Enhanced sensitivity for peptide mapping with electrospray liquid chromatography-mass spectrometry in the presence of signal suppression due to trifluoroacetic acid-containing mobile phases.
A method is described for improving the sensitivity of peptide mapping with electrospray liquid chromatography--mass spectrometry using trifluoroacetic acid (TFA) containing HPLC mobile phases. The signal suppressing effects of TFA are shown to be due to the combined effect of ion-pairing and surface tension modifications. The post-column addition of a propionic acid-2-propanol (75:25, v/v) in a 1:2 proportion with the HPLC mobile phase counteracts the deleterious effects of TFA resulting in 10-100 x improvement of the signal-to-noise ratio. The system described introduces total HPLC flow (plus additive) directly into the electrospray source without splitting. Using 2.1 mm I.D. HPLC columns, minimum detectable quantities are below 40 pmol total protein. As examples, separations of proteolytic enzyme digests of several proteins are shown using standard HPLC conditions, comparing results with and without the addition of propionic acid. The application of the technique is shown in more depth in the identification of oxidative modification sites in glutamine synthetase. In this application, the enhanced sensitivity allowed location of a modified residue by comparison endoproteinase Lys C digest of native and oxidized forms of the protein without extensive sample preparation or concentration. A third application demonstrates the identification of glycosylation sites in an endoproteinase Arg C digest of single-chain plasminogen activator through the use of in-source collisionally induced dissociation.


Long-term kidney function in children with Wilms tumour and constitutional WT1 pathogenic variant.
Wilms tumour (WT) survivors, especially patients with associated syndromes or genitourinary anomalies due to constitutional WT1 pathogenic variant, have increased risk of kidney failure. We describe the long-term kidney function in children with WT and WT1 pathogenic variant to inform the surgical strategy and oncological management of such complex children. Retrospective analysis of patients with WT and constitutional WT1 pathogenic variant treated at a single centre between 1993 and 2016, reviewing genotype, phenotype, tumour histology, laterality, treatment, patient survival, and kidney outcome. We identified 25 patients (60% male, median age at diagnosis 14 months, range 4-74 months) with WT1 deletion (4), missense (2), nonsense (8), frameshift (7), or splice site (4) pathogenic variant. Thirteen (52%) had bilateral disease, 3 (12%) had WT-aniridia, 1 had incomplete Denys-Drash syndrome, 11 (44%) had genitourinary malformation, and 10 (40%) had no phenotypic anomalies. Patient survival was 100% and 3 patients were in remission after relapse at median follow-up of 9 years. Seven patients (28%) commenced chronic dialysis of which 3 were after bilateral nephrectomies. The overall kidney survival for this cohort as mean time to start of dialysis was 13.38 years (95% CI: 10.3-16.4), where 7 patients experienced kidney failure at a median of 5.6 years. All of these 7 patients were subsequently transplanted. In addition, 2 patients have stage III and stage IV chronic kidney disease and 12 patients have albuminuria and/or treatment with ACE inhibitors. Four patients (3 frameshift; 1 WT1 deletion) had normal blood pressure and kidney function without proteinuria at follow-up from 1.5 to 12 years. Despite the known high risk of kidney disease in patients with WT and constitutional WT1 pathogenic variant, nearly two-thirds of patients had sustained native kidney function, suggesting that nephron-sparing surgery (NSS) should be attempted when possible without compromising oncological risk. Larger international studies are needed for accurate assessment of WT1genotype-kidney function phenotype correlation.


Functional resistance activities to impact frailty: A protocol for a randomized controlled trial involving home care aide and frail older adult dyads.
A growing number of older adults use in-home Medicaid Waiver Home and Community Based services (HCBS) to facilitate aging-in-place. A primary service of this program is Home Care Aide assistance with activities of daily living and homemaker needs. Despite the known benefits of exercise, exercise programs are currently not offered to clients in the Medicaid Waiver system. Thus, the purpose of this paper is to describe a six-month Home Care Aide-led resistance exercise intervention protocol for frail older adults receiving Medicaid waiver services. A randomized controlled trial will be used. We will enroll 126 Home Care Aide-client dyads for a 6-month exercise intervention. The intervention will consist of training phases to promote muscle strength, power, and endurance. We will use an intention to treat principle using mixed effects models for the quantitative outcomes. To analyze qualitative outcomes, we will use conventional content analysis to examine themes from participant program evaluations. As greater numbers of adults age in place with frailty and employ Home Care Aides to help manage functional limitations, interventions embedded within usual care services play a critical role in bringing exercise into the home setting. The research described in this protocol will provide important knowledge about the impact of a Home Care Aide-led exercise intervention in reducing frailty in older adults. ClinicalTrials.gov Identifier: NCT02942992.


Induction chemotherapy followed by concurrent chemoradiotherapy vs. concurrent chemoradiotherapy for locoregionally advanced nasopharyngeal carcinoma: a retrospective cohort study.
To evaluate the efficacy and toxicity of induction chemotherapy followed by concurrent chemoradiotherapy vs. concurrent chemoradiotherapy for locoregionally advanced nasopharyngeal carcinoma (NPC). We reviewed data of locoregionally advanced NPC patients who underwent 2 different treatment plans, 1 with induction chemotherapy followed by concurrent chemoradiotherapy (IC + CCRT) and the other with only concurrent chemoradiotherapy (CCRT). All patients received cisplatin 80 mg/m2 3 weeks one cycle concurrently with intensity-modulated radiation therapy, and three IC protocols were included for the IC + CCRT group. Data of 262 patients treated from May 2011 to November 2014 were found eligible for our study. With a median follow-up of 29.02 months, no significant differences were detected between the two groups on the 2-year overall survival or OS rates (96.63 vs. 92.86%, P = 0.169), 2-year distant metastasis-free survival or DMFS rates (91.57 vs. 86.90%, P = 0.246) and 3-year DMFS rates (90.45 vs. 82.14%, P = 0.093). However, they were statistically different on 2-year locoregional failure-free survival or LFFS rates (94.94 vs. 86.90%, P = 0.020), 3-year OS rates (95.51 vs. 82.14%, P = 0.002) and 3-year LFFS rates (93.82 vs. 79.76%, P = 0.001). The most common severe (grade 3-4) adverse events we recorded were Nausea/vomiting (36.52 vs. 33.33%), leukopenia (14.04 vs. 4.76%), thrombocytopenia (3.93 vs. 3.57%) and hepatic toxicity (3.37 vs. 2.38%). Compared with CCRT, the treatment plan IC + CCRT produced significantly encouraging outcomes in locoregionally advanced NPC patients on local progression-free survival and 3-year overall survival position, but might raise the risk of certain adverse reactions.


High larvicidal efficacy of yeast-encapsulated orange oil against Aedes aegypti strains from Brazil.
Botanical substances such as essential oils (EOs) have demonstrated insecticidal properties and are a valid option for vector control. However, free EOs are unreliable as mosquito larvicides due their easy degradation by environmental exposure to ultraviolet light and higher temperatures. Here, we assessed the efficacy of a mosquito larvicide based on orange oil in a yeast-based delivery system against Aedes aegypti strains with different resistance status towards chemical neurotoxic insecticides. This larvicide preparation was physicochemically characterized in a previous report. Larvae of four Ae. aegypti strains from different regions of Brazil and different resistance profiles for deltamethrin (pyrethroid) and temephos (organophosphate) were tested against yeast-encapsulated orange oil (YEOO) in laboratory conditions for measurement of LC50 and LC90 values. The same assays were performed with the Belo Horizonte strain under environmental conditions (natural light and temperature). The resistance profiles of these strains were compared to the Rockefeller reference strain in all conditions. YEOO was found to be a highly active larvicide (LC50 < 50 mg/L) against all Ae. aegypti strains tested in both laboratory conditions (LC50 = 8.1-24.7 mg/L) and environmental conditions with natural light and temperature fluctuation (LC50 = 20.0-49.9 mg/L). Moreover, all strains were considered susceptible (RR < 5) to YEOO, considering resistance ratios calculated based on the Rockefeller strain. The resistance ratios were only higher than 2.5 for LC90-95 of Belo Horizonte in the laboratory, probably due the higher heterogeneity associated with older egg papers (> 5 months). YEOO demonstrates high larvicidal activity against Ae. aegypti strains with resistant phenotypes for deltamethrin (PY) and temephos (OP). This larvicidal activity suggests the potential for the development of YEOO as an alternative intervention to synthetic insecticides in integrated vector management programs, for populations with resistance to commonly used insecticides.


Visible-Light-Active Unsymmetrical Squaraine Dyes with Pyridyl Anchoring Groups for Dye-Sensitized Solar Cells.
Visible-light-active alkyl group-wrapped unsymmetrical squaraine dyes SD1-SD3 were synthesized, featuring an indoline donor and pyridine and carboxylic acid anchoring groups. Their photophysical, electrochemical, and photovoltaic characteristics were examined by fabricating a dye-sensitized solar cell (DSSC) device. Both carboxylic acid and pyridine anchoring groups containing squaraine dyes SD3 and SD2 possess similar photophysical and electrochemical characteristics. However, their photovoltaic performances were completely different. The SD3 dye with the carboxylic acid anchoring group displayed a DSSC device efficiency of 7.20% (VOC 0.81 V; JSC 12.29 mA/cm2) using iodolyte (I-/I3-) electrolyte, compared to SD1 (VOC 0.659 V; JSC 4.97 mA/cm2; and η - 2.34%) and SD2 (VOC 0.629 V; JSC 1.68 mA/cm2; and η - 0.84%), which were featured with pyridyl anchoring groups. These results were attributed to dye loading on the Lewis and Brønsted acidic sites of TiO2 and the importance of aggregated structures for photocurrent generation. In the incident photon-to-current efficiency (IPCE) analysis, SD1 dye-sensitized devices exhibited photocurrent generation from both monomeric and aggregated dyes on the TiO2 surface. In contrast, SD2 showed photocurrent generation solely from aggregated states. Despite the introduction of long alkyl chains to reduce dye aggregation and charge recombination, the results indicated preferential charge injection from only the aggregated SD2 dye on TiO2. Fluorescence-quenching experiments indicated an efficient charge transfer from the aggregated SD2 dye to TiO2 compared to that of the monomeric dye. Cosensitization, a method to enhance the light-harvesting efficiency and photocurrent generation in DSSCs, was explored by simultaneously cosensitizing pyridyl-based dyes (SD1 and SD2) with a blue-colored carboxylic acid-based squaraine dye SD4. IPCE analysis demonstrated that both SD1 and SD4 contributed to generating a photocurrent of 9.11 mA/cm2. The sequential cosensitization of SD1 and SD4 with the coadsorbent CDCA showed the highest performance, with a VOC of 0.663 V, a JSC of 11.43 mA/cm2, and an efficiency (η) of 5.20%.


A comparison of outcomes after lung transplantation between European and North American centers.
With advancements in basic science and clinical medicine, lung transplantation (LT) has evolved rapidly over the last three decades. However, it is unclear if significant regional variations exist in long-term outcomes after LT. To investigate potential differences, we performed a retrospective, comparative cohort analysis of adult patients undergoing deceased donor single or double LT in North America (NA) or Europe between January 2006 and December 2016. Data up to April 2019 were abstracted from the International Society for Heart and Lung Transplantation (ISHLT) Thoracic Organ Registry. We compared overall survival (OS) between North American and European LT centers in a propensity score matched analysis. In 3,115 well-matched pairs, though 30-day survival was similar between groups (NA 96.2% vs Europe 95.4%, p = 0.116), 5-year survival was significantly higher in European patients (NA 60.1% vs Europe 70.3%, p < 0.001). This survival difference persisted in a sensitivity analysis excluding Canadian patients. Prior observations suggest that these disparities are at least partly related to better access to care via universal healthcare models prevalent in Europe. Future studies are warranted to confirm our findings and explore other causal mechanisms. It is likely that potential solutions will require concerted efforts from healthcare providers and policymakers.


Regulation of kallikrein and renin release by the isolated perfused rat kidney.
Rat kidneys perfused in vitro released kallikrein in urine, and renin and kallikrein in the perfusate. The kallikrein was characterized by its kininogenase activity and released bradykinin from bovine and dog substrates. Inactive trypsin activatable kallikrein was present in both perfusate and urine. Kallikrein secretion in urine was influenced by changes in perfusion pressure (PP). Raising the PP strikingly increased urinary kallikrein and lowering PP reduced it. Urinary water and electrolyte output were augmented to the same extent by furosemide and mannitol administration as by raising the PP, but neither drug affected kallikrein. Isoproterenol stimulated the release of renin but not kallikrein. Stopping the oxygen supply to the perfusate suppressed kallikrein secretion in urine and renin release in the perfusate. The kidneys released ten times less kallikrein in the perfusate than in urine, and perfusate kallikrein was not influenced by changes in PP. It is concluded that in this model, changes in PP and/or renal blood flow and/or oxygen supply regulate kallikrein secretion in urine, but that this secretion is unaffected by changes in urinary output. We also conclude that kallikrein release in urine and renin release in perfusate are regulated simultaneously by renal hemodynamic changes but are not affected concomitant by beta-adrenergic stimulation or changes in distal urine composition.


Assessing Impact of Platinum Complexes on Mitochondrial Functions.
Platinum-based antitumor drugs play important roles in the clinical treatment of various tumors. Nevertheless, some deficiencies such as poor targeting ability, low bioavailability, in vivo deactivation, drug resistance, and side effects undermine the efficacy of these drugs. Mitochondria are important organelles which regulate the energy metabolism, physiological function, life span, and survival of the cells. Regulating or interfering with mitochondrial metabolism is of great significance in the prevention or treatment of cancers. Thus, a series of mitochondrion-targeted platinum complexes were prepared by modifying triphenylphosphine (TPP+) through chemical modifications, which endow traditional platinum drugs with new properties and mechanisms through interfering with mitochondrial DNA (mtDNA), mitochondrial membrane potential (MMP), mitochondrial morphology, mitochondrial bioenergetics, or production of reactive oxygen species (ROS), thereby opening a new path for the clinical application of platinum drugs. Here we introduce the synthesis of some TPP+-modified platinum (II, IV) complexes in details and the detection method of the activity parameters related to the mitochondrial functions.


Disposition kinetics and distribution of demeclocycline in various biological fluids in female goats.
A pharmacokinetic study of demeclocycline was carried out following intravenous administration at 5 mg/kg body weight in lactating goats. Demeclocycline appeared within 5 min in plasma, interstitial fluid (isf) and urine, while it appeared at 1 h in milk. Peak concentrations of 21.70 +/- 4.06, 2.67 +/- 0.23, 5.65 +/- 0.45 and 82.23 +/- 10.06 micrograms/ml were attained at 5 min and at 6, 8 and 8 h in plasma, isf, milk and urine respectively. A potentially therapeutic concentration of greater than or equal to 0.5 micrograms/ml was maintained from 5 min-36 h, 30 min-30 h, 1-36 h and 5 min-48 h in plasma, isf, milk and urine respectively. The drug was detectable in all the above biological fluids for at least 48 h. A low distribution half life (t1/2 alpha) of 0.44 +/- 0.04 h and a high elimination half life (t1/2 beta) of 19.24 +/- 1.22 h denote rapid distribution but very slow elimination of the drug in goats. A high tissue plasma concentration ratio [K12:(K21-beta)] of 5.12 +/- 0.97 during the elimination phase and a Vdarea of 1.59 +/- 0.18 L/kg indicate uniform distribution of demeclocycline in the tissues and body fluids of goats. The dosage regimen for maintaining minimum plasma concentration (Cp infinity min = MIC) of 0.5, 1.0 and 1.5 micrograms/ml at selected dosage intervals of 12 and 24 h was also calculated.


In Situ Thermal Cross-Linking of 9,9'-Spirobifluorene-Based Hole-Transporting Layer for Perovskite Solar Cells.
A novel 9,9'-spirobifluorene derivative bearing thermally cross-linkable vinyl groups (V1382) was developed as a hole-transporting material for perovskite solar cells (PSCs). After thermal cross-linking, a smooth and solvent-resistant three-dimensional (3D) polymeric network is formed such that orthogonal solvents are no longer needed to process subsequent layers. Copolymerizing V1382 with 4,4'-thiobisbenzenethiol (dithiol) lowers the cross-linking temperature to 103 °C via the facile thiol-ene "click" reaction. The effectiveness of the cross-linked V1382/dithiol was demonstrated both as a hole-transporting material in p-i-n and as an interlayer between the perovskite and the hole-transporting layer in n-i-p PSC devices. Both devices exhibit better power conversion efficiencies and operational stability than devices using conventional PTAA or Spiro-OMeTAD hole-transporting materials.


Great expectations: minor differences in initial instructions have a major impact on visual search in the absence of feedback.
Professions such as radiology and aviation security screening that rely on visual search-the act of looking for targets among distractors-often cannot provide operators immediate feedback, which can create situations where performance may be largely driven by the searchers' own expectations. For example, if searchers do not expect relatively hard-to-spot targets to be present in a given search, they may find easy-to-spot targets but systematically quit searching before finding more difficult ones. Without feedback, searchers can create self-fulfilling prophecies where they incorrectly reinforce initial biases (e.g., first assuming and then, perhaps wrongly, concluding hard-to-spot targets are rare). In the current study, two groups of searchers completed an identical visual search task but with just a single difference in their initial task instructions before the experiment started; those in the "high-expectation" condition were told that each trial could have one or two targets present (i.e., correctly implying no target-absent trials) and those in the "low-expectation" condition were told that each trial would have up to two targets (i.e., incorrectly implying there could be target-absent trials). Compared to the high-expectation group, the low-expectation group had a lower hit rate, lower false alarm rate and quit trials more quickly, consistent with a lower quitting threshold (i.e., performing less exhaustive searches) and a potentially higher target-present decision criterion. The expectation effect was present from the start and remained across the experiment-despite exposure to the same true distribution of targets, the groups' performances remained divergent, primarily driven by the different subjective experiences caused by each groups' self-fulfilling prophecies. The effects were limited to the single-targets trials, which provides insights into the mechanisms affected by the initial expectations set by the instructions. In sum, initial expectations can have dramatic influences-searchers who do not expect to find a target, are less likely to find a target as they are more likely to quit searching earlier.


The circadian clock controls sunburn apoptosis and erythema in mouse skin.
Epidemiological studies of humans and experimental studies with mouse models suggest that sunburn resulting from exposure to excessive UV light and damage to DNA confers an increased risk for melanoma and non-melanoma skin cancer. Previous reports have shown that both nucleotide excision repair, which is the sole pathway in humans for removing UV photoproducts, and DNA replication are regulated by the circadian clock in mouse skin. Furthermore, the timing of UV exposure during the circadian cycle has been shown to affect skin carcinogenesis in mice. Because sunburn and skin cancer are causally related, we investigated UV-induced sunburn apoptosis and erythema in mouse skin as a function of circadian time. Interestingly, we observed that sunburn apoptosis, inflammatory cytokine induction, and erythema were maximal following an acute early-morning exposure to UV and minimal following an afternoon exposure. Early-morning exposure to UV also produced maximal activation of ataxia telangiectasia mutated and Rad3-related (Atr)-mediated DNA damage checkpoint signaling, including activation of the tumor suppressor p53, which is known to control the process of sunburn apoptosis. These data provide early evidence that the circadian clock has an important role in the erythemal response in UV-irradiated skin. The early morning is when DNA repair is at a minimum, and thus the acute responses likely are associated with unrepaired DNA damage. The prior report that mice are more susceptible to skin cancer induction following chronic irradiation in the AM, when p53 levels are maximally induced, is discussed in terms of the mutational inactivation of p53 during chronic irradiation.


Laparoscopic lymph node dissection after endoscopic submucosal dissection: a novel and minimally invasive approach to treating early-stage gastric cancer.
Endoscopic submucosal dissection (ESD), a newly developed endoscopic mucosal resection (EMR) technique, can completely cure a differentiated mucosal gastric cancer smaller than 2 cm. For early-stage gastric cancers (EGCs) deviating from the above-mentioned criterion, gastrectomy with lymph node dissection is performed for potential risk of lymph node metastasis (LNM). However, many of surgical EGC cases actually do not have LNM, indicating this surgery may not be necessary for many cases of EGC. To avoid this unnecessary surgery, we have introduced laparoscopic lymph node dissection (LLND) after ESD. Standard gastrectomy with extended lymph node dissection is indicated for patients if LLND reveals LNM. We present our novel approach and the preliminary results of EGC patients having potential risk of LNM. Five patients with EGC deviating from the EMR criterion underwent the combination of ESD and LLND. ESD was performed using a newly developed insulation-tipped diathermic knife. Lymph nodes, which were determined on the basis of the location of the primary tumor and lymphatic drainage of the stomach, were removed laparoscopically. The lymphatic drainage was visualized by submucosally injecting indocyanine green (ICG) around the post-ESD ulcerative scars during intraoperative gastroscopy. The ESD enabled en bloc resection without any complications. The resected margins of all the lesions were free of cancer cells vertically and horizontally. LLND was successfully performed without any complications. The mean number of the dissected lymph nodes was 15 (range 6 to 22). In 4 of the 5 patients, the dissected lymph nodes were free of cancer cells, and therefore, the combination of ESD and LLND was considered a definitive treatment. The remaining patient was found to have LNM but chose not to undergo any surgery. During follow-ups, the patients' previous quality of life was restored without any tumor recurrence. The combination of ESD and LLND enables the complete resection of the primary tumor and the histologic determination of lymph node status. This combination treatment is a potential, minimally invasive method, and may obviate unnecessary gastrectomy without compromising curability for EGC patients having the potential risk of LNM.


Stentless Interventional Procedure Using Rotational Atherectomy and Drug-Coated Balloon for Noncalcified De Novo Lesions.
Several recent reports have shown that a stentless interventional procedure using rotational atherectomy followed by drug-coated balloon (DCB) treatment (RA/DCB) is a potent revascularization therapy for calcified de novo lesions even in the new-generation drug-eluting stent era; however, the role of the RA/DCB procedure for noncalcified de novo lesions remains unclear. A total of 47 consecutive patients (53 lesions) who underwent RA/DCB for coronary de novo lesions were enrolled. According to the presence or absence of severe calcification at target lesions on fluoroscopy, the 47 patients were divided into the noncalcified cases (n = 12) and the calcified cases (n = 35), and the 53 lesions were divided into the noncalcified lesions (n = 14) and the calcified lesions (n = 39). The noncalcified cases tended to have a higher frequency of bleeding risk and had a significantly lower prevalence of dual antiplatelet therapy compared with the calcified cases. The main lesion-specific factors for the RA/DCB procedure among the noncalcified lesions were presence of left circumflex coronary artery ostial lesion. The final burr size, DCB diameter used, and angiographic success rate did not significantly differ between the 2 groups. The noncalcified lesions had a larger reference diameter and a shorter lesion length than the calcified lesions, whereas acute gain and late lumen loss did not differ between the 2 groups. Nine-month clinical outcomes were comparable between the 2 groups. Under drug-eluting stent-unsuitable clinical or lesion conditions, acute and midterm outcomes of RA/DCB for noncalcified de novo lesions might be comparable with those for calcified de novo lesions.


Transcription factor c-Maf deletion improves streptozotocin-induced diabetic nephropathy by directly regulating Sglt2 and Glut2.
The transcription factor c-Maf has been widely studied and has been reported to play a critical role in embryonic kidney development; however, the postnatal functions of c-Maf in adult kidneys remain unknown as c-Maf-null C57BL/6J mice exhibit embryonic lethality. In this study, we investigated the role of c-Maf in adult mouse kidneys by comparing the phenotypes of tamoxifen-inducible (TAM-inducible) c-Maf-knockout mice (c-Maffl/fl; CAG-Cre-ERTM mice named "c-MafΔTAM") with those of c-Maffl/fl control mice, 10 days after TAM injection [TAM(10d)]. In addition, we examined the effects of c-Maf deletion on diabetic conditions by injecting the mice with streptozotocin, 4 weeks before TAM injection. c-MafΔTAM mice displayed primary glycosuria caused by sodium-glucose cotransporter 2 (Sglt2) and glucose transporter 2 (Glut2) downregulation in the kidneys without diabetes, as well as morphological changes and life-threatening injuries in the kidneys on TAM(10d). Under diabetic conditions, c-Maf deletion promoted recovery from hyperglycemia and suppressed albuminuria and diabetic nephropathy by causing similar effects as did Sglt2 knockout and SGLT2 inhibitors. In addition to demonstrating the potentially unique gene regulation of c-Maf, these findings highlight the renoprotective effects of c-Maf deficiency under diabetic conditions and suggest that c-Maf could be a novel therapeutic target gene for treating diabetic nephropathy.


miRNAomics analysis reveals the promoting effects of cigarette smoke extract-treated Beas-2B-derived exosomes on macrophage polarization.
Inhalation of cigarette smoke induces airway and parenchyma inflammation that predisposes smokers to multiple lung diseases such as COPD. Macrophage polarization, an important specifying feature of inflammation, is involved in the progression of pulmonary inflammation. Exosomes and their loaded miRNAs provide a medium for cross-talk between alveolar macrophages and lung epithelial cells to maintain lung homeostasis. In this study, we treated Beas-2B with CSE to speculate the effects of Beas-2B-derived exosomes on macrophage polarization and performed exosomal miRNAomics analysis to explore the mechanism. We found that CSE-treated Beas-2B-derived exosomes could not only increase the percentages of CD86+, CD80+ CD163+, and CD206+ cells but also induce the secretion of TNF-α, IL-6, iNOS, IL-10, Arg-1, and TGF-β, indicating both M1 and M2 polarization of RAW264.7 macrophages were promoting. We performed miRNAomics analysis to identify 27 differentially expressed exosomal miRNAs such as miR-29a-3p and miR-1307-5p. Next, we obtained 14942 target genes of these miRNAs such as SCN1A and PLEKHA1 through the prediction of TargetScan and miRanda. We utilized KEGG enrichment analysis for these targets to identify potential pathways such as the PI3K-Akt signaling pathway and the MAPK signaling pathway on the regulation of macrophage polarization. We further found that miR-21-3p or miR-27b-3p may play critical roles in the promotion of CSE-Exo on macrophage polarization by miRNA interference. Collectively, this study provided novel information for diagnostic and therapeutic tactics of cigarette smoke-related lung diseases.


Multi-analyte calibration and verification of a multi-parameter laser-based pulse oximeter.
Almost since its introduction pulse oximetry was known to overestimate oxygen saturation in cases of carbon monoxide poisoning or elevated methemoglobin (metHb) levels. To eliminate this dangerous behavior some manufacturers have added additional LED emitters to try to increase the number of measured hemoglobin species and to improve measurement accuracy, but have not been very successful. We hypothesized that the use of narrow-band laser light sources would make accurate and precise measurement of the four primary species of hemoglobin possible, even in cases of elevated levels of carboxyhemoglobin (COHb). Calibration and verification studies were performed on a tissue simulator that employed an artificial finger pulsating with whole human blood. This simulator allowed safe generation of 165 different combinations of the levels of oxyhemoglobin (O2Hb), COHb, metHb, and reduced hemoglobin (RHb) for calibration of the laser-based pulse oximeter. A follow-on study used 56 mixed hemoglobin levels for verification and statistical analysis of the performance of this device. This laser-based pulse oximeter measured all four species of hemoglobin accurately and precisely (ARMS ≤ 1.8%) for metHb levels in the clinically normal range. At elevated metHb levels the device continued to provide accurate and precise measurements of metHb and RHb (ARMS ≤ 1.7%). The use of monochromatic laser light sources can create a new generation of highly accurate, multi-parameter, pulse oximeters.


Identification of two novel HLA-C alleles, Cw*070105 and Cw*1408, from East African women.
HLA-C was shown to be a highly polymorphic gene that can be accurately typed by sequencing methodologies. We report two novel HLA-C alleles identified during sequence-based typing of East African populations; the novel alleles were confirmed by sequencing two separate polymerase chain reaction products and by molecular cloning and sequencing multiple clones. The first new allele is identical to Cw*0701 except for a single-nucleotide synonymous substitution at codon 182 (GCA-->GCG). The new allele has been named by the WHO nomenclature committee as Cw*070105. The second new allele is identical to Cw*1403 except for a nonsynonomous change at codon 21 (CAC-->CGC), changed from histidine to arginine. The new allele has been named by the WHO nomenclature committee as Cw*1408.


Transpulmonary thermodilution cardiac output measurement using the axillary artery in critically ill patients.
To compare cardiac output (CO) as measured by the arterial thermodilution technique using only a central venous catheter and an arterial catheter inserted into the axillary artery, with conventional CO measurement with thermodilution using a pulmonary artery (PA) catheter (PAC). Prospective clinical study in which each patient served as his/her own control. General ICU of a large tertiary-care teaching hospital. 22 patients who required invasive hemodynamic monitoring in the ICU. CO measurements made using the PAC (COpa) were compared to bolus arterial thermodilution measurements (COax). The significance of acute changes in the continuous CO measurements during acute hemodynamic episodes was observed. The correlation between the two techniques (COpa and COax) was R(2) = 0.82. There was a tendency for 5% overestimation of COpa by the COax. The SEM% (SEM/average CO) for COax and COpa was 2.6% and 3.2%, respectively. The bias between measurements was 0.27 +/- 0.67 L/min, and the limits of agreement (mean difference +/- 2 SD) from minus 1.07 L/min to 1.63 L/min. In critically ill patients, in whom the measurement of CO is required, arterial thermodilution, using a central vein and the axillary artery is accurate and reproducible.


Patient Comorbidities Drive High Mortality Rates Associated with Major Liver Resections Irrespective of Hospital Volume.
Major hepatectomies are utilized to manage primary hepatic malignancies. Reports from high-volume centers (HVCs) with minimal perioperative mortality focus on multiple aspects of perioperative care, although patient-specific factors remain unelucidated. We identified patient factors associated with outcomes and examined whether these contribute to survival differences. We queried the National Cancer Database (2006-2015) for patients with primary liver malignancies managed with major hepatectomy. Facilities were dichotomized by volume (high volume: >15 hepatectomies/year). Perioperative outcomes were compared based on patient demographic and clinical characteristics as well as center volume. 4263 patients were included with 78.5% receiving care in low-volume centers (LVCs). 90-day postoperative mortality was higher in LVCs vs. HVCs (12% vs. 7.5%; P < .001). Factors associated with undergoing surgery in LVCs included: living in areas with lower income (P = .006) and education (P < .001), having nonprivate insurance (P < .001), residing near the care center (P < .001), and having a comorbidity score (CDS) >1 (P = .014). Patients with CDS ≤ 1 had higher 90-day mortality in LVCs (11.3% vs. 6.6%; P < .001) and had similar outcomes in LVCs and HVCs (15.6% vs. 13.7% P = .6). Patients with CDS > 1 were more likely to receive care in LVCs (16.3% vs. 12.7%; P < .001). Reduced perioperative mortality following major hepatectomy in HVCs is driven by optimal management of patients with low CDS. However, nearly 1 in 5 patients who undergo major hepatectomies have a high CDS and approximately 15% of them succumb in the perioperative period irrespective of the treating centers' experience.


[IOL choice predictability, 1-year visual outcome, and complications after cataract surgery in children].
To measure the refraction 1year after cataract surgery in children as a function of under-correction of the implant power and to evaluate the visual prognosis and possible therapeutic challenges. We conducted a retrospective study of 28 children (38 affected eyes) who underwent unilateral or bilateral cataract surgery with primary intraocular lens implantation over a one-year period of time. The age at the time of surgery was between 1month and 17years. Ten patients had bilateral cataracts (35%) and 18 patients unilateral (65%). Ten patients were between 0 and 2years of age, 10 patients between 2 and 6years of age, and 8 patients were over 6years old. 72 % of the cataracts were idiopathic. The cataracts were mainly cortico-nuclear (37%). Monocular acuity averaged 0.7 LogMAR (2/10) between 2 and 6years, and 0.2 LogMAR (6.3/10) after 6years. The percentage of under-correction was 31% between 0 and 2years, 12.2% between 2 and 6years, and 2.3% after 6years, for a mean under-correction of 15.2%. The power of the chosen implant after under-correction was 25 diopters on average. One year after surgery, visual acuity averaged 0.2 LogMAR (6.3/10) between 2 and 6years old,and 0.1 LogMAR (8/10) after 6years. The spherical equivalent remained globally stable after surgery (the mean postoperative value was+1.25 diopters). The highest rate of revision surgery occurred in the 0-2year-olds (40%). The most common complication was cellular proliferation (up to 80% of the 0-2year-olds). Only one case of ocular hypertension was noted. It is necessary to apply an under-correction of the intraocular lens power, decreasing with the age of the child. The gain in visual acuity is modest, and the main complication is cellular proliferation.


Reassignment of Western Australia Paracnephia gladiator Moulton Adler to a new genus, Bunyipellum (Diptera: Simuliidae).
With new material available of most stages of many known Australian Paracnephia, including new species, it is now clear that certain segregates warrant assignment to new genera. This applies to Paracnephia gladiator Moulton Adler, a Western Australia simuliid with numerous unique character states. The species is fully redescribed and assigned to Bunyipellum nov. gen. A diagnosis is provided and relationships discussed, as is historical biogeography. Bunyipellum appears to be more closely related to elements of the South American simuliid fauna than to any other Gondwanan Australian species.


Relationship of prior hormonal therapy to subsequent estramustine phosphate treatment in advanced prostatic cancer.
The relationship of prior hormonal therapy to subsequent response on estramustine phosphate (Estracyt) was examined in 107 patients with advanced prostatic cancer treated in two different Phase II chemotherapy trials. In both trials patients with the longest prior hormonal treatment were the least likely to respond to estramustine phosphate. Patients in the series from the National Prostatic Cancer Project with a response classification to prior hormonal therapy had only a 26 per cent response to subsequent estramustine phosphate therapy, whereas 40 per cent of those with no prior response to hormonal therapy responded to estramustine phosphate. This latter group had the shortest average disease duration from diagnosis. The sample of prostate cancers studied appeared to include groups that were sensitive to both hormones and cytotoxic activity as well as to either of these two alone. These data support the contention that estramustine phosphate may act both as an estrogenic and a cytotoxic agent.


Effect of lentinan on lymphocyte subsets of peripheral blood, lymph nodes, and tumor tissues in patients with gastric cancer.
The effect of lentinan on lymphocyte subsets of peripheral blood, lymph nodes, and tumor tissues in gastric cancer patients was investigated. A 2-mg dose of lentinan was administered to 12 patients intravenously twice, the first at 3-9 days before and the second the day before surgery. The results were then compared with a control group without lentinan administration comprising 12 patients. Regarding peripheral blood and lymph nodes without metastasis, lymphocyte subsets defined with anti-CD3, anti-CD4, anti-CD8, anti-Leu7, and anti-Leu11 were analyzed by flow-cytometry. As for tumor tissues, lymphocyte subsets defined with anti-CD3, anti-CD4, anti-CD8, anti-Leu11, and anti-M3 were analyzed after immunohistochemical staining. There were no significant changes in the lymphocyte subsets of peripheral blood between the two groups. In the lymph nodes, the CD4 cell ratios increased; otherwise in regard to tumor-infiltrating lymphocytes (TILs), the number of CD4, Leu11 and LeuM3 cells showed a prominent increase. Therefore, lentinan was observed to produce different effects in accordance with the subjective organs.


Kuwait's marine biodiversity: Qualitative assessment of indicator habitats and species.
The tropical waters of the Northern Arabian Gulf have a long history of maritime resource richness. High levels of biodiversity result from the complex matrix of coastal habitats, coral reefs and sea grass beds that characterise the region. Insight into the ongoing health of such habitats and the broader Kuwait maritime environment can be gauged by the status of indicator species found within these habitats. Here we review information on the occurrence, distribution and threats to key marine habitats and associated indicator species to provide an updated assessment of the state of the Kuwait's marine biodiversity. Critical evaluation of historic data highlights knowledge gaps needed inform the focus of future monitoring and conservation efforts. This assessment is designed to evaluate performance against environmental policy commitments, while providing a solid foundation for the design of comprehensive marine ecosystem management strategies.


Clinical and parasitological aspects of onchocercal skin diseases in Nigeria.
An assessment of onchocercal skin disease (OSD) conducted in 38 rural communities in the Imo River Basin, Nigeria, between March 1999 and September 2000, showed that depigmentation (DPM) was the most prevalent lesion in persons with skin microfilariae (mf) (26.3%), followed by chronic papular onchodermatitis (CPOD) (18.1%) and acute papular onchodermatitis (APOD) (15.5%). There was no significant difference (P > 0.05) in sex-related prevalence of OSD. While CPOD, lichenified onchodermatitis (LOD) and DPM were more prevalent in subjects over 30 years old, APOD was associated more with those aged less than 30 years. OSD occurred with concomitant itching in nearly 50% of subjects. The geometric mean intensity of infection was 13 mf/mg per skin snip.


Relationship between maternal nutritional status and infant's weight and body proportions at birth.
To examine maternal nutritional status and its relationship to infant weight and body proportions. Retrospective study of births from January-December 1990. University Hospital of the West Indies, Jamaica. Records for 2394 live, singleton births, between 200-305 d gestation. Birth weight, crown heel length, head circumference, ponderal index, head circumference:length ratio, placental weight, placental:birth weight ratio. Mothers who were lighter had babies who had lower birth weight, were shorter, had smaller heads and had a higher HC:L ratio. Shorter and thinner women had babies who had lower birth weights, were shorter, had smaller heads and lighter placentas. Thinner women also had babies with a lower placental:birth weight ratio, and their BMI's were not linearly related to ponderal index and HC:L ratio. Women whose first trimester Hb levels were < 9.5 g/dl had babies with the lowest birth weight, crown heel length, placental weight and ponderal index. These measurements increased as the Hb levels rose to 12.5 g/dl but then fell at Hb levels > 12.5 g/dl. In the second and third trimester Hb levels were negatively associated with birth weight, crown heel length, head circumference, placenta weight and ponderal index. The data support the hypothesis that poor maternal nutrition is associated with foetal growth restraint. Poor maternal nutrition as indicated by low weight, height, and BMI are associated with smaller, shorter babies with smaller heads. Haemoglobin levels > 12.5 g/dl in pregnancy are associated with lighter, shorter, thinner babies, with smaller heads.


Factors controlling the washout of the interstitial space of the isolated, perfused rat heart.
1. The time course of the washout of the extracellular markers, inulin, raffinose, sucrose and sorbitol, has been determined for the isolated rat heart perfused for 20 min with Krebs bicarbonate medium containing the marker, and washed out with marker-free perfusate.2. The filtration problem for the perfused rat heart was found to be due to omission of the gassing of the concentrated sodium bicarbonate solution with CO(2) in the preparation of the perfusate. This probably caused calcium phosphate to be precipitated. The reduced contractility of the heart was reflected in the washout of the extracellular markers.3. The logarithmic plot of the heart content of the marker against time showed two components. The slower was an exponential process, the rate of which was linearly related to the diffusion coefficient for the various markers.4. The faster component was absent in the quiescent heart, and in the beating heart its contribution to the washout of the interstitial space was related to the strength of contraction.5. Similar washout curves were obtained for Evans Blue-albumin conjugate, which readily penetrates the interstitial space of the isolated rat heart.6. It is suggested that the two components of efflux are a diffusion process limited by perfusate flow, and bulk movements of fluid.


Comparison of quartz and Teflon filters for simultaneous collection of size-separated ultrafine aerosol particles and gas-phase zero samples.
In this research, the two most common filter media, quartz and Teflon, were tested to obtain information about the possible adsorption of gas-phase compounds onto filters during long sample collection of atmospheric aerosols. Particles of nanometer-size for off-line chemical characterization were collected using a recently introduced differential mobility analyzer for size separation. Samples were collected at an urban site (Helsinki, SMEARIII station) during spring 2010. Sampling time was 4 to 10 days for particles 50, 40, or 30 nm in diameter. Sample air flow was 4 L/min. The sampling setup was arranged so that two samples were obtained for each sampling period almost simultaneously: one containing particles and adsorbed gas-phase compounds and one containing adsorbed gas-phase compounds only. Filters were extracted and analyzed for the presence of selected carboxylic acids, polyols, nitrogen-containing compounds, and aldehydes. The results showed that, in quartz filter samples, gas-phase adsorption may be responsible for as much as 100% of some compound masses. Whether quartz or Teflon, simultaneous collection of gas-phase zero samples is essential during the whole sampling period. The dependence of the adsorption of gas-phase compounds on vapor pressure and the effect of adsorption on the deposited aerosol layer are discussed.


Plasma membrane associated actin in liver of normal and tumour-bearing rats.
A study was made of the association of actin with different plasma membrane fractions from liver of normal rats and from the enlarged liver of rats bearing a Walker 256 carcinoma where a decrease in the state of polymerisation of cytoplasmic actin has been previously observed. As estimated by the DNAase I inhibition assay, actin constituted approx. 7% and 3%, respectively, of the protein of membrane fractions enriched in lateral or bile-canalicular domains, but only trace amounts were found in the sinusoidal fraction. [3H]Cytochalasin B binding indicated the presence of 20 and 13 pmol of high-affinity binding sites per mg protein in lateral and bile-canalicular fractions, but none in the sinusoidal. Kd for cytochalasin B binding was of the order of 1 nM for lateral and bile-canalicular fractions. Polypeptide profiles obtained by SDS/urea/polyacrylamide gel electrophoresis of non-ionic detergent-insoluble residues differed for all three fractions although some proteins, including actin, occurred as major components of both bile-canalicular and lateral regions. Tumour growth had no effect on the actin content, high-affinity cytochalasin B binding or polypeptide profiles of the three membrane fractions.


Downregulation of TAB182 promotes cancer stem-like cell properties and therapeutic resistance in triple-negative breast cancer cells.
TAB182 participates in DNA damage repair and radio-/chemosensitivity regulation in various tumors, but its role in tumorigenesis and therapeutic resistance in breast cancer remains unclear. In the current paper, we observed that triple-negative Breast Cancer (TNBC), a highly aggressive type of breast cancer, exhibits a lower expression of TAB182. TAB182 knockdown stimulates the proliferation, migration, and invasion of TNBC cells. Our study first obtained RNA-seq data to explore the cellular functions mediated by TAB182 at the genome level in TNBC cells. A transcriptome analysis and in vitro experiments enabled us to identify that TAB182 downregulation drives the enhanced properties of cancer stem-like cells (CSCs) in TNBC cells. Furthermore, TAB182 deletion contributes to the resistance of cells to olaparib or cisplatin, which can be rescued by silencing GLI2, a gene downstream of cancer stemness-related signaling pathways. Our results reveal a novel function of TAB182 as a potential negative regulator of cancer stem-like properties and drug sensitivity in TNBC cells, suggesting that TAB182 may be a tumor suppressor gene and is associated with increased therapeutic benefits for TNBC patients.


Long title: Protocol for evaluating a Consultation for Suffering at work in French-speaking Switzerland.
Psychosocial suffering entails human, social and economic costs. In Switzerland, 34.4% of workers report chronic work-related stress. Our medical Consultation for Suffering at Work aims to preserve-or restore-the patient's capacity to act and make decisions after a diagnosis of work-related psychological suffering; it also aims to help employees get back to or remain at work. Our hypothesis is that the dynamic of the consultation itself and adherence to its medical advice are active factors of these results. Understand changes in patients' work and health status 12 months after a Consultation for Suffering at Work. Determine the effects of the consultation on health and working status via identified active factors: the consultation dynamic and the ability to adhere to the consultation's advice. Evaluate the consultation's effects qualitatively. This longitudinal, monocentric study with a quasi-experimental design will include patients consulting between 1 January and 31 December 2018. Changes in patients' work and health status will be analysed using data collected via questionnaires at 0, 3 and 12 months. Qualitative data will be collected via a semi-structured telephone interview 3 months after the consultation. The quantitative part will include 150-170 patients; the qualitative part will include 30. This exploratory research project will provide a better understanding of issues of work-related psychological suffering and effective strategies to support patients. The absence of a control group and the impossibility of applying a randomised controlled design are constraints on this study.


Butanol production from lignocellulosics.
Clostridium spp. produce n-butanol in the acetone/butanol/ethanol process. For sustainable industrial scale butanol production, a number of obstacles need to be addressed including choice of feedstock, the low product yield, toxicity to production strain, multiple-end products and downstream processing of alcohol mixtures. This review describes the use of lignocellulosic feedstocks, bioprocess and metabolic engineering, downstream processing and catalytic refining of n-butanol.


Photo-enhanced electrocatalytic hydrogen evolution reaction coupled semiconductor with plasma in neutral solution.
A gold-ruthenium/zinc oxide nanorod composite was synthesized. The electrochemical catalytic efficiency of the noble metal-semiconductor nanostructure increased by nearly 30% under the irradiation of an external light source. It provides an efficient way of thinking for the design of electrocatalysts with a photoresponse.


Modulation of Slow Desensitization (Tachyphylaxis) of Acid-Sensing Ion Channel (ASIC)1a.
Among the proton-activated channels of the ASIC family, ASIC1a exhibits a specific tachyphylaxis phenomenon in the form of a progressive decrease in the response amplitude during a series of activations. This process is well known, but its mechanism is poorly understood. Here, we demonstrated a partial reversibility of this effect using long-term whole-cell recording of CHO cells transfected with rASIC1a cDNA. Thus, tachyphylaxis represents a slow desensitization of ASIC1a. Prolonged acidifications provided the same recovery from slow desensitization as short acidifications of the same frequency. Slow desensitization and steady-state desensitization are independent processes although the latter attenuates the development of the former. We found that drugs which facilitate ASIC1a activation (e.g., amitriptyline) cause an enhancement of slow desensitization, while inhibition of ASIC1a by 9-aminoacridine attenuates this process. Overall, for a broad variety of exposures, including increased calcium concentration, different pH conditions, and modulating drugs, we found a correlation between their effects on ASIC1a response amplitude and the development of slow desensitization. Thus, our results demonstrate that slow desensitization occurs only when ASIC1a is in the open state.


Cytoskeletal Alteration Is an Early Cellular Response in Pulmonary Epithelium Infected with Aspergillus fumigatus Rather than Scedosporium apiospermum.
Invasive aspergillosis and scedosporiosis are life-threatening fungal infections with similar clinical manifestations in immunocompromised patients. Contrarily, Scedosporium apiospermum is susceptible to some azole derivative but often resistant to amphotericin B. Histopathological examination alone cannot diagnose these two fungal species. Pathogenesis studies could contribute to explore candidate protein markers for new diagnosis and treatment methods leading to a decrease in mortality. In the present study, proteomics was conducted to identify significantly altered proteins in A549 cells infected with or without Aspergillus fumigatus and S. apiospermum as measured at initial invasion. Protein validation was performed with immunogold labelling alongside immunohistochemical techniques in infected A549 cells and lungs from murine models. Further, cytokine production was measured, using the Bio-Plex-Multiplex immunoassay. The cytoskeletal proteins HSPA9, PA2G4, VAT1, PSMA2, PEX1, PTGES3, KRT1, KRT9, CLIP1 and CLEC20A were mainly changed during A. fumigatus infection, while the immunologically activated proteins WNT7A, GAPDH and ANXA2 were principally altered during S. apiospermum infection. These proteins are involved in fungal internalisation and structural destruction leading to pulmonary disorders. Interleukin (IL)-21, IL-1α, IL-22, IL-2, IL-8, IL-12, IL-17A, interferon-γ and tumour necrosis factor-α were upregulated in both aspergillosis and scedosporiosis, although more predominately in the latter, in accordance with chitin synthase-1 and matrix metalloproteinase levels. Our results demonstrated that during invasion, A. fumigatus primarily altered host cellular integrity, whereas S. apiospermum chiefly induced and extensively modulated host immune responses.


Hindcasting the 2017 dispersal of Sargassum algae in the Tropical North Atlantic.
Since 2011, huge amounts of Sargassum algae are detected in the equatorial Atlantic, causing large strandings events on the coasts of the West Indies, Brazil and West Africa. The distribution of this stock shows strong annual and interannual variability, whose drivers are not settled yet. Here we use satellite Sargassum observations from MODIS and currents from an ocean reanalysis to simulate the passive transport of algae in 2017. Wind effect was necessary to fit the observed distribution. Simulations reasonably reproduce the satellite monthly distribution for up to seven months, confirming the prominent role of transport in the distribution cycle. Annual cycle appears as a zonal exchange between eastern (EAR) and western accumulation regions (WAR). EAR is well explained by advection alone, with sharp meridional distribution controlled by converging currents below the inter-tropical Convergence Zone. Instead, WAR is not explained by advection alone, suggesting local growth.


Circulating prostatic acid phosphatase-immunoglobulin complexes in Sjogren's syndrome.
Elevated level of serum prostatic acid phosphatase (PAP) activity in a patient with Sjogren's syndrome was found to be due to the presence of PAP-immunoglobulin complexes in circulation. The patient did not have a prostatic malignancy. The complexes were demonstrated by counter-immunoelectrophoresis, protein A-Sepharose immunoprecipitation and agarose-gel electrophoresis. Free enzyme was not detected in serum, and the activity of the complexed enzyme was probably unaltered on binding with the immunoglobulins.


Separation of bone matrix proteins by calcium-induced precipitation.
It was found that significant precipitation occurred immediately after calcium, at a concentration as low as 2 mM, was added to a desalted solution of EDTA extract of adult bovine femur. The maximal yield of the precipitates was observed at a calcium concentration of 30 mM. These precipitates were dissolved in 0.5 M EDTA, desalted, and characterized by Sepharose CL-6B gel filtration chromatography and high performance gel-exclusion chromatography. Results revealed that the precipitates were enriched in a 40 K protein and a higher molecular weight fraction as compared with the original extract of bone proteins. The 40 K fraction was isolated and identified as osteonectin, as judged from amino acid analysis, electrophoresis, and immunodetection. The supernatant after calcium-induced precipitation predominantly contained osteocalcin and a 50 K protein that was tentatively identified as alpha 2HS protein. Osteonectin was purified from the calcium-induced precipitates from the EDTA extract of bovine bone. By calcium titration using fluorescence spectrometry, the isolated osteonectin showed high affinity to calcium ions with an apparent dissociation constant (K0.5) of 8 x 10(-7) M. Thus, the use of calcium to separate bone proteins, especially osteonectin, was proved to be a useful technique. In addition, calcium-induced precipitation of osteonectin suggested a possible in vivo mechanism via which osteonectin might interact with calcium ions and participate in the initial immobilization of calcium to induce the nucleation of calcification in bone tissue.


Conjunction of semi-mechanistic in vitro-in vivo modeling and population pharmacokinetics as a tool for virtual bioequivalence analysis - a case study for a BCS class II drug.
Virtual bioequivalence trial (VBE) simulations based on (semi)mechanistic in vitro-in vivo (IVIV) modeling have gained a huge interest in the pharmaceutical industry. Sophisticated commercially available software allows modeling variable drug fates in the gastrointestinal tract (GIT). Surprisingly, the between-subject and inter-occasion variability (IOV) of the distribution volumes and clearances are ignored or simplified, despite substantially contributing to varied plasma drug concentrations. The paper describes a novel approach for IVIV-based VBE by using population pharmacokinetics (popPK). The data from two bioequivalence trials with a poorly soluble BCS class II drug were analyzed retrospectively. In the first trial, the test drug product (biobatch 1) did not meet the bioequivalence criteria, but after a reformulation, the second trial succeeded (biobatch 2). The popPK model was developed in the Monolix software (Lixoft SAS, Simulation Plus) based on the originator's plasma concentrations. The modified Noyes-Whitney model was fitted to the results of discriminative biorelevant dissolution tests of the two biobatches and seven other reformulations. Then, the IVIV model was constructed by joining the popPK model with fixed drug disposition parameters, the drug dissolution model, and mechanistic approximation of the GIT transit. It was used to simulate the drug concentrations at different IOV levels of the primary pharmacokinetic parameters and perform the VBE. Estimated VBE success rates for both biobatches well reflected the outcomes of the bioequivalence trials. The predicted 90% confidence intervals for the area under the time-concentration curves were comparable with the observed values, and the 10% IOV allowed the closest approximation to the clinical results. Simulations confirmed that a significantly lower maximum drug concentration for biobatch 1 was responsible for the first clinical trial's failure. In conclusion, the proposed workflow might aid formulation screening in generic drug development.


Trauma exposure and psychometric properties of the life events checklist among adults in South Africa.
Background: Trauma exposure is widespread and linked to chronic physical and mental health conditions including posttraumatic stress disorder. However, there are major gaps in our knowledge of trauma exposure in Africa and on the validity of instruments to assess potentially life-threatening trauma exposure.Objective: The Life Events Checklist for the DSM-5 (LEC-5) is a free, widely used questionnaire to assess traumatic events that can be associated with psychopathology. As part of a case-control study on risk factors for psychosis spectrum disorders, we used the LEC-5 to examine the frequency of traumatic events and to assess the questionnaire's factor structure in South Africa (N = 6,765).Method: The prevalence of traumatic events was measured by individual items on the LEC-5 across the study sample, by case-control status, and by sex. Cumulative trauma burden was calculated by grouping items into 0, 1, 2, 3, and ≥4 traumatic event types. Psychometric properties of the LEC-5 were assessed through exploratory and confirmatory factor analyses.Results: More than 92% of the study sample reported experiencing ≥1 traumatic event; 38.7% reported experiencing ≥4 traumatic event types. The most endorsed item was physical assault (65.0%), followed by assault with a weapon (50.2%). Almost 94% of cases reported ≥1 traumatic event compared to 90.5% of controls (p < .001) and 94% of male participants reported ≥1 traumatic event compared to 89.5% of female participants (p < .001). Exploratory factor analysis revealed a 6-factor model. Confirmatory factor analyses of three models found that a 7-factor model based on the South African Stress and Health survey was the best fit (standardized root mean square residual of 0.024, root mean square error of approximation of 0.029, comparative fit index of 0.910).Conclusion: Participants reported very high exposure to traumatic events. The LEC-5 has good psychometric priorities and is adequate for capturing trauma exposure in South Africa.


Perinatal origins of bronchopulmonary dysplasia-deciphering normal and impaired lung development cell by cell.
Bronchopulmonary dysplasia (BPD) is a multifactorial disease occurring as a consequence of premature birth, as well as antenatal and postnatal injury to the developing lung. BPD morbidity and severity depend on a complex interplay between prenatal and postnatal inflammation, mechanical ventilation, and oxygen therapy as well as associated prematurity-related complications. These initial hits result in ill-explored aberrant immune and reparative response, activation of pro-fibrotic and anti-angiogenic factors, which further perpetuate the injury. Histologically, the disease presents primarily by impaired lung development and an arrest in lung microvascular maturation. Consequently, BPD leads to respiratory complications beyond the neonatal period and may result in premature aging of the lung. While the numerous prenatal and postnatal stimuli contributing to BPD pathogenesis are relatively well known, the specific cell populations driving the injury, as well as underlying mechanisms are still not well understood. Recently, an effort to gain a more detailed insight into the cellular composition of the developing lung and its progenitor populations has unfold. Here, we provide an overview of the current knowledge regarding perinatal origin of BPD and discuss underlying mechanisms, as well as novel approaches to study the perturbed lung development.


[Aspects of the application of models in pharmaceutical research (author's transl)].
Starting from historical examples of the application of models in scientific research, the author treats the present significance of this working method in pharmacy. A short description of various types of models and of the definition of the term "model" is followed by an outline of the major aspects to be considered in using models. The different steps of model elaboration, the requirements to be considered with model elaboration as well as questions concerning the analogy, main properties, goals and limitations of models, and the validity of the results obtained are briefly dealt with and illustrated by examples.


Postoperative pneumonia in the intensive care unit.
Postoperative pneumonia in the surgical patient may occur from atelectasis and inadequate postoperative TV, from ventilator-associated pathophysiology and from aspiration. Each process ultimately elicits the proinflammatory responses from the lung in the effort to eradicate the infectious pathogens. Postoperative pneumonia is best prevented by maintaining a clear understanding of the biologic basis for infection in the lung. Therapy requires aggressive tracheobronchial drainage strategies and antibiotic therapy directed toward the pathogens that are producing the pulmonary inflammatory response.


Urban-rural differences in trajectories of physical activity in Europe from 2002 to 2017.
Here we evaluate the time trends of urban-rural differences in physical activity in the European Union between 2002 and 2017 and the contribution of urbanization on total physical activity changes, using four Eurobarometer surveys (n = 101,373). Trajectories of urban-rural differences in physical activity varied considerably among EU-28 countries. Hierarchical linear regression models revealed that inactivity increased in both urban and rural settings, although it was higher in the latter. Thereby, the urban-rural gap diminished and was even eliminated in some countries. Also, national changes across time were driven by urban places, showing little contribution from urbanization. Our findings suggest that inactivity has risen in Europe regardless of living environments and with regional urbanization development having little influence.


Experimental animal model of surgical wound infection applicable to antibiotic prophylaxis.
Wound infection, defined as accumulation of pus draining spontaneously or after opening of the wound, developed in 19 out of 22 guinea pigs (86%) after intraincisional contamination with 10(7) Escherichia coli plus 10(8) Bacteroides fragilis before wound closure. Antibiotic prophylaxis with gentamicin plus clindamycin significantly reduced the wound sepsis rate from 86% to 29% (p less than 0,001). Pus for bacteriological studies was available in all cases of wound infection but one, and culture always revealed both Escherichia coli and Bacteroides fragilis. The animal model presented employs widely accepted criteria for wound infection, and avoids culture of tissue homogenates.


The effects of type 2 diabetes mellitus on the corneal endothelium and central corneal thickness.
Aim of this study is to evaluate the differences in corneal endothelial cell morphology and corneal thickness in patients with and without type 2 diabetes related to age, disease duration, and HbA1c percentage. This retrospective cross-sectional study included 511 (1022 eyes) type 2 diabetes patients and 900 (1799 eyes) non-diabetic patients. The endothelial cell density (ECD), variation in endothelial cell size (CV), percentage of hexagonal cells, and central corneal thickness (CCT) were analyzed using a noncontact specular microscope and a Pentacam Scheimpflug camera. We also examined the correlation between the corneal parameters and the duration of diabetes. For total ages, the subjects with type 2 diabetes showed significantly lower ECD, hexagonality, higher CV, and thicker CCT than the control group. This difference was more pronounced in patients with long-standing DM (≥ 10 years) and high HbA1c (≥ 7%). When stratified by age group, from the 60 s group, corneal endothelial cell parameters showed a statistically significant difference between DM and control groups. The duration of diabetes was inversely correlated with ECD (r = - 0.167; p = 0.000). These findings suggest that diabetes affects corneal endothelial cell in older age and those with long-standing DM and higher HbA1c. Regular corneal endothelial examinations are required in diabetic patients.


Proton conductivity and proton dynamics in nanocrystalline cellulose functionalized with imidazole.
In the present study, we report the synthesis, electrical and dynamic properties of a new generation bio-based nanocomposite, that is a proton-exchange membrane based on nanocrystalline cellulose (CNC) and imidazole (Im). CNC serves as supporting material and imidazole acts as a proton donor and proton acceptor at the same time. The nanocomposite (1.3 CNC-Im) was synthesized as a film and shows proton conductivity equal to 4.0 × 10-1 S/m at 160 °C in anhydrous conditions. Analysis of impedance measurements and NMR spectra provided some insight into the macroscopic and microscopic processes involved in proton transport in 1.3 CNC-Im. Local processes such as reorientation of imidazole rings and breaking of hydrogen bonds are identified and their activation energies are calculated. The energies of the macroscopic and microscopic proton transport in CNC-Im film are correlated. The percolation model used confirmed the percolation nature of conductivity in cellulose composites with imidazole.


Methionine Restriction Regulates Cognitive Function in High-Fat Diet-Fed Mice: Roles of Diurnal Rhythms of SCFAs Producing- and Inflammation-Related Microbes.
Methionine restriction (MR) is known to potently alleviate inflammation and improve gut microbiome in obese mice. The gut microbiome exhibits diurnal rhythmicity in composition and function, and this, in turn, drives oscillations in host metabolism. High-fat diet (HFD) strongly altered microbiome diurnal rhythmicity, however, the role of microbiome diurnal rhythmicity in mediating the improvement effects of MR on obesity-related metabolic disorders remains unclear. 10-week-old male C57BL/6J mice are fed a low-fat diet or HFD for 4 weeks, followed with a full diet (0.86% methionine, w/w) or a methionine-restricted diet (0.17% methionine, w/w) for 8 weeks. Analyzing microbiome diurnal rhythmicity at six time points, the results show that HFD disrupts the cyclical fluctuations of the gut microbiome in mice. MR partially restores these cyclical fluctuations, which lead to time-specifically enhance the abundance of short-chain fatty acids producing bacteria, increases the acetate and butyric, and dampens the oscillation of inflammation-related Desulfovibrionales and Staphylococcaceae over the course of 1 day. Notably, MR, which protects against systemic inflammation, influences brain function and synaptic plasticity. MR could serve as a potential nutritional intervention for attenuating obesity-induced cognitive impairments by balancing the circadian rhythm in microbiome-gut-brain homeostasis.


Perceived stigma in adults with epilepsy in Sweden and associations with country of birth, socioeconomic status, and mental health.
Stigma contributes to negative health outcomes and amplifies health disparities in epilepsy. This study aimed to investigate associations of perceived stigma with the country of birth and socioeconomic status (SES). This is a cross-sectional questionnaire study. Participants were recruited consecutively from three demographically different neurology outpatient clinics in the southwest of Sweden. Participants responded to a questionnaire concerning their epilepsy, country of birth, mother tongue, and different SES-variables. The Neuro-QOL stigma scale and the Jacoby stigma scale were applied to assess stigma and the Hospital Anxiety and Depression Scale (HADS) and PROMIS Mental Health scale were applied to assess mental health. In total 161 adults with epilepsy were included in the cohort. The median Neuro-QOL stigma score was 48.3, and was higher among foreign-born than in native-born participants (foreign-born in non-European country 52.3, in other European country 49.8, and in native-born 47.0, p = 0.003). Other factors associated with Neuro-QOL were seizure frequency last year (≥2 seizures 52.4 compared to 1 seizure 50.9 and no seizures 44.3, p < 0.001), having had seizures in public (yes 50.9 compared to no 44.7, p = 0.035), HADS depression score ≥11 (57.4 compared to 47.8 for score <11 points, p < 0.001), HADS anxiety score ≥11 (53.5 compared to 46.8 for score <11 points, p < 0.001), and lower PROMIS Mental Health score (42.9 for PROMIS < 40 compared to 54.4 for PROMIS > 60, p < 0.01). A stepwise multiple regression analysis indicated that having had seizures the last year increased the average Neuro-QOL stigma score with 5.89 and appeared as the most determining factor for the Neuro-QOL stigma score among the variables investigated. It is important that the concerns of foreign-born patients are acknowledged and that the focus of seizure control and the detection and treatment of comorbidities are prioritized in the management of epilepsy and perceived stigma.


Characteristics of spinal cord compression in adenocarcinoma of prostate.
Of 611 patients with biopsy-proved adenocarcinoma of the prostate, spinal cord compression developed in 41 (6.7%) at a median interval of twenty-four months after primary diagnosis. Spinal cord involvement most often occurred in the thoracic area, with 95 per cent of patients showing radiographic evidence of osseous vertebral metastasis at the level of cord compression. All lesions but one were located extradurally, and patients with Stage D2 disease, by virtue of bony metastases, were therefore at greatest risk for development of neurologically compressive disease. There was also a significant increase in the incidence of spinal cord involvement among the more poorly differentiated tumors, although tumor histology did not appear to influence the median interval between vertebral metastasis and cord compromise. Survival following spinal cord involvement was relatively poor and unrelated to tumor differentiation. Forty-six per cent of patients survived less than six months and 20 per cent less than two months. The two most noteworthy survivors are alive at thirty and ninety-seven months, the latter after combined treatment for an intradural lesion.


Association of an intragenic microsatellite marker in the CC16 gene with asthma in the Indian population.
The gene for Clara cell secretory protein (CC16) is an ideal candidate for investigating genetic predisposition to asthma because of its role in the airway as an anti-inflammatory molecule, differences in its levels between asthmatics and healthy controls, and its genetic location (11q13). We investigated the association of an SNP (A38G) and an intragenic repeat polymorphism in the CC16 gene with asthma and its associated traits, such as total serum IgE levels, in a case control as well as in a family based study design. A significant association was observed for the microsatellite repeat at the level of alleles and genotypes with asthma (P<0.05) in both the study designs. However, no association was observed for the A38G SNP with asthma. When haplotypes were constructed for these two loci and compared, the haplotype A_18 was found at higher frequency in patients (OR=1.59, 95%CI=1.08, 2.33, P=0.016). Also, in the family based design, a biased transmission was observed for haplotypes from parents to affected offspring (P=0.003). Individually, haplotype A_18 showed preferential transmission (82.6%) to affected offspring (P=0.001), thereby confirming the case-control results. In summary, this is the first study identifying the CC16 gene to be associated with asthma in the Indian population.


Survival of Vibrio parahaemolyticus in oyster shellstock at two different storage temperatures.
In oyster shellstock harvested from Maryland waters Vibrio parahaemolyticus was found to be present, to survive storage for at least 3 weeks at 4 C, and to multiply after being held for 2 to 3 days at 35 C.


Pneumolysin Is Responsible for Differential Gene Expression and Modifications in the Epigenetic Landscape of Primary Monocyte Derived Macrophages.
Epigenetic modifications regulate gene expression in the host response to a diverse range of pathogens. The extent and consequences of epigenetic modification during macrophage responses to Streptococcus pneumoniae, and the role of pneumolysin, a key Streptococcus pneumoniae virulence factor, in influencing these responses, are currently unknown. To investigate this, we infected human monocyte derived macrophages (MDMs) with Streptococcus pneumoniae and addressed whether pneumolysin altered the epigenetic landscape and the associated acute macrophage transcriptional response using a combined transcriptomic and proteomic approach. Transcriptomic analysis identified 503 genes that were differentially expressed in a pneumolysin-dependent manner in these samples. Pathway analysis highlighted the involvement of transcriptional responses to core innate responses to pneumococci including modules associated with metabolic pathways activated in response to infection, oxidative stress responses and NFκB, NOD-like receptor and TNF signalling pathways. Quantitative proteomic analysis confirmed pneumolysin-regulated protein expression, early after bacterial challenge, in representative transcriptional modules associated with innate immune responses. In parallel, quantitative mass spectrometry identified global changes in the relative abundance of histone post translational modifications (PTMs) upon pneumococcal challenge. We identified an increase in the relative abundance of H3K4me1, H4K16ac and a decrease in H3K9me2 and H3K79me2 in a PLY-dependent fashion. We confirmed that pneumolysin blunted early transcriptional responses involving TNF-α and IL-6 expression. Vorinostat, a histone deacetylase inhibitor, similarly downregulated TNF-α production, reprising the pattern observed with pneumolysin. In conclusion, widespread changes in the macrophage transcriptional response are regulated by pneumolysin and are associated with global changes in histone PTMs. Modulating histone PTMs can reverse pneumolysin-associated transcriptional changes influencing innate immune responses, suggesting that epigenetic modification by pneumolysin plays a role in dampening the innate responses to pneumococci.


The use of cerium nitrate-silver sulphadiazine as a topical burns dressing.
Infection remains the major cause of morbidity and mortality following the shock phase in the burn patient. Measures to reduce the risk of wound infection and subsequent sepsis include early excision where possible, and the use of topical antimicrobial creams such as silver sulphadiazine. Studies from the USA and Europe suggest the addition of cerium nitrate to this commonly used agent may improve its efficacy. We present the findings of a pilot study which investigated the action of a commercial preparation of cerium nitrate/silver sulphadiazine mixture (Flammacerium, Duphar B. V. Holland) on 20 patients considered unsuitable for surgery. There were no episodes of cellulitis or septicaemia. Flammacerium was noted to produce an adherent eschar that was easy to shave and which received split skin grafts well.


Ambient particulate matter compositions and increased oxidative stress: Exposure-response analysis among high-level exposed population.
Oxidative stress has been suggested to be one of the key drivers of health impact of particulate matter (PM). More studies on the oxidative potential of PM alone, but fewer studies have comprehensively evaluated the effects of external and internal exposure to PM compositions on oxidative stress in population. To comprehensively investigate the exposure-response relationship between PM and its main compositions with oxidative stress indicators. We conducted a cross-sectional study including 768 participants exposed to particulates. Environmental levels of fine particulate matter (PM2.5), polycyclic aromatic hydrocarbons (PAHs) and metals in PM were measured, and urinary levels of PAHs metabolites and metals were measured as internal dose, respectively. Multivariable linear regression models were used to analyze the correlations of PM exposure and urinary levels of 8-hydroxy-2́'-deoxyguanosine (8-OHdG), and 8-iso-prostaglandin-F2α (8-iso-PGF2α) and malondialdehyde (MDA). The concentration of both PM2.5 and total PAHs was significantly correlated with increased urinary 8-OHdG, 8-iso-PGF2α and MDA levels (all p < 0.05). The levels of 4 essential metals all showed significant exposure-response increase in urinary 8-OHdG in both current and non-current smokers (all p < 0.05); ambient selenium, cobalt and zinc were found to be significantly correlated with urinary 8-iso-PGF2α (p = 0.002, 0.003, 0.01, respectively); only selenium and cobalt were significantly correlated with urinary MDA (p < 0.001, 0.01, respectively). Furthermore, we found each one-unit increase in urinary total OH-PAHs generated a 0.32 increase in urinary 8-OHdG, a 0.22 increase in urinary 8-iso-PGF2α and a 0.19 increase in urinary MDA (all p < 0.001). Furthermore, it was found that the level of 12 urinary metals all showed significant and positive correlations with three oxidative stress biomarkers in all subjects (all p < 0.001). Our systematic molecular epidemiological study showed that particulate matter components could induce increased oxidative stress on DNA and lipid. It may be more important to monitor and control the harmful compositions in PM rather than overall particulate mass.


Individual participant data analysis of two trials on aldosterone blockade in myocardial infarction.
Two recent randomised trials studied the benefit of mineralocorticoid receptor antagonists (MRAs) in ST-segment elevation myocardial infarction (STEMI) irrespective or in absence of heart failure. The studies were both undersized to assess hard clinical endpoints. A pooled analysis was preplanned by the steering committees. We conducted a prespecified meta-analysis of patient-level data of patients with STEMI recruited in two multicentre superiority trials, randomised within 72 hours after symptom onset. Patients were allocated (1:1) to two MRA regimens: (1) an intravenous bolus of potassium canrenoate (200 mg) followed by oral spironolactone (25 mg once daily) versus standard therapy or (2) oral eplerenone (25-50 mg) versus placebo. The primary and key secondary outcomes, all-cause death and the composite of all-cause death or resuscitated sudden death, respectively, were assessed in the intention-to-treat population using a Cox model stratified on the study identifier. Patients were randomly assigned to receive (n=1118) or not the MRA regimen (n=1123). After a median follow-up time of 188 days, the primary and secondary outcomes occurred in 5 (0.4%) and 17 (1.5%) patients (adjusted HR (adjHR) 0.31, 95% CI 0.11 to 0.86, p=0.03) and 6 (0.5%) and 22 (2%) patients (adjHR 0.26, 95% CI 0.10 to 0.65, p=0.004) in the MRA and control groups, respectively. There were also trends towards lower rates of cardiovascular death (p=0.06) and ventricular fibrillation (p=0.08) in the MRA group. Our analysis suggests that compared with standard therapy, MRA regimens are associated with a reduction of death and death or resuscitated sudden death in STEMI.


Pneumothorax as the initial manifestation of idiopathic hypereosinophilic syndrome.
We report a case of hypereosinophilic syndrome in a 47-year-old man who had acute pneumothorax as the initial presentation. Peripheral blood eosinophil count increased continuously over a period of 1 month and was associated with pulmonary changes and appearance of skin lesions on the right chest wall. Idiopathic hypereosinophilic syndrome was confirmed by bone marrow aspiration biopsy and skin lesion biopsy after exclusion of all possible secondary etiologies. The clinical status and chest radiographs showed marked improvement after treatment with corticosteroids.


The impact of early flow and brain oxygen crisis on the outcome of patients with severe traumatic brain injury.
Multimodality monitoring and goal-directed therapy may not prevent blood flow and brain oxygen (Flow/BrOx) crisis. We sought to determine the impact of these events on outcome in patients with severe traumatic brain injury (sTBI). Twenty-four patients with sTBI were treated to maintain intracranial pressure (ICP) less than or equal to 20 mm Hg, cerebral perfusion pressure (CPP) greater than or equal to 60 mm Hg, brain oxygen greater than or equal to 20 mm Hg, and near infrared spectroscopy greater than or equal to 60%. Flow/BrOx crisis events were recorded. The 14-day predicted mortality was compared with actual mortality. Nonsurvivors had a significantly higher number of crisis events nonresponsive to treatment (P < .05). Mortality was 87.5% in patients with greater than or equal to 20 events versus 6.3% in patients with less than 20 events. The predicted mortality was 58%, whereas actual mortality was 33.3% (8/24), yielding a 42% reduction in mortality. A multimodality monitoring and goal-directed therapy may decrease mortality in sTBI. However, Flow/BrOx crisis events still occur and predict a poor outcome.


Cardiovascular risk in subjects with left ventricular concentric remodeling at baseline examination: a meta-analysis.
It is debated whether subjects with concentric remodeling (CR, normal left ventricular mass index (LVMI) and increased relative wall thickness (RWT)) are at higher cardiovascular risk than those with normal geometry (NG, normal LVMI and RWT). The aim of this study was to perform a meta-analysis of studies evaluating cardiovascular events in subjects with CR and NG according to baseline classification. We searched for articles evaluating cardiovascular outcome in subjects with CR compared with those with NG, and reporting adjusted hazard ratio (HR) and 95% confidence interval (CI). Six studies were included in the meta-analysis. The pooled population consisted of 7465 subjects with CR and NG. During the follow-up, they experienced 852 events. When compared with NG, the overall adjusted HR was 1.36 (95% CI 1.03-1.78) for CR, P<0.03. There was some heterogeneity between studies. Subgroup meta-analysis showed that increased cardiovascular risk in subjects with CR was more relevant in studies evaluating hypertensive and Caucasian subjects and reporting both fatal and non-fatal events. Cardiovascular risk is significantly higher in subjects with CR than in those with NG. This aspect is more evident in studies including hypertensive patients and Caucasian populations and reporting global cardiovascular risk.


[Surveillance of Haemophilus Influenzae meningitis in children in France, 2001-2006].
The implementation of Haemophilus influenzae b (Hib) vaccination program in France in 1993 resulted in a rapid and dramatic decrease of the Hib meningitis incidence in children. The aim of our study was to describe the residual Haemophilus influenzae (Hi) meningitis in the French paediatric population between 2001 and 2006. The French Paediatric Infectious Diseases Group set up an active surveillance network to analyze the clinical and biological features of bacterial meningitis. We used these data to retrospectively study the risk factors, signs and symptoms, vaccination status, cerebrospinal fluid analysis, treatment and case fatality rate of Hi meningitis. Among the 2539 cases of bacterial meningitis reported in France between 2001 and 2006, 69 (2.7 %) were due to Hi. A mean number of 11.5 cases of Hi meningitis was reported annually (minimum 6, maximum 16). Among Hi meningitis cases, 36 strains were of serotype b, 8 were capsulated but not b (6 f, 1 e and 1 unknown serotype), 20 strains were not capsulated, and 5 were non studied. The mean age of the children was 30.3 months (median 13.8 months, range 3.3 months to 14.5 years). 41 % of children with Hib meningitis did not received any anti-Hib vaccine and 41 % did not followed the French recommendations for Hib vaccine. Hi meningitis still occurs, and more than half is due to the b serotype. Among Hib cases, 14 % did not recieved any anti-Hib vaccine and 15 % received an incomplete vaccination schedule. Increase of vaccine coverage and use of an earlier booster dose at the age of 12 months could further improve the epidemiology of Hib meningitis. The immunity of children with Hib meningitis should be systematically studied in order to improve the comprehension of the pathophysiology of vaccine failure.


Latex-based thin-layer immunoaffinity chromatography for quantitation of protein analytes.
A rapid immunochromatographic method for qualitative and quantitative analysis of protein antigens is described. The method is based on the "sandwich" assay format using monoclonal antibodies (Mabs) of two distinct specificities. Mabs of one specificity are covalently immobilized to a defined detection zone on a porous membrane while Mabs of the other specificity are covalently coupled to blue latex particles which serve as a label. The sample is mixed with the Mab-coated particles and allowed to react. The mixture is then passed along a porous membrane by capillary action past the Mabs in the detection zone, which will bind the particles which have antigen bound to their surface, giving a blue color within this detection zone with an intensity logarithmetrically proportional to the antigen concentration in the sample. Analysis is complete in less than 10 min, requires a minimum amount of sample (4 microliters), and has a detection limit below the nanomolar range for the antigen we studied, human chorionic gonadotropin.


N,N-dibutylnitrosamine metabolism by rat liver S9 fraction and oxidation by chemical model systems.
Oxidative in vitro metabolism of N,N-dibutylnitrosamine (DBN) by 9000g supernatant fraction (S9 fraction) prepared from rat liver and oxidation of DBN by chemical model systems (Udenfriend and Fenton) were investigated. The products retaining the N-nitroso group of DBN were analyzed by gas-liquid chromatography. Incubation of DBN with the S9 fraction gave hydroxy derivatives of DBN at the omega, omega-1, and omega-2 positions, while the chemical reaction of DBN in the Udenfriend and the Fenton model systems afforded compounds with an oxo group at the same positions. In all cases, omega-1 oxidation was predominant, followed by omega and then by omega-2 oxidations, while omega and omega-1 oxidations were the principal metabolic pathways of DBN in vivo in the rat. The differences in the oxidative transformation of DBN among in vitro enzymatic, non-enzymatic and in vivo systems are discussed.


Anti-GD2 and Anti-CD47 Are Synergistic and Promote Tumor Eradication.
Anti-GD2 and anti-CD47 antibodies show synergy in models of neuroblastoma and osteosarcoma.


A 78 kDa host cell invasion protein of Neospora caninum as a potential vaccine candidate.
Neosporosis is an intracellular protozoan disease caused by Neospora caninum. Until now, there is no effective vaccine to prevent neosporosis. The host cell binding protein has the potential as neosporosis vaccine. In the present study, a T7 phage display library was constructed and screened using Vero cells to obtain host cell binding protein of N. caninum. Two host cell binding proteins, a hypothetical protein of 78 kDa (named as NcP78) homologous to the acylglycerol lipase of Toxoplasma gondii ME49 (XP_002370319.1) and NcGRA7 (known as a dense granules protein that is involved in the invasion of N. caninum to the host cells), were identified. Immune responses induced by recombinant NcP78 and NcGRA7 proteins and their protective efficacies against homologous challenge in BALB/c mice were evaluated respectively. Results showed that recombinant NcP78 and NcGRA7 could elicit both Th1 and Th2 immune responses (with the elevated levels of IgG1 and IgG2a antibody), but predominately a Th2 immune response with a high level of IgG1. The ani-NcP78 and anti-NcGRA7 serum also had inhibitory effects on N. caninum invasion to Vero cells in vitro, which indicated that both NcP78 and NcGRA7 proteins were involved in host cell invasion. Recombinant NcP78 and NcGRA7 could not prolong the survival times and improve the survival rates of dams, but could prolong the survival times and improve the survival rates of offspring significantly. Moreover, the recombinant NcP78 and NcGRA7 could reduce the brain parasite load of dams and offspring. Though these protein vaccines could not effectively alleviate the symptom of abortion, they could increase the number of born offspring significantly, indicating that Nc78 and NcGRA7 recombinant proteins could provide a partial protection against N. caninum infection in mice.


Validation of an Electrochemical Sensor to Detect Cortisol Responses to the Trier Social Stress Test.
Recent advances in sensor technology allow for the detection of salivary cortisol levels in real-time, yet studies are needed to test their reliability in clinically at-risk populations. This study examined whether a new electrochemical sensor reliably detected cortisol patterns, compared to a conventional immunoassay test (i.e., ELISA), among women and men with low and high depressive symptoms who participated in the Trier Social Stress Test (TSST; a laboratory-based stressor). Results demonstrated that women and those with high depressive symptoms showed lower cortisol levels throughout the TSST overall compared to men and those with low depressive symptoms. The cortisol sensor was highly reliable when compared to the ELISA immunoassay in detecting cortisol responses to the TSST for both women and men and for participants with low and high depressive symptoms. These results suggest that the sensor is a promising tool for assessing real-time cortisol responses to laboratory stressors in at-risk populations.


Obesity, metabolic health, and urological disorders in adults: a nationwide population-based study.
We evaluate the risks of various urological disorders that require treatments according to obesity and metabolic health status using a nationwide dataset of the Korean population. 3,969,788 patients who had undergone health examinations were enrolled. Participants were classified as "obese" (O) or "non-obese" (NO) using a BMI cut-off of 25 kg/m2. People who developed ≥ 1 metabolic disease component in the index year were considered "metabolically unhealthy" (MU), while those with none were considered "metabolically healthy" (MH). There were classified into the MHNO, MUNO, MHO, and MUO group. In BPH, chronic renal disease, neurogenic bladder, any medication related to voiding dysfunction, alpha-blocker, and antidiuretics, age and gender-adjusted hazard ratio (HR) was highest in MUO, but higher in MUNO than in MHO. In stress incontinence, prostate surgery, and 5alpha-reductase, HR increased in the order of MUNO, MHO, and MUO. In prostatitis, anti-incontinence surgery, and cystocele repair, HR was higher in MHO than MUNO and MUO. In cystitis, cystostomy, and anticholinergics, HR was higher in MUNO and MUO than MHO. In conclusion, obesity and metabolic health were individually or collaboratively involved in urological disorders related to voiding dysfunction. Metabolic healthy obesity needs to be distinguished in the diagnosis and treatment of urological disorders.


A rare variant of the mtDNA HVS1 sequence in the hairs of Napoléon's family.
This paper describes the finding of a rare variant in the sequence of the hypervariable segment (HVS1) of mitochondrial (mtDNA) extracted from two preserved hairs, authenticated as belonging to the French Emperor Napoléon I (Napoléon Bonaparte). This rare variant is a mutation that changes the base C to T at position 16,184 (16184C→T), and it constitutes the only mutation found in this HVS1 sequence. This mutation is rare, because it was not found in a reference database (P < 0.05). In a personal database (M. Pala) comprising 37,000 different sequences, the 16184C→T mutation was found in only three samples, thus in this database the mutation frequency was 0.00008%. This mutation 16184C→T was also the only variant found subsequently in the HVS1 sequences of mtDNAs extracted from Napoléon's mother (Letizia) and from his youngest sister (Caroline), confirming that this mutation is maternally inherited. This 16184C→T variant could be used for genetic verification to authenticate any doubtful material and determine whether it should indeed be attributed to Napoléon.


Binding parameters of antibodies reacting with multivalent antigens: functional affinity or pseudo-affinity.
For antibodies binding bivalently to multivalent antigens, including the cell surface, it has been common to determine the 'functional affinity', even though it is clear that the equations describing true affinity only apply to monovalent interactions. We here summarize evidence that 'functional affinities' are not valid, because they do not accurately describe the interactions that occur. A basic discrepancy is that dissociation, in practice, is generally not a first-order reaction. A more useful approach to compare antibodies is to determine the kinetics of dissociation over a long time period, until near-complete dissociation has occurred.