Efficient delivery of anticancer drugs using functionalized-Ag-decorated Fe3O4@SiO2 nanocarrier with folic acid and ß-cyclodextrin.

Nanocarrier surface functionalization has been widely regarded as a promising approach for achieving precise and targeted drug delivery systems. In this work, the fabrication of functionalized-Ag-decorated Fe3O4@SiO2 (Fe3O4@SiO2-Ag) nanocarriers with folic acid (FA) and ß-cyclodextrin (BCD) exhibit a remarkable capacity for delivering two types of anticancer drugs, i.e., doxorubicin (DOX) and epirubicin (EPI), into cancer cells. The effective functionalization of Fe3O4@SiO2-Ag nanoparticles has been achieved through the use of cysteine (Cys) as an anchor for attaching FA and BCD via EDC-NHS coupling and Steglich esterification methods, respectively. The findings indicate that surface functionalization had no significant impact on the physicochemical characteristics of the nanoparticles. However, it notably affected DOX and EPI loading and release efficiency. The electrostatic conjugation of DOX/EPI onto the surface of Fe3O4@SiO2-Ag/Cys/FA and Fe3O4@SiO2-Ag/Cys/BCD exhibited maximum loading efficiency of 50-60% at concentration ratio of DOX/EPI to nanoparticles of 1:14. These nanocarriers also achieved an 40-47% DOX/EPI release over 36 days. Furthermore, the drug-loaded functionalized-nanocarrier showed cytotoxic effects on SK-MEL-2 cells, as demonstrated by an in vitro MTT assay. This suggests that the as-prepared functionalized-nanoparticles have promise as a carrier for the efficient anticancer drugs.

Simultaneous ocean acidification and warming do not alter the lipid-associated biochemistry but induce enzyme activities in an asterinid starfish.

Ocean acidification and warming affect marine ecosystems from the molecular scale in organismal physiology to broad alterations of ecosystem functions. However, knowledge of their combined effects on tropical-subtropical intertidal species remains limited. Pushing the environmental range of marine species away from the optimum initiates stress impacting biochemical metabolic characteristics, with consequences on lipid-associated and enzyme biochemistry. This study investigates lipid-associated fatty acids (FAs) and enzyme activities involved in biomineralization of the tropical-subtropical starfish Aquilonastra yairi in response to projected near-future global change. The starfish were acclimatized to two temperature levels (27 °C, 32 °C) crossed with three pCO2 concentrations (455 µatm, 1052 µatm, 2066 µatm). Total lipid (ΣLC) and FAs composition were unaffected by combined elevated temperature and pCO2, but at elevated temperature, there was an increase in ΣLC, SFAs (saturated FAs) and PUFAs (polyunsaturated FAs), and a decrease in MUFAs (monounsaturated FAs). However, temperature was the sole factor to significantly alter SFAs composition. Positive parabolic responses of Ca-ATPase and Mg-ATPase enzyme activities were detected at 27 °C with elevated pCO2, while stable enzyme activities were observed at 32 °C with elevated pCO2. Our results indicate that the lipid-associated biochemistry of A. yairi is resilient and capable of coping with near-future ocean acidification and warming. However, the calcification-related enzymes Ca-ATPase and Mg-ATPase activity appear to be more sensitive to pCO2/pH changes, leading to vulnerability concerning the skeletal structure.

Effect of Aspect Ratio of a Gold-Nanorod-Modified Screen-Printed Carbon Electrode for Carbaryl Detection in Three Different Samples of Vegetables.

In this study, three different sizes of gold nanorods (AuNRs) were synthesized using the seed-growth method by adding various volumes of AgNO3 as 400, 600, and 800 µL into the growth solution of gold nanoparticles. Three different sizes of AuNRs were then characterized using UV-vis spectroscopy, high-resolution transmission electron microscopy (HRTEM), selected area electron diffraction (SAED) patterns, and atomic force microscopy (AFM) to investigate the surface morphology, topography, and aspect ratios of each synthesized AuNR. The aspect ratios from the histogram of size distributions of three AuNRs as 2.21, 2.53, and 2.85 can be calculated corresponding to the addition of AgNO3 volumes of 400, 600, and 800 µL. Moreover, each AuNR in three different aspect ratios was drop-cast onto the surface of a commercial screen-printed carbon electrode (SPCE) to obtain three different SPCE-modified AuNRs (SPCE-A400, SPCE-A600, and SPCE-A800, respectively). All SPCE-modified AuNRs were then evaluated for their electrochemical behavior using cyclic voltammetry and electrochemical impedance spectroscopy (EIS) techniques and the highest electrochemical performance was shown as the order of magnitude of SPCE-A400 > SPCE-A600/SPCE-A800. The reason for the highest electrocatalytic activity of SPCE-A400 might be due to the smallest particle size and uniform distribution of AuNRs ∼ 2.2, which enhanced the charge transfer, thus providing the highest electroactive surface area (0.6685 cm2) compared to other electrodes. These results also confirm that the sensing mechanism for all SPCE-modified AuNRs is controlled by diffusion phenomena. In addition, the optimum pH was obtained as 4 for carbaryl detection for all SPCE-modified AuNRs with the highest current shown by SPCE-A400. Furthermore, SPCE-A400 has the highest fundamental parameters (surface coverage, catalytic rate constant, electron transfer rate constant, and adsorption capacity) for carbaryl detection, which were investigated using cyclic voltammetry and chronoamperometric techniques. The electroanalytical performances of all SPCE-modified AuNRs for carbaryl detection were also investigated with SPCE-A400 displaying the best performance among other electrodes in terms of its linearity (0.2-100 µM), limit of detection (LOD) ∼ 0.07 µM, and limit of quantification (LOQ) ∼ 0.2 µM. All SPCE-modified AuNRs were also subsequently evaluated for their stability, reproducibility, and selectivity in the presence of interfering species such as NaNO2, NH4NO3, Zn(CH3CO2)2, FeSO4, diazinon, and glucose and show reliable results as depicted from %RSD values less than 3%. At last, all SPCE-modified AuNRs have been employed for carbaryl detection using a standard addition technique in three different samples of vegetables (cabbage, cucumber, and Chinese cabbage) with its results (%recovery ≈ 100%) within the acceptable analytical range. In conclusion, this work demonstrates the great potential of a disposable device based on an AuNR-modified SPCE for rapid detection and high sensitivity in monitoring the concentration of carbaryl as a residual pesticide in vegetable samples.

Antioxidant Activity and Metabolite Profiling of Xylocarpus granatum Extracts Using Gas Chromatography-Mass Spectrometry.

The potential application of Xylocarpus granatum, a mangrove species, as traditional medicine has been widely linked to its high secondary metabolite and antioxidant contents. However, few studies have been reported to identify and classify active metabolites responsible for such excellent biological activities. Therefore, the aim of this work was to determine the antioxidant activity, identify the metabolite profiles, and predict the metabolites acting as antioxidants in X. granatum extract using a gas chromatography-mass spectrometry (GC-MS)-based metabolomics approach. The seeds, stems, fruit peel, pulp, leaves, and twigs of X. granatum were macerated with ethanol. Each extract was analyzed with GC-MS, and the data were processed using mass spectrometry data-independent analysis (MS-DIAL) software to identify the metabolites. The IC50 value of plant parts of X. granatum ranged from 7.73 to 295 ppm. A total of 153 metabolites were identified and confirmed in the X. granatum extracts. Among the identified metabolites, epicatechin and epigallocatechin were the two most abundant in the stem extracts and are expected to have the greatest potential as antioxidants. Principal component analysis (PCA) succeeded in grouping all parts of the plant into three groups based on the composition of the metabolites: group 1 (stems, fruit peel, and twigs), group 2 (seeds and pulp), and group 3 (leaves).

Colloidal Stability of CA, SDS and PVA Coated Iron Oxide Nanoparticles (IONPs): Effect of Molar Ratio and Salinity.

Iron Oxide Nanoparticles (IONPs) have received unprecedented interest in various applications. The main challenges in IONPs are fluid stability due to agglomeration in a saline condition. This paper aims to investigate the colloidal stability of citric acid (CA), sodium dodecyl sulphate (SDS) and polyvinyl alcohol (PVA) under various molar ratios and levels of salinity. Firstly, the IONPs were synthesized using a facile co-precipitation approach. Secondly, the IONPs were coated using a simple dip-coating method by varying the molar ratio of CA, SDS and PVA. Next, the coated IONPs were characterized by using an X-ray Diffractometer (XRD), Fourier transform infrared spectroscopy (FTIR), and a Field Emission Scanning Electron Microscope (FESEM) for the morphological and crystallographic study of coated IONPs. Finally, the coated IONPs were characterized for their zeta potential value and hydrodynamic size using a Zetasizer and their turbidity was measured using a turbidity meter. It was found that at a low salinity level, 0.07 M of CA-IONPs, a high zeta potential value, a smaller hydrodynamic size, and a high turbidity value of -40.9 mV, 192 nm and 159 NTU were observed, respectively. At a high salinity level, 1.0 M SDS-IONPs recorded a high zeta potential value of 23.63 mV, which corresponds to a smaller hydrodynamic size (3955 nm) and high turbidity result (639 NTU). These findings are beneficial for delivering cutting-edge knowledge, especially in enhanced oil recovery (EOR) applications.

Selective non-enzymatic uric acid sensing in the presence of dopamine: electropolymerized poly-pyrrole modified with a reduced graphene oxide/PEDOT:PSS composite.

A highly selective electrochemical sensor based on a molecularly imprinted polymer (MIP) to be developed for uric acid detection in the presence of dopamine as an interference molecule was demonstrated in this study. This non-enzymatic uric acid sensor was developed by electropolymerizing poly-pyrrole onto a composite of electrochemically reduced graphene oxide (ErGO) and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) on a glassy carbon electrode (GCE) to give MIP/ErGO/PEDOT:PSS electrodes. The structural properties, surface morphology, and electrochemical interface of this fabricated uric acid sensor was then characterized using infrared spectroscopy, scanning electron microscopy, atomic force microscopy, and electrochemical impedance spectroscopy. This fabricated non-enzymatic electrochemical sensor (MIP/ErGO/PEDOT:PSS modified GCE) also showed excellent analytical performance at the optimum ratio of monomer/template concentration and optimized electropolymerization conditions, for example in the low concentration range of 0.1-100 µM with a detection limit of 0.05 µM towards uric acid detection in the presence of dopamine. Furthermore, this fabricated uric acid sensor also exhibited good reproducibility and stability for uric acid measurements in the presence of dopamine for 5 consecutive days. In addition, this sensor demonstrates highly selective detection of uric acid in the presence of several interfering species such as dopamine, urea, glucose, magnesium ions, and ascorbic acid. This fabricated uric acid sensor ultimately showed satisfactory uric acid measurement in samples of human urine and is expected to be used in early-stage disease diagnosis.

Optimization of uric acid detection with Au nanorod-decorated graphene oxide (GO/AuNR) using response surface methodology.

A modified glassy carbon electrode (GCE) was developed based on a synthesized graphene oxide (GO) gold nanorod (AuNR) decorated composite (GO/AuNR) for sensitive electrochemical sensing of uric acid (UA). The electrochemical performance of GO/AuNR/GCE for UA detection was investigated employing the differential pulse voltammetry (DPV) technique. Central composite design (CCD) was applied to obtain the optimum composition of the GO and AuNR composite, which provide the highest possible UA oxidation peak current. The optimum composition was obtained at a GO concentration of 5 mg mL-1 and AuNR volume of 10 mL. Under the optimum conditions, GO/AuNR/GCE showed acceptable analytical performance for UA detection with good linearity (concentration range of 10-90 µM) and both a low detection limit (0.4 µM) and quantitation limit (1.0 µM). Furthermore, the proposed sensor exhibits superior stability, reproducibility, and selectivity using ascorbic acid (AA), dopamine (DA), urea, glucose, and magnesium as interferents. Finally, practical use of GO/AuNR/GCE was demonstrated by successfully determining the content of UA in human urine samples with the standard addition approach.

Bifunctional folic-conjugated aspartic-modified Fe3O4 nanocarriers for efficient targeted anticancer drug delivery.

Functionalization of nanocarriers has been considered the most promising way of ensuring an accurate and targeted drug delivery system. This study reports the synthesis of bifunctional folic-conjugated aspartic-modified Fe3O4 nanocarriers with an excellent ability to deliver doxorubicin (DOX), an anticancer drug, into the intercellular matrix. Here, the presence of amine and carboxylate groups enables aspartic acid (AA) to be used as an efficient anchoring molecule for the conjugation of folic acid (FA) (EDC-NHS coupling) and DOX (electrostatic interaction). Based on the results, surface functionalization showed little effect on the physicochemical properties of the nanoparticles but significantly influenced both the loading and release efficiency of DOX. This is primarily caused by the steric hindrance effect due to large and bulky FA molecules. Furthermore, in vitro MTT assay of B16-F1 cell lines revealed that FA conjugation was responsible for a significant increase in the cytotoxicity of DOX-loaded nanocarriers, which was also found to be proportional to AA concentration. This high cytotoxicity resulted from an efficient cellular uptake induced by the over-expressed folate receptors and fast pH triggered DOX release inside the target cell. Here, the lowest IC50 value of DOX-loaded nanocarriers was achieved at 2.814 ± 0.449 µg mL-1. Besides, further investigation also showed that the drug-loaded nanocarriers exhibited less or no toxicity against normal cells.

A facile electrochemical sensor based on a composite of electrochemically reduced graphene oxide and a PEDOT:PSS modified glassy carbon electrode for uric acid detection.

A glassy carbon electrode modified with electrochemically reduced graphene oxide (ErGO) and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) was developed for uric acid determination with dopamine as interference in artificial saliva. The electrochemical performance of the fabricated electrode was studied using both techniques of cyclic voltammetry (CV) and differential pulse voltammetry (DPV), under optimized conditions. Using DPV, the sensor based on ErGO/PEDOT:PSS modified GCE displayed a linear relationship in the concentration ranges of 10-100 µM for uric acid. This uric acid sensor exhibited a high sensitivity with detection limits of 1.08 µM and quantitation limit of 3.61 µM. This sensor also showed good reproducibility for uric acid detection in artificial saliva in 5 consecutive days of measurements. This device was successfully used to analyze uric acid in artificial saliva as well as in a human saliva sample using the standard addition method.

Global diversification and evolutionary history of onchidiid slugs (Gastropoda, Pulmonata).

Many marine species are specialized to specific parts of a habitat. In a mangrove forest, for instance, species may be restricted to the mud surface, the roots and trunks of mangrove trees, or rotting logs, which can be regarded as distinct microhabitats. Shifts to new microhabitats may be an important driver of sympatric speciation. However, the evolutionary history of these shifts is still poorly understood in most groups of marine organisms, because it requires a well-supported phylogeny with relatively complete taxon sampling. Onchidiid slugs are an ideal case study for the evolutionary history of habitat and microhabitat shifts because onchidiid species are specialized to different tidal zones and microhabitats in mangrove forests and rocky shores, and the taxonomy of the family in the Indo-West Pacific has been recently revised in a series of monographs. Here, DNA sequences for onchidiid species from the North and East Pacific, the Caribbean, and the Atlantic are used to reconstruct phylogenetic relationships among Onchidella species, and are combined with new data for Indo-West Pacific species to reconstruct a global phylogeny of the family. The phylogenetic relationships of onchidiid slugs are reconstructed based on three mitochondrial markers (COI, 12S, 16S) and three nuclear markers (28S, ITS2, H3) and nearly complete taxon sampling (all 13 genera and 62 of the 67 species). The highly-supported phylogeny presented here suggests that ancestral onchidiids most likely lived in the rocky intertidal, and that a lineage restricted to the tropical Indo-West Pacific colonized new habitats, including mudflats, mangrove forests, and high-elevation rainforests. Many onchidiid species in the Indo-West Pacific diverged during the Miocene, around the same time that a high diversity of mangrove plants appears in the fossil record, while divergence among Onchidella species occurred earlier, likely beginning in the Eocene. It is demonstrated that ecological specialization to microhabitats underlies the divergence between onchidiid genera, as well as the diversification through sympatric speciation in the genera Wallaconchis and Platevindex. The geographic distributions of onchidiid species also indicate that allopatric speciation played a key role in the diversification of several genera, especially Onchidella and Peronia. The evolutionary history of several morphological traits (penial gland, rectal gland, dorsal eyes, intestinal loops) is examined in relation to habitat and microhabitat evolutionary transitions and suggests that the rectal gland of onchidiids is an adaptation to high intertidal and terrestrial habitats.

Thermosensitive Core-Shell Fe3O4@poly(N-isopropylacrylamide) Nanogels for Enhanced Oil Recovery.

An investigation on the application of thermosensitive core-shell Fe3O4@PNIPAM nanogels in enhanced oil recovery was successfully performed. Here, the unique core-shell architecture was fabricated by conducting the polymerization at the surface of 3-butenoic acid-functionalized Fe3O4 nanoparticles and characterized using X-ray diffraction (XRD), 1H NMR, vibration sample magnetometer (VSM), and high-resolution transmission electron microscopy (HR-TEM). According to the results, this core-shell structure was beneficial for achieving the desired high viscosity and low nanofluid mobility ratio at high temperatures, which is essential for enhanced oil recovery (EOR) application. The results demonstrated that the nanogels exhibited a unique temperature-dependent flow behavior due to the PNIPAM shell's ability to transform from a hydrated to a dehydrated state above its low critical solution temperature (LCST). At such conditions, the nanogels exhibited a significantly low mobility ratio (M = 0.86), resulting in an even displacement front during EOR and leads to higher oil production. Based on the result obtained from sand pack flooding, about 25.75% of an additional secondary oil recovery could be produced when the nanofluid was injected at a temperature of 45 °C. However, a further increase in the flooding temperature could result in a slight reduction in oil recovery due to the precipitation of some of the severely aggregated nanogels at high temperatures.

Modification of boron-doped diamond electrodes with gold-palladium nanoparticles for an oxygen sensor.

Modification of boron-doped diamond (BDD) with gold-palladium nanoparticles (Au@PdNPs) was successfully performed. Prior to the modification, BDD was modified with allylamine to provide active sites for the attachment of nanoparticles, while the synthesis of Au@PdNPs was performed by chemical reduction of a palladium salt solution in a colloidal solution of gold nanoparticles. Characterization using TEM images showed that by controlling the palladium concentration, flower and core-shell shaped Au@PdNPs can be prepared. XPS studies confirmed that the nanoparticles with a flower shape could be attached better on the BDD surface. The Au@PdNPs-modified BDD (Au@PdNPs-BDD) electrodes were then examined for the oxygen reduction reaction in comparison with gold and palladium-based electrodes. One order higher current response was observed at Au@PdNPs-BDD compared to AuNPs-BDD, indicating the contribution of palladium in the oxygen reduction reaction. Good linearity with comparable limits of detection suggested that Au@PdNPs-BDD electrodes are promising for use as oxygen sensors. Furthermore, their application as BOD sensors was demonstrated.

Highly efficient synthesis of biodiesel catalyzed by a cellulose@hematite-zirconia nanocomposite.

The depletion of fossil fuels calls for the development of renewable alternatives such as biodiesel and has inspired much research on catalysts for the production of biodiesel through the esterification of biomass-derived materials. Herein, a green heterogeneous catalyst for highly efficient biodiesel synthesis was fabricated from rice straw-derived cellulose, hematite, and zirconia and was shown to contain porous irregularly shaped α-Fe2O3-ZrO2 composites (average particle size = 42.5 nm) evenly distributed on the nanocellulose surface. The optimal catalyst (nanocellulose:α-Fe2O3-ZrO2 = 2:1, w/w) afforded biodiesel in a yield of 92.50% and with specifications close to those prescribed by international standards. This catalyst could be reused for up to five cycles without a marked activity loss, with the biodiesel yield in the fifth cycle equaling 80.0%. The developed nanocomposite holds great promise for cutting the costs of biodiesel production, as it is derived from biodegradable raw materials and is renewable, non-corrosive, easy to handle, and green. In addition, the large-scale discharge of this catalyst after use does not pose a hazard to the environment.

A Mechanistic Investigation of Sustainable Solvent-Free, Seed-Directed Synthesis of ZSM-5 Zeolites in the Absence of an Organic Structure-Directing Agent.

The solvent-free, seed-directed synthesis using natural precursors has emerged as a sustainable route for the synthesis of zeolite. Albeit the significant progress in the synthesis techniques, the crystallization behaviors of zeolites are somewhat elusive. Herein, we performed a detailed investigation of the crystallization behaviors of ZSM-5 zeolites synthesized through the solvent-free, seed-directed route using rice husk silica as starting materials. The crystallization at 180 °C is completed rapidly in 10 h, with an ultrahigh zeolite yield of at least 95%. Moreover, we evaluated the crystallization kinetics at different temperatures using the nonlinear Avrami equation and found instantaneous nucleation with three-dimensional growth in the studied temperature range, with activation energies for nucleation, transition, and crystal growth of 137, 51, and 51 kJ mol-1, respectively, indicating that nucleation is the rate-determining step. Further investigation of the structural and morphological evolution revealed a preference for secondary nucleation over the seed-growth mechanism. Crystallization proceeds via structural rearrangement within the solid system. We anticipate that our work will provide extensive insights that increase the understanding of zeolite crystallization and expand the highly sustainable production of zeolites.

Systematic revision of the genus Peronia Fleming, 1822 (Gastropoda, Euthyneura, Pulmonata, Onchidiidae).

The genus Peronia Fleming, 1822 includes all the onchidiid slugs with dorsal gills. Its taxonomy is revised for the first time based on a large collection of fresh material from the entire Indo-West Pacific, from South Africa to Hawaii. Nine species are supported by mitochondrial (COI and 16S) and nuclear (ITS2 and 28S) sequences as well as comparative anatomy. All types available were examined and the nomenclatural status of each existing name in the genus is addressed. Of 31 Peronia species-group names available, 27 are regarded as invalid (twenty-one synonyms, sixteen of which are new, five nomina dubia, and one homonym), and four as valid: Peronia peronii (Cuvier, 1804), Peronia verruculata (Cuvier, 1830), Peronia platei (Hoffmann, 1928), and Peronia madagascariensis (Labbé, 1934a). Five new species names are created: P. griffithsi Dayrat & Goulding, sp. nov., P. okinawensis Dayrat & Goulding, sp. nov., P. setoensis Dayrat & Goulding, sp. nov., P. sydneyensis Dayrat & Goulding, sp. nov., and P. willani Dayrat & Goulding, sp. nov. Peronia species are cryptic externally but can be distinguished using internal characters, with the exception of P. platei and P. setoensis. The anatomy of most species is described in detail here for the first time. All the secondary literature is commented on and historical specimens from museum collections were also examined to better establish species distributions. The genus Peronia includes two species that are widespread across the Indo-West Pacific (P. verruculata and P. peronii) as well as endemic species: P. okinawensis and P. setoensis are endemic to Japan, and P. willani is endemic to Northern Territory, Australia. Many new geographical records are provided, as well as a key to the species using morphological traits.

Surface-Functionalized Superparamagnetic Nanoparticles (SPNs) for Enhanced Oil Recovery: Effects of Surface Modifiers and Their Architectures.

Superparamagnetic nanoparticles (SPNs) have been considered as one of the most studied nanomaterials for subsurface applications, including in enhanced oil recovery (EOR), due to their unique physicochemical properties. However, a comprehensive understanding of the effect of surface functionalization on the ability of the nanoparticles to improve secondary and tertiary oil recoveries remains unclear. Therefore, investigations on the application of bare and surface-functionalized SPNs in EOR using a sand pack were carried out in this study. Here, the as-prepared SPNs were functionalized using oleic acid (OA) and polyacrylamide (PAM) to obtain several types of nanostructure architectures such as OA-SPN, core-shell SPN@PAM, and SPN-PAM. Based on the result, it is found that both the viscosity and mobility of the nanofluids were significantly affected by not only the concentration of the nanoparticles but also the type and architecture of the surface modifier, which dictated particle hydrophilicity. According to the sand pack tests, the nanofluid containing SPN-PAM was able to recover as much as 19.28% of additional oil in a relatively low concentration (0.9% w/v). The high oil recovery enhancement was presumably due to the ability of suspended SPN-PAM to act as a mobility control and wettability alteration agent and facilitate the formation of a Pickering emulsion and disjoining pressure.

A new species and new records of Onchidium slugs (Gastropoda, Euthyneura, Pulmonata, Onchidiidae) in South-East Asia.

A new species, Onchidium melakense Dayrat & Goulding, sp. nov., is described, bringing the total to four known species in the genus Onchidium Buchannan, 1800. Onchidium melakense is a rare species with only nine individuals found at three mangrove sites in the Andaman Islands and the Strait of Malacca (western Peninsular Malaysia and eastern Sumatra). The new species is delineated based on mitochondrial (COI and 16S) and nuclear (ITS2 and 28S) DNA sequences as well as comparative anatomy. Each Onchidium species is characterized by a distinct color and can easily be identified in the field, even in the Strait of Malacca where there are three sympatric Onchidium species. An identification key is provided. In addition, Onchidium stuxbergi (Westerlund, 1883) is recorded for the first time from eastern Sumatra, and Onchidium pallidipes Tapparone-Canefri, 1889, of which the type material is described and illustrated here, is regarded as a new junior synonym of O. stuxbergi.

A new genus of air-breathing marine slugs from South-East Asia (Gastropoda, Pulmonata, Onchidiidae).

As part of an ongoing effort to revise the taxonomy of air-breathing, marine, onchidiid slugs, a new genus, Laspionchis Dayrat & Goulding, gen. nov., is described from the mangroves of South-East Asia. It includes two new species, Laspionchis boucheti Dayrat & Goulding, sp. nov., and Laspionchis bourkei Dayrat & Goulding, sp. nov., both distributed from the Malacca Strait to the Philippines and Australia. This study is based on extensive field work in South-East Asia, comparative anatomy, and both mitochondrial (COI and 16S) and nuclear (ITS2 and 28S) DNA sequences. The two new species are found in the same habitat (mud surface in mangrove forests) and are externally cryptic but are distinct anatomically. Both species are also strongly supported by DNA sequences. Three cryptic, least-inclusive, reciprocally-monophyletic units within Laspionchis bourkei are regarded as subspecies: L. bourkei bourkei Dayrat & Goulding, ssp. nov., L. bourkei lateriensis Dayrat & Goulding, ssp. nov., and L. bourkei matangensis Dayrat & Goulding, ssp. nov. The present contribution shows again that species delineation is greatly enhanced by considering comparative anatomy and nuclear DNA sequences in addition to mitochondrial DNA sequences, and that thorough taxonomic revisions are the best and most efficient path to accurate biodiversity knowledge.

Integrative taxonomy of a new and highly-diverse genus of onchidiid slugs from the Coral Triangle (Gastropoda, Pulmonata, Onchidiidae).

A new genus of onchidiid slugs, Wallaconchis Goulding & Dayrat, gen. n., is described, including ten species. Five species were previously described but known only from the type material: Wallaconchis ater (Lesson, 1830), W. graniferum (Semper, 1880), W. nangkauriense (Plate, 1893), W. buetschlii (Stantschinsky, 1907), and W. gracile (Stantschinsky, 1907), all of which were originally classified in Onchidium Buchannan, 1800. Many new records are provided for these five species, which greatly expand their known geographic distributions. Five species are new: Wallaconchis achleitneri Goulding, sp. n., W. comendadori Goulding & Dayrat, sp. n., W. melanesiensis Goulding & Dayrat, sp. n., W. sinanui Goulding & Dayrat, sp. n., and W. uncinus Goulding & Dayrat, sp. n. Nine of the ten Wallaconchis species are found in the Coral Triangle (eastern Indonesia and the Philippines). Sympatry is high, with up to six species found on the island of Bohol (Philippines) and eight species overlapping in northern Sulawesi (Indonesia). Wallaconchis is distinguished from other onchidiids by its bright dorsal colors (red, yellow, orange) but those are extremely variable and not useful for specific identification. Internally, the reproductive system can be used to identify all Wallaconchis species. The copulatory organs of Wallaconchis species are especially diverse compared to other onchidiid genera, and the possible role of reproductive incompatibility in species diversification is discussed. All specimens examined were freshly collected for the purpose of a worldwide revision of the Onchidiidae Rafinesque, 1815. The species are well delineated using DNA sequences and comparative anatomy. Mitochondrial DNA analysis yields thirteen molecular units separated by a large barcode gap, while nuclear DNA yields nine units. By integrating nuclear DNA and mitochondrial DNA with morphology, ten species are recognized. The natural history of each species (e.g., the microhabitat where they are found) is also documented. Nomenclature is addressed thoroughly (the types of all onchidiid species were examined, lectotypes were designated when needed, nomina dubia are discussed). Morphological characters, transitions to new microhabitats, and diversification processes are discussed in the context of a robust molecular phylogeny.

Transient receptor potential vanilloid 4 (TRPV4) expression on the nerve fibers of human dental pulp is upregulated under inflammatory condition.

OBJECTIVE: Transient receptor potential vanilloid 4 (TRPV4) has been considered as a mechano-, thermo- and osmo-receptor. Under inflammatory conditions in dental pulp, teeth can become sensitive upon exposure to a variety of innocuous stimuli. The objective of the present study was to investigate the expression of the TRPV4 channel on nerve fibers in human dental pulp of non-symptomatic and symptomatic teeth associated with inflammatory conditions. DESIGN: Dental pulp from extracted human permanent teeth was processed for fluorescence immunohistochemistry. Ten asymptomatic (normal) and 10 symptomatic (symptoms associated with pulpitis) teeth were used in this study. Nerve fibers were identified by immunostaining for a marker, protein gene product 9.5, and the cells were counterstained with 4',6-diamidino-2-phenylindole. An anti-TRPV4 antibody was used to trace TRPV4 expression. RESULTS: TRPV4 expression was co-localized with the nerve fiber marker. Immunoreactivity for TRPV4 was more intense (p < 0.05) in the nerves of symptomatic teeth than those of normal teeth. The number of co-localization spots was increased significantly (p < 0.05) in the dental pulp of symptomatic teeth compared with that of asymptomatic (normal) teeth. CONCLUSIONS: There is expression of TRPV4 channels on the nerve fibers of human dental pulp. Our findings suggest upregulation of TRPV4 expression under inflammatory conditions in the pulp. The upregulation of TRPV4 channels may be associated with the exaggerated response of dental pulp to innocuous mechanical, thermal and osmotic stimuli under inflammatory conditions.