To present a method, alternative to penetrating keratoplasty, for the restoration of impaired corneal clarity with anterior stromal scarring following long-standing corneal graft failure.A 48-year old female who had previously underwent Descemet stripping automated endothelial keratoplasty (DSAEK) for the treatment of pseudophakic bullous keratopathy, presented with long-standing corneal oedema and anterior corneal scarring. A significant improvement in corrected distance visual acuity was demonstrated, as corneal clarity was restored following graft exchange and phototherapeutic keratectomy (PTK).The combination of corneal graft exchange and phototherapeutic keratectomy may represent an effective therapeutic option for long-standing corneal oedema with concomitant anterior corneal scarring after failure of a DSAEK graft. Descemet stripping automated endothelial keratoplasty (DSAEK) has become the modality of choice for the treatment of corneal oedema arising from corneal endothelial diseases including Fuchs endothelial dystrophy and pseudophakic bullous keratopathy .In cases of DSAEK graft failure or rejection, corneal graft exchange could be attempted. Nevertheless, when corneal oedema is left to ensue, the subsequent chronic corneal decompensation may result in anterior corneal fibrosis. The aforementioned complication limits the final visual outcome of a new DSAEK procedure and, thus, may alter the surgeon’s plan in favour of penetrating keratoplasty (PK).A 48-year old female sought medical consultation due to decreased vision of the left eye. Her past medical history included no necessity for spectacle correction at a younger age and phacoemulsification surgery of the left eye 4 years ago. Following cataract surgery, no improvement of her visual acuity was noticed due to pseudophakic bullous keratopathy. Therefore, the patient underwent DSAEK procedure in the left eye six months post cataract extraction. Restoration of good visual acuity following DSAEK that lasted for the next year was noticed; upon the aforementioned period the patient reported gradual deterioration of vision. Gradual visual impairment was attributed to graft failure.The ophthalmic examination of the left eye revealed uncorrected distance visual acuity HM (hand movement) that could not be improved with spectacles. Slit-lamp biomicroscopy of the left eye showed corneal oedema accompanied by anterior corneal fibrosis.We proceeded with a second Descemet stripping endothelial keratoplasty  in order to replace the non-functional graft with a healthy one. The procedure resulted in complete resolution of corneal oedema within the first postoperative month Fig . NeverthIn the first postoperative month, anterior corneal fibrosis resolved Fig  and the Corneal decompensation arising from a non-functional graft or from endothelial diseases such as Fuchs endothelial dystrophy  and pseuIn our case, failure of DSAEK graft led to chronic corneal oedema with a central opacity that involved the anterior stroma. We decided against PK in an attempt to achieve not only satisfying post-keratoplasty visual acuity, as well as to obviate the increased morbidity of a full-thickness corneal graft. A two-step procedure that included DSAEK graft exchange and phototherapeutic keratectomy with adjunctive MMC was performed. To date, 6 months following PTK, no recurrence of the anterior corneal fibrosis has been observed.Manual debridement of the fibrosis would be an alternative in case of a superficial scar. However, in the case of scar extension into the anterior stroma, manual peeling may give rise to deep corneal defects that may, in turn, lead to uneven healing and an unpredictable visual outcome . In addiIn conclusion, DSAEK graft failure prompts consideration for timely restoration of subsequent corneal oedema, as chronic corneal decompensation can result in anterior corneal fibrosis. In the adverse event of graft failure with anterior corneal scarring, combined graft exchange and PTK should be considered in an effort to spare the patient from the increased morbidity of a PK graft and to achieve a greater visual outcome.


The book combines fields which experience increasing interest in recent years: optofluidics, microfluidics and fighting multiple drug resistance (MDR) seen as part of infectious diseases domain. Ther eported results are of interest for life sciences, environment quality control and biomedical basic and applied research. Biomedical specialists, chemists, physicists, public health experts and even outer space researchers may be among target readers, as well as students engaged in these fields.The book shows a convincing and useful connection between optofluidics and MDR studies. It is distributed along 18 chapters written by 40 authors from 9 countries and 12 laboratories.A set of chapters informs readers about selected non-antibiotic medicines which are exposed to UV pulsed laser beams for generating photoproducts with enhanced properties in fighting MDR. Such parent compounds are phenothiazines, quinazolines and hydantoin derivatives which do not have normally significant effects on bacteria or tumour tissues, but after being exposed to laser radiation in water (chosen as a biocompatible liquid) solutions, generate photoproducts with individual or synergistic effects on biological targets.The book shows the most recent results in the action of exposed chlorpromazine and thioridazine on Gram-positive and Gram-negative bacteria and their enhanced antibacterial and antibiofilm activity.Complementary data about the effects of two cytostatics, methotrexate and 5-Fluorouracil, exposed to optical radiation on eye pseudotumours are synthesised; showing that mixtures of photoproducts generated from them have higher anti-inflammatory effects then their parent compounds.These data are correlated with reports about microfluidic properties for microdroplets serving as vehicles for the transport of medicines to targets.


Mol. Biol. Cell. (2015)]we used the well-established yeast [PSI+]/Sup35 and[+PIN¬]/Rnq1 prion models to show thatautophagy prevents sporadic prion formation. Importantly, we found thatspermidine, a polyamine that has been used to increase autophagic flux, acts asa protective agent which prevents spontaneous prion formation. Prions are self-perpetuating amyloid protein aggregates which underlie variousneurodegenerative diseases in mammals. The molecular basis underlying theirconversion from a normally soluble protein into the prion form remains largelyunknown. Studies aimed at uncovering these mechanism(s) are therefore essentialif we are to develop effective therapeutic strategies to counteract thesedisease-causing entities. Autophagy is a cellular degradation system which haspredominantly been considered as a non-selective bulk degradation process whichrecycles macromolecules in response to starvation conditions. We now know thatautophagy also serves as a protein quality control mechanism which selectivelydegrades protein aggregates and damaged organelles. These are commonlyaccumulated in various neurodegenerative disorders including prion diseases. Inour recent study [Speldewinde  Oxidativelydamaged Sup35 was found to accumulate in mutants lacking core components of theautophagy pathway, and this was found to correlate with an increased frequency ofde novo [PSI+] prion formation. Weshowed that growth under anaerobic conditions in the absence of molecular oxygenprevented the accumulation of oxidized Sup35 and abrogated the high frequency of[PSI+] formation in an autophagy mutant. This suggeststhat autophagy normally functions to clear oxidatively damaged proteins prior to theirconversion to the prion form. A protective role for autophagy in preventing denovo prion formation was further confirmed by showing that increasingautophagic flux by treatment with spermidine abrogates the formation of prions inmutants which normally show high rates of de novo prion formation. Thisimportant new finding strongly implicates autophagy as a defense system which protectsagainst oxidative damage of the non-prion form of a protein. This is an importanttrigger for the formation of the heritable prion conformation, an event that has alsobeen implicated in the formation of mammalian prions.The molecular basis by which prions arise spontaneously is poorly understood. Our dataindicate that oxidative protein damage to Sup35, which is a known trigger for Our study highlights the potential use for autophagy-inducing agents such as spermidinein the prevention of the very early stages of spontaneous prion formation i.e.effectively acting as a prion prevention agent . By improving the clearance ofdamaged/misfolded proteins, there is less possibility for these abnormal proteins toaccumulate and to act as nucleation sites catalyzing the aggregation of other damagedproteins. Polyamines such as spermidine are polycations which play multiple roles incell growth, proliferation and longevity. Their beneficial effects in prolonginglifespan are thought to be mediated by increasing autophagic flux. Spermidine inhibitshistone acetylases and the resulting alterations in the acetylproteome increases thetranscription of different autophagy-related genes . More work will be requiredto determine whether the abrogation of prion formation by spermidine solely depends onincreasing autophagic flux, or whether spermidine additionally promotes other stressprotective pathways. For example, spermidine supplementation has been linked withincreased stress tolerance including heat and oxidative stress, which is not onlymediated by increasing autophagic flux. Whether spermidine modulates the expression ofother stress responsive genes, such as heat shock and antioxidant genes which are knownto influence protein misfolding and prion formation, has not been fully established.Hence, spermidine may amelioarte prion fomation via multiple mechnisms including theinduction of autophagy and other stress-related pathways. PSI+]-versions ofwild-type and atg1 mutant strains. Following short-term induction ofthe Sup35NM-GFP fusion construct, fluorescent foci can be detected due to thecoalescence of newly made Sup35NM-GFP with pre-existing Sup35 aggregates. FluorescentSup35 aggregates are normally visible in approximately 70% of[PSI+] cells examined . We found that spermidinetreatment reduced this number such that visible Sup35 aggregates are only detected inapproximately 25% of cells. This did not occur in an atg1 mutantconfirming the requirement for an active autophagy pathway to clear aggregates inresponse to spermidine treatment . This experiment suggests that increasingautophagic flux via spermidine treatment, not only promotes the removal of smallermisfolded/oxidized Sup35 proteins, but can also promote the removal of larger molecularweight Sup35 aggregates which are already formed within cells. It should be emphasizedthat visible Sup35-GFP aggregates do not necessarily correspond to the number of trueheritable [PSI+] aggregates in cells and more work will berequired to examine whether spermidine treatment can really cure cells of the[PSI+] prion. This may be unlikely though, since thepresence of only a few low molecular weight Sup35 propagons will be inherited bydaughter cells resulting in [PSI+] prion transmission.However spermidine may well be beneficial in the treatment of other non-heritable andamorphous protein aggregate diseases. Recent data from our lab, which was not included in the original pulication, demonstratesthat spermidine treatment can also promote the clearance of Sup35 aggregates from cellsin an autophagy-dependent manner . We used a Sup35NM-GFP fusion construct tovisualize Sup35 aggregate formation in [Polyamines, such as spermidine, are present in millimolar quantities within alleukaryotic cells. They play essential roles in a multitude of cellular processes relatedto cell growth, proliferation and metabolism. Spermidine is a naturally occurringpolyamine and rich dietary sources include soy products, legumes, corn, and whole graincereals. The cellular levels of polyamines, such as spermidine, decline with age andhave been linked to lifespan and age-related disorders. Supplementation of spermidineinto dietary regimes may therefore have added benefits for cellular health and healthyageing. Relative to other established pharmacological inducers of autophagy, such asrapamycin and resveratrol, spermidine can be readily obtained from dietary sources anddoes not exhibit deleterious side effects. This places spermidine as a promisingtherapeutic agent for the prevention and amelioration of protein homeostasis and relatedaggregation diseases.


Parvalbumin inhibitory interneurons (PVIs) are crucial for maintaining proper excitatory/inhibitory balance and high-frequency neuronal synchronization. Their activity supports critical developmental trajectories, sensory and cognitive processing, and social behavior. Despite heterogeneity in the etiology across schizophrenia and autism spectrum disorder, PVI circuits are altered in these psychiatric disorders. Identifying mechanism(s) underlying PVI deficits is essential to establish treatments targeting in particular cognition. Based on our previous publications and new data, we propose oxidative stress as a common pathological mechanism leading to PVI impairment in schizophrenia and some forms of autism.Using immunohistochemistry technique and confocal imaging analysis, we assessed the relationship between oxidative stress  and PVI and their perineuronal net (PNN) in twelve established animal models relevant to autism  and schizophrenia  KO, GRIN2A KO, Gclm KO) with or without additional insult , methylazoxymethanol acetate developmental rodent model (MAM) and poly:IC).When PVI deficits in the anterior cingulate cortex were found in these animal models carrying genetic and/or environmental risks relevant to diverse etiological aspects of these disorders, oxidative stress was always present. Specifically, oxidative stress was negatively correlated with the integrity of PVIs and the extracellular perineuronal net enwrapping these interneurons. Oxidative stress may result from dysregulation of systems typically affected in schizophrenia, including glutamatergic, dopaminergic, immune, and antioxidant signaling. As convergent endpoint, redox dysregulation has successfully been targeted to protect PVIs with antioxidants/redox regulators across several animal models . D-serine, an allosteric modulator of brain NMDA receptor also protected PVIs and PNN against oxidative stress in SR KO mice.In view of the fact that the established pathophysiological processes dopamine excess, immune dysregulation and NMDA receptor hypofunction could all induce oxidative stress and are potentiated by additional oxidative insults, this mechanism could be central to damage of the highly metabolically active PVIs and the PNN surrounding them. Antioxidant systems are therefore potential therapeutic targets, assuming that redox regulators could be applied early, during environmental impacts, long before the clinical emergence of the disease.


Oral delivery is the most accepted and economical route for drug administration and leads to substantial reduction in dosing frequency. However, this route still remains a challenge for the pharmaceutical industry due to poorly soluble and permeable drugs leading to poor oral bioavailability. Incorporating bioactives into nanostructured lipid carriers (NLCs) has helped in boosting their therapeutic functionality and prolonged release from these carrier systems thus providing improved pharmacokinetic parameters. The present review provides an overview of noteworthy studies reporting impending benefits of NLCs in oral delivery and highlights recent advancements for developing engineered NLCs either by conjugating polymers over their surface or modifying their charge to overcome the mucosal barrier of GI tract for active transport across intestinal membrane. Lay abstract: Oral administration of drugs is considered to be a convenient route; however, various drugs that are insoluble in water or unable to permeate across GI tract membrane cannot be delivered by this route. To deliver them effectively, various lipid carriers have been widely explored by researchers. Lipid carriers encapsulate drug inside them and deliver them effectively via the oral route. Also, encapsulation of drug protects them from degradation inside GI tract and safely delivers them to the site of action. This review summarizes application of lipid carriers, in other words, nanostructured lipid carriers, in eradicating these problems, with suitable examples. It is also highly preferred for chronically administered agents, such as anti-tumor, antidiabetic and antihypertensive agents. Drug candidates, which are stable in gastric environment, possess adequate hydrophilic lipophilic balance to cross the intestinal epithelium membrane with the absence of the significant GI tract (GIT), irritation and toxicity signs are ideal candidates for oral delivery . Unfortu effects ,2. Thereet al. extensively worked on biocompatible and biodegradable solid lipids to develop solid lipid nanoparticles (SLNs) in early 1990 and thus resolved the stability and toxicity issues associated with the conventional lipid-based delivery systems [For bioavailability enhancement, the researchers have attempted various approaches to overcome the challenges associated with oral delivery, such as nanosizing of the drug molecules, salt formation, prodrug synthesis and encapsulation of drugs in nanosized carriers, such as polymeric micelles, nanoparticles, liposomes, emulsions, etc. ,3–4. Var systems . The dru systems . Modifie systems .NLCs possess unique characteristics and are formulated using a combination of solid and liquid lipids where less ordered structures are produced, which offer the firmer inclusion of the drug molecules within the matrix during the shelf life ,15. It hin vitro release patterns in both SLNs and NLCs were similar but NLCs displayed high-percent cumulative drug release in comparison to SLNs in 55 h. A lesser mobility of drug in SLNs  in comparison to NLCs (disordered arrangement) was responsible for the slower release of drug. Differential scanning calorimetric analysis showed decreased recrystallization index of NLCs in comparison to solid lipids and physical mixture of solid lipid and liquid lipid favoring the formation of disordered arrangement and reduced capacity of solid lipids to recrystallize suggesting their higher long-term stability. The results of in vivo studies also suggested NLCs to be superior as they exhibited 2.29-fold increase in oral bioavailability when administered to mice. Similar results were also noted with lovastatin-loaded NLCs. Study of partitioning behavior of lovastatin in pure solid lipid and mixtures of solid lipid and liquid lipid also depicted higher partitioning of drug in the lipid phase consisting of a mixture of solid lipid (Precirol® ATO 5) and liquid lipid  thus suggesting that higher solubility of drug was favored by the presence of liquid lipid [As mentioned previously NLCs overcome the disadvantages associated with SLNs, in other words, they provide higher drug loading, faster release rate and storage stability due to a use of blend of solid lipid and liquid lipid in their formulations. id lipid . Anotherid lipid  and dompid lipid  SLNs andNLCs are generally composed of solid lipid, liquid lipid, emulsifiers and water. They are fabricated using various methods, such as emulsification-sonication, solvent injection, high-pressure homogenization, microemulsion and solvent diffusion techniques that have been reviewed elsewhere ,13,22. Tet al. observed that small sized NLCs (200 nm) showed higher AUC values in comparison to higher sized NLCs (600 nm) when administered orally to rats, this was attributed to their higher mucoadhesion capability in the body [Literature reveals that composition and processing parameters employed for fabricating NLCs play a vital role in defining their particle size, percent entrapment efficiency and drug release profile and have been manipulated by various formulation scientists to obtain desired characteristics. Tailoring these properties can be advantageous for obtaining high therapeutic levels of drug in blood plasma. For instance, Muchow the body .et al. revealed that higher solubility of miconazole in lipid provided its better encapsulation in NLCs. In this study, the authors screened various solid lipids  and liquid lipids for formulating miconazole-loaded NLCs. Gelucire 43/01 was selected as solid lipid because it presented highest solubility for miconazole. Among two liquid lipids, in other words, Migilol® 812 and Capryol® PGMC, short listed on the basis of solubility study, capryol PGMC was employed for the fabrication of NLCs as it had higher compatibility with solid lipid selected [et al. carried out 1H NMR analysis to determine any interaction between lipid and bicalutamide as it contains functional groups capable of accepting and donating protons that may interact with the lipids during the preparation of NLCs leading to incompatibility [Solid lipid and liquid lipids, the major constituents of NLCs, ideally should be biocompatible, biodegradable, chemically stable and not possess any toxic effects. The selection of lipids for formulating NLCs is usually done on the basis of solubility of the drug in the lipids as it has a direct impact on drug entrapment and loading efficiency of NLCs . For insselected . Similarselected . Hydrophselected . Kumbhartibility . FTIR antibility .et al. [et al. also reported that the particle size of NLCs was significantly affected by the presence of liquid lipid. Pure solid lipids generated higher sized particles, whereas continuous increment in liquid lipid content led to decrease in particle size due to decrease in viscosity and higher molecular mobility [et al. wherein the continuous addition of Labrafil M 1944CS to Compritol 888 ATO led to the formation of small-sized NLCs. Better entrapment efficiency was also obtained with the addition of liquid lipid due to enhanced solubility of drug facilitated by addition of liquid lipid [Amount of liquid lipid is another parameter that plays a significant role in tailoring particle size and release rate as it leads to reduction in viscosity and surface tension of the system that provides smaller sized NLCs, which in turn provides high surface area, promoting higher percent cumulative drug release as demonstrated by Tiwari et al. . Chen etmobility . A similid lipid .Various lipids/oils obtained from plants are also gaining popularity in the pharmaceutical field for formulating lipid nanoparticles. For instance, tripterine was encapsulated in lipid derived from an edible plant, in other words, broccoli by solvent diffusion method and enhanced oral bioavailability was achieved in comparison to Precirol ATO 5-based lipid nanoparticles and drug suspension suggesting the applicability of plant-based lipids . Also, sAmount of total lipid matrix is also known to affect particle size and entrapment efficiency of NLCs, in other words, an enhancement in both characteristics was observed with increasing the lipid matrix. The higher viscosity of the system is held responsible for growth in particle size, whereas reduction in drug escaping tendency due to higher lipid content results in higher entrapment efficiency. Thus, selection of optimal lipid content is crucial while formulating NLCs with desired properties .in vitro release study. Improved characteristics of myversol system resulted in higher bioavailability of lovastatin-loaded NLCs in orally administered rats  to determine the in vivo fate and exact absorption mechanism employing water quenching fluorescent probes demonstrated no evidence of absorption of intact nanoparticles through the intestinal membrane after oral administration [in vivo studies on engineered NLCs are not satisfactory and there is necessity of detailed future studies to determine effects of composition, physiochemical properties and surface modification on the fate of lipid nanoparticles.During GIT passage, NLCs circumventing digestion process can be either conveyed to the portal blood via paracellular route bypassing metabolism due to enterocyte enzymes or can be captured by M cells of Payer's patches delivering NLCs to the lymphatic system . Extensistration . Howeverstration ,35. Alsostration . HoweverOwing to the poor bioavailability of most of the developed drugs, various research groups have worked on designing suitable oral delivery systems for such drugs. NLCs have gained high recognition as suggested by various research reports published so far for drugs, such as isoliquiritigenin, fenofibrate, baicalin, etc. ,43,47. VAppropriate solubility of drug at the absorption site is necessary for achieving acceptable oral bioavailability. However, more than 40% of new chemical entities coming out from combinatorial screening programs are poorly aqueous soluble, which is a subject of major concern to formulation scientists . NLCs haet al. developed NLCs of thymoquinone for oral delivery using high pressure homogenization technique as an aid to improve its gastroprotective activity. Rats pretreated with thymoquinone-loaded NLCs developed significantly reduced areas of ulcers in stomach in comparison to rats pretreated with plain NLCs against ethanol-induced ulcers via higher modulation of pH of gastric contents [Lercanidipine HCl is an antihypertensive agent that suffers from low solubility and presents poor oral bioavailability (10%). Lercanidipine HCl-loaded NLCs led to the enlargement in surface area, which resulted in an improved release rate to a receptor compartment (pH 7.4) across the rat stomach membrane in comparison to a drug suspension suggesting a higher dissolution rate of the drug. Moreover, drug-loaded NLCs showed superior antihypertensive activity for 24 h in comparison to drug suspension . Repaglicontents .50 values for drug-loaded NLCs were decreased up to five times in comparison to free drug due to the small size of NLCs that were highly absorbed by cells and released high concentration of drug within cells suggesting the applicability of this delivery vehicle for poorly soluble drugs.NLCs have also shown improvement in the pharmacokinetic profile of flavanoid drug; isoliquiritigenin having multiple health benefits, but presenting diminished therapeutic efficacy of its oral administration in suspension form due to its lipophillic nature. In this study, NLCs composed of glyceryl monostearate (solid lipid); Miglyol 812 (liquid lipid) and Tween 80 (surfactant) were developed. Higher diffusion of NLCs across the unstirred water layer of intestine epithelial cells facilitated by the presence of hydrophilic surfactant led to augmented values of AUC in comparison to drug solution. Moreover, the small size of NLCs facilitated bioadhesion to intestine membrane, which lead to higher absorption of NLCs . Also etAn orally ingested substance is exposed to varying conditions in different sections of GIT, which can adversely affect the stability of ingested bioactives. For instance, being a hydrophilic and acid labile compound, ifosfamide degrades at stomach pH and has a limited access to cancerous cells. Also, sintering of this compound on storage decreases its dissolution rate, which also presents problems in its delivery. More recently, ifosfamide was encapsulated inside NLCs having chitosan coating over their surface to provide sustained release of the drug. Formulated NLCs showed only 13% release at pH 1.2 for 6 h, suggesting suitability of NLCs in delivering ifosfamide effectively inside the body . PreviouIn vitro antioxidant study also suggested higher potential of lutein-loaded NLCs [The bioactives are susceptible to different environmental conditions and it is mandatory to protect them from such harsh conditions during storage to provide them better shelf life. For example, lipophillic nutraceuticals, such as luetin, β-carotene or quercetin are obligatory due to their multiple health benefits, but their low aqueous solubility and poor chemical stability during storage presents problems in their oral absorption. To trounce these difficulties, researchers have explored the capability of NLCs for accommodating great amounts of such poorly water soluble bioactives and prevent their premature decomposition . β-carotded NLCs . Thus, pet al. reported an increase in the plasma half life of biochanin A from 6 to 21 h after single oral administration in rats when encapsulated in NLCs [max (1.918-fold) and AUC values (3.2-fold) in comparison to a drug suspension after single administration in Wistar rats [Appropriately designed NLCs may act as a delivery vehicle capable of protecting drugs from premature degradation during their transport across GIT due to first pass metabolism. NLCs interact with bile salt in GIT to form mixed micelles, which undergo selective lymphatic uptake, thus bypassing the liver . Also, t in NLCs . In anot in NLCs . Flow cytar rats . In a sitar rats .1/2 (2 h), which restricts its clinical use due to need of frequent dosing. After encapsulation of simvastatin in NLCs, AUC values were improved up to 4.87-fold and t1/2 was almost doubled in comparison to a drug suspension [et al. combined two novel strategies in a single system, in other words, vinpocetine was incorporated in cyclodextrin complexes that were further incorporated in NLCs matrix and showed 39% higher in vitro release at pH 6.8 and 7.4 and superior bioavailability in male rabbits (92%) in comparison to conventional NLCs [Simvastatin is a drug of choice for treatment of hypercholesterolemia and is available as an immediate release formulation. It is poorly water soluble and highly metabolized in intestinal gut and liver and exhibits very short tspension . Furtherspension . In an anal NLCs .Ex vivo studies showed that formulation having 247-nm particle size (NLCs B) avoided P-gp efflux as it entered by both pathways, whereas other formulations entered only by caveolae pathway (NLCs A and NLCs C) as depicted in P-gp, is a 170 kDa protein belonging to the ATP-binding cassette family and is a well-studied protein, which mainly transports large, hydrophobic, cationic or electrically neutral molecules that can have remarkably dissimilar structures . P-gp isWhen a drug is administered orally, it enters the systemic circulation and gets distributed in different tissues of the body because of its physiochemical properties, which leads to decreased activity of drug than expected and in turn shows drug-associated toxicity. Therefore, it is a prerequisite for a delivery vehicle to deliver the drug at the site of action and at the same time be safe and should also minimize the adverse effects of drug for the effective treatment of any disease. NLCs have helped in improving treatment of various diseases via encapsulating drugs inside them and actively targeting disease area thus minimizing its potential toxicity. For example, tamoxifen is used to treat cancer and its use is accompanied by systemic and hepatic toxicity. Tamoxifen was integrated into NLCs composed of glyceryl monostearate of stearic acid and labrafil WL 2609 BS. On oral administration of 1.5-mg/kg dose of tamoxifen-loaded NLCs, the mice displayed slower clearance and did not show any increased levels of hepatotoxicity markers, whereas their levels were highly elevated with the same dose of tamoxifen suspension and its marketed formulation suggesting the potential of NLCs in circumventing its toxicity .et al. carried out in vitro cytotoxicity studies on Caco-2 cells for validating potential of NLCs in reducing toxicity of tripterine. Encapsulation of tripterine decreased cell killing with an IC50 value of 2.11 μg/ml, whereas cells treated with an equivalent concentration of drug solution exhibited higher cytotoxicity. Significant reduction in intestinal toxicity of the drug was also observed, this could be attributed to the controlled release of tripterine after its encapsulation in NLCs [Oral delivery of tripterine is also associated with adverse affects on GIT and kidneys. Chen  in NLCs . Triptol in NLCs .max, mean residence time and AUC values in comparison to drug solution and marketed oral granules demonstrating the high therapeutic potential of NLCs in addition to overcoming its side effects.Metabolites of few drugs also lead to adverse affects in the body, which necessitates the development of sustained release system for them. For instance, metabolites of montelukast are known to cause hepatotoxicity during its conventional oral therapy. It is documented that long-chain lipids assure lymphatic uptake of NLCs and high drug concentration in mesenteric lymph nodes from NLCs composed with long-chain lipid, in other words, glycerylmonostearate of stearin has been observed . TherefoNLCs have also been employed for effective delivery of drugs in the oral cavity, especially with increasing their residence time in oral mucosa. The mucoadhesive properties of positively charged NLCs have promoted their interaction with negatively charged oral mucosa, thus providing optimal residence in the oral cavity. These systems have been studied to be employed for treatment of various bacterial and fungal infections of the oral cavity. However, considering the lower viscosity of NLCs dispersion, these systems are needed to be incorporated in semisolid preparations, such as hydrogels, to permit their better application in oral mucosa. Recently, miconazole-loaded NLCs have been studied to provide local delivery and controlled release to oral mucosa, which otherwise presents difficulty due to its poor solubility. In this study, the antifungal activity of drug-loaded NLCs was found to be more pronounced in comparison to its marketed formulation having 17-fold higher dose in comparison to NLCs . SimilarIn vitro studies revealed that thiomer (cysteine) conjugation onto the surface of docetaxel-loaded NLCs using postinsertion technique led to higher mucoadhesion (81.6%) with mucin in comparison to unconjugated NLCs (51.9%) [in situ intestinal perfusion investigation. Better pharmacokinetic parameters of modified NLCs in Sprague Dawley rats were also in concordance with results of intestinal perfusion study suggesting beneficial effects of engineered NLCs.The tight epithelial cells of the GIT are covered with a hydrophilic and negatively charged protective layer of mucus that restricts the passage of foreign particles across GIT. However, researchers have exploited mucus as a beneficial tool to increase plasma concentration and therapeutic efficacy of drugs via formulating engineered nanoparticles having the capability to anchorage with mucus. Binding of nanoparticles to mucus increases their residence time in GIT, which allows passive transport of drugs leading to their increased absorption. Two different approaches employing physiochemical properties of mucus have been documented to provide mucoadhesion property to nanoparticles A & B. Fi (51.9%) . Also, sIn vitro drug permeability study across mucus secreting cells (Caco-2/HT 29 MTX) depicted that coating of dextran-protamine to the surface of NLCs could remarkably (twofold) enhance the drug delivery across the cell in comparison to uncoated NLCs [It has been documented that customary rejuvenation of mucus layer by a turnover process hampers bioadhesion process . Therefoted NLCs .et al. observed prolonged-plasma concentration (24 h) in case of docetaxel-loaded PEGylated NLCs in comparison to drug solution, which was very low within 12 h postoral administration to rats as PEGylation prevented opsonin binding to intact NLCs in systemic circulation thus avoiding their macrophage uptake [et al. also incorporated an anticancer drug biochanin A in PEG-NLCs with the aim of achieving improved oral bioavailability [in vitro cytotoxicity study carried out over MCF-7 cell lines highlighted the superior performance of biochanin A loaded PEG-NLCs in comparison to drug suspension. Further, oral bioavailability of etoposide-loaded DSPE-PEG-NLCs in rats was also significantly improved when compared with conventional NLCs and drug suspension, which was attributed to their higher uptake due to opening of tight junctions of intestinal membrane by DSPE-PEG and reduced RES uptake [Hydrophilic moieties, such as poly(ethylene glycol) (PEG), also have the capability to penetrate the aqueous mucus layer of the GIT D. PEG coe uptake . Wang etlability . ResultsS uptake . Thus, Pin vivo. Thus, engineered NLCs have been used as a delivery vehicle for targeting drugs to specific tissues along with improving its pharmacokinetic profile  encapsulating montelukast [ζ-potential is one of the fundamental parameters for assessing colloidal stability, in other words, values of ζ-potential above ±30 mV are the foremost requirement for the electrostatic stabilization due to repulsion between particles . In addi profile E. For ex . NLCs also showed concentration-dependent cytotoxic effects and hemolytic activity [Safety of nanomaterials employed for therapeutic or diagnostic purposes is one of the prime concerns in the modern arena. Numerous  systems . Reportee nature . Similare nature  or any ae nature . An oraldelivery . In a moactivity . In an iactivity . Thus, iex vivo potency, these candidates suffer from poor oral bioavailability and other issues, such as poor permeability, high first pass metabolism, toxicity concerns and instability in GIT. Increase in pubmed data since last decade indicated huge interest of researchers toward lipid-based nanoparticles. Successful NLCs based dermal products (Cutanova Nanorepair Q10 and FloraGlo®) introduced on the market have also directed formulation scientists to evaluate their potential in other routes. Researchers have explored the capability of these lipid nanoparticles for accommodating great amounts of bioactives, preventing their premature decomposition and enhancing their oral bioavailability. The various surfactants employed in NLCs formulations can overcome P-gp efflux and could also enhance permeability of drugs across the intestinal membrane by causing disruption in the membrane. Better pharmacokinetic parameters of drug-loaded NLCs obtained during different studies suggested that NLCs can serve as promising tool for enhancing therapeutic efficacy of drugs and also providing controlled release of encapsulated drugs. Different engineered NLCs investigated for further boosting their potential suggested that tuning of physical characteristics of NLCs via changing their composition/method of fabrication could provide fruitful results. Although the exact mechanism underlying behind enhanced absorption by NLCs needs to be addressed and solubility of drugs in different lipids needs to be determined as drugs show limited solubility in various lipids limiting their dose. Thus, further detailed in vivo studies are mandatory for taking them to clinics and considering the increased research on this system, it can be concluded that the NLCs based system intended for oral delivery will definitely reach clinical studies in near future.The poorly water soluble nature of drugs, resulting from drug discovery processes remains a major concern for pharmaceutical scientists despite their constant hard work. Besides their high Nanostructured lipid carriers (NLCs) are promising oral delivery vehicle due to their higher stability during storage and ease of scalability without any requirement of sterile conditions.These systems are capable in entraping both hydrophilic and lipophilic drugs.Biocompatible and biodegradable lipids employed in their fabrication minimize toxicity issues.Various studies have been reported based on NLCs for overcoming solubility, permeability, stability and toxicity issues of bioactives.Different engineered NLCs for generating mucus adhesive, mucus-penetrating particles or for better targeting have been explored.Peptidic ligands are also finding wider applicability in oral delivery as they facilitate active transport of the drugs loaded inside NLCs across the intestinal membrane.Better pharmacokinetic parameters of drug-loaded NLCs have been obtained during different studies.in vivo studies addressing their mechanism underlying enhanced oral bioavailability are needed for bringing these systems in clinics.Further, significant


This study characterized regions of DNA which remained unassembled by either PacBio and Illumina sequencing technologies for seven bacterial genomes. Two genomes were manually finished using bioinformatics and PCR/Sanger sequencing approaches and regions not assembled by automated software were analyzed. Gaps present within Illumina assemblies mostly correspond to repetitive DNA regions such as multiple rRNA operon sequences. PacBio gap sequences were evaluated for several properties such as GC content, read coverage, gap length, ability to form strong secondary structures, and corresponding annotations. Our hypothesis that strong secondary DNA structures blocked DNA polymerases and contributed to gap sequences was not accepted. PacBio assemblies had few limitations overall and gaps were explained as cumulative effect of lower than average sequence coverage and repetitive sequences at contig termini. An important aspect of the present study is the compilation of biological features that interfered with assembly and included active transposons, multiple plasmid sequences, phage DNA integration, and large sequence duplication. Our targeted genome finishing approach and systematic evaluation of the unassembled DNA will be useful for others looking to close, finish, and polish microbial genome sequences. Since the first Next-Generation Sequencing (NGS) platform was released by 454 Life science  is available  and algorithm developments have facilitated improved genome assemblies. Progress in next-generation sequencing platforms, metrics, and performances has been reviewed  PacBio sequence coverage  and PacBio RS-II platforms. Clostridium pasteurianum ATCC 6013   to remove bases having PHRED quality score <30 and any reads shorter than 20 bp. Adapter trimming and filtering of raw PacBio data was performed through SMRT analysis software to obtain “filtered subreads” with default parameters  . Positional preference was determined using PerPlot and PerScan tools  and B. cellulosolvens DSM 2933 (48X). Post-trimming and filtering statistics for Illumina and PacBio data including the number of reads, average read lengths and genome coverage and total bases are summarized in Tables Illumina sequence coverage for each genome is >200X, sufficient to derive high-quality draft genome assemblies  values. Mfold analysis of PacBio gap sequences revealed the potential to form small stem-loop structures but large and/or strong secondary structural loops that might interfere with DNA polymerase and result in low sequence coverage were not identified. Significant differences were not observed between minimum free energies and secondary structures of PacBio gaps and 20 randomly selected regions from the AD2 and DSM 2933 genomes Table . In addiFor further characterization, we analyzed 1 kb DNA sequences flanking PacBio gaps  from three near-finished genomes in this study. A self-blast was performed using 1 kb regions as a query against the entire genome using Geneious software with default parameters. The grade score from Geneious software  for the top blast hits for gap termini regions are described in Table C. thermocellum LQRI (LQRI), and P. fermentans DSM 17108 genomes  of 16S rRNA  genome assemblies. Short reads from Illumina technology have limited power to resolve longer repetitive regions , but random errors in the PacBio platform can be corrected by using high (>100x) sequence coverage and/or Illumina data. However, uniform sequence coverage across the entire genome is not guaranteed and low coverage regions are prone to base-call errors. Assembly polishing is a crucial step to obtain accurate consensus sequence and facilitate downstream applications. Two assembly base-call correction algorithms applied in this study are Quiver (correction using PacBio reads) and Pilon (correction using Illumina reads) while iCORN . Therefore, for future projects, the application of circlator software followed by a careful inspection of the trimmed regions is recommended.Another important aspect of finished genome sequences is an accurate representation of a circular chromosome. Automatically finished assemblies generated through HGAP often have (duplicated) overlapping ends which need to be trimmed off for the final assembly. This could be achieved using the circulator (Hunt et al., de novo microbial genome assemblies. Our datasets and analyses will aid future efforts to better understand and overcome unassembled DNA from PacBio assemblies.In this study, we present an effective manual finishing approach targeted toward near-finished microbial genome assemblies. The importance of genome polishing steps is demonstrated through its positive influence on gene calling accuracy and improved protein coding potential, which will be useful to others looking to improve long-read assemblies. Assessment of Illumina gaps confirmed previous findings that repetitive rRNA operons are major contributors to fragmented short-read assemblies. For PacBio assemblies, our initial hypothesis that structural features of DNA might affect the PacBio sequence coverage leading to assembly gap was not accepted. However, we demonstrated that certain biological features such as presence of active transposons, plasmid sequences, and phage integration are possible reasons for assembly fragmentation. Additionally, DNA regions flanking the PacBio gap sequences showed high degrees of similarity with other loci and are likely contributors to incomplete PacBio assemblies in this dataset. The PacBio gap sequences in this study are attributed to a cumulative effect of various aspects of repetitive DNA content and biological features for specific genomes. Despite a few limitations, long reads from third-generation sequencing, in this case from the PacBio platform, are particularly advantageous for generating SU designed the study, performed, and contributed to all the experiments and analyses and wrote the manuscript draft; DK extracted genomic DNA, performed Illumina sequencing, and assisted with PCR and Sanger sequencing; RH contributed to study design and edited the manuscript; SB contributed to study design, assisted with draft writing, and editing. All authors reviewed and approved the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In a policy forum, Daniel Schar and colleagues discuss the need for surveillance of antimicrobial consumption in animals in low- and middle-income countries and propose the establishment of antimicrobial consumption monitoring systems. Antimicrobial use in low- and middle-income country (LMIC) food animal production sectors is accelerating, commensurate with expanded intensive production to meet rapidly increasing demand for animal-source nutrition.However, antimicrobial consumption in animal production contexts of LMICs remains largely undocumented, limiting the ability to establish and monitor progress toward achieving consumption targets.We propose the establishment of antimicrobial consumption monitoring systems in a phased manner that will be responsive and adaptive to LMIC contexts, while directing a path toward incremental enhancement of monitoring structures.This phased approach enables implementation of systems yielding standardized, globally comparable antimicrobial consumption data, which could inform policies to optimize antimicrobial usage in food animal production.The approach should be complemented by efforts to strengthen animal production systems, eliminate medically important antimicrobial growth promoters, and reduce reliance upon prophylactic antimicrobial use.Global antimicrobial consumption in terrestrial and aquatic food animal production is accelerating, associated with expanded production to meet increasing demand for animal-source nutrition . In SoutEfforts to meet rising demand for animal-source nutrition in LMICs are driving a shift in animal production from small holder, mixed crop, and livestock operations to increasingly intensive, large-scale, and specialized commercialization ,3. IntenQuantitative volumes of AGP usage from LMICs are not available. However, studies from LMICs indicate substantial nontherapeutic use. In the Mekong Delta of Vietnam, 84% of poultry farms surveyed indicated that antimicrobials were used for prophylactic rather than therapeutic purposes, and nearly a third of overall usage involved antimicrobial classes on the WHO list of highest priority, critically important antimicrobials for human medicine . In the The discovery and commercialization of antimicrobials stands as a defining achievement of 20th century medicine. As usage patterns drive rising rates of antimicrobial resistance (AMR)-linked morbidity and mortality —compoundAntimicrobial usage (AMU) in humans and animals exerts selection pressure potentiating AMR ,5–8; polFacilitates identification of trends and usage profiles,Permits establishment of time-bound consumption targets,Monitors progress toward achieving targets,Guides policy and targeted interventions optimizing antimicrobial use.Monitor consumption trends and benchmarking for optimization of antimicrobial consumption.Comparison of consumption data ultimately disaggregated by species and comparable across human and animal sectors,Antimicrobial consumption data become globally comparable,Contributes to international benchmarking,Creates a globally standardized monitoring architecture, upon which a future international agreement capping consumption could be developed.Comparison of antimicrobial consumption data across countries, species, farms, and with human consumption.Builds a platform for studying the association between antimicrobial consumption and AMR surveillance data.The measurement of AMU in human health and animal health and production settings is a central goal of the Global Action Plan on Antimicrobial Resistance  and the The ecosystem in which veterinary antimicrobials are produced, distributed, and utilized in LMICs is complex and variable. A majority of LMICs currently lack structures capable of quantifying consumption at sufficient resolution to provide usage data by antimicrobial class, animal species, production context, purpose of usage, and route of administration, which are necessary to facilitate effective interventions to optimize use . Of the Patterns of antimicrobial use differ widely across animal production value chains, as evidenced by macrolide, fluoroquinolone, and tetracycline residue detection in animal products marketed for human consumption , but speDespite calls for establishing usage thresholds and targets ,7,17, inOther challenges frequently shared by LMICs  are equaLow rates of AMR awareness and risk perception among farmers and veterinarians ,Inconsistent policies governing antimicrobial use in animal production,Absent AMU regulation and enforcement structures enabling wide accessibility to critically important antimicrobials without prescription,Where a legal veterinarian–client–patient relationship exists governing prescription use of antimicrobials in animals, the link between prescription and sales presents a sales volume profit incentive, andLack of systematic post-market quality surveillance that would enable recall of substandard and counterfeit veterinary antimicrobial products impacting therapeutic efficacy.Multiple methodologies for quantifying usage have been variously employed, hindering data comparability across countries and production sectors. The OIE terrestrial animal health code, the guiding framework for its 180 member nations, outlines the minimum standard as measuring gross usage by weight of active ingredient per year [Through the European Surveillance of Veterinary Antimicrobial Consumption (ESVAC) activity, European Union member states voluntarily report antimicrobial use by food animal–producing species . The ESVvet) and defined course dose in animals (DCDvet) [vet and DCDvet—proprietary ESVAC reporting nomenclature similar to animal defined daily dose (ADDD) and animal defined course dose (ADCD), respectively—are not suited to contexts in which measurement must also capture continuous-use, subtherapeutic dosing, as in LMICs where policies governing AGP use vary significantly.Antimicrobial consumption in food animals can be crudely estimated by quantifying the total weight of the active ingredient adjusted for biomass eligible for treatment, usually expressed as mg/kg population correction unit (PCU), where PCU is defined as total live and slaughtered animal weight per year in a prescribed geographic area. While this measurement of total volume of active ingredient per biomass considers neither the concentration of administered product nor its pharmacokinetics in individual species, it is a readily calculated estimate of usage. ESVAC guidelines use the mg/PCU methodology and introduce both defined daily dose in animals (DDD(DCDvet) , serving(DCDvet) . HoweverAccurate accounting of antimicrobial consumption in food-producing animals of LMICs must capture both therapeutic and nontherapeutic use, including AGPs and mass administration of antimicrobials delivered in medicated, premixed feed. Antimicrobial consumption monitoring is particularly important in aquaculture, in which mass administration of antimicrobials in medicated feed for disease prevention and control is a standard practice .We propose a standardized, internationally-endorsed, phased approach that accounts for the context and currently available structures for collecting AMU data from food-producing animals—both terrestrial and aquatic species—in LMICs. The foundation is built upon the annual measurement of total sales of antimicrobials by class in mg/PCU, which can be derived from the total sales by class for member states reporting to the OIE divided by animal biomass data. Consistency in deriving the numerator—active pharmaceutical ingredient (API)—across multiple formulations of veterinary antimicrobials necessitates that products be managed through a standardized classification scheme, such as the Anatomical Therapeutic Chemical (ATCvet) system established for veterinary medical products . SimilarAs countries establish refined data collection structures , measureIn 2015, Thailand’s International Health Policy Program and One Health partners established a human and animal national Surveillance for Antimicrobial Consumption (Thai SAC) system ,26. ThaiTechnical assistance could be housed within existing or newly established coordinating bodies and should prioritize intraregional LMIC exchanges and solutions relevant to unique regional capacities and contexts. An intergovernmental panel on AMR has been suggested , similarSalmonella enterica Serovar Heidelberg following elimination of third-generation cephalosporins from poultry hatcheries in Quebec, Canada [Documenting usage—particularly for WHO-classified highest priority critically important antimicrobials—permits trends monitoring and identification of animal production sectors in which targeted interventions hold promise of arresting drivers of resistance. Conceivably, a reversion to susceptibility is achievable, as noted in , Canada .Establishing antimicrobial consumption monitoring systems in animal production sectors should be a central, near-term goal of multisectoral national AMR action plans aligned with the WHO Global Action Plan and the focus of advocacy and support from the UN FAO, OIE, and WHO tripartite. Embedding this approach within national AMR action plans promotes monitoring capacities endorsed across ministries, including those vested with budget authority, and costed, resourced, and prioritized for implementation.In parallel, the WHO Joint External Evaluation (JEE) tool, which explicitly considers AMR and is increasingly being utilized by countries as an independent assessment of International Health Regulations (2005) capacity requirements , could dAntimicrobial consumption monitoring systems alone, however, will be insufficient to realize progress in directing prudent AMU in food-producing animals. Concurrently, support for strengthened farm and market chain biosecurity, enhanced livestock vaccination coverage, and improved uptake of good animal husbandry and nutrition practices will be necessary in achieving optimized usage, particularly in transitioning food animal production contexts of LMICs. New production facilities in LMICs should be designed to achieve high standards of husbandry and biosecurity, enabling more rapid phaseout of AGPs. Eliminating AGPs can be achieved at negligible cost to productivity, particularly in the context of such strengthened production systems .The framework presented here should provide LMIC policy makers with high-quality antimicrobial consumption data that can be used to establish usage targets, while building incrementally enhanced monitoring capacities. The analyses derived from this approach could steer targeted policies optimizing antimicrobial consumption and scale back selective pressures currently driving AMR in animal production, with benefits expected to extend broadly across animal and human health.The authors’ views expressed in this publication do not necessarily reflect the views of the United States Agency for International Development or the United States Government.


Solanum lycopersicum) have not been extensively explored. In this study, we characterized the 13 SlGRF genes. In silico analysis of protein motif organization, intron–exon distribution, and phylogenetic classification confirmed the presence of GRF proteins in tomato. The tissue-specific expression analysis revealed that most of the SlGRF genes were preferentially expressed in young and growing tissues such as flower buds and meristems, suggesting that SlGRFs are important during growth and development of these tissues. Some of the SlGRF genes were preferentially expressed in fruits at distinct developmental stages suggesting their involvement in fruit development and the ripening process. The strong and differential expression of different SlGRFs under NaCl, drought, heat, cold, abscisic acid (ABA), and jasmonic acid (JA) treatment, predict possible functions for these genes in stress responses in addition to their growth regulatory functions. Further, differential expression of SlGRF genes upon gibberellic acid (GA3) treatment indicates their probable function in flower development and stress responses through a gibberellic acid (GA)-mediated pathway. The results of this study provide a basis for further functional analysis and characterization of this important gene family in tomato.Growth regulating factors (GRFs) are plant-specific transcription factors that are involved in diverse biological and physiological processes, such as growth, development and stress and hormone responses. However, the roles of GRFs in vegetative and reproductive growth, development and stress responses in tomato ( GRF gene was identified in rice (Oryza sativa) where it mediates gibberellic acid (GA)-induced regulation of stem growth in deepwater rice . EF. EFhttp:GCAG-3’)  was usedeatments .Statistical significance of the differences in relative expression levels of each gene between treatments (control versus stress) and of the differences in expression levels between time points within a treatment was determined with one-way analysis of variance (ANOVA) using the MINITAB statistical software 17 . The mean separation of expression values was analyzed using Tukey’s pairwise comparison test.SlGRF family genes using different bioinformatics approaches and transcript expression analysis. We analyzed their intron–exon organizations, chromosomal distributions, gene structures, evolutionary relationships and expression profiles in different tissues and under different stress conditions to predict their possible biological functions. The SlGRF genes are variably expressed in different tissues and fruits at different developmental stages with particularly high expression in flower buds, and meristems. The increased expression of SlGRF genes in response to abiotic stress and phytohormone treatments implies their function in growth and development of tomato plants under different stress conditions. Together, our results obtained from gene structure, phylogenetic relationships and transcript expression profiles in different tissues and under different stresses facilitate the identification of tomato GRF genes that might play roles in specific developmental processes and/or environmental stress conditions.This study systematically characterized


Negative symptoms (NSs) are more severe in Treatment Resistant Schizophrenia (TRS) than Antipsychotic Responder Schizophrenia (ARS) patients. NSs are predictors of outcomes of neurological soft signs and functional capacity in TRS but not in ARS patients. The scope of this work is to clarify whether NSs effects are integral to or independent from the TRS diagnosis in our sample of patients.70 out of 206 eligible putative TRS and ARS patients were included (enrollment still ongoing). Patients were tested by the Neurological Evaluation Scale (NES); the CGI-S; the PANSS; the Heinrichs’ Quality of Life Scale (QLS); the UCSD Performance-Based Skills Assessment (UPSA); the Personal and Social Performance (PSP) scale and Specific Level of Functioning (SLOF). Patients were subdivided in NSHigh (severe NSs) and NSLow (mild NSs) based on ROC curve-derived cut-off.At the Student’s t test, NSHigh had significantly lower scores than NSLow patients on: Verbal Fluency; QLS score; PSP score; UPSA Financial, Communication, and Family Skills; UPSA total score; all SLOF areas (except Area4). NSHigh patients had significantly higher scores than NSLow patients on CGI-S; PANSS Positive and General Psychopathology Subscale scores; and NES score.Distribution of NS patients was significantly different between TRS/ARS diagnostic groups, as NSHigh patients were significantly more frequent in the TRS group . Notably, mean PANSS Negative Subscale scores were significantly higher in TRS compared to ARS patients .Since multiple variables found to be significantly different in NSHigh vs. NSLow patients were also significantly different between TRS and ARS patients, the question arises whether the significant differences found between diagnostic groups may depend on the higher percentage of patients with more severe NSs in the TRS group. Therefore, a two-way ANOVA was carried out with dichotomous NS and Diagnosis variables as the independent variables. Outcomes on multiple clinical variables were significantly different among groups. A NS*Diagnosis interaction effect was found for NES score , Passive Social Withdrawal (N4), and Difficulty in Abstract Thinking (N5).These data suggest that NSs are both independent determinants and moderators of TRS/ARS diagnosis effect on multiple psychopathology, cognitive, and psychosocial factors. More impaired functions attributed to non-response to antipsychotics may depend on more severe NSs. However, only a subset of NSs appears to exert this action, possibly related to the multidimensional construct of these symptoms.


Malignant hypertension may be the first manifestation of systemic hypertension. We report a clinical case of a Caucasian 41-year-old man with no previous history of blood hypertension seen at casualty because of blurred vision. Fundus examination disclosed optic disk swelling, retinal hemorrhages and infarcts. The blood pressure was 220/130 mmHg. After the appropriate management of hypertension, optic disk and retinal edema resolved, leaving minor changes as mild optic disk pallor and hard exudates.


Atherosclerotic stenosis of the brachiocephalic artery sometimes occurs in patients with coronary artery disease, and can cause stroke during the perioperative period of coronary artery bypass grafting.We describe the case of a 77-year old male with severe stenosis of the brachiocephalic artery and severe coronary artery disease. He successfully underwent aorto-right subclavian artery bypass that was performed concomitantly with off-pump coronary artery bypass.Concomitant aorto-subclavian artery bypass with off-pump coronary artery bypass grafting is a therapeutic option that minimizes the risk of perioperative stroke in patients with brachiocephalic artery stenosis and coronary artery disease. Atherosclerotic stenosis of the brachiocephalic artery can be associated with coronary artery disease . StenosiA 77-year old male was referred to our hospital for known coronary artery disease and a history of antero-septal myocardial infarction and syncope. Coronary angiography showed severe triple vessel disease. Computed tomography revealed severe diffuse calcification from the aortic arch to the abdominal aorta. In particular, just proximal brachiocephalic artery was shown to be circumferentially calcified and 90% stenosed Fig. . ProximaA median sternotomy was performed, and the bilateral internal thoracic arteries were dissected. At the same time, a saphenous vein graft was dissected. A direct ultrasonographic examination revealed severe atheromatous plaques inside the brachiocephalic artery; hence, we chose the right subclavian artery as the distal anastomosis site for the prosthetic bypass conduit. An 8-mm polytetrafluoroethylene graft was anastomosed to the right subclavian artery, and then the proximal end of this graft was anastomosed to the aorta with a side-biting clamp. Off-pump CABG was then performed using a method described previously . The lefPostoperatively, no neurological deficit was observed. The blood pressures of the bilateral upper extremities were identical. Computed tomography demonstrated the patency of the coronary bypass grafts and the aorto-right subclavian artery bypass graft Fig. . PostopeAtherosclerotic stenosis of the subclavian artery or brachiocephalic trunk is sometimes associated with coronary artery disease . Many paPreoperative computed tomography revealed severe calcification just proximal brachiocephalic artery, and carotid ultra sound examination revealed subclavian steal phenomenon at right vertebral artery. But computed tomography revealed no significant stenosis from end of the brachiocephalic trunk to origin of right carotid artery and subclavian artery, and so we expected that the aorto-subclavian bypass provided a physiological brain perfusion and reduced the risk of stroke.Takach et al.  reportedConcomitant aorto-subclavian artery bypass with off-pump CABG is a therapeutic option that minimizes the risk of perioperative stroke in patients with brachiocephalic artery stenosis and coronary artery disease.


Cyclosporin-A (CsA) and tacrolimus (FK-506) are immunomodulating agents used to prevent rejection in organ transplantation. They are both associated with several side effects, including nephrotoxicity and severe hypertension due to vascular injury, which often appears as a microvascular occlusive disorder . We report the first case of a microvascular occlusive disorder with the features of TMA in the small bowel of an orthotopic liver transplant (OLT) patient after immunosuppressive therapy with CsA and FK506. The patient presented with severe recurrent abdominal colics and distal subocclusion, requiring aggressive surgical treatment. Histological and ultrastructural analysis of the resected specimen disclosed intestinal TMA. Although rare, such a complication should be considered in the differential diagnosis of abdominal colics in patients undergoing immunosuppressant therapy after OLT.


Biotic and abiotic factors are increasingly acknowledged to synergistically shape broad-scale species distributions. However, the relative importance of biotic and abiotic factors in predicting species distributions is unclear. In particular, biotic factors, such as predation and vegetation, including those resulting from anthropogenic land-use change, are underrepresented in species distribution modeling, but could improve model predictions. Using generalized linear models and model selection techniques, we used 129 estimates of population density of wild pigs (Sus scrofa) from 5 continents to evaluate the relative importance, magnitude, and direction of biotic and abiotic factors in predicting population density of an invasive large mammal with a global distribution. Incorporating diverse biotic factors, including agriculture, vegetation cover, and large carnivore richness, into species distribution modeling substantially improved model fit and predictions. Abiotic factors, including precipitation and potential evapotranspiration, were also important predictors. The predictive map of population density revealed wide-ranging potential for an invasive large mammal to expand its distribution globally. This information can be used to proactively create conservation/management plans to control future invasions. Our study demonstrates that the ongoing paradigm shift, which recognizes that both biotic and abiotic factors shape species distributions across broad scales, can be advanced by incorporating diverse biotic factors. Predicting and mapping species distributions, including geographic range and variability in abundance, is fundamental to the conservation and management of biodiversity and landscapes23There is an ongoing paradigm shift in understanding how biotic and abiotic factors shape species distributions. Until recently, it was widely accepted that abiotic factors, such as temperature and precipitation, played the primary role in shaping distributions of species and biodiversity at broad scales  and that biotic factors were most important at fine scales 9101213141515197In addition to species interactions, biotic factors related to vegetation can influence species distributions and abundance at broad scales. In particular, anthropogenic land-use change is rarely considered when evaluating species distributions at broad scales; however, given the human footprint globally23252613Invasive species are a primary driver of widespread and severe negative impacts to ecosystems, agriculture, and humans across local to global scalesMus musculus) and brown rat (Rattus norvegicus), wild pigs  have one of the widest geographic distributions of any mammal; further, it exhibits the widest geographic range of any large mammal353940Few species exhibit a global distribution that extends across Europe, Asia, Africa, North and South America, Australia, and oceanic Islands. Besides naturalized animals, such as the house mouse  evaluate how biotic  and abiotic  factors  shape po2), which resulted in 129 estimates of density across their global distribution used in our analyses  had > 1,000 times more support as the best approximating model than the top model considering only abiotic factors   . The var× 10−17) . Density× 10−17) ; percentUsing the full model-averaged results of parameter estimates, we created a predictive map of global wild pig population density . Wild piPopulation density of an invasive large mammal was strongly influenced by both biotic and abiotic factors across its global distribution. Consistent with the prediction that abiotic factors drive broad-scale patterns of species distribution, potential evapotranspiration (PET) and precipitation variables were important predictors of population density on a global scale. In addition, contributing to growing evidence that biotic factors are also important determinants of broad-scale patterns of species distributions, both biotic interactions and vegetation played important roles in predicting the distribution of wild pig populations globally. Further, land-use change mediated by human activities strongly predicted the broad-scale distribution of an invasive large mammal. Consistent with previous studies evaluating how population density of ungulates varied across broad scales, both bottom-up (resource-related) and top-down (predation) factors influenced the distribution of wild pig populations1942Using both biotic and abiotic factors to evaluate broad-scale species distributions can create more realistic maps of range and density with better predictive ability16Abiotic factors, such as temperature and precipitation, are consistently found to be primary determinants of species distributions at broad scalesBiotic factors were among the most supported variables predicting population density across a global scale. Our results indicated that the presence of large carnivores can influence wild pig population density. Large carnivore richness was strongly supported in our models and exhibited a negative relationship with wild pig density; as the number of large carnivore species increased, wild pig density decreased, which is consistent with studies in Eurasia and Australia4247495052531718Although biotic interactions between animals are the primary biotic factors evaluated in species distribution models at broad scales, the role of plant communities has received less consideration. In particular, anthropogenic land-use change increasingly influences vegetation communities across continents and warrants a better understanding for how human activities are shaping broad-scale distributions of plant and animal populations22235758Forest is considered a key habitat type preferred by wild pigs59In some systems, abiotic factors can be stronger predictors of species distributions, than biotic factors, because of high correlations between these two factorsAdditional biotic factors that can influences species distributions on a broad scale, particularly invasive species, include the role of humans in distributing the founding individuals of new populations. For example, invasive wild pig populations have arisen across several continents recently through human activities. Illegal translocations by humans for hunting purposes can facilitate the long-distance expansion of wild pig populations into new areas646539676860Population density, compared to presence-absence occurrence, can provide more informative conclusions of species distributions in relation to biotic and abiotic factors77374Predicting species distributions provides critical information to the management and conservation of biodiversity, especially for controlling invasive species. Without intensive management actions, our study predicts that there is strong potential for wild pigs to expand their geographic range and further invade expansive areas of North America, South America, Africa, and Australia. Although wild pigs currently occupy broad regions of predicted habitat in their non-native range, many regions of predicted habitat are currently unoccupied and may be at high risk for future invasion. These areas might warrant increased surveillance by local, state, and federal agencies to counter the establishment of populations. Although attention in unoccupied areas that are predicted to support high densities of wild pigs might warrant priority for countering population introductions, wild pigs can persist in relatively low quality habitat  and these areas also warrant attention to halt invasions. Given the potential for wild pig populations to rapidly expand once establishedet al.76To evaluate the population density  of wild pigs throughout their global distribution, we compiled density estimates from the literature throughout its native and non-native ranges across each continent and island for which data were available . PreviouModels evaluating and predicting species distributions can be improved by including areas of absence  or zero density to sample the full range of available landscape conditions73We considered a suite of biotic and abiotic landscape variables, which were divided into vegetation, predation, and climate factors  that we The biotic factors that we evaluated included agriculture, broadleaf forest, enhanced vegetation index (EVI), forest canopy cover, difference in the proportion between forest and agriculture (to characterize landscape heterogeneity), normalized difference vegetation index (NDVI), large carnivore richness, and unvegetated area . We expeThe abiotic factors that we evaluated included two measures of ecological energy regimes, actual evapotranspiration  and potential evapotranspiration , and calculated variable importance values93942 resolution). This map displays the maximal potential density of wild pigs in relation to the biotic and abiotic factors used in our modeling and reflects predicted densities that would be achieved if wild pigs had access to all landscapes, their movements were unrestricted, and management activities did not suppress populations. We validated our model using mean squared prediction error (MSPE)We used multiple linear regression to evaluate how population density was influenced by our final suite of biotic and abiotic factors . The disHow to cite this article: Lewis, J. S. et al. Biotic and abiotic factors predicting the global distribution and population density of an invasive large mammal. Sci. Rep.7, 44152; doi: 10.1038/srep44152 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


In our recent updated studies, we found a significant association between periprocedural blood transfusion and acute kidney injury (AKI) following transcatheter aortic valve replacement (TAVR) with an overall 1.95-fold increased the risk of AKI. We also demonstrated that a transapical approach was significantly associated with increased AKI risk compared with a transfemoral approach. Nevertheless, the TAVR approach did not affect severe renal outcomes or long-term renal function. In addition, our meta-analysis demonstrated no significant association between contrast media volume and risk of AKI after TAVR. Thus, the dose of contrast media likely does not play a significant role in the pathogenesis of TAVR-related AKI. Growing knowledge of these risk factors of TAVR on kidney function will help improve preventive measures to improve patients’ outcomes.We agree with Onuigbo and Agbasi that previous studies have attempted to identify effective interventions to prevent postoperative acute kidney injury (AKI) events including cardiac surgery-associated AKI and most of them failed to be successful ,2. In thAll authors have contributed equally to the preparation of the manuscript.The authors declare no conflict of interest.Ethical issues  have been completely observed by the authors.None.


Oncorhynchus spp.) carcasses might yield suitable DNA quality for noninvasive monitoring of brown bears (Ursus arctos). We compared the efficiency of monitoring brown bear populations using both fecal DNA and salivary eDNA collected from partially-consumed salmon carcasses in Southeast Alaska. We swabbed a range of tissue types from 156 partially-consumed salmon carcasses from a midseason run of lakeshore-spawning sockeye (O. nerka) and a late season run of stream-spawning chum (O. keta) salmon in 2014. We also swabbed a total of 272 scats from the same locations. Saliva swabs collected from the braincases of salmon had the best amplification rate, followed by swabs taken from individual bite holes. Saliva collected from salmon carcasses identified unique individuals more quickly and required much less labor to locate than scat samples. Salmon carcass swabbing is a promising method to aid in efficient and affordable monitoring of bear populations, and suggests that the swabbing of food remains or consumed baits from other animals may be an additional cost-effective and valuable tool in the study of the ecology and population biology of many elusive and/or wide-ranging species.Noninvasive genetic sampling is an important tool in wildlife ecology and management, typically relying on hair snaring or scat sampling techniques, but hair snaring is labor and cost intensive, and scats yield relatively low quality DNA. New approaches utilizing environmental DNA (eDNA) may provide supplementary, cost-effective tools for noninvasive genetic sampling. We tested whether eDNA from residual saliva on partially-consumed Pacific salmon ( Environmental DNA, or eDNA, has proven to be a comprehensive, noninvasive means of monitoring biodiversity, providing rapid, cost-effective, and efficient insights on species’ distribution and abundance ,2. As a et al. , marten [Martes americana], etc.), particularly if the microsatellites used are shared among species. For example, the sex marker we used, SRY, amplifies in multiple mammalian species. Where contamination is possible, researchers should consider utilizing species-specific markers. Studies utilizing markers that overlap among species should ensure that all PCR replicates give consistent results. These considerations aside, saliva is a promising new source for bear DNA in salmon ecosystems.Additionally, because of the nature of bear feeding behavior, it is possible for carcasses to become contaminated with DNA from a second individual bear if scavenging occurs following the initial consumption event. In populations with high allelic diversity and heterozygosity, however, mixed samples should be detectable . ContamiWe found residual saliva collected from partially-consumed salmon carcasses to be a successful, labor- and cost-effective way to sample brown bear populations. As far as we are aware, this study is the first to use noninvasively collected saliva to estimate the population density of any species of wildlife. As we focused primarily on the methodology of saliva sampling in this study, we recommend that researchers looking to implement this technique as a method to monitor bear populations take into account potential capture heterogeneity and violation of assumptions for mark-recapture models that may arise due to differing levels of fishing success or consumption behaviors among individual bears.Although salmon carcasses provide a readily available source of saliva to aid in the monitoring of bear populations, baited saliva sampling should be easily generalizable to systematic surveys of carnivores if appropriate baits can be identified and tested . As an eS1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S1 FigNumber of samples that successfully genotyped  versus number of unique individuals identified  across all samples collected, with trendlines.(TIF)Click here for additional data file.S1 File(DOCX)Click here for additional data file.


Wolbachia (Alphaproteobacteria) and Spiroplasma (Mollicutes), are discussed. Although several general patterns of genome reduction associated with the adoption of symbiotic relationships could be identified, extensive variation was found among these facultative symbionts. These findings are incorporated into the established conceptual frameworks to develop a more detailed evolutionary model for the discussion of possible trajectories. In summary, transitions from facultative to obligate symbiosis do not appear to be a universal one-way street; switches between hosts and lifestyles  occur frequently and could be facilitated by horizontal gene transfer.Symbiosis between organisms is an important driving force in evolution. Among the diverse relationships described, extensive progress has been made in insect–bacteria symbiosis, which improved our understanding of the genome evolution in host-associated bacteria. Particularly, investigations on several obligate mutualists have pushed the limits of what we know about the minimal genomes for sustaining cellular life. To bridge the gap between those obligate symbionts with extremely reduced genomes and their non-host-restricted ancestors, this review focuses on the recent progress in genome characterization of facultative insect symbionts. Notable cases representing various types and stages of host associations, including those from multiple genera in the family Enterobacteriaceae (class Gammaproteobacteria),  This review synthesizes the recent progress in genome characterization of insect-symbiotic bacteria, the emphases include (i) patterns of genome organization, (ii) evolutionary models and trajectories, and (iii) comparisons between facultative and obligate symbionts. None declared.


Morphological responses of nonmammalian herbivores to external ecological drivers havenot been quantified over extended timescales. Herbivorous nonavian dinosaurs are an idealgroup to test for such responses, because they dominated terrestrial ecosystems for morethan 155 Myr and included the largest herbivores that ever existed. The radiation ofdinosaurs was punctuated by several ecologically important events, including extinctionsat the Triassic/Jurassic (Tr/J) and Jurassic/Cretaceous (J/K) boundaries, the decline ofcycadophytes, and the origin of angiosperms, all of which may have had profoundconsequences for herbivore communities. Here we present the first analysis ofmorphological and biomechanical disparity for sauropodomorph and ornithischian dinosaursin order to investigate patterns of jaw shape and function through time. We find thatmorphological and biomechanical mandibular disparity are decoupled: mandibular shapedisparity follows taxonomic diversity, with a steady increase through the Mesozoic. Bycontrast, biomechanical disparity builds to a peak in the Late Jurassic that correspondsto increased functional variation among sauropods. The reduction in biomechanicaldisparity following this peak coincides with the J/K extinction, the associated loss ofsauropod and stegosaur diversity, and the decline of cycadophytes. We find no specificcorrespondence between biomechanical disparity and the proliferation of angiosperms.Continual ecological and functional replacement of pre-existing taxa accounts fordisparity patterns through much of the Cretaceous, with the exception of several uniquegroups, such as psittacosaurids that are never replaced in their biomechanical ormorphological profiles. Sauropodomorph and ornithischian dinosaurs were the foremost herbivorous terrestrialvertebrates of the Mesozoic Era in terms of species richness, abundance, and functionaldiversity  in herbivorous nonavian dinosaursthrough time. This approach complements previous attempts to examine these questions thoughspatiotemporal comparisons of species-richness patterns and provides the only rigorousbiomechanically and functionally based analysis of these issues attempted to date. Wehypothesize that ornithischians and sauropodomorphs will show distinct morphologies andbiomechanical profiles . Wealso hypothesize that the shift in plant community structure after the J/K boundary willtrigger a corresponding shift in dinosaurian jaw biomechanical profiles, due to thediffering physiognomies, digestibility, and mechanical properties of the varied potentialfood plant clades that were ecologically important at different times throughout theMesozoic Bakker . We use Data for 2D landmark and biomechanical trait analyses were compiled from 167 sauropodomorphand ornithischian dinosaur taxa . Herbivorousnonavian theropods were excluded from this data set, as complete mandibular material forthese animals is rare. A mandibular biomechanical profile represents a good proxy forcharacterizing the feeding system, as the mandible is primarily adapted for feeding, whereasthe cranium has multiple functional roles, some of which are unrelated to feeding, such ashousing the brain and sensory organs  in Past, Version 3 , with the exception of Stegosaurus (twospecies). Regions of overlap are occupied by a wide range of both basal and derivedsauropodomorphs; these include: Plateosaurus gracilis,Lamplughsaura, mamenchisaurids, brachiosaurids, and two SouthAmerican titanosaurids (Antarctosaurus andBonitasaura). Sauropodomorphs occupy morphospace exclusively in the−PC1 region: this region is characterized by dorsoventrally narrow jaws and the lack ofa prominent coronoid process. Noneusauropod sauropodomorphs , for the most part,account for sauropodomorph occupation of morphospace in +PC2: this region is typified byvery narrow anterior jaws. Macronarian and diplodocoid taxa (includingDiplodocus and Tapuiasaurus) primarily occupy −PC2regions of morphospace . Jaws in this region exhibit a greater gap betweenlandmarks 1 and 2 than in sauropodomorph morphospace (due to the presence of thepredentary in iguanodontians). Disparate groups of nonthyreophoran ornithischians expandmorphospace occupation into +PC1 and +PC2 (hadrosaurids) and −PC2 regions(leptoceratopsids and psittacosaurids). +PC1 and +PC2 regions typically contain jawswith prominent coronoid processes and downwardly deflected predentaries; −PC2 regionscontain robust, dorsoventrally broad jaws. Nonceratopsid marginocephalian jawmorphologies, such as those of psittacosaurids and leptoceratopsids, contribute stronglyto the expansion of ornithischian shape morphospace, predominantly into +PC1/−PC2. Taxaare absent in a region of morphospace around +0.05 PC1/−0.075 PC2.Our results demonstrate that sauropodomorph and ornithischian jaws occupy significantlydifferent regions of morphological morphospace p<0.01; ; Table2phospace . The cenp<0.01; Pisanosaurus, heterodontosaurids) and basal members of Thyreophora, Marginocephalia (Yinlong),and Ornithopoda .Sauropodomorphs occupy regions of +PCo1. Noneusauropod sauropodomorphs  predominate in+PCo1/−PCo2. This region is characterized by jaws with a high mechanical advantage and alarge adductor muscle attachment area. Diplodocids, nonneosauropods, andnontitanosaurian macronarians  stretch sauropodomorph occupation into +PCo2. Jaws inthis region also display high mechanical advantages, coupled with high aspect ratios.Many iguanodontian, ceratopsid, and psittacosaurid jaw profiles occupy similar regionsof +PCo2 biomechanical morphospace (Montanoceratops). This region of functional space is characterized bydeep jaws with short adductor muscle attachment and a high posterior mechanicaladvantage. Expansion into −PCo2 is accounted for by deep-jawed ankylosaurs, with low tooth:jawdepth ratios and high relative dental length  share areas ofbiomechanical morphospace with iguanodontians but occupy very different regions of shapespace  that plots between nonhadrosaurid ornithopods andankylosaurians  extend biomechanical morphospace occupation into theregion of morphospace characterized by deep mandibles with short adductor muscleattachment and high posterior mechanical advantages . By contrast,biomechanical disparity undulates through the Mesozoic . Thereare no significant differences in disparity between successive time bins for eitherbiomechanical or morphological disparity curves (at p=0.05) and nomarginal-likelihood values exceed the threshold value of 8. There are a few instanceswhere disparity diverges markedly from sample size, suggesting that a trend, albeitnonsignificant, might be observed. For example, morphological disparity rises in theEarly Cretaceous, immediately after the J/K extinction, and in the early LateCretaceous, while sample size drops. Likewise, biomechanical disparity drops in theMiddle Jurassic while sample size rises slightly. Conversely, in the latest Cretaceous,sample size rises sharply while biomechanical disparity drops very slightly.Morphological (shape) and biomechanical disparity measures are decoupled through theMesozoic . MorpholMesozoic , a decoun=5). The lack of manydinosaur-bearing formations between the Berriasian and Albian may partially account forthe low species richness observed in this interval, although it could also be attributedto the J/K extinction event , despite the former existing around 10Myr earlier: this pattern supports theresults of another recent quantitative craniodental study . Perhapssurprisingly, we find minimal convergent occupation in biomechanical morphospace betweentitanosaurids  and diplodocids   and titanosaurids with longer snouts and pencil-liketeeth (such as Antarctosaurus), and diplodocids are outliers in thisbiomechanical morphospace. This pattern supports quantitative work on sauropodomorphcranial morphology related to feeding, with similar placement of the same taxa incranial  and Manidens(Heterodontosauridae) share occupation of Late Jurassic sauropodomorph biomechanicalmorphospace . This suggests that mandibles withsimilar gross morphology were biomechanically and functionally differentiated by thistime. In general, sauropodomorphs and heterodontosaurids occupy similar regions of bothshape-based and biomechanical morphospace and do not extend their occupation ofmorphospace beyond regions already occupied by the end of the Early Jurassic (Camptosaurus), which is also reflected in the morphological disparitycurve  occupy novel regions of morphological andbiomechanical morphospace: these taxa share regions of biomechanical morphospace withhadrosauroids until the disappearance of basal marginocephalians prior to the last 20Myr of the Mesozoic (Triceratops) and leptoceratopsids . The biomechanical profiles of ceratopsids show nooverlap with those of hadrosaurids. This supports the conclusions of Mallon and Anderson leads to an increase in biomechanical disparity levels fromthe early Late Cretaceous. Marginocephalian, ornithopod, and thyreophoran biomechanicalmorphospace occupation in the latest Cretaceous suggests that these groups, whilevarying from each other in mandibular shape, also share a variety of functional andbiomechanical traits relating to feeding. Late Cretaceous hadrosaurids and ankylosauridsfilled the biomechanical roles vacated by Early Cretaceous nonhadrosaurid iguanodontiansand nodosaurids, respectively. Individual occupation of morphospace by each taxon can beviewed in Supplementary Figures 2–6.Early Cretaceous marginocephalians  . The exclusion of the Morrisontaxa removes the Late Jurassic peak in biomechanical disparity .A similar jackknifing of the taxa from the Dashanpu Formation (including the “Upper andLower Shaximiao” formations) yielded a trough in disparity in the Middle Jurassic butretained a strong peak in the latest Jurassic . These resultssuggest that the data may be sensitive to the inclusion or exclusion of particularlyrich fossil-bearing sites. In addition, the lack of available jaw material from Northand South American titanosaurs seriously underrepresents sauropodomorph diversity in theCretaceous. The addition of titanosaurid taxa to the analysis may increase both thedisparity and overall morphospace occupation of sauropodomorphs, although the titanosaurjaws sampled in this study already account for a broad range of morphologies.When disparity tracks sample diversity closely, as it does in this study forshape-based disparity, sampling bias cannot be ruled out. Morphological disparity inthis study partly tracks jaw sample size, suggesting a potential bias in the data setfor some features of the disparity curve  when compared with mid-browsing taxa , and almost equal in disparity to very low-browsingsauropodomorphs . This pattern contrasts with sample diversity, with thelowest sample size found in the high-browsing feeding envelope (n=6). Unfortunately, low sample sizes within each feeding levelprevent any significant differences or definitive conclusions to be made. However, thispattern remains intriguing and the addition of more mandibular remains from high- andmid-browsing taxa to our sample (as and when they are discovered) would complement thisstudy. This is an avenue of study that requires more investigation in the future toenable deeper insights into niche partitioning between sauropod groups based on maximumbrowse height.Supplementary analyses of biomechanical and shape-based disparity withinsauropodomorphs in relation to maximum feeding height show higher levels of disparity inhigh-browsing sauropods .The dip in biomechanical disparity after the J/K recovered by our analyses may,therefore, be an artefact due to either geological biases or uneven collection effort,underrepresenting the true diversity of jaw biomechanical profiles at this time. Due tothe lack of complete mandibles from rebbachisaurids, dicraeosaurids, and other clades,it is possible that the latest Jurassic and earliest Cretaceous disparity levelsreported herein are currently undersampling the total diversity of mandible morphologyand potential function. Such exclusions cannot be corrected for by our analyses andrepresent a limitation of the fossil material currently available.Udanoceratops), a group that is also present in North America. LateCretaceous hadrosaurids and ankylosaurids filled the biomechanical roles vacated by EarlyCretaceous nonhadrosaurid iguanodontians and nodosaurids respectively. Our results implythat, after the establishment of peak overall biomechanical variation in the latestJurassic, only marginocephalians demonstrated widespread variation in biomechanical profilesover time, triggered by the isolated adaptive radiations of psittacosaurids andleptoceratopsians. The remainder of Cretaceous herbivorous dinosaurs underwent progressiveniche replacement, with successive replacement by related taxa with comparable biomechanicalprofiles.For the first time, we have quantified the morphological and biomechanical variation ofornithischian and sauropodomorph jaws throughout the Mesozoic and examined how diversityrelated to external extrinsic drivers such as extinction events and the rise of angiosperms.We find that herbivorous dinosaur clades have jaws that occupy different regions ofmorphospace throughout the Mesozoic. Furthermore, sauropodomorphs and ornithischians havejaws that also function in broadly different ways, yet there is some potentially convergentoverlap in biomechanical function between different ornithischian clades in the Cretaceous.Basal members of each clade tend to be more similar in form and function to each other,while derived taxa are more functionally and morphologically divergent. Herbivorous dinosaurjaws maintained a numerically steady diversity of biomechanical traits, with a peak observedin the Late Jurassic triggered by the diversification of high-browsing sauropods. This isconsistent with a rapid evolutionary radiation in biomechanical diversity among herbivorousdinosaurs followed by a plateau. The Tr/J extinction had no overall effect on biomechanicalvariation among herbivorous dinosaurs, despite fundamental changes in floral and faunalcomposition across the boundary. This consistency suggests that Early Jurassic dinosaursfilled the functional feeding niches vacated by the extinction of Late Triassic taxa.Similar successive replacement patterns are also seen in Devonian gnathostomes and Devonianto mid-Pennsylvanian tetrapodomorphs (Anderson et al.