Histology and histochemistry of the major salivary glands in the southern white-breasted hedgehog (Erinaceus concolor)

This study aimed to investigate the major salivary glands in the southern white-breasted hedgehog (Erinaceus concolor) histologically and histochemically. Five adult males were included in this study. The results showed that anatomically the shape of the parotid, submandibular and sublingual glands of the Erinaceus concolor was respectively almost pear, elliptical to pyramidal and oval. Histologically, the parotid gland had serous acini and secretory cells showed negative reaction to alcian blue (AB) (pH = 2.5) and negative response to aldehyde fuchsin (AF), methylene blue (MB) and PAS stains. The submandibular gland was a mixed gland of mucous and serous acini. The mucous acini were strongly positive for PAS, AB, MB and AF. However, serous acini were week for PAS and negative against AB, MB and AF stains. The sublingual gland was purely composed of mucous acini. The mucous acini of the sublingual gland were strongly positives for PAS, AB and MB methods. While their reaction to the AF staining was negative. In conclusion, the histological and histochemical observations of the major salivary glands of the southern white-breasted hedgehog (E. concolor) indicated that these glands shown similarities and some special different histochemical features as compared to other mammalian species.     

Preparation of tetrahydroagathic acid: a serum metabolite of isocupressic acid, a cattle abortifacient in ponderosa pine

Isocupressic acid (1) was used to synthetically prepare a mixture of (8S,13R,S)-labda-15,19-dioic acid (tetrahydroagathic acid) (5) via a two-step oxidation procedure followed by hydrogenation of the double bonds at C13 and C8. Reduction of the C8,17 double bond was stereospecific producing only the 8S isomer and confirmed by the nOe interaction between the resulting C17 and C20 methyl groups. The 13R and 13S isomers of 5 were separated and analyzed by HPLC/MS, and (13S)-tetrahydroagathic acid was isolated and identified by comparison to a standard prepared by hydrogenation of naturally occurring (13S)-dihydroagathic acid (4). (13R,S)-dihydroagathic acid was prepared by selective sodium metal-catalyzed hydrogenation of the C13,14 allylic double bond of agathic acid (3). The prepared compounds were then used as standards to confirm the presence of 4 and 5 and their respective 13R and 13S isomers in bovine serum samples. Tetrahydroagathic acid was shown to be the only metabolite detected in serum samples taken from a suspected cattle abortion case submitted for diagnosis; and, thus, 5 could be a valuable diagnostic marker for pine needle-induced abortions.     

The effect of water movement on the transport of dicyandiamide,ammonium and urea in unsaturated soils

A laboratory experiment was conducted to investigate the relative mobility of dicyandiamide (DCD) and jointly applied ammoniacal salts or urea in three different soils of lower Egypt, and to determine the extent to which DCD separates from N-fertilizer in unsaturated soil undergoing leaching. The experimental results suggest that, under conditions of water flow, DCD is readily separated from NH₄⁺ but parts from urea to a far lesser extent. The large difference in mobility between DCD and NH₄⁺ should severely limit the effectiveness of DCD as a nitrification inhibitor in the three soils considered when applied in conjunction with ammoniacal salts. In two out of three cases, the situation is similarly unfavorable in the case of joint DCD and urea application. However, the observation that DCD, in a low CEC sandy loam, moves within the soil solution at a slightly lower rate than urea suggests that joint application with urea would keep at least part of the DCD in contact with the NH₄⁺ ions and, therefore, would preserve some of the effectiveness of DCD under leaching conditions in this soil.     

Simultaneous Analysis of Multidrug Resistance 1(MDR1) C3435T,G2677T/A,and C1236T Genotypes in Hamadan City Population,West of Iran

Background: One of the limitations in the treatment of common diseases such as cancer chemotherapy is development of multidrug resistance (MDR). Polymorphisms could alter the expression level of MDR1 gene, which plays an important role in MDR. In this research, the frequency of C3435T, C1236T, and G2677T/A polymorphisms of MDR1 gene was investigated in a large group of population from Hamadan city to provide a sample data resource. Methods: Peripheral blood (2 ml) was taken, and DNA extraction was carried out. Multiplexed mutagenically separated PCR, which was followed by polyacrylamide gel electrophoresis and silver staining, was applied to detect the mentioned polymorphisms in 935 individuals. Sequencing performed for confirmation of gel electrophoresis resulted in 10 random cases. In total, alleles and genotypes of 933 persons (776 women and 157 men) were determined. Results: The most frequent alleles of the polymorphisms were: 3435T, C1236, and G2677. The most frequent genotypes were: 3435C/T, 1236C/T, and 2677G/A, and their concurrent presence was also found as the most frequent simultaneous genotypes. There was not any meaningful difference among the prevalence of these genotypes in groups of men and women. Conclusion: Our results were close to those of other studies performed in Iran and compared to the other ethnic groups, which showed more similarity to Asian peoples than Europeans. As an aspect of personalized medicine, it could be used by chemotherapists to improve the routine methods of cancer treatment.Key Words: Gene polymorphism, Multi-drug resistance, Neoplasm     

Psammaceratin A: A Cytotoxic Psammaplysin Dimer Featuring an Unprecedented (2Z,3Z)-2,3-Bis(aminomethylene)succinamide Backbone from the Red Sea Sponge Pseudoceratina arabica

Bioassay-guided partition of the extract of the Red Sea sponge Pseudoceratina arabica and HPLC purification of the active fraction gave a psammaplysin dimer, psammaceratin A (1), along with psammaplysin A (2). The dimer comprises two units of psammaplysin A (2) connected via the terminal amines with an unprecedented (2Z,3Z)-2,3-bis(aminomethylene)succinamide moiety, and it represents the first dimer to be identified among the psammaplysin family. Data from 1D- and 2D-NMR and HRMS supported the chemical structures of the compounds. Psammaceratin A (1) and psammaplysin A (2) exhibited significant growth inhibition of HCT 116, HeLa, and MBA-MB-231 cells down to 3.1 μM.     

Subereamolline A as a Potent Breast Cancer Migration, Invasion and Proliferation Inhibitor and Bioactive Dibrominated Alkaloids from the Red Sea Sponge Pseudoceratina arabica

A new collection of several Red Sea sponges was investigated for the discovery of potential breast cancer migration inhibitors. Extracts of the Verongid sponges Pseudoceratina arabica and Suberea mollis were selected. Bioassay-directed fractionation of both sponges, using the wound-healing assay, resulted into the isolation of several new and known brominated alkaloids. Active fractions of the sponge Pseudoceratina arabica afforded five new alkaloids, ceratinines A–E (2–6), together with the known alkaloids moloka’iamine (1), hydroxymoloka’iamine (7) and moloka’iakitamide (8). The active fraction of the sponge Suberea mollis afforded the three known alkaloids subereamolline A (9), aerothionin (10) and homoaerothionin (11). Ceratinine B (3) possesses an unprecedented 5,7-dibrominated dihydroindole moiety with an epoxy ring on the side chain of a fully substituted aromatic moiety. Ceratinines D (5) and E (6) possess a terminal formamide moiety at the ethylamine side chain. Subereamolline A (9) potently inhibited the migration and invasion of the highly metastatic human breast cancer cells MDA-MB-231 at the nanomolar doses. Subereamolline A and related brominated alkaloids are novel scaffolds appropriate for further future use for the control of metastatic breast cancer.     

Bioactive Compounds from the Red Sea Marine Sponge Hyrtios Species

In continuation of our search for drug leads from Red Sea sponges we have investigated the ethyl acetate fraction of the organic extract of the Red Sea sponge Hyrtios species. Bioassay-directed fractionation of the active fraction resulted into the identification of three new alkaloids, hyrtioerectines D–F (1–3). Hyrtioerectines D–F belong to the rare marine alkaloids in which the indole and β-carboline fragments of the molecule are linked through C-3/C-3 of both moieties. The structures of the isolated compounds were established based on different spectroscopic data including UV, IR, 1D and 2D NMR (COSY, HSQC, and HMBC) and high-resolution mass spectral studies. The antimicrobial activity against several pathogens and the free radical scavenging activity of the compounds using DPPH reagent were evaluated. In addition, the growth inhibitory activity of the compounds against three cancer cell lines was also evaluated. Hyrtioerectines D–F (1–3) displayed variable antimicrobial, free radical scavenging and cancer growth inhibition activities. Generally, compounds 1 and 3 were more active than compound 2.     

Development of enzyme-linked immunosorbent assays for isocupressic acid and serum metabolites of isocupressic acid

The consumption of ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), common juniper (Juniperus communis), and Monterey cypress (Cupressus macrocarpa) causes abortions in pregnant cattle. Recent studies have identified isocupressic acid (1) as the primary abortificient compound in these plants. In vitro and in vivo studies using rumen and blood have shown isocupressic acid (1) is rapidly metabolized to agathic acid (3), dihydroagathic acid (4), and tetrahydroagathic acid (5). Rapid and sensitive diagnostic techniques are needed to identify poisoned animals, to study toxicokinetics, and to elucidate the mechanism of isocupressic acid-induced abortion in cattle. In this study, four competitive inhibition enzyme-linked immunosorbent assays for isocupressic acid and its sera metabolites were developed using polyclonal antibodies. One assay is specific to 1, whereas the other three assays show cross-reactivity to 3-5 in addition to 1. The assay specific to 1 had a limit of detection of 44.1 pg. The other assays which demonstrated cross-reactivity to the isocupressic acid blood metabolites also had comparably low limits of detection. One assay was used to follow the absorption and elimination profile of isocupressic acid metabolites in both cow serum and urine after oral dosage of a cow with common juniper.     

Molecular characterization of a Squalene epoxidase gene in dermatophyte pathogen Trichophyton tonsurans

BACKGROUND: Trichophyton tonsurans is one of the dermatophyte fungi which invades the skin and hair of human. Several properties of this fungus have been investigated so far. However a few studies were carried out in the field of molecular biology of this fungus. In the present study, we tried to identify the Squalene epoxidase gene which is related to synthesis of ergosterol in this fungus. METHODS: Pairs of 23 and 24 nucleotides primers were designed from highly conserved regions of the similar genes in other fungi. Mentioned primers were utilized in PCR by using isolated genomic DNA of T. tonsurans whereas the PCR fragments were then sequenced. RESULTS AND CONCLUSION: Nucleotides (n = 558) have been sequenced from this new gene which encodes a polypeptide with 186 amino acids. Sequences comparison in gene data banks (NCBI, NIH) for this part of DNA and its deduced amino acid revealed significant homology with members of the eukaryotic Squalene epoxidase.