Morphological,ultrastructural and phylogenetic analyses of Myxobolus hilarii n. sp. (Myxozoa,Myxosporea), a renal parasite of farmed Brycon hilarii in Brazil

Myxobolus hilarii n. sp. was described, based on morphology, histology, ultrastructure and 18S rDNA sequencing, infecting the kidney of Brycon hilarii (Valenciennes 1850) (Characiformes: Bryconidae) taken from fish farms in the state of São Paulo, Brazil. Thirteen specimens of B. hilarii were examined and 100% had round, white plasmodia in the kidney. The mature myxospores were rounded, measuring 11.5 ± 0.8 (9.8–13.4) μm in length, 11.0 ± 0.7 (9.7–12.4) μm in width and 7.6 ± 1.0 (6.7–9.0) μm in thickness. Polar capsules were elongated and of equal size, with 6.5 ± 0.4 (6.0–7.2) μm in length and 4.0 ± 0.2 (3.6–5.3) μm in width and their polar filaments had 5 to 7 coils. Histological analysis revealed plasmodial development in the renal tubules, causing compression and deformation of adjacent tissues and destruction of renal tubule cells. Ultrastructural analysis showed direct contact between the plasmodial wall and the host tissue and asynchronous plasmodial development. The phylogenetic analysis of South American myxobolids, based on 18S rDNA sequencing, showed the myxosporeans grouping into two main clades. M. hilarii n. sp. appears as sister species of Myxobolus piraputangae.     

Morphology and phylogeny of Henneguya jocu n. sp. (Myxosporea,Myxobolidae), infecting the gills of the marine fish Lutjanus jocu

Henneguya jocu n. sp. (Myxosporea, Myxobolidae) is described from the gill lamellae of the marine teleost fish Lutjanus jocu, with a focus on ultrastructural and molecular features. This myxosporean forms subspherical cysts up to ∼260 μm × 130 μm long, and develops asynchronously. Mature myxospores ellipsoidal with a bifurcated caudal process. Myxospore length 10.9 ± 0.4 μm (n = 50); width, 8.2 ± 0.3 μm (n = 50); and thickness, 2.9 ± 0.5 μm (n = 50). Two equal caudal processes, 34.1 ± 1.0 μm long (n = 50); and total myxospore length, 45.2 ± 1.0 μm (n = 50). Two symmetric valves surround two ellipsoidal polar capsules, 5.0 ± 0.3 × 1.4 ± 0.2 μm (n = 20), each containing an isofilar polar filament forming 4–5 coils along the inner wall of these structures, as well as a binucleated sporoplasm presenting a spherical vacuole and several globular sporoplasmosomes. Both the morphological data and molecular analysis of the SSU rDNA gene identify this parasite as a new species of the genus Henneguya. Maximum Likelihood and Maximum Parsimony analyses further indicate that the parasite clusters within others marine Myxobolidae species, forming a group alongside other Henneguya species described from marine hosts.     

Exogenous and endogenous stages of Eimeria perforans naturally infected domestic rabbit (Oryctolagus cuniculus) in Saudi Arabia: Light microscopic study

Exogenous and endogenous stages of Eimeria perforans naturally infected rabbits in Saudi Arabia were described. The prevalence of infection was 75%. Oocysts were ovoid to elliptical and measured 16 × 10 μm. The four dizoic sporocysts were ovoid and measured 7 × 5 μm. Endogenous stages were restricted to the duodenum. Meronts, microgamonts, macrogamonts and young oocysts were recorded and described.     

Haematological and biochemical parameters and tissue accumulations of cadmium in Oreochromis niloticus exposed to various concentrations of cadmium chloride

Oreochromis niloticus, weighing 36.45 ± 1.12 g were exposed to 10%, 20% and 30% of the LC₅₀ of CdCl₂ which represents treatments (T1)1.68, (T2)3.36 and (T3)5.03 mg/l, respectively, for a period of 10, 20 and 30 days. It was found that, compared to a control group reading of 0.19 ± 0.03 μg/g dry weight, accumulation of Cd in the gills was significantly (p < 0.05) increased in samples ranging between 7.64 ± 0.86 and 61.73 ± 0.82 μg/g dry weight from T1 at 10 days to T3 at 30 days. The accumulation of Cd in the liver, meanwhile, was also observed to significantly increase (p < 0.05) with increasing time and concentrations with results ranging between 3.21 ± 0.12 and 181.61 ± 1.32 compared to the control group results of 0.29 ± 0.04 μg/g dry weight. Although muscles exhibited lower levels of accumulation than the gills and liver they still showed the same pattern of increase compared to the control group, with a significant difference ranging between 0.32 ± 0.02 and 2.16 ± 0.08 compared to the control group results of 0.03 ± 0.001 μg/g dry weight. Also, haematological parameters such as red blood cells (RBCs), haemoglobin (Hb) and haematocrit (Hct) were reduced in fish exposed to Cd at all periods, with significant differences (p < 0.05). Plasma glucose concentration showed a significant increase. Total protein levels of fish showed a significant reduction (p > 0.05) for all exposed treatments. Also, the total lipid level increased significantly as fish were exposed to increasing cadmium concentrations, compared to control fish. Finally, the activities of aspartate aminotransferase (AST IU/l) and alanine aminotransferase (ALT IU/l) showed a significant increase (p < 0.05) with increasing time and concentrations.     

Effect of Zinc nanoparticles on oxidative stress-related genes and antioxidant enzymes activity in the brain of Oreochromis niloticus and Tilapia zillii

This study was carried out to determine the median lethal concentrations (LC₅₀) of Zinc nanoparticles (ZnNPs) on Oreochromis niloticus and Tilapia zillii. The biochemical and molecular potential effects of ZnNPs (500 and 2000 μg L⁻¹) on the antioxidant system in the brain tissue of O. niloticus and T. zillii were investigated. Four hundred fish were used for acute and sub-acute studies. ZnNP LC₅₀ concentrations were investigated in O. niloticus and T. zillii. The effect of 500 and 2000 μg L⁻¹ ZnNPs on brain antioxidants of O. niloticus and T. zillii was investigated. The result indicated that 69 h LC₅₀ was 5.5 ± 0.6 and 5.6 ± 0.4 for O. nilotica and T. zillii, respectively. Fish exposed to 500 μg L⁻¹ ZnNPs showed a significant increase in reduced glutathione (GSH), total glutathione (tGSH) levels, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activity and gene expression. On the contrary, malondialdehyde (MDA) levels significantly decreased. Meanwhile, fish exposed to 2000 μg L⁻¹ ZnNPs showed a significant decrease of GSH, tGSH levels, SOD, CAT, GR, GPx and GST activity and gene expression. On the contrary, MDA levels significantly increased. It was concluded that, the 96 h LC₅₀ of ZnNPs was 5.5 ± 0.6 and 5.6 ± 0.4 for O. nilotica and T. zillii, respectively. ZnNPs in exposure concentrations of 2000 μg/L induced a deleterious effect on the brain antioxidant system of O. nilotica and T. zillii. In contrast, ZnNPs in exposure concentrations of 500 μg L⁻¹ produced an inductive effect on the brain antioxidant system of O. nilotica and T. zillii.     

Cloning,characterization and validation of inosine 5′-monophosphate dehydrogenase of Babesia gibsoni as molecular drug target

The inosine monophosphate dehydrogenase (IMPDH) enzyme has been characterized and validated as a molecular drug target in other apicomplexans but not in the genus Babesia. Subsequently, we cloned and expressed a Babesia gibsoni IMPDH (BgIMPDH) cDNA in Escherichia coli. We also determined the inhibitory effect of mycophenolic acid (MPA) on recombinant BgIMPDH (rBgIMPDH) activity and the Babesia-growths in vitro. The translated BgIMPDH peptide contained thirteen amino acid residues responsible for substrate and cofactor binding in its catalytic domain with Gly374 in BgIMPDH being replaced by Ser388 in mammalian IMPDH. The native BgIMPDH enzyme in the parasite was approximately 54-kDa a mass similar to His-tag rBgIMPDH protein. The K_m values of the rBgIMPDH were 8.18 ± 0.878 (mean ± standard error of the mean) μM and 360.80 ± 43.41 μM for IMP and NAD⁺, respectively. MPA inhibited the rBgIMPDH activity yielding a K_i value of 20.93 ± 1.83 μM with respect to NAD⁺. For Babesia growths, the IC₅₀s were 0.95 ± 0.21 and 2.88 ± 0.49 μM for B. gibsoni and B. bovis, respectively. Therefore, our results suggest that MPA may inhibit the replication of Babesia parasites by targeting IMPDH enzyme of the purine pathway.     

Synthesis and biological evaluation of indole derivatives as α-amylase inhibitor

A series of twenty indole hydrazone analogs (1–21) were synthesized, characterized by different spectroscopic techniques such as ¹H NMR and EI-MS, and screened for α-amylase inhibitory activity. All analogs showed a variable degree of α-amylase inhibition with IC₅₀ values ranging between 1.66 and 2.65 μM. Nine compounds that are 1 (2.23 ± 0.01 μM), 8 (2.44 ± 0.12 μM), 10 (1.92 ± 0.12 μM), 12 (2.49 ± 0.17 μM), 13 (1.66 ± 0.09 μM), 17 (2.25 ± 0.1 μM), 18 (1.87 ± 0.25 μM), 20 (1.83 ± 0.63 μM), and 19 (1.97 ± 0.02 μM) showed potent α-amylase inhibition when compared with the standard acarbose (1.05 ± 0.29 μM). Other analogs showed good to moderate α-amylase inhibition. The structure activity relationship is mainly focusing on difference of substituents on phenyl part. Molecular docking studies were carried out to understand the binding interaction of the most active compounds.     

Biomorphometric characteristics of different types of sensilla detected on the antenna of Helicoverpa armigera by scanning electron microscopy

A morphometric study on H. armigera antenna showed four styles of sensilla, i.e., styloconica, chaetica, coeloconica, and trichodea, and their numbers were estimated. Sensilla trichodea detect inter and intraspecific communication signals and was the most numerous. They were divided into three types: type I, the longest, with a length of 34.04 ± 3.16 μm and about 2.16 to 2.42 μm in diameter at its base; 2) type II, intermediate, with a length of 22.58 ± 0.77 μm and basal diameter of 1.8–2.52 μm; 3) type III, the shortest sensilla trichodea, with a length of 7.62 ± 0.4 μm and a range in diameter similar to that of type II. The length of the female sensilla trichodea was longer than that of the male. The total number of sensilla trichodea was estimated to be 7520 on the antenna of the female, and 6831 on the male antenna. The lengths of the sensilla trichodea type I and type III were significantly different on male (t = 4.6881, P = 0.0034) and female antenna (t = 18.9852, P = 0.0001). An estimation of the predicted surface area of the most numerous type I on sampled segments between the 12th and 20th segments from a female of H. armigera showed a surface area of 5 × 10³ μm² and a sensillar density of 38 sensilla/10³ μm². The fraction of sensilla-occupied surface area was 0.4 μm².     

Eye lens dose correlations with personal dose equivalent and patient exposure in paediatric interventional cardiology performed with a fluoroscopic biplane system

PurposeTo analyse the correlations between the eye lens dose estimates performed with dosimeters placed next to the eyes of paediatric interventional cardiologists working with a biplane system, the personal dose equivalent measured on the thorax and the patient dose.MethodsThe eye lens dose was estimated in terms of H_p(0.07) on a monthly basis, placing optically stimulated luminescence dosimeters (OSLDs) on goggles. The H_p(0.07) personal dose equivalent was measured over aprons with whole-body OSLDs. Data on patient dose as recorded by the kerma-area product (P_KA) were collected using an automatic dose management system. The 2 paediatric cardiologists working in the facility were involved in the study, and 222 interventions in a 1-year period were evaluated. The ceiling-suspended screen was often disregarded during interventions.ResultsThe annual eye lens doses estimated on goggles were 4.13 ± 0.93 and 4.98 ± 1.28 mSv. Over the aprons, the doses obtained were 10.83 ± 0.99 and 11.97 ± 1.44 mSv. The correlation between the goggles and the apron dose was R² = 0.89, with a ratio of 0.38. The correlation with the patient dose was R² = 0.40, with a ratio of 1.79 μSv Gy⁻¹ cm⁻². The dose per procedure obtained over the aprons was 102 ± 16 μSv, and on goggles 40 ± 9 μSv. The eye lens dose normalized to P_KA was 2.21 ± 0.58 μSv Gy⁻¹ cm⁻².ConclusionsMeasurements of personal dose equivalent over the paediatric cardiologist’s apron are useful to estimate eye lens dose levels if no radiation protection devices are typically used.     

Effects of gender and age on median and ulnar nerve sensory responses over ring finger

ObjectiveTo analyze the effect of gender on median nerve (MN) and ulnar nerve (UN) sensorial responses over ring finger (RF).Materials and methodsResults of individuals admitted to our ENMG laboratory between June 2011 and March 2012 for nerve conduction studies (NCSs) were retrospectively analyzed. Sensory NCSs were performed by standard antidromic technique.ResultsTotally, 112 normal recordings belong to 100 patients were included. Mean antidromic sensory conduction velocity of MNs (wrist-to-second finger) or UNs (wrist-to-fifth finger) was not different between two genders. Mean sensory nerve action potential (SNAP) amplitude of MN from second finger was also not different between two genders. However, mean SNAP amplitude of UN from fifth finger was higher in females. In RF’s sensorial response studies; mean peak latency of MN was similar between females and males (3.05 ± 0.25 ms vs. 3.14 ± 0.29 ms, p = 0.111), whereas one of UN was shorter in females (2.86 ± 0.22 ms vs. 3.04 ± 0.31 ms, p = 0.001). MN to UN latency difference to RF was greater in females than males (0.19 ± 0.15 ms vs. 0.10 ± 0.16 ms, p = 0.007). Mean SNAP amplitude of MN and UN were both higher in females than males (17.9 ± 7.1 μV vs. 14.1 ± 5.5 μV, p = 0.011 and 18.5 ± 8.0 μV vs. 12.9 ± 6.1 μV, p = 0.0009, respectively). All data of NCSs were re-analyzed after adjustment for age, and obtained findings regarding effect of aging are also included.ConclusionGender has a prominent effect on RF’s sensorial responses. Normative values regarding them should be prepared with adjustment for gender.     

Derivatives of schisandrin with increased inhibitory potential on prostaglandin E2 and leukotriene B4 formation in vitro

Four derivatives of schisandrin, a major dibenzo[a,c]cyclooctadiene lignan of Schisandra chinensis (Turcz.) Baillon were synthesized and structurally characterized by means of NMR and mass spectroscopy. Furthermore, axial chirality of the biphenyl system was determined by comparison of calculated with measured circular dichroism (CD) spectra. Three of the obtained derivatives showed a ring contraction during chemical modification. While the original lignans were inactive on the performed bioassays, the compounds which showed the cycloheptadiene skeleton revealed remarkable activities. For the inhibition of LTB₄ production the IC₅₀ values of aR-6,7-dihydro-6-(1′-hydroxyethyl)-3,9-dimethoxy-6-methyl-5H-dibenzo[a,c]cycloheptene-1,2,10,11-tetraol (6) and aR-6-(1′-iodoethyl)-1,2,3,9,10,11-hexamethoxy-6-methyl-5H-dibenzo[a,c]cycloheptene (8) were 4.2 ± 0.3 μM and 4.5 ± 0.2 μM, respectively. aR-6,7-Dihydro-6-(1′-hydroxyethyl)-6-methyl-5H-dibenzo[a,c]cycloheptene-1,2,3,9,10,11-hexaol (5) revealed dual inhibition on COX-2 (IC₅₀ 32.1 ± 2.5 μM) and on LTB₄ production (37.3 ± 5.5% inhibition at 50 μM).     

Oxidative stress and DNA damage in relation to transition metals overload in Abu-Qir Bay,Egypt

The aim of the present study is to evaluate the transition metals overload in Abu-Qir Bay in Egypt, as compared to a less polluted area (reference area) through some biomarkers of oxidative stress. Catalase enzyme activity, malondialdehyde (MDA) concentration and DNA damage (number of apurinic/apyrimidinic sites) were the tested biomarkers. The levels of iron and copper in Mugil cephalus liver tissues were significantly higher in samples from the polluted area as compared to the reference area: Fe: 407 ± 38 vs. 216 ± 21 μg/g wet wt; p = 0.008, Cu: 54 ± 6 vs. 17.7 ± 4 μg/g wet wt; p = 0.0001. This could account for the observed increase in MDA concentration (15.7 ± 5.7 vs. 2.5 ± 0.5 U/g; p = 0.035), and the elevated number of AP sites (13.9 ± 2.6 vs. 0.37 ± 0.2 AP site/1 × 10⁵ bp; p = 0.0001). Similarly, the activity of catalase enzyme responsible for the cellular defense was significantly high (58.3 ± 12.2 vs. 28.4 ± 4.0 U/mg; p = 0.032). The present data indicated a clear relationship between the pollution degree of the above marine environment and both biochemical and molecular responses of the piscine system.     

Analysis of the germination rate,mitotic index and karyotype of Chromolaena barranquillensis (Hieron.) R.M. King & H. Rob.—Asteraceae

Chromolaena barranquillensis (Asteraceae) is an endemic plant of northern Colombia that has garnered economic and medicinal interest, because species from the genus Chromolaena have shown diverse biological activities. This study describes, for the first time, the karyotype, germination and mitotic indices of C. barranquillensis (Hieron.) R.M. King & H. Rob. The germination index was between 34% and 56% with an average germination rate of 1.2 ± 0.4 seeds/day. The mitotic index analysis allowed to determine the cell cycle time (4 h, 10 min) and the mitotic hours (3:00–8:00 h and 17:00 h). The mitosis time was 49 min, equivalent to ~ 20% of the cell cycle. Karyotype analysis showed that C. barranquillensis is a hexaploid species with a chromosomal formula 2n = 6x = 60 = 48 m + 12 sm, and the average chromosomal lengths were 1.7 ± 0.1 μm to 0.9 ± 0.3 μm. The Stebbins asymmetry index was 2B, and the total form percentage was ~ 41%. These results uncover differences between C. barranquillensis and Chromolaena odorata, one of the most abundant species found in the world and the most closely related species to C. barranquillensis.     

Coumarin-thiazole and -oxadiazole derivatives: Synthesis,bioactivity and docking studies for aldose/aldehyde reductase inhibitors

In continuation of our previous efforts directed towards the development of potent and selective inhibitors of aldose reductase (ALR2), and to control the diabetes mellitus (DM), a chronic metabolic disease, we synthesized novel coumarin-thiazole 6(a–o) and coumarin-oxadiazole 11(a–h) hybrids and screened for their inhibitory activity against aldose reductase (ALR2), for the selectivity against aldehyde reductase (ALR1). Compounds were also screened against ALR1. Among the newly designed compounds, 6c, 11d, and 11g were selective inhibitors of ALR2. Whereas, (E)-3-(2-(2-(2-bromobenzylidene)hydrazinyl)thiazol-4-yl)-2H-chromen-2-one 6c yielded the lowest IC₅₀ value of 0.16 ± 0.06 μM for ALR2. Moreover, compounds (E)-3-(2-(2-benzylidenehydrazinyl)thiazol-4-yl)-2H-chromen-2-one (6a; IC₅₀ = 2.94 ± 1.23 μM for ARL1 and 0.12 ± 0.05 μM for ARL2) and (E)-3-(2-(2-(1-(4-bromophenyl)ethylidene)hydrazinyl)thiazol-4-yl)-2H-chromen-2-one (6e; IC₅₀ = 1.71 ± 0.01 μM for ARL1 and 0.11 ± 0.001 μM for ARL2) were confirmed as dual inhibitors. Furthermore, compounds 6i, 6k, 6m, and 11b were found to be selective inhibitors for ALR1, among which (E)-3-(2-(2-((2-amino-4-chlorophenyl)(phenyl)methylene)hydrazinyl)thiazol-4-yl)-2H-chromen-2-one (6m) was most potent (IC₅₀ = 0.459 ± 0.001 μM). Docking studies performed using X-ray structures of ALR1 and ALR2 with the given synthesized inhibitors showed that coumarinyl thiazole series lacks the carboxylate function that could interact with the anionic binding site being a common ALR1/ALR2 inhibitors trait. Molecular docking study with dual inhibitor 6e also suggested plausible binding modes for the ALR1 and ALR2 enzymes. Hence, the results of this study revealed that coumarinyl thiazole and oxadiazole derivatives could act as potential ALR1/ALR2 inhibitors.     

Synthesis and activity of novel tetrazole compounds and their pyrazole-4-carbonitrile precursors against Leishmania spp

A new series of 5-(1-aryl-3-methyl-1H-pyrazol-4-yl)-1H-tetrazole derivatives (4a–m) and their precursor 1-aryl-3-methyl-1H-pyrazole-4-carbonitriles (3a–m) were synthesized and evaluated as antileishmanials against Leishmania braziliensis and Leishmania amazonensis promastigotes in vitro. In parallel, the cytotoxicity of these compounds was evaluated on the RAW 264.7 cell line. The results showed that among the assayed compounds the substituted 3-chlorophenyl (4a) (IC₅₀/24 h = 15 ± 0.14 μM) and 3,4-dichlorophenyl tetrazoles (4d) (IC₅₀/24 h = 26 ± 0.09 μM) were the most potent against L. braziliensis promastigotes, as compared the reference drug pentamidine, which presented IC₅₀ = 13 ± 0.04 μM. In addition, 4a and 4d derivatives were less cytotoxic than pentamidine. However, these tetrazole derivatives (4) and pyrazole-4-carbonitriles precursors (3) differ against each of the tested species and were more effective against L.braziliensis than on L. amazonensis.     

Changes in the size distribution of goat steroidogenic luteal cells during pregnancy

Experiments were conducted to investigate the size distribution of goat steroidogenic luteal cells throughout pregnancy. Corpora lutea were collected from very early (<6 weeks), early (6–8 weeks), middle (9–14 weeks) or late (15–18 weeks) stages of pregnancy. Luteal tissue was dissociated into single-cell suspension by enzyme treatments. Cells were stained for 3β-hydroxysteroid dehydrogenase (3β-HSD) activity, a marker for steroidogenic cells. The steroidogenic cells covered a wide spectrum of size ranging from 5 to 45 μm in diameter. There was a significant increase in mean cell diameter (P>0.01) as pregnancy progressed. Mean diameter of 3β-HSD positive cells increased from 14.73±0.35 μm in the corpus luteum of very early pregnancy to 24.20±0.45 μm in the corpus luteum of late pregnancy. The ratio of large (>20 μm in diameter) to small (5–20 μm in diameter) luteal cells was 0.28:1.0 in very early pregnancy, with the 7.5–15 μm cell size class being dominant. However, the ratio of large-to-small luteal cells was increased to 1.77:1.0 μm as pregnancy advanced and 25–35 μm cell sizes became predominant. It is likely that small luteal cells could develop into large cells as pregnancy progresses. Development of pregnancy is also associated with an increase in size of steroidogenic luteal cells.     

Design,synthesis, and docking studies of phenylpicolinamide derivatives bearing 1H-pyrrolo[2,3-b]pyridine moiety as c-Met inhibitors

Four series of phenylpicolinamide derivatives bearing 1H-pyrrolo[2,3-b]pyridine moiety (12a–e, 13a–f, 14a–f and 15a–i) were designed, synthesized and evaluated for the IC₅₀ values against three cancer cell lines (A549, PC-3 and MCF-7) and c-Met kinase. Five selected compounds (13b, 15b, 15d, 15e and 15f) were further evaluated for the activity against HepG2 and Hela cell lines. Eighteen of the compounds showed excellent cytotoxicity activity and selectivity with the IC₅₀ valuables in single-digit μM to nanomole range. Seven of them are equal to more active than positive control Foretinib against one or more cell lines. The most promising compound 15f showed superior activity to Foretinib, with the IC₅₀ values of 1.04 ± 0.11 μM, 0.02 ± 0.01 μM and 9.11 ± 0.55 μM against A549, PC-3 and MCF-7 cell lines, which were 0.62 to 19.5 times more active than Foretinib (IC₅₀ values: 0.64 ± 0.26 μM, 0.39 ± 0.11 μM, 9.47 ± 0.22 μM), respectively. Structure–activity relationships (SARs) and docking studies indicated that replacement of quinoline nucleus of the previous active compounds with 1H-pyrrolo[2,3-b]pyridine moiety maintained even improved the potent cytotoxic activity. The results suggested that the introduction of fluoro atoms to the aminophenoxy part of target compounds or the phenyl group of pyrimidine substituted on C-4 position was benefit for the activity.     

Glomus drummondii and G. walkeri,two new species of arbuscular mycorrhizal fungi (Glomeromycota)

Two new ectocarpic arbuscular mycorrhizal fungal species, Glomus drummondii and G. walkeri (Glomeromycota), found in maritime sand dunes of northern Poland and those adjacent to the Mediterranean Sea are described and illustrated. Mature spores of G. drummondii are pastel yellow to maize yellow, globose to subglobose, (58–)71(–85) μm diam, or ovoid, 50–80 × 63–98 μm. Their wall consists of three layers: an evanescent, hyaline, short-lived outermost layer, a laminate, smooth, pastel yellow to maize yellow middle layer, and a flexible, smooth, hyaline innermost layer. Spores of G. walkeri are white to pale yellow, globose to subglobose, (55–)81(–95) μm diam, or ovoid, 60–90 × 75–115 μm, and have a spore wall composed of three layers: a semi-permanent, hyaline outermost layer, a laminate, smooth, white to pale yellow middle layer, and a flexible, smooth, hyaline innermost layer. In Melzer's reagent, only the inner- and outermost layers stain reddish white to greyish rose in G. drummondii and G. walkeri, respectively. Both species form vesicular–arbuscular mycorrhizae in one-species cultures with Plantago lanceolata as the host plant. Phylogenetic analyses of the ITS and parts of the LSU of the nrDNA of spores placed both species in Glomus Group B sensu Schüßler et al. [Schüßler A, Schwarzott D, Walker C, 2001. A new fungal phylum, the Glomeromycota: phylogeny and evolution. Mycolological Research 105: 1413-1421.]     

Synthesis of pyrimidine-2,4,6-trione derivatives: Anti-oxidant,anti-cancer, α-glucosidase, β-glucuronidase inhibition and their molecular docking studies

This paper describes a facile protocol, efficient, and environmentally benign for the synthesis a series of barbiturate acid substituted at C5 position 3a–o. The desired compounds subjected in vitro for different set of bioassays including against anti-oxidant (DPPH and super oxide scavenger assays), anti-cancer, α-glucosidase and β-glucuronidase inhibitions. Compound 3m (IC₅₀ = 22.9 ± 0.5 μM) found to be potent α-glucosidase enzyme inhibitors and showed more activity than standard acarbose (IC₅₀ = 841 ± 1.73 μM). Compound 3f (IC₅₀ = 86.9 ± 4.33 μM) found to be moderate β-Glucuronidase enzyme inhibitors and showed activity comparatively less than the standard d-saccharic acid 1,4-lactone (IC₅₀ = 45.75 ± 2.16 μM). Furthermore, in sillico investigation was carried out to investigate bonding mode of barbiturate acid derivatives.     

Different effects of ATP on the contractile activity of mice diaphragmatic and skeletal muscles

Apart from acetyl-choline (Ach), adenosine-5′-trisphosphate (ATP) is thought to play a role in neuromuscular function, however little information is available on its cellular physiology. As such, effects of ATP and adenosine on contractility of mice diaphragmatic and skeletal muscles (m. extensor digitorum longa—MEDL) have been investigated in in vitro experiments. Application of carbacholine (CCh) in vitro in different concentrations led to pronounced muscle contractions, varying from 9.15 ± 4.76 to 513.13 ± 15.4 mg and from 44.65 ± 5.01 to 101.46 ± 9.11 mg for diaphragm and MEDL, respectively. Two hundred micromolars of CCh in both muscles caused the contraction with the 65% (diaphragm) to 75% (MEDL) of maximal contraction force—this concentration was thus used in further experiments. It was found that application of ATP (100 μM) increased the force of diaphragmatic contraction caused by CCh (200 μM) from 335.2 ± 51.4 mg (n = 21) in controls to 426.5 ± 47.8 mg (n = 10; P < 0.05), but decreased the contractions of MEDL of CCh from 76.6 ± 6.5 mg (n = 26) in control to 40.2 ± 9.0 mg (n = 8; P < 0.05). Application of adenosine (100 μM) had no effect on CCh-induced contractions of these muscles.Resting membrane potential (MP) measurements using sharp electrodes were done at 10, 20 and 30 min after the application of ATP and adenosine. Diaphragm showed depolarization from 75 ± 0.6 down to 63.2 ± 1.05, 57.2 ± 0.96 and 53.6 ± 1.1 mV after 10, 20 and 30 min of exposition, respectively (20 fibers from 4 muscles each, P < 0.05 in all three cases). Adenosine showed no effect on diaphragmatic MP. Both agents were ineffective in case of MEDL.The effects of ATP in both tissues were abolished by suramin (100 μM), a P2-receptor antagonist, and chelerythrin (50 μM), a specific protein-kinase C (PKC) inhibitor, but were not affected by 1H-[1,2,4]-oxadiazolo-[4,3-α]-quinoxalin-1-one (ODQ, 1 μM), a guanylyl-cyclase inhibitor, or by adenosine-3,5-monophosphothioate (Rp-cAMP, 1 μM), a protein-kinase A (PKA) inhibitor.Besides the action on contractile activity, ATP (100 μM) led to a significant (P < 0.001) depolarization of diaphragm muscle fibers from 74.5 ± 2.3 down to 64 ± 2.1, 58.2 ± 2.2 and 54.3 ± 2.4 mV after 10, 20 and 30 min of incubation, respectively. Incubation of MEDL with the same ATP concentration showed no significant change of MP.Denervation of the muscles for 28 days led to a decrease of CCh-induced contractions of diaphragm down to 171.1 ± 34.5 mg (n = 11, P < 0.05), but increased the contractile force of MEDL up to 723.9 ± 82.3 mg (n = 9, P < 0.01). Application of ATP elevated the contractility of denervated diaphragm caused by CCh up to normal values (311.1 ± 79.7 mg, n = 6, P > 0.05 versus control), but did not significantly affect of contractility of MEDL, which became 848.1 ± 62.7 mg (n = 6).These results show that the effects of ATP on both diaphragmatic and skeletal muscles are mediated through P2Y receptors coupled to chelerytrin-sensitive protein-kinase C.