In this study, enzyme-linked immunosorbent assay has been used to examine the frequencies of serum autoantibodies against two candidate tumor-associated antigens intensively selected from the Human Protein Atlas database, in combination with 13 tumor-associated antigens available from our lab in sera from 44 OC patients and 50 normal healthy controls. Conventional evaluation (mean + 3SD as the cutoff value to determine a positive reactivity), receiver operating characteristic curve analyses, and classification tree analysis were further used to evaluate the diagnostic performance of autoantibodies against these tumor-associated antigens (anti-tumor-associated antigens) in ovarian cancer. For single anti-tumor-associated antigen, when the cutoff values were set as mean + 3SD of normal healthy controls, NPM1, MDM2, PLAT, p53, and c-Myc could achieve sensitivity higher than 20% at 98% specificity. Combinational utilization of autoantibodies against MDM2, PLAT, NPM1, 14-3-3 Zeta, p53, and RalA achieved the optimal diagnostic performance with 72.7% sensitivity at 96% specificity. Receiver operating characteristic curve analysis showed that the area under the receiver operating characteristic curves of autoantibodies against c-Myc, NPM1, MDM2, p16, p53, and 14-3-3 Zeta were greater than 0.80. This indicated that these tumor-associated antigens held high potential to serve as diagnostic biomarkers in ovarian cancer detection. Decision tree analysis indicated that anti-c-Myc held high potential in the detection of ovarian cancer. Further studies are warranted to validate the diagnostic performance of these anti-tumor-associated antigens with high area under the receiver operating characteristic curve, including autoantibodies against c-Myc, MDM2, PLAT, NPM1, 14-3-3 Zeta, p53, and RalA.

BACKGROUND

The long-term molecular changes in the central nervous system constitute an important aspect of general anaesthesia, but little is known about to what extent these molecular changes are affected by anaesthesia duration. The aim of the present study was to evaluate the effects of short duration (20 min) general anaesthesia with isoflurane or avertin on the expression of 20 selected genes in the mouse hippocampus at 1 and 4 days after anaesthesia.

METHODS

Nine to eleven-weeks-old male mice received one of the following treatments: 20 min of avertin-induced anaesthesia (n=11), 20 min of isoflurane-induced anaesthesia (n=10) and no anaesthesia (n=5). One and four days after anaesthesia, gene expression in the hippocampus was determined with reverse transcription quantitative real-time polymerase chain reaction.

RESULTS

We found that anaesthesia led to the upregulation of six genes: Hspd1 (heat shock protein 1), Plat (tissue plasminogen activator) and Npr3 (natriuretic peptide receptor 3) were upregulated only 1 day after anaesthesia, whereas Thbs4 (thrombospondin 4) was upregulated only 4 days after anaesthesia. Syp (synaptophysin) and Mgst1 (microsomal glutathione S-transferase 1) were upregulated at both time points. Hspd1, Mgst1 and Syp expression was increased regardless of the anaesthetic used, Npr3 and Plat were increased only in mice exposed to avertin, and Thbs4 was upregulated only after isoflurane-induced anaesthesia.

CONCLUSIONS

This study shows that some of the effects of short general anaesthesia on gene expression in the mouse hippocampus persist for at least 4 days.

Understanding the host regulatory mechanisms opposing virus infection and virulence can provide actionable insights to identify novel therapeutics against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We have used a network biology approach to elucidate the crucial factors involved in host responses involving host-microRNA (miRNA) interactions with host and virus genes using recently published experimentally verified protein-protein interaction data. We were able to identify 311 host genes to be potentially targetable by 2,197 human miRNAs. These miRNAs are known to be involved in various biological processes, such as T-cell differentiation and activation, virus replication, and immune system. Among these, the anti-viral activity of 38 miRNAs to target 148 host genes is experimentally validated. Six anti-viral miRNAs, namely, hsa-miR-1-3p, hsa-miR-17-5p, hsa-miR-199a-3p, hsa-miR-429, hsa-miR-15a-5p, and hsa-miR-20a-5p, are previously reported to be anti-viral in respiratory diseases and were found to be downregulated. The interaction network of the 2,197 human miRNAs and interacting transcription factors (TFs) enabled the identification of 51 miRNAs to interact with 77 TFs inducing activation or repression and affecting gene expression of linked genes. Further, from the gene regulatory network analysis, the top five hub genes HMOX1, DNMT1, PLAT, GDF1, and ITGB1 are found to be involved in interferon (IFN)-α2b induction, epigenetic modification, and modulation of anti-viral activity. The comparative miRNAs target identification analysis in other respiratory viruses revealed the presence of 98 unique host miRNAs targeting SARS-CoV-2 genome. Our findings identify prioritized key regulatory interactions that include miRNAs and TFs that provide opportunities for the identification of novel drug targets and development of anti-viral drugs.

Keywords: SARS-CoV-2; TFS; hub genes; miRNA; regulatory network.

Luminol electrochemiluminescence (ECL) imaging was developed for the parallel measurement of active membrane cholesterol at single living cells, thus establishing a novel electrochemical detection technique for single cells with high analysis throughput and low detection limit. In our strategy, the luminescence generated from luminol and hydrogen peroxide upon the potential was recorded in one image so that hydrogen peroxide at the surface of multiple cells could be simultaneously analyzed. Compared with the classic microelectrode array for the parallel single-cell analysis, the plat electrode only was needed in our ECL imaging, avoiding the complexity of electrode fabrication. The optimized ECL imaging system showed that hydrogen peroxide as low as 10 μM was visible and the efflux of hydrogen peroxide from cells could be determined. Coupled with the reaction between active membrane cholesterol and cholesterol oxidase to generate hydrogen peroxide, active membrane cholesterol at cells on the electrode was analyzed at single-cell level. The luminescence intensity was correlated with the amount of active membrane cholesterol, validating our system for single-cell cholesterol analysis. The relative high standard deviation on the luminescence suggested high cellular heterogeneities on hydrogen peroxide efflux and active membrane cholesterol, which exhibited the significance of single-cell analysis. This success in ECL imaging for single-cell analysis opens a new field in the parallel measurement of surface molecules at single cells.

Hypertrophic chondrocytes exist in two forms detectable by electron microscopy, light and dark chondrocytes; the functional implications of the heterogeneous morphology are unknown. The aims of the study were to establish a method for separating light from dark hypertrophic chondrocytes and to identify genes differentially expressed between the two populations. Three-dimensional pellet cultures of chondrocytes from cartilage of neonatal rats were induced to undergo hypertrophy by treatment with triiodothyronine. Cultures were dissociated and subjected to density gradient centrifugation. The cell fraction with the lowest density comprised predominantly light hypertrophic chondrocytes, and the fraction with the highest density comprised predominantly dark hypertrophic chondrocytes. An Affymetrix GeneChip rat expression array was used to compare expression between dark cell-containing pellets and the light cell-enriched fraction. Genes identified on the array as putative dark cell-selective genes included genes encoding extracellular matrix proteins and enzymic modulators thereof. Expression of a subset of genes (Col1a1, periostin, osteoglycin, tPA/Plat, and Chst11) was confirmed as dark cell-selective using quantitative reverse transcriptase polymerase chain reaction. The most highly differentially expressed dark cell-selective gene was periostin. In immunocytochemical studies of light and dark cell-enriched fractions, periostin staining was detectable in dark, but not light hypertrophic chondrocytes. The results provide insight into molecular differences between light and dark hypertrophic chondrocytes.

The two-spotted spider mite Tetranychus urticae is an important pest with an exceptionally broad host plant range. This generalist rapidly acclimatizes and adapts to a new host, hereby overcoming nutritional challenges and a novel pallet of constitutive and induced plant defenses. Although recent studies reveal that a broad transcriptomic response upon host plant transfer is associated with a generalist life style in arthropod herbivores, it remains uncertain to what extent these transcriptional changes are general stress responses or host-specific. In the present study, we analyzed and compared the transcriptomic changes that occur in a single T. urticae population upon long-term transfer from Phaseolus vulgaris to a similar, but chemically defended, host (cyanogenic Phaseolus lunatus) and to multiple economically important crops (Glycine max, Gossypium hirsutum, Solanum lycopersicum and Zea mays). These long-term host plant transfers were associated with distinct transcriptomic responses with only a limited overlap in both specificity and directionality, suggestive of a fine-tuned transcriptional plasticity. Nonetheless, analysis at the gene family level uncovered overlapping functional processes, recruiting genes from both well-known and newly discovered detoxification families. Of note, our analyses highlighted a possible detoxification role for Tetranychus-specific short-chain dehydrogenases and single PLAT domain proteins, and manual genome annotation showed that both families are expanded in T. urticae Our results shed new light on the molecular mechanisms underlying the remarkable adaptive potential for host plant use of generalist arthropods and set the stage for functional validation of important players in T. urticae detoxification of plant secondary metabolites.

Background: With recent advances in technology, patients with acute respiratory distress syndrome (ARDS) and severe acute exacerbations of chronic obstructive pulmonary disease (ae-COPD) could benefit from extracorporeal CO₂ removal (ECCO₂R). However, current evidence in these indications is limited. A European ECCO₂R Expert Round Table Meeting was convened to further explore the potential for this treatment approach.

Methods: A modified Delphi-based method was used to collate European experts' views to better understand how ECCO₂R therapy is applied, identify how patients are selected and how treatment decisions are made, as well as to identify any points of consensus.

Results: Fourteen participants were selected based on known clinical expertise in critical care and in providing respiratory support with ECCO₂R or extracorporeal membrane oxygenation. ARDS was considered the primary indication for ECCO₂R therapy (n = 7), while 3 participants considered ae-COPD the primary indication. The group agreed that the primary treatment goal of ECCO₂R therapy in patients with ARDS was to apply ultra-protective lung ventilation via managing CO₂ levels. Driving pressure (≥ 14 cmH₂O) followed by plateau pressure (P_plat; ≥ 25 cmH₂O) was considered the most important criteria for ECCO₂R initiation. Key treatment targets for patients with ARDS undergoing ECCO₂R included pH (> 7.30), respiratory rate (< 25 or < 20 breaths/min), driving pressure (< 14 cmH₂O) and P_plat (< 25 cmH₂O). In ae-COPD, there was consensus that, in patients at risk of non-invasive ventilation (NIV) failure, no decrease in PaCO₂ and no decrease in respiratory rate were key criteria for initiating ECCO₂R therapy. Key treatment targets in ae-COPD were patient comfort, pH (> 7.30-7.35), respiratory rate (< 20-25 breaths/min), decrease of PaCO₂ (by 10-20%), weaning from NIV, decrease in HCO₃^- and maintaining haemodynamic stability. Consensus was reached on weaning protocols for both indications. Anticoagulation with intravenous unfractionated heparin was the strategy preferred by the group.

Conclusions: Insights from this group of experienced physicians suggest that ECCO₂R therapy may be an effective supportive treatment for adults with ARDS or ae-COPD. Further evidence from randomised clinical trials and/or high-quality prospective studies is needed to better guide decision making.

Keywords: Acute respiratory distress syndrome; CO2 removal; Chronic obstructive pulmonary disease; Consensus; Driving pressure; ECCO2R; Gas exchange; Lung protective ventilation; Therapy experience; Tidal volume.

BACKGROUND

Recently, genome-wide association studies identified a pleiotropic gene locus, ABO, as being significantly associated with hematological traits. To confirm the effects of ABO on hematological traits, we examined the link between the ABO locus and hematological traits in Korean population-based cohorts.

RESULTS

Six tagging SNPs for ABO were analyzed with regard to their effects on hematological traits [white blood cell count (WBC), red blood cell count (RBC), platelet (Plat), mean corpuscular volume (MCV), and mean corpuscular haemoglobin concentration (MCHC)]. Linear regression analyses were performed, controlling for recruitment center, sex, and age as covariates. Of the 6 tagging SNPs, 3 (rs2073823, rs8176720, and rs495828) and 3 (rs2073823, rs8176717, and rs687289) were significantly associated with RBC and MCV, respectively (Bonferroni correction p-value criteria < 0.05/6 = 0.008). rs2073823 and a reported SNP (rs8176746), as well as rs495828 and a reported SNP (rs651007), showed perfect linkage disequilibrium status (r2s = 0.99). Of the remaining 3 SNPs (rs8176720, rs8176717 and rs687289), rs8176717 generated an independent signal with moderate p-value (= 0.045) when it was adjusted for by rs2073823 (the most significant SNP). We also identified a copy number variation (CNV) that was tagged by the SNP rs8176717, the minor allele of which correlated with the deletion allele of CNV. Our haplotype analysis indicated that the haplotype that contained the CNV deletion was significantly associated with MCV (β ± se = 0.363 ± 0.118, p =2.09 × 10-3).

CONCLUSIONS

Our findings confirm that ABO is one of the genetic factors that are associated with hematological traits in the Korean population. This result is notable, because GWASs fail to evaluate the link between a CNV and phenotype traits.

Bone marrow multipotent (CFU-Mix) and unipotent (CFU-GM and BFU-E) progenitor cells in the donor marrow inoculums were measured in 24 histocompatible sibling bone marrow transplants. The number of donor marrow nucleated cells, CFU-Mix, CFU-GM and BFU-E given per kilogram (kg) of recipient's body weight were 2.4 +/- 0.6 X 10(8), 3.6 +/- 4.2 X 10(3), 4.9 +/- 3.3 X 10(-4) and 4.3 +/- 4.1 X 10(4) respectively (mean +/- S.D.). Fast engraftment patients, as assessed by rise in peripheral blood neutrophils (greater than or equal to 0.5 and greater than or equal to 1.0 X 10(9)/l) and platelets (greater than 20 and greater than 50 X 10(9)/l), received a significantly greater amount of CFU-Mix/kg (greater than 3 X 10(3)/kg, p less than 0.025) and CFU-GM/kg (greater than 3 X 10(4)/kg, p less than 0.05 except for plat greater than or equal to 20 X 10(9)/l) than the slow recovery patients. Significant correlations were found between the donor CFU-Mix/kg infused and neutrophil recovery to 1 X 10(9)/l and platelet to 50 X 10(9)/l (Spearman's rank correlation coefficient r = 0.38, p = 0.04 and r = 0.58, p = 0.003, respectively). The amount of donor CFU-GM/kg given also correlated significantly to neutrophil (1 X 10(9)/l) and platelet (50 X 10(9)/l) recovery, (r = 0.33 and r = 0.37, respectively, p less than or equal to 0.05). There was no association between BFU-E, and marrow nucleated cells infused per kg and haemopoietic recovery. A number of clinical parameters were also examined to determine other factors that may influence the rate of engraftment. Acute graft vs host disease (greater than or equal to grade II) and methotrexate therapy post-transplant delayed the platelet regeneration. The results of the present report indicate that in vitro measurement of donor CFU-Mix and CFU-GM progenitors infused, correlate with the speed of granulocyte and platelet recovery in clinical allogeneic bone marrow transplants.

High-dose chemotherapy (HDCT) and autologous bone-marrow transplantation (ABMT) are widely used in the salvage treatment of non-seminomatous germ cell tumours (NSGCT). We compiled 10 published series with NSGCT patients treated by HDCT and ABMT. Several prognostic factors for long-term non-evolutive disease (NED) were studied: dose of etoposide (ETO), oxazaphosphorine derivative (OXA) (expressed in cyclophosphamide equivalents using a cyclophosphamide/ifosfamide ratio of 1:3), platin-derivate (PLAT) (expressed in cisplatin equivalents using a cisplatin/carboplatin ratio of 1:4), disease status (refractory or responder), OXA and PLAT compounds. Strong interactions were shown between disease status and PLAT and ETO. In refractory patients, logistic regression analysis showed that the doses of OXA and PLAT increase the probability of NED. Conversely, in responder patients only ETO and OXA dosages increase the probability of NED. It is concluded that the status of the disease is the most important prognostic factor for long-term NED after HDCT + ABMT in NSGCT.

The full-length cDNA of the gene PoLOX1 encoding a lipoxygenase (LOX) and its corresponding genomic DNA were isolated from the basidiomycete mushroom Pleurotus ostreatus strain H1. The deduced amino acid sequence of PoLOX1 showed similarity to a valencene dioxygenase of Pleurotus sapidus, putative LOX-like proteins from ascomycete, basidiomycete, and deuteromycete fungi, and known LOXs from plants, animals, and bacteria. PoLOX1 also contained the LOX iron-binding catalytic domain in the C-terminal region, but not the polycystin-1, lipoxygenase, alpha-toxin (PLAT) domain, which is usually found in the N-terminal region of eukaryotic LOXs. Genomic sequence analysis revealed that PoLOX1 was interrupted by one intron, and that the promoter region included TATA and CAAT boxes. Southern blot analysis indicated that PoLOX1 is a member of a small gene family comprising highly similar genes. Northern blot analysis revealed that it is transcribed more abundantly in the stipes of the fruit bodies than in the caps.

Plant growth and consequently crop yield can be severely compromised by abiotic and biotic stress conditions. Transgenic approaches that resulted in increased tolerance against abiotic stresses often were typically accompanied by adverse effects on plant growth and fitness under optimal growing conditions. Proteins that belong to the <em>PLAT</em>-plant-stress protein family harbour a single <em>PLAT</em> (Polycystin, Lipoxygenase, Alpha-toxin and Triacylglycerol lipase) domain and are ubiquitously present in monocot and dicot plant species. Until now, only limited data is available for <em>PLAT</em>-plant-stress family members, which suggested that these proteins in general could promote tolerance towards stress responses. We studied the function of the Arabidopsis <em>PLAT</em>-plant-stress protein At<em>PLAT</em>1 employing heterologous gain-of-function analysis in tobacco. At<em>PLAT</em>1 conferred increased abiotic stress tolerance in tobacco, evident by improved tolerance towards cold, drought and salt stresses, and promoted growth, reflected by a faster development under non-stressed conditions. However, the overexpression of At<em>PLAT</em>1 in tobacco reduced the tolerance towards biotic stress conditions and, therefore, could be involved in regulating the crosstalk between abiotic and biotic stress responses. Thus, we showed that heterologously expressed At<em>PLAT</em>1 functions as positive regulator of abiotic stress tolerance and plant growth, which could be an important new asset for strategies to develop plants with improved abiotic stress tolerance, without growth and subsequent yield penalties under optimal growth conditions.

The cytoplasm of laticifers, which are plant cells specialized for rubber production and defense against microbes and herbivores, is a latex. Although laticifers share common functions, the protein constituents of latexes are highly variable among plant species and even among organs. In this study, transcriptomic and proteomic analyses of Euphorbia tirucalli's (Euphorbiaceae) latex were conducted to determine the molecular basis of the laticifer's functions in this plant. The hybrid de novo assembly of Illumina mRNA-seq and expressed sequence tags obtained by Sanger's sequencing revealed 26,447 unigenes. A unigene similar to Arabidopsis embryo-specific protein 3 (AT5G62200), which is a PLAT domain-containing protein, and rubber elongation factor showed the highest expression levels. The proteome analysis, studied by liquid chromatography-mass spectrometry with the de novo assembled unigenes as the database, revealed 161 proteins in the latex, 107 of which were not detected in the stem. A gene ontology analysis indicated that the laticifer's proteome was enriched with proteins related to proteolysis, phosphatase, defense against various environmental stresses and lipid metabolisms. D-mannose-binding lectin, ricin (which lacked the N-terminal conserved ribosome-inactivating protein domain), chitinase and peroxidase were highly accumulated, as confirmed by two-dimensional polyacrylamide gel electrophoresis. Thus, the lectins and chitinase may be the major defensive proteins against pests, and the other defense-related proteins and transcripts detected in latex may work in coordination with them. Highly expressing unigenes with unknown functions are candidate novel defense- or rubber production-related genes.

OBJECTIVE

To report two novel LOXHD1 mutations, including missense mutations and the clinical features of the patients.

METHODS

We studied a three-generation Japanese family with hearing loss. Targeted next-generation sequencing was used for genetic analysis. Conditional orientation response audiometry and pure tone audiometry were used to assess hearing. SWISS-MODEL was used for molecular modeling of the PLAT domain in LOXHD1 protein.

RESULTS

The two sisters, who had either mild or severe high-frequency hearing loss, were compound heterozygous for two novel mutations (c.5674G>T [p.V1892F] and c.4212+1G>A) in LOXHD1, which is responsible for autosomal-recessive nonsyndromic hearing loss DFNB77. These cases showed less severe hearing impairment than the previously reported cases carrying LOXHD1 mutations, but their hearing loss appeared to be progressive. Molecular modeling predicted that distorted structure of the PLAT domain in the p.V1892F mutant could lead to decreased affinity of the protein to lipid membrane resulting in hair cell dysfunction.

CONCLUSIONS

We report a Japanese family carrying compound heterozygotes of truncating and nontruncating mutations in LOXHD1 identified by targeted NGS analysis. The fact of lower degree of hearing impairment in our cases than previously reported and the molecular modeling of the missense mutant provide insight to the genotype-phenotype correlation of DFNB77.

BACKGROUND

Kallmann syndrome is a disease clinically characterized by the association of hypogonadotrophic hypogonadism and anosmia or hyposmia. Most cases have been recorded among men. It is a genetic disorder with a specific gene location on the X chromosome. The cells that normally express luteinizing hormone-releasing hormone or LHRH fail to migrate the olfactory placode to the forebrain. The lateral projections of the olfactory placode also fail to induce development of the olfactory bulbs and tracts.

METHODS

The aim of this study was to compare the MRI appearance of the olfactory sulci, the olfactory bulbs and frontal lobe between groups. The first reference group was composed of 20 subjects and the second group of 18 patients suffering from Kallmann syndrome. For all studies we used a 1.5 T magnet system (Signa GE). We performed two sagittal and coronal T1-weighted sequences in spin echo (TR = 600 ms, TE = 12 ms) with interleaved 3 mm slices and a 14 cm field of view.

RESULTS

In the first group, the two olfactory bulbs were always seen on coronal slices just behind the crista galli measuring 2 to 3.2 mm transversally. On sagittal slices, in 60% of the cases two bulbs were seen (3 mm laterally of the pituitary stalk) and in the other 40% only one bulb was seen. The length of the bulb has been measured between 6 and 11 mm. We noticed a plat frontal lobe in 85% of the cases. In the second group the olfactory bulbs were never visible among the 18 patients suffering from Kallmann syndrome. The hypoplasic sulci were hardly visible and their size was less or equal to 1 cm and the frontal lobe was triangular in 80% of the cases. One patient had hypoplasia of corpus callosum.

CONCLUSIONS

MRI is helpful tool to demonstrate abnormalities of the olfactory system which are always present among patients suffering from Kallmann syndrome. MRI can also show, at the same time, a possible associated brain abnormality.

The dual positional maize lipoxygenase-1 was introduced into rice and T2 transgenic plants were produced. Cellular location of maize lipoxygenase-1 in transgenic rice and effects of calcium ion on membrane association in vitro were analyzed. Localization study by confocal microscopic analysis indicated that the maize lipoxygenase-1 was localized in cytoplasm. Sucrose-density fractionation experiment and in vitro protein transport to chloroplast showed that the maize lipoxygenase-1 can be associated with chloroplast. Secondary structure alignment revealed putative calcium binding sites in the PLAT domain of maize lipoxygenase-1 and the association of the maize lipoxygenase-1 with membranes was mediated by calcium ion in vitro. Our results provide evidences for calcium-mediated translocation of dual positional LOX without chloroplast targeting sequence from cytoplasm to chloroplast in plants for the first time.

OBJECTIVE

The novel biplanar X-ray unit "EOS", EOS Imaging, allows to acquire simultaneously 2 perpendicular planes of full-length spine and limbs and to measure spatially correct angles based on the acquired image data sets. This is to be seen alongside with a low spatial resolution, high investment costs and high operating expenses. The use of the biplanar X-ray unit in morphology based scelettal radiography might improve the cost-benefit-relation. Thus, the purpose of this study was to compare image quality of the EOS-unit and the flat panel (FP)-technology as reference in a clinical setting.

METHODS

All 114 patients of the Orthopedic Hospital Dept., who had a biplanar full-length lower limb radiograph and a FP-examination of the pelvis and/or the knee with maximum time interval of 3 months without changes in the clinical and radiological findings were included in the study. All X-ray examinations had been carried out due to clinical indications. Secondary captures comparable to the FP-images were extracted from the electronic EOS-image data sets. 4 radiologists independently from each other compared the visualization of normal anatomical structures of the pseudonymous EOS- and FP-images in a randomized order.

RESULTS

In the overwiew of all readers and all sceletal regions image quality of the FP-images was considered being superior in a mean of 83 ± 13 % standard deviation of the pair comparisons (minimum 48 %, maximum 100 %). Image quality of the EOS-images was assessed as being superior in 2 ± 3 % of the cases (0 %, 10 %). Image quality of 0.8 ± 3 % of the FP-images (0 %, 17 %) and 30 ± 34 % (0 %, 100 %) of the EOS-images was estimated as diagnostically inadequate. 30 ± 33 % of the pair comparisons (0 %, 100 %) showed a diagnostically inadequate image quality of the EOS-images and a diagnostically good image quality of the FP-images.

CONCLUSIONS

Image quality of biplanar full-length lower limb X-ray examinations is not suitable to be used for the diagnostic assessment of the morphological bone structure using the currently available technological setting.

CONCLUSIONS

▶ biplanar full-length lower limb X-ray examinations ▶ plat-panel radiography ▶ image quality.

A questionnaire and observational study was conducted to determine the problems associated with construction, design and hygiene methods used to maintain pit latrines in a high-density urban township in Malawi. The survey comprised 100 randomly selected households and the person responsible for the latrine maintenance was interviewed. Eighty-seven percent of householders used traditional latrines, 67% with earthen and 20% with concrete (i.e. sanitation platform, also known as San-plats) floors. A variety of makeshift squat-hole covers (e.g. iron sheets, rocks, cardboard) are used by those who cannot afford San-plats to reduce fly and odour problems. Most squat-hole covers were fouled with faecal matter and some presented risk of invasive injury due to sharp edges. Five households used ventilated improved latrines whilst the remaining eight per cent made use of their neighbour's latrine. Ownership of property, poverty and theft in the area significantly influenced the type and structural condition of the latrine. Walls, ceilings and doors of the superstructure comprised a mixture of miscellaneous materials (i.e. cardboard, paper, cloth, brick) making the hygienic upkeep of the latrine difficult. Cloth doors were often used for hand drying. Geographical location, such as rocky terrain with inadequate soil consolidation, and environmental conditions, such as heavy rains and floods, exacerbate structural defects. Coupled with the lack of attention afforded to the disposal of young children's faeces, wash-water and pit surcharging, the potential exists for widespread contamination of the surrounding environment and transmission of faecal-oral disease. More than half of the respondents stressed that they had not received any information regarding health education and hygienic maintenance of the latrine. As such, there is need for a hygiene education programme to promote awareness of appropriate sanitation and behavioural change. Cultural and religious factors were found not to influence latrine use.

This retrospective observational study evaluated cost effectiveness of first-line treatment of advanced nonsquamous non-small cell lung cancer (NSCLC) with pemetrexed/platinum (Pem/Plat) relative to paclitaxel/carboplatin (Pac/Carbo) and paclitaxel/carboplatin/bevacizumab (Pac/Carbo/Bev). Patients initiating first-line treatment from 2006 to 2009 were identified in electronic medical records of 20 US oncology practices. Pem/Plat patients were matched 1:1 on important characteristics with Pac/Carbo and Pac/Carbo/Bev patients and followed for 1 year to assess progression, survival, and costs. Bootstrapping was used to calculate the probability of falling within quadrants of the incremental cost-effectiveness plane. Kaplan-Meier analysis and Cox proportional hazards regression modeling were also performed. Three hundred Pem/Plat patients (mean age, 67.6 years; male, 56.0%; PS 0/1, 71.0%) were matched with 300 patients in the other cohorts. Median PFS was 134 days (Pem/Plat) versus 106 days (Pac/Carbo) (hazard ratio [HR]: 0.67, P < 0.001) and 126 days (Pac/Carbo/Bev) (HR: 0.68, P < 0.001). Median OS was 298 days (Pem/Plat) versus 218 days (Pac/Carbo) (HR: 0.88, P = 0.08) and 271 days (Pac/Carbo/Bev) (HR: 0.93, P = 0.31). Pem/Plat therapy costs were higher versus Pac/Carbo ($21,841 higher PFS; $19,137 higher OS; P ≤ 0.05) and lower versus Pac/Carbo/Bev ($15,160 lower PFS; $19,946 lower OS; P ≤ 0.05). Pem/Plat had a greater probability of higher costs/higher effectiveness versus Pac/Carbo (PFS, 90.1%; OS, 96.3%) and lower costs/higher effectiveness versus Pac/Carbo/Bev (PFS, 69.5%; OS, 85.0%). Pem/Plat had higher cost and effectiveness than Pac/Carbo; depending on a payer's or society's willingness to pay, Pem/Plat may be considered cost effective compared with Pac/Carbo. Pem/Plat yielded greater effectiveness with lower costs than Pac/Carbo/Bev.

African Americans are 40% more likely to be afflicted with hypertension than are non-Hispanic, white Americans, resulting in a 30% higher instance of mortality due to cardiovascular disease. There is debate about the relative contributions of genetic and sociocultural risk factors to the racial disparity in hypertension. We assayed three Alu insertion polymorphisms located in the ACE (angiotensin 1 converting enzyme), PLAT (plasminogen activator, tissue), and WNK1 (lysine deficient protein kinase 1) genes. We also estimated West African genetic ancestry and developed novel measures of perceived discrimination to create a biocultural model of blood pressure among African American adults in Tallahassee, Florida (n = 158). When tested separately, the ACE Alu noninsertion allele was significantly associated with higher systolic and diastolic blood pressure. In multiple regression analyses, West African genetic ancestry was not associated with blood pressure and reduced the strength of all blood pressure models tested. A gene × environment interaction was identified between the ACE Alu genotype and a new measure of unfair treatment that includes experiences by individuals close to the study participant. Inclusion of the WNK1 Alu genotype further improved this model of blood pressure variation. Our results suggest an association of the ACE and WNK1 genotypes with blood pressure that is consistent with their proposed gene functions. Measures of perceived unfair treatment of others show a threshold effect, with increased blood pressure occurring at higher values. The interaction between the ACE genotype and unfair treatment highlights the benefits of including both genetic and cultural data to investigate complex disease.

Exercise HR recovery (HRR) has proven an effective clinical means to assess parasympathetic dysfunction linked to all-cause mortality, but an analogous functional assessment for sympathetic dysfunction has not been developed.

OBJECTIVE

We investigated whether exercise recovery provides additional cardiorespiratory information, beyond the initial HRR period, to index sympathetic overactivity associated with insulin resistance.

METHODS

Young people (N = 20) with diverse percent body fat (9%-52%) were studied using fasting, oral glucose tolerance test (OGTT), and high-carbohydrate meal measurements. Participants also completed a graded fitness test (oxygen consumption peak test on cycle ergometer) after which HR and oxygen consumption (V x O2) measurements were continued for 3 min into recovery. The first, rapid phase of exercise recovery was used as the clinical measurement for parasympathetic control (HRR = HR2 min - HRmax). The second, initial plateau phase of exercise recovery was used to calculate a novel functional index for sympathetic overactivity (the plateau value for the ratio of HR normalized for V x O2 (HR/V x O2 plat)).

RESULTS

As expected, parasympathetic function (HRR) was within the normal range in these young people (-58 +/- 2 bpm). The index for sympathetic overactivity varied over a wide range from 9 to 34 bpm/(mL x kg x min(-1)), with obese adolescents having values in the highest 25th percentile. We found that this simple index was correlated to both the OGTT-derived whole-body insulin sensitivity index (r = -0.74, P < 0.001) and Homeostasis Assessment Model for Insulin Resistance (r = 0.76, P < 0.001), independent of percent body fat and parasympathetic function. Meal-induced thermogenesis was also associated with HR/V x O2 plat (r = -0.64, P < 0.01) but not with HRR.

CONCLUSIONS

In young individuals, recovery from intense exercise may provide a simple means to quantify both parasympathetic and sympathetic function. The exercise recovery index for sympathetic overactivity was linked to insulin resistance.

Repeated binge-like alcohol intoxication (RBAI) induces whole-body insulin resistance, which is predicted to increase the risk for metabolic syndrome and type 2 diabetes. Previously, we showed that acute alcohol intoxication increases mesenteric lymphatic permeability, perilymphatic adipose tissue (PLAT) inflammation, and circulating lipopolysaccharide levels in rats. We hypothesize that mesenteric lymphatic hyperpermeability, adipose tissue inflammation and associated dysregulated adipokine expression, and insulin signaling are central mechanisms underlying whole-body metabolic dysregulation resulting from RBAI. To test this hypothesis, male Sprague-Dawley rats surgically fitted with an intragastric catheter received a bolus of 2.5 g/kg/day of alcohol (12.5% alcohol w/v) or isocaloric dextrose in Vanilla Ensure (116 kcal/kg/day) for 3 days. Mesenteric lymphatic permeability, mesenteric (MFAT = PLAT) and subcutaneous (SFAT) adipose tissue inflammatory milieu, circulating adipokines, and markers of insulin responsiveness (pAKT and PTP1B protein expression) were determined following the last alcohol/dextrose administration. RBAI resulted in increased lymphatic permeability, MFAT-specific expression of inflammatory cytokines and markers of inflammatory cells (macrophages, dendritic, and T cells), decreased circulating adiponectin and visfatin levels, and MFAT-specific attenuation of insulin-stimulated protein kinase B phosphorylation (Ser) compared with dextrose-treated control animals. These results suggest that RBAI-induced mesenteric lymphatic hyperpermeability promotes inflammatory milieu, decreased insulin-sensitizing adipokines, and impaired insulin signaling in MFAT, which we propose may be an early event preceding systemic metabolic dysregulation. We speculate that RBAI-induced increase in gut-derived toxins, promoting lymphatic leak, and MFAT inflammatory milieu are mechanisms deserving further investigation to elucidate lymphatic-MFAT crosstalk events that precede and predispose for alcohol-induced insulin resistance.

Recent studies have demonstrated that carbon nanotubes (CNTs) induce platelet aggregation, endothelial dysfunction and vascular thrombosis. However, there is little information on the effects of CNTs on fibrinolysis. We investigated the role of pristine-commercial single-walled carbon nanotubes (SWCNTs) with <3% Co content in fibrinolysis and their contribution to the induction of pro-thrombotic processes in human vein endothelial cells (HUVEC). SWCNTs alone produced concentration-dependent oxidation, as measured by a dithiothreitol oxidation assay. Internalized SWCNTs were located in HUVEC treated with 25 μg/ml using transmission electron microscopy, whereas treatment with 50 μg/ml compromised cell viability, and oxidative stress increased significantly at 5 μg/ml. The study showed that in HUVEC treated with 25 μg SWCNT/ml, fibrinolysis-related gene expression and protein levels had increased by 3-12 h after treatment (serpine-1: 13-fold; PLAT: 11-fold and PLAU: 2-fold), but only the PAI-1 protein was increased (1.5-fold), whereas tissue and urokinase plasminogen activator proteins (tPA and uPA, respectively) tended to decrease. In summary, pristine SWCNTs treatment resulted in evident HUVEC damage caused by cell fiber contact, internalization, and oxidative stress due to contaminant metals. The generation of endothelial dysfunction, as shown by the altered expression of genes and proteins involved in fibrinolysis, suggest that SWCNTs display pro-thrombotic effects.

OBJECTIVE

We aimed to investigate the complications, surgical site infection (SSI), and survival in salvage surgery without free-flap reconstruction for patients with head and neck squamous cell carcinoma who were treated by platinum-based chemoradiotherapy (Plat-CRT) or cetuximab-based bioradiotherapy (Cet-BRT).

METHODS

Thirty-three patients treated by Plat-CRT and six treated by Cet-BRT had salvage surgery. We categorized postoperative complications according to the Clavien-Dindo classification and SSI according to the wound grading scale. Overall survival calculated by Kaplan-Meier method.

RESULTS

Patients with Cet-BRT were significantly associated with the presence of SSI (P<0.01) and grades IIIb-V of the Clavien-Dindo classification (P<0.01) compared with those with Plat-CRT. Patients with Cet-BRT had a significantly lower overall survival than those with Plat-CRT (P<0.05).

CONCLUSIONS

We demonstrated that patients with Cet-BRT were significantly more associated with the presence of SSI and grades IIIb-V in the Clavien-Dindo classification than those with CRT.