and is independently associated with adverse outcomes.