The New Nematicide Cyclobutrifluram Targets the Mitochondrial Succinate Dehydrogenase Complex in Caenorhabditis elegans.

Today, agriculture around the world is challenged by parasitic nematode infections. Plant-parasitic nematodes (PPNs) can cause significant damage and crop loss and are a threat to food security. For a long time, the management of PPN infection has relied on nematicides that impact not only parasitic nematodes but also other organisms. More recently, new nematicides have been developed that appear to specifically target PPN. Cyclobutrifluram belongs to this new category of nematicides. Using the nematode Caenorhabditis elegans as a model organism, we show here that cyclobutrifluram strongly impacts the survival and fertility rates of the worm by decreasing the number of germ cells. Furthermore, using a genetic approach, we demonstrate that cyclobutrifluram functions by inhibiting the mitochondrial succinate dehydrogenase (SDH) complex. Transcriptomic analysis revealed a strong response to cyclobutrifluram exposure. Among the deregulated genes, we found genes coding for detoxifying proteins, such as cytochrome P450s and UDP-glucuronosyl transferases (UGTs). Overall, our study contributes to the understanding of the molecular mode of action of cyclobutrifluram, to the finding of new approaches against nematicide resistance, and to the discovery of novel nematicides. Furthermore, this study confirms that C. elegans is a suitable model organism to study the mode of action of nematicides.

Cytocompatibility and Antibacterial Properties of Coaxial Electrospun Nanofibers Containing Ciprofloxacin and Indomethacin Drugs.

A coaxial nanofibrous scaffold of poly (ε-caprolactone) and gelatin/cellulose acetate encapsulating anti-inflammatory and antibacterial drugs was co-electrospun for skin tissue regeneration. Indomethacin and ciprofloxacin as model drugs were added to the core and the shell solutions, respectively. The effect of the drugs' presence and crosslinking on the scaffold properties was investigated. TEM images confirmed the core−shell structure of the scaffold. The fiber diameter and the pore size of the scaffold increased after crosslinking. The tensile properties of the scaffold improved after crosslinking. The crosslinked scaffold illustrated a higher rate of swelling, and a lower rate of degradation and drug release compared to the uncrosslinked one. Fitting the release data into the Peppas equation showed that Fickian diffusion was the dominant mechanism of drug release from the scaffolds. The results of biocompatibility evaluations showed no cytotoxicity and suitable adhesion and cell growth on the prepared core−shell structure. The antibacterial activity of the scaffolds was studied against one of the most common pathogens in skin wounds, where the existence of ciprofloxacin could prevent the growth of the Staphylococcus aureus bacteria around the scaffold. The obtained results suggested a new coaxial nanofibrous scaffold as a promising candidate for simultaneous tissue regeneration and controlled drug release.

Morphological and molecular characterization of Filenchus pseudodiscus n. sp. from east Golestan province, north Iran; with an updated phylogeny of Malenchus Andrássy, 1968 (Tylenchomorpha: Tylenchidae).

During a survey in Golestan province, north Iran, two populations belonging to the family Tylenchidae were recovered in association with Quercus sp., and a rotten wood sample of an unidentified forest tree. The first recovered species was mainly characterized by having a disc-like differentiation in the frontal end under the light microscope (LM), proposing it as a tentative member of the genus Discotylenchus. Detailed morphological studies using scanning electron microscope (SEM), however, did not reveal a true disc, but showing the smooth cephalic region, and a narrow annulus behind the cephalic plate. Based upon the cephalic region structure, and by lacking a true disc, the species was identified as a member of the genus Filenchus. This population was further characterized by 555 to 618 µm long females, lateral fields with four incisures, 9 to 10 µm long stylet, spermatheca large, including spheroid sperm, post-vulval uterine sac (PUS) 8 to 12 µm long and gradually tapering to an elongate conoid tail with pointed tip. It was compared with relevant species of Filenchus having four incisures in the lateral fields and similar general morphology. By having a disc-like differentiation in the frontal end under the LM, it was further compared with three similar known species of Discotylenchus. The morphological comparisons with species under two aforementioned genera showed the recovered population belongs to an unknown species, described herein as Filenchus pseudodiscus n. sp. The molecular phylogenetic relationships of the new species using partial small and large subunit ribosomal RNA gene (SSU and LSU D2-D3 rDNA) sequences were reconstructed and discussed. Malenchus gilanensis, the second recovered and studied species was originally established based upon traditional criteria. An updated LSU phylogeny of the genus Malenchus by including M. gilanensis was also presented and its results were discussed.

Description of Longidorella (Saevadorella) caspica n. sp. (Dorylaimida: Nordiidae) from north Iran.

This contribution provides the morphological and molecular identification of a new species of the genus Longidorella (Saevadorella). L. (S.) caspica n. sp., was recovered from the rhizospheric soil of grasses in Mazandaran province in the seashore of the Caspian Sea. It is characterized by females with a length of 788 to 874 µm and a cephalic region with prominent papillae; and separated from the rest of the body by a remarkable constriction, an odontostyle of 32 to 33 µm, vulva at 52.5 to 59.0%, and a tail of 33 to 38 µm with a rounded tip. Males have 32 to 35 µm long spicules of dorylaimoid form and five to seven ventral supplements ending at 48 to 55 µm distance from cloacal pair. The new species was morphologically compared with seven nominal species viz. L. (S.) arenicola, L. (S.) cuspidata, L. (S.) magna, L. (S.) perplexa, L. (S.) saadi, L. (S.) saevae, and L. (S.) tharensis. It was further compared with similar species under the subgenus Longidorella (Enchodorella) viz. L. (E.) deliblatica and L. (E.) murithi. Besides morphological studies, molecular phylogenetic studies using partial sequences of D2 to D3 expansion fragments of 28S rDNA were performed for the new species and its phylogenetic relationships with other species and genera were discussed.

Description of Deladenus brevis n. sp. (Sphaerularioidea: Neotylenchidae) from Iran: a morphological and molecular phylogenetic study.

Deladenus brevis n. sp. is described and illustrated based upon its morphological, morphometric, and molecular characters. The new species is mainly characterized by its small body size (367-454 µm long females and 350, 391 µm long males) and has small lateral vulval flaps. It is further characterized by having six lines in the lateral fields, cephalic region slightly expanded, narrower adjacent to the body, short 6 to 7 µm long stylet with three posteriorly sloped knobs, no post-vulval uterine sac (PUS), conical tail, narrowing to a rounded tip, and rare males with slender tylenchoid spicules and bursa reaching the tail tip. By having six lines in the lateral fields, the new species was compared with five known species of the genus, namely D. apopkaetus, D. cocophilus, D. durus, D. obtusicaudatus, and D. persicus, having comparable number (six or seven) of lines in the lateral fields. It was further compared with D. pakistanensis having lateral vulval flaps. The differences with above-mentioned species are discussed. In molecular phylogenetic analyses using nearly full length small and large subunit ribosomal DNA (SSU and LSU D2-D3 rDNA) and cytochrome c oxidase subunit I (COI mtDNA) gene sequences, D. brevis n. sp. formed clade with species of the genus with different clade support values in Bayesian inference.Deladenus brevis n. sp. is described and illustrated based upon its morphological, morphometric, and molecular characters. The new species is mainly characterized by its small body size (367-454 µm long females and 350, 391 µm long males) and has small lateral vulval flaps. It is further characterized by having six lines in the lateral fields, cephalic region slightly expanded, narrower adjacent to the body, short 6 to 7 µm long stylet with three posteriorly sloped knobs, no post-vulval uterine sac (PUS), conical tail, narrowing to a rounded tip, and rare males with slender tylenchoid spicules and bursa reaching the tail tip. By having six lines in the lateral fields, the new species was compared with five known species of the genus, namely D. apopkaetus, D. cocophilus, D. durus, D. obtusicaudatus, and D. persicus, having comparable number (six or seven) of lines in the lateral fields. It was further compared with D. pakistanensis having lateral vulval flaps. The differences with above-mentioned species are discussed. In molecular phylogenetic analyses using nearly full length small and large subunit ribosomal DNA (SSU and LSU D2-D3 rDNA) and cytochrome c oxidase subunit I (COI mtDNA) gene sequences, D. brevis n. sp. formed clade with species of the genus with different clade support values in Bayesian inference.

Morphological and molecular characterization of Ektaphelenchoides pini () (Aphelenchoididae; Ektaphelenchinae) from Iran, with morphological and taxonomic observations on some species.

Ektaphelenchoides pini, the type species of the genus Ektaphelenchoides, was recovered from wood and bark samples of a dead broadleaf forest tree collected from the forests of Golestan province in north of Iran. The recovered population is mainly characterized by massive wide spicules of males with well-developed condylus marked by indentations at the apex and simple distal tip. It was further characterized by 756 to 947 µm long females having a cephalic region slightly separated from the rest body by a shallow depression, 20 to 23 µm stylet with wide lumen lacking conophore and knobs, excretory pore (E pore) at about one metacorpus length behind it, or 92 to 106 µm from anterior end and hemizonid just posterior to it, vagina anteriorly inclined, post uterine sac (PUS) ca 1.2 times vulval body width long, posterior body region elongate conoid, ending to a filiform tip, no functional rectum, a vestigial anus and common males with dorsally convex tail ending to an elongate spike and two pairs of precloacal (P2) and caudal (P3) large papillae at 5 to 6 µm distance anterior to cloacal opening and 30 to 41% of tail, respectively and lacking the single precloacal papilla (P1). In molecular phylogenetic analyses using small and large subunit ribosomal DNA (SSU, LSU rDNA) sequences, the Iranian population of E. pini fell in a clade including species of three genera Ektaphelenchus, Ektaphelenchoides, and Devibursaphelenchus in SSU, and a clade including species of Ektaphelenchus and Ektaphelenchoides in LSU tree, in close association with an isolate identified as E. pini in the latter phylogeny with high (0.99) Bayesian posterior probability (BPP). The comparisons with the type and French populations, as well as phylogenetic affinities of the species using ribosomal data, are discussed. This is the first report of E. pini from Iran, and its first simultaneous morphological and molecular phylogenetic study. New observations on some species of the genus were also presented and discussed.Ektaphelenchoides pini, the type species of the genus Ektaphelenchoides, was recovered from wood and bark samples of a dead broadleaf forest tree collected from the forests of Golestan province in north of Iran. The recovered population is mainly characterized by massive wide spicules of males with well-developed condylus marked by indentations at the apex and simple distal tip. It was further characterized by 756 to 947 µm long females having a cephalic region slightly separated from the rest body by a shallow depression, 20 to 23 µm stylet with wide lumen lacking conophore and knobs, excretory pore (E pore) at about one metacorpus length behind it, or 92 to 106 µm from anterior end and hemizonid just posterior to it, vagina anteriorly inclined, post uterine sac (PUS) ca 1.2 times vulval body width long, posterior body region elongate conoid, ending to a filiform tip, no functional rectum, a vestigial anus and common males with dorsally convex tail ending to an elongate spike and two pairs of precloacal (P2) and caudal (P3) large papillae at 5 to 6 µm distance anterior to cloacal opening and 30 to 41% of tail, respectively and lacking the single precloacal papilla (P1). In molecular phylogenetic analyses using small and large subunit ribosomal DNA (SSU, LSU rDNA) sequences, the Iranian population of E. pini fell in a clade including species of three genera Ektaphelenchus, Ektaphelenchoides, and Devibursaphelenchus in SSU, and a clade including species of Ektaphelenchus and Ektaphelenchoides in LSU tree, in close association with an isolate identified as E. pini in the latter phylogeny with high (0.99) Bayesian posterior probability (BPP). The comparisons with the type and French populations, as well as phylogenetic affinities of the species using ribosomal data, are discussed. This is the first report of E. pini from Iran, and its first simultaneous morphological and molecular phylogenetic study. New observations on some species of the genus were also presented and discussed.

Nanofibrous scaffolds with biomimetic structure.

This research studies the effect of using a grid-like pattern as a collector on increasing the pore size of the electrospun gelatin/cellulose acetate/elastin scaffolds. The morphological study showed an enlargement in pore size and a decline in fiber diameter in comparison with the scaffold fabricated using conventional flat sheet collectors. The use of the pattern increased the swelling ratio and degradation rate of the scaffold. Investigating the tensile properties of scaffolds revealed that the patterned collector increased the elongation at break up to 145%. In vitro experiments revealed the patterned scaffold as a good substrate for attachment and proliferation of fibroblast cells. Overall, our results indicated that the patterned scaffold of gelatin/cellulose acetate/elastin could provide a better microenvironment for fibroblast cells compared to the conventional scaffolds. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 370-376, 2018.

EPCA2.22: A Silver Lining for Early Diagnosis of Prostate Cancer.

PURPOSE: To investigate whether EPCA-2 (a prostate matrix nuclear protein) can be a more helpful marker in prostate cancer diagnosis. MATERIALS AND METHODS: 176 patients enrolled in this study had abnormal prostate specific antigen (PSA) or digital rectal examination and were candidates for prostate needle biopsy. Blood samples were obtained from each patient prior to biopsy and the samples were frozen for EPCA-2 measurement. Patients diagnosed with cancer were assigned to the case group and those with benign prostate hyperplasia (BPH) were included in the control group. Univariate and multivariable analyses were done to assess the relationship between different independent variables with cancer diagnosis. The diagnostic power of EPCA-2 for cancer was estimated at different levels of PSA according to the ROC curve. RESULTS: The mean(± SD) age of cancer cases was 70.33(± 9.02) years while it was 63.34(± 9.47) years for BPH cases (P < .01). EPCA-2 and PSA were also significantly different between cancer and BPH cases (P < .001). The multivariable logistic regression showed that EPCA-2 has a significant relationship with cancer diagnosis (OR=1.009, P = .021). After controlling other variables following stratification for PSA, it was shown that EPCA-2 and cancer were correlated just when PSA was >10 (P < .001). AUC was 0.694 for cancer prediction by EPCA-2 when PSA was >10 ng/mL. CONCLUSION: EPCA-2 has the power of differentiating BPH from cancer in prostate cancer suspects. This suggests that EPCA-2 can be helpful in diagnosing prostate cancer and can be a preventive test to avoid unnecessary biopsies considering PSA and age of the patient. .

Role of steroid hormone receptors in formation and progression of bladder carcinoma: a case-control study.

PURPOSE: To compare the expression rate of sex steroid hormone receptors of estrogen (ER), progesterone (PR) and androgen (AR) in normal urothelium and urothelial bladder cancer (UBC) and to evaluate the possible associations of these receptors expression with cancer progression and patient's survival. MATERIALS AND METHODS: We evaluated the clinical data and tumor specimens of 120 patients with pathologically confirmed primary UBC with 132 normal healthy controls. Both patients and controls selected from list of subjects who have been referred to Sina Urology clinic, and had a minimum of one year follow-up duration. Data collected from medical cords. For evaluation of expression, immunohistochemistry was performed on paraffin-embedded tissue sections using a monoclonal antibody for androgen, estrogen and progesterone receptors. Presence of at least 10% positive cells defined as positive expression. RESULTS: None of the control subjects showed AR expression, while 22% of the patients were AR-positive. ER/PR expressions were observed in 4.2%/ and 2.5% of the cases and in 2.3% and 1.5% of the controls, respectively. A statistically significant correlation was found between AR expression and tumor stage and grade (P < .001). AR-positive patients showed a significantly poorer prognosis than AR-negative cases (log-rank test, P = .02, hazard ratio = 2.12; 95% confidence interval: 1.36-4.65). CONCLUSION: AR expression was significantly associated with higher grade and poorly differentiated tumors with unfavorable outcome. AR expression test might be useful as a diagnostic tool for determining the malignancy and outcome of UBC patients. 

Phenotypic characterizations and comparison of adult dental stem cells with adipose-derived stem cells.

OBJECTIVES: Mesenchymal stem cells or "multipotent stromal cells" are heterogeneous cell population with self-renewal and multilinage differentiation. The aim of this study was to examine and compare the expression of important stem cell surface markers on two populations of mesenchymal stem cells, one derived from human exfoliated deciduous teeth and the other derived from human adipose tissue. These new stem cells will offer a promising avenue for prevention and reversal of many human diseases such as type 1 diabetes and prevention of liver fibrotic process. METHODS: Mesenchymal stem cells were isolated and cultured from human adipose tissue and dental pulp of human exfoliated deciduous teeth. The cultured cells then were harvested and stained by different fluorescent labeled monoclonal antibodies against surface markers and were analyzed using flow cytometry. RESULTS: Both different cell populations expressed CD44, CD90 and CD13 (stem cell markers) with similar intensity. They did not express hematopoietic markers (CD11b, CD19 and CD34), and lymphocyte or leukocyte antigens CD3, CD7, CD20, CD14, CD45, CCR5 (CD195), CD11b and CD10 on their surfaces. Two different cell types demonstrated different levels of expression in CD56 and CD146. Mesenchymal stem cells from human exfoliated deciduous teeth were positive for CD105 and were negative for CCR3 and CCR4 expression. CONCLUSIONS: Both cell populations derived from adipose tissue and dental pulp showed common phenotypic markers of mesenchymal stem cells. In conclusion, mesenchymal stem cells could be isolated and cultured successfully from dental pulp of human exfoliated deciduous teeth, they are very good candidates for treatment and prevention of human diseases.