Organic walled microfossils in wet peperites from the early Cretaceous Paraná-Etendeka volcanism of Brazil.

Large igneous provinces (LIPs) are major magmatic events that have a significant impact on the global environment and the biosphere, for example as triggers of mass extinctions. LIPs provide an excellent sedimentological and geochemical record of short but intense periods of geological activity in the past, but their contribution towards understanding ancient life is much more restricted due to the destructive nature of their igneous origin. Here, we provide the first paleontological evidence for organic walled microfossils extracted from wet peperites from the Early Cretaceous Paraná-Etendeka intertrappean deposits of the Paraná basin in Brazil. Wet peperites are a volcaniclastic rock formed by the interaction of lava and subaqueous sediments.The Paraná-Etendeka was formed during the Valanginian (ca. 132 Ma) as a continental flood basalt in present day South America and Namibia, and released enormous amounts of carbon dioxide, sulfur dioxide, methane and hydrogen fluoride into the atmosphere. The organic walled microfossils recovered from the Paraná-Etendeka peperites include pollen grains, spores, acritarchs, and other remains of unidentifiable organic matter. In addition to the peperites, organic walled microfossils were also found in heterolithic sandstones and interpillow sandstones. Our findings represent the first insight into the biodiversity of the Paraná Basin during the Early Cretaceous during a period of intense magmatism, and the microfossil assemblages corroborate a regional paleoclimatic transition from arid to more humid conditions that were likely induced by the volcanic activity. We corroborate the potential of wet peperite rocks as a valuable source of paleobiological data and emphasize the importance of sampling volcaniclastic units that have been traditionally considered with lower fossiliferous potential due to their igneous origin.

A leukotriene-dependent spleen-liver axis drives TNF production in systemic inflammation.

Production of the proinflammatory cytokine tumor necrosis factor (TNF) must be precisely regulated for effective host immunity without the induction of collateral tissue damage. Here, we showed that TNF production was driven by a spleen-liver axis in a rat model of systemic inflammation induced by bacterial lipopolysaccharide (LPS). Analysis of cytokine expression and secretion in combination with splenectomy and hepatectomy revealed that the spleen generated not only TNF but also factors that enhanced TNF production by the liver, the latter of which accounted for nearly half of the TNF secreted into the circulation. Using mass spectrometry-based lipidomics, we identified leukotriene B4 (LTB4) as a candidate blood-borne messenger in this spleen-liver axis. LTB4 was essential for spleen-liver communication in vivo, as well as for humoral signaling between splenic macrophages and Kupffer cells in vitro. LPS stimulated the splenic macrophages to secrete LTB4, which primed Kupffer cells to secrete more TNF in response to LPS in a manner dependent on LTB4 receptors. These findings provide a framework to understand how systemic inflammation can be regulated at the level of interorgan communication.

Trifocal toric intra-ocular lens implantation in pediatric traumatic cataract.

PURPOSE: To present the case of a 6-year-old child who presented with a traumatic cataract and was treated with trifocal toric intra-ocular lens implantation. OBSERVATIONS: The child's uncorrected distance visual acuity improved from +0.7 logMAR preoperatively to 0.00 logMAR after cataract surgery, with spectacle independence and no reported side effects. CONCLUSIONS AND IMPORTANCE: In carefully selected pediatric patients with traumatic cataracts, trifocal toric intra-ocular lenses may offer some benefit over standard monofocal lenses.

DMH-CBD, a cannabidiol analog with reduced cytotoxicity, inhibits TNF production by targeting NF-kB activity dependent on A2A receptor.

Cannabidiol (CBD) is a natural compound with psychoactive therapeutic properties well described. Conversely, the immunological effects of CBD are still poorly explored. In this study, the potential anti-inflammatory effects and underlying mechanisms of CBD and its analog Dimethyl-Heptyl-Cannabidiol (DMH-CBD) were investigated using RAW 264.7 macrophages. CBD and DMH-CBD suppressed LPS-induced TNF production and NF-kB activity in a concentration-dependent manner. Both compounds reduced the NF-kB activity in a µM concentration range: CBD (IC50 = 15 µM) and DMH-CBD (IC50 = 38 µM). However, the concentrations of CBD that mediated NF-kB inhibition were similar to those that cause cytotoxicity (LC50 = 58 µM). Differently, DMH-CBD inhibited the NF-kB activation without cytotoxic effects at the same concentrations, although it provokes cytotoxicity at long-term exposure. The inhibitory action of the DMH-CBD on NF-kB activity was not related to the reduction in IkBα degradation or either p65 (NF-kB) translocation to the nucleus, although it decreased p38 MAP kinase phosphorylation. Additionally, 8-(3-Chlorostyryl) caffeine (CSC), an A2A antagonist, reversed the effect of DMH-CBD on NF-kB activity in a concentration-dependent manner. Collectively, our results demonstrated that CBD reduces NF-kB activity at concentrations intimately associated with those that cause cell death, whereas DMH-CBD decreases NF-kB activity at non-toxic concentrations in an A2A receptor dependent-manner.

Pharmacokinetic properties, in vitro metabolism and plasma protein binding of govaniadine an alkaloid isolated from Corydalis govaniana Wall.

Govaniadine (GOV) is an alkaloid isolated from Corydalis govaniana Wall. It has been reported to show a different number of biological activities including anti-urease, leishmanicidal and antinociceptive. The present study aims to characterize the GOV in vitro metabolism after incubation with rat and human liver microsomes (RLM and HLM, respectively) and to evaluate its pharmacokinetic properties. The identification of GOV metabolites was conducted by different mass analyzers: a micrOTOF II-ESI-ToF Bruker Daltonics® and an amaZon-SL ion trap (IT) Bruker Daltonics®. For the pharmacokinetic study of GOV in rats after intravenous administration, a LC-MS/MS method was developed and applied to. The analyses were performed using an Acquity UPLC® coupled to an Acquity TQD detector equipped with an ESI interface. The liver microsomal incubation resulted in new O-demethylated, di-hydroxylated and mono-hydroxylated compounds. Regarding the method validation, the calibration curve was linear over the concentration range of 2.5-3150.0ngmL-1, with a lower limit of quantitation (LLOQ) of 2.5ngmL-1. This method was successfully applied to a pharmacokinetic study. The profile was best fitted to a two-compartment model, the first phase with a high distribution rate constant (α) 0.139±0.086min-1, reflected by the short distribution half-life (t1/2α) 9.2±8.9min and the later one, with an elimination half-life (t1/2ß) 55.1±37.9min. The main plasma protein binding was 96.1%. This is a first report in this field and it will be useful for further development of govaniadine as a drug candidate.

Metabolic profile and safety of piperlongumine.

Piperlongumine (PPL), a natural plant product, has been extensively studied in cancer treatment going up on clinical trials. Since the first report related to its use on cancer research (in 2011) around 80 papers have been published in less than 10 years, but a gap still remaining. There are no metabolism studies of PPL in human organism. For the lack of a better view, here, the CYP450 in vitro oxidation of PPL was described for the first time. In addition, the enzymatic kinetic data, the predicted in vivo parameters, the produced metabolites, the phenotyping study and possible piperlongumine-drug interactions in vivo is presented.

MicroRNA-29b Overexpression Decreases Extracellular Matrix mRNA and Protein Production in Human Corneal Endothelial Cells.

PURPOSE: MicroRNAs are small noncoding RNAs that regulate gene expression at the posttranscriptional level. We reported that levels of microRNA (miR)-29 family are decreased in corneas of patients with Fuchs endothelial corneal dystrophy (FECD). The miR-29 family regulates the production of extracellular matrix (ECM) proteins. Accumulation of ECM proteins in Descemet membrane is an important pathologic change in FECD. In this study, we transfected miR-29b into human corneal endothelial cells and tissues and evaluated ECM protein expression levels. METHODS: An immortalized Fuchs human corneal endothelial cell line (iFECD) was established by infection of corneal endothelial cells from patients with FECD with hTERT lentivirus. MiR-29b was transfected into iFECD, and the expression levels of ECMs collagen type 1 alpha 1 (COL1A1), collagen type 4 alpha 1 (COL4A1), and laminin gamma 1 (LAMC1) were evaluated with quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) and Western blot. Expression level of LAMC1 protein in miR-29b-transfected donor corneal endothelium was also evaluated by Western blot. RESULTS: Compared with control, miR-29b expression level after transfection of iFECD was increased to 335.6% (±91.0%), and ECM expression levels were significantly decreased. Compared with control, qRT-PCR demonstrated reduction of ECM to the following levels: COL1A1: 1.9% (±0.4%); COL4A1: 7.1% (±1.7%); and LAMC1: 21.5% (±2.7%). Western blot showed reduced protein expression: COL1A1: 4.8% (±3.2%); COL4A1: 42.5% (±25.0%); and LAMC1: 44.8% (±3.1%). In miR-29b-transfected corneal tissue, LAMC1 protein expression level was decreased to 14.4% (±20.5%). CONCLUSIONS: Overexpression of miR-29b decreased ECM protein production in human corneal endothelial cells. Thus, miR-29 replacement therapy might be a new treatment strategy for FECD aimed at reducing pathologic production of ECM proteins in Descemet membrane.

Assessment of a Novel Corneal-Shaping Device With Simultaneous Corneal Collagen Cross-Linking Using a Porcine Eye Model.

PURPOSE: Corneal collagen cross-linking (CXL) alone cannot substantially improve refractive errors. We designed a novel corneal-shaping device, showing that a prototype applied during CXL can alter corneal curvature to a greater extent than CXL alone in a porcine eye model. METHODS: The device prototype was made with flat, UV-transmissible material. Enucleated porcine eyes were deepithelialized and perfused. Preexperimental and postexperimental corneal curvature (K) measurements and Anterior Segment Optical Coherence Tomography imaging were performed. A conventional CXL protocol was followed. Six experiments of 6 treatment groups in duplicate were performed (n = 12/group): (A) no CXL + no device, (B) no CXL + 30 minutes device, (C) 10 minutes, CXL + no device, (D) 10 minutes, CXL + device, (E) 30 minutes, CXL + no device, and (F) 30 minutes, CXL + device. RESULTS: There was a significant decrease (P < 0.001) in corneal curvature after 30 minutes. CXL between group F (-1.54 ± 0.90 D) and groups E (+0.34 ± 0.53 D), B (+0.69 ± 1.02 D), or A (+0.22 ± 1.24 D). The demarcation line depth was greater in group F (34% ± 4%) than group D (30% ± 4%, P < 0.05). Groups D (-2.3% ± 21%) and F (0.24% ± 15%) had less change in corneal thickness than in control groups A (15% ± 12%, P < 0.05) and E (12% ± 11%, P < 0.05). CONCLUSIONS: Application of our corneal-shaping device during CXL significantly decreased corneal curvature compared with controls. Anterior Segment Optical Coherence Tomography imaging showed comparable extent of cross-linking in eyes treated ± device. This demonstrates that our novel device used during CXL may further promote corneal strengthening and refractive correction in patients.

Risk factors for eye bank preparation failure of Descemet membrane endothelial keratoplasty tissue.

PURPOSE: To assess the results of a single eye bank preparing a high volume of Descemet membrane endothelial keratoplasty (DMEK) tissues using multiple technicians to provide an overview of the experience and to identify possible risk factors for DMEK preparation failure. DESIGN: Cross-sectional study. METHODS: setting: Lions VisionGift and Wilmer Eye Institute at Johns Hopkins Hospital. STUDY POPULATION: All 563 corneal tissues processed by technicians at Lions VisionGift for DMEK between October 2011 and May 2014 inclusive. OBSERVATION PROCEDURES: Tissues were divided into 2 groups: DMEK preparation success and DMEK preparation failure. MAIN OUTCOME MEASURES: We compared donor characteristics, including past medical history. RESULTS: The overall tissue preparation failure rate was 5.2%. Univariate analysis showed diabetes mellitus (P = .000028) and its duration (P = .023), hypertension (P = .021), and hyperlipidemia or obesity (P = .0004) were more common in the failure group. Multivariate analysis showed diabetes mellitus (P = .0001) and hyperlipidemia or obesity (P = .0142) were more common in the failure group. Elimination of tissues from donors either with diabetes or with hyperlipidemia or obesity reduced the failure rate from 5.2% to 2.2%. Trends toward lower failure rates occurring with increased technician experience also were found. CONCLUSIONS: Our work showed that tissues from donors with diabetes mellitus (especially with longer disease duration) and hyperlipidemia or obesity were associated with higher failure rates in DMEK preparation. Elimination of tissues from donors either with diabetes mellitus or with hyperlipidemia or obesity reduced the failure rate. In addition, our data may provide useful initial guidelines and benchmark values for eye banks seeking to establish and maintain DMEK programs.

Variability in Oculus Pentacam tomographer measurements in patients with keratoconus.

PURPOSE: The aim of this study was to determine intertest variability of measurements taken by the Pentacam tomographer in eyes with keratoconus. METHODS: Three Pentacam measurements with 1-minute intervals were taken from 101 eyes (81 keratoconus and 20 controls) from 57 patients (47 keratoconus and 10 controls) by the same examiner and under the same conditions. Keratoconus eyes were divided into 3 subgroups based on topographic astigmatism (TA). Variability in astigmatism, mean keratometry, central corneal thickness, and fixation values were compared within the patient subgroups versus normal controls. RESULTS: Variability of all studied parameters (astigmatism, mean keratometry, central corneal thickness, and fixation values) increased with increasing TA in patients with keratoconus. CONCLUSIONS: Pentacam measurements show increased variability in patients with keratoconus. The degree of variability correlates with TA.

Retained host Descemet membrane (Auto-DMET) during conversion of deep anterior lamellar keratoplasty to penetrating keratoplasty: a case report.

PURPOSE: The aim of this study was to relate a case of deep anterior lamellar keratoplasty (DALK) converted to penetrating keratoplasty in which the host central Descemet membrane (DM) with a large perforation was left attached to the host cornea. METHODS: This is a case report of a 34-year-old man who underwent an attempted DALK for keratoconus in his left eye, which became complicated with a large rupture in DM during dissection. The host DM was left in place, the donor cornea with DM intact was sutured onto the host bed, and air was injected into the anterior chamber. The patient was monitored by biomicroscopy, pachymetry, topography, anterior segment optical coherence tomography, and specular microscopy. RESULTS: The postoperative course resulted in improved uncorrected visual acuity, best spectacle-corrected visual acuity, and topography. Corneal edema was observed in the host cornea peripheral to the graft. Three months after the surgery, the endothelial cell density was reduced by 63% compared with the preoperative donor cell density. CONCLUSIONS: Leaving the host DM during conversion of DALK to penetrating keratoplasty was uneventful in this case, although corneal edema was observed in the area overlying the host cornea. It is possible that the retained DM could provide additional autologous endothelial cells to prolong graft survival.

Leishmanicidal evaluation of tetrahydroprotoberberine and spirocyclic erythrina-alkaloids.

Leishmaniasis is one of the World's most problematic diseases in developing countries. Traditional medicines to treat leishmaniasis have serious side effects, as well as significant parasite resistance problems. In this work, two alkaloids 1 and 2 were obtained from Corydalis govaniana Wall and seven alkaloids 3-9, were obtained from Erythrina verna. The structures of the compounds were confirmed by mass spectrometry and 1D- and 2D-NMR spectroscopy. The leishmanicidal activity of compounds 1-9 against Leishmania amazonensis was tested on promastigote forms and cytotoxicity against J774 (macrophage cell line) was assessed in vitro. Compound 1 showed potent activity (IC50 = 0.18 µg/mL), compared with the standard amphotericin B (IC50 = 0.20 µg/mL). The spirocyclic erythrina-alkaloids showed lower leishmanicidal activity than dibenzoquinolizine type alkaloids.

Flavodoxin overexpression confers tolerance to oxidative stress in beneficial soil bacteria and improves survival in the presence of the herbicides paraquat and atrazine.

AIM: To determine whether expression of a cyanobacterial flavodoxin in soil bacteria of agronomic interest confers protection against the widely used herbicides paraquat and atrazine. METHODS AND RESULTS: The model bacterium Escherichia coli, the symbiotic nitrogen-fixing bacterium Ensifer meliloti and the plant growth-promoting rhizobacterium Pseudomonas fluorescens Aur6 were transformed with expression vectors containing the flavodoxin gene of Anabaena variabilis. Expression of the cyanobacterial protein was confirmed by Western blot. Bacterial tolerance to oxidative stress was tested in solid medium supplemented with hydrogen peroxide, paraquat or atrazine. In all three bacterial strains, flavodoxin expression enhanced tolerance to the oxidative stress provoked by hydrogen peroxide and by the reactive oxygen species-inducing herbicides, witnessed by the enhanced survival of the transformed bacteria in the presence of these oxidizing agents. CONCLUSIONS: Flavodoxin overexpression in beneficial soil bacteria confers tolerance to oxidative stress and improves their survival in the presence of the herbicides paraquat and atrazine. Flavodoxin could be considered as a general antioxidant resource to face oxidative challenges in different micro-organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of plant growth-promoting rhizobacteria or nitrogen-fixing bacteria with enhanced tolerance to oxidative stress in contaminated soils is of significant agronomic interest. The enhanced tolerance of flavodoxin-expressing bacteria to atrazine and paraquat points to potential applications in herbicide-treated soils.

Extended lateral approach for intra-articular calcaneal fractures: an inverse relationship between surgeon experience and wound complications.

The current reference standard for the treatment of displaced intra-articular calcaneal fractures is open reduction and internal fixation using an extended lateral approach. In the present retrospective study, we evaluated the results of a consecutive series of patients treated in the same fashion from June 2005 to September 2011 using a subcuticular single-layer closure technique. We also determined the risk factors for the development of wound complications and the rate of wound complications. Also, we assessed which patient, fracture, and surgical characteristics affected these complications. During the 75-month study period, we operated on 53 displaced intra-articular calcaneal fractures in 50 patients using the extended lateral approach. The incision was closed using the subcuticular technique in 49 cases (92.45%). In the subcuticular closure group 2 (4.1%) deep infections and 2 (4.1%) superficial wound complications (1 dehiscence and 1 infection) occurred. Wound edge or flap necrosis was not encountered. The use of bone-void filler and the experience of the surgical team were significantly (p < .001 and p = .026, respectively) associated with the occurrence of wound complications. The subcuticular single-layer suture technique is a suitable closure technique in the treatment of displaced intra-articular calcaneal fractures. It was associated with a low complication rate combined with the extended lateral approach. The effect of bone void fillers on the incidence of complications should receive more attention in future research. The association between wound complications and the experience level of the surgical team supports the need for centralization of this complex injury.

Humeral shaft fractures: retrospective results of non-operative and operative treatment of 186 patients.

BACKGROUND: Humeral shaft fractures account for 1-3% of all fractures and 20% of the fractures involving the humerus. The aim of the current study was to compare the outcome after operative and non-operative treatment of humeral shaft fractures, by comparing the time to radiological union and the rates of delayed union and complications. METHODS: All patients aged 16 years or over treated for a humeral shaft fracture during a 5-year period were included in this retrospective analysis; periprosthetic and pathological fractures were excluded. Radiographs and medical charts were retrieved and reviewed in order to collect data on fracture classification, time to radiographic consolidation and the occurrence of adverse events. RESULTS: A total of 186 patients were included; 91 were treated non-operatively and 95 were treated operatively. Mean age was 58.7 ± 1.5 years and 57.0% were female. In 83.3% of the patients, only the humerus was affected. A fall from standing height was the most common cause of the fracture (72.0%). Consolidation time varied from a median of 11-28 weeks. The rate of radial nerve palsy in both groups was similar: 8.8% versus 9.5%. In 5.3% of the operatively treated patients, the palsy resulted from the operation. Likewise, delayed union rates were similar in both groups: 18.7% following non-operative treatment versus 18.9% following surgery. CONCLUSION: The data indicated that consolidation time and complication rates were similar after operative and non-operative treatment. A prospective randomised clinical trial comparing non-operative with operative treatment is needed in order to examine other aspects of outcome, meaning shoulder and elbow function, postoperative infection rates, trauma-related quality of life and patient satisfaction.

Effects of salt stress and rhizobial inoculation on growth and nitrogen fixation of three peanut cultivars.

Increasing soil salinity represents a major constraint for agriculture in arid and semi-arid lands, where mineral nitrogen (N) deficiency is also a frequent characteristic of soils. Biological N fixation by legumes may constitute a sustainable alternative to chemical fertilisation in salinity-affected areas, provided that adapted cultivars and inoculants are available. Here, the performance of three peanut cultivars nodulated with two different rhizobial strains that differ in their salt tolerance was evaluated under moderately saline water irrigation and compared with that of N-fertilised plants. Shoot weight was used as an indicator of yield. Under non-saline conditions, higher yields were obtained using N fertilisation rather than inoculation for all the varieties tested. However, under salt stress, the yield of inoculated plants became comparable to that of N-fertilised plants, with minor differences depending on the peanut cultivar and rhizobial strain. Our results indicate that N fixation might represent an economical, competitive and environmentally friendly choice with respect to mineral N fertilisation for peanut cultivation under moderate saline conditions.

The management of acute distal tibio-fibular syndesmotic injuries: results of a nationwide survey.

INTRODUCTION: Ankle fractures are one of the most frequently encountered musculoskeletal injuries, and 10% of patients have a concomitant distal tibiofibular syndesmotic disruption necessitating surgical repair. A national survey was conducted to gain more insight into the current approaches in the management of syndesmotic injuries in the Netherlands. MATERIALS AND METHODS: A postal survey was sent to one or two staff members of the trauma and orthopaedic surgery departments in each of the 86 hospitals in the Netherlands. Questions concerned the pre-, per- and postoperative strategies and the different ideas on the type, number and placement of the syndesmotic screw. RESULTS: A total of 85.2% of the trauma surgeons and 61.9% of the orthopaedic surgeons responded (representing 87% of all hospitals). Syndesmotic injury was judged mainly using the 'Hook test'. Syndesmotic injuries in a Weber-B ankle fracture were treated with one screw in 81.2% of cases and in Maisonneuve injuries mainly with two screws. The 3.5-mm screw was used most frequently over three cortices at 2.1-4.0cm above the tibial plafond. Removal of the syndesmotic screw was routinely done by 87.0% of surgeons, mostly between 6 and 8 weeks. Of all respondents, 62.3% showed interest in participating in a randomised controlled trial comparing standard removal with removal on indication. CONCLUSION: Compared with previous surveys our survey is more complete, has the highest response rate and has almost national coverage. Most individual items reviewed compare well with current literature, except for the routine removal of the syndesmotic screw, which might not be encouraged from a literature point of view. For this reason, the results of the current survey will be used in the development of a multicentre randomised controlled trial comparing the functional outcome in routine removal of the syndesmotic screw compared with removal on indication.

Hemi-Castaing ligamentoplasty for the treatment of chronic lateral ankle instability: a retrospective assessment of outcome.

PURPOSE: In the treatment of chronic ankle instability, most non-anatomical reconstructions use the peroneus brevis tendon. This, however, sacrifices the natural ankle stabilising properties of the peroneus brevis muscle. The aim of this study was to evaluate the functional outcome of patients treated with a hemi-Castaing procedure, which uses only half the peroneus brevis tendon. METHODS: We performed a retrospective cohort study of patients who underwent hemi-Castaing ligamentoplasty for chronic lateral ankle instability between 1993 and 2010, with a minimum of one year follow-up. Patients were sent a postal questionnaire comprising five validated outcome measures: Olerud-Molander Ankle Score (OMAS), Karlsson Ankle Functional Score (KAFS), Tegner Activity Level Score (pre-injury, prior to surgery, at follow-up), visual analog scale on pain (VAS) and the Short Form 36 (SF-36). RESULTS: Twenty patients completed the questionnaire on functional outcome. The OMAS showed good to excellent outcome in 80% and the KAFS in 65%, the Tegner Score improved from surgery but did not reach pre-injury levels, the VAS on pain was 1 of 10 and the SF-36 returned to normal compared with the average population. CONCLUSIONS: Even though most patients were satisfied with the results, outcome at long-term follow-up was less favourable compared with the literature on anatomical reconstructions. In accordance with the literature, we therefore conclude that the initial surgical treatment of chronic lateral ankle instability should be an anatomical repair with augmentation (i.e. the Broström-Gould technique) and the non-anatomical repair should be reserved for unsuccessful cases after anatomical repair or in cases where no adequate ligament remnants are available for reconstruction.